RESUMO
AIMS: The importance of bacterioferritin in the virulence and pathogenicity of the genus Mycobacterium is still unclear. The aim of this study was to analyse if the expression of a recombinant bacterioferritin from M. tuberculosis (Mtb) by Mycma could improve the capacity of this bacillus to resist the host defence mechanisms. METHODS AND RESULTS: Recombinant Mycma, expressing bacterioferritin (Rv1876) from Mtb, was developed by transformation with pMIP12_Rv1876. To determine bacterioferritin influence on Mycma physiology and virulence, the mycobacteria growth was analysed in vitro and in vivo. It was observed that the expression of bacterioferritin improved the growth rate of recombinant Mycma_BfrA under iron excess and oxidative stress, as compared to the wild type. Furthermore, in the murine model of infection, it was observed that Mycma_BfrA-infected mice had higher bacillary load and a more pronounced lesion in the lungs when compared with the wild type. CONCLUSION: This study showed that bacterioferritin confers additional resistance to stress conditions, resulting in increased pathogenicity of Mycma during mice infection. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new insights about the importance of bacterioferritin in the virulence and pathogenicity of the Mycobacterium genus.
Assuntos
Proteínas de Bactérias/metabolismo , Grupo dos Citocromos b/metabolismo , Ferritinas/metabolismo , Mycobacterium abscessus/fisiologia , Mycobacterium abscessus/patogenicidade , Animais , Carga Bacteriana , Proteínas de Bactérias/genética , Grupo dos Citocromos b/genética , Ferritinas/genética , Camundongos , Infecções por Mycobacterium não Tuberculosas/microbiologia , Infecções por Mycobacterium não Tuberculosas/patologia , Mycobacterium abscessus/genética , Mycobacterium abscessus/crescimento & desenvolvimento , Mycobacterium tuberculosis/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Estresse Fisiológico , VirulênciaRESUMO
Male obesity is associated with decreased testosterone levels but the pathophysiological mechanisms behind this association are not completely understood. This study aimed to investigate the impact of hyperglycaemia/insulin resistance and sex hormone-binding globulin (SHBG) levels on testosterone levels in a population of obese men. We investigated the impact of several clinical, anthropometric and analytic measures on testosterone levels in 150 obese males. Testosterone deficiency was present in 52.0% of the enrolled patients. This percentage dropped to 17.6% when only calculated free testosterone (FT) was accounted, as SHBG levels were correlated negatively with body mass index (r = -.20; p < .05). Older age (p < .05) and higher homoeostasis model assessment of insulin resistance (HOMA-IR) (p < .01) and lower SHBG levels (p < .05) were independently correlated with lower FT. Weight and fasting plasma glucose lost their statistical significance after multivariate adjustment. Patients with type 2 diabetes mellitus and pre-diabetes had lower FT than those with normal glucose tolerance (p < .05 and p < .01 respectively). Insulin resistance, and not hyperglycaemia and weight per se, seems to be the main determinant of low testosterone levels in obese males. Low SHBG levels are correlated with low FT even after HOMA-IR adjustment. This suggests that SHBG can be associated with testosterone deficiency beyond the influence of insulin resistance unlike previously reported.
Assuntos
Resistência à Insulina , Obesidade/complicações , Globulina de Ligação a Hormônio Sexual/análise , Testosterona/deficiência , Tecido Adiposo , Adulto , Glicemia , Índice de Massa Corporal , Estudos Transversais , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/metabolismo , Estudos Retrospectivos , Testosterona/sangueAssuntos
Anemia Hemolítica , Hemoglobinas Anormais , Mutação , Adulto , Anemia Hemolítica/sangue , Anemia Hemolítica/genética , Anemia Hemolítica/patologia , Anemia Hemolítica/terapia , Brasil , Feminino , Hemoglobinas Anormais/química , Hemoglobinas Anormais/genética , Hemoglobinas Anormais/metabolismo , HumanosRESUMO
Schistosoma mansoni is a blood fluke parasite responsible for schistosomiasis. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. In this study, we cloned, expressed and purified SmTSP-2 fused to the N- and C-terminal halves of Sm29 and tested these chimeras as vaccine candidates using an adjuvant approved to be used in humans. The results demonstrated that vaccination with SmTSP-2 fused to N- or C-terminus of Sm29-induced reduction in worm burden and liver pathology when compared to control animals. Additionally, we detected high levels of mouse-specific IgG, IgG1 and IgG2a against both chimeras and significant amounts of IFN-γ and TNF-α and no IL-4. Finally, studies with sera from patients resistant to infection and living in schistosomiasis endemic areas revealed high levels of specific IgG to both chimeras when compared to healthy individuals. In conclusion, SmTSP-2/Sm29 chimeras tested here induced partial protection against infection and might be a potential vaccine candidate.
Assuntos
Antígenos de Bactérias/imunologia , Antígenos de Helmintos/imunologia , Proteínas de Bactérias/imunologia , Proteínas de Helminto/imunologia , Glicoproteínas de Membrana/imunologia , Schistosoma mansoni , Esquistossomose mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Tetraspaninas/imunologia , Vacinas/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Bactérias/administração & dosagem , Antígenos de Helmintos/administração & dosagem , Proteínas de Bactérias/administração & dosagem , Ilhas de CpG , Citocinas/sangue , Feminino , Proteínas de Helminto/administração & dosagem , Humanos , Imunoglobulina G/sangue , Fígado/patologia , Glicoproteínas de Membrana/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Oligodesoxirribonucleotídeos/administração & dosagem , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/imunologia , Tetraspaninas/administração & dosagem , Vacinas/administração & dosagemRESUMO
OBJECTIVES: Trauma to the oral tissues can be caused by fellatio. Few cases are reported in the literature. METHODS: A case of oral palate lesions is presented and discussed. RESULTS: The patient developed a large band of petechial haemorrhage extending across the soft palate following the practice of fellatio. The diagnosis was made through an interview with the patient, which disclosed a probable case for fellatio as the cause of the palatal spots. At the follow-up visit 14 days later, the lesions disappeared. CONCLUSIONS: Oral sex activity has increased over the last decades. The dental care professional should be aware that lesions of the palate may result from sexual behaviour. With this possibility in mind, those working in the area of head and neck medicine should consider fellatio as an addition to the differential aetiology of intraoral petechiae.
Assuntos
Mucosa Bucal/lesões , Palato Mole/lesões , Púrpura/etiologia , Comportamento Sexual , Diagnóstico Diferencial , Feminino , Seguimentos , Humanos , Fotografação , Púrpura/diagnóstico , Adulto JovemRESUMO
OBJECTIVE: Mancozeb is one of the most widely used Ethylenebisdithiocarbamates fungicides in Brazil. A pilot experimental model was created to evaluate its potential hepatotoxic effect. MATERIALS AND METHODS: An experimental study was performed with 27 male Wistar rats (3 groups). The Control Group received a saline solution, while Intervention Groups I and II received 250 mg/kg and 500 mg/kg of Mancozeb respectively, once a week, for 12 weeks. Anthropometric measurements were carried out, and the marker of biological exposure in urine was dosed. Biochemical tests, evaluation micronucleus count, comet and oxidative stress markers assay, and histological assessment of the liver were also performed. RESULTS: The hepatotoxic effect of Mancozeb was confirmed by anthropometric measurements, genotoxicity, and oxidative stress. Statistically significant results were found when the exposed groups were compared to the control group. CONCLUSIONS: These results were supported by inflammatory infiltration and balloonization in the treated groups. The experimental model effectively demonstrated the deleterious effect of Mancozeb on the liver.
Assuntos
Praguicidas , Ratos , Animais , Masculino , Praguicidas/toxicidade , Ratos Wistar , Dano ao DNA , Fígado , Modelos AnimaisRESUMO
The water deficit in particular, reduces the productivity of vegetable crops. To minimize these harmful effects on agriculture, several agronomic and physiological practices are being studied, such as the use of bacteria and water stress attenuators, such as brassinosteroids. Considering the socioeconomic relevance of corn culture and its sensitivity when exposed to water deficit, the objective of the present study was to evaluate the action of brassinosteroids and azospirillum on nitrogen metabolism in corn plants subjected to water stress conditions. The experiment was carried out in a greenhouse, in a period of 47 days, with corn plants, using the hybrid K9606 VIP3. The design was completely randomized, in a 2x2x3 factorial scheme, with six replications. The first factor corresponds to two water regimes (presence and absence of water deficit). The second corresponds to inoculation via seed of Azospirillum brasiliense and absence of inoculation. And the third corresponds to the application of three concentrations of brassinosteroids (0, 0.3 and 0.6 µM). Were determined Nitrate; nitrate reductase; free ammonium; total soluble aminoacids; soluble proteins; proline; glycine betaine and glutamine synthetase. The lack of water in plants provided a reduction in the protein and nitrate reductase contents, in leaves and roots. For ammonium, plants with water deficit inoculated at a concentration of 0.3 µM, obtained an increase of 7.16 (70.26%) and 13.89 (77.04%) mmol NH4 + .Kg-1. DM (Dry mass) on the leaf and root respectively. The water deficit in the soil provided significant increases in the concentrations of glycine betaine, nitrate, proline and aminoacids, both in the leaves and in the roots of the corn plants. On the other hand, the contents of glutamine synthetase had a reduction in both leaves and roots.
Assuntos
Compostos de Amônio , Azospirillum brasilense , Zea mays , Brassinosteroides/metabolismo , Nitratos , Raízes de Plantas/metabolismo , Secas , Desidratação/metabolismo , Betaína/metabolismo , Glutamato-Amônia Ligase , Aminoácidos/metabolismo , Prolina/metabolismo , Nitrato Redutases/metabolismo , Nitrogênio/metabolismoRESUMO
The article "Study of liver toxicity and DNA damage due to exposure to the pesticide Mancozeb in an experimental animal model - A pilot model", by N.D. Suarez Uribe, M.F. Pezzini, J. Dall'Agnol, N. Marroni, S. Benitez, D. Benedetti, J. Da Silva, C.T. Cerski, E. Dallegrave, S. Macedo, S.C.W.S.E.F. de Oliveira, D. Joveleviths, published in Eur Rev Med Pharmacol Sci 2023; 27 (13): 6374-6383-DOI: 10.26355/eurrev_202307_32997-PMID: 37458654 has been retracted by the Editor in Chief for the following reasons: After publication, concerns were raised by an unidentified reader who underlined some similarities between this publication and a previous publication published in the Journal of Clinical and Experimental Gastroenterology. After being informed, the authors claimed the previous journal published the article without consent, and, therefore, the authors promptly withdrew the previous publication. The retraction published by the other journal does not contain any information regarding the reason for withdrawal. As a matter of fact, the journal does not have any evidence about the authors' claim and still considers this research a duplicate publication. For the above-mentioned reasons, the Editor in Chief decided to withdraw the manuscript. This manuscript has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/32997.
RESUMO
This article presents an extensive study to select the theoretical probability distribution that represents the concentrations of major constituents from 35 wastewater treatment plants located in Brazil, as well as to verify the influence of the adopted treatment technology on the data distributions. Due to the lack of a solid basis for selecting the distributions that best fit the data, various distributions had to be tried and the choice was based on statistical tests and visual techniques, comprising a total of 3,444 tests undertaken. While comparing suitability of five distributions (normal, lognormal, gamma, exponential and rectangular) for analysis of data (influent and effluent biochemical oxygen demand, chemical oxygen demand, suspended solids, nitrogen, phosphorus, thermotolerant coliforms and flow), it was observed that the lognormal distribution was the most suitable, which is in agreement with the findings from other research studies. This conclusion requires a different position from the one currently adopted when analysing plant performance, in which symmetry of the data is generally implied, which has been shown not to be the case with the large array of data sets investigated.
Assuntos
Esgotos/química , Esgotos/microbiologia , Eliminação de Resíduos Líquidos/métodos , Purificação da Água/métodos , BrasilRESUMO
BACKGROUND: Here we describe a new class of cryptides (peptides encrypted within a larger protein) with antimicrobial properties, named schistocins, derived from SmKI-1, a key protein in Shistosoma mansoni survival. This is a multi-functional protein with biotechnological potential usage as a therapeutic molecule in inflammatory diseases and to control schistosomiasis. METHODS: We used our algorithm enCrypted, to perform an in silico proteolysis of SmKI-1 and a screening for potential antimicrobial activity. The selected peptides were chemically synthesized, tested in vitro and evaluated by both structural (CD, NMR) and biophysical (ITC) studies to access their structure-function relationship. RESULTS: EnCrypted was capable of predicting AMPs in SmKI-1. Our biophysical analyses described a membrane-induced conformational change from random coil-to-α-helix and a peptide-membrane equilibrium for all schistocins. Our structural data allowed us to suggest a well-known mode of peptide-membrane interaction in which electrostatic attraction between the cationic peptides and anionic membranes results in the bilayer disordering. Moreover, the NMR H/D exchange data with the higher entropic contribution observed for the peptide-membrane interaction showed that schistocins have different orientations upon the membrane. CONCLUSIONS: This work demonstrate the robustness for using the physicochemical features of predicted peptides in the identification of new bioactive cryptides. Besides, it demonstrates the relevance of combining these analyses with biophysical methods to understand the peptide-membrane affinity and improve further algorithms. GENERAL SIGNIFICANCE: Bioprospecting cryptides can be conducted through data mining of protein databases demonstrating the success of our strategy. The peptides-based agents derived from SmKI-1 might have high impact for system-biology and biotechnology.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Schistosoma mansoni/química , Animais , Antibacterianos/síntese química , Antibacterianos/química , Antifúngicos/síntese química , Antifúngicos/química , Candida/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Proteínas Citotóxicas Formadoras de Poros/síntese química , Proteínas Citotóxicas Formadoras de Poros/química , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacosRESUMO
Arterial hypertension is a risk factor for various cardiovascular and renal diseases, representing a major public health challenge. Although a wide range of treatment options are available for blood pressure control, many hypertensive individuals remain with uncontrolled hypertension. Thus, the search for new substances with antihypertensive potential becomes necessary. Coumarins, a group of polyphenolic compounds derived from plants, have attracted intense interest due to their diverse pharmacological properties, like potent antihypertensive activities. Braylin (6-methoxyseselin) is a coumarin identified in the Zanthoxylum tingoassuiba species, described as a phosphodiesterase-4 (PDE4) inhibitor. Although different coumarin compounds have been described as potent antihypertensive agents, the activity of braylin on the cardiovascular system has yet to be investigated. To investigate the vasorelaxation properties of braylin and its possible mechanisms of action, we performed in vitro studies using superior mesenteric arteries and the iliac arteries isolated from rats. In this study, we demonstrated, for the first time, that braylin induces potent vasorelaxation, involving distinct mechanisms from two different arteries, isolated from rats. A possible inhibition of phosphodiesterase, altering the cyclic adenosine monophosphate (cAMP)/cAMP-dependent protein kinase (PKA) pathway, may be correlated with the biological action of braylin in the mesenteric vessel, while in the iliac artery, the biological action of braylin may be correlated with increase of cyclic guanosine monophosphate (cGMP), followed by BKCa, Kir, and Kv channel activation. Together, these results provide evidence that braylin can represent a potential therapeutic use in preventing and treating cardiovascular diseases.
Assuntos
Cumarínicos/farmacologia , Artéria Ilíaca/efeitos dos fármacos , Artérias Mesentéricas/efeitos dos fármacos , Vasodilatadores/farmacologia , Animais , Artéria Ilíaca/fisiologia , Masculino , Artérias Mesentéricas/fisiologia , Canais de Potássio/fisiologia , Ratos Wistar , Vasodilatação/efeitos dos fármacosRESUMO
Schistosoma mansoni infection has been associated with protection against allergies. The mechanisms underlying this association may involve regulatory cells and cytokines. We evaluated the immune response induced by the S. mansoni antigens Sm22.6, PIII and Sm29 in a murine model of ovalbumin (OVA)-induced airway inflammation. BALB/c mice were sensitized with subcutaneously injected OVA-alum and challenged with aerolized OVA. Mice were given three doses of the different S. mansoni antigens. Lung histopathology, cellularity of bronchoalveolar lavage (BAL) and eosinophil peroxidase activity in lung were evaluated. Immunoglobulin (Ig)E levels in serum and cytokines in BAL were also measured. Additionally, we evaluated the frequency of CD4+forkhead box P3 (FoxP3)+ T cells in cultures stimulated with OVA and the expression of interleukin (IL)-10 by these cells. The number of total cells and eosinophils in BAL and the levels of OVA-specific IgE were reduced in the immunized mice. Also, the levels of IL-4 and IL-5 in the BAL of mice immunized with PIII and Sm22.6 were decreased, while the levels of IL-10 were higher in mice immunized with Sm22.6 compared to the non-immunized mice. The frequency of CD4+FoxP3+ T cells was higher in the groups of mice who received Sm22.6, Sm29 and PIII, being the expression of IL-10 by these cells only higher in mice immunized with Sm22.6. We concluded that the S. mansoni antigens used in this study are able to down-modulate allergic inflammatory mediators in a murine model of airway inflammation and that the CD4+FoxP3+ T cells, even in the absence of IL-10 expression, might play an important role in this process.
Assuntos
Alveolite Alérgica Extrínseca/imunologia , Antígenos de Helmintos/imunologia , Schistosoma mansoni/imunologia , Alveolite Alérgica Extrínseca/induzido quimicamente , Alveolite Alérgica Extrínseca/prevenção & controle , Animais , Asma , Líquido da Lavagem Broncoalveolar/química , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/imunologia , Citocinas/análise , Modelos Animais de Doenças , Feminino , Fatores de Transcrição Forkhead/análise , Imunização , Interleucinas/análise , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Ovalbumina/toxicidade , Eosinofilia Pulmonar/induzido quimicamente , Eosinofilia Pulmonar/imunologia , Eosinofilia Pulmonar/prevenção & controle , Subpopulações de Linfócitos T/química , Subpopulações de Linfócitos T/imunologiaRESUMO
BACKGROUND: Around 300 million people world-wide suffer from asthma, and the prevalence of allergic diseases has increased. Much effort has been used in the study of mechanisms involved in the immune response observed in asthma to intervene for the treatment of this condition. During inflammation in asthma, Th2 cytokines and eosinophils are essential components of the host immune system. Furthermore, for therapeutic interventions against this disease, IL-10 is an important cytokine because it has a central role in the regulation of inflammatory cascades. OBJECTIVE: To evaluate the immunomodulatory effect of Lactococcus lactis strains expressing recombinant IL-10 in a mouse model of ovalbumin (OVA)-induced acute airway inflammation. METHODS: L. lactis expressing recombinant IL-10 in a cytoplasmic (LL-CYT) or secreted form (LL-SEC) and wild-type (LL-WT) were used. IL-10 production by the recombinant strains was evaluated by ELISA. After an intranasal administration of L. lactis producing recombinant IL-10 and the induction of acute allergic airway inflammation in mice, blood samples were collected to detect IgE anti-OVA, and bronchoalveolar lavage (BAL) was harvested for eosinophil count. Additionally, the lungs were collected for the detection of the eosinophil peroxidase (EPO) activity, measurement of cytokines and chemokines and evaluation of pathology. RESULTS: Mice that received LL-CYT and LL-SEC strains showed a significant decrease in eosinophils numbers, EPO activity, anti-OVA IgE and IgG1 levels, IL-4 and CCL3 production and pulmonary inflammation and mucus hypersecretion, compared with the asthmatic group. Only the LL-CYT/OVA group showed reduced levels of IL-5, CCL2, CCL5 and CCL11. CONCLUSION: Treatment with L. lactis producing recombinant IL-10 used in this study (LL-CYT and LL-SEC) modulated experimental airway inflammation in the mouse model independently of Treg cells. Additionally, the LL-CYT strain was more efficient in the suppression of lung inflammation.
Assuntos
Terapia Genética/métodos , Hipersensibilidade/imunologia , Interleucina-10/biossíntese , Lactococcus lactis/genética , Pneumonia/imunologia , Administração Intranasal , Animais , Asma/imunologia , Asma/patologia , Separação Celular , Citocinas/análise , Citocinas/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Vetores Genéticos , Hipersensibilidade/patologia , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoterapia/métodos , Interleucina-10/genética , Interleucina-10/imunologia , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Pneumonia/patologia , Proteínas Recombinantes/imunologia , Células Th2/imunologiaRESUMO
Schistosomiasis is a parasitic disease with more than 200 million people infected worldwide. The formation of granulomas around eggs trapped in the liver is the main cause of disease morbidity. Therefore, the aim of this investigation was to characterize the immunopathological response induced by the recombinant (r) IPSE/alpha-1 egg protein in mice. Herein, we have shown that splenocytes from mice immunized with rIPSE/alpha-1 produced IFN-gamma, TNF-alpha, IL-4, IL-5 and IL-13 characterizing a mixed Th1/Th2 type of immune response. Pathological analysis of the liver revealed that there was no alteration in the number of eggs and granulomas; however, we observed an increase in granuloma area in immunized mice. Furthermore, eosinophil peroxidase assay showed that there was no alteration in the eosinophil infiltration in the liver; however, n-acetyl-beta-glucosaminidase measurement revealed an increase in macrophage activity. Despite the alteration in the profile of liver inflammatory cells in rIPSE immunized mice, the production of chemokines such as CCL2, CCL3, CCL5 and CCL11 was unaltered compared with the control group. In conclusion, IPSE/alpha-1 immunization induces a mixed Th1/Th2 type of immune response and enlargement of hepatic granuloma caused by an increased macrophage activity, but does not alter Th2 cytokines following infection.
Assuntos
Proteínas do Ovo/imunologia , Proteínas de Helminto/imunologia , Schistosoma mansoni/imunologia , Schistosoma mansoni/patogenicidade , Esquistossomose mansoni/patologia , Esquistossomose mansoni/parasitologia , Células Th2/imunologia , Fatores de Virulência/imunologia , Acetilglucosaminidase/metabolismo , Animais , Citocinas/metabolismo , Proteínas do Ovo/genética , Eosinófilos/imunologia , Feminino , Granuloma/imunologia , Granuloma/parasitologia , Granuloma/patologia , Proteínas de Helminto/genética , Leucócitos Mononucleares/imunologia , Fígado/parasitologia , Fígado/patologia , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Peroxidase/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Esquistossomose mansoni/imunologia , Baço/imunologia , Fatores de Virulência/genéticaRESUMO
The surface of the schistosomula is an important target for host immune system attack because the tegument represents the interface between host and parasite and thus is a potential candidate for the development of new intervention strategies. In this study, we evaluated the ability of schistosomula tegument (Smteg) to induce protection in mice. Immunization of mice with Smteg together with Freund adjuvant induced a Th1 type of immune response associated with a significant reduction in worm burden (43-48%), eggs trapped in the liver (65%), eggs eliminated in the faeces (59-60%) and granuloma number (41%). Lastly, during an in vitro study, worms from mice immunized with Smteg showed damage in the adult worm tegument and impaired egg laying.
Assuntos
Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Animais , Citocinas/biossíntese , Modelos Animais de Doenças , Fezes/parasitologia , Feminino , Adjuvante de Freund/administração & dosagem , Fígado/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Contagem de Ovos de Parasitas , Schistosoma mansoni/ultraestrutura , Esquistossomose mansoni/imunologia , Esquistossomose mansoni/parasitologia , Esquistossomose mansoni/patologia , Células Th1/imunologia , Vacinas de Subunidades Antigênicas/administração & dosagem , Vacinas de Subunidades Antigênicas/imunologiaRESUMO
Proteins associated with the schistosome tegument are of great importance for the development of new intervention strategies since they may be exposed on the surface of the parasite. Herein, we have isolated a cDNA clone encoding for the Schistosoma mansoni SmIg and its recombinant protein was tested as a potential vaccine candidate. Initially, its amino acid sequence was analysed by bioinformatics and shown to possess an N-terminal signal peptide, a C-terminal transmembrane helix, 4 glycosylation sites, an immunoglobulin conserved domain and 73% similarity with a hypothetical S. japonicum protein of unknown function. SmIg was produced by E. coli as a recombinant protein (rSmIg) and its protective effectiveness was evaluated against S. mansoni infection with 100 cercariae in a murine model. Mice immunized with rSmIg induced an immune response characterized by dominant IgG1 isotype and significant levels of IFN-gamma, TNF-alpha, IL-10 and IL-4. Although immunogenic, the recombinant vaccine failed to induce worm burden reduction when compared to the infected control group. However, rSmIg-immunized mice had significant reductions of liver granuloma volume and fibrosis content by 31.8% and 49%, respectively. In conclusion, SmIg is a new tegument protein from S. mansoni that plays an important role in reducing pathology induced by parasite infection.
Assuntos
Antígenos de Helmintos/administração & dosagem , Proteínas de Helminto/administração & dosagem , Fígado/imunologia , Fígado/patologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Vacinas de DNA/administração & dosagem , Sequência de Aminoácidos , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Feminino , Proteínas de Helminto/química , Proteínas de Helminto/genética , Proteínas de Helminto/imunologia , Imunização , Imunoglobulina G/sangue , Camundongos , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Esquistossomose mansoni/imunologia , Vacinas de DNA/genética , Vacinas de DNA/imunologiaRESUMO
In Portugal landslides caused 237 fatalities and >1600 displaced people in the period 1865-2015. Spatial distribution and temporal patterns of slope instability can be related with a complex set of natural and human factors responsible for generating damages. It is essential to develop new methodologies to synthetize risk dimensions to contribute to the landslide risk management at the municipal level. This work proposed a municipal landslide risk index (LRI) considering three risk dimensions: hazard, exposure and physical vulnerability of buildings. The hazard dimension includes the landslide susceptibility performed at the national scale, the probability of weather types associated with landslides and an extreme precipitation susceptibility index. The exposure dimension considered the population density and the number of buildings, whereas the average physical vulnerability of the buildings was computed using four statistical variables from the official census: (i) construction technique and construction materials; (ii) reinforced structure; (iii) number of floors; and (iv) conservation status. Each variable includes different classes that were empirically weighted. After evaluating the three risk dimensions and the LRI, a cluster analysis was performed in order to identify the most important landslide risk drivers in each municipality. Exposure is the main driving force of LRI in the metropolitan areas of Lisbon and Porto, while the hazard is more relevant in the NW municipalities and the physical vulnerability is the major driving force in the south of the country. This methodological approach contributes to a comprehensive and synthetized knowledge about the landslide risk driving forces within the 278 Portuguese municipalities. In addition, it contributes to the diversification and context-oriented strategies of landslide risk management that still lacks in most of the national-level risk governance processes. Finally, this methodology can be generalized to other geographical contexts, improving the risk management, land use planning and the disaster risk reduction.
RESUMO
Sequence databases on Schistosoma mansoni have revealed micro-exon gene (MEGs) families. Many of these genes are highly expressed in parasite life cycle stages associated with the mammalian host infection and appear to be involved in immune evasion by schistosomes. So, we believe that MEG-coding proteins would make potential candidates for vaccine development or diagnosis for schistosomiasis. Here, we study MEG-3.2 and MEG-3.4, members of the MEG-3 family. Recombinant (r) proteins were produced and formulated with Freund's adjuvant for vaccination of mice. Immunization with recombinant MEG-3.2 or MEG-3.4 formulation generated high levels of IgG1 antibodies. Additionally, vaccination also induced a mixed Th1/Th2/Th17-type of response, since IFN-γ, IL-5 and IL-17 cytokines were detected in the supernatant of spleen cell cultures; however it failed to induce reduction in parasitic worm burden. Finally, the recombinant proteins were evaluated in a serological assay using human samples. Schistosome-infected individuals showed higher levels of both IgG and IgM against rMEG-3.2 compared to non-infected individuals, while only IgM anti-rMEG-3.4 antibodies were elevated in infected patients. Therefore, between both studied molecules, MEG-3.2 protein is the antigen that shows potential to compose a serological diagnosis test for schistosomiasis.
Assuntos
Antígenos de Helmintos/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/prevenção & controle , Animais , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Esquistossomose mansoni/sangue , Esquistossomose mansoni/diagnóstico , VacinaçãoRESUMO
Debris flows are one of the most hazardous types of landslides in mountain regions. In the upper part of the Zêzere valley (Serra da Estrela, Portugal) several debris flows events occurred in the last 200 years, some of them causing loss of lives and material damages. In this work, a methodology for pedestrian evacuation modelling, in a debris flow hazard scenario, was implemented. A dynamic run-out model, developed in previous studies, was used to evaluate the debris flows velocities, thickness of the deposits and extent of the mobilized material. The buildings potentially affected by the impact of debris flows were identified and the potentially exposed population was estimated by applying a dasymetric distribution. The results lead to the conclusion that, in the study area, the elderly are those who are most exposed to debris flows. Furthermore, the time lapse between the debris flows initiation and the arrival at the buildings at risk was estimated, allowing to account for the overall number of buildings where the evacuation time takes longer than the debris flows arrival. Additionally, the safe areas within the study area were identified, and several safe public buildings with the capacity to gather a large number of persons were selected. Considering that the study area is located in a mountain region, characterized by steep slopes, the evacuation modelling was performed based on an anisotropic approach, in order to consider the influence of slope direction on travel costs. At the end, three pedestrian evacuation travel time scenarios, based on different walking speeds to accommodate residents with different ages in safer places, were compared and the results mapped. The implemented methodology is not local dependent, which allows its reproduction elsewhere.
RESUMO
This work reports the structural and enzymatic characterization of a new sPLA2 from the white venom of Crotalus durissus ruruima, nominated PLA2A. The homogeneity of the PLA2A fraction and its molecular mass were initially evaluated by SDS-PAGE and confirmed by MALDI-TOF spectrometry, indicating a molecular mass of 14,299.34Da. Structural investigation, through circular dichroism spectroscopy, revealed that PLA2A has a high content of alpha helix and beta-turn structures, 45.7% and 35.6% respectively. Its amino acid sequence, determined by Edman degradation and "de novo amino acid sequencing", exhibited high identity to PLA2 Cdt F15 from Crotalus durissus terrificus. The enzymatic investigation, conducted using the synthetic substrate 4-nitro-3-(octanoyloxy)-benzoic acid, determined its V(max) (7.56nmoles/min) and K(M) (2.76mM). Moreover, PLA2A showed an allosteric behavior and its enzymatic activity was dependent on Ca(2+). Intrinsic fluorescence measurements suggested that Ca(2+) induced a significant increase of PLA2A fluorescence, whereas its replacement for Mg(2+), Mn(2+), Sn(2+) and Cd(2+) apparently induced no structural modifications. The optimal pH and temperature for the enzymatic activity of PLA2A were 8.4 and 40 degrees C, respectively, and the minimal concentration of p-BPB and crotapotin that significantly inhibited such activity was 0.75mM and 0.4muM, respectively. In addition, PLA2A showed a significant antibacterial effect that was not strictly dependent on the enzymatic activity of such sPLA2.