Epidemiological characterization of resistance and PCR typing of Shigella flexneri and Shigella sonnei strains isolated from bacillary dysentery cases in Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-motile, and non-sporulated bacilli of the Enterobacteriaceae family responsible for "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S. flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3% of cases, chloramphenicol in 70.0%, streptomycin in 86.7%, sulfamethoxazole in 80.0%, and tetracycline in 80.0%, while a smaller number of strains were resistant to cephalothin (3.3%) and sulfazotrim (10.0%). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0%) and tetracycline (96.7%) and, to a lesser extent, to ampicillin (6.7%) and streptomycin (26.7%). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved

Epidemiological Characterization Of Resistance And Pcr Typing Of Shigella Flexneri And Shigella Sonnei Strains Isolated From Bacillary Dysentery Cases In Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-montile, and non-sporulated bacilli of the Enterobacteriaceae family responsible or "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S.flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3% of cases, chloramphenicol in 70.0%, streptomycin in 86.7%, sulfamethoxazole in 80.0%, and tetracycline in 80.0%, while a smaller number of strains were resistant to cephalothin (3.3%) and sulfazotrim (10.0%). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0%) and tetracycline (96.7%) and, to a lesser extent, to ampicillin (6.7%) and streptomycin (26.7%). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved.402249258Murray, P.R., Rosenthal, K.S., Kobayashi, G.S., Pfaller, M.A., (1998) Medical Microbiology, , 3rd edn. 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Use Of Mesoporous Silica Sba-15 And Sba-16 In Association Of Outer Membrane Vesicles - Omv From Neisseria Meningitidis

Outer membrane vesicles or OMV are nanoparticles released in culture medium during meningococcal growth resulting from evaginations of the outer cellular membrane and have been indicated as potential target for vaccine production. This study aimed to analyze the use of Neisseria meningitidis B2443, as vaccine using a semi-solid fermentation process based in ultrafiltration for the isolation these OMV and also verify the effect of the mesouporous silica (SBA-15 and SBA-16). The OMV preparation follow the method without the ultracentrifugation whose was subtituted by ultrafiltration method using a nitrocelulosis filtre showing a pore of 0.025 μm. For the detection of antibodies production were used the immunological method of ELISA, and serum bactericidal effect using sera from immunized mices with OMV and adjuvant inorganic nanoparticles. Also, the use of citotocity test were performed based in the neutral red uptake for safety of the associated vaccin use in NIH-3T3 cell line. It was compared to OMV production of strains of N. meningitidis strains B2443 and C2135. The results showed that different strains of N. meningitidis have OMVs kinetics of production of different time and quantity. The use of SBA-15 and SBA-16 as adujvant at 250 μg for each mice was enough to induce an increase of vaccinal (for other serogroups) capacity same using a only OMV extracted from strains B2443. The study showed that the methodology used for the production of OMV is advantageous from the point of view of quantity and cost and use of this biologic nanoparticle. Both mesoporous silica SBa15 and SBa16 used in this work were capable to increase de recognition of antibody against different strains fo N. meningitidis showed using the OMV extracted from an only vaccinal strain. © 2013 Alves DA, et al.46Frasch, C.E., Production and control of Neisseria meningitidis vaccines (1990) Adv Biotechnol Processes, 13, pp. 123-145Frasch, C.E., Sacchi, C.T., Brandiolone, M.C., Vieiera, V.S., Leite, L.C., Development of a second generation group B meningococcal vaccine (1991) NIPH Ann, 14, pp. 225-230Comanducci, M., Bambini, S., Brunelli, B., Adu-Bobie, J., Aricò, B., NadA, a novel vaccine candidate of Neisseria meningitidis (2002) J Exp Med, 195, pp. 1445-1454Borrow, R., Balmer, P., Miller, E., Meningococcal surrogates of protection--serum bactericidal antibody activity (2005) Vaccine, 23, pp. 2222-2227Milagres, L.G., Ramos, S.R., Sacchi, C.T., Melles, C.E., Vieira, V.S., Immune response of Brazilian children to a Neisseria meningitidis serogroup B outer membrane protein vaccine: Comparison with efficacy (1994) Infect Immun, 62, pp. 4419-4424Milagres, L.G., Gorla, M.C., Sacchi, C.T., Rodrigues, M.M., Specificity of bactericidal antibody response to serogroup B meningococcal strains in Brazilian children after immunization with an outer membrane vaccine (1998) Infect Immun, 66, pp. 4755-4761Frederiksen, W., Ecology and significance of Pasteurellaceae in man--an update (1993) Zentralbl Bakteriol, 279, pp. 27-34Zollinger, W.D., Moran, E.E., Devi, S.J., Frasch, C.E., Bactericidal antibody responses of juvenile rhesus monkeys immunized with group B Neisseria meningitidis capsular polysaccharide-protein conjugate vaccines (1997) Infect Immun, 65, pp. 1053-1060Morley, S.L., Pollard, A.J., Vaccine prevention of meningococcal disease, coming soon? (2001) Vaccine, 20, pp. 666-687Jódar, L., Feavers, I.M., Salisbury, D., Granoff, D.M., Development of vaccines against meningococcal disease (2002) Lancet, 359, pp. 1499-1508Pollard, A.J., Moxon, E.R., The meningococcus tamed? 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Pathogenic And Opportunistic Respiratory Bacteria-induced Apoptosis

Several pathogenic or opportunistic bacteria have the ability to either induce or inhibit host cell apoptosis. The capacity to modulate cell pathways that result in the induction or delay of host cell apoptosis is considered to be an important bacterial virulence mechanism. These processes could be mediated by different host cell signaling pathways that are subverted by the bacteria. Pathogens are able to activate apoptotic proteins, such as caspases, or inactivate anti-apoptotic proteins, such as NFkB and the MAPKKs, or even up-regulate the endogenous receptor/ ligand system that induces apoptosis, generally when the bacteria are bound to the host cell surface. The bacteria-induced apoptotic or anti-apoptotic processes are often related with the fact that the bacteria acquire the ability to reach the host tissues. However, apoptosis is also considered to be a host defense mechanism against infectious agents. 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Membrane Protein As Novel Targets For Vaccine Production In Haemophilus Influenzae And Neisseria Meningitidis

Haemophilus influenzae and Neisseria meningitis are gram negative, commensal bacteria naturally present in the nasopharynx. They are also naturally competent and suffer genetic mutations. H. influenzae causes diseases such as otitis media and pneumonia. While N. meningitis causes pneumonia, meningitis and sepsis. With the introduction of a vaccination program, a decrease in cases and deaths caused by these pathogens were observed over the following years. Especially, in countries where these vaccines were included in the vaccination schedule and in endemic regions, such as the meningitis belt in Africa. However, there are serotypes, biotypes and strains that these vaccines do not cover. Thus, these strains, biotypes and serotypes are emerging as pathogenic ones. Concerning the health authorities because the diagnosis of these diseases is most of the times unreliable and treatment needs to be immediate, due to the rapid evolution of the disease. For these emerging bacteria novel immunogenic targets are being researched as a way of trying to find and design new vaccines. These vaccines can be aimed in membrane proteins. Using these proteins as immunogenic targets with the help of adjuvants, to boost the immune system. The burden of a death or sequel to children and even adults that undergo meningitis infection and treatment is high. Therefore prevention is the best alternative. The aim of this review is to present this novel targets and their pros and cons. 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immunization programmes, , World Health Organization, Management guidelines, including information for health workers and parents. 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A cost-effectiveness model of strategy assessment (1985) JAMA, 253, pp. 521-529Baggett, H.C., Hennessy, T.W., Bulkow, L., Romero-Steiner, S., Hurlburt, D., Immunologic response to Haemophilus influenzae type b (Hib) conjugate vaccine and risk factors for carriage among Hib carriers and noncarriers in Southwestern Alaska (2006) Clin Vaccine Immunol, 13, pp. 620-626(2009) Haemophilus b Conjugate Vaccine, , Confidential/Proprietary Information, (Tetanus Toxoid Conjugate) ActHIB®Liquid PedvaxHIB®, , [Haemophilus b Conjugate Vaccine (Meningococcal Protein Conjugate)]Barenkamp, S.J., Munson Jr., R.S., Granoff, D.M., Subtyping isolates of Haemophilus influenzae type b by outer-membrane protein profiles (1981) J Infect Dis, 143, pp. 668-676Foxwell, A.R., Kyd, J.M., Cripps, A.W., Nontypeable Haemophilus influenzae: Pathogenesis and prevention (1998) Microbiol Mol Biol Rev, 62, pp. 294-308Murphy, T.F., Bartos, L.C., Purification and analysis with monoclonal antibodies of P2, the major outer 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A., Kaur, R., Michel, L.V., Casey, J.R., Pichichero, M., Haemophilus influenzae vaccine candidate outer membrane protein P6 is not conserved in all strains (2011) Hum Vaccin, 7, pp. 102-105Magagnoli, C., Bardotti, A., De Conciliis, G., Galasso, R., Tomei, M., Structural organization of NadADelta(351-405), a recombinant MenB vaccine component, by its physico-chemical characterization at drug substance level (2009) Vaccine, 27, pp. 2156-2170Lucidarme, J., Newbold, L.S., Findlow, J., Gilchrist, S., Gray, S.J., Molecular targets in meningococci: Efficient routine characterization and optimal outbreak investigation in conjunction with routine surveillance of the meningococcal group B vaccine candidate, fHBP (2011) Clin Vaccine Immunol, 18, pp. 194-202Frasch, C.E., van Alphen, L., Holst, J., Poolman, J.T., Rosenqvist, E., Outer membrane protein vesicle vaccines for meningococcal disease (2001) Methods Mol Med, 66, pp. 81-107Giuliani, M.M., Adu-Bobie, J., Comanducci, M., Aricò, B., Savino, 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Graphene And Carbon Nanotube Nanocomposite For Gene Transfection

Graphene and carbon nanotube nanocomposite (GCN) was synthesised and applied in gene transfection of pIRES plasmid conjugated with green fluorescent protein (GFP) in NIH-3T3 and NG97 cell lines. The tips of the multi-walled carbon nanotubes (MWCNTs) were exfoliated by oxygen plasma etching, which is also known to attach oxygen content groups on the MWCNT surfaces, changing their hydrophobicity. The nanocomposite was characterised by high resolution scanning electron microscopy; energy-dispersive X-ray, Fourier transform infrared and Raman spectroscopies, as well as zeta potential and particle size analyses using dynamic light scattering. BET adsorption isotherms showed the GCN to have an effective surface area of 38.5 m2/g. The GCN and pIRES plasmid conjugated with the GFP gene, forming π-stacking when dispersed in water by magnetic stirring, resulting in a helical wrap. The measured zeta potential confirmed that the plasmid was connected to the nanocomposite. The NIH-3T3 and NG97 cell lines could phagocytize this wrap. The gene transfection was characterised by fluorescent protein produced in the cells and pictured by fluorescent microscopy. Before application, we studied GCN cell viability in NIH-3T3 and NG97 line cells using both MTT and Neutral Red uptake assays. Our results suggest that GCN has moderate stability behaviour as colloid solution and has great potential as a gene carrier agent in non-viral based therapy, with low cytotoxicity and good transfection efficiency. © 2014 Elsevier B.V.391288298Vardharajula, S., Ali, S.Z., Tiwari, P.M., Eroglu, E., Vig, K., Dennis, V.A., Functionalized carbon nanotubes: Biomedical applications (2012) Int. J. 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Carbon nanotubes hoax scientists in viability assays (2006) Nano Lett., 6, pp. 1261-1268Casey, A., Davoren, M., Herzog, E., Lyng, F.M., Byrne, H.J., Chambers, G., Probing the interaction of single walled carbon nanotubes within cell culture medium as a precursor to toxicity testing (2007) Carbon, 45, pp. 34-40Casey, A., Herzog, E., Davoren, M., Lyng, F.M., Byrne, H.J., Chambers, G., Spectroscopic analysis confirms the interactions between single walled carbon nanotubes and various dyes commonly used to assess cytotoxicity (2007) Carbon, 45, pp. 1425-1432Hurt, R.H., Monthioux, M., Kane, A., Toxicology of carbon nanomaterials: Status, trends, and perspectives on the special issue (2006) Carbon, 44, pp. 1028-1033Zanin, H., Peterlevitz, A.C., Ceragioli, H.J., Rodrigues, A.A., Belangero, W.D., Baranauskas, V., Magnetic and cytotoxic properties of hot-filament chemical vapour deposited diamond (2012) Mater. Sci. Eng. C-Mate. Biol. 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Advances In Nanobiomaterials For Oncology Nanomedicine

The word "cancer" is generically used to identify a wide range of diseases that are malignant tumors. Malignant tumors are of very different types, with different existing causes, evolutions, and thus with different treatments for each type. However, there is one feature that is common to all tumors: the division and uncontrolled growth of cells. The causes of cancer are not fully known, however, the modification of DNA appears to be responsible for the alteration of normal cell growth. Some chemicals can also induce the formation of cancerous tumors. In recent times, there has been considerable progress in the development of therapeutic drugs that act specifically on detectable molecular abnormalities in certain tumors, minimizing damage to normal cells. The prognosis for patients with cancer is influenced by the type of cancer; however, the grading and the presence of specific molecular markers can also bring benefits to establish the prognosis and determining individual treatments. This chapter focuses on the recent advances in nanobiomaterials (i.e., nanoparticles produced by biomaterials), used for the development of innovative approaches for oncology. Nanobiomaterials may be applied for cancer treatment either by passive or active pathways. In the passive pathway, nanoparticles circulate in the bloodstream and are accumulated into the tumor through enhanced permeability and retention effect; in the active pathway, nanoparticles reach the tumor through targeted drug therapy. Several types of biomaterials (e.g., lipids, phospholipids, polymers) may be used for the production of these nanoparticles, with specific size and surface electrical charge, and may even be surfaced with specific targeting ligands for site-specific targeting. © 2016 Elsevier Inc. All rights reserved.9111

Chemical Modifications Of Phtx-i Myotoxin From Porthidium Hyoprora Snake Venom: Effects On Structural, Enzymatic, And Pharmacological Properties

We recently described the isolation of a basic PLA2 (PhTX-I) from Porthidium hyoprora snake venom. This toxin exhibits high catalytic activity, induces in vivo myotoxicity, moderates footpad edema, and causes in vitro neuromuscular blockade. Here, we describe the chemical modifications of specific amino acid residues (His, Tyr, Lys, and Trp), performed in PhTX-I, to study their effects on the structural, enzymatic, and pharmacological properties of this myotoxin. After chemical treatment, a single His, 4 Tyr, 7 Lys, and one Trp residues were modified. The secondary structure of the protein remained unchanged as measured by circular dichroism; however other results indicated the critical role played by Lys and Tyr residues in myotoxic, neurotoxic activities and mainly in the cytotoxicity displayed by PhTX-I. His residue and therefore catalytic activity of PhTX-I are relevant for edematogenic, neurotoxic, and myotoxic effects, but not for its cytotoxic activity. This dissociation observed between enzymatic activity and some pharmacological effects suggests that other molecular regions distinct from the catalytic site may also play a role in the toxic activities exerted by this myotoxin. 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