RESUMEN
Malaria remains a global health challenge, necessitating the development of effective vaccines. The RTS,S vaccination prevents Plasmodium falciparum (Pf) malaria but is ineffective against Plasmodium vivax (Pv) disease. Herein, we evaluated the murine immunogenicity of a recombinant PvCSP incorporating prevalent polymorphisms, adjuvanted with Alhydrogel or Poly I:C. Both formulations induced prolonged IgG responses, with IgG1 dominance by the Alhydrogel group and high titers of all IgG isotypes by the Poly I:C counterpart. Poly I:C-adjuvanted vaccination increased splenic plasma cells, terminally-differentiated memory cells (MBCs), and precursors relative to the Alhydrogel-combined immunization. Splenic B-cells from Poly I:C-vaccinated mice revealed an antibody-secreting cell- and MBC-differentiating gene expression profile. Biological processes such as antibody folding and secretion were highlighted by the Poly I:C-adjuvanted vaccination. These findings underscore the potential of Poly I:C to strengthen immune responses against Pv malaria.
Asunto(s)
Adyuvantes de Vacunas , Hidróxido de Aluminio , Inmunogenicidad Vacunal , Vacunas contra la Malaria , Malaria Vivax , Plasmodium vivax , Poli I-C , Proteínas Protozoarias , Poli I-C/administración & dosificación , Plasmodium vivax/inmunología , Inmunidad Humoral , Inmunidad Celular , Proteínas Protozoarias/inmunología , Vacunas contra la Malaria/química , Vacunas contra la Malaria/inmunología , Hidróxido de Aluminio/administración & dosificación , Inmunoglobulina G/sangre , Masculino , Animales , Células Plasmáticas/inmunología , Femenino , Ratones Endogámicos C57BL , Proteínas Recombinantes/inmunología , Vacunación , Adyuvantes de Vacunas/administración & dosificación , Malaria Vivax/prevención & controlRESUMEN
Breast cancer is the most incidental and deadly neoplasm worldwide; in Mexico, very few epidemiologic reports have analyzed the pathological features and its impact on their clinical outcome. Here, we studied the relation between pathological features and the clinical presentation at diagnosis and their impact on the overall and progression-free survival of patients with breast cancer. For this purpose, we collected 199 clinical records of female patients, aged at least 18 years old (y/o), with breast cancer diagnosis confirmed by biopsy. We excluded patients with incomplete or conflicting clinical records. Afterward, we performed an analysis of overall and progression-free survival and associated risks. Our results showed an average age at diagnosis of 52 y/o (24-85), the most common features were: upper outer quadrant tumor (32%), invasive ductal carcinoma (76.8%), moderately differentiated (44.3%), early clinical stages (40.8%), asymptomatic patients (47.8%), luminal A subtype (47.8%). Median overall survival was not reached, but median progression-free survival was 32.2 months (29.75-34.64, CI 95%) associated risk were: clinical stage (p < 0.0001) symptomatic presentation (p = 0.009) and histologic grade (p = 0.02). Therefore, we concluded that symptom presence at diagnosis impacts progression-free survival, and palpable symptoms are related to an increased risk for mortality.
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Neoplasias de la Mama , Carcinoma Ductal de Mama , Adulto , Femenino , Humanos , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , México/epidemiología , Estadificación de Neoplasias , Pronóstico , Estudios Retrospectivos , Adulto Joven , Persona de Mediana Edad , Anciano , Anciano de 80 o más AñosRESUMEN
OBJECTIVES: The purpose of this study was (i) to investigate whether nanocomposite poly(methyl-methacrylate)-zinc oxide nanowires (PMMA-ZnO-NWs) have C. albicans antibiofilm activity; (ii) to evaluate the interaction between components of the nanocomposites based on PMMA-ZnO-NWs by Raman spectroscopy; and (iii) to assess ultrastructural alterations. DESIGN: Sixty-eight rectangles (17 PMMA (control) and 51 PMMA-ZnO-NWs (250, 500, 1000 ppm ZnO nanowires) were fabricated. C. albicans ATCC 10231 and a C. albicans clinical strain were tested. Adherence, biofilm formation and ultrastructural alterations were assessed by transmission electron microscopy. Raman mapping images and spectra were analyzed using main component analysis. RESULTS: Nanocomposite PMMA-ZnO-NWs inhibited the formation of C. albicans biofilms 94% at 1000 ppm and 80% at 500 ppm against both C. albicans strains. PMMA-ZnO-NWs induced ultrastructural alterations, including cell wall damage and disorganization of the cytoplasmic membrane, resulting in cell lysis. Raman spectroscopy showed new vibrational modes (300-365-485-600 cm-1) for PMMA and ZnO-NW interactions. CONCLUSIONS: PMMA-ZnO-NWs inhibited C. albicans dose-dependent biofilm formation and led to changes in the structures and cell membrane. Raman spectroscopy showed chemical interactions between ZnO-NWs and PMMA, as suggested by the appearance of new bands at 301 and 485 cm-1.
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Two years ago, we held an exciting event entitled the São Paulo School of Advanced Sciences on Vaccines (SPSASV). Sixty-eight Ph.D. students, postdoctoral fellows and independent researchers from 37 different countries met at the Mendes Plaza Hotel located in the city of Santos, SP - Brazil to discuss the challenges and the new frontiers of vaccinology. The SPSASV provided a critical and comprehensive view of vaccine research from basics to the current state-of-the-art techniques performed worldwide. For 10 days, we discussed all the aspects of vaccine development in 36 lectures, 53 oral presentations and 2 poster sessions. At the end of the course, participants were further encouraged to present a model of a grant proposal related to vaccine development against individual pathogens. Among the targeted pathogens were viruses (Chikungunya, HIV, RSV, and Influenza), bacteria (Mycobacterium tuberculosis and Streptococcus pyogenes), parasites (Plasmodium falciparum or Plasmodium vivax), and the worm Strongyloides stercoralis. This report highlights some of the knowledge shared at the SPSASV.
RESUMEN
Plasmodium vivax is the most common species of human malaria parasite found outside Africa, with high endemicity in Asia, Central and South America, and Oceania. Although Plasmodium falciparum causes the majority of deaths, P. vivax can lead to severe malaria and result in significant morbidity and mortality. The development of a protective vaccine will be a major step toward malaria elimination. Recently, a formulation containing the three allelic variants of the P. vivax circumsporozoite protein (PvCSP-All epitopes) showed partial protection in mice after a challenge with the hybrid Plasmodium berghei (Pb) sporozoite, in which the PbCSP central repeats were replaced by the VK210 PvCSP repeats (Pb/Pv sporozoite). In the present study, the chimeric PvCSP allelic variants (VK210, VK247, and P. vivax-like) were fused with the mumps virus nucleocapsid protein in the absence (NLP-CSPR) or presence of the conserved C-terminal (CT) domain of PvCSP (NLP-CSPCT). To elicit stronger humoral and cellular responses, Pichia pastoris yeast was used to assemble them as nucleocapsid-like particles (NLPs). Mice were immunized with each recombinant protein adjuvanted with Poly (I:C) and presented a high frequency of antigen-specific antibody-secreting cells (ASCs) on days 5 and 30, respectively, in the spleen and bone marrow. Moreover, high IgG titers against all PvCSP variants were detected in the sera. Later, these immunized mice with NLP-CSPCT were challenged with Pb/Pv sporozoites. Sterile protection was observed in 30% of the challenged mice. Therefore, this vaccine formulation use has the potential to be a good candidate for the development of a universal vaccine against P. vivax malaria.
RESUMEN
Two years ago, we held an exciting event entitled the São Paulo School of Advanced Sciences on Vaccines (SPSASV). Sixty-eight Ph.D. students, postdoctoral fellows and independent researchers from 37 different countries met at the Mendes Plaza Hotel located in the city of Santos, SP - Brazil to discuss the challenges and the new frontiers of vaccinology. The SPSASV provided a critical and comprehensive view of vaccine research from basics to the current state-of-the-art techniques performed worldwide. For 10 days, we discussed all the aspects of vaccine development in 36 lectures, 53 oral presentations and 2 poster sessions. At the end of the course, participants were further encouraged to present a model of a grant proposal related to vaccine development against individual pathogens. Among the targeted pathogens were viruses (Chikungunya, HIV, RSV, and Influenza), bacteria (Mycobacterium tuberculosis and Streptococcus pyogenes), parasites (Plasmodium falciparum or Plasmodium vivax), and the worm Strongyloides stercoralis. This report highlights some of the knowledge shared at the SPSASV.(AU)
Asunto(s)
Escuelas para Profesionales de Salud , Vacunas/historia , Organización Mundial de la Salud , Antígenos , Esquemas de InmunizaciónRESUMEN
Anthropic activity in Antarctica has been increasing considerably in recent years, which could have an important impact on the local microbiota affecting multiple features, including the bacterial resistome. As such, our study focused on determining the antibiotic-resistance patterns and antibiotic-resistance genes of bacteria recovered from freshwater samples collected in areas of Antarctica under different degrees of human influence. Aerobic heterotrophic bacteria were subjected to antibiotic susceptibility testing and PCR. The isolates collected from regions of high human intervention were resistant to several antibiotic groups, and were mainly associated with the presence of genes encoding aminoglycosides-modifying enzymes (AMEs) and extended-spectrum ß-lactamases (ESBLs). Moreover, these isolates were resistant to synthetic and semi-synthetic drugs, in contrast with those recovered from zones with low human intervention, which resulted highly susceptible to antibiotics. On the other hand, we observed that zone A, under human influence, presented a higher richness and diversity of antibiotic-resistance genes (ARGs) in comparison with zones B and C, which have low human activity. Our results suggest that human activity has an impact on the local microbiota, in which strains recovered from zones under anthropic influence were considerably more resistant than those collected from remote regions.
Asunto(s)
Farmacorresistencia Microbiana , Agua Dulce/microbiología , Microbiología del Agua , Animales , Regiones Antárticas , Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Bacterias/genética , Farmacorresistencia Bacteriana , Ambiente , Genes Bacterianos , Geografía , Humanos , Pruebas de Sensibilidad Microbiana , Microbiota/efectos de los fármacos , Reacción en Cadena de la Polimerasa , beta-Lactamasas/metabolismoRESUMEN
Two years ago, we held an exciting event entitled the São Paulo School of Advanced Sciences on Vaccines (SPSASV). Sixty-eight Ph.D. students, postdoctoral fellows and independent researchers from 37 different countries met at the Mendes Plaza Hotel located in the city of Santos, SP - Brazil to discuss the challenges and the new frontiers of vaccinology. The SPSASV provided a critical and comprehensive view of vaccine research from basics to the current state-of-the-art techniques performed worldwide. For 10 days, we discussed all the aspects of vaccine development in 36 lectures, 53 oral presentations and 2 poster sessions. At the end of the course, participants were further encouraged to present a model of a grant proposal related to vaccine development against individual pathogens. Among the targeted pathogens were viruses (Chikungunya, HIV, RSV, and Influenza), bacteria (Mycobacterium tuberculosis and Streptococcus pyogenes), parasites (Plasmodium falciparum or Plasmodium vivax), and the worm Strongyloides stercoralis. This report highlights some of the knowledge shared at the SPSASV.(AU)
Asunto(s)
Instituciones Académicas , Vacunas , Técnicas Inmunológicas/métodos , Informe de Investigación , Vacunología , Concentración de Iones de HidrógenoRESUMEN
Salmon farming industry in Chile currently uses a significant quantity of antimicrobials to control bacterial pathologies. The main aims of this study were to investigate the presence of transferable sulfonamide- and trimethoprim-resistance genes, sul and dfr, and their association with integrons among bacteria associated to Chilean salmon farming. For this purpose, 91 Gram-negative strains resistant to sulfisoxazole and/or trimethoprim recovered from various sources of seven Chilean salmonid farms and mainly identified as belonging to the Pseudomonas genus (81.0%) were studied. Patterns of antimicrobial resistance of strains showed a high incidence of resistance to florfenicol (98.9%), erythromycin (95.6%), furazolidone (90.1%) and amoxicillin (98.0%), whereas strains exhibited minimum inhibitory concentrations (MIC90) values of sulfisoxazole and trimethoprim of >4,096 and >2,048 µg mL-1, respectively. Strains were studied for their carriage of these genes by polymerase chain reaction, using specific primers, and 28 strains (30.8%) were found to carry at least one type of sul gene, mainly associated to a class 1 integron (17 strains), and identified by 16S rRNA gene sequencing as mainly belonging to the Pseudomonas genus (21 strains). Of these, 22 strains carried the sul1 gene, 3 strains carried the sul2 gene, and 3 strains carried both the sul1 and sul2 genes. Among these, 19 strains also carried the class 1 integron-integrase gene intI1, whereas the dfrA1, dfrA12 and dfrA14 genes were detected, mostly not inserted in the class 1 integron. Otherwise, the sul3 and intI2 genes were not found. In addition, the capability to transfer by conjugation these resistance determinants was evaluated in 22 selected strains, and sul and dfr genes were successfully transferred by 10 assayed strains, mainly mediated by a 10 kb plasmid, with a frequency of transfer of 1.4 × 10-5 to 8.4 × 10-3 transconjugant per recipient cell, and exhibiting a co-transference of resistance to florfenicol and oxytetracycline, currently the most used in Chilean salmon industry, suggesting an antibacterial co-selection phenomenon. This is the first report of the characterization and transferability of integrons as well as sul and dfr genes among bacteria associated to Chilean salmon farms, evidencing a relevant role of this environment as a reservoir of these genes.
RESUMEN
Carbapenemase-producing Enterobacteriaceae have rapidly disseminated worldwide and can colonize patients in healthcare centers. As in Chile the first isolations of NDM-1 and OXA-370 carbapenemases were related with a patient arriving from Brazil, the genetic relatedness of Klebsiella pneumoniae strains producers of these enzymes and isolated in both countries was assessed. PFGE analyses revealed that the isolates were clonally related, illustrating how travel contributes to the spread of multidrug-resistant microorganisms. In addition, the occurrence of three different carbapenemases in three different K. pneumoniae strains isolated from a single patient is described.
Asunto(s)
Proteínas Bacterianas/genética , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/genética , beta-Lactamasas/genética , Antibacterianos , Brasil/epidemiología , Chile/epidemiología , Farmacorresistencia Bacteriana Múltiple/genética , Humanos , Infecciones por Klebsiella/epidemiología , Klebsiella pneumoniae/enzimología , Pruebas de Sensibilidad MicrobianaRESUMEN
Staphylococcus aureus isolates resistant to several antimicrobials have been gradually emerged since the beginning of the antibiotic era. Consequently, the first isolation of methicillin-resistant S. aureus occurred in 1960, which was described a few years later in Chile. Currently, S. aureus resistant to antistaphylococcal penicillins is endemic in Chilean hospitals and worldwide, being responsible for a high burden of morbidity and mortality. This resistance is mediated by the expression of a new transpeptidase, named PBP2a or PBP2', which possesses lower affinity for the ß-lactam antibiotics, allowing the synthesis of peptidoglycan even in presence of these antimicrobial agents. This new enzyme is encoded by the mecA gene, itself embedded in a chromosomal cassette displaying a genomic island structure, of which there are several types and subtypes. Methicillin resistance is mainly regulated by an induction mechanism activated in the presence of ß-lactams, through a membrane receptor and a repressor of the gene expression. Although mec-independent methicillin resistance mechanisms have been described, they are clearly infrequent.
Asunto(s)
Proteínas Bacterianas/genética , Estructuras Genéticas/genética , Staphylococcus aureus Resistente a Meticilina/genética , Proteínas de Unión a las Penicilinas/genética , Antibacterianos/química , Antibacterianos/farmacología , Proteínas Bacterianas/efectos de los fármacos , Cromosomas Bacterianos/efectos de los fármacos , Genes Bacterianos/efectos de los fármacos , Meticilina/química , Meticilina/farmacología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Estructura Molecular , Proteínas de Unión a las Penicilinas/efectos de los fármacosRESUMEN
Objective: To elucidate whether the genetic platforms of blaCTX-M contribute to the phenotypes of multi-drug-resistance (MDR) in the first carbapenemase-producing K. pneumoniae strains isolated in Chile. Method: Twenty-two carbapenemase-producing K. pneumoniae strains isolated from different Chilean patients and hospitals were studied. Their genetic relatedness was assessed by PFGE and MLST. The levels of antibiotic resistance were evaluated by determining the minimum inhibitory concentration of various antimicrobials. In addition, several antibiotic resistance genes of clinical relevance in Chile were investigated. The prevalence, allelic variants, and genetic platforms of blaCTX-M were determined by PCR and sequencing. Results: Out of the 22 strains studied, 20 carry KPC, one carries NDM-1, and one carries OXA-370. The PFGE analysis showed three clades with a genetic relatedness >85%, two formed by four strains and one by eight strains. The other strains are not genetically related, and a total of 17 different pulse types were detected. Ten different STs were identified, the main ones being ST258 (five strains) and ST1161 (seven strains). The isolates presented different percentages of resistance, and 82% were resistant to all the ß-lactams tested, 91% to ciprofloxacin, 73% to colistin, 59% to gentamicin, 50% to amikacin, and only 9% to tigecycline. All isolates carried blaTEM and blaSHV, whereas 71% carried aac(6')Ib-cr, and 57% one qnr gene (A, B, C, D, or S). The blaCTX-M gene was found in 10 of the isolates (4 blaCTX-M-15 and 6 blaCTX-M-2). The characterization of the platform, in seven selected strains, revealed that the gene is associated with unusual class 1 integrons and insertion sequences such as ISCR1, ISECp1, and IS26. Conclusion: In the first carbapenemase-producing K. pneumoniae strains isolated in Chile the genetic platform of blaCTX-M-2 corresponds to an unusual class 1 integron that can be responsible for the MDR phenotype, whereas the genetic platforms of blaCTX-M-15 are associated with different IS and do not contribute to multi-drug resistance.
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OBJECTIVE: To analyze the radiological aspects of pulmonary atelectasis in newborns on mechanical ventilation and treated in an intensive care unit, associating the characteristics of atelectasis with the positioning of the head and endotracheal tube seen on the chest X-ray, as well as with the clinical variables. MATERIALS AND METHODS: This was a retrospective cross-sectional study of 60 newborns treated between 1985 and 2015. Data were collected from medical records and radiology reports. To identify associations between variables, we used Fisher's exact test. The level of significance was set at p < 0.05. RESULTS: The clinical characteristics associated with improper positioning of the endotracheal tube were prematurity and a birth weight of less than 1000 g. Among the newborns evaluated, the most common comorbidity was hyaline membrane disease. Atelectasis was seen most frequently in the right upper lobe, although cases of total atelectasis were more common in the left lung. Malpositioning of the head showed a trend toward an association with atelectasis in the left upper lobe. CONCLUSION: Pulmonary atelectasis is a common complication in newborns on mechanical ventilation. Radiological evaluation of the endotracheal tube placement provides relevant information for the early correction of this condition.
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ABSTRACT: Molecular techniques that provide valuable information about the epidemiology of oral strains. The purpose of this study was to determine the genetic relatedness of 83 Enterococcus faecalis strains isolated from treated root canals. These strains were obtained from patients who were treated for persistent endodontic infections. E. faecalis isolates were molecular typed by Pulsed Field Gel Electrophoresis using Smal. Ten clonal groups and 13 pulse types with 38.7 % similarity for the less related strains were identified. Genetic heterogeneity among strains from different patients and a high level of genetic homogeneity among intrapatient strains were observed. Therefore, restriction endonuclease fingerprinting of genomic DNA from E. faecalis strains confirmed the polyclonality of the isolates obtained from the root canals of patients diagnosed with persistent endodontic infections, compared with other reports. These results provide additional data for a better understanding of the epidemiological aspects of root canal infections by E. faecalis.
RESUMEN: Las técnicas moleculares proporcionan información valiosa sobre la epidemiología de aislados orales. El propósito de este estudio fue determinar la relación genética de 83 cepas de Enterococcus faecalis aisladas de conductos radiculares tratados. Estas cepas se obtuvieron de pacientes que fueron tratados por infecciones endodónticas persistentes. Los aislados de E. faecalis se tipificaron molecularmente por electroforesis en gel de campo pulsado usando Smal. Se identificaron diez grupos clonales y 13 pulsotipos con un 38,7 % de similitud para las cepas menos relacionadas. Se observó heterogeneidad genética entre las cepas de diferentes pacientes y un alto nivel de homogeneidad genética entre las cepas intrapacientes. Por lo tanto, la toma de huellas dactilares a traves de restricción de ADN genómico de cepas de E. faecalis confirmó la policlonalidad de los aislados obtenidos de los conductos radiculares de pacientes diagnosticados con infecciones endodónticas persistentes, en comparación con otros informes. Estos resultados proporcionan datos adicionales para una mejor comprensión de los aspectos epidemiológicos de las infecciones del conducto radicular por E. faecalis.
Asunto(s)
Humanos , Periodontitis Periapical/microbiología , Enterococcus faecalis/aislamiento & purificación , Ápice del Diente/microbiología , ADN Bacteriano/análisis , Técnicas de Tipificación Bacteriana , Infecciones por Bacterias Grampositivas/microbiología , Electroforesis en Gel de Campo Pulsado , Cavidad Pulpar/microbiologíaRESUMEN
Abstract Objective: To analyze the radiological aspects of pulmonary atelectasis in newborns on mechanical ventilation and treated in an intensive care unit, associating the characteristics of atelectasis with the positioning of the head and endotracheal tube seen on the chest X-ray, as well as with the clinical variables. Materials and Methods: This was a retrospective cross-sectional study of 60 newborns treated between 1985 and 2015. Data were collected from medical records and radiology reports. To identify associations between variables, we used Fisher's exact test. The level of significance was set at p < 0.05. Results: The clinical characteristics associated with improper positioning of the endotracheal tube were prematurity and a birth weight of less than 1000 g. Among the newborns evaluated, the most common comorbidity was hyaline membrane disease. Atelectasis was seen most frequently in the right upper lobe, although cases of total atelectasis were more common in the left lung. Malpositioning of the head showed a trend toward an association with atelectasis in the left upper lobe. Conclusion: Pulmonary atelectasis is a common complication in newborns on mechanical ventilation. Radiological evaluation of the endotracheal tube placement provides relevant information for the early correction of this condition.
Resumo Objetivo: Analisar os aspectos radiológicos da atelectasia pulmonar em recém-nascidos com doenças clinicamente tratáveis submetidos a ventilação mecânica e atendidos em uma unidade de tratamento intensivo neonatal, associando características da atelectasia com o posicionamento da cabeça e da cânula endotraqueal na radiografia de tórax e com variáveis clínicas. Materiais e Métodos: Estudo de corte transversal e retrospectivo, em que foram incluídos 60 pacientes internados entre 1985 e 2015. A coleta dos dados foi realizada por meio da revisão dos prontuários e dos laudos das radiografias dos recém-nascidos. Para associação das variáveis foi realizado o teste exato de Fisher. O nível de significância adotado foi p < 0,05. Resultados: As características clínicas associadas à localização inadequada da cânula foram prematuridade e o peso ao nascer inferior a 1000 g. A doença clínica mais frequentemente encontrada foi a doença da membrana hialina. O lobo pulmonar superior direito foi o que apresentou maior frequência de atelectasia, e casos de atelectasia completa foram mais frequentes no pulmão esquerdo. O mau posicionamento da cabeça mostrou uma tendência de associação com atelectasia no lobo superior esquerdo. Conclusão: A atelectasia pulmonar representou uma complicação importante em recém-nascidos submetidos a ventilação mecânica, sendo a avaliação radiológica do posicionamento da cânula endotraqueal relevante para a correção precoce dessa condição.
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Malaria is a widespread disease caused mainly by the Plasmodium falciparum (Pf) and Plasmodium vivax (Pv) protozoan parasites. Depending on the parasite responsible for the infection, high morbidity and mortality can be triggered. To escape the host immune responses, Plasmodium parasites disturb the functionality of B cell subsets among other cell types. However, some antibodies elicited during a malaria infection have the potential to block pathogen invasion and dissemination into the host. Thus, the question remains, why is protection not developed and maintained after the primary parasite exposure? In this review, we discuss different aspects of B cell responses against Plasmodium antigens during malaria infection. Since most studies have focused on the quantification of serum antibody titers, those B cell responses have not been fully characterized. However, to secrete antibodies, a complex cellular response is set up, including not only the activation and differentiation of B cells into antibody-secreting cells, but also the participation of other cell subsets in the germinal center reactions. Therefore, a better understanding of how B cell subsets are stimulated during malaria infection will provide essential insights toward the design of potent interventions.
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Linfocitos B/inmunología , Malaria Falciparum/inmunología , Malaria Vivax/inmunología , Plasmodium falciparum/inmunología , Plasmodium vivax/inmunología , Anticuerpos Antiprotozoarios/inmunología , Antígenos de Protozoos/inmunología , Linfocitos B/parasitología , Interacciones Huésped-Parásitos/inmunología , Humanos , Inmunidad Celular/inmunología , Malaria Falciparum/parasitología , Malaria Vivax/parasitología , Plasmodium falciparum/fisiología , Plasmodium vivax/fisiologíaRESUMEN
OBJECTIVES: KPC-producing strains present a wide range of carbapenem minimum inhibitory concentrations (MICs). This variation may be due to differential expression of blaKPC and porin genes, efflux pump activity and the production of extended-spectrum ß-lactamases and/or AmpC ß-lactamases. The aim of this study was to determine the role of efflux pumps inhibited by phenylalanine-arginine ß-naphthylamide (PAßN) in resistance to carbapenems in Chilean clinical isolates of blaKPC-harbouring Klebsiella pneumoniae. METHODS: MICs were determined by the agar dilution method for imipenem, meropenem, ertapenem and ciprofloxacin in the presence and absence of PAßN (25mg/L) in 17 carbapenem-resistant KPC-producing K. pneumoniae strains. Outer protein membrane (OMP) profiles were determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Expression levels of the ompK35 and ompK36 genes were also determined by real-time quantitative reverse transcription PCR (qRT-PCR). RESULTS: No contribution of PAßN-inhibited efflux pumps to carbapenem resistance was found, unlike ciprofloxacin resistance. However, a ≥4-fold increase in the MIC of at least one carbapenem was observed in 13 isolates in the presence of PAßN. Additionally, decreased gene expression of ompK35 and ompK36 in the presence of PAßN was detected, however no obvious differences in porin band intensity were observed by SDS-PAGE. CONCLUSIONS: The presence of PAßN resulted in an increase in carbapenem MICs unrelated to efflux pump inhibition, and a decrease in the expression of ompK35 and ompK36 genes without an obvious difference in OMP profiles observed by SDS-PAGE. Therefore, additional factors are responsible for the increase in carbapenem MIC in the presence of PAßN.
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Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Carbapenémicos/farmacología , Dipéptidos/metabolismo , Inhibidores Enzimáticos/metabolismo , Klebsiella pneumoniae/enzimología , Proteínas de Transporte de Membrana/metabolismo , beta-Lactamasas/metabolismo , Proteínas de la Membrana Bacteriana Externa/análisis , Proteínas de la Membrana Bacteriana Externa/genética , Transporte Biológico Activo/efectos de los fármacos , Enterobacteriaceae Resistentes a los Carbapenémicos/efectos de los fármacos , Enterobacteriaceae Resistentes a los Carbapenémicos/enzimología , Enterobacteriaceae Resistentes a los Carbapenémicos/aislamiento & purificación , Chile , Electroforesis en Gel de Poliacrilamida , Perfilación de la Expresión Génica , Humanos , Infecciones por Klebsiella/microbiología , Klebsiella pneumoniae/química , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Resumen Desde el inicio de la era antimicrobiana se han ido seleccionando gradualmente cepas de Staphylococcus aureus resistentes a antimicrobianos de amplio uso clínico. Es así como en 1960 se describen en Inglaterra las primeras cepas resistentes a meticilina, y algunos años después son informadas en hospitales de Chile. Actualmente, S. aureus resistente a penicilinas antiestafilocóccicas es endémico en los hospitales de nuestro país y del mundo, siendo responsable de una alta morbimortalidad. La resistencia es mediada habitualmente por la síntesis de una nueva transpeptidasa, denominada PBP2a o PBP2' que posee menos afinidad por el β-lactámico, y es la que mantiene la síntesis de peptidoglicano en presencia del antimicrobiano. Esta nueva enzima se encuentra codificada en el gen mecA, a su vez inserto en un cassette cromosomal con estructura de isla genómica, de los cuales existen varios tipos y subtipos. La resistencia a meticilina se encuentra regulada, principalmente, por un mecanismo de inducción de la expresión del gen en presencia del β-lactámico, a través de un receptor de membrana y un represor de la expresión. Si bien se han descrito mecanismos generadores de resistencia a meticilina mec independientes, son categóricamente menos frecuentes.
Staphylococcus aureus isolates resistant to several antimicrobials have been gradually emerged since the beginning of the antibiotic era. Consequently, the first isolation of methicillin-resistant S. aureus occurred in 1960, which was described a few years later in Chile. Currently, S. aureus resistant to antistaphylococcal penicillins is endemic in Chilean hospitals and worldwide, being responsible for a high burden of morbidity and mortality. This resistance is mediated by the expression of a new transpeptidase, named PBP2a or PBP2', which possesses lower affinity for the β-lactam antibiotics, allowing the synthesis of peptidoglycan even in presence of these antimicrobial agents. This new enzyme is encoded by the mecA gene, itself embedded in a chromosomal cassette displaying a genomic island structure, of which there are several types and subtypes. Methicillin resistance is mainly regulated by an induction mechanism activated in the presence of β-lactams, through a membrane receptor and a repressor of the gene expression. Although mec-independent methicillin resistance mechanisms have been described, they are clearly infrequent.