TLR and NLRP3 inflammasome expression deregulation in macrophages of adult rats subjected to neonatal malnutrition and infected with methicillin-resistant Staphylococcus aureus.

OBJECTIVES: Nutritional aggression in critical periods may lead to epigenetic changes that affect gene expression. The aim of this study was to assess the effect of neonatal malnutrition on the expression of toll-like receptor (TLR)-2, TLR-4, and NLRP3 receptors, caspase-1 enzyme, and interleukin (IL)-1 ß production in macrophages infected with methicillin-resistant (MRSA) and methicillin-sensitive (MSSA) Staphylococcus aureus. METHODS: Wistar rats (N = 24) were divided in two distinct groups: nourished (17% casein) and malnourished (8% casein). Four systems were established after the isolation of mononuclear cells: negative control, positive control, MRSA, and MSSA. The plates were incubated at 37°C for 24 h in humidified atmosphere and 5% carbon dioxide. Tests were performed after this period to analyze the expression of standard recognition receptors, caspase-1 enzyme, and the production of IL-1 ß. Student's t test and analysis of variance were used in the statistical analysis; P < 0.05 was statistically significant. RESULTS: Malnutrition reduced animal growth and the expression of TLR-2, TLR-4, and NLRP3 receptors, the caspase-1 enzyme, and the IL-1 ß levels in macrophages infected with lipopolysaccharides in the present study. However, the interaction between the S. aureus and the macrophages promoted greater gene expression of receptors and enzymes. CONCLUSION: The neonatal malnutrition model compromised the expression of standard recognition receptors, of the caspase-1 enzyme as well as the production of IL-1 ß. However, the S. aureus and neonatal malnutrition combination led to intense transcription of such innate immunity components. Therefore, the deregulation in the expression of TLR and NLRP3 receptors and of the caspase-1 enzyme may induce extensive tissue injury and favor the permanence and spread of these bacteria, especially those that are methicillin resistant.

Clonal spread and accumulation of ß-lactam resistance determinants in Enterobacter aerogenes and Enterobacter cloacae complex isolates from infection and colonization in patients at a public hospital in Recife, Pernambuco, Brazil.

Enterobacter aerogenes and Enterobacter cloacae complex are the two species of this genus most involved in healthcare-associated infections that are ESBL and carbapenemase producers. This study characterized, phenotypically and genotypically, 51 isolates of E. aerogenes and E. cloacae complex originating from infection or colonization in patients admitted to a public hospital in Recife, Pernambuco, Brazil, by antimicrobial susceptibility profile, analysis of ß-lactamase genes (blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM, blaIMP and blaSPM), PCR and DNA sequencing, plasmid profile and ERIC-PCR. In both species, the genes blaTEM, blaCTX-M and blaKPC were detected. The DNA sequencing confirmed the variants blaTEM-1, blaCTX-M-15 and blaKPC-2 in isolates. More than one gene conferring resistance in the isolates, including the detection of the three previously cited genes in strains isolated from infection sites, was observed. The detection of blaCTX-M was more frequent in isolates from infection sites than from colonization. The gene blaKPC predominated in E. cloacae complex isolates obtained from infections; however, in E. aerogenes isolates, it predominated in samples obtained from colonization. A clonal relationship among all of E. aerogenes isolates was detected by ERIC-PCR. The majority of E. cloacae complex isolates presented the same ERIC-PCR pattern. Despite the clonal relation presented by the isolates using ERIC-PCR, different plasmid and resistance profiles and several resistance genes were observed. The clonal dissemination and the accumulation of ß-lactam resistance determinants presented by the isolates demonstrated the ability of E. aerogenes and E. cloacae complex, obtained from colonization and infection, to acquire and maintain different resistance genes.

IMMUNE RESPONSE OF SEVERE MALNUTRITION CHILDREN TREATED ACCORDING TO THE PROTOCOL OF THE WORLD HEALTH ORGANIZATION.

The aim of the study was to compare the innate immune system of severely malnourished children admitted to the Instituto de Medicina Integral Professor Fernando Figueira and treated according to the protocol of the World Health Organization (WHO) at admission and discharge. An experimental study was conducted with 20 children under two years of age. Ten of them had severe malnutrition and ten were a control group. The malnourished group consisted of hospitalized infants and it was submitted to WHO's protocol. Children with HIV and re-admitted during the study period were excluded. A blood sample was taken at admission and at discharge. Later, an analysis of blood leukocytes, adherence index, phagocytic capacity, production of free radicals superoxide and nitric oxide was performed. Patients with severe malnutrition at hospital discharge showed improved phagocytic function, release of oxygen radicals and reduction of the number of lymphocytes when compared to the time of admission. When compared to the control group, patients at hospital discharge had lower lymphocyte values and lower production of free radicals. Thus, it can be concluded that the duration of hospitalization was insufficient to restore cell-mediated immunity and microbicide activity.

El objetivo del estudio fue comparar el sistema inmune innato de niños con malnutrición grave ingresados en el Instituto de Medicina Integral Professor Fernando Figueira, tratados de acuerdo con el protocolo de la Organización Mundial de la Salud (OMS), al ingreso y al alta hospitalaria. Se llevó a cabo un estudio experimental con 20 niños menores de dos años de edad, 10 con malnutrición grave y 10 niños del grupo de control. El grupo de malnutridos se compuso de lactantes hospitalizados y sometidos al protocolo de la OMS. Se excluyeron los niños afectados por el HIV y los readmitidos durante el período del estudio. Se recogió una muestra de sangre al ingreso y otra al alta, y posterioriormente se realizó el análisis del perfil leucocitario, y el índice de adherencia, la capacidad fagocítica y la producción de los radicales libres superóxido y óxido nítrico. Los pacientes con malnutrición grave en el alta hospitalaria mostraron mejoría de la función fagocítica, la liberación de radicales oxidantes y la reducción del número de linfocitos en comparación con el ingreso hospitalario. En comparación con el grupo de control, los pacientes en el alta hospitalario presentaron valores más bajos de linfocitos y de producción de radicales libres. Por lo tanto, se puede concluir que el tiempo de hospitalización fue insuficiente para restablecer la inmunidad mediada por células, así como para restaurar la actividad microbicida.

Comparative study of bone regeneration in critical cranial bone defects using bone marrow adult stem cells and calcium phosphate / Estudio comparativo sobre regeneración ósea en defectos óseos craneales con células madre adultas de la médula ósea y fosfato cálcico (BoneSource(R))

Objective. To compare the use of Bone Marrow adult Stem Cells (BMSCs), differentiated in vitro into osteoblasts, associated to calcium phosphate versus autogenous bone graft, in the repair process of critical size bone defects. Materials and method. On 36 Wistar adult rats, bilateral full-thickness defects on parietal bone were created. The defects were either repaired with calcium phosphate (group I), calcium phosphate + (BMSCs) (group II) or autogenous bone graft (group III), and the opposite side with blood clot (Control Group). In all cases a collagen membrane was used. The animals were sacrificed at 30 and 60 days, and all specimens were collected for further histological and histomorfometric study. Results. At 30 days, group III (autogenous bone graft) evidences a statistical difference on bone formation when compared to the experimental and control groups (p ≤ 0.05). At 60 days group II (BS + BMSCs) and group III (autogenous bone) showed a similar bone formation and has only a statistical difference when compared to group I (BS) and control group. Conclusion. The use of calcium phosphate in conjunction with BMSCs resulted in a similar behavior in the process of bone repair in critical size defects, when compared with autogenous bone graft (AU)

Objetivo. Comparar el uso de células madre adultas de la médula ósea (CMMO), diferenciadas in vitro en osteoblastos, asociadas a fosfato cálcico, frente a injerto de hueso autólogo, en el proceso de reparación de defectos óseos de tamaño crítico. Material y Método. En 36 ratas adultas Wistar, se crearon defectos bilaterales de todo el grosor en el hueso parietal. Los defectos se repararon con fosfato de calcio (BoneSource®) (grupo I), fosfato de calcio (BoneSource®) + (CMMO) (grupo II) o injerto de hueso autólogo (grupo III), y en el lado contralateral con coágulo de sangre (grupo de control). En todos los casos se utilizó membrana de colágeno. Los animales fueron sacrificados a las 30 y 60 días y se obtuvieron todas las muestras para el estudio histológico y el análisis histomorfométrico. Resultados. A los 30 días, en el grupo III (injerto de hueso autólogo) se puso de manifiesto una diferencia estadísticamente significativa en la formación de hueso en comparación con el grupo experimental y el de control (p < 0,05). A los 60 días, en el grupo II (BoneSource®) + CMMO) y el grupo III (hueso autólogo) se demostró una formación ósea similar, y sólo se evidenció una diferencia estadísticamente significativa en comparación con el grupo I (BoneSource®) y el grupo de control. Conclusión. El uso de fosfato de calcio en combinación con CMMO indujo un comportamiento similar en el proceso de reparación ósea en defectos de tamaño crítico, en comparación con injerto de hueso autólogo (AU)

[Non-dermatophytic moulds: onychomycosis in four patients infected with the human immunodeficiency virus]. / Hongos filamentosos no dermatofitos: onicomicosis en cuatro pacientes infectados con el virus de la inmunodeficiencia humana.

Patients infected with human immunodeficiency virus (HIV) are a risk group for onychomycosis, fungal infections caused mainly by dermatophytes and yeast. However, non-dermatohytic moulds are becoming common agents for nail infections in this population of patients. We report four cases of onychomycosis caused by non-dermatophytic moulds (Aspergillus niger, Scytalidium hyalinum, Scytalidium dimidiatum and Fusarium solani) in patients infected with HIV from Recife, Pernambuco, Brazil. Onychomicosis by non-dermatophytic species in HIV-positive patients requires special attention in the clinical and the laboratory. A proper diagnosis is necessary to establish the specific and adequate treatment, preventing fungal invasion.

Hongos filamentosos no dermatofitos: onicomicosis en cuatro pacientes infectados con el virus de la inmunodeficiencia humana / Non-dermatophytic moulds: onychomychosis in four patients infected with the human immunodeficiency virus

La infección causada por el virus de la inmunodeficiencia humana (VIH) es unfactor de riesgo para el desarrollo de las onicomicosis, infecciones micóticascausadas principalmente por dermatofitos y levaduras. Los hongos filamentososno dermatofitos están emergiendo como agentes de lesiones ungueales eninmunodeprimidos. Presentamos cuatro casos de onicomicosis por hongosfilamentosos no dermatofitos (Aspergillus niger, Scytalidium hyalinum,Scytalidium dimidiatum y Fusarium solani) en pacientes portadores del VIH,residentes en la ciudad de Recife, Pernambuco - Brasil. Las onicomicosis porespecies no dermatofíticas en pacientes VIH-positivos requieren mayor atenciónen relación a los aspectos clínicos y de laboratorio con la finalidad deestablecer el diagnóstico etiológico correcto e indicar el tratamiento específico yadecuado, previniendo invasiones por hongos en otros sitios(AU)

Patients infected with human immunodeficiency virus (HIV) are a risk group foronychomycosis, fungal infections caused mainly by dermatophytes and yeast.However, non-dermatohytic moulds are becoming common agents for nailinfections in this population of patients. We report four cases of onychomycosiscaused by non-dermatophytic moulds (Aspergillus niger, Scytalidium hyalinum,Scytalidium dimidiatum and Fusarium solani) in patients infected with HIV fromRecife, Pernambuco, Brazil. Onychomicosis by non-dermatophytic species inHIV-positive patients requires special attention in the clinical and the laboratory.A proper diagnosis is necessary to establish the specific and adequatetreatment, preventing fungal invasion(AU)

Stress/aggressiveness-induced immune changes are altered in adult rats submitted to neonatal malnutrition.

BACKGROUND/AIMS: Neonatal malnutrition induces metabolic and endocrine changes that have beneficial effects on the neonatal in the short term but, in the longer term, these alterations lead to maladaptations. We investigated the effect of neonatal malnutrition on immune responses in adult rats submitted or not to an aggressiveness test. METHODS: Male Wistar rats were distributed to one of two groups according to their mothers' diet during lactation: the well-nourished group (group C, n = 42, receiving 23% of protein) and the malnourished group (group MN, n = 42, receiving 8% of protein). After weaning, all rats received normoproteic diet. Ninety days after birth, each group was subdivided into three subgroups: control rats (n = 14, respectively), aggressive rats (n = 14, respectively) and rats receiving foot shock (FS; n = 14, respectively). Plasma corticosterone concentration was measured after FS sessions. Leukocyte counts and humoral immunity were evaluated. RESULTS: In neonatal malnourished animals, FS-induced stress reduced plasma corticosterone concentration. Intraspecific aggressiveness induced alterations in leukocyte counts and antibody titers 7 and 15 days after immunization. Neonatal malnourished animals showed no changes in the immune parameters evaluated. CONCLUSIONS: Expression of intraspecific aggressiveness activates the immune system. Neonatal malnutrition seems to have a long-lasting effect on components of both neuroendocrine and immune functions.

[Evaluation of the effect of splenectomy with autologous spleen tissue implantation in some monocyte functions in children with hepatosplenic schistosomiasis mansoni]. / Avaliação do efeito da esplenectomia e auto-implante esplênico sobre algumas funções de monócitos em crianças com esquistossomose mansônica.

Adherence index, superoxide and TNF-alpha production in monocytes, with or without tuftsin treatment, were investigated in hepatosplenic schistosomiasis mansoni bearers with splenectomy with or without autologous implantation of spleen tissue. Three groups were evaluated: Healthy volunteers control group (CG) (n=12); Splenectomy with seft auto-transplant AG (n=18) and Splenectomy without auto-transplant WAG (n=9). Adherence index and TNF-alpha did not differ among the groups. Superoxide production was similar in CG and AG, in the 1st hour after cell stimulation. SP was larger in each hour time in CG and AG groups as compared WAS group. TT recovered normal pattern of SP in AG comparable with levels found in CG, with increase from the 1st to 2nd hour. However, TT did not alter SP in WAG, which remained reduced in all time points. Autologous implantation of spleen tissue seems to contribute for recovery and maintenance of the evaluated immunological reactions, which might be important in response to infections.

Avaliação do efeito da esplenectomia e auto-implante esplênico sobre algumas funções de monócitos em crianças com esquistossomose mansônica / Evaluation of the effect of splenectomy with autologous spleen tissue implantation in some monocyte functions in children with hepatosplenic schistosomiasis mansoni

Investigamos em portadores de esquistossomose hepatoesplênica após esplenectomia com ou sem auto-implante esplênico: índice de aderência, produção de superóxido (SP) e de TNF-alfa em monócitos, tratados ou não com tuftsina. Avaliamos três grupos: voluntários sadios CG (grupo controle) (n=12); esplenectomizados com auto-implante AG (n=18) e esplenectomizados sem auto-implante WAG (n=9). índice de aderência e TNF-alfa não diferiram entre os grupos. SP foi semelhante em CG e AG na 1ª hora após estimulação celular. SP foi maior em todos intervalos de tempo nos grupos CG e AG, comparados ao WAG. O tratamento com tuftsina recuperou o padrão de normalidade de SP em AG, com aumento da 1ª para a 2ª hora nos níveis do CG. O tratamento com tuftsina não alterou SP em WAG, permanecendo reduzida em todos intervalos. O auto-implante esplênico parece recuperar e manter os parâmetros imunológicos avaliados, que têm participação importante na resposta do hospedeiro às infecções.