RESUMO
PURPOSE: We investigated structural hypertrophy and functional hyperfiltration as compensatory adaptations after radical nephrectomy in patients with renal cell carcinoma according to the preoperative chronic kidney disease stage. MATERIALS AND METHODS: We retrospectively identified 543 patients who underwent radical nephrectomy for renal cell carcinoma between 1997 and 2012. Patients were classified according to preoperative glomerular filtration rate as no chronic kidney disease-glomerular filtration rate 90ml/min/1.73m2 or greater (230, 42.4%), chronic kidney disease stage II-glomerular filtration rate 60 to less than 90ml/min/1.73m2 (227, 41.8%), and chronic kidney disease stage III-glomerular filtration rate 30 to less than 60ml/min/1.73m2 (86, 15.8%). Computerized tomography performed within 2 months before surgery and 1 year after surgery was used to assess functional renal volume for measuring the degree of hypertrophy of the remnant kidney, and the preoperative and postoperative glomerular filtration rate per unit volume of functional renal volume was used to calculate the degree of hyperfiltration. RESULTS: Among all patients (mean age = 56.0y) mean preoperative glomerular filtration rate, functional renal volume, and glomerular filtration rate/functional renal volume were 83.2ml/min/1.73m2, 340.6cm3, and 0.25ml/min/1.73m2/cm3, respectively. The percent reduction in glomerular filtration rate was statistically significant according to chronic kidney disease stage (no chronic kidney disease 31.2% vs. stage II 26.5% vs. stage III 12.8%, P<0.001). However, the degree of hypertrophic functional renal volume in the remnant kidney was not statistically significant (no chronic kidney disease 18.5% vs. stage II 17.3% vs. stage III 16.5%, P = 0.250). The change in glomerular filtration rate/functional renal volume was statistically significant (no chronic kidney disease 18.5% vs. stage II 20.1% vs. stage III 45.9%, P<0.001). Factors that increased glomerular filtration rate/functional renal volume above the mean value were body mass index (P = 0.012), diabetes mellitus (P = 0.023), hypertension (P = 0.015), and chronic kidney disease stage (P<0.001). CONCLUSIONS: Patients with a lower preoperative glomerular filtration rate had a smaller reduction in postoperative renal function than those with a higher preoperative glomerular filtration rate due to greater degrees of functional hyperfiltration.
Assuntos
Carcinoma de Células Renais , Neoplasias Renais , Insuficiência Renal Crônica , Taxa de Filtração Glomerular , Humanos , Rim , Pessoa de Meia-Idade , Nefrectomia , Estudos RetrospectivosRESUMO
BACKGROUND: Vitamin B12 (cobalamin) deficiency can result in irreversible structural brain changes if not treated appropriately. Long-term use of acid-lowering agents (ALA) has been linked to vitamin B12 deficiency, but results are inconsistent. AIM: To evaluate the association between prolonged ALA use and vitamin B12 deficiency by performing a meta-analysis. METHODS: A systematic search was conducted using MEDLINE, PubMed, EMBASE, Current Contents, Cochrane Library, Google Scholar, Science Direct and Web of Science. Original data were abstracted from each study and used to calculate a pooled odds ratio and 95% confidence interval (95% CI). RESULTS: Of the articles reviewed, four case-control studies (4254 cases and 19,228 controls) and one observational study met full criteria for analysis. The long-term ALA use was significantly associated with development of vitamin B12 deficiency (hazard ratio 1.83, 95% CI: 1.36-2.46, P-value 0.00). CONCLUSION: Chronic use of ALA is a risk factor for developing vitamin B12 deficiency. Judicious prescribing of ALA and regular monitoring of vitamin B12 in patients who are inevitably on long-term ALA therapy are recommended.
Assuntos
Antiácidos/administração & dosagem , Antiácidos/efeitos adversos , Deficiência de Vitamina B 12/sangue , Deficiência de Vitamina B 12/induzido quimicamente , Estudos de Casos e Controles , Refluxo Gastroesofágico/sangue , Refluxo Gastroesofágico/tratamento farmacológico , Humanos , Deficiência de Vitamina B 12/diagnósticoRESUMO
MTB12 protein, also called CFP-2, is a major and early secreted component of Mycobacterium tuberculosis. However, its role during mycobacterial infection has been poorly characterized. In this study, we purified the native MTB12 protein and investigated the profile of MTB12-induced cytokines [interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-6], in early tuberculosis (TB) patients (n = 20) and healthy controls (n = 35). The cytokine profiles were compared with those induced by the 30-kDa antigen (Ag). In healthy controls, MTB12-induced IFN-gamma production was markedly decreased in peripheral blood mononuclear cells compared with 30-kDa Ag-induced IFN-gamma. In TB patients, the mean IFN-gamma level induced by MTB12 was lower than that induced by the 30-kDa Ag, albeit the difference was not significant. After 2 months of anti-TB therapy, both the MTB12- and 30-kDa-induced IFN-gamma levels were significantly increased in TB patients. MTB12-induced TNF-alpha and IL-6 levels were prominently upregulated in monocyte-derived macrophages from TB patients, but they were not significantly different from those induced by the 30-kDa Ag. Further, the activation of p38 mitogen-activated protein kinase and extracellular signal-regulated kinase was required for the induction of TNF-alpha and IL-6 by MTB12, as well as by the 30-kDa Ag. Collectively, these data suggest that the MTB12 protein plays an essential role for proinflammatory responses through the MAPK pathway during the early stages of human TB, even though its T-cell immunoreactivity is weaker than that of the 30-kDa Ag.
Assuntos
Antígenos de Bactérias/imunologia , Interferon gama/imunologia , Interleucina-6/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose/imunologia , Fator de Necrose Tumoral alfa/imunologia , Antígenos de Bactérias/isolamento & purificação , Antígenos de Bactérias/farmacologia , Antituberculosos/uso terapêutico , Western Blotting , Ensaio de Imunoadsorção Enzimática , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Humanos , Interferon gama/biossíntese , Interferon gama/metabolismo , Interleucina-6/biossíntese , Interleucina-6/metabolismo , Masculino , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Tuberculose/tratamento farmacológico , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Interleukin (IL)-12 and tumour necrosis factor (TNF)-alpha are both thought to be critical factors in the defence against mycobacteria but are known to play different roles. In this study, we investigated the regulatory pathways for IL-12 and TNF-alpha expression in human monocyte-derived macrophages (MDMs) after treatment with Mycobacterium tuberculosis H37Rv or the Triton X-100 solubilized proteins (TSP) purified from M. tuberculosis. We found a rapid phosphorylation of Akt and extracellular signal-regulated kinase (ERK), albeit with differential activation kinetics, in human MDMs treated with M. tuberculosis or TSP. Studies using inhibitors selective for phosphatidylinositol 3-kinase (PI 3-K) and ERK 1/2 show that both pathway plays an essential role in the induction of TNF-alpha at both the transcriptional and translational levels in human MDMs. In contrast, blockade of the PI 3-K/Akt or ERK 1/2 pathways significantly increased M. tuberculosis- or TSP-induced IL-12 p40 and p35 mRNA and bioactive p70 protein. The enhancement of IL-12 levels by inhibition of PI 3-K and ERK 1/2 was not reversed by neutralization of TNF-alpha or addition of rhTNF-alpha, suggesting that the negative regulation of IL-12 is not mediated by concomitant TNF-alpha suppression. Further, PI 3-K activity is required for the M. tuberculosis- or TSP-induced phosphorylation of ERK 1/2 activation. TSP from M. tuberculosis shows a similar dependency on the PI 3-K and ERK 1/2 pathways to those by M. tuberculosis. Collectively, these data suggest that the Th1-driving cytokine IL-12 and proinflammatory cytokine TNF-alpha are differentially regulated by PI 3-K and ERK 1/2 pathways in human MDMs during mycobacterial infection. These results may provide therapeutic targets for precise and specific fine-tuning of cytokine responses.
Assuntos
Interleucina-12/imunologia , Sistema de Sinalização das MAP Quinases , Fosfatidilinositol 3-Quinases/imunologia , Tuberculose/imunologia , Fator de Necrose Tumoral alfa/imunologia , Análise de Variância , Androstadienos/farmacologia , Antígenos de Bactérias/imunologia , Butadienos/farmacologia , Células Cultivadas , Cromonas/farmacologia , Ensaio de Imunoadsorção Enzimática , Flavonoides/farmacologia , Humanos , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Morfolinas/farmacologia , Mycobacterium tuberculosis/imunologia , Nitrilas/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Fosforilação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , WortmaninaRESUMO
Production of polyhydroxyalkanoates (PHAs) substituted with cyclohexyl groups by Pseudomonas oleovorans grown with 4-cyclohexylbutyric acid (4-CHB) and its mixtures with nonanoic acid (NA) was investigated. Addition of NA to medium gave rise to an increase in the total concentration of 3-hydroxy-4-cyclohexylbutyrate repeating unit in the PHAs, indicating that the bioconversion rate of 4-CHB to polyester was significantly improved by the cometabolic effect. Increasing the proportion of NA from 1.0 to 7.5 mM at a concentration of 10 mM total carbon substrate also accelerated the uptake speed of 4-CHB by the organism and resulted in an increase of the ratio of 3-hydroxynonanoate to 3-hydroxyheptanoate from 1.28 to 2.05. Differential scanning calorimetric analysis of the PHAs bearing the corresponding functional groups showed one melting transition and one glass transition temperature varying according to the composition. These results indicated that random copolyesters were obtained from the carbon substrates used in this study.
Assuntos
Ácidos Carboxílicos/metabolismo , Polímeros/química , Polímeros/metabolismo , Pseudomonas/metabolismo , Ácido Butírico/farmacologia , Carbono/química , Ácidos Carboxílicos/química , Ácidos Graxos/farmacologia , Pseudomonas/crescimento & desenvolvimento , Temperatura , Fatores de TempoRESUMO
The aim of this study was to determine whether the antibodies raised in burn patients by active immunization with a Pseudomonas aeruginosa OMPs vaccine have a protective efficacy against infection with P. aeruginosa. The binding patterns with P. aeruginosa OMPs of immunized burn patient sera were similar to the sera of immunized healthy humans as determined by immunoblot and immunoprecipitation analyses. The sera pooled from immunized burn patients after three immunizations showed a significantly higher opsonophagocytic-killing activity than the corresponding pre-immune sera, while the sera from unimmunized patients collected at the same day did not. Passive immunization of mice with post-immune sera of burn patients significantly enhanced the survival rate upon a lethal challenge with P. aeruginosa compared to the pre-immune sera, indicating the protective ability of the antibodies induced in burn patients by immunization. These results suggest that anti-P. aeruginosa OMPs antibodies elicited in burn patients by active immunization are protective against infection with P. aeruginosa, and provide a rational for further development of the vaccine for prevention against P. aeruginosa infection in burn patients.
Assuntos
Anticorpos Antibacterianos/biossíntese , Proteínas da Membrana Bacteriana Externa/uso terapêutico , Vacinas Bacterianas/uso terapêutico , Queimaduras/imunologia , Queimaduras/microbiologia , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa/imunologia , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/uso terapêutico , Reações Antígeno-Anticorpo , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Vacinas Bacterianas/imunologia , Humanos , Imunização Passiva/métodos , Imunoglobulina G/biossíntese , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Proteínas Opsonizantes/imunologia , Fagocitose/imunologia , Coelhos , Vacinas de Produtos Inativados/imunologia , Vacinas de Produtos Inativados/uso terapêuticoRESUMO
Outer membrane proteins (OMPs) of pathogenic bacteria have been used as protective antigens in developing bacterial vaccines. In the present study, we compared the antibody responses to a Pseudomonas aeruginosa OMP vaccine elicited in humans and rabbits by immunization. Immunization with the vaccine induced high titers of serum IgG antibody both in rabbits and humans but reactivities of the induced antibodies with the OMPs were different. The rabbit immune sera recognized most of the OMPs in the vaccine both in immunoblot and immunoprecipitation analyses. In contrast, a great variation in band pattern and intensity was observed among the human immune sera in immunoblot analysis, but not in immunoprecipitation analysis. Denaturation of the OMPs did not affect the binding activity of the rabbit immune sera as determined by ELISA, but substantially reduced those of the human immune sera and anti-OMP IgG purified from a pooled normal human plasma. These data suggest that antibody response to P. aeruginosa OMPs elicited by immunization in humans is mainly directed against discontinuous or conformation-dependent epitopes, which should be taken into account in developing vaccines, especially for OMP-derived synthetic peptides.
Assuntos
Anticorpos Antibacterianos/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Pseudomonas aeruginosa/imunologia , Animais , Anticorpos Antibacterianos/sangue , Proteínas da Membrana Bacteriana Externa/química , Vacinas Bacterianas/química , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Epitopos/química , Humanos , Immunoblotting , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Testes de Precipitina , Conformação Proteica , Desnaturação Proteica , Coelhos , VacinaçãoRESUMO
The aim of the present study was to compare two immunization schedules for a Pseudomonas aeruginosa outer membrane proteins (OMPs) vaccine in burn patients. In a double-blind, randomized and placebo-controlled clinical trial, 95 adult patients with burn injuries in 10% or greater of total body surface area were randomly allocated to either placebo or immunization groups. Three doses of the vaccine (0.5 or 1.0 mg) were administered intramuscularly at either 3- or 7-day intervals. The vaccine was well tolerated, and no severe adverse reactions were observed in any of the vaccinees. After three immunizations, 88 patients were available for evaluation of serum antibody titers. Elevation of OMPs-specific antibody titers in the immunization groups was significantly higher as compared with the placebo group, and the highest antibody response was obtained by immunization with 1.0-mg doses at 3-day intervals. Conventional blood culture, tissue culture of wound biopsy specimens and a nested polymerase chain reaction (PCR) assay of blood specimens were performed to determine the protective efficacy. The results of the nested PCR indicated that the overall detection rate of P. aeruginosa in blood was significantly lower among immunized patients than placebo patients (6.1 vs. 40.0%, P<0.001). Based on these results, we concluded that the P. aeruginosa OMPs vaccine is safe and highly immunogenic in burn patients, especially with 1.0-mg doses at 3-day intervals, and may be effective in conferring protection against P. aeruginosa bacteremia in burn patients.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Queimaduras/imunologia , Esquemas de Imunização , Pseudomonas aeruginosa/imunologia , Adulto , Idoso , Anticorpos Antibacterianos/sangue , Vacinas Bacterianas/efeitos adversos , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
In order to develop an effective means to treat Pseudomonas aeruginosa infections, we designed a large-scale process for purification of human IgG specific to P. aeruginosa outer membrane proteins (Oprs) from normal human sera. The process we developed includes affinity column chromatography using P. aeruginosa Oprs as ligands, protein A column chromatography and ultrafiltration, which enriched P. aeruginosa Oprs-specific IgG antibody by 500-fold. The purified anti-Oprs IgG was specific to the Oprs as confirmed by an ELISA competition assay and retained opsonophagocytic-killing capacity. In vivo protective efficacy of anti-Oprs IgG was evaluated by passive protection assays in mice where the 50% protective dose of anti-Oprs IgG against P. aeruginosa infections was 41 microg/kg, which was 20 times lower than that of normal serum IgG. When administered to mice 3 h after bacterial challenge, only anti-Oprs IgG afforded protection. These data demonstrate the feasibility of use of the purification process in producing functionally active target-specific human antibodies for clinical use and provide a rationale for use of anti-Oprs IgG as a valuable adjunct to treat P. aeruginosa infections.
Assuntos
Especificidade de Anticorpos , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/uso terapêutico , Imunoglobulina G/isolamento & purificação , Imunoglobulina G/uso terapêutico , Infecções por Pseudomonas/prevenção & controle , Pseudomonas aeruginosa/imunologia , Animais , Afinidade de Anticorpos , Vacinas Bacterianas/isolamento & purificação , Humanos , Imunoglobulina G/sangue , Masculino , Camundongos , Camundongos Endogâmicos ICR , Infecções por Pseudomonas/sangue , Infecções por Pseudomonas/imunologia , CoelhosRESUMO
In order to develop an effective means to treat and prevent Pseudomonas aeruginosa infections, we have purified P. aeruginosa outer membrane protein (Oprs)-specific human IgG antibody using a large-scale affinity column. In this study, we investigated the cross-protective activity of the purified anti-Oprs IgG against various immunotype strains of P. aeruginosa. The anti-Oprs IgG reacted with Oprs isolated from seven Fisher-Devlin immunotype strains of P. aeruginosa and was able to promote opsonophagocytic killing of all seven immunotype strains by human phagocytic cells. Administration of 500 microg anti-Oprs IgG to mice raised the LD50 of the P. aeruginosa strains by 8-250-fold, indicating the protective capacity against heterologous P. aeruginosa strains as well as homologous strains. In contrast, despite high titers against P. (aeruginosa Oprs, total serum IgG isolated from burn patient sera was no better than normal serum IgG in protecting mice from infection with P. aeruginosa. These data demonstrate that the affinity-purified human anti-Oprs IgG could afford protection against heterologous immunotype P. aeruginosa strains and provide a rationale to use anti-Oprs IgG as an adjunct for treatment of P. aeruginosa infections in humans.
Assuntos
Anticorpos Antibacterianos/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Imunoglobulina G/imunologia , Pseudomonas aeruginosa/imunologia , Animais , Reações Cruzadas , Modelos Animais de Doenças , Humanos , Imunização Passiva , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fagocitose/imunologia , Especificidade da EspécieRESUMO
In order to evaluate in humans the safety and immunogenicity of a Pseudomonas aeruginosa vaccine composed of outer membrane proteins (OMPs), CFC-101, we carried out a phase I/IIa clinical trial in healthy male volunteers. Groups of six volunteers were immunized either subcutaneously (s.c.) or intramuscularly (i.m.) with three dosages of the vaccine three times at 7-day intervals. The vaccine was well tolerated by volunteers. Local reactions in the injection sites were generally mild and transient. Significant increases in OMP-specific antibody were observed in both route groups after vaccinations but was higher in the i.m.-immunized group, where vaccination with 0.5 or 1.0 mg doses yielded 100% seroconversion. The specificity of the induced antibodies to P. aeruginosa OMP was demonstrated by western blot analysis and immunoprecipitation assay. An increase in Clq-binding capacity and ability to confer mice protection from lethal challenges with P. aeruginosa indicated the protective efficacy of the elicited antibodies. Based on these data, we concluded that the P. aeruginosa OMP vaccine is safe and effective in humans with an optimal dose of 0.5 and 1.0 mg and that i.m. is the better route than s.c. for this vaccine.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/farmacologia , Pseudomonas aeruginosa/imunologia , Adulto , Animais , Anticorpos Antibacterianos/sangue , Especificidade de Anticorpos , Vacinas Bacterianas/efeitos adversos , Vacinas Bacterianas/imunologia , Ativação do Complemento , Humanos , Imunização Passiva , Técnicas In Vitro , Leucócitos Mononucleares/imunologia , Masculino , Camundongos , Camundongos Endogâmicos ICR , Pessoa de Meia-Idade , Infecções por Pseudomonas/imunologia , Infecções por Pseudomonas/prevenção & controle , Segurança , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Malonate decarboxylase from Acinetobacter calcoaceticus was isolated and characterized (Kim, Y.S., Byun, H.S., J. Biol. Chem. 269 (1994) 29636-29641), and its subunits were reanalyzed recently to be alpha, beta, gamma, and delta. The genes for the subunits, MdcA (548 a.a.), B (295 a.a.), C (238 a.a.), and D (102 a.a.), of the enzyme have been cloned by using oligonucleotide primers deduced from amino acid sequences of peptides isolated from the purified enzyme, and sequenced to be clustered in an operon in the order of A-D-B-C. The operon was found to encode more genes than mdcABCD. The Escherichia coli, transformed with the vector containing the insert mdcADBC and about 1.7 kb of an upstream region, expressed the four subunits of the enzyme but the proteins did not show enzyme activity. It indicates that, like the enzymes from Malonomonas rubra and Klebsiella pneumoniae, more genes are needed for the formation of the functional malonate decarboxylase.