RESUMO
Transient angiotensin-converting enzyme (ACE) inhibition induces persistent changes that protect against future nitric oxide synthase (NOS) inhibitor-induced cardiac fibrosis and inflammation. Given the role of fibroblasts in mediating these effects, the present study investigates whether prior ACE inhibition produced persistent changes in cardiac fibroblast physiology. Adult male spontaneously hypertensive rats (SHRs) were treated with vehicle (C+L) or the ACE inhibitor, enalapril (E+L) for 2 wk followed by a 2-wk washout period and a subsequent 7-day challenge with the NOS inhibitor N(ω)-nitro-l-arginine methyl ester. A third set of untreated SHRs served as controls. At the end of the study period, cardiac fibroblasts were isolated from control, C+L, and E+L left ventricles to assess proliferation rate, collagen expression, and chemokine release in vitro. After 7 days of NOS inhibition, there were areas of myocardial injury but no significant change in collagen deposition in E+L and C+L hearts in vivo. In vitro, cardiac fibroblasts isolated from C+L but not E+L hearts were hyperproliferative, demonstrated increased collagen type I gene expression, and an elevated secretion of the macrophage-recruiting chemokines monocyte chemoattractant protein-1 and granulocyte macrophage-colony stimulating factor. These findings demonstrate that in vivo N(ω)-nitro-l-arginine methyl ester treatment produces phenotypic changes in fibroblasts that persist in vitro. Moreover, this is the first demonstration that transient ACE inhibition can produce a persistent modification of the cardiac fibroblast phenotype to one that is less inflammatory and fibrogenic. It may be that the cardioprotective effects of ACE inhibition are related in part to beneficial changes in cardiac fibroblast physiology.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Cardiomiopatias/prevenção & controle , Enalapril/farmacologia , Fibroblastos/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Hipertensão/tratamento farmacológico , Animais , Cardiomiopatias/enzimologia , Cardiomiopatias/genética , Cardiomiopatias/patologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Quimiocina CCL2/metabolismo , Colágeno Tipo I/metabolismo , Modelos Animais de Doenças , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibrose , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Ventrículos do Coração/enzimologia , Ventrículos do Coração/patologia , Hipertensão/enzimologia , Hipertensão/genética , Hipertensão/patologia , Mediadores da Inflamação/metabolismo , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/metabolismo , Fenótipo , Ratos Endogâmicos SHR , Fatores de TempoRESUMO
BACKGROUND: Hemifacial spasm (HFS) is a chronic facial nerve disorder characterized by spontaneous muscle contractions. Microvascular decompression (MVD) is the neurosurgical treatment of choice. Intraoperative neurophysiologic monitoring (IOM) during MVD can help determine when adequate decompression is performed. METHODS: MVD with IOM was performed on 16 patients with HFS that included recording the abnormal lateral spread response (LSR) in lower facial muscles, considered as neurophysiologic marker of HFS. Two lower facial muscles were monitored as opposed to a standard monitoring of a single muscle. RESULTS: All patients underwent preoperative thin cut MRI confirming the presence of neurovascular conflict. Patients underwent small retrosigmoid craniotomy and MVD. In 13 cases, the LSR guided the surgeon to continue MVD until the response was unobtainable from all recorded lower facial muscles. In four of those (30%), the LSR persisted on one of the recorded muscle and prompted further exploration and decompression until complete disappearance of LSR in all recorded muscles. In two cases, the LSR disappeared after dural opening and never recurred during the procedure, therefore the completion of MVD was based on non reappearance of LSR. In one case, the LSR persisted despite apparent complete decompression of the nerve. Fourteen patients had complete relief of their symptoms after surgery, one had partial improvement and the one with persistent LSR was unchanged. CONCLUSION: Evaluation of the LSR by monitoring of two lower facial muscles provides valuable neurosurgical guidance during MVD for HFS. This simple modification of intra-operative monitoring may improve prediction of satisfactory MVD and HFS resolution.
Assuntos
Músculos Faciais/cirurgia , Espasmo Hemifacial/cirurgia , Cirurgia de Descompressão Microvascular , Monitorização Intraoperatória , Procedimentos Neurocirúrgicos , Feminino , Humanos , Masculino , Microcirurgia/métodos , Cirurgia de Descompressão Microvascular/métodos , Resultado do TratamentoRESUMO
OBJECTIVE: To evaluate the impact of a theory-based health education intervention on awareness of prostate cancer and intention to screen among men in Western Jamaica. METHODS: One hundred and eighty-eight men attending outpatient clinics in a hospital in Western Jamaica completed an interviewer-administered pretest survey. Following the pretest, participants received a health education intervention related to prostate cancer and an immediate post-test survey RESULTS: There were statistically significant increases in the percentage of correct responses between the pretest and post-test (p < 0.05). The greatest improvement was among items measuring knowledge of prostate cancer screening tests. Participants moved across the Stages of Change theoretical constructs indicating intention to screen. CONCLUSION: The sample was receptive to information about prostate cancer and the use of a theory-based educational intervention positively influenced knowledge of prostate cancer risk factors, symptoms, and types of screenings. PRACTICE IMPLICATIONS: This theory-based patient education programme can be replicated to promote awareness of prostate cancer and informed screening methods including potential risk associated with screening behaviours.
Assuntos
Educação em Saúde , Conhecimentos, Atitudes e Prática em Saúde , Neoplasias da Próstata/psicologia , Adulto , Idoso , Detecção Precoce de Câncer , Humanos , Jamaica , Masculino , Pessoa de Meia-Idade , Modelos Teóricos , Avaliação de Programas e Projetos de Saúde , Neoplasias da Próstata/diagnósticoAssuntos
Agonistas alfa-Adrenérgicos/farmacocinética , Efedrina/farmacocinética , Lactação/efeitos dos fármacos , Leite Humano/metabolismo , Agonistas alfa-Adrenérgicos/metabolismo , Adulto , Mama/irrigação sanguínea , Cromatografia Líquida de Alta Pressão , Efedrina/metabolismo , Feminino , Humanos , Lactente , Recém-Nascido , Lactação/metabolismo , Leite Humano/química , Prolactina/sangue , Ultrassonografia MamáriaRESUMO
An increase in the level of the tumor suppressor protein p53 can induce cell cycle arrest or cell death. Although mechanisms for regulating the life span of p53 have been described, there is growing evidence that transcriptional regulation of the p53 gene contributes significantly to controlling p53 protein levels and therefore the fate of a cell. However, the signal transduction pathways that lead to transcriptional activation of the p53 gene are poorly understood. The oncoprotein v-Maf and its cellular counterparts belong to the large combinatorially complex basic leucine zipper family of transcription factors, which include the AP1 family. To date few cellular targets of c-Maf have been identified. It is demonstrated here that v-Maf can bind as a homodimer to a variant Maf recognition element located between -66 and -54 upstream in the mouse p53 promoter. V-Maf and its cellular counterparts are shown to activate p53 expression through this site. The ability of v-Maf to activate p53 expression is modulated by AP1 family members. In addition, overexpression of v-Maf in primary cells leads to a p53-dependent cell death. Thus, Maf and members of the AP1 family are able to regulate p53 expression through this site in the p53 promoter.
Assuntos
Apoptose , Proteínas de Bactérias/metabolismo , Regiões Promotoras Genéticas , Ativação Transcricional , Proteína Supressora de Tumor p53/genética , Animais , Sequência de Bases , Galinhas , Sequência Consenso , DNA/metabolismo , Células HeLa , Humanos , Camundongos , Dados de Sequência MolecularRESUMO
Originally assumed to be antiviral substances, the efficacy of interferons in a number of pathologies, including malignancies, multiple sclerosis, and other immune syndromes, is increasingly recognized. This study provides data on the transfer of interferon alfa (2B) into human milk of a patient receiving massive intravenous doses for the treatment of malignant melanoma. Following an intravenous dose of 30 million IU, the amount of interferon transferred into human milk was only slightly elevated (1551 IU/mL) when compared to control milk (1249 IU/mL). These data suggest that even following enormous doses, interferon is probably too large in molecular weight to transfer into human milk in clinically relevant amounts.
Assuntos
Antineoplásicos/farmacocinética , Interferon-alfa/farmacocinética , Melanoma/tratamento farmacológico , Leite Humano/química , Neoplasias Cutâneas/tratamento farmacológico , Adulto , Feminino , HumanosRESUMO
Expression of the tumor suppressor protein p53 plays an important role in regulating the cellular response to DNA damage. During adenovirus infection, levels of p53 protein also increase. It has been shown that this increase is due not only to increased stability of the p53 protein but to the transcriptional activation of the p53 gene during infection. We demonstrate here that the E1a proteins of adenovirus are responsible for activating the mouse p53 gene and that both major E1a proteins, 243R and 289R, are required for complete activation. E1a brings about the binding of two cellular transcription factors to the mouse p53 promoter. One of these, ETF, binds to three upstream sites in the p53 promoter and one downstream site, whereas E2F binds to one upstream site in the presence of E1a. Our studies indicate that E2F binding is not essential for activation of the p53 promoter but that ETF is. Our data indicate the ETF site located downstream of the start site of transcription is the key site in conferring E1a responsiveness on the p53 promoter.
Assuntos
Proteínas E1A de Adenovirus/fisiologia , Proteínas de Transporte , Proteínas de Ciclo Celular , Proteínas de Ligação a DNA , Genes p53 , Fatores de Transcrição/metabolismo , Ativação Transcricional , Animais , Sítios de Ligação , Fatores de Transcrição E2F , Células HeLa , Humanos , Camundongos , Regiões Promotoras Genéticas , Ratos , Proteína 1 de Ligação ao Retinoblastoma , Fator de Transcrição DP1RESUMO
OBJECTIVE: Our aim was to determine the prevalence of associated vulvar adenocarcinoma, invasive Paget's disease, and recurrence of Paget's disease of the vulva. STUDY DESIGN: A retrospective review of tumor and pathology registries at 8 institutions is presented. Patients with recurrent disease were excluded. Histologic slide review was performed. RESULTS: The median age of the 100 patients was 70 years. The median duration of pruritus before surgery was 2 years. Thirty-four percent of patients experienced a recurrence at a median of 3 years. There was a 12% prevalence of invasive vulvar Paget's disease and a 4% prevalence of associated vulvar adenocarcinoma. One patient died of Paget's disease with associated vulvar adenocarcinoma. CONCLUSIONS: Paget's disease of the vulva is rarely associated with an underlying vulvar adenocarcinoma or invasive Paget's disease, but there is a high recurrence rate.
Assuntos
Adenocarcinoma/complicações , Adenocarcinoma/cirurgia , Doença de Paget Extramamária/complicações , Doença de Paget Extramamária/cirurgia , Neoplasias Vulvares/complicações , Neoplasias Vulvares/cirurgia , Adenocarcinoma/patologia , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Recidiva Local de Neoplasia/patologia , Doença de Paget Extramamária/patologia , Sistema de Registros , Estudos Retrospectivos , Neoplasias Vulvares/patologiaRESUMO
Epidemiological evidence has suggested that the declining prevalence of duodenal ulcer disease may be attributable to rising consumption of polyunsaturated fatty acids, a hypothesis supported by in vitro evidence of toxicity of such substances to Helicobacter pylori. The objective of the present study was to establish whether this association is causal. Forty patients with proven infection with H. pylori and endoscopic evidence of past or present duodenal ulcer disease were randomized to receive either polyunsaturated fatty acids (PUFA group), in the form of capsules and margarine, or a placebo (control). Both groups received concurrent H2 antagonist therapy. Efficacy of therapy was determined endoscopically by assessment of ulcer healing while H. pylori status was determined by antral biopsy, urease (EC 3.5.1.5) culture and histological assessment of the severity of H. pylori infection. Antral levels of prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) were quantified. Compliance was monitored. Before treatment, both groups were comparable for severity of H. pylori infection, smoking status and levels of LTB4 and PGE2. Despite a significant difference in consumption of linoleic acid (19.9 (SE 1.6) g for PUFA group v. 6.7 (SE 0.8) g for controls (P < 0.01) and linolenic acid (2.6 (SE 0.2) g v. 0.6 (SE 0.03) g (P < 0.01) there was no significant change in either the severity of H. pylori infection or prostaglandin levels in either group at 6 weeks. Consumption of a considerable amount of PUFA does not inhibit the colonization of the stomach by H. pylori nor does this alter the inflammatory changes characteristic of H. pylori gastritis. We conclude that the association between duodenal ulceration and a low level of dietary PUFA is likely to be spurious, probably reflecting the effect of confounding factors such as affluence, social class or smoking.
Assuntos
Úlcera Duodenal/terapia , Ácidos Graxos Insaturados/administração & dosagem , Infecções por Helicobacter/terapia , Helicobacter pylori , Terapia Combinada , Dinoprostona/metabolismo , Método Duplo-Cego , Úlcera Duodenal/metabolismo , Úlcera Duodenal/microbiologia , Feminino , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/metabolismo , Antagonistas dos Receptores H2 da Histamina/uso terapêutico , Humanos , Leucotrieno B4/metabolismo , Ácidos Linoleicos/administração & dosagem , Ácidos Linolênicos/administração & dosagem , Masculino , Pessoa de Meia-Idade , Antro Pilórico/metabolismo , Ranitidina/uso terapêutico , Falha de TratamentoRESUMO
Recombinant filarial proteins are of interest as potentially protective immunogens for lymphatic filariasis. We have previously identified paramyosin, myosin, and a heat shock protein 70 (HSP) 70 as prominent immunogens in individuals residing in an area endemic for lymphatic filariasis. Our goal in the present work was to identify the Brugia malayi tissues that contain these proteins. Polyclonal rabbit antisera with high levels of immunoglobulins to each of these proteins were prepared for use in indirect immunofluorescence microscopy studies of third-and fourth-stage larvae (L3's and L4's) and adult worms. Myosin and paramyosin were found within the longitudinal somatic musculature in all of these life stages. In L4's and adult worms, myosin and paramyosin were also detected within the walls of the reproductive and alimentary tracts of male and female worms. HSP 70 was evident within the somatic musculature, hypodermis, lateral chords, alimentary tract, and reproductive structures in L4's and adult worms. HSP 70 was not detected in sections of freshly obtained L3's. However, L3's cultured at 37 C for 24 hr before fixation demonstrated a classic heat shock response. In these larvae, intracellular HSP 70 was observed in all tissues. None of the antigens studied appeared to be located on cuticular surfaces.
Assuntos
Brugia Malayi/química , Proteínas de Choque Térmico HSP70/análise , Miosinas/análise , Tropomiosina/análise , Animais , Western Blotting , Reações Cruzadas , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Proteínas de Choque Térmico HSP70/imunologia , Humanos , Soros Imunes/imunologia , Larva/química , Masculino , Miosinas/imunologia , Coelhos , Tropomiosina/imunologiaRESUMO
A murine pulmonary model was used to study the mucosal immune response to Shigella flexneri serotype 2a infection. Inoculation of BALB/cJ mice with shigellae via the intranasal route resulted in bacterial invasion of bronchial and alveolar epithelia with concomitant development of acute suppurative bronchiolitis and subsequent development of lethal pneumonia. The pathology of pulmonary lesions resembled the colitis that characterizes shigellosis in humans and primates. Significant protection against a lethal dose of S. flexneri 2a was observed in mice previously infected with two sublethal doses of the homologous strain. Immunity against lethal challenge was associated with decreased bacterial invasion of the mucosal epithelium. Over the course of two sublethal challenges, which constituted primary and secondary immunizations, mice developed pulmonary and serum immunoglobulin G and A antibody recognizing both lipopolysaccharide and invasion plasmid antigens IpaB and IpaC. Immune mice and naive control mice differed in lung lavage cytokine levels following lethal challenge. Immune mice developed significantly elevated levels of pulmonary gamma interferon within 6 h of challenge, while naive control mice developed elevated levels of this cytokine later during the initial 24-h period. Both groups had elevated levels of gamma interferon during the 24- to 48-h period of infection. Both groups also had elevated levels of tumor necrosis factor alpha within 6 h of challenge, but the control mice had significantly higher levels at the 48- and 72-h time points. Elevated levels of interleukin-4 were observed only in immunized mice. This cytokine appeared within 24 h and receded between 48 and 72 h. Fluorescence-activated cell sorter analysis of lung parenchymal cells showed that both groups experienced an initial influx of monocytes, but the proportion of this cell type began to recede in immunized mice after 48 h of infection, while peak levels were maintained in the control animals. These studies suggest that elements of local B lymphocyte activity, as well as Th1 and Th2 lymphocyte activity, may contribute to the survival of immune mice after intranasal challenge with shigellae.
Assuntos
Anticorpos Antibacterianos/biossíntese , Citocinas/biossíntese , Disenteria Bacilar/imunologia , Pneumonia Bacteriana/imunologia , Shigella flexneri/imunologia , Administração Intranasal , Animais , Proteínas de Bactérias/imunologia , Bronquiolite , Modelos Animais de Doenças , Disenteria Bacilar/mortalidade , Disenteria Bacilar/prevenção & controle , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Imunização , Immunoblotting , Pulmão/imunologia , Pulmão/microbiologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Mucosa/imunologia , Pneumonia Bacteriana/mortalidade , Pneumonia Bacteriana/prevenção & controle , Sorotipagem , Shigella flexneri/classificação , Shigella flexneri/patogenicidade , Análise de SobrevidaRESUMO
We have investigated the transcription factor requirements for basal expression of the mouse p53 promoter by using a combination of reporter and electrophoretic mobility shift assays (EMSAs). We have found that only four regions of the promoter bind transcription factors in EMSAs, suggesting that these are the only important factors for basal transcription. These factors are NF1, USF, ETF-like and a novel factor which we have called PF2. Construction of promoter deletion mutants has shown that the absence of the PF2 site completely inactivates the promoter, whereas deletion of other sites, whilst reducing promoter activity, does not. We suggest that this novel transcription factor (PF2) is critical for expression of the mouse p53 promoter.
Assuntos
Proteínas de Transporte , Proteínas de Ciclo Celular , Regulação da Expressão Gênica , Genes p53 , Regiões Promotoras Genéticas , Fatores de Transcrição/metabolismo , Transcrição Gênica , Animais , Sequência de Bases , Linhagem Celular , DNA/genética , DNA/metabolismo , DNA Recombinante/metabolismo , Proteínas de Ligação a DNA/metabolismo , Fatores de Transcrição E2F , Genes Reporter , Humanos , Mamíferos , Camundongos , Dados de Sequência Molecular , Fatores de Transcrição NFI , Proteína 1 de Ligação ao Retinoblastoma , Deleção de Sequência , Especificidade da Espécie , Fator de Transcrição DP1 , Fatores Estimuladores UpstreamRESUMO
The p53 gene, which is frequently mutated in various tumors, encodes a phosphoprotein thought to have a key role in the regulation of cell proliferation. To explore their biological effects, the HeLa carcinoma line, which does not express p53, was co-transfected with plasmid constructs expressing wild-type or mutant p53 proteins, or unrelated proteins, along with a plasmid conferring resistance to a neomycin-kanamycin antibiotic analog (G418). Both wild-type and mutant forms of p53 stimulated the number of G418-resistant colonies between 5- and 36-fold. Further investigation of colony development revealed that p53 enhanced cell survival, leading to increased colony numbers, but did not stimulate cell growth. Nonetheless, we suggest that an initial slowing of cell growth caused by expression of the unintegrated p53 plasmids renders the transfectants resistant to selection with G418, thus causing a higher frequency of G418-resistant colonies. p53 constructs were found to be expressed transiently in HeLa cells as expected, but the G418-resistant colonies frequently failed to express p53. This loss of p53 expression may be due to negative regulatory effects of p53 on the cytomegalovirus promoter that drives the selection marker.
Assuntos
Genes p53 , Células HeLa/citologia , Proteína Supressora de Tumor p53/genética , Animais , Divisão Celular , Sobrevivência Celular , Expressão Gênica , Humanos , Técnicas In Vitro , Camundongos , RNA Mensageiro/genética , TransfecçãoRESUMO
A data base was compiled containing the amino acid sequences of 12 aspartate aminotransferases and 11 other aminotransferases. A comparison of these sequences by a standard alignment method confirmed the previously reported homology of all aspartate aminotransferases and Escherichia coli tyrosine aminotransferase. However, no significant similarity between these proteins and any of the other aminotransferases was detected. A more rigorous analysis, focusing on short sequence segments rather than the total polypeptide chain, revealed that rat tyrosine aminotransferase and Saccharomyces cerevisiae and Escherichia coli histidinol-phosphate aminotransferase share several homologous sequence segments with aspartate aminotransferases. For comparison of the complete sequences, a multiple sequence editor was developed to display the whole set of amino acid sequences in parallel on a single work-sheet. The editor allows gaps in individual sequences or a set of sequences to be introduced and thus facilitates their parallel analysis and alignment. Several clusters of invariant residues at corresponding positions in the amino acid sequences became evident, clearly establishing that the cytosolic and the mitochondrial isoenzyme of vertebrate aspartate aminotransferase, E. coli aspartate aminotransferase, rat and E. coli tyrosine aminotransferase, and S. cerevisiae and E. coli histidinol-phosphate aminotransferase are homologous proteins. Only 12 amino acid residues out of a total of about 400 proved to be invariant in all sequences compared; they are either involved in the binding of pyridoxal 5'-phosphate and the substrate, or appear to be essential for the conformation of the enzymes.
Assuntos
Aspartato Aminotransferases , Transaminases , Tirosina Transaminase , Sequência de Aminoácidos , Evolução Biológica , Dados de Sequência Molecular , Homologia de Sequência do Ácido NucleicoRESUMO
One of the chromosomal segments associated with virulence in Shigella flexneri encodes the production of aerobactin and the synthesis of an iron-regulated 76-kilodalton outer membrane protein believed to be the ferric-aerobactin receptor. However, S. flexneri expressing this putative aerobactin receptor, which is slightly larger than that encoded by pColV, is insensitive to the killing action of cloacin DF13, a bacteriocin which binds to other aerobactin receptor proteins and kills the cells. In this paper we show that the conjugal transfer of DNA encoding the iron-regulated 76-kilodalton protein from S. flexneri to Escherichia coli K-12 conferred cloacin DF13 sensitivity on the recipients. However, E. coli K-12 which had also inherited genes specifying Shigella O-antigen biosynthesis remained cloacin insensitive. The data suggest that it is unwise to use cloacin DF13 sensitivity alone to screen transconjugants or clinical isolates for the expression of aerobactin receptor proteins.
Assuntos
Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa/metabolismo , Bacteriocinas/farmacologia , Cloacina/farmacologia , Escherichia coli/genética , Ácidos Hidroxâmicos/fisiologia , Quelantes de Ferro/fisiologia , Shigella flexneri/efeitos dos fármacos , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Conjugação Genética , Resistência Microbiana a Medicamentos , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Ácidos Hidroxâmicos/metabolismo , Antígenos O , Receptores Imunológicos/genética , Shigella flexneri/genética , Shigella flexneri/imunologiaRESUMO
The serum antibody response to proteins encoded by the virulence-associated plasmid of Shigella flexneri was determined in monkeys challenged with virulent S. flexneri serotype 2a. With water-extractable antigen in an enzyme-linked immunosorbent assay, a significant increase in antibody titer against proteins from a plasmid-carrying, virulent strain of S. flexneri serotype 5 could be demonstrated in convalescent sera. There were minimal antibody titers against proteins from an avirulent (plasmid-free) organism. Previously identified plasmid-coded polypeptides a, b, c, and d were predominant antigens recognized by a majority of the convalescent sera in immunoblots. An additional 140-megadalton plasmid-coded polypeptide was also recognized by half of the sera. Convalescent serum from an infected monkey recognized antigens on the bacterial surface in several different plasmid-containing Shigella species and in an enteroinvasive Escherichia coli strain. A survey of sera obtained from children 5 to 10 years of age who had been infected with S. flexneri or S. sonnei revealed high enzyme-linked immunosorbent assay titers in both acute and convalescent sera against a water extract from a virulent Shigella strain. In contrast, children under 3 years of age had no antibody titer in either acute or convalescent sera against the virulence-associated shigella proteins, while 3- to 4-year-old children mounted an immune response against these proteins only in convalescence.
Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Disenteria Bacilar/imunologia , Shigella/imunologia , Animais , Anticorpos Antibacterianos/biossíntese , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina G/biossíntese , Macaca mulatta/imunologia , Peso Molecular , PlasmídeosRESUMO
Seven plasmid-coded polypeptides, designated a through g, were identified by two-dimensional nonequilibrium pH gradient electrophoresis of radiolabeled extracts from minicells of virulent Shigella flexneri serotypes 2a and 5 and enteroinvasive Escherichia coli O143. These polypeptides were deemed to be products of 140-megadalton (MDa) virulence-associated plasmids because they were not synthesized in minicells which were not harboring a 140-MDa plasmid or in minicells which were carrying an F lac plasmid of the same incompatibility group. Synthesis of these polypeptides was repressed in minicells incubated at 30 degrees C and in minicells isolated from a noninvasive opaque colonial variant, even though these strains harbored a 140-MDa plasmid. Enriched fractions of polypeptides b, c, and d were obtained from S. flexneri serotype 5 by preparative isoelectric focusing, and polyclonal rabbit antisera recognizing each polypeptide were raised. These antisera were able to detect cross-reacting plasmid-coded polypeptide antigens in S. flexneri serotype 3, Shigella sonnei, and enteroinvasive E. coli O143. In addition, Western blots of minicell extracts from S. flexneri serotype 5 or E. coli O143 indicated that plasmid-coded polypeptides a through d were recognized by convalescent antiserum from a monkey infected with S. flexneri serotype 2a.
Assuntos
Antígenos de Bactérias/isolamento & purificação , Proteínas de Bactérias/isolamento & purificação , Escherichia coli/análise , Plasmídeos , Shigella/análise , Animais , Proteínas de Bactérias/imunologia , Escherichia coli/genética , Escherichia coli/patogenicidade , Concentração de Íons de Hidrogênio , Soros Imunes/imunologia , Ponto Isoelétrico , Macaca mulatta , Fenótipo , Coelhos , Shigella/genética , Shigella/patogenicidade , VirulênciaRESUMO
Therapy of acute heavy metal poisoning is currently limited to a group of moderately toxic drugs containing sulfhydryl groups. N-Acetylcysteine (NAC) was used in these studies to determine if this sulfhydryl containing amino acid would reduce the overall mortality of a group of heavy metal compounds. D-Penicillamine and dimercaprol (BAL) were also used for comparison. Groups of at least 100 mice (28 g) were injected subcutaneously with 2-190 mg/kg of copper, arsenic, thallium or cadmium for LD50 determinations. Other groups were injected 30-60 min later with NAC (200 mg/kg), d-penicillamine (50 mg/kg), or BAL (10 mg/kg), and mortality was monitored for 2 weeks. The LD50 for each treatment group was determined by regression analysis of log-probit transformed data. In arsenite treatment group the survival time was lengthened in NAC-treated animals although the LD50 was not significantly changed. BAL was only slightly more effective than NAC. The mortality in animals given copper and treated with NAC was almost eliminated, except at the highest doses. BAL provided the greatest protection, whereas d-penicillamine produced the least. The LD50 of copper was significantly changed from 60.5 mg/kg in control groups to 139 mg/kg in NAC-treated groups, and to 150 mg/kg and 91 mg/kg in BAL and d-penicillamine-treated groups. NAC and BAL were totally ineffective in the treatment of thallium and cadmium poisoning.