RESUMO
Serogroup B meningococcus (MenB) is a leading cause of meningitis and sepsis across the world and vaccination is the most effective way to protect against this disease. 4CMenB is a multi-component vaccine against MenB, which is now licensed for use in subjects >2 months of age in several countries. In this study, we describe the development and use of an ad hoc protein microarray to study the immune response induced by the three major 4CMenB antigenic components (fHbp, NHBA and NadA) in individual sera from vaccinated infants, adolescents and adults. The resulting 4CMenB protein antigen fingerprinting allowed the identification of specific human antibody repertoire correlating with the bactericidal response elicited in each subject. This work represents an example of epitope mapping of the immune response induced by a multicomponent vaccine in different age groups with the identification of protective signatures. It shows the high flexibility of this microarray based methodology in terms of high-throughput information and minimal volume of biological samples needed.
Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Infecções Meningocócicas/imunologia , Vacinas Meningocócicas/imunologia , Neisseria meningitidis Sorogrupo B/imunologia , Adolescente , Adulto , Anticorpos Antibacterianos/sangue , Criança , Pré-Escolar , Mapeamento de Epitopos , Humanos , Lactente , Infecções Meningocócicas/prevenção & controle , Biblioteca de Peptídeos , Análise Serial de Proteínas , Ensaios de Anticorpos Bactericidas Séricos , Adulto JovemRESUMO
Chronic hepatitis C virus (HCV) infection occurs in about 3 percent of the world's population and is a major cause of liver disease. HCV infection is also associated with cryoglobulinemia, a B lymphocyte proliferative disorder. Virus tropism is controversial, and the mechanisms of cell entry remain unknown. The HCV envelope protein E2 binds human CD81, a tetraspanin expressed on various cell types including hepatocytes and B lymphocytes. Binding of E2 was mapped to the major extracellular loop of CD81. Recombinant molecules containing this loop bound HCV and antibodies that neutralize HCV infection in vivo inhibited virus binding to CD81 in vitro.
Assuntos
Antígenos CD/metabolismo , Hepacivirus/metabolismo , Proteínas de Membrana/metabolismo , Proteínas do Envelope Viral/metabolismo , Sequência de Aminoácidos , Animais , Anticorpos/imunologia , Anticorpos Antivirais/imunologia , Antígenos CD/química , Antígenos CD/genética , Antígenos CD/imunologia , Linhagem Celular , DNA Complementar , Biblioteca Gênica , Hepacivirus/imunologia , Hepatite C/imunologia , Humanos , Fígado/citologia , Fígado/imunologia , Fígado/virologia , Linfócitos/imunologia , Linfócitos/virologia , Proteínas de Membrana/química , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Dados de Sequência Molecular , Pan troglodytes , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Alinhamento de Sequência , Tetraspanina 28 , Células Tumorais Cultivadas , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologiaRESUMO
The anti-inflammatory effects resulting from raising the levels of palmitoylethanolamide (PEA), an endogenous bioactive lipid, led to envisage N-Acylethanolamine Acid Amidase (NAAA), the cysteine hydrolase mainly responsible for PEA degradation, as an attractive target for small molecule inhibitors. Previous work in our group identified serine-derived ß-lactams as potent and systemically active inhibitors of NAAA activity. Aiming to expand the SAR study around this class of compounds, we investigated the effect of the substitution on the endocyclic nitrogen by designing and synthesizing a series of N-substituted ß-lactams. The present work describes the synthesis of new N-O-alkyl and N-O-aryl substituted ß-lactams and reports the results of the structure activity relationship (SAR) study leading to the discovery of a novel, single-digit nanomolar NAAA inhibitor (37). Compound 37 was shown in vitro to inhibit human NAAA via S-acylation of the catalytic cysteine, and to display very good selectivity vs. human Acid Ceramidase, a cysteine amidase structurally related to NAAA. Preliminary in vivo studies showed that compound 37, administered topically, reduced paw edema and heat hyperalgesia in a carrageenan-induced inflammation mouse model. The high in vitro potency of 37 as NAAA inhibitor, and its encouraging in vivo activity qualify this compound as a new tool for the study of the role of NAAA in inflammatory and pain states.
Assuntos
Amidoidrolases/antagonistas & inibidores , Inibidores Enzimáticos/síntese química , beta-Lactamas/farmacologia , Animais , Modelos Animais de Doenças , Desenho de Fármacos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Humanos , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Camundongos , Dor/tratamento farmacológico , Relação Estrutura-Atividade , beta-Lactamas/síntese química , beta-Lactamas/químicaRESUMO
To investigate the impairment of beta-adrenoceptor responsiveness in human hypertension, we evaluated the effect of an oral salt load (400 mEq/day of NaCl for 7 days) on plasma catecholamine concentrations and beta-adrenoceptor-mediated effects in 11 young patients with mild essential hypertension. Responses of heart rate and plasma cAMP to isoproterenol administration were used as indices of beta-adrenoceptor responsiveness. Salt loading induced a significant reduction in the dose of isoproterenol required to raise the heart rate by 25 bpm (CD25) (from 7.6 +/- 1.5 to 5.3 +/- 0.9 micrograms, p less than 0.05) and an increase in the slopes of the regression lines for heart rate changes and isoproterenol doses (delta HR/IS) (from 3.3 +/- 0.6 to 4.7 +/- 0.7, p less than 0.05) and for plasma cyclic AMP (cAMP) level changes and isoproterenol doses (delta cAMP/IS) (from 0.3 +/- 0.06 to 1.4 +/- 0.3, p less than 0.05). After salt loading there was a significant reduction in plasma catecholamine concentrations with a significant relationship between changes in upright plasma epinephrine levels and changes in CD25 (r = 0.904, p less than 0.01) and in the slopes for delta HR/IS (r = 0.983, p less than 0.001) and delta cAMP/IS (r = 0.922, p less than 0.001). These results support the hypothesis that the impairment of beta-adrenoceptor sensitivity observed in human hypertension is associated with a beta-adrenoceptor overstimulation due to chronically elevated adrenergic tone.
Assuntos
Hipertensão/metabolismo , Receptores Adrenérgicos beta/fisiologia , Receptores Adrenérgicos/fisiologia , Adolescente , Adulto , Aldosterona/sangue , Pressão Sanguínea/efeitos dos fármacos , AMP Cíclico/sangue , Epinefrina/sangue , Frequência Cardíaca/efeitos dos fármacos , Humanos , Hipertensão/fisiopatologia , Isoproterenol/farmacologia , Masculino , Norepinefrina/sangue , Receptores Adrenérgicos beta/efeitos dos fármacos , Renina/sangue , Cloreto de Sódio/farmacologiaRESUMO
Synthetic peptides corresponding to amino acid sequences of amino terminal non-alpha helical domains of human cytokeratin 18 and to low molecular weight human neurofilament subunit were used to obtain monospecific antisera. The results of our immunohistochemical investigations confirmed in general the data previously published on the distribution of cytokeratin 18 in human, rat, and calf tissues. The reactivity of the antiserum was abolished after formalin fixation of specimens. Immunolocalization of the neurofilament subunit using our monospecific antiserum was quite variable from species to species in cells of the central and peripheral nervous systems, and also varied as the result of the tissue fixation procedures. In particular, formalin fixation destroyed the immunoreactivity of the recognized epitope. We discuss the advantages and limits of the use of synthetic peptides as immunogens to produce polyclonal antibodies against intermediate filament proteins, with particular attention to the epitope masking phenomena in cytokeratin polypeptides and the phosphorylation of epitopes in neurofilament subunits.
Assuntos
Anticorpos Monoclonais/imunologia , Proteínas de Filamentos Intermediários/imunologia , Peptídeos/imunologia , Sequência de Aminoácidos , Aminoácidos/análise , Animais , Bovinos , Sistema Nervoso Central/análise , Sistema Nervoso Central/citologia , Sistema Nervoso Central/imunologia , Humanos , Immunoblotting , Imuno-Histoquímica/métodos , Proteínas de Filamentos Intermediários/análise , Queratinas/análise , Queratinas/imunologia , Dados de Sequência Molecular , Peptídeos/análise , RatosRESUMO
We have investigated the effects of 2',5'-dideoxyadenosine (DDA), 9'-beta-D-arabinofuranosyladenine (ARA), and 9-beta-D-xylofuranosyladenine (XFA), which have been classified as P-site adenosine agonists, on the cyclic adenosine 3',5'-monophosphate (cAMP) metabolism of human lymphocytes and polymorphonuclear leukocytes (PMNs). DDA (10(-5)-2 x 10(-4) M), ARA and XFA caused a dose-dependent decrease in cAMP content of human lymphocytes. In addition to decreasing lymphocyte cAMP levels, DDA, ARA, and XFA markedly inhibited the effects of many adenylate cyclase-stimulating agents including beta-adrenergic stimuli, prostaglandin E1 (PGE1), histamine, adenosine, forskolin and cholera toxin. Theophylline and 3-isobutyl-1-methylxanthine, which are known antagonists of adenosine A1/Ri and A2/Ra receptors, did not modify the inhibiting effects of DDA. Mn2+ (1 mM) increased the sensitivity to inhibition of adenylate cyclase agonists by DDA. We also search for the presence of adenosine P-sites in human PMNs. DDA caused a significant decrease of PMN cAMP levels only at the highest concentrations used (2 x 10(-4) M). In contrast, even low concentrations of DDA (10(-6)-10(-4) M) concentration-dependently blocked the stimulatory effect of PGE1 and forskolin on PMN cAMP accumulation. The results support the existence of a purine P-site that regulates cAMP metabolism of human lymphocytes and PMNs.
Assuntos
Adenosina/metabolismo , AMP Cíclico/metabolismo , Linfócitos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Receptores Purinérgicos/metabolismo , Adenosina/análogos & derivados , Adenosina/farmacologia , Adenilil Ciclases/metabolismo , Adulto , Células Cultivadas , Didesoxiadenosina/análogos & derivados , Didesoxiadenosina/farmacologia , Ativação Enzimática/efeitos dos fármacos , Humanos , Linfócitos/metabolismo , Neutrófilos/metabolismo , Antagonistas Purinérgicos , Receptores Purinérgicos/efeitos dos fármacos , Vidarabina/farmacologiaRESUMO
OBJECTIVES: a) To study the capacity of the technique of high-frequency color Doppler to detect flow signal of left internal mammary artery grafts; b) to assess the usefulness of an echo-enhancer agent to facilitate the detection of the signal, and c) to evaluate the patency of the graft according to its pulsed Doppler velocity profile pattern. METHODS: 39 consecutive patients were studied. A Hewlett-Packard 5500 echocardiograph was used, with a high-frequency probe (S12) applied at the high left parasternal border. When a graft signal was not elicited after a predetermined 5-minute check period, an intravenous dose of 4 g of Levovist (Schering España) at 400 mg/ml was administrated. According to previous studies, a pulsed Doppler flow profile with a predominantly diastolic pattern was considered a normal graft patency, while a systolic one was deemed as abnormal. RESULTS: Graft flow was identified by color Doppler in 33/39 patients (85%). The additional use of an echo-enhancer in 6 patients with no detected signal increased this proportion to 38/39 (97%). Normal flow patterns were seen in 34/38 (89%). Among the four patients with abnormal pattern, 1 case of early myocardial infarction was observed, while angiographic studies showed distal occlusion of the graft in 1 or the presence of competitive flow in 2 patients. CONCLUSIONS: The high-frequency color Doppler technique allows the detection of a flow signal from internal mammary artery grafts in most patients. The administration of an echo-enhancer agent is useful in those with non detectable signals. An abnormal pulsed Doppler velocity pattern indicates graft malfunction.
Assuntos
Meios de Contraste , Ecocardiografia Doppler em Cores , Anastomose de Artéria Torácica Interna-Coronária , Artéria Torácica Interna/diagnóstico por imagem , Polissacarídeos , Grau de Desobstrução Vascular , Velocidade do Fluxo Sanguíneo , Ecocardiografia Doppler em Cores/instrumentação , Ecocardiografia Doppler em Cores/métodos , Oclusão de Enxerto Vascular/diagnóstico , Humanos , Imageamento por Ressonância Magnética , Artéria Torácica Interna/patologia , Artéria Torácica Interna/fisiopatologia , Estudos ProspectivosRESUMO
This paper was presented at the 1st Update Course in Hysteroscopy and Microcolpohysteroscopy organised by LAMM and held at the 1st Institute of Obstetrics and Gynecology at Policlinico Umberto I in Rome. The paper is divided into three sections: the first section describes the technical instruments used during hysteroscopy (light source, means of distending the uterine cavity, optic systems); the second reports the technique used in hysteroscopic examination: in particular, the authors underline the importance where necessary (i.e. in young patients with stenosis of the cervical canal and in elderly patients with atrophy of the neck of the uterus or conglutination of the external uterine operning) of preceding dilatation of the cervical canal by the use of endovaginal prostaglandin derivatives in order to render it less traumatic. Moreover, the authors attempt to simplify the technique as much as possible so as to render it a routine or ambulatorial test; the third section describes complications of the hysteroscopic examination, in order of frequency, as well as methods of storing and disinfecting the instruments used in hysteroscopy.
Assuntos
Histeroscópios , Feminino , Doenças dos Genitais Femininos/diagnóstico , Humanos , Histeroscopia/métodos , ItáliaRESUMO
The Authors report the results of their research into the use of the alpha-interferon in the microcondylomatosis of the female genital apparatus. The therapy was successful in 18% of cases. Considering oncological risk connected with the permanency of the HPV in the uterine portio, the Authors consider they must continue the study on new diagnostic and therapeutic protocols.
Assuntos
Condiloma Acuminado/terapia , Doenças dos Genitais Femininos/terapia , Interferon-alfa/uso terapêutico , Estudos de Avaliação como Assunto , Feminino , HumanosAssuntos
Adenosina/metabolismo , Inflamação/fisiopatologia , Linfócitos/fisiologia , Receptores de Superfície Celular/metabolismo , Alprostadil , AMP Cíclico/metabolismo , Humanos , Cinética , Linfócitos/efeitos dos fármacos , Prostaglandinas E/farmacologia , Receptores Purinérgicos , Teofilina/farmacologiaRESUMO
B. Subtilis expression plasmids generally require a stringent Shine-Dalgarno Sequence (SDS). Site-directed-mutagenesis was explored to change the Shine-Dalgarno Sequence from AAAAATGGGG (mutant type) to AAAAAGGGGG (wild type) in recombinant plasmid PSM604. The single base substitution made the plasmid with wild SDS unstable in structure and segregation. The interaction of SDS with subtilisin leader sequence of PSM604 might be responsible for the instability of plasmid.
Assuntos
Bacillus subtilis/genética , Plasmídeos/genética , Sequência de Bases , Expressão Gênica , Mutagênese Sítio-Dirigida , Recombinação GenéticaRESUMO
Three diphtheria toxin (DT) mutants CRM-197, DT-del (148) and DT-E148S-K516A-F530A were cloned in B. Subtilis plasmid PSM604 under the subtilisin signal sequence. The expression was effective in both SMS300 and SMS118, but higher yield of 7.1 mg/L was observed in SMS300 compared with 2.1 mg/L in SMS118. Western blot showed that the recombinant protein could be effectively secreted into the culture medium as a 58 ku peptide, and could be degraded into two peptides of 37 ku and 21 ku.
Assuntos
Proteínas de Bactérias , Toxina Diftérica , Bacillus subtilis/genética , Toxina Diftérica/biossíntese , Plasmídeos , Proteínas Recombinantes/genéticaRESUMO
Adenosine is a natural nucleoside that plays a physiological role in the modulation of human inflammatory cells. We have investigated the presence of adenosine A2/Ra and A1/Ri receptors and of the P-site on human inflammatory cells. Human B and T(OKT4+ and OKT8+) lymphocytes, polymorphonuclear leucocytes, monocytes, basophils and platelets possess a membrane adenosine A2/Ra receptor. The activation of adenosine A2/Ra receptor increases the intracellular level of cyclic AMP in these cells. Human lymphocytes and neutrophils possess also an inhibitory adenosine A1/Ri receptor and a P-site whose activation inhibits the effect of many adenylate cyclase agonists including isoproterenol, PGE1, histamine, adenosine, cholera toxin and forskolin.
Assuntos
Adenosina/fisiologia , Leucócitos/metabolismo , Receptores de Superfície Celular/fisiologia , Adenosina/análogos & derivados , Adenosina/farmacologia , Adenilil Ciclases/metabolismo , AMP Cíclico/metabolismo , Humanos , Leucócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Receptores de Superfície Celular/efeitos dos fármacos , Receptores PurinérgicosRESUMO
Filaggrin is a protein normally present in the granular and horny layer of stratified squamous epithelia. We studied the presence of this protein in 83 benign lesions and in 73 cases of malignant epithelial tumours of the oral cavity and investigated its possible role as an immunohistochemical marker. The immunohistochemical technique was based on the P.A.P. method. The results in benign lesions show a distribution of filaggrin similar to that observed in the normal mucosa. By contrast, an irregular distribution of filaggrin is observed in areas of leukoplakia with parakeratosis and in papillomas. In malignant lesions the expression of this protein is closely related to the degree of differentiation of the cellular elements, being positive in more differentiated and negative in anaplastic areas. Therefore in some types of benign lesions filaggrin testifies an alteration of the normal process of keratinization. Filaggrin is more significant in malignant lesions in which its presence if any permits an evaluation of the degree of differentiation of the tumour.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Proteínas de Filamentos Intermediários/metabolismo , Leucoplasia/metabolismo , Mucosa Bucal/patologia , Neoplasias Bucais/metabolismo , Neoplasias Palatinas/metabolismo , Carcinoma de Células Escamosas/patologia , Proteínas Filagrinas , Humanos , Imuno-Histoquímica , Leucoplasia/patologia , Neoplasias Bucais/patologia , Neoplasias Palatinas/patologiaRESUMO
Normal bladder urothelium and large spectrum bladder lesions have been investigated by immunohistochemistry with monoclonal antibodies of variable specificity (SK 56-23, a large spectrum antibody; SK 60-61, which reacts with cytokeratin polypeptides no. 8 and 18 of Moll's catalogue; SK 2-27, specific for polypeptides no. 14, 16 and 17). The normal urothelial pattern is in agreement with previous reports. In pathological conditions, modified immunostaining has been demonstrated in almost all cases. In detail, the cytoskeletal pattern detected in transitional cell papilloma seems to discriminate between types which are otherwise histologically similar. We also observed a correlation between higher degrees of malignancy and loss of specialization, as demonstrated by the increasing positivity for SK 60-61, which as a rule specifically stains "umbrella" cells, and SK 2-27, an antibody exclusively detected in cells of the basal layer. These findings indicate that the cytokeratin pattern may constitute a modern new tool for the pathologist in the diagnosis of urothelial proliferative disorders.
Assuntos
Queratinas/metabolismo , Doenças da Bexiga Urinária/metabolismo , Bexiga Urinária/metabolismo , Anticorpos Monoclonais , Carcinoma de Células de Transição/metabolismo , Cistite/metabolismo , Epitélio/metabolismo , Histocitoquímica , Humanos , Imuno-Histoquímica , Metaplasia/metabolismo , Papiloma/metabolismo , Neoplasias da Bexiga Urinária/metabolismoRESUMO
Adenosine (10(-9)-10(-6) mol/l) and R-phenylisopropyladenosine (10(-9)-10(-7) mol/l) partially inhibited the intracellular accumulation of cyclic AMP induced by isoproterenol, prostaglandin E1, histamine and 5'-N-ethylcarboxamidoadenosine in lymphocytes. In contrast, S-phenylisopropyladenosine, which is a poor agonist of the adenosine A1/Ri receptor, had essentially no inhibitory effect. 8-Phenyltheophylline, in low concentrations that do not inhibit cyclic AMP phosphodiesterase, completely blocked the inhibitory effect of R-phenylisopropyladenosine on the increase in cyclic AMP induced by prostaglandin E1. R-Phenylisopropyladenosine (10(-8)-10(-6) mol/l) also inhibited the cyclic AMP accumulation in lymphocytes induced by forskolin (10(-5) mol/l), which activates adenylate cyclase through direct interaction with the enzyme. We also investigated the presence of the adenosine A1/Ri receptor on human polymorphonuclear leukocytes. R-Phenylisopropyladenosine (3 x 10(-9)-10(-7) mol/l) abolished the stimulating effects of prostaglandin and forskolin on cyclic AMP accumulation in polymorphonuclear leukocytes. This effect was blocked by 8-phenyltheophylline and was not observed with the stereoisomer S-phenylisopropyladenosine. The results support the existence of an A1/Ri receptor that regulates cyclic AMP metabolism of human lymphocytes and polymorphonuclear leukocytes.
Assuntos
Linfócitos/química , Neutrófilos/química , Receptores Purinérgicos/análise , Adenilil Ciclases/efeitos dos fármacos , Colforsina/antagonistas & inibidores , AMP Cíclico/análise , Ativação Enzimática/efeitos dos fármacos , Humanos , Subpopulações de Linfócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/enzimologia , Neutrófilos/enzimologia , Fenilisopropiladenosina/antagonistas & inibidores , Fenilisopropiladenosina/farmacologia , Teofilina/análogos & derivados , Teofilina/farmacologiaRESUMO
The large extracellular domain of CD81, a member of the tetraspanin family and a receptor protein for hepatitis C virus envelope E2 glycoprotein, has been expressed, purified and subsequently crystallized using the sitting-drop vapour-diffusion technique. Native diffraction data to 1.6 A resolution were obtained at the ID14 beamline of the European Synchrotron Radiation Facility from a flash-frozen crystal at 100 K. The crystals belong to space group P2(1), with unit-cell parameters a = 31.5, b = 77.2, c = 38.5 A, beta = 107.4 degrees, and are likely to contain two extracellular domains (2 x 99 residues) per asymmetric unit.
Assuntos
Antígenos CD/química , Hepacivirus/fisiologia , Proteínas de Membrana , Receptores Virais/química , Antígenos CD/fisiologia , Sequência de Bases , Cristalização , Cristalografia por Raios X , Primers do DNA , Humanos , Receptores Virais/fisiologia , Proteínas Recombinantes/química , Tetraspanina 28RESUMO
Recently, regional changes of cytokeratin patterns in human normal non-keratinized or keratinized oral mucosa have been demonstrated and the expression of individual cytokeratin polypeptides in lesions of oral mucosa has been compared with that of normal tissues. In particular, the presence of cytokeratin 19 in the suprabasal cell layers of oral epithelia has been shown to be strongly correlated with premalignancy. In the present study, we describe the results of an immunohistochemical investigation performed using a monoclonal antibody specific for cytokeratin 1 on normal oral mucosa and benign or malignant oral lesions. We show the different distribution of this polypeptide in non-neoplastic lesions from different sites of oral mucosa and describe the presence of cytokeratin 19. Our results are in agreement with the data obtained previously. In the malignant cases we demonstrate that the distribution of the two cytokeratins is characterized by complementary patterns.
Assuntos
Queratinas/metabolismo , Mucosa Bucal/metabolismo , Neoplasias Bucais/metabolismo , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Histocitoquímica , Humanos , Immunoblotting , Técnicas Imunoenzimáticas , Leucoplasia Oral/metabolismo , Leucoplasia Oral/patologia , Mucosa Bucal/patologia , Neoplasias Bucais/patologia , Papiloma/metabolismo , Papiloma/patologia , Lesões Pré-Cancerosas/metabolismo , Lesões Pré-Cancerosas/patologia , Distribuição TecidualRESUMO
A series of 23 punch biopsies proved to contain human papillomavirus (HPV) type 16 and with established clinical course (including HPV-NCIN, HPV-CIN I, and HPV-CIN II lesion), and 18 additional biopsies of HPV 6-, 11-, 16- or 18-induced genital lesions were analyzed immunohistochemically for expression of cytokeratin No. 19 polypeptide. An immunoperoxidase-ABC technique was used with a polyclonal antibody raised against a synthetic nonapeptide corresponding to the residues 2-10 of the NH2-end, non-alpha-helical region. This polyclonal cytokeratin No. 19 antibody stained mainly (but not exclusively) the basal cells of the normal exocervical epithelium (heterogeneous pattern). Basal cell staining was intense slightly more frequently in HPV-CIN than HPV-NCIN lesions, i.e., ++ or more in 14/24 (58.3%) versus 8/17 (47.0%), respectively. The difference was more marked in the staining of the superficial cells, 70.8 and 58.8% showing intense expression of cytokeratin No. 19, respectively. In 6 (21.4%) of the 28 HPV 16 lesions, basal cell layer was intensely stained, as contrasted to none of the 13 HPV 6, 11 or 18 lesions. The most distinct feature was the well-defined granular staining pattern of the superficial layer in 8 out of 10 HPV 6/11 lesions, as contrasted to the homogeneous pattern in 24 out of 28 HPV-16-infected lesions. In superficial cells, regressed lesions exhibited intense staining in 9/13 (69.2%), as compared with only 4/10 (40%) of the progressed lesions.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Doenças dos Genitais Femininos/metabolismo , Queratinas/análise , Infecções Tumorais por Vírus/metabolismo , Colo do Útero/metabolismo , Epitélio/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Papillomaviridae , Peptídeos/análise , Coloração e Rotulagem , Neoplasias do Colo do Útero/metabolismoRESUMO
Prolonged incubation (18h) of human polymorphonuclear leukocytes (PMNs) with betamethasone (BT) (10(-7) M) increased (3H)-DHA binding to human PMN membranes, apparently causing an increase in the number of beta-adrenergic receptors. Inhibition of this effect by cycloheximide (2 micrograms/ml) suggests that it is dependent on protein synthesis. BT had no effect on basal levels of cAMP in PMNs, but synergistically potentiated the increase in cyclic AMP (cAMP) induced by isoproterenol. Propranolol completely abolished the synergistic effect of BT plus isoproterenol, suggesting that the potentiating effect of BT on cAMP metabolism required the activation of beta-adrenergic receptors. Thus, BT increased the number of beta-adrenergic receptors in human PMN membranes and potentiated the cAMP changes induced by beta-agonists.