RESUMO
The Gynecologic Cancer InterGroup (GCIG) Fifth Ovarian Cancer Consensus Conference (OCCC) was held in Tokyo, Japan from 7 to 9 November 2015. It provided international consensus on 15 important questions in 4 topic areas, which were generated in accordance with the mission statement to establish 'International Consensus for Designing Better Clinical Trials'. The methodology for obtaining consensus was previously established and followed during the Fifth OCCC. All 29 clinical trial groups of GCIG participated in program development and deliberations. Draft consensus statements were discussed in topic groups as well as in a plenary forum. The final statements were then presented to all 29 member groups for voting and documentation of the level of consensus. Full consensus was obtained for 11 of the 15 statements with 28/29 groups agreeing to 3 statements, and 27/29 groups agreeing to 1 statement. The high acceptance rate of the statements among trial groups reflects the fact that we share common questions, and recognise important unmet needs that will guide future research in ovarian cancer.
Assuntos
Neoplasias Ovarianas/terapia , Feminino , Humanos , Avaliação das Necessidades , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Salmonella ovarian abscess in a patient with rheumatoid arthritis (RA) is reported here. A 33-year-old nulliparous woman with a 16-year history of RA who had been treated with corticosteroid and immunosuppressive drugs was diagnosed as having a non-typhoidal Salmonella ovarian abscess which might have been preceded by an occurrence of endometriotic cyst. Multidisciplinary therapy including surgical intervention was required to complete the eradication of infection. Although Salmonella ovarian abscess is rare, it may cause a serious complication in the ovary harboring endometriotic cyst through sustained presence of Salmonella bacteraemia.
Assuntos
Abscesso/microbiologia , Artrite Reumatoide/complicações , Doenças Ovarianas/microbiologia , Salmonella enteritidis , Adulto , Antibacterianos/administração & dosagem , Ceftriaxona/administração & dosagem , Feminino , Humanos , Lincomicina/administração & dosagem , Infecções por SalmonellaRESUMO
Choriocarcinoma is categorized as either gestational or nongestational depending on its origin. Nongestational choriocarcinoma originated in the trophoblastic differentiation is a rare but an aggressive tumor. This article reports a nongestational case of a uterine endometrial carcinoma with trophoblastic differentiation. A 54-year-old woman with a history of atypical genital bleeding that underwent semi-radical hysterectomy, bilateral salpingo-oophrectomy, and pelvic lymph nodes dissection. Pathological investigation showed that the tumor had endometrioid adenocarcinoma and choriocarcinomatous components. Although a series of multimodality treatments including craniotomy were performed, she died of aggressive lung and brain metastases one year after the primary surgery.
Assuntos
Carcinoma Endometrioide/patologia , Coriocarcinoma não Gestacional/patologia , Neoplasias do Endométrio/patologia , Tumor Misto Maligno/patologia , Carcinoma Endometrioide/diagnóstico por imagem , Carcinoma Endometrioide/cirurgia , Coriocarcinoma não Gestacional/diagnóstico por imagem , Coriocarcinoma não Gestacional/cirurgia , Neoplasias do Endométrio/diagnóstico por imagem , Neoplasias do Endométrio/cirurgia , Feminino , Humanos , Histerectomia , Pessoa de Meia-Idade , Tumor Misto Maligno/diagnóstico por imagem , Tumor Misto Maligno/cirurgia , Ovariectomia , Salpingectomia , Tomografia Computadorizada por Raios XRESUMO
BACKGROUND: Recently, the vitamin D receptor (VDR) polymorphism FokI was shown to be associated with susceptibility to ovarian cancer. We aimed to examine whether VDR FokI polymorphisms influence the survivals of patients with epithelial ovarian cancer (EOC). METHODS: VDR polymorphisms from FokI in 101 patients with EOC were genotyped by sequencing. Overall survival was compared between FokI single nucleotide polymorphism using Kaplan-Meier survival curves with log-rank tests and the Cox proportional hazard model adjusted for ages, stages, histology, and existence of residual tumour. RESULTS: The FokI C/C genotypes were associated with better prognosis compared with the C/T and T/T genotypes (log-rank test: P = 0.008; adjusted hazard ratio, 0.18; 95%CI 0.05-0.61; P = 0.006). CONCLUSIONS: These results suggest that the VDR polymorphisms from the FokI genotype may be associated with improved prognosis of patients with EOC.
Assuntos
Neoplasias Epiteliais e Glandulares/genética , Neoplasias Ovarianas/genética , Polimorfismo de Nucleotídeo Único , Receptores de Calcitriol/genética , Adulto , Estudos de Coortes , Desoxirribonucleases de Sítio Específico do Tipo II/metabolismo , Feminino , Genótipo , Humanos , Pessoa de Meia-Idade , Neoplasias Epiteliais e Glandulares/mortalidade , Neoplasias Ovarianas/mortalidade , Prognóstico , Modelos de Riscos Proporcionais , Vitamina D/análogos & derivados , Vitamina D/sangueRESUMO
Chromogranin A (CgA) is an acidic glycoprotein that is co-stored with hormones or neurotransmitters in granular components of endocrine cells and neurons, and released together with them in response to adequate stimulation. In addition to acting as a packaging protein, CgA functions as a precursor molecule that yields several bioactive peptides by proteolytic cleavage. The purpose of this study is to elucidate how different the processing of CgA is among endocrine tissues by immunostaining using multiple region-specific antisera, and to evaluate the availability of region-specific antisera. When various endocrine organs of rats were immunostained with four region-specific antisera against rat CgA (CgA 1-28, 94-130, 296-314, and 359-389), all amine/peptide-secreting endocrine tissues except the pineal body were stained positively. The adrenal medulla and gastric endocrine cells were equally intensely immunoreactive to all four antisera, while the other endocrine tissues, represented by pancreatic islets, showed different staining patterns depending on the antiserum. These results suggest that the processing of CgA differs from tissue to tissue. An antiserum against horse CgA 335-365, corresponding to rat CgA 359-389 which shows the highest concentration in the plasma and urine of the rat, again stained all endocrine tissues of the horse except the pineal body. Therefore, the anti-horse CgA 335-365 serum is useful for immunohistochemical survey of horse CgA, and may make possible the establishment of a CgA assay system for the measurement of CgA in the plasma, urine and saliva.
Assuntos
Cromograninas/análise , Glândulas Endócrinas/química , Animais , Cromogranina A , Feminino , Cavalos , Soros Imunes , Imuno-Histoquímica/veterinária , Masculino , Radioimunoensaio/veterinária , Ratos , Ratos WistarRESUMO
This immunohistochemical study analyzed the c-Fos expression (c-Fos-ir) induced by galanin injections. Galanin and N-terminal galanin fragment (1-15) induced a significant increase of c-Fos expression (c-ir) within the medulla oblongata 90 min and 6 h. after intracisternal injections. This expression has been studied mainly in the nucleus of the solitary tract and in the ventrolateral medulla showing different temporal profiles for both peptides. The presence of c-Fos-ir in TH-positive cells was analyzed in all the groups. These results may be relevant to understand the role of galanin in several functions including central cardiovascular control.
Assuntos
Galanina/farmacologia , Bulbo/metabolismo , Fragmentos de Peptídeos/farmacologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Núcleo Solitário/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Contagem de Células , Proteína Glial Fibrilar Ácida/imunologia , Imuno-Histoquímica , Masculino , Bulbo/efeitos dos fármacos , Bulbo/enzimologia , Neuroglia/imunologia , Neurônios/imunologia , Neurônios/metabolismo , Proteínas Proto-Oncogênicas c-fos/imunologia , Coelhos , Ratos , Ratos Sprague-Dawley , Núcleo Solitário/citologia , Núcleo Solitário/efeitos dos fármacos , Núcleo Solitário/enzimologia , Organismos Livres de Patógenos Específicos , Fatores de Tempo , Tirosina 3-Mono-Oxigenase/imunologiaRESUMO
Here, we report the complete genomic sequence and the characterization of the 311-kb region of 18q21, a candidate tumor suppressor locus containing a region of homozygous deletion in a lung cancer cell line, Ma29. This region contained two known genes, SMAD4 and ME2 (mitochondrial malate oxydoreductase), and two novel genes, D29 (deleted in Ma29 HGMW-approved symbol ELAC1), encoding an evolutionarily conserved protein, and B29 (beside the Ma29 deletion HGMW-approved symbol C18orf3), with no significant homology to any known genes. The deleted DNA segment in Ma29, which was estimated to be 195 kb in size, included all the coding exons of ME2 and D29, but not the coding exons of SMAD4 and B29. The deleted region also included exon 0, a 5'-noncoding exon, of SMAD4, and the expression of SMAD4 was greatly reduced in Ma29 cells. Mutations of SMAD4 and D29 were detected in 1 of 45 lung cancer cell lines examined, while those of ME2 and B29 were not detected, indicating that these four genes are not major targets for 18q21 deletions. The physical and transcriptional map constructed in this study will provide basic information for the identification of a tumor suppressor gene(s) at 18q21 involved in lung carcinogenesis.
Assuntos
Cromossomos Humanos Par 18 , Genes Supressores de Tumor , Neoplasias Pulmonares/genética , Proteínas Supressoras de Tumor , Antígenos CD/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Antígenos CD79 , Mapeamento de Sequências Contíguas , DNA de Neoplasias , Proteínas de Ligação a DNA/genética , Deleção de Genes , Expressão Gênica , Humanos , Dados de Sequência Molecular , Mutação , Proteínas de Neoplasias/genética , Proteínas do Tecido Nervoso/genética , Análise de Sequência de DNA , Proteína Smad4 , Fatores de Transcrição TCF , Transativadores/genética , Fator de Transcrição 4 , Fatores de Transcrição/genética , Transcrição Gênica , Células Tumorais CultivadasRESUMO
Chromogranin A (CGA) is a member of a family of highly acidic proteins co-stored and co-released with catecholamines in the adrenal medullary cells as well as in other neurons and paraneurons. The nucleotide sequence encoding equine CGA was determined using RT-PCR and rapid amplification of complementary DNA (cDNA) ends (RACE) techniques. A total 1,828 bp of the nucleotide sequence reveals that equine CGA is a 448-residue protein preceded by an 18-residue signal peptide. Comparison of the amino acid sequence of equine CGA with those of human, porcine, bovine, mouse, rat and frog CGA showed high conservation at the NH2-terminal 1-77 amino acids regions (94.8%, 93.5%, 92.2%, 81.8%, 83.1% and 66.2%, respectively) and COOH-terminal 314-430 amino acids regions (90.6%, 81.4%, 90.6%, 80.5%, 83.3% and 39.0%, respectively), as well as a potential dibasic cleavage site, whereas the middle portion showed marked sequence variation (52.5%, 49.1%, 38.9%, 26.6%, 27.9% and 6.2%, respectively). Northern blot analysis and RT-PCR elucidated the tissue distribution of equine CGA mRNA. Its expression was confirmed not only in the adrenal medullary cells but also in other organs (cerebrum, cerebellum, pituitary gland, spinal cord, liver, thyroid gland, striated muscle, lung, spleen, kidney, parotid gland and sublingual gland). Further, in adrenal chromaffin cells and pituitary cells of the anterior-intermediate lobe, the expression was confirmed by in situ hybridization with anti-sense CGA cRNA probe.
Assuntos
Cromograninas/genética , Regulação da Expressão Gênica , Sequência de Aminoácidos , Animais , Anuros , Sequência de Bases , Northern Blotting/veterinária , Bovinos , Cromogranina A , Cromograninas/biossíntese , Clonagem Molecular , DNA Complementar/química , Glândulas Endócrinas/metabolismo , Glândulas Exócrinas/metabolismo , Cavalos , Humanos , Hibridização In Situ/veterinária , Camundongos , Dados de Sequência Molecular , RNA Mensageiro/química , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , SuínosRESUMO
Glicentin (GLIC), oxyntomodulin (OXM), and peptide YY (PYY) released in blood by ileocolonic L-cells after meals may inhibit pancreatic secretion. Whereas OXM interacts with glucagon and tGLP-1 receptors, OXM 19-37, a biologically active fragment, does not. The purpose of this study was to measure the effect of OXM, OXM 19-37, GLIC, tGLP-1, and PYY on pancreatic secretion stimulated by 2 deoxyglucose (2DG), electrical stimulation of the vagus nerves (VES), acetylcholine and cholecystokinin octapeptide (CCK8) in anesthetized rats. The effect of OXM was also studied in dispersed pancreatic acini. Plasma oxyntomodulin-like immunoreactivity (OLI) was measured by radioimmunoassay after the exogenous infusion of OXM and after an intraduodenal meal. OXM 19-37, infused at doses mimicking postprandial plasma levels of OLI, decreased pancreatic secretion stimulated by 2DG, VES, or CCK8. Similar effects were found with OXM and GLIC. OXM 19-37 did not change the pancreatic stimulation induced by acetylcholine in vivo, or CCK-induced amylase release in isolated acini. Vagotomy completely suppressed the inhibitory effect of OXM 19-37 on CCK8-stimulated pancreatic secretion. PYY inhibited the effect of 2DG, but not that of CCK8, whereas tGLP-1, even in pharmacologic doses, had no effect on stimulated pancreatic secretion. OXM, OXM 19-37, but not tGLP-1, inhibit pancreatic secretion at physiologic doses, through a vagal neural indirect mechanism, different from that used by PYY, and probably through a GLIC-related peptide-specific receptor.
Assuntos
Peptídeos Semelhantes ao Glucagon/farmacologia , Pâncreas/inervação , Pâncreas/metabolismo , Animais , Desoxiglucose/farmacologia , Interações Medicamentosas , Glicentina , Glucagon/farmacologia , Peptídeo 1 Semelhante ao Glucagon , Peptídeos Semelhantes ao Glucagon/sangue , Cinética , Masculino , Oxintomodulina , Pâncreas/efeitos dos fármacos , Fragmentos de Peptídeos/sangue , Fragmentos de Peptídeos/farmacologia , Peptídeo YY/sangue , Peptídeo YY/farmacologia , Precursores de Proteínas/farmacologia , Ratos , Ratos Wistar , Sincalida/farmacologia , Nervo Vago/efeitos dos fármacos , Nervo Vago/fisiologiaRESUMO
The modulation of the central cardiovascular effects of alpha(2)-adrenoceptor activation by galanin and its N-terminal fragment galanin-(1-15) has been evaluated by quantitative receptor autoradiography and cardiovascular analysis. Intracisternal coinjections of threshold doses of galanin and the selective and hypotensive alpha(2)-receptor agonist clonidine induced rapid and maintained vasopressor and tachycardic responses (p<0.001) instead of a hypotensive response, whereas the coinjections of threshold doses of the N-terminal galanin fragment (1-15) and clonidine did not elicit significant cardiovascular changes. Receptor autoradiographical experiments showed that galanin (1 nM) significantly increased the K(d) (p<0.01) and B(max) values (p<0.01) of [(3)H]p-Aminoclonidine binding sites in the nucleus tractus solitarii (NTS) compatible with a possible antagonistic interaction with the alpha(2)-adrenoceptors, and this effect was blocked by the presence of the specific galanin receptor antagonist M35. In addition, clonidine (30 nM) induced a 50% increase in the B(0) values of galanin based on competition experiments with [(125)I]-galanin binding in the NTS. These findings suggest the existence of an antagonistic effect of galanin, but not of galanin fragment (1-15), on the cardiovascular responses mediated by alpha(2)-receptors as well as a reciprocal facilitatory effect of alpha(2)-receptors on galanin binding. These mechanisms could be mediated by a reciprocal galanin-alpha(2) receptor interaction within the NTS.
Assuntos
Fenômenos Fisiológicos Cardiovasculares/efeitos dos fármacos , Galanina/farmacologia , Receptores Adrenérgicos alfa 2/metabolismo , Agonistas alfa-Adrenérgicos/farmacologia , Animais , Autorradiografia , Ligação Competitiva/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Bradicinina/análogos & derivados , Bradicinina/farmacologia , Clonidina/análogos & derivados , Clonidina/metabolismo , Clonidina/farmacologia , Relação Dose-Resposta a Droga , Galanina/metabolismo , Cinética , Masculino , Fragmentos de Peptídeos/farmacologia , Ligação Proteica , Ratos , Ratos Sprague-Dawley , Receptores de Galanina , Receptores de Neuropeptídeos/antagonistas & inibidores , Núcleo Solitário/efeitos dos fármacos , Núcleo Solitário/metabolismo , Organismos Livres de Patógenos Específicos , TrítioAssuntos
Cromograninas/genética , DNA Complementar/genética , Cavalos/genética , Cavalos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Animais , Cromogranina A , Cromograninas/metabolismo , Glândulas Endócrinas/metabolismo , Glândulas Exócrinas/metabolismo , Imuno-Histoquímica , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Glândula Submandibular/metabolismo , Distribuição TecidualAssuntos
Sistema Nervoso Autônomo/fisiologia , Cromograninas/metabolismo , Glândulas Salivares/metabolismo , Acetilcolina/farmacologia , Animais , Sistema Nervoso Autônomo/efeitos dos fármacos , Cromogranina A , Humanos , Imunoensaio , Imuno-Histoquímica , Camundongos , Norepinefrina/farmacologia , Peptídeos/metabolismo , Ratos , Glândulas Salivares/efeitos dos fármacos , Estresse Fisiológico/fisiopatologiaRESUMO
In the study reported in this paper, sensitive ELISA for rat CgA was developed using synthetic rat CgA(359-389) as antigen, N alpha-biotinylated glycylglycyl rat CgA(359-389), and antirat CgA(359-389) serum for the measurement of CgA-LI in rat saliva. CgA-LI in rat submandibular tissues and saliva was characterized by both immunohistochemical and immunochemical methods. Using isolated perfused rat submandibular gland. VIP at 0.1-1.0 nM in the presence of 0.1 microM ACh was found to cause CgA-LI secretion, whereas neither PACAP-27 nor PACAP-38 showed any effect on CgA secretion.
Assuntos
Cromograninas/metabolismo , Neuropeptídeos/farmacologia , Glândula Submandibular/efeitos dos fármacos , Glândula Submandibular/metabolismo , Peptídeo Intestinal Vasoativo/farmacologia , Sequência de Aminoácidos , Animais , Cromogranina A , Cromograninas/química , Ensaio de Imunoadsorção Enzimática/métodos , Imuno-Histoquímica , Técnicas In Vitro , Dados de Sequência Molecular , Neuropeptídeos/metabolismo , Fragmentos de Peptídeos/química , Perfusão , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase , Ratos , Saliva/metabolismo , Peptídeo Intestinal Vasoativo/metabolismoAssuntos
Glucagon/farmacologia , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/fisiologia , Fragmentos de Peptídeos/farmacologia , Peptídeos/farmacologia , Precursores de Proteínas/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Animais , DNA/metabolismo , Glicentina , Peptídeo 1 Semelhante ao Glucagon , Peptídeo 2 Semelhante ao Glucagon , Peptídeos Semelhantes ao Glucagon , Substâncias de Crescimento/farmacologia , Intestino Delgado/cirurgia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Proteínas/metabolismo , Ratos , Ratos Wistar , Sacarase/metabolismo , Fatores de Tempo , alfa-Glucosidases/metabolismoRESUMO
Chromogranin A (CgA) is a member of a family of highly acidic proteins, chromogranins, which are co-stored in the adrenergic neurons and paraneurons and co-released with adrenaline and noradrenaline (NAd) in response to adequate stimulation. The present study provides novel evidence that CgA-like immunoreactivity (IR) is stored in the exocrine cells in the granular convoluted tubule, and is secreted into saliva by stimulation with NAd and acetylcholine (ACh) in the isolated and perfused rat submandibular gland. NAd at 1 microM produced maximum secretion of CgA-like IR (<< 0.9 mM) and a marked increase in salivary flow. Further increases in NAd concentration (10 or 100 microM) yielded concentration-dependent decreases in both responses. ACh at 1 microM produced maximum salivary flow and a slight elevation of CgA-like IR secretion (6 microM); 100 microM ACh decreased the salivary flow but increased the CgA-like IR secretion (0.6 mM). Electron microscopic examination showed vigorous compound exocytosis of secretory granules in the cells of the granular convoluted tubule when the submandibular gland was stimulated with 1 microM NAd. These results provide an experimental basis for the view that the salivary CgA-like IR secretion may be a sensitive and quantitative index of the activity of the sympathetic nervous system innervating the gland.
Assuntos
Acetilcolina/farmacologia , Cromograninas/análise , Norepinefrina/farmacologia , Saliva/metabolismo , Glândula Submandibular/efeitos dos fármacos , Animais , Cromogranina A , Masculino , Microscopia Eletrônica , Perfusão , Radioimunoensaio , Ratos , Ratos Wistar , Saliva/química , Estresse Psicológico , Glândula Submandibular/metabolismo , Glândula Submandibular/ultraestruturaRESUMO
P-glycoprotein (Pgp) is a plasma-membrane glycoprotein that confers multi-drug resistance (MDR) on cells and displays ATP-driven drug pumping. The possible contribution of calpain-mediated proteolytic pathways to the functional regulation of the Pgp molecule was evaluated using K562/DXR, MDR cells. N-Acetyl-L-leucyl-L-leucyl-norleucinal was effluxed by Pgp, but N-benzyloxycarbonyl-L-leucyl-L-leucinal (zLLal), an inhibitor of calpain, retarded the degradation of Pgp leading to accumulation of the molecule largely at the cell surface membrane. Treatment with brefeldin A did not obstruct the zLLal-induced Pgp accumulation. NH4Cl increased the cytoplasmic Pgp level, with a slight to significant decrease at the cell surface membrane. Ubiquitin-ELISA and western blot analysis confirmed that the Pgp molecule, which accumulated mainly at the cell surface, was ubiquitinated. However, lactacystin did not show any accumulation of Pgp in either the cytoplasm or the cell surface membrane, suggesting that the proteasome did not participate in the phenomenon. Additionally, the Pgp was limitedly proteolyzed by calpain into two 98 kDa and 69 kDa, fragments within one minute. Despite the increased accumulation of Pgp at the cell surface after treatment with calpain inhibitor, the cytoplasmic doxorubicin level of the cells treated with a calpain inhibitor was higher than that of non-treated cells and approached that of parental cells. These results indicated that calpain involved Pgp turnover and that calpain inhibition induced ubiquitinated Pgp-accumulation mainly at the cell surface membrane with a reduction in its own functions suggesting that the modulation of Pgp-turnover involves MDR-reversal by another approach.
Assuntos
Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Calpaína/antagonistas & inibidores , Calpaína/fisiologia , Inibidores de Cisteína Proteinase/farmacologia , Glicoproteínas/farmacologia , Acetilcisteína/análogos & derivados , Acetilcisteína/farmacologia , Cloreto de Amônio/farmacologia , Western Blotting , Brefeldina A/farmacologia , Calpaína/metabolismo , Membrana Celular/metabolismo , Dipeptídeos/farmacologia , Doxorrubicina/metabolismo , Ensaio de Imunoadsorção Enzimática , Humanos , Células K562 , Leucina/metabolismo , Leupeptinas/farmacologia , Proteínas de Membrana/metabolismo , Inibidores da Síntese de Proteínas/farmacologia , Fatores de Tempo , Ubiquitinas/metabolismoRESUMO
Chromogranin A (CgA), a secretory protein, is co-released with catecholamines from storage vesicles. It is known to be elevated in the circulation of patients with neuroendocrine and endocrine tumors. For further investigation of the protein, especially in humans, it is essential to facilitate quantitative analysis of the protein in human biological materials. In order to introduce novel immunological methodology for this purpose, we purposely selected human CgA(344-374) for the synthetic immunogen to produce region-specific CgA antibodies. The anti-synthetic peptide antibody thus obtained made it possible to develop an immunological method for measurement and characterization of CgA in human plasma. The plasma CgA-immunoreactivity (LI) level measured by the method was 0.31+/-0.01 pmol/ml (mean+/-SEM) in normal subjects and 1.55+/-0.29 pmol/ml in pheochromocytoma. On gel chromatography and HPLC analysis of the plasma of patients with pheochromocytoma, the region-specific assay system enabled us to show the presence of N-terminal truncated CgA, besides CgA itself. By following up changes of plasma CgA-LI in a pheochromocytoma patient using samples that were collected consecutively over a two-year period, the present assay system using the region-specific antibody, anti-human CgA (344-374) serum, was confirmed to be extremely valuable for the measurement of CgA-LI in human plasma. The characteristic features and high sensitivity of the present assay system will give us a substantial clue to the detection and measurement of CgA to develop further investigation of the protein in humans.
Assuntos
Cromograninas/sangue , Cromograninas/imunologia , Imunoensaio/métodos , Neoplasias das Glândulas Suprarrenais/metabolismo , Neoplasias das Glândulas Suprarrenais/terapia , Adulto , Idoso , Sequência de Aminoácidos , Formação de Anticorpos , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia Líquida/métodos , Cromogranina A , Dopamina/sangue , Epinefrina/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Norepinefrina/sangue , Feocromocitoma/metabolismo , Feocromocitoma/terapia , Valor Preditivo dos Testes , Valores de Referência , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Complete amino acid sequences of two novel bioactive polypeptides, each containing 66 amino acid residues, BmK AS and BmK AS-1 purified from the venom of Chinese scorpion Buthus martensi Karsch, have been determined by Edman sequencing and mass spectrometry on native proteins, reduced and S-carboxymethylated proteins and their peptides obtained after cleavage with proteolytic enzymes. Sequence analysis showed 86.4% structural identity between BmK AS and BmK AS-1 and also a high sequence similarity between BmK ASs and AaH IT4, a unique anti-insect toxin and a ligand of Na+ channels obtained from Sahara scorpion A. australis Hector, but poor sequence homology between BmK ASs and those of the known alpha-, beta-type and long-chain insect-selective type scorpion neurotoxins. The positions of four disulfide bridges in BmK AS-1 were established as Cys-12 and Cys-62, Cys-16 and Cys-37, Cys-23 and Cys-44, and Cys-27 and Cys-46, which are the same as those in alpha- and beta-scorpion neurotoxins. These results suggest that BmK ASs and AaH IT4 may form a new group sharing similar structural and functional properties in the family of scorpion neurotoxic polypeptides.
Assuntos
Neurotoxinas/química , Peptídeos/química , Venenos de Escorpião/química , Homologia de Sequência de Aminoácidos , Sequência de Aminoácidos , Animais , Ligantes , Espectrometria de Massas , Dados de Sequência Molecular , Peptídeo Hidrolases , Peptídeos/isolamento & purificação , Venenos de Escorpião/isolamento & purificação , Escorpiões , Canais de SódioRESUMO
A luminal cholecystokinin releasing factor (LCRF), has been purified from intestinal secretion and found to have a mass of 8136 daltons. The amino-terminal 41 residues have been sequenced. Previous studies showed that intraduodenal infusion of the synthetic amino-terminal 35 amino acid peptide, LCRF1-35 significantly stimulated pancreatic protein and fluid secretion in conscious rats, but the peptide did not stimulate amylase release from isolated, dispersed pancreatic acini. In the present study, several fragments of LCRF were synthesized and tested for CCK-releasing activity (pancreatic protein secretion) to determine whether shorter fragments of LCRF exhibit the characteristic biological activity of native LCRF and synthetic LCRF1-35. Compounds tested were LCRF1-41, LCRF1-35, LCRF1-65 and LCRF11-25. Of the fragments shorter than LCRF1-35, only LCRF11-25 but not LCRF1-6 had significant CCK releasing activity. LCRF1-41 was equivalent to LCRF1-35 in potency and efficacy. Intravenous and intraduodenal infusion of LCRF1-35 elicited nearly identical dose-response curves.