RESUMO
The ovarian steroids estrogen and progesterone are important in directing the normal growth and development of the mouse mammary gland. Previously, we have demonstrated that the majority of proliferating mammary epithelial cells do not express estrogen receptor-alpha (ERalpha). In this study we examined the relationship between progesterone receptor (PR) expression and proliferation in mammary epithelial cells using simultaneous immunohistochemistry for progesterone receptor (PR) and tritiated thymidine [(3)H]-Tdr) autoradiography. Results showed that the majority (>80%) of mammary epithelial cells labeled with [(3)H]-Tdr were PR-positive in the terminal end buds (TEBs) of pubertal mice and the ducts of pubertal and adult mice. Whereas the majority of mammary epithelial cells were also PR-positive, the basal cell population, which comprises the minority of mammary epithelial cells in the mammary ducts, was predominantly PR-negative. Nevertheless, the PR-positive phenotype remained the major proliferating cell type in the basal population. These findings suggest that the progesterone signaling pathway is involved in the proliferation of basal cell populations, potentially directing formation of tertiary side branching during pubertal development and alveolar bud formation in adult glands. A proportion of the basal cells exhibited weak expression of ERbeta, suggesting that the role of ERbeta in mediating normal estrogen-induced responses should be further studied. (J Histochem Cytochem: 47:1323-1330, 1999)
Assuntos
Estro/metabolismo , Glândulas Mamárias Animais/metabolismo , Receptores de Estrogênio/biossíntese , Receptores de Progesterona/biossíntese , Maturidade Sexual , Animais , Células Epiteliais/metabolismo , Receptor alfa de Estrogênio , Receptor beta de Estrogênio , Feminino , Técnicas Imunoenzimáticas , Glândulas Mamárias Animais/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Maintenance of the size and differentiated function of the adult prostate is dependent on testicular androgens. In this study, simultaneous androgen receptor (AR) immunohistochemistry and [(3)H]thymidine labelling was used to characterise the proliferating epithelial cells of the murine ventral prostate. Proliferation in the adult prostate was more prevalent in the basal cell population with 1.8&percent; AR-negative cells labelled with [(3)H]thymidine as compared with 0.7% AR-expressing luminal cells. Three weeks following castration of mice, the atrophied prostate contained rudimentary glands composed of both luminal and basal cells with the proportion of AR-expressing basal cells reduced from 50 to 25%. Administration of testosterone enanthate to castrated mice induced a recapitulation of the prostate gland that was preceded by up-regulation of AR expression in basal cells to normal adult levels (50% AR-positive cells) by 12 h following testosterone injection. Proliferation of AR-positive luminal cells peaked at 48 h (22.8%) while proliferation of AR-negative basal cells peaked at 96 h (6.1%) following testosterone administration. These results suggest that distinct populations of luminal and basal cells are resistant to castration-induced involution of the prostate but remain responsive to direct or indirect testosterone effects and recapitulate the gland following administration of testosterone.
Assuntos
Próstata/metabolismo , Receptores Androgênicos/metabolismo , Animais , Western Blotting , Contagem de Células , Divisão Celular , Células Cultivadas , Células Epiteliais/química , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Orquiectomia , Próstata/química , Próstata/efeitos dos fármacos , Receptores Androgênicos/análise , Receptores Androgênicos/genética , Testosterona/sangue , Testosterona/farmacologia , Fatores de TempoRESUMO
A breeding programme, based on selection for faecal egg counts, has produced lines of sheep which demonstrate either increased resistance (high responder) or susceptibility (low responder) to challenge infection with T. colubriformis after vaccination with irradiated larvae. Circulating blood eosinophilia, a hallmark of helminth infections, was examined in third generation lambs from two separate selective matings and random bred control lambs. Numbers of eosinophils were higher in high responder lambs when compared to low responders after vaccination and challenge infections. Analysis of eosinophil counts confirmed a strong line effect and there was no evidence of a sex effect. Random bred lambs showed wide individual variations in eosinophil numbers and their response to infection. It was concluded that peripheral eosinophilia was more a measure of host responsiveness to infection than an indicator of helminthiasis. As such the eosinophil may serve as an indicator of the hosts ability to respond to T. colubriformis vaccination and infection.
Assuntos
Eosinofilia/veterinária , Doenças dos Ovinos/imunologia , Tricostrongiloidíase/veterinária , Tricostrongilose/veterinária , Animais , Suscetibilidade a Doenças , Eosinofilia/imunologia , Feminino , Imunidade Inata , Contagem de Leucócitos/veterinária , Masculino , Fatores Sexuais , Ovinos , Tricostrongilose/imunologiaRESUMO
Cytoplasmic clarity is a histological feature of normal prostatic secretory cells, but in this study, tissue fixation in strong (>2.5%) glutaraldehyde dramatically altered cytological staining. Secretory cytoplasm appeared red and granular on routine stains because of myriad intensely staining eosinophilic granules (PSG). Immunostaining for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) showed their exclusive localization to the PSG. Electron microscopy confirmed these findings and also showed that after fixation in many agents, including formaldehyde, PSG appeared empty, accounting for the artefactual "clear cell" appearance on light microscopy. PSG were most densely concentrated apically in a bud-shaped luminal compartment in which cytokeratin was selectively absent. Normal exocrine secretion was visualized as detachment of apocrine buds or their in situ disintegration. Distinctively in dysplasia and almost all carcinomas, PSG were rare to absent, and proteases were free in the cytoplasm, often concentrated beneath the apical membrane. The apocrine compartment was absent, with no observed secretory mechanism. Tumor cells had dark amphiphilic cytoplasm after all fixatives. This provided a reliable method of distinguishing malignant from benign glands in tissues fixed in strong glutaraldehyde. Clear cell carcinomas, whose cytoplasm mimicked routinely fixed normal secretory cells, surprisingly had almost no PSG. Instead, their "granules" were lipid-filled vacuoles reflecting a secretory pathway not seen in normal cells, dysplasia, or the common "dark cell" carcinomas. These observations may define two distinctive biological pathways of prostate cancer evolution and may facilitate diagnostic decisions on needle biopsy samples.
Assuntos
Adenocarcinoma de Células Claras/patologia , Transformação Celular Neoplásica/patologia , Grânulos Citoplasmáticos/ultraestrutura , Lesões Pré-Cancerosas/patologia , Neoplasias da Próstata/patologia , Fosfatase Ácida/metabolismo , Adenocarcinoma de Células Claras/metabolismo , Grânulos Citoplasmáticos/metabolismo , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Humanos , Técnicas Imunoenzimáticas , Masculino , Lesões Pré-Cancerosas/metabolismo , Próstata/metabolismo , Próstata/patologia , Antígeno Prostático Específico/metabolismo , Neoplasias da Próstata/metabolismoRESUMO
We describe a sensitive and practical in situ hybridization method, using a digoxigenin-labeled probe, for the detection of c-erbB-2 amplification in breast cancer in formalin-fixed, paraffin-embedded tissue sections. Forty-six primary breast carcinomas were studied. Nuclear hybridization signal was observed in 36 of 46 carcinomas. Signal was confined to malignant cells. Normal breast epithelium and stromal and inflammatory cells were uniformally negative. DNase predigestion, no-probe preparations, and competitive hybridization confirmed the specificity of the reaction. The hybridization reaction was localized to multiple discrete foci in tumor cell nuclei, suggesting multiple sites of gene copy and transcriptional activity in the nucleus. Considerable cell-to-cell variation in hybridization signal was evident within individual tumors and positive reactions were observed in several cases in which amplification could not be detected by either Southern or slot blot analysis. The high sensitivity and specificity of the reaction and its use in a tissue-based system will allow the study of a range of possible precursor lesions of breast cancer for evidence of c-erbB-2 amplification.
Assuntos
Neoplasias da Mama/genética , Receptores ErbB/genética , Proteínas Proto-Oncogênicas/genética , Neoplasias da Mama/patologia , Amplificação de Genes , Humanos , Hibridização In Situ , Inclusão em Parafina , Receptor ErbB-2 , Fixação de TecidosRESUMO
Recent changes in tissue fixation strategy, using glutaraldehyde, have clarified the secretory mechanisms of the normal prostate identifying cytoplasmic prostatic secretory granules, structures not preserved by formalin fixation. This normal secretory mechanism was absent in most adenocarcinomas, depicting an important metabolic change in transformed prostate cells. The current study further investigates differences between benign and malignant prostate secretion and relates them to the production of corpora amylacea by benign glands and crystalloids or mucin by cancer. In all normal prostate cells examined (6 cases), prostate secretory granules (PSG) were approximately 1-microm, brightly eosinophilic granules filling the cytoplasm of secretory cells and released in packets by a specialized apocrine cell structure. After apocrine decapitation and luminal dispersal, some of the cytoplasmic and PSG remnants condensed to form eosinophilic bodies (EB) with a glycoprotein rim and central protein core. EB were observed adsorbing and layering onto the surface of prostatic corpora amylacea representing their chief mode of enlargement. Biochemical analysis and x-ray diffraction studies confirmed sulfated glycosaminoglycans of similar structure as the main constituent of both PSG and corpora amylacea. Peripheral zone amphiphilic "dark cell" carcinoma (9 cases) contained almost no PSG, and showed neither apical decapitation nor EB formation, but mucin secretion was frequently detected. Crystalloids that share the same staining characteristics and sulfur content as PSG and corpora amylacea were identified in 3 selected "clear cell" carcinomas, all of which showed at least focal PSG secretion. The recognition of these differing secretory mechanisms and their deviation from normal further defines the histological criteria and spectrum of prostate malignancy.
Assuntos
Carcinoma/metabolismo , Carcinoma/patologia , Próstata/metabolismo , Próstata/patologia , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Fosfatase Ácida/metabolismo , Citoplasma/ultraestrutura , Grânulos Citoplasmáticos/ultraestrutura , Humanos , Corpos de Inclusão/ultraestrutura , Masculino , Antígeno Prostático Específico/metabolismoRESUMO
Human prostatic secretion and seminal plasma contain certain protein kinase activities. Protein kinases play important roles in regulating a vast variety of cellular functions. The objective of this study was to determine whether one of these protein kinase activities in human prostatic secretion and seminal plasma is due to CK2, a messenger-independent, serine/threonine protein kinase that has considerable potential as a regulatory enzyme. By employing an anti-CK2 antibody and a CK2-specific peptide substrate, we have established that CK2 is present in these secretions. Approximately 70% of the CK2 activity present in seminal plasma of normozoospermic men (n = 49) is correlated to the number of sperm originally present in the semen. Further, both the prostate gland and the seminal vesicles are sources of CK2 activity in the seminal plasma of vasectomized men (n = 38). Although there was considerable variation between individuals in CK2 activity, the variation in repeat semen samples of the same vasectomized men (n = 6) was within 21%. There was no correlation of CK2 activity in seminal plasma with age for vasectomized (27-48 years, n = 38), oligozoospermic (28-43 years, n = 24), or normozoospermic men (26-48 years, n = 49). These data suggest that the majority of CK2 activity in the seminal plasma of normozoospermic men originates from sperm but that the prostate and seminal vesicles are accessory sex-gland sources of this enzyme.
Assuntos
Próstata/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Sêmen/enzimologia , Glândulas Seminais/enzimologia , Adulto , Sequência de Aminoácidos , Caseína Quinase II , Humanos , Masculino , Pessoa de Meia-Idade , FosforilaçãoRESUMO
Metastasis by thin melanomas is uncommon and unpredictable. In order to assess the prognostic value of p53 expression, p53 immunohistochemical staining was evaluated in 20 thin melanomas with documented metastasis and in 20 control tumours which failed to metastasize. Tumours selected were less than 1 mm thick and were individually matched for tumour thickness, date of excision and patient age and sex. The relative risk of metastasis given p53 overexpression was 1.5 (95% confidence interval 0.4-5.3; p = 0.53). Further quantitative analysis for p53 between the two groups did not demonstrate a significant difference (p = 0.08). These results are consistent with there being no association between p53 overexpression in thin melanomas and risk of metastasis, however, the sample size was small, and the existence of such an association cannot be ruled out with confidence. For the 20 thin melanomas which metastasized, there was no association between the proportion of cells positive for p53 and length of the relapse-free period (correlation coefficient = 0.02, p = 0.94).
Assuntos
Regulação Neoplásica da Expressão Gênica , Genes p53 , Melanoma/genética , Proteínas de Neoplasias/biossíntese , Neoplasias Cutâneas/genética , Proteína Supressora de Tumor p53/biossíntese , Estudos de Casos e Controles , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Melanoma/metabolismo , Melanoma/mortalidade , Melanoma/patologia , Invasividade Neoplásica , Metástase Neoplásica , Proteínas de Neoplasias/genética , Prognóstico , Estudos Retrospectivos , Risco , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/patologia , Austrália Ocidental/epidemiologiaRESUMO
Mutations of the p53 tumour suppressor gene are common to many human malignancies. Although increased p53 expression has been observed in cutaneous malignant melanoma, mutations of the p53 gene appear to be infrequent. We examined 140 benign and malignant paraffin-embedded melanocytic lesions for p53 protein expression by immunohistochemistry, using the monoclonal anti-p53 antibody DO-7 and a microwave method of antigen retrieval. Fifteen naevi and 25 melanomas were further analysed for p53 mutations within exons 5-8 of the p53 gene. DNA was extracted from paraffin sections and screening for mutations was carried out using PCR-SSCP. We demonstrated p53 protein expression in 33% of naevi (17 out of 51), 35% of primary melanomas (20 out of 58), and 70% of metastatic lesions (15 out of 21). p53 expression in benign lesions was weaker than in malignant lesions in intensity and percentage of cells staining. p53 protein expression in melanomas increased in intensity and percentage of cells staining with tumour progression. In 25% (three out of 12) of metastatic melanomas p53 mutations were detected by PCR-SSCP and increased expression of p53 protein was observed in these tumours. p53 gene mutations were not detected in any benign melanocytic lesions. We demonstrate that antigen retrieval techniques increase p53 immunoreactivity in paraffin embedded melanocytic tissues. p53 protein expression in melanomas increases with depth of tumour invasion. As p53 gene mutations occur infrequently in malignant melanoma, other mechanisms are proposed to influence p53 protein expression in melanocytic lesions.
Assuntos
Regulação Neoplásica da Expressão Gênica , Genes p53/genética , Melanoma/genética , Mutação , Nevo/genética , Neoplasias Cutâneas/genética , Sequência de Bases , DNA de Neoplasias/análise , Humanos , Imuno-Histoquímica , Melanoma/imunologia , Melanoma/secundário , Dados de Sequência Molecular , Nevo/imunologia , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Neoplasias Cutâneas/imunologia , Proteína Supressora de Tumor p53/análiseRESUMO
We report the first documented case of undifferentiated carcinoma of the renal pelvis with a prominent lymphoid stroma (lymphoepithelioma-like carcinoma [LELC]). LELCs are morphologically identical to nasopharyngeal carcinoma and are rarely seen in the urinary tract, with only isolated cases involving the urinary bladder and ureter. The tumor was composed entirely of large pale staining malignant epithelial cells with ill-defined borders arranged in syncytial sheets separated by mainly reactive lymphocytes, occasional plasma cells and histiocytes. Tumor cells were immunoreactive to cytokeratin and were negative for leukocyte common antigen. Awareness of LELC is important, as it should be distinguished from lymphoma or inflammatory lesions including, xanthogranulomatous pyelonephritis.
Assuntos
Carcinoma de Células Escamosas/patologia , Neoplasias Renais/patologia , Pelve Renal/patologia , Idoso , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/diagnóstico por imagem , Carcinoma de Células Escamosas/cirurgia , Feminino , Humanos , Imuno-Histoquímica , Queratinas/análise , Neoplasias Renais/química , Neoplasias Renais/diagnóstico por imagem , Neoplasias Renais/cirurgia , Pelve Renal/química , Pelve Renal/diagnóstico por imagem , Pelve Renal/cirurgia , Antígenos Comuns de Leucócito/análise , Tomografia Computadorizada por Raios XRESUMO
The oncogene c-erbB-2 has been shown to be amplified in 17-30% of breast cancers, with similar levels of overexpression of the oncogene product p185, a transmembrane growth factor receptor glycoprotein. Amplification of c-erbB-2 is now generally considered to be a significant prognostic indicator in patients with breast cancer. A series of 74 consecutive breast carcinomas were analysed for c-erbB-2 amplification and p185 overexpression. The procedures of Southern blotting and slot blot were used for the analysis of oncogene amplification, while immunoperoxidase (IPOX) staining and enzyme-linked immunosorbent assay (ELISA) were used for the analysis of p185 overexpression. Detection of c-erbB-2 oncogene amplification by both the conventional Southern blotting technique and by the slot blot technique showed complete accord, with the amplified c-erbB-2 oncogene being detected in 14 of the 74 patients (18.9%). The c-erbB-2 oncoprotein, as measured by IPOX and ELISA, was found to be overexpressed in 21% and 19% of patients, respectively. Comparison was made between the results attained by all four methods, and further comparison of the techniques was made from the point of view of ease of use, expense and ease of introduction into routine diagnostic laboratories. Immunocytochemistry in combination with slot blotting procedures were considered to be the most cost effective methods for evaluation of overexpression and amplification in routine pathology laboratories.
Assuntos
Southern Blotting , Neoplasias da Mama/química , Carcinoma/química , Técnicas Imunoenzimáticas , Proteínas Proto-Oncogênicas/análise , Neoplasias da Mama/genética , Carcinoma/genética , Feminino , Humanos , Receptor ErbB-2 , Sensibilidade e EspecificidadeRESUMO
Expression levels of nm23-H1 were evaluated in a variety of normal benign and malignant breast tissues by Northern and slot blot. Tissues from 153 patients presenting with palpable breast lesions were studied: 132 primary infiltrating breast cancers, 9 pure duct carcinoma in situ lesions, a phyllodes tumor, 9 benign lesions and 2 local recurrences of carcinoma. In addition to lesional tissue, 49 samples of macroscopically normal breast tissue, 37 axillary lymph nodes and 9 samples from patients undergoing cosmetic reduction mammoplasty were studied. Sets of normal breast tissue, primary tumor and lymph node tissue from individual patients were available for comparison in 37 cases. A wide range of gene expression was detected in the various tissue types. The highest levels of expression were detected in malignant samples with in situ carcinomas being associated with the highest levels of gene expression. The expression levels of nm23-H1 in normal breast tissue were lower than the corresponding tumors from the same patients (p < 0.0005). Benign breast lesions (including 6 fibroadenomas) had levels of gene expression approximating those of the normal tissue samples. Normal axillary lymph nodes had significantly lower levels of nm23-H1 expression than nodes with metastatic deposits (p < 0.03). No significant association was observed between nm23-H1 expression levels and axillary node status in patients with infiltrating carcinoma, although there was a slight trend toward lower nm23-H1 mRNA levels in the node negative group.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Doenças Mamárias/genética , Neoplasias da Mama/genética , Proteínas Monoméricas de Ligação ao GTP , Núcleosídeo-Difosfato Quinase , RNA Mensageiro/biossíntese , Fatores de Transcrição/genética , Northern Blotting , Neoplasias da Mama/patologia , Carcinoma/genética , Humanos , Metástase Linfática/genética , Nucleosídeo NM23 Difosfato Quinases , Proteínas de Neoplasias/genética , RNA Neoplásico/biossínteseRESUMO
This study examined the protection induced by oil adjuvant vaccine and broth bacterin in mice. Protective immunity was induced by both oil adjuvant and bacterin vaccination procedures. Oil adjuvant vaccination induced a 10(5)-fold increase for lethal challenge over control mice, while secondary vaccination induced a further 10-fold increase in resistance to lethal challenge. Broth bacterin induced a slightly weaker protective response with 10(4)- and 10(5)-fold increases in resistance to lethal challenge following primary and secondary vaccination, respectively. There was a significant relationship between IgG antibody levels and resistance to challenge (P = 0.026). Protection lasted for at least 20 weeks after a primary oil adjuvant vaccination. There was also a strong and significant relationship between IgG antibody levels and the passive protection afforded by serum transfer in each experiment within this study and the overall correlation was highly significant (P = 0.00001). There appeared to be a relationship between protection and the antibody response to major protein bands with the apparent molecular mass Mr. 94,000; 80,000; 67,000; 35,000 and 32,000 as well as to the bands in the region of the lipopolysaccharide components of P. multocida (approximately Mr, 14-15,000). Whether protection resulted from recognition of specific antigens or was a result of both antibody levels and antibody specificity remains to be defined.
Assuntos
Anticorpos Antibacterianos/biossíntese , Vacinas Bacterianas/imunologia , Septicemia Hemorrágica/veterinária , Pasteurella/imunologia , Adjuvantes Imunológicos , Animais , Feminino , Septicemia Hemorrágica/imunologia , Imunização Passiva , Imunização Secundária/veterinária , Immunoblotting , Imunoglobulina G/biossíntese , Imunoglobulina M/biossíntese , Cinética , Camundongos , Camundongos Endogâmicos BALB C , Vacinação/veterináriaRESUMO
Ribotyping and field alternation gel electrophoresis (FAGE) were used to examine 19 Pasteurella multocida isolates, and to assess the ability of these techniques to differentiate P. multocida strains that cause haemorrhagic septicaemia (HS). Reproducible patterns were obtained from both methods, with FAGE demonstrating greater discriminatory power than ribotyping. FAGE analysis was particularly useful in distinguishing North American cultures originating from the 1922 Yellowstone National Park Buffalo 'B' strain, demonstrating the ability to detect genetic alterations induced by repeated subculture. A remarkable homogeneity was observed among Asian HS strains following ribotyping and FAGE analysis, with a clear distinction observed between virulent and avirulent HS isolates. This study has illustrated the value of genomic fingerprinting methods in distinguishing strains of similar serotype, and the capability of these methods to produce detailed characterisation of P. multocida isolates.
Assuntos
Doenças dos Bovinos , Septicemia Hemorrágica/microbiologia , Septicemia Hemorrágica/veterinária , Infecções por Pasteurella/microbiologia , Infecções por Pasteurella/veterinária , Pasteurella multocida/classificação , Animais , Bison , Búfalos , Bovinos , DNA Bacteriano/análise , Cervos , Desoxirribonucleases de Sítio Específico do Tipo II , Eletroforese , Técnicas Genéticas , Humanos , Pasteurella multocida/isolamento & purificação , Polimorfismo de Fragmento de Restrição , Reprodutibilidade dos Testes , SorotipagemRESUMO
The oncogene c-erbB-2 is frequently amplified in human breast carcinoma. The c-erbB-2 gene is present as a single copy in normal cells, and has been mapped to chromosome 17 in the region 17q 12-21.32. c-erbB-2 encodes a transmembrane glycoprotein known as p185. The intracellular component of p185 has tyrosine kinase activity; the extracellular domain has a structure resembling a growth factor receptor. c-erbB-2 amplification, p185 overexpression and levels of transcribed c-erbB-2 specific messenger RNA have been studied in a large number of breast carcinomas using a variety of techniques. In general, overexpression of p185 oncoprotein reflects various levels of DNA amplification, though in some cases amplification can be detected in the absence of overexpression of p185 and similarly overexpression of p185 can be present without detectable levels of c-erbB-2 amplification. This findings suggests that multiple mechanisms may be responsible for overexpression. c-erbB-2 amplification and/or overexpression occurs in almost all cases of high grade duct carcinoma in-situ, but has been reported in only 10%-40% of infiltrating duct carcinoma. c-erbB-2 amplification or overexpression occurs rarely in invasive lobular carcinoma, and has not been detected in ductal or lobular epithelial hyperplasia, or in atypical ductal or atypical lobular hyperplasia. It is generally believed that c-erbB-2 amplification/overexpression is an important independent prognostic indicator in breast carcinoma, identifying a subset of patients with poor prognosis tumours, particularly if axillary node metasases are present. However, many unanswered questions remain regarding c-erbB-2 and its role in breast cancer development and progression. The causes of c-erbB-2 amplification are unknown. There is no evidence of mutations in the human gene which might cause amplification or overexpression. The significance of the differences in levels of c-erbB-2 amplification/overexpression in in-situ duct carcinoma and associated invasive duct carcinoma has not been established. Amplification or overexpression have not been reported in atypical duct hyperplasia, a proposed precursor of duct carcinoma in-situ, yet overexpression occurs almost always in high grade duct carcinoma in-situ. c-erbB-2 may play a critical role in the development of a clonal in-situ, proliferation of high histological grade, yet does not obviously influence the acquisition of an invasive phenotype. We would postulated that this instability in amplification/overexpression is of biological significance, and if better understood may aid in the study of progression of human breast carcinoma.
Assuntos
Neoplasias da Mama/genética , Receptores ErbB/biossíntese , Receptores ErbB/genética , Oncogenes/genética , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas/genética , Southern Blotting , Neoplasias da Mama/diagnóstico , Feminino , Amplificação de Genes , Humanos , Hibridização In Situ , Reação em Cadeia da Polimerase , Prognóstico , Receptor ErbB-2RESUMO
Separation of renal cell tumors into different prognostic groups is an imperative function of the diagnostic pathologist. Recently, chromophobe renal carcinoma has been described as a tumor that is morphologically distinct from conventional "clear cell" carcinoma and that has a low metastatic potential. Identification is based on routine light microscopic features and is confirmed by special stains, immunohistochemistry, and electron microscopy. We present a variant of chromophobe renal carcinoma that did not show the typical cytomorphologic features on light microscopy after formaldehyde fixation. After fixation in Solufix (a commercial fixative), these features were recognized and the diagnosis was confirmed. The tumor also showed an unusual form of calcification and psammoma body formation not previously recognized in chromophobe tumors. Molecular biological assessment was inconclusive, but excluded a chromosome 3p deletion usually found in conventional renal carcinoma. The use of a different primary fixative may provide a cost-effective screening tool to detect variant renal tumors and may have important prognostic implications.
Assuntos
Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , Segunda Neoplasia Primária/patologia , Idoso , Calcinose/patologia , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/cirurgia , Carcinoma de Células Renais/ultraestrutura , Deleção Cromossômica , Cromossomos Humanos Par 3 , Fixadores , Variação Genética , Humanos , Neoplasias Renais/genética , Neoplasias Renais/cirurgia , Neoplasias Renais/ultraestrutura , Masculino , Segunda Neoplasia Primária/genética , Segunda Neoplasia Primária/cirurgia , Nefrectomia , Prognóstico , Neoplasias da Próstata/radioterapia , Fixação de Tecidos/métodosRESUMO
Sarcomatoid renal cell carcinoma (SRCC) is an aggressive tumor variant thought to arise predominantly from dedifferentiation of clear cell carcinoma. A few reports of SRCC associated with non-clear cell tumors led to the presumption that SRCC may arise from any renal cell carcinoma, although direct evidence of this is lacking. Cytogenetic studies on 3 previously documented SRCCs associated with papillary renal cancers showed either 3p deletions or absence of trisomy 7, 17 in the sarcomatoid tumors, suggesting origin from a coexistent clear cell tumor. The present case represents the first conclusive evidence of direct progression of non-clear cell carcinoma to SRCC with both tumor components containing multiple copies of chromosomes 7 and 17. Many genetic anomalies, including p53 mutations, frequently recognized in SRCC were not recognized in this case, highlighting the importance of cytogenetic evaluation of all SRCC. The patient is well and without evidence of tumor progression 1 year after surgery, and the sinister outlook of SRCC in association with clear cell carcinoma may not apply in SRCC of non-clear cell origin.
Assuntos
Carcinoma Papilar/patologia , Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , Idoso , Carcinoma Papilar/genética , Carcinoma de Células Renais/genética , Aberrações Cromossômicas , Cromossomos Humanos Par 17/genética , Cromossomos Humanos Par 7/genética , Análise Citogenética , Feminino , Humanos , Cariotipagem , Neoplasias Renais/genética , Sarcoma/patologiaRESUMO
Amplification of multiple P multocida genomic DNA fragments by outwardly-directed primers based on the repetitive extragenic palindromic (REP) consensus sequence, generated complex profiles in a PCR-based fingerprinting method known as REP-PCR. Polymorphisms within REP-PCR profiles were used to characterise 38 isolates of P multocida. The high degree of homogeneity observed among haemorrhagic septicaemia (HS) strains of serotype B and E provided evidence of a disease-associated REP profile that may serve as a novel method for the identification of HS strains regardless of serotype. REP-PCR profiles of other P multocida serotypes were highly variable, illustrating the potential of this technique for the molecular fingerprinting of fowl cholera or atrophic rhinitis isolates. A specific amplified REP fragment was isolated and used to probe membrane-bound digested P multocida genomic DNA. Hybridisation patterns not only distinguished HS-causing isolates from non-HS P multocida, but also demonstrated a degree of relatedness between HS and HS-like strains.
Assuntos
Septicemia Hemorrágica/veterinária , Pasteurella multocida/genética , Reação em Cadeia da Polimerase/veterinária , Animais , Sequência de Bases , Doenças das Aves/etiologia , Doenças das Aves/microbiologia , Aves , Bison , Búfalos , Bovinos , Impressões Digitais de DNA/veterinária , Fragmentação do DNA , DNA Bacteriano/análise , DNA Bacteriano/química , DNA Bacteriano/genética , Cervos , Septicemia Hemorrágica/etiologia , Humanos , Pasteurella multocida/classificação , Pasteurella multocida/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Doenças das Aves Domésticas/etiologia , Doenças das Aves Domésticas/microbiologia , Suínos , Doenças dos Suínos/etiologia , Doenças dos Suínos/microbiologia , PerusRESUMO
Haemorrhagic septicaemia-causing strains of Pasteurella multocida were identified by a disease-specific ELISA. Some strains, however, were of the same serotype as those which cause haemorrhagic septicaemia (HS) but were negative when tested in the disease specific ELISA. The suspect false negative isolates were passaged in mice and retested in the HS ELISA with the same result. Immunoelectron microscopy was used to examine further these suspect HS-causing strains. Monoclonal antibodies and protein A-gold showed that the suspect negative organisms were a mixture of phenotypes with less than 10 per cent, and usually less than 2 per cent, of the population expressing HS-associated epitopes. The degree of staining on the organisms expressing the HS-epitopes was of the same intensity as the positive control organism. Expression of the HS-associated epitopes is presumably too low to allow detection in the current HS ELISA.
Assuntos
Antígenos de Bactérias/análise , Septicemia Hemorrágica/veterinária , Pasteurella/imunologia , Animais , Anticorpos Monoclonais/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos/análise , Reações Falso-Negativas , Septicemia Hemorrágica/diagnóstico , Septicemia Hemorrágica/microbiologia , Microscopia Imunoeletrônica , Pasteurella/classificação , Pasteurella/ultraestrutura , Fenótipo , Valor Preditivo dos Testes , SorotipagemRESUMO
Haemorrhagic septicaemia (HS) is caused by specific serotypes of Pasteurella multocida and is one of the major economic diseases of cattle and buffalo in South East Asia. Definitive diagnosis of the disease-causing organism with the available methods is labour intensive and not totally reliable, consequently, an ELISA system to identify P multocida organisms which cause HS was developed. One hundred and twenty-four P multocida isolates were tested, 58 were type strains and 66 were field isolates. Analysis of these strains indicated the assay had a specificity of 99 per cent and sensitivity of at least 86 per cent. The sensitivity could be an underestimate, as five isolates assumed to be false negative reactions may not all be HS-causing strains. The HS ELISA provides a rapid, simple, accurate and inexpensive diagnostic assay for identification of HS causing organisms but does not represent a new typing system for P multocida. This assay will also enable countries to assess the impact of HS more accurately.