Telomerase activity and expression of human telomerase RNA component and human telomerase reverse transcriptase in lung carcinomas.

The aim of this study was to evaluate the usefulness of determination of telomerase activity and expression of human telomerase RNA component (hTERC) and human telomerase reverse transcriptase (hTERT) for the diagnosis of lung carcinomas. The tissues studied consisted of 115 carcinomas and adjacent nonneoplastic lung, which were removed surgically without previous chemotherapy or radiotherapy. Telomerase activity was determined using a semiquantitative polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay. The results obtained were classified into high and low telomerase groups. Localization of expression was examined by using in situ hybridization and immunohistochemistry. The correlation between telomerase activity in lung carcinoma and clinicopathologic features, including prognosis, was investigated. Telomerase activity in lung carcinomas was detected in 107 of 115 (93%) lung carcinomas, but not in any adjacent noncancerous tissues, and was significantly higher in small cell carcinoma than in any other histologic type. This activity also was significantly higher in poorly differentiated than in well-differentiated squamous cell carcinomas and adenocarcinomas. The overall survival rate (P =.020) was significantly lower in the high telomerase group. Messenger RNAs for hTERC and hTERT were mainly detected in the cytoplasm of cancer cells by in situ hybridization, and TERT protein was localized in the nuclei of these cells by immunohistochemical staining. Determinations of telomerase activity by in situ hybridization, immunohistochemistry, and TRAP assay are useful for evaluating the diagnosis and prognosis of lung carcinomas.

Defining a common region of DNA amplification at 22q11.2-12 in head and neck squamous cell carcinomas by quantitative FISH analysis.

By comparative genomic hybridization (CGH) with 17 head and neck squamous cell carcinoma (HNSCC) cell lines, we previously detected an amplified region as a distinct peak at 22q11.2-12 in 3 cell lines. Because the possible presence of an oncogene was strongly suggested, the region was mapped in more detail by defining the minimum region that was commonly amplified by using fluorescence in situ hybridization (FISH) with a series of cosmids. Eighteen cosmid clones on 22q11.2-12 were assigned to their locations as a fractional length map and hybridized to cells from three HNSCC cell lines. The three cosmid clones, cHKA-118, cHKAD-26, and D22S938, showed the highest levels of amplification, and the size of the amplicon was calculated to be approximately 1.7 Mb in the OM1 and HSC6 cell lines. Several genes related to oncogenesis, including PRKM1, map to this locus. Thus, the definition of the common region with the highest level of copy number increases by FISH provides a starting point for identifying the gene that may play an important role in the development of HNSCC.

Mucin histochemistry in primary adenocarcinoma of the urinary bladder (of urachal or vesicular origin) and metastatic adenocarcinoma originating in the colorectum.

In order to evaluate the mucin histochemistry of primary adenocarcinomas (PA) of the urinary bladder and metastatic adenocarcinoma (MA) originating in the colorectum, 52 PA and nine MA were examined. It was determined that the percentage of cases in which more than 25% of the tumor was stained by each of the following: (i) Alcian blue pH 2.5 periodic acid-Schiff (AB-PAS); (ii) high iron diamine-AB (HID-AB); (iii) periodic acid-sodium borohydride-potassium hydroxide-PAS (PA-SB-PH-PAS); (iv) galactose oxidase- Schiff (GOS); and (v) paradoxical concanavalin A stain (PCS). For PA, the values obtained were: 75% of cases (blue, AB-PAS), 85% (magenta, AB-PAS), 71% (black, HID-AB), 75% (blue, HID-AB), 0% (PA-SB-PH-PAS), 19% (GOS), 8% (class II concanavalin A (Con A)-reactive mucin)), and 0% (class III Con A-reactive mucin). For MA, the corresponding values were 33, 22, 0, 11, 0, 0, 11, and 0%, respectively. A higher percentage of PA than MA cases showed staining in AB-PAS for acidic and neutral mucins, in HID-AB for sialo- and sulfomucins, and in GOS for terminal beta-galactose and beta-N-acetylgalactosamine. PA and MA were significantly different in terms of both frequency of staining with AB-PAS and frequency of staining with HID-AB. However, the overlap was such that in practice, it might be difficult, if not impossible, to use mucin histochemistry to inform a differential diagnosis. In view of the differences in AB-PAS and HID-AB positivity between PA and MA, we speculate that MA (originating in the colorectum) may have undergone structural distortion affecting the production and/or secretion of neutral mucins and acidic mucins (sialo- and sulfomucins) during metastasis or invasion.

Studies of glomerular permeability factor (GPF) in focal segmental glomerular sclerosis and the relationship between GPF and vascular permeability factor (VPF).

BACKGROUND: We previously demonstrated that the supernatants of cultured concanavalin-A (con-A) stimulated peripheral blood mononuclear cells (PBMC) from patients with minimal change nephrotic syndrome (MCNS) increased the urinary protein excretion in injected rats and suggested that PBMC released a factor, which we called glomerular permeability factor (GPF), changes in the glomerular permeability and thus resulted in proteinuria in MCNS. MATERIAL AND METHODS: In this study we investigated the GPF activity in focal segmental glomerular sclerosis (FGS) and other conditions of chronic glomerulonephritis (CGN), and also the relationship between GPF and vascular permeability factor (VPF). In experiment 1 the supernatants of the cultured con-A stimulated PBMC from patients with 10 FGS, 5 other CGN and 10 controls were tested regarding their ability to produce GPE The GPF activity was defined as positive when the 8-hour urinary protein excretion after the injection of the supernatant in Sprague-Dawley rats exceeded the mean value plus 2 standard deviations (M + 2 SD) of that before injection. RESULTS: Three out of 10 FGS patients and 1 membranous nephropathy patient out of the 5 other CGN patients were positive for GPF activity. In experiment 2 the relationship between GPF and VPF was analyzed using culture supernatants of PBMC from 10 nephrotic MCNS patients and 15 controls. The VPF activity was measured following the method developed by Ovary [1975]. All 7 cases that were positive for GPF activity were simultaneously positive for VPF activity. On the other hand, 16 cases that were positive for VPF activity were not always positive for GPF activity (7 cases were positive and 9 were negative for VPF activity). CONCLUSION: Experiments 1 and 2 thus suggested that GPF was not active in MCNS alone, but also in other CGN conditions and it was therefore not considered to be the same factor/substance(s) as VPF.

Expression of C-type natriuretic peptide during development of rat lung.

C-type natriuretic peptide (CNP), recently found to be secreted from vascular endothelial cells, is now viewed as a novel endothelium-derived relaxing peptide. However, the distribution and expression of CNP during cardiopulmonary development is unclear. To follow changes in the expression of CNP during lung development, we examined rat embryos and neonates using Northern blot analysis and in situ hybridization for CNP mRNA and radioimmunoassay and immunohistochemistry for CNP protein. A substantial expression of CNP mRNA was first detected on postnatal day 2, and it thereafter remained fairly steady. The level of CNP protein also increased rapidly after postnatal day 1, reaching a settled level on postnatal day 4. CNP protein and mRNA were detected in the endothelium and smooth muscle cells of blood vessels and in bronchial airway and alveolar epithelia. Immunoreactivity for CNP protein in the endothelium of blood vessels increased to an intense level after the saccular stage. These results suggest that the changes in CNP levels may be related to the occurrence of pulmonary vasodilation after birth.

Expression of telomerase mRNA component (hTR) in transitional cell carcinoma of the upper urinary tract.

BACKGROUND: Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric repeats onto chromosomal ends using a segment of its RNA component as a template. Its activity has become an established indicator of the diagnosis, biologic behavior, and prognosis of several tumors. However, to the authors' knowledge, no previous study has investigated the diagnostic and prognostic importance of the expression of telomerase RNA component (hTR) in transitional cell carcinoma of the upper urinary tract (TCC-UUT). METHODS: The authors investigated hTR expression using in situ hybridization in 130 cases of TCC-UUT, and also its relation to proliferating cell nuclear antigen (PCNA) immunoreactivity, immunoreactivity for p53 oncoprotein, clinicopathologic parameters, and clinical outcome. RESULTS: Positive hTR expression was recognized in 98.4% of the samples and was apparent within the cytoplasm of tumor cells. In normal urothelium, its expression was restricted to the basal cell layers, whereas in the dysplastic lesions of TCC-UUT it was detected with the same intensity and distribution as in the tumor itself. No correlation was found between hTR expression and clinicopathologic findings, PCNA index, or the expression of p53 oncoprotein, although hTR score did tend to increase with disease stage. In univariate and multivariate analyses of disease free and overall survival rates, high hTR expression was associated with significant decreases in both rates. CONCLUSIONS: The expression of hTR appears to be a useful indicator of prognosis for patients with TCC-UUT. In addition, evidence of up-regulation of hTR may also be useful in the diagnosis of this disease.

Expression of endothelin-1 in rats developing hypobaric hypoxia-induced pulmonary hypertension.

Experimental pulmonary hypertension induced in a hypobaric hypoxic environment (HHE) is characterized by structural remodeling of the heart and pulmonary arteries. Endothelin-1 (ET-1), a 21-amino acid peptide, is a novel and long-lasting vasoconstrictor that increases pulmonary arterial pressure in both in vivo and in vitro experiments. To study the effects of HHE on ET-1 activity in the lungs, 59 male rats were subjected to the equivalent of an altitude of 5500 m for 1 to 4 weeks. In rats exposed to HHE, the mean pulmonary arterial pressure increased significantly from 15.2+/-0.3 (ground level) to 30.6+/-1.5 mm Hg (5500-m level) at 4 weeks, whereas their mean systemic arterial pressure remained normal. The levels of ET-1 mRNA and protein, measured respectively by Northern blot analysis and enzyme immunoassay, increased rapidly in the lungs on exposure to HHE. By in situ hybridization and immunohistochemistry, respectively, ET-1 mRNA and protein were detected in control rats in nonciliated bronchiolar epithelial cells and alveolar epithelial cells, as well as in the endothelial cells of pulmonary arteries, but minimally in the smooth muscle cells of pulmonary arteries. ET-1 mRNA- and protein-reactive smooth muscle cells in pulmonary arteries and ET-1 mRNA-reactive airway epithelial cells were significantly more abundant in rats exposed to HHE than in ground level controls. These results suggest the possibility that in smooth muscle cells in pulmonary arteries and airway epithelial cells, ET-1 may play an autocrine or paracrine role in the remodeling of blood vessels during the development of the pulmonary hypertension that is induced by HHE.

Thrombopoietin expression in normal and hypobaric hypoxia-induced thrombocytopenic rats.

Thrombopoietin (TPO) is important as the physiologic regulator of platelet production. High-altitude hypoxia is a well-known cause of polycythemia and thrombocytopenia in animals. Fifty-two Wistar rats were housed for 0.5 to 21 days in a mechanical chamber in an environment equivalent to that found at 5500 m to determine (a) the cellular localization of TPO and (b) whether the decreased platelet and megakaryocyte counts in rats exposed to a hypobaric hypoxic environment (HHE) are associated with an altered TPO mRNA expression. In normal rats, there were high levels of TPO mRNA in the liver and kidney, intermediate levels in the brain and large intestine, and low levels in the skeletal muscle and small intestine. TPO mRNA and protein were expressed in Purkinje cells and neuronal cells in the brain, in proximal tubular cells and the mesangial cells of the glomeruli in the kidney, in hepatocytes and biliary duct epithelial cells, in absorptive epithelial cells in the large intestine, in the epidermis, and in the lung. The platelet count in the blood and megakaryocyte counts in the bone marrow and spleen were all decreased significantly after 5 or more days of exposure to HHE. In major producers such as the liver and kidney and in minor producers such as the brain, TPO mRNA levels, which tended to be decreased after 0.5 to 3 days of exposure to HHE, had returned to normal by about Day 5 or 7. Thus, during the HHE period with a decreased platelet count, no changes in TPO mRNA levels were detected in these three organs. In conclusion, we have demonstrated that TPO production occurs in various types of cells. In HHE, however, factors other than TPO may be involved in hypobaric hypoxia-induced thrombocytopenia in rats.

Tissue factor is associated with the nonbacterial thrombotic endocarditis induced by a hypobaric hypoxic environment in rats.

High-altitude hypoxia causes a hypercoagulable state. In our previous study on the blood coagulation system in rats, nonbacterial thrombotic endocarditis (NBTE) developed after 4-12 weeks' exposure to the equivalent of 5500 m in altitude. We hypothesized that TF (tissue factor)-producing cells in the cardiac valves might be induced by the hypobaric hypoxic environment (HHE) and then trigger NBTE. A total of 170 male Wistar rats were housed in a chamber at the equivalent of 5500 m altitude for 1-12 weeks. We measured TF activity in the plasma and studied morphological changes in the mitral valves using immunohistochemical and immunoelectrical methods for TF protein and in situ hybridization for TF mRNA. After 4 weeks or more of exposure to HHE, 28 of the 56 surviving rats had developed NBTE. After 4-8 weeks' exposure to HHE, the plasma TF activity level was significantly higher than in control rats. There was a significant correlation between plasma TF activity and the incidence of NBTE. After 1 weeks' exposure to HHE, immunoreactivity for TF protein was detected in foamy macrophages and stromal cells in the cardiac valves. In rats with NBTE, TF protein was present in foamy macrophages and spindle stromal cells and focally present in the extracellular matrix. TF mRNA was detected in some foamy macrophages within the thrombus, TF protein was localized to the rough endoplasmic reticulum and plasma membrane of many macrophages, some fibroblasts, and a few endocardial cells. TF is associated with the pathogenesis of the NBTE induced by exposure to HHE. The accumulation of TF-producing macrophages during exposure to HHE may be responsible for initiating thrombus formation.

Expression of bcl-2 oncoprotein in transitional cell carcinoma of the upper urinary tract.

We investigated the immunoreactivity for bcl-2 oncoprotein in 154 cases of transitional cell carcinoma of the upper urinary tract (TCC-UUT) and its relation with the immunoreactivity for p53 oncoprotein and proliferating cell nuclear antigen (PCNA) immunoreactivity. Immunohistochemically, bcl-2 oncoprotein was recognized as positive in 29.2% of the samples. The immunoreactivity for bcl-2 oncoprotein was significantly (P < 0.05) correlated only with stage, though there was a borderline correlation (P = 0.050) with PCNA immunoreactivity. Furthermore, in invasive TCC the immunoreactivity for bcl-2 oncoprotein was associated with PCNA immunoreactivity (P < 0.041). The 5-year disease-free and overall survival rates were 55.7% and 71.5%, respectively. A univariate analysis of survival revealed that stage, grade, pattern of growth, immunoreactivity for p53 oncoprotein, and PCNA immunoreactivity each had a significant effect on disease-free and overall survival rates, whereas the immunoreactivity for bcl-2 oncoprotein had no significant effect on either rate. In the final models of the multivariate analysis, stage was found to be the only prognostic factor for disease-free survival and for overall survival. Detection of immunoreactivity for bcl-2 oncoprotein appears to be of no real value in deciding the prognosis of TCC-UUT.

Argyrophilic nucleolar-organizer region counts and DNA status in bronchioloalveolar epithelial hyperplasia and adenocarcinoma of the lung.

Few studies have investigated more than one cell-biological parameter in bronchioloalveolar epithelial hyperplasia (BEH) and bronchioloalveolar carcinoma (BAC). The authors have examined argyrophilic nucleolar-organizer regions (AgNORs) and DNA status in surgically resected formalin-fixed paraffin-embedded specimens, 27 BEH and 62 well-differentiated adenocarcinomas, including 30 BAC. The authors measured the mean AgNOR count in 200 nuclei from sections of these regions. The authors also quantified DNA distribution in more than 200 cancer cells from sections of these regions, stained with 4',6-diamidino-2-phenylindole, using a microspectrophotometer. Fourteen lesions were interpreted as atypical BEH. The mean number of AgNORs per nucleus in BEH was 1.25 to 2.63. The mean number of AgNORs was significantly lower in both typical and atypical BEH than in either the bronchial surface epithelial type or the bronchial gland cell type of well-differentiated adenocarcinoma (P < .05). The mean number of AgNORs in atypical BEH was intermediate between that in typical BEH and that in BAC. Quantitative DNA image analysis showed DNA aneuploidy in 2 of 18 BEHs and 18 of 52 well-differentiated adenocarcinomas. The incidence of DNA aneuploidy increased in this order: typical BEH (0%, none out of 10 lesions) through atypical BEH (25.0%, 2 out of 8 lesions), to adenocarcinoma (34.6%, 18 out of 52 cases). Thus, the incidence of DNA aneuploidy in atypical BEH (25.0%) was intermediate between typical BEH (0%) and BAC (30.0%). These results suggest that atypical BEH may be closely related to BAC.

Urokinase-type plasminogen activator, its inhibitor, and its receptor in patients with upper urinary tract carcinoma.

BACKGROUND: Urokinase-type plasminogen activator (uPA) is a serine protease involved in tumor invasion and metastasis. Its activity during metastasis may be regulated by a plasminogen activator inhibitor (PAI). Furthermore, uPA exerts its action by binding to a membrane-bound receptor (uPAR). The authors attempted to examine the immunohistochemical expression of uPA, uPAR, and PAI-1 in patients with transitional cell carcinoma of the upper urinary tract (TCC-UUT). METHODS: Formalin fixed, paraffin embedded tumor tissues from 154 patients were analyzed using immunohistochemical staining. RESULTS: There was moderate to strong cytoplasmic staining for uPA, PAI-1, and uPAR in 57.8%, 96.1%, and 88.3%, respectively, of tumor epithelial cells, and in 22.7%, 53.9%, and 24.7%, respectively, of stromal cells at the tumor/stroma interface. Examination of the relationship between immunoreactive score and clinicopathologic findings revealed that the uPA score for stromal cells significantly correlated with the stage and pattern of growth of the tumors. The PAI-1 score for tumor epithelial cells and the uPAR score for stromal cells both correlated with stage, grade, and pattern of growth. The PAI-1-score for stromal cells correlated with stage and grade. The uPAR-score for tumor epithelial cells correlated with stage. When only the immunoreactive scores that were classified as "high" (if the score was > or = 5 or > or = 1, for tumor epithelial and stromal cells, respectively) were considered, univariate analysis revealed that a "high" PAI-1 score for tumor epithelial cells and a "high" uPAR score for stromal cells both were significantly associated with poor disease free and overall survivals, particularly early period survival. In the final models of the multivariate analysis, only stage (all periods, disease free survival and overall survival), and grade (12 months, overall survival) were found to be progressive or prognostic factors. CONCLUSIONS: Detection of immunoreactivity for plasminogen activator parameters appears to be of little or no value in determining the prognosis of patients with TCC-UUT, although some parameters were found to be associated with high stage or high grade of the tumors.

Expression in lung carcinomas of platelet-derived growth factor and its receptors.

Platelet-derived growth factor (PDGF) is synthesized and secreted by mesenchymal cells. We used immunohistochemistry and in situ hybridization to determine whether immunoreactivity for PDGF and PDGF receptor (PDGF-R) might be a prognostic indicator in lung carcinoma. We compared these results with those of immunohistochemistry for anti-proliferating cell nuclear antigen (anti-PCNA). Indirect immunohistochemistry and in situ hybridization were performed for PDGF B-chain, PDGF-R beta and PCNA antibodies, and PDGF B mRNA on frozen, paraffin-embedded sections of 92 surgically resected lung carcinomas (39 squamous cell carcinomas, 47 adenocarcinomas, 2 large-cell carcinomas, 2 adenosquamous carcinomas, and 2 double carcinomas). Clinicopathologic data (sex, age, stage, survival period, histologic type, and degree of cell differentiation) were evaluated using a statistical analysis system. PDGF reactivity was positive in tumor cell cytoplasm in some cases of squamous cell carcinoma (64%) and adenocarcinoma (55%) and in all cases of large-cell carcinoma, adenosquamous carcinoma, and double carcinoma. PDGF-R reactivity was detected only in tumor stroma. Positive PDGF staining was associated with a poor prognosis in patients with lung carcinoma, independent of age, sex, stage, and degree of cell differentiation (risk ratio = 2.53, p = 0.03). PDGF B mRNA was detected in 100% of PDGF-positive squamous cell carcinomas and in 85% of adenocarcinomas. There was no correlation between PDGF expression and PCNA index in lung carcinomas. Together, these results suggest that immunohistochemistry for PDGF B-chain may predict the outcome for lung carcinoma patients.

Displacement of gold marker in immunoelectron microscopy of human respiratory cilia.

Preembedding immunogold electron microscopy was performed to evaluate the position of outer arm dynein heavy chains in normal human respiratory cilia. Anti-dynein antibody (AD2), which is specific for sea urchin sperm flagellar dynein heavy chains, was used as primary antibody. Direct cross-sections of cilia were selected, and the distance between the center of a cilium and the center of a colloidal gold particle attached to the cilium (X) was measured. The distance between the center of a cilium and the farthest edge of an outer dynein arm of the cilium was measured by ordinary electron microscopy (Yo) and by immunoelectron microscopy (Yi). X was significantly longer than Yo and Yi. If it is assumed that the structure of respiratory cilia is dense and that antibodies are located at the outer side of the actual position of the heavy chains, then the average distance difference of approximately 90-120 A may represent the length of two conjugated antibodies. This length should be kept in mind when performing immunoelectron microscopy. The data suggest that AD2 recognizes the outer arm dynein heavy chains of normal human respiratory cilia.

E-cadherin expression in upper-urinary-tract carcinoma.

Carcinoma of the upper urinary tract is a relatively rare neoplasm, and few studies have dealt with clinicopathological findings and prognosis in a large number of cases. The purpose of our investigation was to look for a possible relation between E-cadherin (E-CD) immunoreactivity and clinicopathologic findings or clinical outcome in transitional-cell carcinoma of the upper urinary tract (TCC-UUT). To this end, we investigated E-CD immunoreactivity in 154 cases of TCC-UUT. E-CD immunoreactivity was recognized as being of "normal" pattern in 29.2% of samples. The relationship between E-CD immunoreactivity and clinicopathologic findings was significant for stage, grade and pattern of growth. The 5-year disease-free rate for 147 cases and 5-year overall survival rate for 154 cases were 55.7% and 71.5%, respectively. A univariate analysis of survival revealed that stage, grade, pattern of growth and E-CD immunoreactivity all had a significant effect on disease-free and overall survival rates. In the final models of multivariate analysis, however, we found that, for disease-free survival and for overall survival, only stage was a factor for progression or prognosis. Detection of E-CD immunoreactivity appears to be of limited value in deciding the prognosis of TCC-UUT.

Expression of platelet-derived growth-factor B-chain mRNA and tumor angiogenesis in invasive transitional cell carcinoma of the upper urinary tract.

In several types of carcinoma, angiogenesis is associated with metastasis and might be a prognostic indicator. Platelet-derived growth factor (PDGF) is one of several growth factors known; it is involved in the regulation of cell migration and proliferation. There have been few reports, however, dealing with the expression of PDGF B-chain mRNA and angiogenesis in transitional cell carcinoma. We evaluated both the expression of PDGF B-chain mRNA and angiogenesis in transitional cell carcinoma of the upper urinary tract and examined their interrelation with metastasis, patient prognosis, and other established clinicopathologic parameters. For this purpose, formalin-fixed, paraffin-embedded tumor tissues from 91 patients with invasive tumors were analyzed with in situ hybridization for PDGF B-chain mRNA and immunohistochemical staining of Factor VIII-related antigen to assess angiogenesis. The expression of PDGF B-chain mRNA was positive in 59 (66%) of the 91 patients. Microvessel density was significantly increased in positive PDGF B-chain mRNA cases (P < .0001). There was no significance, however, in any relationship between the expression of PDGF B-chain mRNA or between the microvessel density and the clinicopathologic findings, metastasis, or prognosis. Assessment of the expression of PDGF B-chain mRNA and microvessel density seems to be of no real value in predicting either the risk of metastasis or the prognosis in transitional cell carcinoma of the upper urinary tract.

Immunoreaction at 43 kDa with anti-ubiquitin antibody in breast neoplasms.

Protein ubiquitination has been implicated in ATP-dependent protein turnover and normal cell proliferation. To investigate whether the ubiquitin-mediated system is functionally involved in the cancerous state, we examined changes in protein ubiquitination in 52 surgically resected primary breast tumors. Immunohistochemically, ubiquitin (Ub) was identified in the cytoplasm of cancer cells, which were stained more strongly than adjacent normal ductal epithelium. Corresponding immunoblot analysis of normal and neoplastic regions of human breast showed that the immunoreaction for Ub at about 43 kDa was increased in all of the tumors (100%), regardless of the clinical stage or histologic grade. This protein, which gave a single spot on two-dimensional gel electrophoresis, had partial amino acid sequences which were identical to those of actin family members. Our results suggest that ubiquitination of this 43-kDa protein may be involved in the carcinogenesis or biological characteristics of human breast neoplasms.

Hypercoagulable state in a hypobaric, hypoxic environment causes non-bacterial thrombotic endocarditis in rats.

High-altitude hypoxia causes polycythaemia and a hypercoagulable state in humans and animals. This study examines the effects of a hypobaric, hypoxic environment (HHE) on the blood coagulation system in rats. A total of 170 male Wistar rats were housed in a chamber at the equivalent of 5500 m in altitude for 1-12 weeks. After 2 weeks of exposure to HHE, platelet counts decreased significantly; after 4 weeks, the prothrombin and activated partial thromboplastin times were significantly prolonged, compared with those of control rats. In addition, individual coagulation factors (VII, IX, X, XI, and XII) were significantly decreased at 8 weeks (P < 0.05). Levels of anti-thrombin III and alpha 2-plasmin inhibitor also decreased (between 4 and 8 weeks). After 4-12 weeks of exposure to HHE, 30 of 56 rats (54 per cent) developed (i) non-bacterial thrombotic endocarditis (NBTE) or (ii) infarction of the myocardium or kidney, or both (i) and (ii). The incidence of NBTE increased from 33 per cent (5/15 rats) at 4 weeks to 100 per cent (7/7 rats) at 12 weeks. Electron microscopy showed detached endothelial cells in the mitral valves at 1 week; platelets adhered to the subendocardial matrix and platelet aggregation with thrombus formation was seen at 2 weeks of exposure. The results suggest that exposure to HHE induces a hypercoagulable state and causes an NBTE in rats that may result in consumption coagulopathy.

Fatal diffuse alveolar damage from bird fanciers' lung.

A 68-year-old man, who had continuing exposure to budgerigars, developed fatal acute respiratory failure following years of slowly progressive pulmonary deterioration. His lung function was characterized first by mild airflow obstruction and later by progressive loss of lung volume. Computed tomography showed progressive development of pulmonary fibrosis and honeycombing. His serum disclosed precipitins to pigeon antigen. During his final illness his chest radiograph showed widespread patchy consolidation. At autopsy, his lungs revealed left lower lobe bronchopneumonia, fibrosis and honeycombing at the bases and widespread evidence of diffuse alveolar damage with organized exudate in some alveoli. To our knowledge, this is the second reported fatality due to acute alveolar injury in bird fanciers' lung.

Immunohistochemical evaluation of p53 oncoprotein in transitional cell carcinoma of the upper urinary tract.

The p53 gene, which is located on human chromosome 17, encodes for a nuclear phosphoprotein and is thought to regulate cell growth and proliferation. Although the immunoreactivity for p53 oncoprotein in transitional cell carcinoma (TCC) of the urinary bladder has been shown to correlate with clinicopathologic findings and prognoses, there have been no such reports on TCC of the upper urinary tract (TCC-UUT). The present study investigated the prognostic value of p53 oncoprotein in TCC-UUT. Formalin-fixed, paraffin-embedded tumor tissues from 149 TCC-UUT patients were analyzed using immunohistochemical staining. Immunohistochemically, p53 oncoprotein was recognized as positive in 26.8% of the samples. The immunoreactivity for p53 oncoprotein was significantly (P < .05) correlated with both stage, grade, and pattern of growth. The 5-year disease-free and overall survival rates were 58.4% and 69.7%, respectively. A univariate analysis of survival showed that stage, grade, pattern of growth, and the immunoreactivity for p53 oncoprotein have a significant effect on disease-free and overall survival rates. In the final models of multivariate analysis, only stage for disease-free survival, and stage and the immunoreactivity for p53 oncoprotein for overall survival were found to be progressive or prognostic factors. Detection of immunoreactivity for p53 oncoprotein appears to be of real value in deciding the prognosis of TCC-UUT.