Decreasing of p27(Kip1)and cyclin E protein levels is associated with progression from superficial into invasive bladder cancer.

The p27(Kip1)(p27) protein is a cyclin-dependent kinase inhibitor of the transition from G1 to S phase. It has been reported that decreased p27 protein level is a negative prognostic indicator in human tumours including bladder cancer. We studied the relationship between protein levels of p27, cyclin E and Ki-67 and clinicopathological features of 145 consecutive Japanese patients with transitional cell carcinoma of the bladder using immunohistochemical staining. Low protein levels of p27 were associated with low staining of cyclin E (P = 0.0302), high Ki-67 index (P = 0.0306), poorly differentiated grade (P = 0.0006), muscle invasion (P = 0.0019) and lymph node metastsis (P = 0.0002). Low staining of cyclin E and high Ki-67 index correlated with poorly differentiated grade, muscle invasion and lymph node metastsis. Cyclin E protein levels was inversely related with Ki-67 index (P = 0.0002). Kaplan-Meier plots of survival rate in patients with low versus high p27 staining showed that low protein levels of p27 were associated with a shortened disease-free and overall survival (P< 0.0001 and P< 0.0001, respectively). Similarly, low staining of cyclin E and high Ki-67 index correlated with a shortened disease-free and overall survival. On multivariate analysis using Cox proportional hazards model, low protein levels of p27 and high Ki-67 index were independent predictors of shortened disease-free (P< 0.0001, P = 0.0031, respectively), and low protein levels of p27, low staining of cyclin E and high Ki-67 index of overall survival (P = 0.0017, P = 0.0009, P = 0.0003, respectively). In superficial bladder tumours (Ta, T1; 86 patients), significant correlations were observed between low p27 staining and high Ki-67 index and early recurrence (P = 0.0048, P = 0.0178, respectively). Among the recurrenced superficial tumours (35 patients), the tumours which remained at a low stage showed high protein levels of p27 and cyclin E, and the tumours which progressed to invasive disease showed a gradual decrease in p27 and cyclin E protein levels over time. Our findings suggest that decreased protein levels of p27 and cyclin E play a role in the progression of bladder cancer and to evaluate these protein levels may be useful in management of the diseases.

Prognostic significance of p27Kip1 and Ki-67 expression in carcinoma of the renal pelvis and ureter.

OBJECTIVE: To determine the significance of p27(Kip1) (p27) for tumour behaviour and prognosis of patients with transitional cell carcinoma (TCC) of the renal pelvis and ureter. PATIENTS AND METHODS: Using immunohistochemical staining, the relationship was evaluated between p27 protein level (low < 50%, high > 50%) and the Ki-67 labelling index (low < 30%, high > 30%) and clinicopathological features of 37 consecutive Japanese patients with TCC of the renal pelvis and ureter. RESULTS: Low levels of p27 correlated with higher tumour stage (P < 0.05) and positive lymph node metastases (P < 0.05). There was no significant association between p27 staining and the grade and tumour proliferation as assessed by the Ki-67 index. A high Ki-67 index correlated with higher grade and stage (P < 0.05). Kaplan-Meier plots of survival rate in patients with low or high p27 staining showed that low levels correlated with a shorter disease-free and overall survival (P < 0.001 and P < 0.01, respectively). Similarly, patients with a high Ki-67 index had lower disease-free and overall survival than those with a low Ki-67 index (P < 0.01 and P < 0.05, respectively). The Cox proportional hazards model showed that a low level of p27 was an independent predictor of a shorter disease-free (P < 0.01) and overall survival (P < 0.05) on univariate analysis, but not of overall survival on multivariate analysis. A high Ki-67 index was an independent prognostic marker for shorter disease-free survival on univariate and multivariate analysis (P < 0.01) and for overall survival on multivariate analysis (P < 0.05). In those with a high Ki-67 index, increased p27 staining was associated with a better prognosis than decreased staining for disease-free and overall survival (log-rank test, P < 0. 01 and P < 0.05, respectively). CONCLUSIONS: The finding that a low level of p27 is associated with tumour invasion and unfavourable prognosis indicates that p27 may be a useful prognostic marker for survival in upper urinary tract cancer.

The optical activity of lysozyme crystals.

The components of the gyration tensor of the enzyme lysozyme were measured by using the HAUP method: g11 = -0.90 x 10(-5) and g33 = 1.05 x 10(-5) at 303.4 K and a wavelength of 4880 A. The optical rotatory powers along the a and c axes in the same conditions were calculated: rho a = -21.3 and rho c = -24.8 degrees cm-1. The optically active property of lysozyme is strange in that, although it contains a considerable quantity of alpha-helices (about 30%), the rotatory powers are unexpectedly small in magnitude, one order of magnitude less than those of quartz and with very large anisotropy. A conceptual consideration of this phenomenon is given. In order to assess the difference between the structures in both crystalline and solution states, the chirality index r was calculated to be 0.16. This value indicates that the structural change of lysozyme from the solution into the crystalline state is expressed by an increase of 19% in optical activity. From the NMR results [Smith et al. (1993), J. Mol. Biol. 229, 930-944], it is anticipated that the r value reflects the increased constraint in atomic motion in the side chains of exposed amino acid residues in the crystalline state.

[A surgical case of quadricuspid aortic valve associated aortic regurgitation and severe mitral regurgitation due to infective endocarditis].

We report a case of rare anomaly of quadricuspid aortic valve associated aortic regurgitation and severe mitral regurgitation due to infective endocarditis. A 50-year-old man was admitted to our hospital for fever and dyspnea. The transesophageal echocardiography showed severe aortic regurgitation due to four equal aortic cusps and severe mitral regurgitation due to infective endocarditis. At the operation, aortic valve and mitral valve were replaced with 23 mm and 29 mm SJM valves. His postoperative course was uneventful.

Small cell undifferentiated carcinoma of the urinary bladder: a cytodiagnostic case report of its variant type.

Presented is a case report of a urinary bladder carcinoma that had an unusual morphology and phenotype. A 65-year-old Japanese man complained of gross hematuria. Cytological examination of the urine before a partial cystectomy revealed small, round atypical cells with a high nuclear/cytoplasmic ratio, scant cytoplasm, and hyperchromatic nuclei with coarse and granular chromatin in a bloody background. Several tumor cells had relatively large and vesicular nuclei with prominent eosinophilic nucleoli and obscure perinucleolar halos. A small number of large atypical urothelial cells were also recognized. The tumor recurred locally 3 months after the operation. The urine cytology during recurrence showed the same features without the atypical urothelial cells. These cytological findings suggested a case of small cell undifferentiated carcinoma (SCUC) combined with transitional cell carcinoma (TCC). An histology of the resected specimen before the recurrence revealed that the SCUC was consistent with a variant type of SCUC proposed for the lung and showed transition with TCC in situ. M-VAC chemotherapy after a total cystectomy was less effective. The patient died 6 months after diagnosis. A variant subtype of SCUC of the urinary bladder associated with TCC in situ has not been previously reported. Although this histological type is very rare, its earlier cytological detection is needed for appropriate therapy.

Immunohistochemical and ultrastructural study of mixed small cell/large cell carcinoma of the lung. Expression of Sialyl LeX-i antigens and scarcity of neuroendocrine characteristics.

The authors examined the immunohistochemical and ultrastructural characteristics of mixed small cell/large cell carcinoma (mixed subtype) of the lung which has been newly categorized as anaplastic small cell lung carcinoma. The 34 cases of small cell lung carcinoma examined consisted of 18 cases of pure small cell carcinoma (pure subtype), 12 cases of the mixed subtype and 4 cases of combined small cell carcinoma. Immunohistochemically, the number of immunoreactive tumor cells for chromogranin A was smaller (P less than 0.05) and that for creatine kinase BB was relatively larger in the mixed subtype. Sialyl LeX-i antigen (SLX), one of the SSEA-1 related carbohydrate antigens, was expressed specifically in the mixed subtype. Ultrastructurally, tumor cells in the mixed subtype had smaller amounts of neurosecretory granules and larger amounts of desmosomes than those in the pure subtype (P less than 0.05, P less than 0.005, respectively). These results suggest that the mixed subtype may show more epithelial than neuroendocrine differentiation and that SLX may be a specific marker for the mixed subtype to be used in pretherapeutic diagnosis.

[A case of synchronous multiple primary cancers of the stomach and kidney].

In line with an increase in the incidence of multiple primary cancers, we have encountered a case of synchronous multiple primary cancers of the stomach and the left kidney. The patient, a 69-year-old male, visited our hospital after experiencing epigastric discomfort for three months. An advanced gastric cancer, Borrmann III type, was detected by endoscopic examination. Preoperative abdominal computed tomography also revealed a large low density mass occupying the upper part of the left kidney. On angiography, the left kidney showed a hypervascular mass, showing pooling and tumoral stains, thereby suggesting a renal cell carcinoma. The patient thus underwent a subtotal gastrectomy with an R2 lymph node dissection and a left radical nephrectomy. Histologically, the gastric lesion was a poorly-differentiated adenocarcinoma and the left renal lesion was a renal cell carcinoma of the clear cell type.

In vitro transfer of apolipoproteins from plasma lipoproteins to artificial lipid particles.

Our earlier studies in vivo revealed that artificial lipid particles (exo TG) in lipid emulsion acquire apolipoprotein C-II (apo C-II) from high density lipoprotein (HDL). Since the transfer of apo C-II to exo TG terminated within a short time after the intravenous injection of exo TG, it is likely that exo TG has a high affinity for apolipoprotein. This hypothesis has been investigated in vitro. Human plasma was incubated at 37 degrees C with or without a 10% emulsion of soybean oil for 5 min and 60 min. After the incubation, the plasma was separated into HDL, low density lipoprotein (LDL) and very low density lipoprotein (VLDL). Levels of apo C-II, C-III and E in each lipoprotein fraction were quantified. When the plasma was incubated without exo TG, the distribution of apo C-II and C-III in the lipoprotein fractions was unchanged after incubations for 5 and 60 min. However, when the plasma was incubated with exo TG, apo C-II and C-III in the HDL fraction decreased after a 5 min incubation, while those in the VLDL fraction increased. Apo E in each lipoprotein fraction did not change after 5- and 60-min incubations regardless of the presence or absence of exo TG.(ABSTRACT TRUNCATED AT 250 WORDS)

[A case of pulmonary sarcoidosis with calcification in the lung].

A 35 year-old male farmer presented with complaints of productive cough and sputum. The chest X-ray films showed reticulonodular shadows bilaterally in the upper and middle lung fields, segmental infiltration in the right lung, and no BHL. Tuberculin reaction was negative. Serum angiotensin converting enzyme level was 26.5 IU/ml. Precipitating antibodies for Thermophilic actinomycetes and M.f. were negative. BAL showed moderate lymphocytosis (24.3%), and CD 4/8 was 1.67. A biopsy specimen of right scalene lymph node showed epithelioid cell granulomas and TBLB epithelioid cell granulomas and spot-like calcification. Pulmonary sarcoidosis with calcification in the lung is very rare.

Inhibitory effects of exogenous insulin on oxidative utilization of glucose in septic rats.

The influence of exogenous insulin on oxidation of U-14C-glucose was investigated by monitoring the production of 14CO2 in septic rats. Ten Wistar rats underwent ligation of the distal third of the caecum and the ligated caecum was punctured with a 21-G needle (septic rats: group S). A group of ten rats was not subjected to these procedures (control rats: group C). Both groups received parenteral nutrition (PN) for the subsequent 27 hours. Non-protein nutrients were provided exclusively as glucose. Five rats in each group received insulin at a rate of 0.64 U/kg/hr for the last six hours of PN, and these rats were designated groups CI and SI. At the 21st hour of PN, 1.563 muCi of U-14C-glucose was injected as a bolus, and the production of 14CO2 was measured. The cumulative production of 14CO2 for the subsequent six hours, given as a percentage of the 14C injected, was 78.22 +/- 6.22% in group C, 86.16 +/- 4.23% in group CI, 62.38 +/- 13.00% in group S, and 54.38 +/- 6.89% in group SI. The elevated levels of blood glucose in rats in group S were reduced by the administration of insulin. These results indicate that the reduction of elevated blood glucose levels by exogenous insulin in the septic rats with antecedent hyperinsulinemia does not reflect an increase in the oxidative utilization of glucose but rather inhibition of the oxidation of glucose.

Influences of an infusion of lipid emulsion on phagocytotic activity of cultured Kupffer's cells in septic rats.

An experimental study was undertaken to study the influences of an infusion of lipid emulsion on phagocytosis of Kupffer's cells in septic rats. Sepsis was induced in 13 rats by ligating the cecum. Five of them received glucose as the sole nonprotein calorie (septic-glucose group), four of the rats received 25% of the nonprotein calorie with lipid emulsion, Intralipid (septic-lipid group), and the remaining four rats did not receive any intravenous solution and were allowed access to water (septic-fasted group). Another four rats which received neither intravenous solution nor ligation of the cecum served as the control group. The intravenous infusion was carried out for 72 hr. The phagocytotic activity of Kupffer's cells was determined by the ability to engulf latex particles with a size of 1.09 micron, in vitro. The phagocytotic activity was enhanced by the presence of sepsis but it was inhibited by starvation. The difference in the phagocytotic activity between the septic-glucose group and the septic-lipid group was not significant. These results suggest that, insofar as an in vitro study is concerned, a 72-hr infusion of lipid emulsion at a rate of 25% of the total nonprotein calorie does not influence the phagocytotic activity of cultured Kupffer's cell obtained from septic rats.

Effects of total parenteral nutrition on the development of intestinal endotoxemia in rats: a comparison between glucose and lipid.

The effects of parenteral nutrition (PN) and of the difference in the PN regimens between glucose and lipid emulsion on the development of endogenous endotoxemia were studied in 40 Wister rats. Endotoxemia was induced by occluding the superior mesenteric vein (SMV) for 30 min. The plasma endotoxin in the portal blood at the time of the release of the SMV occlusion and that in the arterial blood 10 min after the release were quantified. Twenty of the 40 rats had received PN for 48 hr prior to the SMV occlusion. Ten of these 20 rats received the total nonprotein calorie (TNPC) solely with glucose, and the other 10 rats received 25% of the TNPC with lipid emulsion. Ten rats had been allowed free access to lab food until the SMV occlusion. The remaining 10 rats underwent neither the SMV occlusion nor PN, and served as the control group. Both the portal and the arterial endotoxin increased after the release of the SMV occlusion, however the portal endotoxin was higher than that of the arterial one. Both the portal and the arterial endotoxin of the rats supported by PN were significantly lower than those of the rats nourished by lab food, while they were higher than the control values. The difference in the PN regimens did not cause any alteration in the endotoxin levels. These results indicate that the development of intestinal endotoxemia was not influenced by the difference in the PN regimens, but it was rather influenced by a presence of intestinal content.