P21WAF1, P27KIP1, TP53 and C-MYC analysis in 204 ovarian carcinomas treated with platinum-based regimens.

BACKGROUND: The prognostic and predictive value of cell cycle regulatory proteins in ovarian cancer has not been established. We evaluated the clinical and biological significance of P21(WAF1), P27(KIP1), C-MYC, TP53 and Ki67 expressions in ovarian cancer patients. MATERIALS AND METHODS: Immunohistochemical analysis was performed on 204 ovarian carcinomas of International Federation of Gynecology and Obstetrics (FIGO) stage IIB to IV treated with platinum-based chemotherapy. Multivariate analysis with Cox and logistic regression models was performed in the whole group, and in the TP53-negative and TP53-positive subgroups. RESULTS: High P21(WAF1) labeling index (LI) was an independent positive predictor of platinum-sensitive response (P = 0.02). Overall survival was positively influenced by P21(WAF1) LI (P = 0.02) or by P21(WAF1) plus P27(KIP1) LI (P = 0.004) in the TP53-negative group only. Ki67 LI showed borderline association with disease-free survival (P = 0.05). Growth fraction was negatively associated with P21(WAF1) and P27(KIP1) indices in the TP53-negative group (P = 0.023 and 0.008, respectively), and these associations were borderline or lost in the TP53-positive group. Endometrioid and clear cell carcinomas differed from other carcinomas by having a low incidence of TP53 accumulation, a high incidence of C-MYC overexpression (70%) and a low median Ki67 LI (all with P <0.001). CONCLUSIONS: We have shown an independent predictive value of P21(WAF1) LI in ovarian carcinoma patients. The prognostic value of P21(WAF1) and P21(WAF1) plus P27(KIP1) LI was determined by TP53 status. A high frequency of C-MYC overexpression in endometrioid and clear cell carcinomas may suggest its role in the development of these tumor types.

Evaluation of clinical significance of TP53, BCL-2, BAX and MEK1 expression in 229 ovarian carcinomas treated with platinum-based regimen.

In cell line studies, BCL-2, BAX, as well as novel MEK1 protein levels have strong influence on ovarian cancer response to cisplatin-based chemotherapy. However, such associations have not been demonstrated clinically. We evaluated prognostic/predictive significance of these proteins with regard to TP53 status. Immunohistochemical analysis was performed on 229 ovarian carcinomas FIGO stage IIB-IV treated with platinum-based chemotherapy; the results were analysed by the Cox and logistic regression models. Clinical parameters (residual tumour size, patient age, FIGO stage) were the only indicators of overall survival (OS) and the strongest predictors of complete remission (CR). On the other hand, BAX expression was the strongest (P=0.005) or the only (in FIGO IIIC, P=0.02) prognostic indicator of disease-free survival (DFS) in the TP53(+) group. TP53(+) and TP53(-) ovarian carcinomas differed in clinical and molecular prognostic and predictive factors. Another novel finding is that CR was negatively influenced by high BAX expression in all patients group (P=0.047) and by BCL2 expression in the TP53(-) group (P=0.05). High MEK1 expression was associated with endometrioid and clear cell carcinomas (P=0.049); its loss was found with advancing FIGO stage (P=0.002). Our results suggest that binomial TP53 status divides ovarian carcinomas into two biologically distinct groups. BAX expression is an important factor of DFS in the TP53(+) group. BCL-2 and BAX, but not MEK1 expressions have predictive value in ovarian cancer patients treated with platinum-based chemotherapy.

Germ cells with nuclear DNA fragmentation related to apoptotic cells in rat testis in experimental hyperprolactinemia induced by metoclopramide.

The cells with nuclear DNA fragmentation related to apoptosis were detected by TUNEL technique in the seminiferous epithelium of control rats and of rats with experimental hyperprolactinemia induced by metoclopramide. The percentage of convoluted tubules with apoptotic cells and the number of apoptotic cells (predominantly spermatogonia and spermatocytes) was increased in the experimental group. The results indicated stage-specific germ cell apoptosis. In the experimental group, apoptotic cells were most evident at early (I-IV), middle (VII-VIII) and late (XII-XIV) stages of the seminiferous epithelium cycle, as revealed by light and electron microscopy. We suggest that a decreased concentration of testosterone and an increased concentration of prolactin could disturb spermatogenesis and contribute to the intensive apoptosis of germ cells in rats with hyperprolactinemia. Sertoli cells which have receptors for testosterone and prolactin and play an important role in spermatogenesis and in the initiation of apoptosis in seminiferous epithelium, could mediate such an influence of both hormones.

Two-year follow-up after acute thromboembolic limb ischaemia: the importance of anticoagulation.

BACKGROUND: evidence on the effectiveness and usage of long-term anticoagulant therapy after acute thromboembolic limb ischaemia is very sparse. This study correlated medical events with administration of warfarin. METHOD: during a three-month audit in 1996, 287 patients with embolism or thrombosisin situ survived for 30 days, and 214 (75%) were reviewed by questionnaires returned from clinicians throughout the United Kingdom. Minimum follow-up was two years. RESULTS: thirty-five per cent had died. Recurrent acute limb ischaemia was reported in 11%, arterial intervention in 11%, and major amputation in 12%. Warfarin was given initially to 57% patients, but at follow-up only 43% were still taking warfarin (p<0. 05); reasons for stopping anticoagulation were often unknown. Recurrent limb ischaemia was less common in patients given warfarin initially (7% versus 17%) and still taking warfarin (3% versus 19%) -p;<0.05. Amputation was also less common in patients given warfarin initially (5% versus 21%) and still on warfarin (3% versus 21%) -p;<0.05. CONCLUSION: long-term oral anticoagulation was associated with reduced risk of recurrent limb ischaemia and amputation, but more research is needed to define the benefits and risks, especially for thrombosisin situ. Clinicians should give clear advice about anticoagulation when patients are discharged from hospital.

Spontaneous apoptosis in ovarian carcinomas: a positive association with p53 gene mutation is dependent on growth fraction.

Changes in cell survival contribute to tumour development, influence tumour biology and its response to chemotherapy. p53 gene alterations should negatively affect apoptosis by impaired p53-dependent apoptotic response. We looked for associations between spontaneous apoptosis, p53 gene mutation, p53 protein accumulation, growth fraction, bcl-2 expression and histological parameters in 64 ovarian, four tubal and three peritoneal carcinomas. Apoptotic cells were detected with the TUNEL method. p53 gene variants were detected by the single-strand conformation polymorphism and were sequenced directly. P53, Ki-67 and bcl-2 protein expressions were detected immunohistochemically. A weighed multiple logistic regression model was applied. Apoptotic index (AI) ranged 0.02-0.18 (mean 0.11); proliferation index (PI) ranged 3-90% (mean 54%). p53 gene mutations were present in 51, p53 protein accumulation in 46, and diffuse bcl-2 expression in 29 of 71 tumours. The AI was positively associated with the presence of p53 gene mutation (P = 0.011). However, the PI included into the analysis did positively influence the AI (P = 0.02) and diminished the association with p53 gene mutation (P = 0.082). The AI was negatively associated with good histological differentiation (P = 0.0006), the serous tumour type (P = 0.002), and diffuse bcl-2 expression (P = 0.025). Strong bcl-2 expression was associated with endometrioid tumour type (P = 0.002). FIGO stage and p53 protein accumulation were the only parameters that influenced overall survival time. Thus, our results suggest that histological tumour type and grade are major determinants of spontaneous apoptosis in ovarian carcinomas; p53 alterations do not adversely but rather positively affect spontaneous apoptosis by increasing growth fraction. This, in turn, suggests p53-independency of spontaneous apoptosis in ovarian carcinomas.

A survey of the current management of varicose veins by members of the Vascular Surgical Society.

The assessment and treatment of varicose veins by members of the Vascular Surgical Society of Great Britain and Ireland has been assessed by postal questionnaire. The response rate was 65%, of which 77% were general surgeons with a vascular interest, 21% were vascular surgeons only and 2% were non-vascular. Approximately four new patients with varicose veins are seen per surgeon per week in clinics with a median waiting time to be seen of 12 weeks. A median of three varicose vein operations per surgeon per week are undertaken with 10-15% of surgery being performed for recurrent disease. The commonest indications for surgery are symptomatic (97%) and complicated (98%) varicose veins, although 55% of surgeons also perform surgery for cosmesis. 65% surgeons routinely use hand-held Doppler in the assessment of varicose veins; of the other methods available, Duplex scanning was used as the first line investigation by 83%. Although over 60% of surgeons use sclerotherapy surgery is the preferred option for primary treatment for varicose veins associated with long or short saphenous reflux. 62% surgeons use deep venous thrombosis prophylaxis in patients undergoing varicose veins surgery selectively, and 27% use it routinely.

Current management of acute leg ischaemia: results of an audit by the Vascular Surgical Society of Great Britain and Ireland.

BACKGROUND: Management of acute leg ischaemia has changed in recent years. This study aimed to elucidate current practice throughout the UK and Ireland. METHODS: Surgeons and audit departments were asked to return a questionnaire about every episode of acute leg ischaemia seen in the hospital between 1 January and 31 March 1996. RESULTS: A total of 539 episodes were reported in 474 patients (248 men) aged 19-96 (median 73) years. Common causes were thrombosis in situ (41 per cent), embolism (38 per cent) and graft or angioplasty occlusion (15 per cent). Vascular surgical advice was requested in 95 per cent of cases. Initial management was: immediate embolectomy in 21 per cent, anticoagulants in 13 per cent and no vascular intervention in 10 per cent. Arteriography was done in 56 per cent, followed by 186 endovascular and 165 surgical interventions. At 30 days, 70 per cent of limbs were definitely viable and 16 per cent had been amputated. The mortality rate was 22 per cent. Cases were reported by 86 of 182 hospitals contacted, but some referred no patients, and a supplementary audit of 54 cases (10 per cent size of the original sample) from non-contributing hospitals showed no important differences. CONCLUSION: Patients with acute leg ischaemia are generally treated by vascular specialists, with modern methods and acceptable results. This is being achieved despite insufficient vascular surgeons and radiologists for formal emergency rotas in most hospitals.

Prospective study of carotid endarterectomy and contralateral carotid occlusion.

A total of 700 patients who had carotid endarterectomy (CEA) in the UK and Ireland during a 6-month interval between March and August 1994 were studied prospectively. Some 108 patients (15.4 per cent) had a contralateral internal carotid artery occlusion. Previous reports have shown an associated stroke rate of about 10 per cent in these patients. This study assessed complications and outcome for patients undergoing CEA with contralateral internal carotid artery occlusion compared with those without. The indications for surgery were comparable between the two groups although the patients with occlusion had a slightly higher incidence of arrhythmia and stroke. Intraoperative shunts were used in a significantly higher proportion of those with occlusion (83.3 versus 64.7 per cent, P = 0.0001). The combined death and stroke rate for patients with occlusion was 5.6 per cent compared with 2.4 per cent for the remainder (P not significant). On the basis of the present data, CEA with a contralateral carotid artery occlusion carries only a slight increase in the rate of postoperative stroke and death. This increase was not statistically significant and is lower than that reported previously.

Efficacy of repeated pharmacotherapy in experimental acute poisonings with fluostigmine.

The effectiveness of repeated pharamacotherapy in acute poisonings with fluostigmine was studied in rats. It was demonstrated, that the second i.p. administration of obidoxime (40 mg/kg) was without effect. Repeated i.p. administration of atropine (10 mg/kg) shown marked efficacy only in animals treated previously with atropine (10 mg/kg) and diazepam (2.5 mg/kg). This procedure was without effect in animals treated just after intoxication with atropine (10 mg/kg) or atropine (10 mg/kg) and obidoxime (40 mg/kg). Subsequent administration of diazepam (2.5 mg/kg) or caramiphen (10 mg/kg) was without effect.

Inhibitory effects of beta-hydroxyethyl 2,4-dinitrophenyl disulphide on rat brain cholinesterase activity.

Hydrolysis of 1-6 mM AChJ by rat brain homogenates in TRIS-HCl buffer, pH 7-8, supplemented with NaCl and MgCl2, was found to be suppressed by a half by pretreatment with 0-3 mM beta-hydroxyethyl 2,4-dinitrophenyl disulphide (HEDD), a reagent for SH groups. This effect has been shown to be due to partial inhibition of a Phg-sensitive esterase which has been characterized as AChE on the ground of experiments with MeCh and BuCh. Similar inhibition of AChJ hydrolysis was also noted on pretreatment of brain homogenates with 0-05 M N-ethylmaleinimide (NEM). The inhibition degree has turned out to depend upon either AChJ concentration (0-1--4-8 mM) and duration of contact of the tissue with HEDD before addition of the substrate. The AChE activity revealed in 0-2 mM AChJ was found to be particularly sensitive to HEDD. Sensitivity of rat brain AChE to either HEDD and NEM may indicate for occurring of SH in some one its isozyme. No less, other mechanisms of the inhibition, particularly possibility of interaction of HEDD with a disulphide group must be also considered.