Cathepsin D-related disease-free interval in pT1 primary breast carcinomas: a pilot study.

In order to address the heterogeneity of the pT1 breast cancer stages, we have been examining the natural and the clinical course of disease in relation to cathepsin D expression, as a molecular marker for the tumor progression that leads to metastasis. The original aim of our pilot study was to determine whether it was possible to distinguish high-risk from low-risk patients, on the basis of nonestrogen- vs. estrogen-regulated cathepsin D expression. Our results showed that estrogen-regulated cathepsin D expression could be useful as surrogate marker of node-positive status. Further, during the natural course of disease, none of 7 pT1N0 patients with tumors bearing nonestrogen-regulated cathepsin D expression developed metastasis. During the clinical course of disease, nonestrogen-regulated cathepsin D expression defined low-risk while estrogen-regulated cathepsin D expression defined high-risk pT1N+ subgroup of patients. Although there is no consensus with respect to metastasis-related prognostic value of cathepsin D expression, our pilot study implies its prognostic value in pT1 breast cancer patients and supports the hypothesis that cathepsin D may promote metastasis in this early stage of disease.

[Radionuclide detection of Helicobacter pylori infection].

The aim is the assessment of the HP infection in stomach using breath test and comparison to other diagnostic methods, as well as following up the effect of therapy. In 83 patients with digestive discomfort rapid urease test, histology and breath test were performed, while in 25 patients with proven HP infection the effect of therapy was followed up using breath test and clinical findings. For rapid urease test and histology, samples were taken from antral mucosa. Breath test was performed after per oral administration of the capsule of 14C- urea (37 kBq) (Izotop, Hungary and Laboratory for radioactive isotopes, Vinca) which, in the presence of Helicobacter pylori breaks up to 14CO2 and NH3. Radioactivity was measured by beta counter in the exhaled air fasting and 30 minutes after ingestion of the capsule. According to our results, the rise of activity over 100% was considered positive. From 83 patients, 58 were breath test was positive, 24 negative and one equivocal. Fast urease test was in 54 positive, in 29 negative while histology was in 57 postitive and 26 negative. Findings of the breath and urease tests were in accordance in 93% patients while breath test and histology in 98% patients. During follow up of the therapeutic effects, breath test and clinical findings were in accordance in 98% patients. Breath test can be useful in diagnosis but is a method of choice in following up the patients after therapy for H. pylori infection, because it is non-invasive, fast and precise.

Radioimmunodetection of colorectal carcinoma.

BACKGROUND/AIMS: The aim of the study is detection of the recurrences and metastases of colorectal carcinomas using 111In-labeled antibodies B72.3. METHODOLOGY: Fourteen patients underwent planar immunoscintigraphy and/or tomoscintigraphy. RESULTS: With tomography in comparison to planar scintigraphy, we can access better distinction of tumor and estimation of its size. Other imaging methods (computed tomography, ultrasonography) have an advantage in detection of liver metastases, while immunoscintigraphy is more specific for the assessment of malignant abdominal tumors and extrahepatic metastases. CONCLUSIONS: The first results point out that Oncoscint CR-103 can be useful in diagnosis of recurrences and metastases of colorectal carcinoma, viability assessment after radiotherapy and in the choice of the adequate surgical treatment in dependence of the spread of the disease.

[Indium 111-labeled antibodies in the detection of colorectal carcinoma]. / (111)In obelezena antitela u otkrivanju kolorektalnog karcinoma.

The aim of the study is detection of the recurrences and metastases of colorectal carcinomas using (111)In labelled antibodies B72.3 in 14 patients. With tomography, we can access better distinction of tumour in comparison to other structures and estimation of its size. Other imaging methods (CT, US) have advantage in detection of liver metastases, while immunoscintigraphy is more specific for the assessment of malignant abdominal tumours and extrahepatic metastases. The first results point out that Oncoscint CR-103 can be useful in diagnosis of recurrences and metastases of colorectal carcinoma, viability assessment after radiotherapy and in the choice of the adequate surgical treatment in dependence of the spread of the disease.

Local recurrence and five year survival after abdominoperineal resection of the rectum due to rectal carcinoma.

In the period 01.01.1991-12.31.1996, 523 operations due to rectal carcinoma were performed on the First Surgical Clinic, the Third Department for Colorectal Surgery. Most common localization of tumor was in the distal third of the rectum 65.2%. In the middle third, there were 28.9% and in the upper, intraperitoneal third 5.9%. We performed 286 low anterior stapled resections, 93 anterior resections with hand-sewn anastomosis and 144 Abdominoperineal excisions of rectum (Miles procedure). Pathohistological examination revealed adenocarcinoma in all cases. In this study we analyzed local recurrence and five-year survival after long-term follow-up in the group where Miles procedure was carried out as a potentially curative procedure (except 4.9% cased with Dukes D stage). There were 74.3% males and 23.7% females median age 59.2 years. According to Dukes classification there were 4.9% in stage A, 47.2% in stage B, 43.1% stage C, and 4.9% stage D. There were 4(2.7%) postoperative deaths. Recurrence of the disease was registered in 44 (30.5%) patients. Local recurrence alone was found in 14 (9.7%) patients, while distant spread was registered in 30 (20.8%) patients. At present, the median follow-up is at 72.9 months. Analysis by the Kaplan-Meier's test shows cumulative survival of 61%, and disease free survival of 63.4% at 60 months of the follow-up. Dukes C is associated with a very poor prognosis; survival after 60 months of follow up shows cumulative Survival of 0.35 while Dukes B has far better prognosis (0.86). Analysis of disease free survival by Dukes stage shows that Dukes C has the worst prognosis (disease free survival 0.36 after 60 months), while stage B has much better prognosis (0.84). Local recurrence analysis by the Kaplan-Meier's test shows disease free survival of 84.9% at 60 months of follow-up. Analysis of local recurrence by Dukes stage shows 1.00% disease free survival for cases in stage A, 0.94 for Dukes B and 0.66 for Dukes C, while overall comparison between groups regarding local recurrence using the Wilcoxon (Gehan) statistic shows statistically significant difference (p-0.005). There is no statistical difference between Dukes A and Dukes B cases in distribution of local recurrence.

PS2 protein in breast carcinomas: cut-off value of estrogen-regulated expression.

This study includes 152 patients with histologically confirmed breast carcinoma. Steroid hormone receptors (SR), estrogen (ER) and progesteron (PR) receptors, and pS2 protein were assayed on the same cytosolic extract in accordance with the recommendation of EORTC. Our results showed menopausal- and histologic grade-related expression of pS2 protein. Unfavorable carcinoma subgroups, in relation to expression of pS2 protein were defined: postmenopausal carcinomas with histologic grade II, and pre-, as well as postmenopausal carcinomas with histologic grade III. There were overlappings of individual pS2 protein values between favorable and unfavorable carcinoma subgroups in relation to the expression of pS2 protein. Otherwise, no overlapping of pS2 protein values was obtained between ER-positive and ER-negative carcinomas within defined unfavorable menopausal - and histologic grade-related expression of pS2 protein subgroups. The highest pS2 protein level observed in ER-negative unfavorable subgroups (15 ng/mg) was considered as the cut-off value which defined estrogen-regulated expression of pS2 protein.

14C-urea breath test in the detection of Helicobacter pylori infection.

Helicobacter pylori infection is supposed to be one of the major causes of digestive and other diseases. Among a lot of invasive and non-invasive methods for its detection, none is ideal. The aim is an assessment of the Helicobacter pylori infection in the stomach using breath test and comparison to other diagnostic methods, as well as following up the effects of therapy. In 17 patients with digestive discomfort, breath test, rapid urease test and histology were performed, while in 47 patients with proven HP infection the effect of therapy was followed up using breath test and clinical findings. Breath test was performed after per oral administration of the capsule of (14)C urea (37 kBq). Findings of the breath and urease tests were in accordance in 14/17 patients (83%) while breath test and histology in 16/17 patients (94%). During follow-up of the therapeutic effects, breath test and clinical findings were in accordance in 43/47 patients (98%). Breath test can be useful in diagnosis but is a method of choice in following up the patients after therapy for H. pylori infection, because it is non-invasive, fast and precise.

Long-term results after low anterior stapled anastomosis.

In the period 01.01.1991-12.31.1996, 286 low anterior stapled resections of the rectum due to rectal carcinoma were performed at the First Surgical Clinic, the Third Department for Colorectal Surgery, Belgrade. There were 57% males and 43% females, median age 59.6 years. The most common localization of tumor was in the distal third of the rectum 181 (63%). In the middle third, there were 89 (31%) and in the upper, intraperitoneal third 16 (6%). Histopathological examination revealed adenocarcinoma in all cases. All operative specimens were examined by one pathologist and classified according to the Gunderson-Sosin modification of Dukes classification. There were 14 (4.9%) in stage A, 167 (58.4%) in stage B (B1,B2,B3), 89 (31.1%) stage C (C1,C2,C3) and 16 (5.6%) stage D. According to Broders classification, there were 129 (45%) well differentiated, 142 (50%) moderately and 15 (5%) poorly differentiated tumors. Anastomotic dehiscence was found in 17 patients (5.95%), mostly conservatively treated, except in 6 cases where spreading peritonitis developed requiring operative treatment. There were 9 (3.1%) postoperative deaths, a half of them with specifically operation related mortality. Recurrence of the disease was registered in 47 (18%) patients, out of 260 who were regularly followed up (26 were lost). Local recurrence alone was found in 21 (8.0%) patients, while distant spread was registered in 22 (8.46%) patients. Local and distant spread was found in 4 (1.5%) cases. At present, the median follow-up is at 54 months. Analysis by the Kaplan-Meier's test shows cumulative survival of 66% at 71 months of the follow-up. Seventy four percent of patients with curative operations exhibit no signs of recurrence at 5 years.

Cathepsin D: association between TN-stage and steroid receptor status of breast carcinoma.

PURPOSE: To assess whether the expression of estrogen-induced protease, cathepsin D, might facilitate biological subgrouping of patients with breast carcinomas, and accordingly, its potential applicability in clinical oncology. PATIENTS AND METHODS: This study includes 70 patients with histologically confirmed breast carcinoma. Pathological findings were classified according to tumor size (T) and the presence or absence of metastases in regional lymph nodes (N). Steroid hormone receptor (SR) density as well as cathepsin D concentrations were assayed in the cytosol of breast carcinomas in accordance with the recommendation of the EORTC. RESULTS AND DISCUSSION: Statistically significant direct correlations were observed between cathepsin D expression and axillary node status as well as SR status. However, it is important to point out that in spite of these statistically significant findings, there were no biologically significant associations due to a wide range of individual cathepsin D values. Baseline levels of cathepsin D expression were found in patients with SR-negative status and node-negative tumors as well as in patients with SR-negative status and tumors of

pS2 in breast carcinoma: association with steroid hormone receptor status.

AIMS AND BACKGROUND: Knowledge of the steroid hormone receptors has proved to be of significant value in breast cancer. In the present study the possible importance of estrogen-regulated pS2 protein was investigated. Our direct purpose was to answer the question whether the expression of pS2 may be a marker of functional heterogeneity with respect to the steroid hormone receptor status. METHODS AND STUDY DESIGN: The study included 152 patients with primary, operable, histologically confirmed breast carcinomas. Histology specimens were reviewed and classified according to type, nodal status, tumor size and grade. Steroid hormone receptors were assayed by biochemical methods according to the procedures recommended by the EORTC. pS2 protein measurement was performed in breast carcinoma cytosols using an immunoradiometric assay. The results were analyzed by non-parametric statistical methods. RESULTS: A statistically significant inverse correlation between pS2 protein expression and histological tumor grade was found. The expression of pS2 protein was confirmed to be correlated with steroid hormone receptor status. However, it is important to point out that in spite of these statistically significant findings there were no significant biological associations due to overlapping individual pS2 protein values. The baseline level of expression of pS2 protein was obtained in histological grade III carcinomas with a negative steroid hormone receptor status. It was shown that the distribution of carcinomas according to the baseline level of pS2 protein expression was heterogeneous among estrogen receptor-positive carcinomas, and strikingly homogeneous among estrogen and progesterone-negative carcinoma. CONCLUSION: Our study suggested that PR and pS2 protein may identify distinct subsets of estrogen receptor-positive breast carcinomas.

[The effect of hemodialysis and continuous ambulatory peritoneal dialysis on renal anemia]. / Utcaj hemodijalize i kontinuisane ambulatorne peritoneumske dijalize na anemiju bubreznih bolesnika.

Anaemia is an almost invariable sign of chronic renal failure [1]. Although many factors have been implicated as causes of this anaemia, it seems probable that deficiency of erythropoietin is the main cause for most patients [2]. Institution of chronic dialysis can improve anaemia in end-stage kidney disease, continuous ambulatory peritoneal dialysis being reported as more successful [3]. The aim of this study was to investigate the influence of haemodialysis and continuous ambulatory peritoneal dialysis on anaemia during the first six months of treatment. We examined 21 persons (14 males and 7 females, aged from 18 to 78 years) on haemodialysis treatment and 13 persons (6 males and 7 females aged from 22 to 64 years) on continuous ambulatory peritoneal dialysis (Table 1). Standard procedures were used for measuring biochemical parameters. Urea and creatinine levels were high, almost incompatible with life, in all tested persons before dialysis treatment. During the first three months of both dialysis techniques urea and creatinine were significantly (p < 0.01) corrected, but remained above the normal ranges (Table 2). Patients on continuous ambulatory peritoneal dialysis have shown significantly (p < 0.01) lower urea and creatinine values compared to patients on haemodialysis (Graph 1). These data suggest better preservation of renal function and better control of the internal environment during continuous ambulatory peritoneal dialysis [6]. All tested patients were severely anaemic before the beginning of dialysis. During the first six months of haemodialysis erythrocyte count, haematocrit and haemoglobin levels were unchanged (Table 3). Transfusions and hepatitis episodes only temporary improved anaemia. Patients on continuous ambulatory peritoneal dialysis exhibited significant correction of anaemia already during the first three months of treatment (Graph 2). Though less significantly, haemoglobin values continued to rise even during the next three months. The reached haemoglobin levels were lower than normal, but significantly higher than values in patients on haemodialysis (p < 0.01), suggesting better control of anaemia during continuous ambulatory peritoneal dialysis. Transfusion requirement was irrelevant, and hepatitis was not noticed, so they cannot be held responsible for the improvement of anaemia. Greater iron consumption, illustrated by higher transferrin saturation, also confirmed increased erythopoitesis in patients undergoing continuous ambulatory peritoneal dialysis. They also had lower blood iron level than those on haemodialysis (who had) numerous blood transfusions. The improvement of anaemia during continuous ambulatory peritoneal dialysis may be the result of reduction in plasma volume [7] as well as an increase in red cell mass and a better clearance of middle molecules in comparison to patients on haemodialysis. The main cause is higher erythropoietin level [8]. All tested patients had low folic acid level. Patients who corrected anaemia showed fall in folat level. This was statistically remarkable during the first three months of continuous ambulatory peritoneal dialysis-from 3.64 ng/ml to 2.09 ng/ml. All these data suggest that both dialysis modalities are effective in the control of protein waste products level, but continuous ambulatory peritoneal dialysis has better influence on the improvement of anaemia that haemodialysis. This can be attributed to better removal of uremic toxins, improved protein metabolism, lower parathyroid hormone level and higher erythropoietin value due to peritoneal macrophage production.

Endothelin-1 in acute renal failure.

To study the pathological significance of circulating endothelin (ET) in ARF, we measured plasma ET in seven children (mean age 8.8 +/- 4.4 years) with ARF in the most severe phase and 3.7 +/- 3.5 months later in the recovery period. Twenty-seven healthy children were included in the study as controls. Plasma ET level was measured by highly sensitive and specific radioimmunoassay for ET-1 and ET-2 (ET-1/2, Biomedica, Vienna). Plasma ET was significantly higher in the most severe phase of ARF (4.75 +/- 4.08 fM/ml) than in the recovery period (0.78 +/- 0.24 fM/ml; p < 0.01), but comparing to plasma ET in the healthy children, the difference was only of borderline statistical significance (Pf, 0.0573). Since plasma concentrations of creatinine did not correlate with plasma ET in patients, either in acute or in the recovery phase of disease, we concluded that decreased GFR is not the main factor determining an increased ET in ARF. We suggest that elevated plasma ET in ARF may be secondary to vascular endothelial dysfunction and speculate that enhancement synthesis of endothelial relaxing factor (EDRF) inhibits ET synthesis during the recovery period. We did not find any relationship between plasma ET and blood pressure (BP) in patients with ARF, so we conclude that circulating ET is not the main factor determining BP in ARF.

[The effect of hemodialysis on endothelin-1 plasma levels in children with terminal renal insufficiency]. / Uticaj hemodijalize na nivo endotelina-jedan u plazmi dece s terminalnom insuficijencijom bubrega.

We analysed the effect of hemodialysis (HD) on endothelin (ET) plasma level in children with terminal renal failure. Twenty-five patients (pts) and 27 healthy children as controls were included in the study. There was no difference in ET plasma levels in pts before HD (1.97 +/- 1.45) and controls (2.08 +/- 1.47), but ET increased in plasma of pts after HD (4.10 +/- 3.66). To estimate the relationship of volume depletion or depuration on ET plasma level changes in pts during HD, ET was measured at three time points of HD (before ultrafiltration-UF, after UF, and after HD without fluid removal) in 10 pts. ET level was not significantly changed after UF (1.93 +/- 2.25 vs 1.71 +/- 1.50; ns), but was significantly increased after depuration (4.46 +/- 3.56; p < 0.05). There was no correlation between ET and blood pressure in controls and pts in either period of testing, neither with plasma renin activity, left ventricular mass index, and body weight changes during HD.

[The role of endothelin in acute renal failure]. / Znacaj endotelina u akutnoj insuficijenciji bubrega.

During the study of the pathological significance of circulating endothelin (ET) in acute renal failure (ARF) we measured ET in 7 children (mean age 8.8+4.4 years) with ARF in the most severe phase, and 3.7+3.5 months later in the recovery period. Twenty-seven healthy children (mean age 11.1+/- 6.3 years) were included in the study as controls. Plasma ET level was measured by highly sensitive and specific radioimmunoassay for ET-1 and ET-2 (ET 1/2, Biomedica, Vienna). Plasma ET was significantly higher in the most severe phase of ARF than in the recovery period, but comparing to plasma ET in the healthy children, the difference was only on borderline statistical significance. Since plasma concentrations of creatinine did not correlate with plasma ET in patients, either in acute or in the recovery phase of disease, we concluded that decreased glomerular filtration (GFR) was not the main factor determining the increased ET in ARF. We suggest that elevated plasma ET in ARF may be secondary to vascular endothelial dysfunction and speculate that engancement synthesis of endothelial relaxing factor (EDRF) inhibits ET synthesis during the recovery period. The pathogenetic role of circulating ET in ARF cannot be determined from the present study.

[Significance of endothelin in arterial hypertension in children]. / Znacaj endotelina u arterijskoj hipertenziji kod dece.

UNLABELLED: Concentration of plasma endothelin and the activity of Na(+)-K(+)-ATP-ase were determined in 3 different groups of children: the 1st group comprised 13 children with essential hypertension, aged 12.9+/-4.8; the 2nd group concerned 16 children with renal hypertension, but with preserved global renal function, aged 13.7+/-2.8, and the 3rd group consisted of 27 healthy children, aged 11.6+/-6.3 years. Plasma endothelin concentrations were measured by radioimmunological method, using a set manufactured by "Biomedica". The activity of Na(+)-K(+)-ATPase was determined in erythrocyte haemolysate by measuring the quantity of released inorganic phosphate in samples with and without ouabaine. Concentration of plasma endothelin was not significantly different between the children with arterial hypertension and healthy children, and there was no significant correlation between endothelin concentration and blood pressure in either of the 3 groups of children. The activity of Na(+)-K(+)-ATP-ase was significantly decreased only in the 1st group of children. There was no evidence of correlation between the Na(+)-K(+)-ATPase activity and blood pressure or plasma endothelin either in healthy, or in hypertensive children. CONCLUSION: Endothelin in blood circulation has no significance in regulating blood pressure in healthy children. It also has no direct role on the development of essential and renal hypertension in children. The activity of Na(+)-K(+)-ATP-ase is decreased in children with essential hypertension, but it seems that it has no direct impact on hypertension. Endothelin in circulation and Na(+)-K(+)-ATP-ase activity are not interdependant.

The clinical use of 131I-meta-iodo-benzylguanidine (MIBG) for the diagnosis of neuroblastoma.

Whole-body scintigraphy with 131I-meta-iodo-benzylguanidine 131I-MIBG) was performed in 41 patients with neuroblastoma. In patients with clinical remission no pathological concentration of 131I-MIBG was found. In 30 patients with residual, recurrent or metastatic disease neuroblastoma was correctly localized by 131I-MIBG scintigraphy. It is concluded that 131I-MIBG whole-body scintigraphy is useful in the diagnosis and follow-up of neuroblastoma.

Elevated serum gastrin levels in patients with gastric cancer.

Elevated plasma gastrin levels have been found in patients with colorectal cancer. We measured fasting serum gastrin levels in control subjects (n = 12), patients with gastric cancer (n = 43), and patients with carcinoma of the esophagus (n = 55). Serum gastrin levels were significantly higher in patients with gastric cancer compared to normal controls (P less than 0.005) and those with esophageal cancer (P less than 0.05). This information may add to our understanding of the pathogenesis of gastric cancer.