SPR imaging biosensor for determination of laminin-5 as a potential cancer marker in biological material.

A new method for the selective determination of laminin-5 concentration using a biosensor and surface plasmon resonance imaging (SPRI) technique is presented. A biosensor based on the specific interaction of laminin-5 with rabbit polyclonal antibody was constructed. The analytically useful dynamic response range of the biosensor is between 0.014 and 0.1 ng mL(-1). The detection limit is 4 pg mL(-1). The potential influence of interferences on the SPRI signal was investigated, and the high selectivity of the biosensor was confirmed. In order to demonstrate the potential application of the biosensor, laminin-5 concentration in blood plasma was determined. The results were compared with the laminin-5 concentration obtained by the commercial enzyme-linked immunosorbent assay (ELISA) kit. A comparison of results from healthy donors obtained by SPRI measurement and ELISA indicates that they are close and shows good agreement with the data reported in the literature. The plasma samples of bladder cancer patients gave higher concentration measured with specific biosensor than by ELISA assay. The study shows the clear difference in concentration of laminin-5 in healthy humans and patients with bladder cancer. Extensive clinical studies using the newly developed method can result in an increase in the use of laminin-5 as a potential cancer marker.

Carbohydrate-deficient transferrin depends on disease activity in rheumatoid arthritis and systemic sclerosis.

OBJECTIVES: Changes in the glycosylation of plasma proteins have been linked to the aetiology of the rheumatic diseases. The aim of this study was to determine and compare the levels of carbohydrate-deficient transferrin (CDT) in patients with rheumatoid arthritis (RA), systemic sclerosis (SSc), and systemic lupus erythematosus (SLE). METHOD: Studies were carried out in 29 female patients with RA, 27 with SSc, and 17 with SLE. CDT was assayed by the N Latex CDT immunonephelometric assay. RESULTS: The levels of %CDT in the sera of RA, SLE, and SSc patients were significantly higher than in controls while the absolute concentrations of CDT were unchanged. %CDT, CDT, and transferrin do not differ significantly between patients with rheumatic diseases. In RA and SSc patients, a positive correlation was observed between %CDT and C-reactive protein (CRP), as well as a positive correlation in RA patients between %CDT and 28-joint Disease Activity Score (DAS28). CONCLUSIONS: The changes in the serum %CDT concentration in patients with RA and SSc correlated with disease activity markers.

Relationship between CDT and disease activity in rheumatoid arthritis.

The aim of this study was to estimate the serum concentration of carbohydrate-deficient transferrin (CDT) in patients with rheumatoid arthritis (RA) and the relationship between the CDT level and disease activity in RA patients. Studies were carried out in 47 female patients with RA and 32 healthy women. Disease activity of RA was evaluated using the 28-joint count Disease Activity Score (DAS 28). Serum CDT was determined by particle-enhanced immunononephelometry using the N Latex CDT test. Patients with RA had significantly lower serum concentrations of CDT compared with controls. The correlation study showed the significant negative relationship between CDT and DAS 28 (r = - 0.483, p = 0.011). There were no correlations between serum CDT level and patient's age, disease duration, number of tender and swollen joints, and degree of disability evaluated by the Health Assessment Questionnaire. The level of CDT in patients with RA was significantly decreased and confirms the changes in transferrin glycosylation which are dependent on the disease activity. Therefore, measurement of CDT in the sera of patients with RA can be useful for the evaluation of disease activity in these patients.

Interleukin-6, soluble interleukin-2 receptor and soluble interleukin-6 receptor in the sera of patients with different histological patterns of rheumatoid synovitis.

OBJECTIVE: The present study was conducted to investigate whether the serum levels of interleukin 6 (IL-6), soluble IL-2 receptor (sIL-2R) and sIL-6R are associated with the morphological appearance of rheumatoid arthritis (RA). METHODS: Using the ELISA technique we measured the IL-6, sIL-2R and sIL-6R concentrations in the serum of 34 patients with RA and 28 patients with osteoarthritis (OA). Histological analysis of synovial samples distinguished 2 types of rheumatoid synovitis. Twenty-one RA specimens presented diffuse infiltrates of mononuclear cells without any specific microanatomical organization. In remaining 13 samples the formation of lymphocytic follicles with germinal center-like structures was found. RESULTS: Serum levels of IL-6, sIL-2R and sIL-6R were elevated in patients with RA compared to the OA control group (p < 0.001, p < 0.001 and p < 0.05 respectively). Concentrations of IL-6 and sIL-2R were highest in the serum of RA patients with follicular synovitis in comparison to patients with diffuse synovitis (p < 0.001 and p < 0.01 respectively) and could distinguish RA patients with these two histological variants of the disease. Serum levels of IL-6 and sIL-2R correlated with markers of disease activity such as ESR and CRP levels. In addition, the clinical data suggest a more severe disease among RA patients with follicular synovitis. CONCLUSION: Distinct histological types of rheumatoid synovitis associated with unique serum concentrations of IL-6 and sIL-2R reflect levels of disease activity and confirm the concept of RA heterogeneity.

Malignant neoplasms of the pancreas. A study based on autopsy data from 1953 to 1982 in Bialystok, Poland. I. Frequency, age and sex distribution.

Among 18 751 autopsies in persons over 20 years of age 195 primary malignant neoplasms of pancreas (MNP) were stated (117 in males, 78 in females; the ratio being 1.5 : 1). In the period of the investigation (1953--1982) MNP comprised 1.0% of all autopsies, 4.1% of all malignant neoplasms (MN), and 11.0% of MN of alimentary system (MNAS). The frequency of MNP in all autopsies was from 0.5% during the years 1953--1957 to 1.2% in 1978--1982, in all MN from 2.7% to 4.5%, and in all MNAS from 5.7% to 12.7%, respectively. MNP were observed most frequently at the age of 61--70. The mean age was 63.6 and was higher in females than in males (66.3 and 61.8, respectively), and increased in the period under study from 54.6 to 65.6 (in males from 50.8 to 64.8, in females from 59.3 to 66.8). The significant increase in the incidence of MNP in the population of patients who died in all hospitals in Bialystok (the north-eastern Poland) between 1953--1982 and were autopsied in our Department correlates with data indicating the rise in the incidence of MNP in many countries of the world.

Malignant neoplasms of the pancreas. A study based on autopsy data from 1953 to 1982 in Bialystok, Poland. II. A survey of 195 cases.

Among 195 malignant neoplasms of the pancreas (MNP) diagnosed at autopsy in 1953--1982 the exocrine carcinomas comprised 85.6% and inmular carcinomas 13.8% of cases. MNP were localized most frequently in the head of the pancreas (54.4%), then in the whole pancreas in 14.9%, in the head and the body or in the body and the tail in 20.5%, in the body or in the tail in 10.3% of MNP. There were no infiltrations and no metastases in other organs only in 16 patients (8.2% of MNP). The biliary tracts and duodenum were the most frequent sites of secondary infiltrations. Metastases were most frequent in the liver, then in the liver hilus and mesentery lymph nodes. The concordance of clinical and post mortem diagnosis of MNP was stated in 40.5% cases.

The effect of repeated BCG vaccine administration associated with surgery upon the growth of lung metastases in rats with transplantable Morris 5123 hepatoma.

Eighty four male Buffalo rats bearing transplantable Morris 5123 hepatoma were used for the experiment. Half of the animals were given into the tumor BCG vaccine (final dose 2.2 X 10(6) living mycobacteria), the other 42 rats after BCG underwent surgical removal of the hind leg together with the tumor. Though the immunotherapy reduced the size of lung metastases of the tumor it did not, however diminish the survival rate of the animals. Multiple BCG doses associated with the surgical procedure prevented dissemination of the tumor cells and prolonged the survival of rats up to the 100th day of the follow-up.

Von Willebrand factor antigen in assessment of vasculitis in patients with connective tissue diseases.

Von Willebrand factor antigen (vWF:Ag) is synthesized and secreted by endothelial cells. In the present study we tried to assess the relationship between plasma level of vWF:Ag and vascular damage in patients with vasculitis. The study was carried out on 59 patients with connective tissue diseases. Vasculitis was diagnosed by biopsies of the skin. The patients with vasculitis had a significantly elevated level of vWF:Ag; however, no significant correlation between the amount of plasma vWF:Ag and the degree of vasculitis was found. The obtained results show that the plasma level of vWF:Ag may reflect the presence of vascular, especially endothelial, damage in patients with connective tissue diseases.

AgNORs in duct epithelial lesions in chronic pancreatitis and in pancreas cancer cells.

BACKGROUND/AIMS: Argyrophilic nucleolar organizer regions (AgNORs) reflect the proliferative activity of cells. Since the majority of pancreatic cancers are ductal carcinomas, the aim of the study was to determine the AgNORs expression of potential pre-neoplastic ductal epithelial lesions in advanced chronic pancreatitis compared with pancreatic cancer cells. METHODOLOGY: Histological preparations obtained from 24 patients with chronic pancreatitis and 16 patients with pancreatic cancer were used to estimate the number of AgNORs per nucleus. Four types of AgNORs were distinguished and histograms with cell percentage of each type were performed for all forms of epithelial anomalies. RESULTS: In simple hyperplasia, squamous and mucous metaplasia the number of AgNORs ranged from 1.92 to 2.23; type I was predominant. In papillary hyperplasia, dysplasia and in situ carcinoma the number ranged from 2.98 to 3.34, with a predominance of type II-IV. In invasive carcinoma the number was 4.29 and 74% of cells were of type II-IV. CONCLUSIONS: Both counts of AgNORs and the percentage of type II-IV cells showed a gradual increase from simple hyperplasia through papillary hyperplasia and dysplasia to invasive carcinoma which in this respect differs significantly from all forms of the epithelial anomalies examined.

Epithelial anomalies in chronic pancreatitis as a risk factor of pancreatic cancer.

BACKGROUND/AIMS: The relationship between chronic pancreatitis and the development of pancreatic cancer is still a matter of dispute. Our aim was to determine the frequency of hyperplastic, metaplastic and dysplastic epithelial anomalies in the course of chronic pancreatitis and the potential steps in their development to malignancy. METHODOLOGY: The study was based on biopsy material of 70 patients with clinically diagnosed advanced chronic pancreatitis, who underwent partial or total pancreatectomy, as well as other operations. The patients were assigned to 2 groups: Group I (n = 41) with calcifying chronic pancreatitis; Group II (n = 29) with other forms of the disease. Histological sections were stained with hematoxylin-eosin, Mallory-azan, Gomori's silver method, and glycosaminoglycans (PAS and Alcian blue staining). Special interest was focused on the type and incidence of epithelial ductal and acinar cell anomalies, and on the degree of parenchymal scarring. RESULTS: Hyperplasia of the ductal epithelium was present in 31.4%, focal squamous metaplasia in 21.4%, mucous metaplasia in 11.1%, cellular dysplasia in 8.6%, dysplastic acinar cell nodules in 21.4%, and "tubular complexes" in 30.0% of all cases. The differences in the frequency of these changes, except for ductal epithelial hyperplasia, were not statistically significant in two comparable groups. Advanced pancreatic fibrosis was associated with epithelial anomalies in 65.7% of all cases. CONCLUSIONS: From the morphological point of view, the adequate prerequisites for the consideration of advanced forms of chronic pancreatitis, independent of type, as a risk factor of pancreatic cancer exist, necessitating the surgical removal of pathological lesions.

Morphometric and ultrastructural studies of the sciatic nerve regeneration in rats intoxicated with ethanol.

The aim of the study was to examine the process of sciatic nerve regeneration and changes in the dorsal root ganglia (from which sensory fibres of the sciatic nerve extend) in animals intoxicated with ethanol. The experiment used 20 rats, divided into two groups: control and treated. The treated animals were intragastrically given 2g/kg b.w. of ethanol in 25% aqueous solution. In both groups the right sciatic nerve was transected and then sutured. After 5 months the animals were anaesthetized. The left and the right spinal dorsal ganglia-L5 and sections from the non-operated and operated sciatic nerves were collected for analysis. Ultrastructural examinations and morphometric measurements were conducted. It was found that ethanol administrated to rats inhibited regeneration of the transected and then sutured sciatic nerve, impairing the growth of axons in the transected nerve and destroying the regenerating sensory ganglion cells. The mechanism of the changes described may be associated with axonal transport disorders or with the suppressed production of biologically active substances, which affect nerve regeneration.

Morphometric and histological examinations of dog kidneys after embolization of the renal artery with the cyanoacrylic glue "Chirurcoll-Polfa".

The renal artery in 20 mongrel dogs was embolized with cyanoacrylic glue "Chirurcoll-Polfa", introduced through a catheter. The extent of the resulting renal infarction of the kidneys was evaluated morphometrically and was found to be 71.35 to 91.43% of the volume of the embolized kidneys (mean 77.61%). Histological examinations revealed no resorption of the glue after 3 months. In the embolized vessels, after 7 days tumefaction and degeneration of the reticulin, elastic, and collagen fibres were observed, after 24 days inflammatory changes, and after 2 and 3 months respectively, secondary collagenization and hyalinization. Inflammatory changes were also found in the perivascular tissue. In the sections of the kidneys not involved in the infarction a progressive atrophy of the nephrons and fibrosis of the parenchyma were observed 2 and 3 months after embolization.

Histological and histochemical changes in the dog kidney after renal artery embolization with Spongostan.

Histological and histochemical examinations of canine kidneys 24 hours, 7, 14 and 24 days after embolization of the renal artery with Spongostan showed that during 24 days the resorption and organization of the infarcts in the embolized kidneys and the resorption of Spongostan in the arteries were not yet completed. The intensified reactions to phosphatases, dehydrogenases, and non-specific esterase indicate a prolonged activation of cells playing a role in the resorption and organization of infarcts which may be of significance in the stimulation of immune responses. There were no inflammatory reactions in the embolized arteries.

Morphometric investigations of dog kidney infarcts after embolization of the renal artery with Spongostan.

A macroscopic morphometric method for evaluating the volume of infarcts in the kidneys of dogs 24 hours, 7, 14, and 24 days after renal artery embolization by means of Spongostan is described. The whole kidney was not infarcted, the volume of the infarcts being 60.15-81.87% (mean 71.1%) of the volume of the embolized kidney. In order to evaluate the distant morphological changes in the parts of the embolized kidney not involved in the infarcts, it would be necessary to carry out microscopic examinations and to have a longer observation period.

Effect of clamping of the renal vein in dogs on certain biochemical and histopathological changes.

Experimental investigations were carried out on dogs in which the renal vein was clamped for a period of 15 min, 30, and 60 min consecutively. It has been found that clamping of the renal vein for 15 min results in tissue anoxia and degeneration of the epithelium of the renal tubules. Clamping for 60 min causes cellular acidosis and necrosis of the tubules. It can therefore be said that clamping of the renal vein for even a short time is highly dangerous.

The effect of internal iliac artery embolization of the prostatic gland in the dog.

Investigations were carried out on dogs in order to study the changes in the prostatic gland after embolization of the internal iliac arteries. The experiments showed that slight fine-celled infiltrations occurred in the interstitial tissue as a result of ischaemia of that organ. The authors are of the opinion that embolization of the internal iliac arteries may be used in clinical practice as a method of treating haemorrhages from the prostatic gland.

The use of tissue adhesive "Chirurcoll-Polfa" in surgical fixation of the kidney.

Experimental and clinical investigations were carried out on the application of tissue adhesive "Chirurcoll-Polfa" in surgical fixation of the kidney. It was found that the adhesive gives firm tissue adhesion without causing inflammatory reaction, and can be safely used in operations for fixation of the kidneys.

[Expression of p53 protein and proliferative activity in multiform glioblastoma]. / Ekspresja bialka p53 i aktywnosc proliferacyjna w niedojrzalych glejakach wielopostaciowych.

The aim of the study of 41 multiform glioblastomas was the analysis of p53-protein immunoreactivity in neoplastic cells and evaluation of relationship of this biologic marker to tumour proliferation activity. Positive p53 expression was observed in 24 (58.5%) tumours, the negative one in 17 tumours (41.5%). Proliferation indexes of PCNA, anti-Ki6 and AgNORs showed high values in the multiform glioblastoma p53 positive group, but without statistical differences in comparison with the group of p53-negative glioblastomas. Significant differences were observed in survival time of patients with p53 positive tumours in comparison with p53-negative ones. In 15 patients with p53-positive multiform glioblastomas survival time was less than 6 months (62.5%) on the contrary with only 4 patients with similar survival time in p53-negative glioblastoma group (23.5%). Our results suggest that p53 expression in multiform glioblastoma cells, generally considered as the indirect index of p53 suppressor gene, reflects aggressive stadium of neoplastic disease and significantly worsens the prognosis.

[Proliferative activity of glial neoplasms of the brain]. / Badania aktywnosci proliferacyjnej glejowych nowotworów mózgu.

The aim of the study including 89 brain gliomas was to determine their proliferative activity assayed with immunohistochemical methods (PCNA and Ki-67) and with the method of AgNORs, as well as to evaluate the correlation between the proliferative activity and features of histological malignancy. The study reveals that the estimation of PCNA, Ki-67 and AgNORs are effective methods for the determination of the proliferative activity of brain gliomas. Statistically significant differences were noted in the proliferative PCNA, Ki-67 and AgNORs between groups of gliomas with lower and higher malignancy, which indicated a distinct correlation between histological malignancy of the tumours and their proliferative activity. High values of PCNA and Ki-67 (> 40%) and AgNORs (> 15) were found to considerably deteriorate prognosis in brain gliomas.

[Primary liver cell carcinoma among the population of Bialystok in 1975-1988 in relation to hepatitis B virus infection]. / Pierwotny rak watrobowokomórkowy w populacji Bialegostoku w latach 1975-1988, w relacji z zakazeniem wirusem hepatitis B.

In presented study, retrospective epidemiological analysis was performed in 84 patients dead due primary hepatocellular carcinoma, selected from among 17,973 persons dead in Bialystok hospitals from 1975 till 1988. In the part from among these patients was estimated prevalence of serological markers of Hepatitis B virus infection, as a potential oncogenic factor. Analysis of this carcinoma diagnosis frequency in particular years showed rising tendency: from 1975 till 1979 number of PHC diagnosis was from 0 to 5 a year, whereas in 1988 it was 13. Percentage of necropsy diagnosed PHC was similar and increased to over 1% of all necropsies in 1988. Mean age of patients dead due PHC was 65, and 71.4% were men. Serological markers of HBV infection were observed in 50% and HBsAg in 34.6% from among PHC-dead patients. These percentage values were significantly higher, than observed in control group of Bialystok population, which indicate possibility of association between HBV infection and PHC development.