Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer.

BACKGROUND: Gap junctions are intercellular channels composed of connexins, which mediate the direct passage of small molecules between neighbouring cells. They are involved in regulation of cell cycle, cell signalling, and differentiation, and probably invasion and metastasis. The role of connexins in the metastatic process is controversial, because some studies indicate that connexin expression is inversely correlated with metastatic capacity. In contrast, others demonstrate that connexins may be involved in metastasis. In addition, connexin status in breast cancer metastasis has not been widely studied. METHODS: We evaluated by immunohistochemistry the expression of connexin 26 (Cx26) and connexin 43 (Cx43) in primary breast tumours (PTs) and matched paired metastases to lymph nodes (MLNs). RESULTS: In PTs, we observed predominantly cytoplasmic localisation of evaluated connexins, indicating alterations in connexin expression in breast cancer cells. We demonstrated that expression of Cx26 and Cx43 was increased in MLNs compared with PTs (p<0.00001 and p<0.001, for CX26 and Cx43, respectively). In addition, Cx26 and Cx43 negative PTs developed Cx26 and Cx43 positive MLNs. Furthermore, besides increased cytoplasmic staining, enhanced membranous localisation of Cx43, typical of normal cells, was found in MLNs. Additionally, membranous Cx26 expression appeared only in metastatic breast cancer cells. CONCLUSIONS: These findings suggest that connexins may contribute to the efficient metastasising of breast cancer to the lymph nodes.

P53 correlates positively with VEGF in preoperative sera of colorectal cancer patients.

Diversity of P53 impact on tumor angiogenesis is due to the fact that wild-type P53 decreases expression of vascular endothelial growth factor (VEGF), but mutant P53 upregulates it. Therefore, we aimed at uncovering relations between preoperative serum levels of VEGF and P53 in colorectal cancer (CRC) patients. Preoperative blood samples of 125 CRC patients and 16 control healthy volunteers were examined with an ELISA-kit for serum P53 levels and VEGF. P53 did not correlate with VEGF in the whole group of CRC patients. However, P53 associated with VEGF in case of colorectal cancer patients, whose serum values of VEGF were higher than in controls (VEGF{H} >5.9333 pg/ml) (r=0.274, p<0.009). We revealed a positive correlation between P53 and VEGF{H} in subsets of poorly differentiated (G3) cancers (p<0.02), lymph node positive (p<0.007), pT3 or pT4 patients (p<0.004) without analogous relation in moderately differentiated (G2) tumors, node negative patients or pT1 or pT2 patients. P53 and IGF-I negatively correlated in all CRC patients (p<0.04) and VEGF{H} individuals of pT3 or pT4 (p<0.05) without any significant linkage in tumors of pT1 or pT2. The positive correlation between serum P53 and VEGF points at mutation of P53 and is a highly probable sign of poor prognosis in colorectal cancer. For now it can not be excluded that the binary analysis of serum P53 and VEGF could help select CRC patients endangered by rapid growth and lymph node metastases.

Expression of the Insulin Receptor Substrate 1 in primary tumors and lymph node metastases in breast cancer: correlations with Bcl-xL and Bax proteins.

In our previous investigation Insulin Receptor Substrate 1 (IRS-1) correlated with proliferation marker Ki-67 in human breast cancer. The aim of the present study was to assess relationships between IRS-1 expression and anti-apoptotic Bcl-xL as well as proapoptotic Bax proteins, assessed by immunohistochemistry, in primary tumors and lymph node metastases of breast cancer. IRS-1 is positively associated with both Bcl-xL and Bax in primary and metastatic tumors. Thus, our results could suggest that IRS-1 might affect turnover of cancer cells and breast cancer progression through activation of mitogenesis and participation in the regulation of the balance between anti- and proapoptotic pathways.

The role of Bak expression in apoptosis of the oral squamous cell carcinoma (OSCC) and metastases to lymph nodes (LNMs).

The immunohistochemical method was applied to show Bak expression in oral squamous cell carcinoma and its metastases to lymph nodes (LNMs). Bak expression was evaluated by immunohistochemical methods in specimens with oral squamous cell carcinomas and their lymph node metastases. Immunohistochemical studies were performed, using goat polyclonal Bak antibodies (Santa Cruz Biotechnology, USA) at 1:200 dilution. Our studies revealed over expression (64%) of Bak in the cytoplasm of epithelial cells in primary tumours (PTs) and in (75%) LNMs. No statistically significant correlations were observed between Bak immunoreactivity and age, pT and G of the carcinoma in PTs and LNMs. We conclude that expression of Bak may be useful for better characterising and predicting the prognosis of OSCC but cooperative studies are needed to assess its applications in the clinical practice.

Proliferating activity in the epithelial and stromal component of fibroadenomas and phyllodes tumours of the breast.

The aim of the study was an evaluation of PCNA and Ki-67 expression in the epithelial and stromal component of fibroepithelial tumours (FT) of the breast in correlation with morphological parameters. A series of 11 fibroadenomas (FA), including 8 cases of the cellular type (FAC), 19 benign phyllodes tumours (PTLGM), 8 bordeline (PTBM) and 6 malignant phyllodes tumours were assessed, using immunohistochemistry. The expressions of Ki-67 and PCNA in the epithelial component were significantly higher in PTLGM, when compared with FA and PTBM. A significant increase of Ki-67 and PCNA stromal expressions was associated with the progression from PTLGM to PTHGM. Our results show that Ki-67 and PCNA may be useful in the evaluation of stromal proliferation in phyllodes tumours (PT), which play an integral part in the progression from PTLGM through PTBM to PTHGM.

The Bcl-2 protein expression in germinal centers of hypertrophied adenoids in children.

The aim of this study was to evaluate Bcl-2 expression in the germinal centers of adenoid in children on the assumption that it can be treated as a marker of adenoidal tissue function. The study involved 95 children. The patients were divided into three age groups: aged up to 5, 5-10 and above 10 years. The analysed material were adenoids removed on the grounds of hypertrophy. Immunohistochemical analyses were carried out using monoclonal antibodies (DAKO/Bcl-2, No. M0887) directed against human Bcl-2 protein. The presence of Bcl-2 positive lymphocytes within the lymphoid follicles and Bcl-2 immunostaining were scored. The immunohistochemical staining showed Bcl-2 positive lymphocytes in the examined sections of adenoidal tissue and their characteristic location, mainly within the mantle zone. Bcl-2 reactivity was widespread in hyperplastic lymphoid tissue, including germinal centers surroundings, scattered cells within the germinal centers and the T-cell areas in general. We have not found statistically significant correlation between Bcl-2 expression and clinical status and change in Bcl-2 expression level according to age. The apoptosis presence within the germinal centers and its lack in mantle zone, the manifestation of which is Bcl-2 expression, is a sign of a proper lymphocytes maturation within lymphoid follicles. It seems that these processes are not influenced by age, so the adenoid involution is probably associated with the changes in the number of lymphoid follicles rather than change or inhibition of the processes that take place in them.

Chymotrypsin-like activity in rat tissues in experimental acute pancreatitis.

PURPOSE: Increase in intracellular chymotrypsin activity was reported during acute pancreatitis. Beside chymotrypsin, there are at least two enzymes with chymotrypsin-like activity: proteasome and lysosomal cathepsin A. Until now it is not known whether and to what extent they contribute to increases in chymotrypsin activity in acute pancreatitis. Our aim was to study organ chymotrypsin-like activities during experimental acute pancreatitis. MATERIAL AND METHODS: Rat cerulein model of acute pancreatitis was used. The chymotrypsin-like activities were assessed in pancreas, liver, lung, heart, spleen and kidney using highly selective synthetic substrates of the proteasome and the cathepsin A, at neutral and acidic pH. Determinations after addition of selective inhibitor were also performed. RESULTS: During acute pancreatitis we found in the pancreas an increase only in neutral chymotrypsin-like activity, as compared to the control animals. In other organs neutral chymotrypsin-like activity did not increase, and in kidney it even decreased. There were no changes in acidic chymotrypsin-like activity in any of organs studied. The studies using the inhibitor of the proteasome showed that the neutral chymotrypsin-like activity in the pancreas of the rats with acute pancreatitis should not be attributed to the proteasome activity, but rather to the chymotrypsin. CONCLUSIONS: Our results did not confirm any significant contribution of proteasome or cathepsin A to increased chymotrypsin-like activity in acute pancreatitis. We showed a decrease in neutral chymotrypsin-like activity of proteasome in the kidney, but the significance of this finding remains to be established.

[Malignant fibrous histiocytoma (fibrohistiocytoma malignum) localized intracranially]. / Wlókniako-miesak zlosliwy (fibrohistiocytoma malignum) o wewnatrzczaszkowej lokalizacji.

Primary intracranial malignant fibrous histiocytoma (MFH) is an extremely rare occurrence, however this kind of neoplasm is observed among the sarcomas. The aim of this report is to present a case of malignant fibrous histiocytoma in a 22 years-old woman. The control examination performed five years after operation revealed only mild exophthalmus and visual disturbances as presented before surgery.

Fetotoxic effect of 2,4-dichlorophenoxyacetic acid (2,4-D) in rats.

The fetotoxic effect of 2,4-dichlorophenoxyacetic acid (2,4-D) was investigated. Histological and histochemical changes in the liver of newborn, jung and adult rats exposed to the herbicide from the prenatal period to the end of an experiment were evaluated. The experiment used 90 male and female, Wistar, aged to 10 weeks rats, divided into two groups: I-control-30 and II-60 animals which received the water solution of 2,4-D acid sodium salt in a daily dose of 250 mg/kg b.w. It was given with drinking water every day. The animals were sacrificed after 24 hours, 4, 6, and 10 weeks of the experiment. The results obtained showed that the administration of 2,4-D acid to rats in the prenatal and postnatal period, in a dose inducing subacute intoxication leads to histological and histochemical changes in the liver. The observed changes indicate disorders in energetic processes in hepatocytes and are morphological exponents of detoxicative processes there. They are most intensified with newborn rats. It suggest also, the pregnants ought not to work with 2,4-D and should avoid any contact with herbicides belongs to the 2,4-dichlorophenoxyacetic acid group.

Ultrastructural changes in rat hepatocytes in acute intoxication with 2,4-dichlorophenoxyacetic acid (2,4-D).

The hepatotoxic effect of 2,4-dichlorophenoxyacetic acid (2,4-D) was investigated. Ultrastructural changes were evaluated in transmission electron microscope. The experiment used 60 male Wistar rats divided into two groups: I--control-18 animals and II--42 animals which received chemically pure 2,4-D acid by gastric gavage in a dose of 200 mg/kg b.w. The animals were sacrificed after 12, 24, 48 hours and 4, 10 and 30 days of the experiment. The results obtained indicate that the administration of 2,4-D acid to rats in a dose inducing acute intoxication leads to ultrastructural changes in the liver, which suggest nonspecific reversible adaptative-type damage to parenchymal cells. The changes observed indicate disorders in energetic processes in hepatocytes and are morphological exponents of detoxicative processes there.

Morphological changes in mitochondria and lysosomes of hepatocytes in acute intoxication with 2,4-dichlorophenoxyacetic acid (2,4-D).

The hepatotoxic effect of 2,4-dichlorophenoxyacetic acid (2,4-D) was investigated. Ultrastructural changes were evaluated under a transmission electron microscope. Certain histoenzymatic reactions were examined (to acid phosphatase according to Gomori (AP) and to succinic dehydrogenase (SD) according to Nachlas) in parenchymal cells of the rat liver in acute intoxication induced by this herbicide. The experiment used 60 male Wistar rats divided into two groups: I-control--18 animals and II--42 animals which received chemically pure 2,4-D acid by gastric gavage in a dose of 200 mg/kg b.w. The animals were sacrificed after 12, 24, 48 hours and 4, 10 and 30 days of the experiment. The results indicate that the administration of 2,4-D acid to rats in a dose inducing acute intoxication leads to histoenzymatic and ultrastructural changes in the liver, which suggest nonspecific reversible adaptive-type damage to parenchymal cells. The changes observed indicate disorders in energetic processes in hepatocytes and are morphological exponents of intense detoxicative processes.

Primary intracranial malignant fibrous histiocytoma--ultrastructural study.

Malignant Fibrous Histiocytoma is the most common soft tissue sarcoma in adults with the majority of cases that occur in patients between 50-70 years and 64% of cases occurs in male. Most commonly it arises in extremities (lower 49% and upper 19%), less often in the abdomen (16%) and very rare in head localization (3%) [5]. Typically macroscopically it is a multilobulated, gray-white, fleshy mass between 5-20 cm of diameter. In light microscopy this tumour has a highly variable morphologic pattern and manifests a broad range of histological appearance. It has been classified into the following subtypes: storiform-pleomorphic, myxoid, giant cell, inflammatory and angiomatoid. The first two subtypes are more frequent than others. The aim of this report is to present the electronmicroscopic features of giant cell type of MFH localized in head.

Cyclophosphamide in diffuse lung damage.

Some cyclophosphamide toxic effects on lung tissue are presented. Cyclophosphamide metabolism, pathogenesis of lung damage and morphological lung tissue changes caused by that agent were characterized. Attention was focused on BAL evaluation as a useful method in the monitoring of lung tissue damage degree.

Cyclophosphamide-induced changes of serum angiotensin converting enzyme activity and pulmonary microvessels ultrastructure.

The effect of cyclophosphamide (CP) on the ultrastructure of the lung tissue and the activity of angiotensin converting enzyme (ACE) in serum was evaluated in rats. The animals were given cyclophosphamide (CP) in a single intraperitoneal dose of 150 mg/kg b.w. ACE activity was evaluated in the blood serum collected from the left ventricle of the heart using the spectrophometric method. In all time subgroups, the CP-receiving animals showed a decrease in ACE activity. Ultrastructural examinations of CP-treated animals revealed increased adhesion of neutrophiles and monocytes to the damage endothelium of the alveolar septa vessels and focally accumulation of the platelets.

Effects of pentoxifylline on some peripheral blood parameters and haemostasis in acute pulmonary tissue injury after cyclophosphamide.

Cyclophosphamide (CP) is one of the widely used cytostatic drugs, with a strong toxic influence on pulmonary tissue. Experimental works have shown that a single high dose of CP causes injury to all elements of the interalveolar septum, especially to the alveolar endothelial and epithelial cells. The aim of this work was to evaluate the influence of pentoxifylline (PTXF) (30 mg/kg b.w.) on the ultrastructure of lungs capillaries and blood cell count as well as plasma fibrinogen levels in Wistar rats after intraperitoneal injection of 150 mg/b.w. CP. We established that in the doses applied PTXF had no statistically significant influence on the number of leukocytes and erythrocytes determined in the left ventricular blood of rats receiving CP, while the number of white cells from animals given PTXF only was higher than in controls. There was a smaller decrease in the number of platelets (p < 0.05) and smaller reduction in fibrinogen level (p < 0.01) in the serum of PTXF-CP animals than in the CP group. The results obtained suggest a protective effect of PTXF on CP induced changes, which have been evidenced in some of the parameters examined. Ultrastructural examinations found the lungs to be the organ of extramedullary thrombocytopoiesis in CP-treated animals and revealed that platelet accumulation in the system of lung capillaries was a potential cause of the decrease observed in the number of blood platelets following CP administration.

Ultrastructure of bone marrow megakaryocytes in experimental haemorrhagic shock in rats. I. Correlation between ultrastructure of MK nuclei and DNA ploidy.

The correlation was described between the ultrastructural picture and DNA mass content of MK nuclei in experimental haemorrhagic shock in rats. Significant disproportions were revealed between the morphological pictures and the ploidy of MK nuclei in the successive phases of the shock. Abnormalities of the maturation of marrow MK nuclei were found in the first hours of the shock, being most pronounced in the 24th hour.

Morphometric and ultrastructural studies of the spinal sensory ganglia in the course of sciatic nerve regeneration in rats intoxicated with ethanol.

The aim of the study was to examine morphological changes in the spinal dorsal ganglia (from which sensory fibres of the sciatic nerve extend) in animals intoxicated with ethanol, in which the sciatic nerve was transected and then sutured. By means of morphometric methods using a computer system of picture analysis and in ultrastructural examinations it was found that ethanol administered to rats to drink for 3 months caused only slight morphological alterations in the sensory nerve cells of the spinal vertebral ganglia on the non-operated side. However, ethanol or its metabolites destroyed the regenerating sensory nerve cells of the spinal vertebral ganglia of the transected and then sutured sciatic nerve.

Fine needle aspiration biopsy cytology of pulmonary tumors.

Transthoracic fine needle aspiration specimens of the pulmonary tumors were obtained from 144 patients. Fine needle aspiration biopsy were performed using local anaesthesia and ultrasonographic or scopie control. The 20-gauge needles were used to obtain the specimens. 93 of patients have been diagnosed as squamous cell carcinomas, 28 as adenoid carcinomas, 18 as anaplastic carcinomas and 5 as a non neoplastic lesions (tuberculoma and abscessus). The tumors were typed according to the second WHO histological classification. Analysis of the date indicated that malignant neoplasms were identified correctly with an accuracy of 92.0%. There were not false positive diagnoses. There were two false negative diagnoses (squamous cell carcinoma) and in three cases the diagnosis were as suspected for malignancy. The results confirmed the value of fine needle aspiration cytopathology for the diagnosis of pulmonary tumors.

The ethanol effect on lung metastases development in experimental Morris 5123 hepatoma.

The experiment was carried out on 45 male Wistar rats which were divided into 3 groups: group I-15 rats which were given water to drink during the experiment, group II-15 rats obtaining 10% ethanol solution during the experiment and group III-15 rats obtaining 20% ethanol solution during the experiment. All animals were injected 0.3 ml suspension of hepatoma Morris 5123 cells directly to the liver on the 14th day of the experiment. After the 9 weeks of the experiment the animals from all groups were narcotized, decapitated and the lungs were taken into the morphometric, histological and ultrastructural examinations. They showed that ethanol has a stimulating effect on formation and development of hepatoma Morris 5123 in rat lungs. The increase of number and extensiveness of metastases as well as the increase of mean metastases focuses volume, and the increase of lung weight in animals which obtained ethanol are the exponent of the influence. On the basis of ultrastructural examinations, it can be noted that promoting activity of ethanol can be connected: a) with the increase of neoplastic cells activity, b) with changes appearing in pulmonic epithelium and endothelium of vessel enabling neoplastic cells to adhere to basal membrane as well as their crossing the vessel wall.

[Intracranial chordomas; histochemical and immunohistochemical examinations]. / Struniaki wewnatrzczaszkowe; badania histochemiczne i immunohistochemiczne.

Chordoma is one of rare intracranial neoplasms (0.2-0.5%). Apart from classic chordoma in 1973 r. Heffelfinger and al. separated chondroid chordoma which showed better prognosis. We studied 4 classic and 3 chondroid intracranial chordomas. All tumours contained chondroitine sulfate and keratan sulfate and showed positive immunohistochemical reactions to cytokeratin, S-100 protein and vimentin, supporting the concept of common origin of both types of chordomas. Immunohistochemical studies of PCNA and Ki-67 antigen and staining AgNOR's did not show significant differences in the proliferative activity between both types chordomas what attests to biological malignancy.