Local inflammatory response in colorectal cancer.

Type and intensity of tumor-infiltrating lymphocytes (TILs) in close proximity to the primary tumor are prognostically significant in postoperative patients. High intensity of TILs is considered to be a prognostically beneficial factor. The research included 66 postoperative colorectal cancer patients. The control group comprised 20 colon segments. Monoclonal antibodies LCA, CD3, CD4, CD5, CD8, CD20, CD23 and CD138 were used to differentiate between T and B lymphocytes. Types of cells in the infiltrate were defined. We found greater numbers of T and B lymphocytes located in close proximity to the cancerous tissue when compared to the control group. T lymphocyte intensity in the inflammatory infiltrations was directly correlated with the size of resected tumors, presence of regional lymphatic node metastases and histological grade of malignancy. Lymphocytic infiltrations of greater intensity located in close proximity to the primary tumor were found in subjects with less advanced colorectal cancer. The research presented here proves direct dependence between the immune system and colorectal cancer. The presence of lymphocytes in the inflammatory infiltrations located in close proximity to the cancerous tissue has been proved to be prognostically beneficial. The obtained results support the application of immunotherapy in colorectal cancer treatment.

The impact of TNF superfamily molecules on overall survival in acute myeloid leukaemia: correlation with biological and clinical features.

B cell-activating factor (BAFF), a proliferation-inducing ligand (APRIL) and apoptosis-inducing ligand (TRAIL) were demonstrated in several haematological diseases including acute myeloid leukemia (AML). Those cytokines are capable of activating a broad spectrum of intracellular signalling cascades that can either induce apoptosis or protect from programmed cell death. We have analysed BAFF, APRIL and TRAIL serum concentrations in 76 patients with newly diagnosed AML and 40 healthy volunteers. The values were significantly higher for APRIL and BAFF but lower for TRAIL compared to healthy volunteers. Induction therapy significantly reduced the values for BAFF and increased them for TRAIL. Moreover, the concentration of BAFF and APRIL was significantly lower and the concentration of TRAIL higher in a group of patients with complete remission compared to non-respondent AML patients. In addition, higher concentrations of BAFF and lower of TRAIL predicted a shorter overall survival, suggesting thereby an important prognostic marker and possible therapeutic target in AML.

BAFF and APRIL as TNF superfamily molecules and angiogenesis parallel progression of human multiple myeloma.

Tumour necrosis factor alpha (TNF-α) is an inflammatory cytokine with a wide spectrum of biological activity, including angiogenesis. B cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are members of the TNF-α family. Vascular endothelial growth factor (VEGF), on the other hand, is one of the most characteristic pro-angiogenic cytokines produced by multiple cell types in multiple myeloma (MM). We have analysed BAFF and APRIL concentrations in parallel with pro-angiogenic cytokines in serum and trephine biopsy, and the bone marrow microvascular density (MVD) in 50 patients with newly diagnosed IgG MM and 24 healthy volunteers. The study showed statistically higher concentrations of BAFF, APRIL and TNF-α, as well as VEGF and its receptor, in MM patients compared to healthy volunteers and patients in advanced stages of the disease. A statistically positive correlation between the concentration of TNF-α and the expression of VEGF was demonstrated, and so was a positive link between BAFF, APRIL, MVD and lactate dehydrogenase (LDH). Furthermore, we observed a significant decrease in all studied cytokines after anti-angiogenic therapy, with meaningful differences between responders (at least partial remission) and patients with stable disease. It was also established that APRIL, but not BAFF, correlated with pro-angiogenic cytokines such as VEGF with its receptor, MVD and syndecan-1. Finally, our results showed that serum BAFF and APRIL levels could be useful biomarkers of MM disease activity and its progression which suggests that APRIL could be a possible novel therapeutic target in MM.

Imidazole-induced contractility of vascular smooth muscle cells in the presence of U-73122, ODQ, indomethacin and 7-nitroindazole.

The aim of the study was to assess the impact of modulating factors on vascular smooth muscle cells reactivity. Vascular resistance was induced by the administration of increasing concentrations of imidazole. The experiments were performed on isolated and perfused tail artery of Wistar rats (weight 250 g - 350 g). Rats were been narcotized by urethane (intraperitoneal injection) at a dose of 120 mg/kg, stunned and then sacrificed by cervical dislocation. In the following investigation classical pharmacometric methods were used. Relationships between concentration-response curves (CRCs) for imidazole observed in the presence of ODQ [(1H-(1,2,4)oxadiazolo-[4,3-a]quinoxalin-1-one)], 7-nitroindazole and indomethacin were analyzed. Imidazole-induced contractility of vascular smooth muscle cells was independent from alpha-adrenergic receptors and PLC activity. Reactivity of VSMCsinduced by imidazole, was significantly changed in the presence of ODQ and 7-nitroindazole.

Lung mass in a 28-year-old male: a case report of a rare tumor.

A twenty eight-year-old male presented with a two week history of dyspnea, cough, hemoptysis, chest pain, and fever 38-39°C. He also complained of loss of appetite, general weakness and left leg pain for two months preceding admission. He was referred with suspicion of lung tumor to our institution. Chest X-ray showed almost total atelectasis of the right lung with compensatory overinflation of the contralateral lung. Using computed tomography (CT), a lesion of diameter of 19.3 x 14.1 x 19.1 cm in the right lung, pleuritis, Th3 osteolysis, and compensatory overinflation of the left lung was seen. Bronchoscopy revealed a total obstruction of the right main bronchus due to submucosal infiltration and compression of the right main bronchus with negative histology of bronchial biopsy specimens. Transthoracic fine needle aspiration revealed celullae suspectae probabiliter neoplasmaticae suggesting tumor fusocellularis. USG of the abdomen revealed liver with numerous heterogeneous, solid areas hypo- and hyperechogenic, some of them with features of liquid or the disintegration up to diameter of 74 mm. Subsequent fine needle aspirations of the thorax and liver revealed fibrolamellar hepatocarcinoma and carcinoma adenoides of the lung. Patient underwent chemotherapy with 5-FU/DDP/VCR with no response. This report presents a case of a rare lung metastasis from FL-HCC.

Prognostic significance of ligands belonging to tumour necrosis factor superfamily in acute lymphoblastic leukaemia.

Altered activities of ligands belonging to tumour necrosis factor (TNF) superfamily, namely B-cell activating factor (BAFF), a proliferation-inducing ligand (APRIL) and apoptosis inducing ligand (TRAIL) were demonstrated in several haematological diseases including acute lymphoblastic leukaemia (ALL). BAFF, APRIL and TRAIL provide crucial survival signals to immature, naive and activated B cells. These ligands are capable of activating a broad spectrum of intracellular signalling cascades that can either induce apoptosis or protect from programmed cell death. BAFF and APRIL, which can directly activate the NF-κB pathway, have been identified as crucial survival factors for ALL cells. Here, we have analyzed serum BAFF, APRIL and TRAIL concentrations in 48 patients with newly diagnosed ALL and 44 healthy volunteers. The levels of APRIL and BAFF were significantly higher in ALL patients as compared to healthy volunteers. In contrast, concentrations of TRAIL were significantly lower in ALL patients. Moreover, following induction, the levels of APRIL, but not BAFF or TRAIL, were significantly lower in a group of patients with complete remission (CR) as compared to non-respondent (NR) ALL patients. Furthermore, we demonstrated statistically significant differences in concentrations of APRIL between CR MRD-negative and CR, MRD-positive ALL patients. Notably detection of higher concentrations of APRIL was associated with shorter leukaemia-free survival and overall survival. Altogether, our data indicate that APRIL can play an important role in the pathogenesis of ALL and the measurement of APRIL levels can improve prognostication in ALL patients.

The role of nitric oxide in caerulein-induced acute pancreatitis and the recovery process after inflammatory damage.

OBJECTIVES: Nitric oxide (NO) is involved in the control of pancreatic blood flow and secretion, and its role in the maintenance of pancreatic tissue has been suggested. The aim of our study was to evaluate the influence of NO on pancreatic trophic parameters during acute pancreatitis induction and the pancreas recovery process. DESIGN/METHODS: Acute pancreatitis was induced in rats by a supramaximal dose of caerulein. During acute pancreatitis induction, rats were treated with L-arginine (a substrate for NO synthesis), glyceryl trinitrate (NTG, NO donor), glycine, N(G)-nitro-L-arginine (L-NNA, NO synthase inhibitor), L-arginine + L-NNA, or saline. Pancreatic weight, total contents of RNA, DNA, protein, amylase and chymotrypsin as well as pancreas histology and the number of proliferating acinar cells were evaluated after pancreatitis induction in all groups and additionally after 7 and 14 days of recovery in untreated acute pancreatitis rats or those injected with L-NNA and/or L-arginine. RESULTS: Pancreas destruction after acute pancreatitis was evidenced by similar decreases of all parameters in untreated acute pancreatitis rats or those treated with L-NNA or glycine when compared to control healthy animals. The recovery process after acute pancreatitis was not affected by L-NNA injections; however, the increased cell proliferation occurred later than in untreated rats. NTG and especially L-arginine treatment resulted in significant attenuation of pancreas damage (partially prevented by L-NNA treatment) as evidenced by biochemical data with a slight improvement in morphology. Treatment with L-arginine alone or in combination with L-NNA resulted in augmented cell proliferation after acute pancreatitis induction followed by earlier recovery in comparison to untreated acute pancreatitis rats or the L-NNA-injected group. CONCLUSION: The present study suggests the involvement of NO in mild acute pancreatitis and in the recovery process after inflammatory damage.

Does chronic ethanol administration have influence on pancreatic regeneration in the course of caerulein induced acute pancreatitis in rats.

UNLABELLED: This study was undertaken in order to determine the influence of chronic ethanol administration on pancreatic regeneration during acute pancreatitis (AP). Rats were pair fed with isocaloric diet containing or not ethanol. After 8 weeks of such feeding AP was induced by s.c. injection of caerulein (Cae). 6 h, 24 h and 5 days after first Cae dose pancreatic weight, amylase, chymotrypsin, protein, RNA, DNA contents were determined and phosphatidic acid (PA) production in isolated pancreatic acini was measured. Proliferating cells were quantified by immunochemical staining of cells incorporating bromodeoxyuridine (BrdU). RESULTS: Pancreatic weight was significantly higher at 6 h after first Cae injection in both, ethanol fed (EF) and control groups (C), however at 24 h pancreatic weight did not differ from prior to AP induction in EF rats. Ethanol feeding (EF) did not influence significantly protein, chymotrypsin and amylase content in pancreatic tissue in groups with AP. In EF rats RNA content after 5 days of AP was higher than in control animals. Total DNA content in EF rats with AP was lower 6 h after AP induction, earlier than in control animals with AP. Immunochemistry showed higher labelling index for BrdU after 6 h, 24 h and 5 days of AP in EF rats. In contrast to this findings, in EF animals, AP induction was not able to stimulate further PA accumulation. CONCLUSION: We conclude that chronic ethanol feeding, while inhibiting PA accumulation in comparison to control group, does not impair pancreatic tissue regeneration during the early phase of Cae-induced AP. Stimulation of regenerative/reparative processes in EF rats during Cae-induced AP seems to be even more pronounced than in the control group.

The influence of glyco-nitric oxide conjugate on proliferation of breast cancer cells in vitro.

The aim of the study was to evaluate the influence of a nitric oxide donor: Glyco-2-SNAP on the proliferation status of two breast cancer cell lines: MDA-MB-231 and MCF-7. The study was performed by the thymidine incorporation method as well as by the immunocytochemical detection of the Ki-67 antigen. We found that the donor significantly inhibits the process of proliferation. The effect of the Glyco-2-SNAP is significant in both lines, however stronger in MDA -MB-231 cells where the donor at 100 microM inhibited DNA synthesis from 70462.000 dpi (SD +/-2066.175, n=4) for control to about 3120.250 dpi (SD +/-971.689 n=4). In UCF-7 cells the control gave 31142.500 dpi (SD +/-712.9214, n=4) and the treatment with 100 microM of Glyco-2 SNAP resulted in 4095.50 dpi. (SD +/-315.723, n=4). In both lines SNAP, classical NO donor also had an inhibitory effect but much higher concentrations. Ki-67 protein expression was significantly influenced by Glyco-2-SNAP at 100 microM (42.5% +/-6.45) concentrations in MDA-MB 231. No effect of G-SNAP was seen in MCF-7 cells. The control staining for both lines was about 90%. Our results presesent the possibility of exploiting a novel kind of NO donor as a potential mode of treatment that may be an alternative to classical therapeutic strategies in breast cancer.

Megakaryocytes emperipolesis in bone marrow of the patients with non-Hodgkin's lymphoma.

Emperipolesis of hematopoietic cells within the cytoplasm of the megakaryocytes was most often described in association with various pathologic conditions. The aim of the research was estimation of the incidence of emperipolesis in the bone marrow of the patients with non-Hodgkin's lymphoma (NHL). 30 patients with different histological types of NHL (chronic lymphocytic leukaemia--CLL, hairy cell leukaemia--HCL, multiple myeloma--MM) in compliance with clinical stage of the disease, patient's age and sex were analyzed. Trephine biopsies of the bone marrow were carried out in fixative solution and paraffin embedding. Hematoxylin and eosin and monoclonal antibody CD 61 were applied on thin sections. Phenomenon of megakaryocytic emperipolesis in human bone marrow was found in 6 cases: in 5 cases in CLL and in 1 case in HCL. In most of them emperipolesis was related to single megakaryocytes. We observed in the cytoplasm of the megakaryocytes single hematopietic cells-most often lymphocytes, rarely eosinophilic granulocytes. We found no correlations between histological types of NHL, clinical stage of the disease, patients' age, sex and the incidence of megakaryocytic emperipolesis.

Proliferative activity of the thyroid oxyphilic tumor cells estimated by means of quantitative analysis of silver stained nucleolar organizer regions (AgNORs).

UNLABELLED: Promising results of studies on different neoplasms, by means of morphometric analysis of argyrophilic nucleolar organizer regions (AgNORs), known as proliferative activity marker, made us undertake an attempt to evaluate of proliferative activity in Hürthle Cell Tumors using the same technique. 78 cases including 20 Hürthle Cell Carcinomas (HCC), 32 Hürthle Cell Adenomas (HCA) and 26 hyperplastic nodules with Hürthle Cell Metaplasia (HCM), diagnosed in the Department of Clinical Pathology, Medical Academy of Bialystok in the years 1990-2000, were subjected to analysis. For visualization of the NORs we used the D. Ploton et al. technique. Mean AgNOR count and NORDS (NOR distribution score--the percentage of nuclei with at least 5 argyrophilic granules) were estimated in each case in 100 randomly chosen nuclei. Non-parametric Mann-Whitney U-test was applied to determine statistically significant differences. RESULTS: Mean AgNoRs counts and NORDS values were 5.1 and 52%, 3.4 and 26%, 2.5 and 7% in HCC, HCA and HCM respectively. Statistically significant differences were found between AgNORs counts in carcinomas and adenomas (p<0.01), HCCs and HCMs (p<0.005) and betwveen NORDS in all groups (p<0.001).

Protein kinase C involvement in proliferation and survival of breast cancer cells.

Members of protein kinase C (PKC) family have been widely implicated in the regulation of cell proliferation, differentiation and survival. Increased protein C activity in malignant breast tissue and in most aggressive breast cancer cell lines suggests possible role of PKC in the development and progression of breast cancer. PKC may be therefore a target for breast cancer treatment. In our study we attempted to investigate the effect of: phorbol ester (PMA)-PKC activator, and bisindolylmaleimide II (GF II), a highly selective PKC inhibitor, on the proliferation as well as induction of apoptosis and necrosis in breast cancer cell line MDA-MB-231. Our results provide evidence for multidirectional effects of PKC on the proliferation of this type of breast cancer cells. The effects of both compounds were different after short time of exposition (1-3 h). PMA induced proliferation, while GF II showed an opposite effect. After 24 h, however, both compounds exhibited relatively high inhibitory effect on the proliferation and proved to be effective in induction of necrosis and apoptosis.

Influence of brain hypoxia on megakaryocytic emperipolesis in rats.

An evaluation of frequency and intensity of the megakaryocytic emperipolesis in the rat bone marrow after unilateral carotid artery occlusion was performed. The results were compared with those obtained in the group undergoing a blood loss and the group not subjected to surgical procedures. A significant increase in the frequency of emperipolesis was found in the bled rats following a carotid artery cannulation, but also in animals exposed to the carotid artery occlusion only. In most cases the cytoplasm of megakaryocytes contained granulocytes and erythroblasts. The observed emperipolesis phenomenon is likely to be a result of brain hypoxia and a secondary neurogenic stimulation of the bone marrow activity.

MIB-I as a proliferative activity marker of the multiform glioblastomas.

MIB-I is a proliferative activity marker of multiform glioblastomas which are the most frequent tumors of the central nervous system. They are characterizad by differential rate and prognosis. The aim of the study was to determine the proliferative activity of multiform glioblastomas and estimation of the correlation between tumors' proliferative activity and tumors' localization, size, patients' age and sex. 24 patients (18 females and 6 males) with multiform glioblastomas were analyzed. The mean patients' age was 52.1. The proliferative activity was calculated as a proliferation index: IP for MIB-I. Cells with positive reaction were determined by MIB-I which was compared to all neoplastic cells. The most frequent localization of the tumors were frontal and temporal lobes of the brain. The size of the tumors ranged from 2.5 to 5.3 cm (mean 3.9). Mean IP was 43.2 (SD+/-17.4). We found no correlation between IP MIB-I and localization of the tumor, patients' age and sex. There was a marginal statistically significant correlation between IP MIB-I and size of the tumor (p=0.005).

Proliferative activity of papillary carcinoma and benign papillary hyperplasia of thyroid follicular cells.

Papillary structures of follicular cells are observed in nodular goiter, cysts, hyperplastic areas of follicular tumors, Graves' disease, thyroiditis and carcinomas. The distinction of papillary carcinomas from benign lesions has important implication for clinical management. The aim of the study was to test a marker of proliferation activity (MIB-1) in the diagnosis of benign and malignant thyroid papillary proliferation. The study was carried out in 98 women with papillary carcinoma, nodular goiter. intracystic proliferation. Graves' disease and hyperplastic areas of follicular benign tumors. The formalin fixed, paraffin-embedded specimens were microscopically examined using HE staining and immunostaining with MIB-1 antibody (DAKO). The proliferative index (PI) was significantly higher in malignant than in benign papillary hyperplasia. Our results may provide additional information for differential papillary proliferation diagnosis by FNAB.

Preliminary immunohistochemical investigations of human thyroid C cells in the simple and hyperactive nodular goitre.

The aim of this study was to carry out histomorphological and immunohistochemical analysis of thyroid C cells in 30 patients with simple goitre and hyperactive goitre including Graves-Basedow (G-B) disease, treated surgically. Four tissue samples were always taken from the same internal parts of the gland where the number of C cells in physiological conditions was the highest. C cells were detected in paraffin sections after impregnation with silver salt (Grimelius method) or immunohistochemically, with antibodies against calcitonin, synaptophysin, chromogranin A and neuron-specific enolase. Distinctly less numerous C cells were found in simple and hyperactive goitre than in normal thyroid parenchyma. The majority of C cells showed weak intensity of the examined immunohistochemical reactions. C cells were not observed at all in the texture of nodules well separated by the connective tissue. Proliferative changes concerned only follicular cells.

Preliminary evaluation of influence of gemcitabine (Gemzar) on proliferation and neuroendocrine activity of human TT cell line: immunocytochemical investigations.

The choice treatment of medullary thyroid carcinoma (MTC) is total thyroidectomy. It is difficult to evaluate effectiveness of chemotherapy due to the rare incidence of MTC. Gemcitabine is a new drug of antimetabolite nucleoside group used in treatment of cancers since 1996. The aim of this study was to evaluate the influence of gemcitabine on proliferation and neuroendocrine activity of human TT cell line derived from MTC. The cells were exposed to gemcitabine in the concentration of 10, 25 and 50 microg/ml for 24 hours. Immunocytochemical examinations were carried out by the method of avidin-biotin peroxidase complex (ABC) according to Hsu et al. to detect calcitonin, chromogranin A, synaptophysin and neuron-specific enolase in TT cells. A concentration-dependent inhibitory influence of gemcitabine on proliferative activity of TT cells was observed. It was also shown that the immunostaining was reduced, especially in case of neuron-specific enolase. Only the reaction detecting calcitonin was enhanced in persistin

Fine needle aspiration cytology of benign breast disease. Markers of apoptosis and proliferation.

Aim of the study was to compare the fine needle aspiration cytology findings of benign breast lesions with incidence of proliferation markers and apoptosis. This study included 37 patients with palpable breast lumps, referred for USG guided FNA. FNAC were prospectively classified as C2-benign, C4-suspicious of malignancy, and C5-malignant. The specimens were simultaneously stained for Ki-67, MPM2, Bcl2 and P53. The diagnoses in group-C2 were following: simple cyst, multiple cysts, simple cyst with apocrine metaplasia, inflammatory cyst, benign dysplasia (BD) and benign solid tumors. The final diagnoses, after histopathological verification, in cases of primary classification as C4 and C5 were as follow: proliferative fibroadenoma (FAp) and breas cancer, respectively. Great majority of C2/BD aspirates were negative for proliferative antigens Ki-67 and PCNA. These antigens were detected in part of benign solid tumors, as anticipated in suspicious solid tumor, and in all of cancer aspirates. Bcl-2 immunopositive cells were detected approximately in one quarter of C2/BD, nearly in half of C2 solid tumors and in one C4/FAp. Most of diagnosed specimens were P53-negative. Immunocytodetection of Ki67, MPM2, Bcl2, P53 might be promising, supportive method in the classification of benign breast lesions. FNAC increases the reliability of diagnosis when complemented by immunocytochemical staining. It could be helpful procedure of establishing more accurately the biology of these lesions and possibly serve as an essential factor in clinical follow-up. Nevertheless, further study on larger group of patients comparing cytological and histopathological diagnosis is required to estimate reliability of its predictive value.

Influence of estrogen, antiestrogen and UV-light on the balance between proliferation and apoptosis in MCF-7 breast adenocarcinoma cells culture.

Studies of the mechanism of actions of estrogen, antiestrogen and physical factors may provide clues to an understanding of breast cancer growth and/or regression regulation and thus identify novel targets for therapeutic intervention. Defective control of apoptosis appears to play a central role in the pathogenesis of neoplasia. Conversely, cancer therapy and ionizing radiation can induce cancer cell death by apoptosis and/or necrosis. bcl-2 gene and p-53 gene products have been both linked to programmed cell death pathways. We have analyzed the effect of estradiol, tamoxifen and UV exposure on the induction of apoptosis, expression of p53 and bcl-2 gene products as well as the proliferative activity (expressed as [3H]thymidine incorporation and PCNA and MPM2 antigens involvement) in MCF7. It has been found that estradiol increases the speed of cell cycle in MCF7 and acts as antiapoptotic factor. Tamoxifen has multiple influence on the rate of growth of cancer cells: depends on estrogen receptor (ER), conducts reduction of proliferation rate; depends on ER and other mechanisms conducts to suppressions of Bcl-2 protein expression and induction of cell death through apoptotic pathway. Estradiol prevents the apoptotic influence of tamoxifen probably by enhancement of Bcl-2 protein expression and does not prevent the inhibition of proliferation rate. The irradiation with UV induces apoptosis by over-expression of p53 and down-regulation of bcl-2 gene.

Widespread myocardial and pulmonary bone marrow embolism following cardiac massage.

Widespread fatal massive bone marrow embolism of the coronary and the pulmonary arteries are described in a 71-year-old man with a respiratory disease. Cardiac massage was carried out during the cardiac arrest. In autopsy findings there were no fractures of the ribs or the sternum. The authors suggested that the cardiac massage was the most important predisposing factor for the bone marrow embolism.