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The aim of the study was to determine whether the character of a histopathological change observed in the epididymis of a 13-year-old European bison male was neoplastic, inflammatory or proliferous. Samples from both testes and epididymes were collected. The material was fixed in Bouin’s fluid; 6 mm paraffin sections were stained with hematoxylin and eosin (H+E), as well as by Brachet’s method. Full blood samples were collected from the femoral artery (post mortem). Immunocytochemical reactions were performed by the EnVisionTM FLEX+ technique. Specific monoclonal mouse and polyclonal rabbit antibodies were used against: vimentin (No IS630); human cytokeratin (No IS053); human CD 34 (No IS632); human desmin (No IS606); human smooth muscle myosin heavy chain (No IS066); myogenin (No IS067); human muscle actin (No M0635) in a 1:50 dilution; human MyoD1 (No M3512) in a 1:50 dilution; human myeloid/histiocyte (No M0747) in a 1:100 dilution; human receptor Cd 68 (No IS609); S-100 (No IS504). Micromorphometric measurements were carried out to assess the extent of connective tissue hyperplasia in the epididymis. The histopathological change in the epididymis of the 13-year-old bull was progressive and caused by chronic inflammation, which was confirmed by the micromorphometric analysis and the evidence of plasma cells and connective tissue hyperplasia. Immunohistochemical staining carried out to find vimentine receptors showed mesenchymatic cells forming disseminated foci in the epididymis. The appearance of the cells that made up the microgranulomas indicated their epithelioid nature. These transformed macrophages were devoid of the typical receptors Cd68 and Mac 387. However, the presence of spermatozoa in the foci suggested their phagocytic properties. Damage to the canaliculi of the epididymis and their phagocytosis could have been caused by an unknown proinflammatory factor. It is also likely that the destruction was induced by epithelioid cell proteins, which resulted in a dislocation of sperm to the epididymal stroma.
The aim of the study was to establish to what degree the 24-week exposure of a rat to 5 and 50 mg Cd/dm³ affects the proliferating activity of cells with PCNA and Ki-67 nuclear immunoreactivity in the submandibular gland cells. The control animals received only redistilled water to drink. The group I rats were given 5 mg Cd/dm³, while the group II animals were given 50 mg Cd/dm³. The highest concentration of cadmium was observed in group II, with a concomitant increase in the number of PCNA-positive cells. In group I, cadmium concentration was significantly less compared to group II, and there were fewer PCNA-positive cells. The reaction for Ki-67 in both experimental groups was negative.
In chronic lymphocytic leukaemia (CLL) bone marrow trephine biopsy (BMT) is not required for diagnosis but can have a significant prognostic value and can be used for the detection of the minimal residual disease (MRD) and for assessment of the effectiveness of the treatment applied. The aim of the study was to evaluate the morphological changes in bone marrow after treatment with purine nucleoside analogues cladribine and fludarabine. Bone marrow trephine biopsy was taken routinely from 15 patients with CLL. Bone marrow trephine biopsy was performed on every patient before as well as after chemotherapy. The number of cell elements of the marrow (the degree of atrophy), the patterns of bone marrow infiltration, the presence of reticulin and collagen fibres and the disturbances in bone marrow stroma were assessed. The infiltration of bone marrow by neoplastic cells was observed in all the patients before administration of chemotherapy. The infiltration was followed by an increase in the number of reticulin fibres. After the treatment a regression of the reticulin fibres was observed with the lessening of the infiltration. After the treatment the levels of marrow infiltrate were decreased. Increased hypoplasia of the bone marrow was observed after the chemotherapy.
The enlargement of the thyroid is, in general, benign in origin and due to nodular goitre. Follicular cellular proliferation of thyroid nodules has been increasingly observed recently. With fine needle aspiration biopsy (FNAB) this is classified as a follicular tumour. These lesions present various patterns of vascularisation in ultrasound examination. The aim of the study was to establish the relation between follicular nodule vascularisation and the proliferative activity of various types of follicular cell. According to the manner of proliferation, patients were divided into groups as follows: (I) patients with hyperplastic nodules (46 cases), (II) patients with follicular adenoma (42 cases), and (III) patients with follicular cancer (9 cases). In each case B-mode sonography, Power Doppler, sonographically guided FNAB (S-FNAB), morphological examination and morphometry were performed. The proliferative activity was detected with immunohistochemical methods (PCNA, Ki 67 and MPM2) to determine the so-called “proliferative index”. The study revealed increased proliferative activity in tumours of malignant origin and increased vascularisation in coexistence with increased proliferation of the follicular cells. As assessed by Power Doppler, an increased flow pattern in the centre of the nodules correlates with increased proliferative activity. The results suggest that Power Doppler examination could be helpful in selecting nodules for FNAB, especially in multinodular goitre.
The aim of the study was an estimation of the histomorphometry of megakaryocytes (MK) in bone marrow in selected myeloproliferative and lymphoproliferative diseases. Bone marrow specimens were obtained by trephine biopsies from 41 patients with polycythaemia rubra vera (PV), idiopathic myelofibrosis (MF), chronic lymphocytic leukaemia (CLL), hairy cell leukaemia (HCL) and diffuse large B-cell lymphoma (L). Morphometric evaluation was performed using a standard program set MicroImage (OLYMPUS). The greatest number of typical nucleated MK, “naked” nuclei, anucleated cytoplasmic fragments and the largest area were found in PV. The circular deviation factor of MK and their nuclei increased in all cases. The greatest number of clusters was observed in PV and HCL. A significant increase in the number of dysplastic and “naked” nuclei of MK was noted in all selected haematological diseases. The presence of neoplastic cells in bone marrow increased the morphological changes in MK. Quantitative and morphometrical significant differentiation of MK in separate microscopic field in the same slides confirms the necessity of performing trephine biopsies in each patient with haematological disorders.
Retinoic acid and transforming growth factor-ß (TGF-ß) affect differentiation, pro­liferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. The assay was based on [ 3H] thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in MCF-7 cells were also determined. Exogenous TGF-ß1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-ß 1 concentrations, a marked reduction in the stimulatory action of estradiol (10-9 and 10-8 M) was observed whereas in combination with tamoxifen (10 -7 and 10 -6 M) only 30 ng/ml TGF-ß 1 caused a statistically significant reduction to aproximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-ß 1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 ± 19% (con­trol = 100%) by 3 ng/ml TGF-ß1, and this dose was used throughout. It was found that addition of TGF-ß1 and isotretinoin to the culture did not decrease proliferation, while TGF-ß1 and tretinoin at low concentrations (3 x 10 -8 and 3 x 10-7 M) reduced the percentage of proliferating cells by aproximately 30% (67±8% and 67±5%, P < 0.05 compared to values in the tretinoin group). Both retinoids also led to a sta­tistically significant decrease in the stimulatory effect of 10-9 M estradiol, attenu­ated by TGF-ß1. In addition, the retinoids in combination with TGF-ß1 and tamoxifen (10–6 M) caused a further reduction in the percentage of proliferating cells. Immunocytochemical analysis showed that all the examined compounds gave a statistically significant reduction in the percentage of cells with a positive reaction to PCNA and Ki 67 antigen. TGF-ß1, isotretinoin and tretinoin added to the culture resulted in the lowest percentage of PCNA positive cells. However, the lowest fraction of Ki 67 positive cells was observed after addition of isotretinoin. The obtained results also confirm the fact that the well-known regulatory proteins Bcl-2 and p53 play an important role in the regulation of apoptosis in the MCF-7 cell line, with lowered Bcl-2 expression accompanying easier apoptotic induction. The majority of the examined compounds act via the p53 pathway although some bypass this important proapoptotic factor.
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