-
[show abstract]
[hide abstract]
ABSTRACT: Involvement of variety of genes, especially located on Y chromosome, is critical for the regulation of spermatogenesis. In particular, fertility candidate genes such as deleted in azoospermia (DAZ) are believed to have important function in sperm production, since DAZ is frequently deleted in azoospermic and severy oligozoospermic men. The role of the DAZ gene is supported by its exclusive expression in the testis and by its deletion in about 10% of azoospermic and severely oligozoospermic patients. The distribution of DAZ transcripts in seminiferous epithelium of human testis is reported in the present study. The use of Adobe Photoshop and Scion Image softwares allowed for semi-quantitative analysis of in situ RT-PCR (ISRT-PCR) results. The intensity of ISRT-PCR product's fluorescence was different within individual seminiferous tubules. It was clearly shown by using the pseudocolour scale and transforming the intensity of the fluorescence into levels of greyscale images. The more intense fluorescence characterised single spermatogonia and those organized in small groups inside separate tubules. The most intense accumulation of DAZ mRNA was observed in spermatogonia.
Folia Histochemica et Cytobiologica 02/2004; 42(2):119-21. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Ethane 1,2-dimethanesulphonate (EDS) causes apoptotic death of Leydig cells. Additionally, EDS causes damage of Chinese Hamster Ovary (CHO) cells but the occurrence of apoptosis is not such plentiful. The present study tested whether the inhibition of protein synthesis by cycloheximide (CHX) would influence apoptosis of CHO cells, induced by EDS. The study compounds induced morphological changes in CHO cell typical for apoptosis. An active form of caspase-9 and an alternation of mitochondrial transmembrane potential were also observed. In our study, a more cumulative effect of the CHX and EDS on apoptosis was observed, when both compounds were simultaneously employed. The obtained results indicated that synthesis of antiapoptotic proteins plays a very important role in the inhibition of apoptosis.
Roczniki Akademii Medycznej w Białymstoku (1995). 02/2004; 49 Suppl 1:11-3.
-
[show abstract]
[hide abstract]
ABSTRACT: Photodynamic therapy (PDT) represents a therapeutic approach in which photosensitised neoplastic cells undergo destruction under effect of light. In this study we have attempted to define effects of photochemotherapy on R2C cells, sensitised with protoporphyrin IX (PpIX) and to find out whether inhibition of gene expression by cycloheximide affects development of lesions in the cells. The photosensitised cells were exposed to visible light and development of apoptotic and necrotic lesions was followed in the cells, using the fluorescent staining with propidium iodide and Hoechst 33342. The experiments demonstrated that PpIX and light, acting in parallel, induce development of apoptotic and necrotic lesions in R2C cells. Intensity of the lesions correlated with concentration of the applied photosensitiser and with duration of light exposure. Using cycloheximide, we also inhibited protein expression in cells photosensitised with protoporphyrin before they were exposed to light. In the latter case, development of apoptosis was clearly intensified which might be explained by inhibition of anti-apoptotic protein synthesis in the cells.
Folia Histochemica et Cytobiologica 02/2002; 40(2):109-10. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The purpose of the study was to examine the phosphatidylserine translocation in human spermatozoa membrane during capacitation. Material consisted of human semen from normozoospermic men. Spermatozoa were stained with fluorescein-labelled annexin V. The presence and distribution of annexin V binding sites were analysed using the fluorescence microscope. Within first 60 min afterejaculation, 5-39% viable annexin V-positive spermatozoa were detected. The annexin V binding sites were found mainly in the midpiece. After 4 to 8 h of incubation of spermatozoa in capacitation medium (BMI), the number of cells positively stained with annexin V increased. After capacitation, the localisations of phosphatidylserine was changed and the annexin V binding sites were found also in the acrosomal region but never in the equatorial area. The process of the phosphatidylserine translocation observed during our experiments may reflect changes of the plasma membrane occurring during capacitation or, less likely, apoptosis of spermatozoa.
Folia Histochemica et Cytobiologica 02/2002; 40(2):111-2. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: In women with CIN at fertile age and those over 50 years of age, EGFR expression is lower in the presence of Chlamydia trachomatis (Cht) infection. In all Cht infected women over 50 years of age expression of Ki 67 is higher; the increase is significant among women with invasive carcinoma. In these groups of women with CIN and invasive carcinoma TGF-alpha expression is insignificantly augmented. Chronic Cht infection is associated with cervical hypertrophy.
European journal of gynaecological oncology 02/2002; 23(6):511-3. · 0.47 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Human papillomavirus type 16 (HPV16) is a major agent in cervical cancer etiology. Its early proteins are responsible for virus persistence, replication and initiation of neoplastic disease. In the present study we describe a use of baculovirus-insect cell expression system for production and study of HPV16 E2 and E4 proteins. The E2 protein binds specifically to viral DNA and E4 protein shows characteristic cytopathic effects on cells.
Folia Histochemica et Cytobiologica 02/2001; 39(2):125-6. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Lectin from Chelidonium majus L. (CML) significantly stimulates the proliferation of human lymphocytes and has hemagglutination activity towards group B human erythrocytes and potent antimicrobial properties against multiresistant enterococci and staphylococci. In the present work we describe the effect of lectin from Chelidonium majus L on normal and cancercells in culture in vitro. The studies were performed on three types of cells: CHO, R2C and on normal mouse fibroblasts. Effects on the cultures were examined 24 h after addition of CML. Exposure to CML resulted in growth inhibition of CHO and R2C cells but not of fibroblasts. Moreover, evident apoptotic lesions were observed in CHO cells and less well marked apoptotic lesions in R2C cells. In contrast, only insignificant numbers of fibroblasts reacted to the applied lectin.
Folia Histochemica et Cytobiologica 02/2001; 39(2):215-6. · 0.81 Impact Factor
-
Advances in experimental medicine and biology 02/2001; 495:435-8. · 1.09 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Normal course of spermatogenesis reflects appropriate expression of specific genes in spermatogenic cells. In the present study, the localization of DAZ mRNA in human testis obtained from two fertile organ donors has been determined. It was established that DAZ mRNA is located in the germinal epithelium of seminiferous tubules. The most intensive accumulation of DAZ transcript was observed in primary spermatocytes. The results indicate that expression of DAZ gene begins in spermatogonia and that distribution of DAZ mRNA most likely correlates with the stage of the human seminiferous cycle.
Folia Histochemica et Cytobiologica 02/2001; 39(2):117-8. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Photodynamic therapy (PDT) represents a therapeutic approach in which photosensitised neoplastic cells undergo destruction under the effect of light. In this study we have attempted to define effects of PDT on CHO cells, sensitised with protoporphyrin IX (PpIX). The photosensitised CHO cells were exposed to a visible light and development of apoptotic and necrotic lesions was followed in the cells, using the fluorescent staining with propidium iodide and Hoechst 33342. The experiments demonstrated that PpIX and light, acting in parallel, induce development of apoptotic and necrotic lesions in the cells. Intensity of the lesions was correlated with the concentration of the applied photosensibiliser and with the duration of exposure to light. The control experiments suggest that development of apoptosis in the applied model probably reflect mitochondrial damage, while processes developing close to the cell membrane are responsible for necrosis. In order to corroborate the obtained results, ultrastructural studies were performed on experimental groups in which evident apoptotic lesions were observed in the cells.
Folia Histochemica et Cytobiologica 02/2001; 39(2):213-4. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The aim of the study was to assess the expression of progesterone membrane receptors in human spermatozoa before and after capacitation. The sperm of 16 men with normozoospermia and 48 men with oligozoospermia was examined. Progesterone-BSA complex labelled with fluorescein isothiocyanate (P-BSA-FITC) was applied to visualise the progesterone receptors. The spermatozoa were capacitated with BM1 medium. In freshly ejaculated sperm from normozoospermic men, P-BSA-FITC staining (bright fluorescence of acrosome region) was observed in 50.1+/-5.1% of spermatozoa. Following capacitation in BM1 medium, the percentage of P-BSA-FITC stained spermatozoa increased to 69.9+/-3.3%. In sperm from slight oligozoospermia the percentage of P-BSA-FITC stained cells before and after capacitation was 48.2+/-8.5% and 67.9+/-4.2%, respectively. In men with severe oligozoospermia the percentage of P-BSA-FITC stained cells was 11.9+/-1.7% and 11.9+/-1.9%, respectively. It is supposed that progesterone membrane receptors in human spermatozoa are gradually exposed during capacitation. Disturbances in the expression of progesterone membrane receptors might be involved in male infertility.
Folia Histochemica et Cytobiologica 02/2001; 39(2):139-40. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The presence of aneuploidy in spermatozoa influences their biological characteristics, especially their ability to fertilise the ovum. The aim of the present study was to investigate if aneuploidy is accompanied by any changes in the morphology of spermatozoa in oligozoospermic patients. For this purpose, the percentage of aneuploid cells in sperm and the correlation between the specific morphological forms of spermatozoa and aneuploidy were evaluated. The study proved a negative correlation between DNA content of aneuploid and normal spermatozoa. A weak positive correlation was demonstrated between the presence of aneuploid spermatozoa and DNA content of spermatozoa with large heads. No such correlations could be detected for DNA content of the remaining morphological forms of spermatozoa. Thus, men with a lowered number of spermatozoa and/or with abnormal spermatozoal morphology should have their spermatozoal DNA content tested in order to evaluate the degree of aneuploidy, especially in cases where in vitro fertilisation is intended.
Folia Histochemica et Cytobiologica 02/2001; 39(1):43-6. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: In 28 Spitz nevi DNA content was determined by video-imaging cytometry. The nevi were selected for this study because of difficulties in differentiation from melanoma and heterogeneity of this type of nevus. Morphological features of Spitz nevi and differences helpful for differentiation between Spitz nevi and malignant melanoma were identified. DNA ploidy was measured in paraffin embedded and fresh tissue material from each patient and the results were comparable. The sample preparation process and video-imaging method are presented in this study. Twenty two (78.6%) lesions contained diploid cell populations, 5 (17.9%) aneuploid and 1 (3.6%) tetraploid cell population. A significant correlation was observed between DNA ploidy measured in fresh tissue and retrospective material. The results indicate the presence of abnormal DNA content in some of the lesions. This observation does not indicate that DNA ploidy cytometry is useful for the differentiation of Spitz nevi from malignant melanoma.
Polish journal of pathology: official journal of the Polish Society of Pathologists 02/2000; 51(1):45-50. · 0.35 Impact Factor
-
Folia Histochemica et Cytobiologica 02/1999; 37(2):123-4. · 0.81 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The goal of the study was to evaluate ultrastructural changes in aneuploid human spermatozoa. The sperm was collected from seven males with teratozoospermia. In all patients ultrastructural changes in spermatozoa were noted: inappropriate nuclear chromatin condensation, changes in the formation of the acrosome and pathology of the mobile apparatus. These changes suggest a defect in spermatogenesis at the level of spermatide or spermatocyte. Ultrastructural spermatozoa studies may reveal new elements related to factors responsible for decreased sperm fertilizing ability.
Ginekologia polska 07/1998; 69(6):401-4. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A genetic etiology has been proposed for severe forms of idiopathic male infertility and involvement of variety of genes is critical for the regulation of spermatogenesis. The role of DAZ gene is supported by its exclusive expression in the testis and deletions of DNA sequence within this gene are associated with azoospermia or severe oligozoospermia. The objective of our study was to validate the PCR strategy, using primers based on cDNA sequence, for the detection of genomic aberration among males with azoospermia or severe oligozoospermia who carry mutations in the DAZ gene. Using the PCR technology, deletion of the DAZ gene in one infertile male with azoospermia was detected. No deletion was detected in any of the remaining 15 patients.
Ginekologia polska 07/1998; 69(6):420-4. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: In the studies, changes in concentration of free calcium ions were analysed in human spermatozoa, as affected by steroid hormones. The experiments were performed on human sperm preparations originating from healthy donors which fulfilled criteria of WHO (1992). The cells separated by the swim-up technique were loaded with the Fura-2AM (4 mM). The changes in concentration of Ca2+ were monitored employing the ratio-imaging technique (MagiCal, Applied Imaging, England). We examined the influence of progesterone, hydrocortisone, RU 486, testosterone, 17 b-estradiol, pro-BSA on Ca2+ concentration. The performed experiments demonstrated that already a few seconds (5-150s) exposure to progesterone and prog-BSA induced a dose dependent increase in Ca2+ concentration in normal spermatozoa. The cell characteristics (no transcription), reaction time, dose dependent and the reaction to pro-BSA, show that progesterone acts through the membrane receptors.
Ginekologia polska 07/1998; 69(6):430-6. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The testis is divided, both functionally and anatomically, into two specialised compartments, i.e. the vascular interstitial tissue and avascular seminiferous tubules. Leydig cell steroidogenesis is mainly controlled by luteinizing hormone (LH) secreted from the anterior pituitary. It is well known that LH is able to generate second messengers by adenylate cyclase and the cycle of inositolphospholipids. Also Ca2+ ions are taken into account as an important modulators of Leydig cell steroidogenesis. The aim of this study was to show the effect of macrophage-conditioned medium on the basal and LH stimulated testosterone secretion by mouse Leydig cells in vitro. A source of Leydig cells was the testis of mature, Swiss strain mice. Leydig cells were isolated and cultured for 48 hrs. Testosterone levels were measured radioimmunologically. Intracellular free Ca2+ ions were analysed by computer automatic system 'MAGICAL'. Results indicated that not only LH but also macrophage-conditioned medium were able to modulate testosterone secretion by Leydig cells and increase intracellular calcium concentration. Therefore, it seems possible that the action of proteins secreted from macrophages is mediated through Ca2+ ions which are involved in another signal transduction pathway leading to stimulation of testosterone biosynthesis by Leydig cells in vitro.
Ginekologia polska 07/1998; 69(6):441-5. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The aim of our study was to analysis the ploidy of spermatozoa from oligozoospermic individuals. The study was performed in 48 patients with oligozoospermia. Spermatozoal DNA was analysed using MagiCal imaging cytometry. The percentage of aneuploid cells was determined using CYT-2 program. The results indicated aneuploidy in 73% of the examined patients. The amount of cells with abnormal DNA content ranged from 30 to 100%. Monitoring of the DNA in human spermatozoa may have practical applications in establishing the reasons for infertility as well as for screening of the semen for assisted reproduction techniques.
Ginekologia polska 07/1998; 69(6):517-23. · 0.41 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The fully automatic method for reconstruction of spermatozoa trajectories was presented. The method is based on computer image analysis. It allow to process pure semen without any treatment like diluting or purifying. Since no human interacting is available, this method lead to objective results. The method was used to analyse semen taken from healthy men (WHO 92 standard). The result shows absence of different populations in single sample.
Ginekologia polska 07/1998; 69(6):532-6. · 0.41 Impact Factor
