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Late onset hypogonadism is a common condition in aging males. The population of men who can be potentially treated with testosterone supplementation is growing. Controversy that surrounds testosterone replacement therapy is due to generally undefined lower limits of normal testosterone level and high prevalence of hypogonadal symptoms in elderly population and the non-specific nature of these symptoms. Incidence of prostate hyperplasia and occult prostate cancer in elderly are both high. The risk that testosterone treatment would trigger prostate cancer was not fully recognized. The aim of this mini review is to present a risk of carcinogenesis within the prostate related to testosterone treatment.

Abstract: Ciprofloxacin is a chinolone antibiotic, which is used mainly in the treatment of urinary tract infections but also in pulmonary tract, prostate gland, bone and bone marrow infection. Ciprofloxacin is also known for its anticancer in vitro properties. In this study hamster ovarian cancer line CHO AA8 was used for evaluation of cytotoxic properties of ciprofloxacin against neoplastic cells. For this purpose we used different concentrations of ciprofloxacin range from 10 to 1000 microg/mL. Cell viability was counted using trypan blue assay. Ciprofloxacin induced morphological changes and decreased viability in a concentration and time dependent manner within CHO AA8 cells. In low concentrations cytotoxic effect of ciprofloxacin is weak only after 24 h incubation. In the highest concentration of ciprofloxacin, after 24, 48 and 72 h incubation only a very small number of living cells (not exceeding 1%) was observed. No living cells were observed after 96 h of incubation times and ciprofloxacin concentrations of 800 and 1000 micrpg/mL. These promising results deserved future studies on chinolones and ovarian cancer.

Objective: The aim of this study is to report our experience with laparoscopic radical cystectomy (LRC), evaluating the technique and perioperative and pathological outcomes., Methods: 47 LRCs were performed due to muscle-invasive bladder cancer. Conduits were performed in 23 patients and neobladders in 23 (one bi-intestinal). One ureterocutaneostomy was created., Results: In 43 patients LRC was performed with minilaparotomy for urinary diversion. The mean operation time was 290 min. Four operations were converted. Complications included sigmoid colon injury, urinary leak, lymphatic leak, short-term paralytic ileus, and heart attack. Mean blood loss was 220 ml. Hospitalization time was 6 days. Tumor stage was pT2b, pT3a, pT3b, and pT4a in 28, 13, 5, and 1 patient, respectively. No positive margins were found. The mean number of lymph nodules was 17, while in the last 25 procedures it was 21. 17% of patients had tumor in the lymph nodes. The mean follow-up was 10 months. Local recurrence and dissemination was observed in 2%. Continence in patients receiving neobladder was fully satisfactory., Conclusions: More complications are related to neobladder than to ileac conduit. LRC with minilaparotomy seems to be an attractive treatment option for patients with muscle-invasive bladder cancer. Radical cystectomy performed intracorporeally could be reserved for 'robot-assisted' operations., (Copyright © 2010 S. Karger AG, Basel.)

Cultured human melanocytes are increasingly being used in the treatment of vitiligo. The growth media contain various types of mitogenic factors, both recombinant human (e.g., rhbFGF and rhSCF) and synthetic (e.g., TPA). High concentrations of mitogenic factors accelerate the cell cycle, and consequently may increase the risk of carcinogenesis of transplanted cells. Mutations of genes of the RAS/RAF/MEK/ERK signaling pathway are very often found in the early stages of the development of melanoma. TPA is considered to be an oncogenic factor, but so far there is no evidence to show that it is responsible for damage to the genetic material of cultured melanocytes. The aim of our study was to assess the risk of the development of mutations in selected genes of the RAS/RAF/MEK/ERK signaling pathway during the culturing of melanocytes in various growth media. Based on the results obtained, it can be concluded that TPA and high concentrations of other growth factors intensify the proliferation of melanocytes, without the risk of damage to the HRAS (exon 1 and 2), KRAS (exon 1 and 2), NRAS (exon 1 and 2), and BRAF (exon 11 and 15) genes. In order to assess the total safety of the transplantation of cultured melanocytes, it is necessary to carry out further studies on other signaling pathways as well as carry out biological tests on an animal model.

Background: A variety of tissue engineering techniques are currently under development or investigation for bladder augmentation, but so far no approach is clearly superior. The aim of this study was to compare the suitability for cystoplasty augmentation in rats of in vivo implanted acellular bladder matrices (BAM) previously seeded with hair follicle stem cells and that of matrices implanted without the cells., Materials and Methods: The rat hair follicle stem cell line was positive for CD34, p63, and Ki-67. 1 x 10(6) cells from 34 to 40 passages seeded onto nine BAM scaffolds were cultured for one week. Nine other scaffolds were left unseeded. Scaffolds were grafted into a surgically created defect within the anterior bladder wall: nine rats with acellular matrices and nine with cell-seeded BAM. Rats observed for six months were killed in monthly intervals. We performed gross examination, X-ray cystography, and hematoxylin-eosin, cytokine (CK)-7, CK-20, myoglobin, and desmin staining of the excised bladders., Results: Minimal adhesions were observed and urinary leakage was noted in one case. Two animals died in the acellular group. Rats developed stone disease in bladders reconstructed with acellular BAM. Bladder capacity was similar, but the shape was regular and characteristically oval only in bladders grafted with cell-seeded BAM. Muscle layers in the apical parts of the reconstructed bladder walls were extremely thin in the cases of acellular grafts and thicker in bladders reconstructed with cell-seeded grafts. Muscle layer regeneration was better in the cell-seeded group. Urothelium regenerated in all animals., Conclusions: We have shown that hair follicle stem cells may be used for rat bladder wall regeneration.

Background: In vitro-constructed grafts can be used for human bladder augmentation. There are many diseases in which autologous cells cannot be used for this purpose. The aim of the present study was to examine the potential of rat vibrissae hair follicle cells to form cultures, which could serve as a source for in vitro creation of urinary bladder wall grafts., Methods: Two hundred vibrissae were excised from young Wistar male rats. Two different digestions were performed, in dispase and in collagenase. All follicles were additionally incubated in trypsin and ethylenediamine tetraacetic acid. Two different culture media based on DMEM (Dulbecco's Modified Eagle's Medium) were used: the first was supplemented with keratinocyte growth factor (KGF) and the second with epidermal growth factor. Immunocytochemical detection of cytokeratin, CD34, p63, Ki-67 (proliferation index), and HMB45 (Human Melanoma Black 45) was performed., Results: Forty-eight primary cultures of rat follicle vibrissae cells were established from 200 hair follicles (24% successful rate). Twenty-four primary cultures were obtained after dispase digestion and 24 after collagenase treatment. Each group was cultured in 2 different media. A heterogeneity of primary cultures was observed. Cells formed a monolayer within a period of 2 to 4 weeks. The 24 primary cultures established after dispase treatment exhibited monolayers of small cuboid cells expressing cytokeratin and CD34. In the 40th passage 20%-40% of cells expressed p63; 85% of these cells from late passages were positive for Ki-67, indicating preserved mitotic potential. Epithelial-like phenotype was observed after dispase digestion and cultivation in KGF-supplemented medium. After 3 weeks, the morphology of these cells changed into fibroblast-like. These cultures were negative for CD34. Fibroblast-like cell growth was observed after collagenase treatment in both KGF- and EGF-supplemented media. These cells were positive for the melanocyte cell marker (HMB45)., Conclusions: Culture media and isolation conditions influence hair follicle stem cell differentiation. The stem cell niche within the hair follicles is a reservoir of cells, which can be potentially used for in vitro creation of urinary bladder wall grafts.

Unlabelled: Several platinum(IV) complexes are showing considerable promise in initial trials, producing reactive intermediates that then interact with DNA., Aim: To perform in vitro study of two new platinum(IV) complexes cytotoxic effect on B16 mouse melanoma cells., Methods: PtCl(4)(dbtp)(2) and PtCl(2)(6mp)(2) complexes were prepared. PtCl(4)(dbtp)(2) was created as modification of PtCl(4)(dmtp) test previously. Apoptosis and necrosis were examined using flow cytometry, upon Annexin V/PI staining., Results: LC(10), LC(50) and LC(90) parameters established for PtCl(4)(dbtp)(2) were as following: 2.6, 17.0, 58.0 mumol/L. However LC(10) and LC(50) established for PtCl(2)(6mp)(2) were 1.2 and 14.0 micromol/l respectively. The both complexes induced apoptosis. PtCl(2)(6mp)(2) induced cell cycle arrest in G0/G1, while PtCl(4)(dbtp)(2) - in S-phase., Conclusions: PtCl(4)(dbtp)(2) appeared to be more cytotoxic against B16 cells than PtCl(2)(6mp)(2). Apoptosis was the main mechanism of cell loss in cultures incubated with both tested complexes.

There are many diseases in which autologous urothelial and muscle cells cannot be used for in vitro construction of the urinary bladder wall for augmentation (cystoplasty). These diseases are the most frequent indications for bladder augmentation. The present paper focuses on the idea of harvesting potentially multipotent stem cells out of hair follicles in order to use them for regeneration of the urinary bladder wall. Current clinical practice suggests the use of cultures enriched with progenitors. The hair follicle stem cell niche gives an opportunity to reduce the invasiveness of harvesting these cells. Both epithelial and dermal multipotent stem cells populations within hair follicles raise new possibilities for tissue engineering of the urinary bladder. The hypothesis is that hair-follicle stem cells can be used, with the guarantee of the sufficient cell number, for a construction in vitro of the urinary bladder wall replacement.

No survival profit has been achieved for patients with disseminated prostate cancer since hormonal therapy was introduced. It is proposed that dissemination of rare prostatic cancer stem cells may lead to metastatic disease and that resistance of these cells to androgen ablation makes them responsible for failure of current treatments. In this paper, we will discuss the significance of the stem cell model for understanding prostate cancer pathogenesis. The concept of prostate cancer as a stem cell disease has the potential to change our view of its treatment in the particular case of disseminated disease. The major cellular target of prostate cancer therapy has to be directed against neoplastic stem cells. The combination of molecular-targeted therapy with the concept of the cancer stem cells should be introduced for the treatment of disseminated prostate cancer. Disseminated prostate cancer must be treated with agents directed toward stem cells, while hormone-therapy must be only an additional treatment leading to the decrease of tumor burden.

Unlabelled: Pancreatic islet transplantation is a treatment method for type I diabetes. Its outcome is influenced by numerous factors, islet quantity and function being important ones of them., The Aim of This Study: was to estimate the influence of pancreas preparation method on the outcome of islet isolation in rat., Material and Methods: 6 pancreata harvested from Lewis rats were used in this research. Pancreatic duct was cannulated and pancreas was injected with 1 mg/ml collagenase P solution (Sigma) and then excised. After cutting into smaller fragments, it was digested in collagenase P solution for 15-20 min. Enzyme activity was then stopped by adding dilution medium. Heterogenous cell suspension was centrifuged in density gradient (Gradisol) to isolate islets. Pancreatic islets were collected and islet equivalent was calculated. Islet purity degree was estimated as islet cells to all cells, including exocrine, ratio. Islet viability was estimated using propidium iodide and fluorescein diacetate staining. Photographic documentation was made., Results: Proper islet morphology, highest number and viability was obtained when pancreas was excised properly (isolation 3 and 4)., Conclusion: Pancreas preparation method is one of which influences on islet isolation outcome.

It was shown that ointments containing silver compounds delay wound healing and increase the risk of hypertrophic scarring in burned patients. The aim of the study was to establish the influence of AgNO3 on cell viability and apoptosis of keratinocytes and fibroblasts in vitro. Foreskin was used to establish primary human keratinocyte and fibroblasts cell cultures. Keratynocytes grew in DMEM/Ham's-12 at 3:1 ratio, supplemented with 10% FBS, EGF, insulin, transferin, triiodothyronine and hydrocortisone. Fibroblasts were cultured in DMEM with addition of 10% FBS. The influence of AgNO3 on keratinocyte and fibroblast cultures was evaluated by fluorescence microscopy and flow cytometry. Double staining with Annexin V-FITC and propidium iodide was performed. The AgNO3 at lower concentration (3 and 15 x 10(-4) M/dm3) than used for patient's treatment (31 x 10(-4) M/dm3) revealed to be toxic and trigger apoptosis in human keratinocytes and fibroblasts. The ointments containing nitrates should be used with caution especially in conditions where the epithelial layer is destroyed. The nitrate can negatively affect wound healing.

The concept of cancer as a stem cell disease has the potential to dramatically change our view of the problem of its treatment. A major cellular target of cancer therapy will be directed against neoplastic stem cells. Current investigations are attempting to combine molecular targeted therapy concepts with the concept of the cancer stem cells. Molecular targeted therapy against cancer stem cells can be divided into 6 categories that correspond to following types of targets, presented in this review: surface molecules; ABC multidrug resistance proteins; specific oncoproteins; normal and neoplastic stem cell pathways; survival factors, present in cancer stem cells, such as NF-kappaB pathway, PI3K, downstream signaling molecules and related signal-transduction pathways and oxidative stress.

Aim: To analyze cell viability and morphology of primary cell cultures from CD133 immunolabeled and sorted cells from epithelium of patients suffering from benign prostate hyperplasia (BPH)., Methods: Cells obtained from 5 patients were divided in two fractions. First fraction (CD133+/CD133-) was cultivated in DMEM with 10% FBS. Second fraction was mixed with CD133 microbeads and immunomagnetically divided into CD133+ and CD133- fractions. These cells were cultivated and followed-up for 2 weeks. Cells were stained for Annexin V FITC/propidium iodide., Results: Seventy CD133+/CD133- cultures, thirty-one of CD133+ and thirty-one of CD133- cells were established. There were 5-fold and 3-fold increase of CD133+/CD133- and CD133+ cell number after 2 weeks, respectively. CD133+/CD133- and CD133+ monolayers displayed epithelial-like morphology and cytokeratine expression. CD133- cultures collapsed. Cell viability within CD133+ and CD133- populations was 90.1-/+6.3% and 24.3-/+6.2%, respectively. Apoptotic index was 9.0-/+6.1% and 28.5-/+23.8% within CD133+ and CD133- cultures, respectively., Conclusions: CD133 separated human primary epithelial cell cultures displayed differences in morphology, viability and apoptosis occurrence. Immunomagnetic sorting can be recommended in each in vitro experiments with primary cell cultures in order to provide more objective results.

Introduction: Alginate scaffolds are widely used in tissue engineering. The aim of this study was to evaluate alginate as a scaffold for 3D cultures of rapidly proliferating cells., Materials and Methods: Murine 3T3 fibroblasts were cultured in an alginate scaffold for 30 days. Cells growing in alginate were observed under the inverted microscope. Pathologic examination by hematoxylin and eosin staining was done at the end of the experiment., Results: Migration of rapidly proliferating cells from the 3D scaffold and an inappropriate growth pattern were observed during the experiment. Cells and scaffold did not form a solid graft., Conclusions: The results obtained in this study indicated that alginate is not a good biomaterial for a durable implant.

Background: An interesting way to regenerate a kidney is an autologous bone marrow transplantation. The aim of this study was to examine whether chronic kidney disease influenced bone marrow progenitors., Methods: Wistar male rats included group I (n = 4, chronic kidney disease 1/2, CKD 1/2) that underwent right nephrectomy. In group II (n = 3, chronic kidney disease 5/6, CKD 5/6) underwent removal of the right kidney and approximately one-third of the cortex of the left kidney. Animals in the control group (n = 4) were intact. Bone marrow cells obtained from femurs were separated using a CD34 Micro-Beads magnetic isolation kit. Isolated cells were counted using a trypan blue exclusion test. Numbers of isolated cells were presented as mean values with standard deviation with P < .05 considered significant. CD34(-) cells were cultivated and observed to the passage 6., Results: The CKD rat model was used for in vitro experiments. There were no differences in cell numbers isolated from control rats versus both CKD rats. No differences were observed in CD34(-) cells after separation when compared to controls. Cell morphology was similar in primary CD34(-) cultures during the first days of primary culture. CD34(-) primary cultures established from chronic renal failure rats collapsed within 2 weeks. No differences were found in CD34(+) cell number after isolation when compared with controls. These cells did not form a monolayer. Cells in cultures established from control animals resembled normal fibroblast-like morphology of mesenchymal stem cells during 3 months., Conclusions: Bone marrow cells from chronic renal failure rats showed no capacity for in vitro proliferation. We speculated that bone marrow cells obtained from renal chronic failure patients may not be useful for autologous cell transplantation.

Introduction: The aim of this study was to evaluate polyasparagine (PAA), a new, promising scaffold. PAA was compared with alginate, which is used in cell transplantations and may be regarded as a standard., Materials and Methods: In vitro cell viability on both scaffolds was assessed. C57B1 mice were injected s. c. alginate or PAA with or without 3T3 cells. After two months specimens from sites of injection were examined and blood samples were taken for enzymatic activity estimation. Cathepsin D activity and alpha1-antitrypsin levels were measured., Results: In vitro cell viability was lowest on PAA and highest in control group. Increase in levels of enzymes measured was observed in response to PAA and alginate and was lower in case of polymer seeded with cells. Increase in alpha1-antitrypsin levels was lower in case of PAA in comparison to alginate. Scaffold degradation in histopathological specimens was visible., Conclusion: The results indicate that PAA implants undergo biodegradation and nonspecific inflammatory response is comparable to alginate.

Unlabelled: To develop a tissue-engineered bladder wall replacement with elements obtained from non-urinary tract components is an atractive idea. The aim of this study was to compare growth of hair follicles epithelial stem cells and mesenchymal stem cells to urothelial cells and fibroblasts cells on scaffold prepared from rat collagen type I., Materials and Methods: Wistar rats were used in experiment. Rat urothelial cells, hair follicles epithelial stem cells, mesenchymal stem cells and 3T3 cells were cultivated in DMEM (Sigma) supplemented with 10% (or 20% for hair follicles cells) of Fetal Bovine Serum (FBS). Epithelial cell cultures were suplemented with EGF (10 ng/ml; Sigma). Cells were stained using anti-cytokeratine (Clone MMF) and anti-cytokeratine 7. Anti-CD34 and anti-p63 staining were done. Collagen scaffold was prepared from tendoms of Wistar rat's tails. 6-well plates were covered with collagen scaffold. 25 x 10(3) of cells were seeded on each well and cultured for a week. Cells in the controls were seeded on polystyrene surface. After a week cell viability was assessed using MTT test (Sigma). Each experiment was triplicated. Photo documentation was prepared. The differences between means were compared using t-Student test., Results: There were 106.5 +/- 23.4 x 10(3) and 310.7 +/- 60.7 x 10(3) of 3T3 fibroblasts growing on polystyrene and collagen, respectively (p < 0.05). The initial cell number was 25.0 x 10(3). Urothelial cells expressed epithelial markers. There were 40.0 +/- 4.2 x 10(3) and 4.5 +/- 1.8 x 10(3) urothelial cells growing on polystyrene and collagen, respectively after 7 days of culture (p < 0.01). There were 118.5 +/- 19.7 x 10(3) and 114.1 +/- 33.2 x 0(3) of mesenchymal stem cells growing on polystyrene and collagen, respectively (NS). Hair follicles epithelial cells expressed epithelial markers and were slightly positive for CD34 and p63. There were 292.5 +/- 33.3 x 10(3) and 167.4 +/- 24.9 x 10(3) of hair follicles epithelial cells growing on polystyrene and collagen, respectively (p < 0.05). Collagen scaffold decreased proliferation of follicle epithelial and urothelial cells., Conclusions: Hair follicles epithelial stem cells and mesenchymal stem cells can be potentially used in tissue-engineering, with the guarantee of the sufficient cell number for transplantation. It seems that construction in vitro of urinary bladder walls from elements obtained from non-urinary tract tissues is feasible.

Background: Chronic alcohol consumption induces an increase in oxidative stress. There are studies that indicate changes in the activity of antioxidant enzymes and the concentration of lipid peroxidation products in the blood of organisms as a result of alcohol consumption, but published results are somewhat conflicting. The aim was to study the effect of alcohol withdrawal and detoxification on the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in erythrocytes and on the concentration of thiobarbituric acid-reactive substances (TBARS) in the erythrocytes and blood plasma of alcohol-dependent patients., Material/methods: The study group consisted of 42 alcohol-dependent men who underwent seven-day detoxification at an addiction treatment department. The control group consisted of 20 healthy male volunteers. Blood for analysis was taken from the antecubital vein before detoxification, after the first detoxification, and after several (> or =3) detoxifications., Results: Before detoxification, the concentration of TBARS in the men's blood plasma was 40% higher (p<0.001) than in the control group. After detoxification, the concentration of TBARS decreased, but remained higher than in the control group. The activities of SOD and GPx in erythrocytes of alcohol-dependent persons were statistically significantly lower than in the control group both before and after detoxification. However, after detoxification, a slight increase in the activity of SOD was observed compared to that before detoxification., Conclusions: This study confirms that alcohol-dependency leads to oxidative stress in the peripheral blood. Seven-day alcohol withdrawal and detoxification has no effect on TBARS concentration and antioxidant enzyme activity.

Purpose of Investigation: The aim of the study was to determine the activity of cathepsin D (CTSD) and alpha-1-antitrypsin (AAT) in the blood serum of women with cervical carcinoma treated with different modes of therapy., Methods: The study was conducted on 68 women suffering from carcinoma of the uterine cervix, that were irradiated intracavitarily by a Selectron LDR brachytherapy unit. Additionally, all patients were treated with different therapy methods according to clinical stage., Results: In women with cervical cancer, CTSD activity was higher while AAT activity was lower both before and after brachytherapy sessions as compared to controls. Six months after the end of therapy, the activity of CTSD and AAT reverted back to the values characteristic for healthy women., Conclusion: The estimation of cathepsin D and alpha-1-antitrypsin activity during the course of cervical cancer management may be useful in early detection of potential recurrence and/or widespread metastasis formation.

Abdominal hernia repair is one of the most common surgical procedures. Current data indicate that the best treatment results are achieved with use of synthetic material to reinforce weakened abdominal wall. Prosthetic materials utilized for hernia repair induce adhesions with underlying viscera. They should be therefore separated from them by a layer of peritoneum otherwise adhesions may cause to serious complications such as bowel-skin fistulas. The aim of our work was to determine if implantation of our collagen membrane into abdominal wall defect induce adhesions in rat model of ventral hernia. The collagen film was obtained by acetic acid extraction of rat tail tendons and than casting the soluble fraction onto polyethylene shits. Abdominal wall defect was created in 10 Wistar male rats. Collagen membranes were implanted into the defect using interrupted polypropylene stitches. After 3 months of observation all animals were sacrificed. No adhesions between path structure and bowel developed. In one often rats (10%) adhesion between fixating stitch and omentum was observed. Complete mesothelium lining and vascular ingrowth were microscopically observed within implanted structure. Promising result requires further confirmation in a larger series of animals.

Introduction: The function of reconstructed bladder depends on the contracting bladder wall. This can be obtained with a proper innervating segment. Polyglycolic acid (PGA) is used as cell vehicle. Chitosan supported the adhesion and differentiation of neurons. The aim of the study was to compare PGA with PGA/chitosan 'sandwich' grafts for bladder regeneration., Methods: 3T3 fibroblasts were seeded on 6 PGA and on 3 chitosan scaffolds and incubated for 3 days at 37 degrees C in 5% CO(2) before implantation. Three rats underwent bladder reconstruction with PGA cell-seeded grafts and 3 with PGA grafts covered with chitosan cell-seeded grafts ('sandwich' graft). Three rats in the control group were not operated. After 6 months, reconstructed tissue was stained with hematoxylin and eosin. Neurons were identified by synaptophysin and neuron-specific enolase staining., Results: No complications were noticed. PGA/chitosan grafts were evaluated as (+++) and (++), while PGA grafts were evaluated as (++) and (+) with use of synaptophysin antibody. The control group was evaluated as (+). PGA/chitosan grafts were evaluated as (++) and (+), while PGA grafts were evaluated as (++) and (+) in neuron-specific enolase staining. The control group was evaluated as (+)., Conclusion: Chitosan improved PGA's abilities as a cell matrix and in guiding neurons into the graft., (2008 S. Karger AG, Basel)

Doxazosin triggers apoptosis via an imprecisely defined receptor mechanism that is related to tumor necrosis factor receptors (TNFRs). The aim of this study was to determine CD95, TNFR-1, TNFR-2, CD40 expression in primary prostate epithelial cultures incubated with doxazosin. Epithelial cultures were cultivated from 10 benign prostate hyperplasia patients. The cells were incubated with 20, 50 and 80 microM of doxazosin. Apoptosis was confirmed by fluorescence microscopy and flow cytometry. The cells were analyzed for expression of FAS, CD40, TNFR-1 and TNFR-2 by flow cytometry. Early apoptotic cells were present in all groups. A positive correlation was noticed between doxazosin dose and TNFR-1-, -2-positive cells. A decrease of CD40-positive cell population was observed in the lowest concentration. A decrease of mean fluorescence intensity signal of CD40 and CD95 was observed in the lowest concentration. Doxazosin-triggered apoptosis was dose-independent. The initiation of apoptosis was a result of receptors 'crosstalk' rather than a single receptor pathway activation. TNF receptor self-assembly process should be checked as a potential mechanism leading to apoptosis after doxazosin treatment.

Tissue engineered blood vessels are an attractive alternative for synthetic vascular prostheses utilized in vascular reconstructions. Continued for over 20 years in vitro and animal experiments has recently gave an opportunity to utilize this technique in clinical medicine. Satisfying results achieved with this approach in humans encourage wider use of vascular bioprosthesis in the future.

Hepatocellular carcinoma and glioblastoma are both very aggressive forms of human neoplasms. The most effective treatment includes the combination of surgery, chemotherapy and radiation. Sodium butyrate (NaB) demonstrates a high efficiency and low toxicity. It inhibits proliferation and cell cycle of the cancer cells. The aim of this study was to investigate the effect of sodium butyrate on HepG2 and C6 cell line viability. Hepatocellular cancer (HepG2) and glioblastoma cell line (C6) were cultured in DMEM/Ham's F12 medium with 10% FBS and antibiotics. Different NaB concentrations (0-10 mM) were tested. The control consisted of cells without tested substance. The incubation times were 24 and 48 h. Cell viability was studied using Trypan Blue exclusion test. Inhibitory influence of sodium butyrate on cell viability in both examined cell lines was confirmed. Strong correlation between NaB concentration and cell viability after 24 h was noticed (correlation coefficient was 0.94 and 0.98 for C6 and HepG2, respectively). IC50 values after 24 h were 8.44 mM and 6.17 mM for C6 and HepG2, respectively. The strongest effect was observed after 48 h of incubation with NaB. IC50 values were 3.44 mM and 1.47 mM for C6 and HepG2 (correlation coefficients after 48 h were 0.91 and 0.631 for C6 and HepG2, respectively). C6 line was more resistant to NaB than HepG2. Both cell lines were sensitive to NaB treatment, which gives the promise that NaB can be used against broader spectrum of neoplasms in the future.

It was a chance discovery that ambroxol parenteral administration led to urinary bladder stone formation in rats. This study was undertaken to examine the serum uric acid levels and urine pH in rats after ambroxol parenteral treatment. Ambroxol influence on the uric acid level was measured in 5 rats (Rattus sp.) treated with 60 mg/kg (dissolved in injection water, sc, daily) during 2 weeks. Ambroxol influence on urine pH was examined on 45 rats divided into 3 groups. Rats from the 1st and 2nd group received 30 and 60 mg/kg/24h ambroxol, respectively. Urine was collected once daily and measured with strip kit. All values were presented as the means with standard deviations. The Student t test was used to compare the means, p < 0.05 was considered as significant. Dynamics of pH changes was measured in 4 rats treated with 60 mg/kg/24h of ambroxol. Controls received 1 mL of injection water sc. Serum uric acid level increased up to 8.7 +/- 1.0 mg/dL vs. 5.7 +/- 1.0 mg/dL in control (p < 0.002). In the 1st and 2nd group urine pH increased up to 7.5 +/- 0.5 and 7.6 +/- 0.5 vs. 6.7 +/- 0.4 (p < 0.05). Ambroxol withdrawal resulted in sequential urine pH decrease. 11 days after interruption of ambroxol therapy pH reached the starting value. Urine pH changes and possible disturbances in uric acid metabolic pathway may influence on the stone formation in rats after ambroxol parenteral treatment. The influence of ambroxol on urinary tract GAG layer and the balance between xanthine and CaOx in the urine should be checked.

Background: Surgical treatment of vitiligo is indicated when lesions are localized in poorly responding areas., Objectives: The objectives were: (1) to establish the melanocyte culture obtained from the epidermis of vitiligo patients for future treatment; (2) to estimate the influence of selected factors on the formation of suction blisters and the results of culture; and (3) to compare the results of treatment of vitiliginous macules localized in the dorsum of the hands and lower limbs by transplantation of cultured autologous melanocytes plus psoralen and ultraviolet A (PUVA) therapy (CMP), suction blister transplantation plus PUVA therapy (SBP), cryotherapy plus PUVA-therapy (CP), and only PUVA therapy (OP)., Methods: Forty patients were qualified for the study. The roofs of the suction blisters were used as a melanocyte source for culture establishment or were directly transplanted., Results: The CMP procedure was successfully performed on only 10 of 20 patients because of the difficulties in cell culture establishment. The SBP method was carried out on all 20 patients. A total lack of effectiveness was found in CP and OP methods., Conclusions: The effectiveness of culture depends on time of suction blister forming, phototype, and previous PUVA therapy. This study demonstrated the advantage of the SBP over the CMP method.

Liver transplantation is the only efficient method of treatment of liver failure. Short time between liver end-stage insufficiency and transplantation procedure is the main problem limiting the number of liver transplants. In this paper the methods of liver support in patients awaiting a liver transplant using tissue engineering techniques were introduced. The methods of liver support using bioartificial liver and isolated hepatocytes transplantation were described.

Objectives: Small intestinal submucosa forms a scaffold for tubular construction. The aim of this study was to build the artificial conduit using small intestinal submucosa (SIS) and 3T3 fibroblasts for urinary diversion in rats., Materials and Methods: 3T3 fibroblasts were multiplied to a total of 10(9). Two groups consisted of three Wistar rats each. The left ureters were separated from the bladder and anastomosed to the proximal end of the tubular scaffold. No splitting of the ureteral junction or drainage was done. The distal end of the scaffold was implanted into a previously performed channel in the abdominal wall. Cell-seeded grafts were used in the first group and acellular SIS scaffolds in the second group. Rats were sacrificed after 2 and 4 weeks. X-ray pyelography was performed. Hematoxylin and eosin staining was prepared from conduit cross sections., Results: All animals survived the observation. An inflammatory reaction was observed within the peritoneal cavity in both groups. It was difficult to dissect the adhesions in the cell-seeded group. The ureteral-conduit anastomoses were tight in five cases, except there was leakage and pseudocyst formation after 14 days in one cell-seeded graft. No ureterohydronephrosis was observed in two acellular conduits after 14 or 30 days, and in one case of a cell-seeded graft. A neovascularisation process was observed in the acellular conduit after a month. Multilayered epithelium covered the conduit lumen near the anastomosis at the distal end of acellular conduit, a small islet-forming epithelial layer was observed after a month., Conclusions: 3T3 fibroblasts cannot serve as a "feeder layer" for ureteral augmentation. It seems that there is no need to split the ureteral-conduit junction. An SIS scaffold was used for tubular construction for urinary diversion in an animal model.

Introduction: Ambroxol (ABX) is known to promote bronchial secretion and is used as an expectorant. This study was undertaken to document the connection between ambroxol parenteral treatment and bladder stones in rats., Material and Methods: Forty-five wild rats (Rattus sp.) were divided into three equal groups. Rats from the first and second groups received ABX s.c. during 2 weeks in total doses of 30 mg/kg per 24 h and 60 mg/kg per 24 h, respectively. Rats from the control group received 1 mL of injection solution s.c. One month after the treatment termination, animals were sacrificed and urinary tracts without urethra were dissected. Stones found in the bladders were measured, weighed and chemically analysed. Voiding cystography was performed to exclude pathology of the lower urinary tract. Photo documentation was produced., Results: From the first and second groups, 33% and 47% of rats, respectively, had solitary stones in the bladder. In one case from the second group, there was a huge stone in the bladder and urethra. There were no stones in rats from the control group. The mean length of stones was 1.38 +/- 0.23 mm and 1.41 +/- 0.60 mm in the first and second groups, respectively. Mean stone weight was 1.2 +/- 0.2 x 10(-3) g and 1.44 +/- 0.54 x 10(-3) g. Stones were composed of 67% of xanthine and 33% of calcium oxalate., Conclusions: Ambroxol parenteral treatment caused xanthine and oxalate stone formation. Attention should be paid to the possibility of urinary stone formation after long-term ABX treatment.

There is an increasing interest in using of silver coated (silver nitrate, silver alloy, silver oxide) catheters for hospital patients. Clinical trials with silver-coated urinary catheters have shown conflicting results. The most often performed catheterization is for a short period of time. The above observations have encouraged the authors to investigate the influence of silver nitrate (AgNO3) on 3T3 fibroblasts viability in vitro during a short time experiment (3 and 12 h). 3T3 fibroblast culture was established. The influence of AgNO3 on the viability of murine 3T3 fibroblasts with the use of trypan blue staining was evaluated. The regression curves and lethal concentrations for 90, 50 and 10% viability were calculated. The lethal concentrations of AgNO3 after 3 h exposition were as follows LC10=0.98, LC50=6.44 and LC90=21.38. The lethal concentrations of AgNO3 after 12 h exposition were as follows LC10=1.05, LC50=6.91 and LC90=22.96. The LC values were similar for 3 and 12 h exposure as well. In conclusion, the silver nitrate has the similar toxic effect on 3T3 fibroblasts during the short and long exposition. Attention should be paid when catheter has a close contact to injured urothelium even for a short period of time.

The adult stem cells (ASC) are supposed to regenerate epithelium. We hypothesized prostate epithelial CD133-positive ASC to be responsible for establishing the primary cell culture. The prostate epithelial stem cells were isolated using anti-CD133 microbeads in order to form different cell populations. The morphology of cultures developed from CD133(+) and CD133(-) prostate epithelial cells were compared with prostate epithelium cell culture obtained after simple isolation procedure. Four 8-week-old Wistar rats were used in the experiment and six cultures were obtained. Double CD133(+) and CD133(-) cultures from two rats were established after enzymatic digestion and positive selection by SuperMACS device, and two non-selected CD133(+)/CD133(-) cultures were developed by simple prostate epithelial cell isolation from two other rats. The epithelial nature was confirmed by anti-cytokeratine antibodies. It was observed that growth of the CD133(+)/CD133(-) and CD133(+)culture resembled epithelial-like prostate cell culture. It was not possible to establish epithelial-like culture from CD133(-) cell population. The primary epithelial cell culture collapsed in a few days after the CD133-positive ASC were removed. We concluded that the epithelial progenitor cells are responsible for establishing primary prostate epithelial cultures in vitro.

Purpose of Investigation: The aim of the study was to determine the effect of different types of management on the activity of catalase (CAT) and glutathione peroxidase (GPx) in women with cervical carcinoma., Methods: The patients were divided into three groups according to the mode of treatment. Patients from the first group were treated brachytherapy prior to surgery. The second group received teletherapy before brachytherapy and additionally chemotherapy. The third group was treated with teletherapy after brachytherapy sessions., Results: CAT activity was higher while GPx activity was lower before and during therapy in all groups as compared to controls. Six months after the end of therapy, the activity of studied enzymes reached the values characteristic of healthy women. No significant differences in enzyme activity among the three groups were revealed., Conclusion: Normalization of CAT and GPx activity may prove the efficacy of applied therapy in cervical cancer patients, however enzyme activity recovery was not dependent on treatment mode.

The aim of this study was to evaluate the concentration of malondialdehyde-MDA (one of the lipid peroxidation products)-in blood plasma and erythrocytes and the activity of superoxide dismutase (SOD) and catalase (CAT) in red blood cells of patients with a primary brain tumour. The study was performed on 24 patients with a brain tumour (9 with glioblastoma and 15 with asrocytoma) treated in the Department and Clinic of Neurosurgery and Neurotraumatology at Ludwik Rydygier Medical University in Bydgoszcz. The control group consisted of 20 healthy volunteers. A statistically significant higher MDA concentration in erythrocytes and blood plasma and a higher activity of SOD or CAT in erythrocytes was shown in patients with a brain tumour as compared to the control group. Neither the histological type of tumour nor surgery has an effect on the tested biochemical parameters.

Stevens-Johnson syndrome (SJS) is a mucocutaneous disorder induced by an immune-complex-mediated hypersensitivity reaction. Nearly half of cases are caused by a reaction to drugs or appear during viral infections and malignancies. A very few cases are caused by a bacterial infection (Streptococcus) or Mycoplasma pneumoniae. Graft versus host disease is another well-established cause, independent of drugs. No specific etiology has been identified in up to half of cases. We report a 54-year-old man with SJS induced by carbamazepine. Reported patient had prodromal symptoms like fever, headache and polyarthralgia, which preceded mucocutaneous lesions by 3 days. Physical examination on admission, revealed asthenic male with a temperature of 37.2 degrees C and generalized dermatitis with positive Nikolsky sign, large erosions of the palms and soles, onychomadesis, numerous oral and vermilion border of the lips erosions. The patient was administered systemic steroidotherapy and carbamazepine dose was gradually decreased and finally replaced with valproic acid and valproate sodium. During the hospitalization, temperature normalized and the skin lesions disappeared after 3 weeks of treatment.

Cancer stem cells are a small subset of cancer cells constituting a reservoir of self-sustaining cells with the exclusive ability to self-renew and maintain the tumor. These cells are identified by specific stem cell markers: antigens, molecules and signaling pathways. Transcription factors and molecules associated with oncogenesis, such as NF-kappaB, Bmi-1, Notch, WNT beta-catenin, Sonic hedgehog and their biochemical pathways, active only in a small minority of cancer cells might play key roles in determining the biology and the overall long-term behavior of a tumor. The molecules and pathways specific for cancer stem cells, which contribute to their drug resistance, are potential targets for new therapeutic strategies.

Objectives: Fibroblasts sometimes occur after enzymatic isolation of epidermis. They proliferate quickly, overgrowing the culture. A pure epithelial culture is essential for therapy using a keratinocyte graft. The aim of this study was to determine the possibility of fibroblast elimination from culture to prevent fibroblast overgrowth and obtain a pure monolayer of keratinocytes., Material and Methods: We analyzed three epidermal-derived cultures. Cells were cultured in medium contained Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 at a 3:1 ratio with 5% autologous serum and additives. The epithelial culture was confirmed using pancytokeratin MMF. If fibroblast like cells were present, they were removed using 0.01% edetate disodium dihydrate (Na2EDDA). This procedure was repeated until we obtained pure primary keratinocyte cultures., Results: Fibroblast detachment was observed after Na2EDDA treatment. The procedure was performed twice and pure primary cultures of keratinocyte were achieved in two cases. These two cultures maintained their epithelial-like morphology and cytokeratin expression. One culture was treated four times with Na2EDDA with no effect; the morphology of the cultures became fibroblast-like with no observed cytokeratin expression., Conclusions: Unwanted dermal fibroblasts can be separated from primary keratinocyte cultures during the first few days after the isolation. Cocultures of unwanted dermal fibroblasts and epidermal keratinocytes can be reverted to pure keratinocyte monolayers suitable as grafts for transplantation.

Staphylococcin T (StT) is a bacteriocin, which can serve as an antibiotic. The influence of StT on the mammalian cells viability should be examined before its possible applications. The aim of this work was to study the influence of StT on mouse fibroblasts viability in vitro. StT was delivered as a deposit in agar. 3T3 fibroblasts were used in viability tests. Cells were incubated with StT for 24 and 48 h. In the first controls, cells grew without the agar block, in the second controls, the agar block contained no StT. Trypan blue exclusion test was performed. Growth inhibition of sensitive strain SAU-209P was observed after 24 h incubation with cell culture medium containing StT. Antimicrobial activity of StT in culture medium was confirmed even after 2 months. The viability of 3T3 cells in the second control group was 50% lower than in the first and 25% higher than in StT treated group during the first day. It was noticed mechanical damage of growing cells caused by agar blocks. The number of cells cultured with StT was similar to that of second control group after 2 days. There were no changes in morphology of cells in all groups. It follows that StT had unimportant influence on the fibroblasts (3T3) viability in culture. StT may be considered for future animal trails.

An aim of the study was to determine the protein expression of the FAS-related apoptosis signaling pathway (FADD-FAS Associating Protein with Death Domain, PRO-CASPASE-8 and CASPASE-8), which are responsible for signal transduction to trigger programmed cell death (apoptosis) in cancer and Prostatic Intraepitelial Neoplasia (PIN). 20 specimens from prostate cancer patients treated with radical prostatectomy were inwestigated. 8 cancers were diagnosed as G-2 and 12 as G-3. 14 samples were described as poorly differentiated, high Gleason score (> or = 7). Control group consisted of prostate specimens from autopsy of 3 young men. Specimens were fixed in 10% buffered formaldehyde and immersed in paraffin. Haematoxylin and eosin staining was done. Monoclonal antibodies to FADD & CASPASE-8 (Novocastra, UK) were used to immunohistochemical study, according to streptavidine-biotin method. Semiquantitive method described protein expression. Expression index (EI) was calculated as a percent of positive FADD or CASPASE-8 cells to total cells in the specimen. Statistical analysis was performed with the Student t-test (p < 0.05). Normal prostate tissue was negative in both, FADD and CASPASE-8 immunohistochemistry staining. PIN & prostate cancer lesions were found to strongly express of FADD & CASPASE-8 proteins. Expression of FADD in cancer lesions was 66,5+/-27,8% and 59,8+/-19,0% vs. 56,8114,8% HGPIN and LGPIN, respectively. Expression of CASPASE-8 in cancer lesions was 64,1 + 23,4% and 61,5+/-15,0% vs. 48,0+/-17,6% HGPIN and LGPIN, respectively. PIN & prostate cancer lesions are characterized by similar high expression of proteins responsible for signal transduction to induce apoptosis. The mediators of apoptotic signal can be very important in prostate cancer prophylaxis and management.

Treatment of diabetes mellitus poses a huge medical problem. Both the transplantation of pancreas and insulin treatment is connected with numerous complications. The method of treatment which would entirely relieve patients from the necessity of insulin applications and complications of diabetes mellitus has yet not been found. The paper addresses to the issues related to procurement, preservation and culture of pancreatic islets. In this review, the current state of knowledge as well as perspectives of treatment of diabetes through the transplantation of insulin secreting cells was presented.

Unlabelled: There is no ideal culture establishing method for the conjunctival sac reconstruction. Two methods of primary conjunctival culture establishing were compared., Materials and Methods: The internal surface of rat's eyelid was used. In the 1st method cells migrate from specimen and attached to the surface of culture plate. In the 2nd method tissue was digested and placed in the flask. Epithelial character of cultured cells was confirmed., Results: Both methods allow to set up the culture of epithelial cells. Efficiency of enzymatic isolation was higher then growing from previously attached specimen. At the end of the experiment the cultures were similar., Conclusions: Both methods allow to establish the primary culture of conjunctival epithelial cells. Eyelid conjunctiva can be a cells source for the conjunctival sack reconstruction purpose.

The etiology of the interstitial cystitis (IC) is believed to be a dysfunction of the bladder glycosaminoglycan (GAG) protective layer of the urothelial cells. Management of interstitial cystitis (IC) is pure empirical. The aim of this study was to establish primary rabbit urothelial cells (PRUCC) affinity to complex sugar scaffold in vitro. The primary rabbit urothelial cells culture (PRUCC) was set up. The cells were cultured in Jouan IG 150 incubator in temperature of 370C and humidified atmosphere containing 5% of CO2. The multi-well culture plates were covered with potato starch. PRUCC was seeded on the starch scaffold. On the 7th day the starch was removed. Culture analysis and photo documentation was done under inverted microscope. There was no disruption of the monolayer under the starch and the cells morphology was comparable to the rabbit urothelial cells in culture. No inhibitory effect on the PRUCC was observed. The colonies of the urothelial cells were formed over the starch grains. The athor postulate that urothelial cells have affinity to the complex sugar compounds. There is strong evidence that sugar complex compounds could be used in the treatment of the IC.