Rare neoplasm of genito-urinary tract. Genito-urinary sarcomas: chemotherapy.

PURPOSE OF REVIEW: The purpose of this article was to describe the systemic therapy of genito-urinary sarcomas. RECENT FINDINGS: High rate of distant metastasis and high mortality rate has brought interest into the development of new therapeutic approaches. Various modules of chemotherapy were sampled in sarcoma treatment, although clinical response is still unsatisfactory. Chemotherapy in sarcomas can be used as neoadjuvant or adjuvant to the surgery. There is no consensus on the current role of adjuvant chemotherapy. Study results are conflicting; therefore, conclusions drawn from the studies are uncertain. In general, the adjuvant chemotherapy is not standard treatment in adult-type sarcomas. In addition, chemotherapy for advanced and metastatic sarcoma disease, as well as second-line chemotherapy, was discussed. SUMMARY: The best treatment for sarcomas in case of organ-confined disease and in selected cases of locally advanced disease seems to be surgery followed by chemotherapy. In case of metastasis stage of sarcoma, preoperative chemotherapy with surgery of residual masses should be considered as first-line treatment followed by postoperative chemotherapy. The treatment of patient with highly advanced disease and/or unresectable metastases should be individualized (chemotherapy, radiotherapy and best supportive care).

The role of early diagnosis of emphysematous cystitis: A case report and literature review.

Emphysematous cystitis (EC) is a rare entity caused by bacteria, which produce gas filled cysts in the bladder wall. We present a case of EC in a 72-year-old woman admitted to Vascular Surgery Department because of diabetic foot syndrome. During the hospital stay, the patient's general condition deteriorated. CT established EC diagnosis. Surgical treatment was inevitable. Salvage cystectomy was performed. Despite macroscopic removal of necrotic tissues, the condition of the patient didn't improve, 75 days past diagnosis of EC she died due to the multi-organ failure. Prompt diagnosis provided by imaging plays a key role in the treatment of EC.

Mumio (Shilajit) as a potential chemotherapeutic for the urinary bladder cancer treatment.

Mumio (Shilajit) is a traditional medicinal drug known and used for hundreds of years. Bladder cancer is one of the most common cancer types and better treatments are needed. This study analysed the in vitro effect of Mumio on urinary bladder cancer cells (T24 and 5637) in comparison to normal uroepithelial cells (SV-HUC1). Cytotoxicity of Mumio was analysed in these cell lines via MTT and real-time cell growth assays as well via the assessment of the cytoskeleton, apoptosis, and cell cycle. Mumio affected the viability of both cell types in a time and concentration dependent manner. We observed a selectivity of Mumio against cancer cells. Cell cycle and apoptosis analysis showed that Mumio inhibited G0/G1 or S phase cell cycle, which in turn induced apoptosis. Our results showed that Mumio was significantly more cytotoxic to urinary bladder cancer cells than to normal cells. These results are promising and indicate Mumio as a great candidate for urinary bladder cancer treatment and further investigations should be performed.

The development of marine biomaterial derived from decellularized squid mantle for potential application as tissue engineered urinary conduit.

Tissue engineering is focusing research effort on search for new biomaterials that might be applied to create artificial urinary conduit. Nevertheless, the demanding biomechanical characteristics necessary for proper conduit function is difficult to be replicated. In this study, we are introducing novel marine biomaterial obtained by decellularization of squid mantle derived from Loligo vulgaris. Squid mantles underwent decellularization according to developed dynamic flow two-staged procedure. Efficacy of the method was confirmed by computational dynamic flow analysis. Subsequently Decellularized Squid Mantle (DSM) underwent extensive histological analysis and mechanical evaluation. Based on gained biomechanical data the computational modelling using finite element method was utilized to simulate behavior of DSM used as a urinary conduit. Taking into account potential application in reconstructive urology, the DSM was then evaluated as a scaffold for urothelial and smooth muscle cells derived from porcine urinary bladder. Conducted analysis showed that DSM created favorable environment for cells growth. In addition, due to polarized structure and natural external polysaccharide layer, it protected seeded cells from urine.

Development of a conductive biocomposite combining graphene and amniotic membrane for replacement of the neuronal network of tissue-engineered urinary bladder.

Tissue engineering allows to combine biomaterials and seeded cells to experimentally replace urinary bladder wall. The normal bladder wall however, includes branched neuronal network propagating signals which regulate urine storage and voiding. In this study we introduced a novel biocomposite built from amniotic membrane (Am) and graphene which created interface between cells and external stimuli replacing neuronal network. Graphene layers were transferred without modifying Am surface. Applied method allowed to preserve the unique bioactive characteristic of Am. Tissue engineered constructs composed from biocomposite seeded with smooth muscle cells (SMC) derived from porcine detrusor and porcine urothelial cells (UC) were used to evaluate properties of developed biomaterial. The presence of graphene layer significantly increased electrical conductivity of biocomposite. UCs and SMCs showed an organized growth pattern on graphene covered surfaces. Electrical filed stimulation (EFS) applied in vitro led additionally to increased SMCs growth and linear arrangement. 3D printed chamber equipped with 3D printed graphene based electrodes was fabricated to deliver EFS and record pressure changes caused by contracting SMCs seeded biocomposite. Observed contractile response indicated on effective SMCs stimulation mediated by graphene layer which constituted efficient cell to biomaterial interface.

Alginate is not a good material for growth of rapidly proliferating cells.

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.

Bone marrow progenitors from animals with chronic renal failure lack capacity of in vitro proliferation.

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.

The effect of combined therapy on activity of cathepsin D and alpha-1-antitrypsin in the blood serum of women with cervical cancer.

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.

The artificial conduit for urinary diversion in rats: a preliminary study.

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.

Influence of the management of cervical carcinoma on the activity of catalase and glutathione peroxidase in erythrocytes.

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.

Scaffold seeded with cells is essential in urothelium regeneration and tissue remodeling in vivo after bladder augmentation using in vitro engineered graft.

OBJECTIVE: Tissue-engineering methods using synthetic biodegradable scaffolds seeded with cells have potential to induce regeneration to a functional bladder wall. The aim of the study was to induce in vivo urothelial growth on implanted scaffolds previously seeded with stromal cells as compared with matrices implanted without cells for rat cystoplasty augmentation. MATERIALS AND METHODS: 3T3 mouse fibroblasts were multiplied up to total of 10(8) cells. Cells were grown on Dulbecco's modified essential medium supplemented with 10% of fetal bovine serum and antibiotics in CO(2) chambers. Cells were seeded on biodegradable polyglycolic acid (PGA) scaffolds in eight rats: four bladders were augmented with cell-seeded grafts and the other four with acellular scaffolds. Rats were sacrificed after 4 months in preparation for hematoxylin and eosin staining. RESULTS: One death in the acellular cystoplasty group was observed after 3 weeks. No epithelial layer was observed in the central part of the acellular graft. The cell-seeded grafts showed good visible multilayered epithelium with at least five layers of epithelial cells in the central part. The epithelium resembled rat native urothelium. The cell-seeded grafts showed a high degree of implanted 3T3 cells infiltration with good degradation of PGA fibers. CONCLUSIONS: Our data indicated that urothelial proliferation on PGA grafts was intensified using a "feeder layer" of fibroblasts.

Does the presence of unwanted dermal fibroblasts limit the usefulness of autologous epidermal keratinocyte grafts?

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.

Kidney preserving solutions containing lidocaine may increase urological complication rate after renal transplantation: an in vitro study.

OBJECTIVE: The frequency of urological complications after renal transplantation is up to 12%. Some authors consider that lidocaine addition to preservation solutions produces a favorable influence on allograft function. However, lidocaine may influence urinary tract epithelial cells. The aim of this work was to establish the influence of lidocaine on cultured primary rabbit urothelial cells (PRUC) as a tool to understand mechanisms of urological complications after kidney transplantation. DESIGN AND METHODS: A PRUC culture was established from an 8-month-old male rabbit bladder. The cells were cultured alone and then with in various concentrations of lidocaine for 24 hours or 1 hour. After an additional 24 hours, cell viability was assessed by the trypan blue exclusion test. Student's t test was used for statistical analysis, with significance set at P < .05. RESULTS: The cytotoxic effects of lidocaine on PRUC were concentration dependent. One-hour exposure of PRUC culture to 0.5 or 1.0% lidocaine decreased cell viability. Both lidocaine concentrations decreased cell viability in PRUC culture after a 24-hour incubation; even 0.25% lidocaine caused changes in the PRUC culture morphology after a 1-hour incubation. Cells became rounded and detached from the growth surface. No cells were observed in the monolayer after 1-hour incubation with 1% of lidocaine. CONCLUSIONS: The toxic effects of lidocaine on PRUC may forecast problems with supplementation of kidney preservation solutions, leading to impaired epithelial layer healing with an increased risk of urological complications.

Activity of superoxide dismutase (SOD) and concentration of thiobarbituric acid reactive substances (TBARS) in liver and muscles of some fish.

In this study we examined superoxide dismutase (SOD) activity and thiobarbituric acid reactive substances (TBARS) concentration in liver and muscles of four fish species: the carp, the brown trout, the white cod and the flounder. Higher SOD activity and higher TBARS concentration was revealed in the tissues of marine fish in comparison to freshwater fish. The highest SOD activity was observed in the cod while the highest TBARS concentration was in the flounder. The observed differences are probably an effect of the different living mode of the compared fish.

Urine--a waste or the future of regenerative medicine?

In recent years, urine has emerged as a source of urine cells. Two different types of cells can be isolated from urine: urine derived stem cells (USCs) and renal tubular cells called urine cells (UCs). USCs have great differentiation properties and can be potentially used in genitourinary tract regeneration. Within this paper, we attempt to demonstrate that such as easily accessible source of cells, collected during completely non-invasive procedures, can be better utilized. Cells derived from urine can be isolated, stored, and used for the creation of urine stem cell banks. In the future, urine holds great potential to become a main source of cells for tissue engineering and regenerative medicine.

New application of carbon nanotubes in haemostatic dressing filled with anticancer substance.

The drug-carrier system used as innovative haemostatic dressing with oncostatic action is studied. It is obtained from CDDP (cisplatin) doped SWCNT (single walled carbon nanotubes), modified and purified by H2O2 in hydrothermal treatment process. In the in vivo nephron sparing surgery (NSS) study we used 35 BALB/c nude mice with induced renal cancer using adenocarcinoma 786-o cells. Animals were divided into four groups: CDDP(M-), CDDP(M+), CONTROL(M-) and CONTROL(M+). In CDDP(M-) and CDDP(M+) groups we used, intraoperatively, carbon nanotubes filled with cisplatin (CDDP). In CONTROL(M-) and CONTROL(M+) groups carbon nanotubes were used alone. During NSS free margin (M-) or positive margin (M+) was performed. In the CDDP(M-) group, we do not observe local tumor recurrences. In Group CDDP(M+) only one animal was diagnosed with tumor recurrence. In control groups the recurrent tumor formation was observed. In our study, it is shown that CDDP filled SWCNT inhibit cancer recurrence in animal model NSS study, and can be successfully applied as haemostatic dressings for local chemoprevention.

Lipid peroxidation and antioxidant capacity in selected tissues of healthy black C57BL/6J mice and B16 melanoma-bearing mice.

During the process of melanogenesis free radicals are generated. The aim of this study was to investigate the effects of melanogenesis in B16 melanoma on lipid peroxidation and antioxidant capacity in selected tissues of black C57BL/6J mice. The study was conducted on 24 mice: 12 healthy controls and 12 with a transplanted B16 melanoma. Two weeks after the melanoma transplant, when the average weight of the tumours was approximately 2.0 g, blood samples were taken from the orbital venous plexus. The mice were killed by dislocation of the spinal cord, and the brain, liver and lungs were removed for analysis. The level of thiobarbituric acid-reactive substances (TBARS) and of 1,1-diphenyl-2-picrylhydrazyl (DPPH) reactive substances were determined in full liver, lung and brain homogenates and in serum. The activity of superoxide dismutase (SOD) was determined only in homogenized tissue. The concentration of TBARS and the SOD activity were statistically significantly higher in all the studied tissues from mice with B16 melanoma than in tissues from healthy mice. The antioxidant capacity, however, was lower in the tissues of melanoma-bearing mice. The results obtained demonstrate an increase in oxidative stress in the tissues of mice bearing a transplanted B16 melanoma.

Activity of antioxidant enzymes and concentrations of thiobarbituric acid reactive substances (TBARS) in melanotic and amelanotic Bomirski melanoma tissues in the golden hamster (Mesocricetus auratus, Waterhouse).

The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSHPx), as well as the concentration of thiobarbituric acid reactive substances (TBARS) in tissues of transplantable melanoma in the golden hamster were measured and compared. Ten inbred male hamsters were used for the experiment. They were divided into two groups and were given Bomirski melanoma cells subcutaneously. The first group was given melanotic (Ma) melanoma cells. The second group was given amelanotic (Ab) melanoma cells. Thirty days after the transplantation the hamsters were dissected and the tumor tissues were taken and homogenized. A statistically significantly higher activity of the measured antioxidant enzymes was found in homogenates of Ma tumor than in homogenates of the Ab tumor. Activity of SOD is 8% higher in melanotic melanoma, 24% higher in CAT, and 45% higher in GSHPx. Statistically significant differences between TBARS concentrations were not confirmed. The higher activity of antioxidant enzymes in the melanotic tumor is a result of increased generation of oxygen-derived free radicals. It is presumed that it is strictly connected with intensified production of quinone and semiquinone radicals in the process of melanogenesis.

Activity of some lysosomal enzymes in serum and in tumors of patients with squamous cell lung carcinoma.

The aim of the study was an assessment of some lysosomal enzymes activity in serum and in tumors of patients with lung cancer histopathologically confirmed as squamous cell lung carcinoma. The first group constisted of 10 patients with stage II of the disease and the second group consisted of 11 patients with stage III of the disease. Lysosomal enzymes activities were assayed in serum before surgery and on the 10th day after surgery in serum and in tumors. Arylsuphatase, cathepsin D and acid phosphatase activities were higher in the patients serum than in that of the control group. The decrease of arylsulphatase and cathepsin D activities after surgery was statistically significant in both groups of patients, but the cathepsin D activity was still 3 times higher in patients than in those from the control group. The decrease of acid phosphatase activity after surgery was about 50% in both groups of patients and this decrease was statistically significant. The arylsulphatase and acid phosphatase activity in tumors was nearly 3 times higher in stage III patients than it was in stage II patients, but the cathepsin D activity was nearly the same in both patient groups. Higher lysosomal enzyme activity may be a useful factor in diagnosing and monitoring of lung cancer. However, further investigations are needed.

The in vitro study of influence of four novel platinum compounds on rodent melanoma cells.

The purpose of this study was to assess cytotoxic effect of four new platinum compounds on B16 and CIS91 melanoma cells in vitro. The following complexes were tested: (2) Tetrachlorobis (5,7-dimethyl-1,2,4-triazol [1,5alpha] pirymidine) platinum (IV), (3) trans-dichloro (dimethylsulfoxide) (5,7-dimetyl-1,2,4-triazol-[1,5alpha] pirymidine) platinum(II), (4) cis-dichloro(dimethylsulfoxide)(1-beta-D-ribofuranosyl-1,2,4-triazol-3-carboxamide)platinum(II), (5) chloro(dimethylsulfoxide)(1-beta-D-ribofuranosyl-1,2,4-triazol-3-carboxamide)platinum(II). We can conclude, that Pt-dmtp (2) represented the best cytotoxic properties in the group of four tested platinum compounds, Pt-rib-1 (4) has also a good cytotoxic properties although its IC50 value is quite high. We suppose that cytotoxic and soluble properties ot Pt-dmtp and Pt-rib-1 could be modified and improved.