Marek Lipiński

Alternating expression levels of WWOX tumor suppressor and cancer-related genes in patients with bladder cancer

The aim of the present study was to determine the roles of the WWOX tumor suppressor and cancer-related genes in bladder tumor carcinogenesis. Reverse transcription-quantitative polymerase chain reaction was used to analyze the status of WWOX promoter methylation (using MethylScreen™ technology) and loss of heterozygosity (LOH) in papillary urothelial cancer tissues. The associations between the expression levels of the following tumorigenesis-related genes were also assessed: The WWOX tumor suppressor gene, the MKI67 proliferation gene, the BAX, BCL2 and BIRC5 apoptotic genes, the EGFR signal transduction gene, the VEGF vascular endothelial growth factor gene, and the CCND1 and CCNE1 cell cycle genes. The results reveal a high frequency of LOH in intron 1 in the WWOX gene, as well as an association between reduced WWOX expression levels and increased promoter methylation. In addition, the present study demonstrates that in bladder tumors, apoptosis is inhibited by increased expression levels of the BCL2 gene. A correlation between the proliferation indices of the MKI67 and the BIRC5 genes was also revealed. Furthermore, the expression levels of VEGF were identified to be positively associated with those of the EGFR gene.

Complication rates after prostate biopsy according to the number of sampled cores.

Introduction A prostate biopsy can result in such complications as: hematuria, rectal bleeding, pain in hypogastrium, perineum or urethra, fever, nausea, vomiting, retention of urine or other adverse events. The aim of this research was to estimate complication rates after a prostate biopsy based on the number of cores. Material and methods The complication rate was evaluated on the basis of questionnaires filled out by patients. Questions were related to the occurrence of mentioned complications on the first and second day after prostate biopsy. Patients were divided into two groups: 1st group (41 patients) 5-8 cores and 2nd group (73 patients) 12 or more cores. Results There was no significant statistical difference in the occurrence of complications mentioned in the questionnaires in both groups. The biggest difference was recorded for hematuria – 1st day: 39% in the 1st and 53% in the 2nd group (p = 0.1398); 2nd day: 15% in the 1st and 30% in the 2nd group (p = 0.0650). Rectal bleeding on the 1st day also seems to vary: 12% in the 1st and 26% in the 2nd group (p = 0.0835). Other complications occurred in 3-8% of patients. 32% of patients in the 1st and 29% in the 2nd group (p = 0.7419) had no complications at all. Conclusions The most common complications after a prostate biopsy are hematuria and rectal bleeding. Other complication rates are low. In general, complication rates after a prostate biopsy procedure are not related to the number of sampled cores.

Rate of positive urine culture and double–J catheters colonization on the basis of microorganism DNA analysis

Introduction The aim of the trial was to estimate the relationship between colonization of the Double–J catheter, and the microorganisms cultured from urine. Material and methods 60 patients, who had Double–J catheters inserted, participated in the study. All the subjects had their midstream urine samples taken prior to the stent insertion and removal. A negative urine culture before catheterization was mandatory to participate in the study. The patients were assigned into three subgroups, according to stenting duration: 1) 20 to 30 days (18 cases); 2) 30 to 90 days (30 cases); 3) longer than 90 days (12 cases). Bacterial and fungal DNA was identified using electrophoresis in polyacrylamide gel with a denaturing gradient (PCR–DGGE). The relationship between the genetic analysis of the catheter and the urine culture was estimated. Results Urine cultures were positive in only 8 patients, while Double–J catheter analyses were positive in all cases. In 2 cases one type of microorganism was isolated from the stent surface while the remaining 58 catheters were colonized by more than one pathogen. In three cases fungi were isolated. There were only three types of pathogens cultured from urine specimens. Urine and stent cultures were consistent in 5 cases. In 3 cases urine culture and stent analysis were not consistent. Conclusions Double–J catheter retention in the urinary tract is associated with an extremely high risk of bacterial colonization, while the risk of urine infection is about 8–fold lower. There is a great inconsistency between urine infection and catheter colonization, indicating a low predictive value of urine culture for estimating stent colonization.

Semen quality in men from subfertile couples from the region of Łódź (Poland) according to the new reference values recommended by WHO 2010.

The semen analysis is the main diagnostic tool for evaluating the male fertility potential. The standard semen analysis includes evaluation of the sperm concentration, motility, and their morphology. The most important question is whether the results from semen analysis may be accurate markers for male fertility. Therefore, we retrospectively studied sperm quality among men attending the infertility clinic due to reproductive problems consistent with the WHO manual from 1999, which were reassessed according to the manual from 2010. Semen results from 571 males from couples undergoing fertility investigation were analyzed. All subjects included in the study had no abnormalities during examination. In 64 samples (11.2%), a leukocyte count above 1 x 106/ml was found and their semen volume (median 3.2 ml) was significantly lower in comparison with the group without leukocytes (3.6 ml; p <0.001). Normal semen parameters were found in 290 subjects (50.8%) according to the 1999 manual and in 362 men (63.4%) according to the 2010 manual. The normozoospermia group, according to the 2010 manual, had a significantly lower percentage of sperm with progressive motility, motile sperm concentration, and total number of motile sperm in comparison with the normozoospermia group according to the manual from 1999. It seems that routine semen analysis is not sufficient to estimate male fertility potential and some men with normal semen parameters may be subfertile. Further investigations are needed.

Comparison of the effectiveness of crushing concrements in the urinary tract with the use of holmium laser and sonotrode.

Introduction Holmium lasers and ultrasound probes are widely used in urinary stone lithotripsy. The authors present a comparison of both methods in urinary stones lithotripsy. Materials and methods We analyzed 164-patients who underwent endoscopic lithotripsy. Ninety-eight of them (group I) were treated with the holmium laser Omni-Pulse Max 80 and were compared to the other 66-patients (group II) who were operated on with an ultrasound probe (sonotrode). Prior to the procedures, all patients were exposed to urological ultrasound and radiological imaging in order to localize the stone. The state where the stones were invisible in the radiological and renal ultrasound imaging that was performed after the operation was considered to be effective. Patients were operated under intravenous general or subepidural anesthesia. Results The effectiveness of laser lithotripsy in the bladder amounted to 100% (25 of 25 patients). In the case of stones localized in the ureter it reached 89%. The total effectiveness of the procedure reached 92% (90 out of 98 procedures). In group II, the total effectiveness of the procedure reached 79% (52 out of 66 procedures). In group I, two cases the inflammatory changes of the ureter made the endoscopy impossible. These two patients were operated with open surgery. In one case, the laser lithotripsy was complicated with a perforation of the ureter. Conclusion The results prove that laser lithotripsy is a method of high effectiveness with a low risk of complication. It might surpass sonotrode and become its alternative. Both methods have both advantages and disadvantages.

Photodynamic diagnosis – current tool in diagnosis of carcinoma in situ of the urinary bladder

Carcinoma in situ (CIS) of the urinary bladder is extremely hard to diagnose. The symptoms are highly unspecific and the small, flat CIS lesions can easily be missed, thus remaining unseen in standard white light cystoscopy. Photodynamic diagnosis (PDD) is recommended by the European Association of Urology (EAU) as a diagnostic procedure in cases of suspected CIS [1]. A 58-year-old man visited the outpatient clinic complaining of symptoms of dysuria and occasional pain during micturition. The patient had been a smoker for 20 years, with no risk of exposure to chemical substances. The medical history did not indicate any haematuria or fever. Digital rectal examination found the prostate to be of proper density, with little enlargement and no other pathological findings. The preliminary diagnosis was set as lower urinary tract syndrome secondary to prostate enlargement. The patient was sent for creatinine assay, prostate specific antigen (PSA) assay, a urine test, and ultrasound examination of the kidneys, bladder, and prostate. He received a prescription for diclofenac, tamsulosin, and ciprofloxacin. The next appointment in the outpatient clinic was set for two months later as a follow-up. During that visit, the patient reported two episodes of painless bleeding in his urine. The serum levels of creatinine and PSA were 0.9 and 1.2 ng/ml, respectively, and no abnormalities were observed in the results of the urine test. No pathological changes were observed by ultrasonography. After the treatment, the symptoms of dysuria were diminished. In response to the presence of bleeding in the urine, the patient was sent for urethrocystoscopy, performed with short intravenous anaesthesia, and urinary cytology, which revealed the presence of pathological cells. On the basis of this result and that of the earlier ultrasonography, which indicated no bladder tumour symptoms, the patient was qualified for PDD. Photodynamic diagnosis was performed after intravesical instillation of a photosensitiser (Hexvix®) 60 minutes before the procedure. A Wolf PDD system was used to observe the photodynamic effect. Although the white light cystoscopy revealed only one pathological lesion, 4 mm in diameter, on the front wall of the urinary bladder, the urinary bladder mucosa seemed otherwise unaffected by any pathological changes. After switching to PDD mode, three independent points were indicated by red flashes: one on the back wall (8 mm in diameter), one on the right wall (5 mm in diameter), and one on the left wall of the urinary bladder (5 mm in diameter) (photo). Tissue samples were taken from each of the suspicious lesions. Both the papillary tumour and the three lesions visible only in PDD were treated by transurethral resection of the bladder tumour (TURBT). After the operation, instillation of mitomycin C was performed. The histopathological report revealed the papillary tumour from the front wall to be a high-grade non-muscle-invasive bladder cancer (G2) without lamina propria infiltration, the lesions on the back and left walls were found to be carcinoma in situ, and the one on the right wall was a high-grade dysplasia. The patient was immediately qualified to immunotherapy with BCG. In common practice, the diagnosis of urinary bladder CIS with non-specific symptoms often presents a problem [2]. Although urinary cytology has high sensitivity for CIS in cases such as specific and non-specific inflammatory processes in the urinary bladder, it can give false positive results. In addition, the result is highly dependent on the experience of the pathologist [3, 4]. Although CIS can appear to be a flat lesion in white light cystoscopy, with oedema of the urothelium and with abnormal blood vessel patterns, there is, generally speaking, no single standard picture of CIS. Hence, there is a strong need for diagnostic tools that can be used to visualise CIS when it is presented in the urinary bladder. Photodynamic diagnosis is a tool that can be used in the diagnosis of CIS as part of the TURBT (Transurethral Reaction of Bladder Tumour) procedure and re-TURBT, particularly in combination with Hexvix [5]. In a multicentre study on 286 patients, PDD examination identified CIS lesions in 80 patients (96%), and white light cystoscopy allowed for identification of CIS in 64 patients (80%) [6]. Another multicentre study conducted in the USA also compared standard white light cystoscopy and PDD in a group of 311 patients. Statistical analysis was performed on 196 patients. CIS was diagnosed in 56 cases. Solitary lesions were identified in 18 cases, and carcinoma in situ coexisting with papillary tumours in 36 cases. While PDD identified 92% of the CIS cases, white light cystoscopy identified 68% [7, 8] (Fig. 1). Fig. 1 Image of Cis lesion in white light cystoscopy Image of Cis lesion in PDD mode A disadvantage of PDD is its rather low specificity, ranging from 35% to 66%, which is often due to a lack of experience on the part of the operator or the presence of scars after previous TURBT or intravesical instillation [9]. Nevertheless, PDD is recommended as a diagnostic tool in follow-up examinations of patients with CIS after BCG immunotherapy [10]. In a study of 49 patients with CIS examined after BCG therapy, 18 cases of recurrence were noted. Of these 18 cases, white light cystoscopy revealed no CIS lesions, while PDD diagnosed 14. Overall, while nine of the PDD results were found to be false positives (33.3%), only one positively identified by white light cystoscopy was false (7.1%) [10]. PDD represents a great enhancement of the urological diagnosis of CIS of the urinary bladder and is a superior method to standard white light cystoscopy in cases where CIS is suspected. This efficiency is even more important when considering that a diagnosis of CIS demands the quick implementation of appropriate therapy.

The evaluation of tissue mass loss in the incision line of prostate with benign hyperplasia performed using holmium laser and cutting electrode.

Introduction The aim of this study is to compare the changes in the incision line of prostatic adenoma using a monopolar cutting electrode and holmium laser, as well as the assessment of associated tissue mass and volume loss of benign prostatic hyperplasia (BPH). Material and methods The material used in this study consisted of 74 preparations of prostatic adenoma obtained via open retropubic adenomectomy, with an average volume of 120.7 ml. The material obtained cut in vitro before fixation in formaldehyde. One lobe was cut using holmium laser, the other using a monopolar cutting electrode. After the incision was made, tissue mass and volume loss were evaluated. Thermocoagulation changes in the incision line were examinedunder light microscope. Results In the case of the holmium laser incision, the average tissue mass loss was 1.73 g, tissue volume loss 3.57 ml and the depth of thermocoagulation was 1.17 mm. When the monopolar cutting electrode was used average tissue mass loss was 0.807 g, tissue volume loss 2.48 ml and the depth of thermocoagulation was 0.19 mm. Conclusions Where holmium laser was used, it was observed that the layer of tissue with thermocoagulation changes was deeper than in the case of the monopolar cutting electrode. Moreover, it was noticed that holmium laser caused bigger tissue mass and volume loss than the cutting electrode.

Rare case of malignant lymphoma of kidney

We present a 64–year–old patient suffering from extranodal malignant lymphoma. The patient was admitted to the County Hospital Urology Ward with suspicion of the left kidney tumor. As part of pre–operational diagnosis, ultrasonographic examination and computed tomography of kidneys were conducted. The results confirmed the initial diagnosis. After undergoing radical surgical treatment, the patient is currently under the care of hematology ward of oncology centre.

HIf-1, glut1, endoglin, and BIrC5 expression in urine samples obtained from patients with bladder malignancies - after photodynamic diagnosis (pDD)

Introduction Bladder cancer (BC) is a serious medical problem. The high rate of recurrence and progression demands the development of new methods, such as genetic markers, which allow diagnosis and patient follow-up. Objectives The aim of this study was to compare expression of HIF-1, GLUT1, endoglin, and BRIC5 in patients without and those with BC. The second group was divided into sub-groups: those without a history of PDD (photodynamic diagnosis) in the diagnostic process and those after PDD. Methods Patients with BC were diagnosed using the PDD method using hexaminolevulinate (Hexvix®). The expressions of HIF-1, GLUT1, endoglin, and BRIC5 genes were established in urine specimens by real-time quantitative polymerase chain reaction (PCR). Results The expressions of all tested genes were higher in the group of patients with BC than in the group without BC. In the group after PDD, a statistically significant overexpression of HIF-1 was observed. In this group, changes were not observed in cases of the other three tested genes. Conclusions The differences between the group with PDD and the group without it can be connected with the direct influence of PDD on malignant tissue, which can cause overexpression of HIF-1 only. This is, however, only a hypothesis and needs further study.

Selected examples of complications after minimally invasive treatment for urolithiasis.

In recent years urologists have concentrated on the intense introduction of minimally invasive methods for the treatment of urinary tract diseases with major progress noted in the treatment of urolithiasis. Nowadays extracorporeal shockwave lithotripsy (ESWL), percutaneous nephrolithotomy (PCNL), and ureteroscopic lithotripsy (URSL) are widely used in the treatment of urinary tract lithiasis. The aim of this study is to present examples of urinary tract lithiasis as the complication after minimally invasive methods used in the treatment of urolithiasis. One should remember that even minimally invasive medical procedures using the instruments retained in long-term contact with urine may be the cause of incrustation and stone formation.