Antitumor Efficacy of Bacillus Calmette-Guerin Loaded Cationic Nanoparticles for Intravesical Immunotherapy of Bladder Tumor Induced Rat Model.

For bladder cancer, intravesical chemo/immunotherapy is widely used as adjuvant therapy after surgical transurethral resection. Bacillus Calmette-Guerin (BCG) is a live attenuated Mycobacterium of the same family as tuberculosis, that is capable of inducing a local inflammatory response upon instillation into the bladder. Intravesical therapy with BCG has proved to be more effective in the prophylaxis and treatment of superficial bladder tumors than most chemotherapeutic agents used for the same indication. However, compared to intravesical chemotherapy, BCG immunotherapy provokes more pronounced local and systemic reactions. In addition to the commonly induced granulomatous inflammatory changes in the bladder, which produce irritative symptoms, this therapy may cause systemic side effects varying from mild malaise and fever to, in rare instances, life-threatening or fatal sepsis. Nanoparticles with positive surface charge and mucoadhesive properties were developed to overcome these side effects. Hence, the aim of this study was to optimize and evaluate cationic chitosan (CS) nanoparticles encapsulating BCG in terms of antitumor efficacy after intravesical administration in bladder tumor, induced in rat model. It was found that nanoparticle formulations of 269-375 nm in size can be produced with 42% encapsulation efficiency. The zeta potential was positive and was suitable for intravesical administration. Antitumor efficacy was determined over the parameters of histopathological evaluation, survival rate and mean bladder weight in comparison to treatment with commercial BCG solution. Concerning survival rates, BCG-loaded chitosan nanoparticles resulted in significantly longer survival than BCG commercial product (up to 86 days of survival with no systemic side effects). When compared to healthy bladder weight averages, all groups (especially BCG commercial solution) showed higher bladder weights confirming tumor formation. Histopathological findings confirmed antitumor activity in all treatment groups and optimum findings were observed in groups treated with CS nanoparticles encapsulating BCG. At the same time, significant nanoparticle accumulation in bladder tissues was observed especially for BCG-loaded CS group. In this study, it was clearly observed that cationic CS nanoparticles provide a significantly improved perspective in intravesical immunotherapy of bladder tumors.

Garlic supplemented diet attenuates gentamicin nephrotoxicity in rats.

PURPOSE: To demonstrate the effect of a 4% pulverized garlic supplemented diet on the nephrotoxicity induced by gentamicin in rats. MATERIALS AND METHODS: Twenty four healthy male Wistar rats, weighing between 220 - 260 grams, were divided into three groups. The rats were randomly assigned to either the gentamicin injection without garlic supplementation group (Group I, n = 8), gentamicin injection with garlic supplementation group (Group II, n = 8), and control group (Group III, n = 8). Urine from the rats was collected and the volume (mL), microalbumin (mg/L), creatinine (mg/dL), Na (mmol/L), K (mmol/L), Cl (mmol/L), P (mg/dL), N-acetyl glucosamine (NAG) (U/L) and pH values were measured. Then urea (mg/dL), creatinine (mg/dL), total protein (g/dL) and cystatin (mg/L) values were measured for the blood samples obtained from tail veins. RESULTS: The median NAG value for the control group (52.050 U/L) was similar to value for Group II (56.400 U/L), which received gentamicin and the garlic diet. However, the median NAG value for Group I (77.030 U/L), which received gentamicin without garlic supplementation, was determined to be statistically significantly higher (p = 0.010) than the value for the control group. In addition, the mean cystatin value for Group II (1.360 U/L) was found to be statistically significantly lower than the value for the Group I (2.240 U/L) (p = 0.015). CONCLUSIONS: In this study we showed the effect of 4% pulverized garlic supplemented diet for preventing nephrotoxicity induced by gentamicin in rats by using as parameters NAG in urine samples and cystatin C in serum samples.

Garlic supplemented diet attenuates gentamicin nephrotoxicity ın rats

Purpose To demonstrate the effect of a 4% pulverized garlic supplemented diet on the nephrotoxicity induced by gentamicin in rats. Materials and Methods Twenty four healthy male Wistar rats, weighing between 220 - 260grams, were divided into three groups. The rats were randomly assigned to either the gentamicin injection without garlic supplementation group (Group I, n = 8), gentamicin injection with garlic supplementation group (Group II, n = 8), and control group (Group III, n = 8). Urine from the rats was collected and the volume (mL), microalbumin (mg/L), creatinine (mg/dL), Na (mmol/L), K (mmol/L), Cl (mmol/L), P (mg/dL), N-acetyl glucosamine (NAG) (U/L) and pH values were measured. Then urea (mg/dL), creatinine (mg/dL), total protein (g/dL) and cystatin (mg/L) values were measured for the blood samples obtained from tail veins. Results The median NAG value for the control group (52.050 U/L) was similar to value for Group II (56.400 U/L), which received gentamicin and the garlic diet. However, the median NAG value for Group I (77.030 U/L), which received gentamicin without garlic supplementation, was determined to be statistically significantly higher (p = 0.010) than the value for the control group. In addition, the mean cystatin value for Group II (1.360 U/L) was found to be statistically significantly lower than the value for the Group I (2.240 U/L) (p = 0.015). Conclusions In this study we showed the effect of 4% pulverized garlic supplemented diet for preventing nephrotoxicity induced by gentamicin in rats by using as parameters NAG in urine samples and cystatin C in serum samples. .

Cationic core-shell nanoparticles for intravesical chemotherapy in tumor-induced rat model: safety and efficacy.

Mitomycin C (MMC) has shown potent efficacy against a wide spectrum of cancers and is clinical first choice in superficial bladder tumors. However, intravesical chemotherapy with MMC has been ineffective due to periodical discharge of the bladder and instability of this drug in acidic pH, both resulting in high rate of tumor recurrence and insufficiency to prevent progression. Nanocarriers may be a promising alternative for prolonged, effective and safe intravesical drug delivery due to their favorable size, surface properties and optimum interaction with mucosal layer of the bladder wall. Hence, the aim of this study was to evaluate and optimize cationic core-shell nanoparticles formulations (based on chitosan (CS) and poly-ϵ-caprolactone (PCL)) in terms of antitumor efficacy after intravesical administration in bladder tumor induced rat model. Antitumor efficacy was determined through the parameters of survival rate and nanoparticle penetration into the bladder tissue. Safety of the formulations were evaluated by histopathological evaluation of bladder tissue as well as observation of animals treated with MMC bound to nanoparticles. Results indicated that chitosan coated poly-ϵ-caprolactone (CS-PCL) nanoparticles presented the longest survival rate among all treatment groups as evaluated by Kaplan-Meier plotting. Histopathological evaluation revealed that cationic nanoparticles were localized and accumulated in the bladder tissue. As intravesical chemotherapy is a local therapy, no MMC was quantified in blood after intravesical instillation indicating no systemic uptake for the drug which could have subsequently led to side effects. In conclusion, core-shell type cationic nanoparticles may be effective tools for the intravesical chemotherapy of recurrent bladder tumors.

Prolonged retention and in vivo evaluation of cationic nanoparticles loaded with Mitomycin C designed for intravesical chemotherapy of bladder tumours.

To overcome the recurrence problem in bladder tumours; nanoparticles with positive surface charge may improve interaction with biological membranes for intravesical administration. The aim of this study was to design, develop and evaluate (in vitro-in vivo) cationic nanoparticles based on chitosan, poly-L-lysine or polycaprolactone for the effective intravesical delivery of chemotherapeutic agent MMC in a rat model. Poly-L-lysine-coated polycaprolactone nanoparticles and chitosan-coated polycaprolactone nanoparticles were prepared by the double emulsion technique. Chitosan nanoparticles were prepared by ionic gelation. It was found that nanoparticle formulations of 160-320 nm in size can be produced in 14-35% encapsulation efficiency. Variability in the particle size of nanoparticles depended on the preparation method. Encapsulation was increased by two-fold for CS-PCL as a result of the double emulsion technique. Commercial MMC product in solution form and cationic nanoparticle formulations were compared for in vivo bladder retention properties and effect of formulations on urine volume.

Intravesical cationic nanoparticles of chitosan and polycaprolactone for the delivery of Mitomycin C to bladder tumors.

Cationic nanoparticles of chitosan (CS), poly-epsilon-caprolactone coated with chitosan (CS-PCL) and poly-epsilon-caprolactone coated with poly-L-lysine (PLL-PCL) were developed to encapsulate intravesical chemotherapeutic agent Mitomycin C (MMC) for longer residence time, higher local drug concentration and prevention of drug loss during bladder discharge. Nanoparticle diameters varied between 180 and 340 nm depending on polymer used for preparation and coating. Zeta potential values demonstrated positive charge expected from cationic nanoparticles. MMC encapsulation efficiency depended on hydrophilicity of polymers since MMC is water-soluble. Encapsulation was increased by 2-fold for CS-PCL and 3-fold for PLL-PCL as a consequence of hydrophilic coating. Complete drug release was obtained with only CS-PCL nanoparticles. On the other hand, CS and PLL-PCL nanoparticles did not completely liberate MMC due to strong polymer-drug interactions which were elucidated with DSC studies. As far as cellular interaction was concerned, CS-PCL was the most efficient formulation for uptake of fluorescent markers Nile Red and Rhodamine123 incorporated into nanoparticles. Especially, CS-PCL nanoparticles loaded with Rhodamine123 sharing hydrophilic properties with MMC were selectively incorporated by bladder cancer cell line, but not by normal bladder epithelial cells. CS-PCL nanoparticles seem to be promising for MMC delivery with respect to anticancer efficacy tested against MB49 bladder carcinoma cell line.

Diagnostic value of serum ghrelin levels in prostate cancer.

PURPOSE: Expression of recently identified growth hormone-releasing peptide, ghrelin, and its receptor has been demonstrated in prostate cancer (PCA) cell lines. It was also shown that ghrelin has increased cell proliferation in vitro when added to PCA cell lines. The aim of this study was to evaluate the diagnostic value of serum ghrelin levels in detection of PCA. MATERIAL AND METHOD: 30 patients with PCA and 50 patients with benign prostate hyperplasia (BPH) were enrolled in the study. The serum ghrelin levels of PCA and BPH patients were compared. The correlations between ghrelin and age groups, body mass index, total prostate-specific antigen (PSA) levels, free/total PSA ratio, Gleason score, and prostate volume were also studied. RESULTS: There were no statistically significant differences between the two groups and parameters mentioned above in terms of serum ghrelin levels (p > 0.05). CONCLUSION: Although ghrelin has been shown to induce PCA cell proliferation by in vitro studies, its role in the diagnosis of PCA was not demonstrated in our clinical study. Insufficient secretion of ghrelin into serum or the effect of other sources of ghrelin to serum ghrelin levels could be responsible for this discrepancy.

Values for free/total prostate-specific antigen ratio as a function of age: necessity of reference validation in a Turkish population.

BACKGROUND: The aim of this study was to evaluate age-related changes in free/total prostate-specific antigen (f/t PSA) ratio, focusing on the avoidance of unnecessary prostate biopsies. METHODS: A total of 898 men aged 30-88 years without a history of prostate surgery and disease were enrolled into the study. Serum tPSA, fPSA and f/t PSA ratios were determined for the study population and for different age categories. All males who had suspicious digital rectal examination and tPSA >4 ng/mL underwent transrectal ultrasonography-guided prostate biopsy. Receiver operating characteristic (ROC) curves for each group were generated by plotting the sensitivity vs. 1-specificity for the f/t PSA ratio. The sensitivity and specificity were obtained using different f/t PSA ratio cutoffs for different age groups. RESULTS: Prostate cancer was detected in 63 patients (7%). Age-specific cutoffs were determined according to likelihood ratios at the levels of 10%, 15% and 15% f/t PSA ratio for ages 50-59, 60-69 and >/=70 years, respectively. However, a single cutoff of 10% is recommended across all age ranges (positive likelihood ratio 2.36). ROC curves demonstrated that the area under the curve (AUC) was significant for all patients with initial PSA of 4-10 ng/mL (AUC 0.703-0.796), except for the >/=70-year age group (AUC 0.549). CONCLUSIONS: The current study showed that the use of f/t PSA ratio in patients with PSA levels of 4-10 ng/mL should enhance the specificity of PSA screening and decrease the number of unnecessary biopsies. f/t PSA levels may show dissimilarities according to age and ethnicity, so further studies are warranted to identify this relationship.

Correlations between the ICIQ-SF score and urodynamic findings.

AIMS: The primary aim of this prospective study was to examine the correlations between "The International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF)" score and urodynamic findings in patients with urge incontinence. In addition, we aimed to observe the alterations of these parameters with antimuscarinic therapy. METHODS: Between January and December 2005, patients referred to our department with urge incontinence were examined. After taking a detailed clinical history, physical examination, and urinalysis, each patient was asked to complete an ICIQ-SF questionnaire. We carried out subtracted cystometry according to a fixed protocol on all patients. Patients who were defined as detrusor overactivity incontinent were given antimuscarinic therapy for 3 months. Following treatment, filling cystometry and ICIQ-SF scoring were repeated in all patients. All pre- and post-treatment data of 18 male and 42 female patients were transferred to the SPSS 11.0 for Windows program, and statistical analyses were performed. RESULTS: The patients' ages ranged from 28 to 70 (mean 49.8) years. We found statistically significant differences between the pre- and post-treatment parameters (mean ICIQ-SF score, first sensation, cystometric capacity, maximum detrusor pressure, compliance; P<0.01). We found negative correlation between pre-treatment mean ICIQ-SF score and first sensation (correlation coefficient -0.266, P<0.05) and positive correlation between pre-treatment mean ICIQ-SF score and maximum detrusor pressure (correlation coefficient 0.4, P<0.01). CONCLUSIONS: ICIQ-SF scoring is a practical and reliable method for baseline and post-treatment evaluation of patients with urge incontinence. Significant correlation exists between ICIQ-SF score and urodynamic parameters.

Partial nephrectomy using a monopolar radiofrequency device: an animal model.

OBJECTIVES: To compare the use of radiofrequency (RF) and electrocautery in partial nephrectomy without renal artery clamping for bleeding and tissue destruction. METHODS: Sixteen adult rabbits were randomized into two groups. Partial nephrectomy using a monopolar RF device without renal artery clamping was performed in 8 rabbits (RF group) and partial nephrectomy using electrocautery was performed in the rest (cautery group). Four rabbits in each group (rabbits 1, 3, 5, and 7) were kept for follow-up, and the operated kidneys of the rest were removed for histopathologic evaluation. The tissue samples were placed in 10% formalin solution and sent to the pathology laboratory. The groups were compared in terms of bleeding and tissue destruction. RESULTS: The mean blood loss was 3.6 +/- 1.2 mL in the RF group and 8.3 +/- 2.7 mL in the control group (P = 0.003). A rabbit in the control group died on postoperative day 3 because of bleeding. Others were followed up for 3 months postoperatively. The amount of thermal destruction was comparable between the two groups. Varying degrees of thermal destruction were observed at the cutting margins in both groups. No difference was found between the two groups in terms of the deepness of thermal injury (1 to 2 mm). CONCLUSIONS: The findings of our experimental study showed that the use of the RF electrode in partial nephrectomy without renal artery clamping resulted in less intraoperative bleeding without differences in terms of tissue destruction.

A prospective randomized study for comparing bipolar plasmakinetic resection of the prostate with standard TURP.

OBJECTIVES: To compare clinical results of plasmakinetic (PK) resection vs. standard monopolar resection of the prostate, i.e. transurethral resection of the prostate (TURP). MATERIALS AND METHODS: 48 patients were included in this study between January 2003 and October 2003. They were randomized into two groups (TURP:PK) with a ratio of 1:1. PK resections (n = 24) were carried out by using PlasmaKinetic Tissue Management System (Gyrus Medical Ltd, Cardiff, UK) and PlasmaSect electrodes. TURPs (n = 24) were done by using a 26-Fr continuous-flow resectoscope and Karl Storz 27040 electrodes. Patients were assessed for safety and efficacy by measuring the IPSS and maximum flow rates at 1, 3, 6 and 12 months and residual urine measurement at 3, 6 and 12 months and transrectal ultrasonography at 6 months. RESULTS: The patients' ages ranged from 50 to 82 (mean 64 +/- 10) years. Groups were similar for operation time, bleeding score, resected tissue, catheterization time and irrigated volume. Mean serum Na levels at the end of the operation were 141.7 +/- 5.1 in the TURP group and 145.2 +/- 4.4 in the PK group (p = 0.013). The IPSS, QOL score and Q(max) had improved significantly in the postoperative period without any differences in either group. CONCLUSIONS: The main advantage of PK resection seems to be decreasing the risk of TUR syndrome, thus, larger prostates could be treated without a time limitation, theoretically. However, this technique brings no advantages in terms of intra- and postoperative bleeding, hospital stay, operation time and late complications.

Nocturnal enuresis and allergy.

OBJECTIVE: To investigate whether an interaction exists between nocturnal enuresis and allergy. MATERIAL AND METHODS: Thirty-seven (20 boys, 17 girls) children with monosymptomatic nocturnal enuresis were recruited. We studied an allergy panel that included total IgE, 10 examples of inhalant-specific IgE, 10 examples of food-specific IgE, eosinophilic cationic protein (ECP) and Phadiotop. The same panel was studied in a control group of 18 children without monosymptomatic nocturnal enuresis. RESULTS: We did not determine statistically significant differences between the enuretic group and the control group in terms of levels of total IgE, the 10 examples of inhalant-specific IgE and Phadiotop. However, two (soybean and hazelnut) of the 10 food-specific IgE and ECP levels did differ significantly between the two groups. CONCLUSIONS: This first specific IgE study showed that there may be a relationship between nocturnal enuresis and soybean and hazelnut food allergens. Our findings may explain some cases of nocturnal enuresis. However, further studies are necessary to explain the underlying mechanisms and management of this disorder.