[Results of percutaneous nephrolithotomy using two accesses in the treatment of patients with staghorn stones].

INTRODUCTION: In order to improve clinical efficiency and reduce the risk of postoperative complications in patients with staghorn stones, we compared the results of original technique of biportal percutaneous nephrolithotomy (PCNL) with the standard PCNL. MATERIALS AND METHODS: The total of 221 patients with staghorn stones of K3-K4 was included in the study. The biportal PCNL was used in 109 patients, while the control group consisted of 112 patients. Inclusion criteria were stone size more or equal 2 cm, age over 18 years, absence of coagulopathy and width of the renal parenchyma more or equal 1 cm. On 1st postoperative day, ultrasound or plain urography was performed, while in patient with radiolucent stones, multi-slice computed tomography was used. In addition, complete blood count and biochemical profile were done. The main difference from the standard PCNL with sequential renal tracts is the simultaneous creation of the main and additional accesses when performing biportal PCNL. This method allowed two surgeons to simultaneously and synergistically perform lithotripsy and stone extraction from two accesses using a standard nephroscope in the main tract of 24 Ch and a miniaturized nephroscope in the additional tract of 16.5 Ch. RESULTS: The stone-free rate in the group of biportal PCNL was 80.7% (n=88), compared to 72.3% in the control group (n=81). Secondary interventions and additional procedures were required in 29 (26.6%) and 40 (39.2%) cases, respectively. The total number of infectious and hemorrhagic complications was higher in the control group. DISCUSSION: According to our data, significant advantages are observed in the group of biportal PCNL compared to the standard technique. CONCLUSION: Biportal PCNL can be recommended as a promising advancement of the technique traditionally used in the clinical practice.

[Bacteriophage Therapy of Urological Infections].

A literature review dedicated to the problem of bacteriophage therapy for infectious and inflammatory diseases, including urological infections, is presented. Considering the growth of antibiotic resistance, the search for alternative treatments is relevant. Current data about bacteriophages and mechanism of their action, difference of virulent and temperate phages is given, as well as mechanisms of bacteria resistance to phages and ways of its overcoming. The history of phage therapy for infectious diseases from the beginning of the 20th century to the present days is presented. Pharmacokinetic studies of phages after oral administration are given. Moreover, we described our 30 years experience on of clinical use of bacteriophage cocktails in the treatment and prevention of urological infection. In addition, problems of phage therapy are discussed, including immunological issues and the advantages of bacteriophages over antibiotics.

[Prevention of infectious and inflammatory complications after percutaneous nephrolithotomy].

INTRODUCTION: Given the increase in antibiotic resistance of uropathogens, one of the urgent problems is a development of optimal antimicrobial prophylaxis for surgical interventions, as well as an adequate regimen of antibiotic therapy after percutaneous nephrolithotomy (PCNL). AIM: to determine an effective perioperative antimicrobial prophylaxis for PCNL in patients with kidney stones. MATERIAL AND METHODS: A total of 90 patients with staghorn or multiple large kidney stones who underwent PCNL were included in the study. Before PCNL, urine culture was performed in all patients in order to determine the sensitivity not only to antibiotics, but also to bacteriophages. In addition, urine was taken for additional microbiological studies after the puncture of the collecting system, as well as on the 3rd and 7th day after PCNL. All patients were divided into three groups of 30 patients depending on the regimen of perioperative prophylaxis. In group 1, patients were prescribed ciprofloxacin 1000 mg i.v. intraoperatively, then 1000 mg i.v. q.d. for 3-5 days. In the group 2, patients received one dose of cefotaxime + sulbactam (1.0 g + 0.5 g) 2 hours before PCNL i.m. In the group 3, a polyvalent pyobacteriophage purified was given orally 1 hour before PCNL in a dose of 40 ml and the same dose was used t.i.d. for 3-5 days postoperatively. RESULTS: In all three groups of patients, the following infectious complications were evaluated: acute pyelonephritis, systemic inflammatory response syndrome (SIRS) and urosepsis. There were no serious infectious and inflammatory complications in the early postoperative period among all patients. SIRS developed on days 1-3 after PCNL in 26.6%, 20% and 20% of patients in group 1, 2 and 3, respectively. However, by days 4-7 after PCNL, there was normalization of blood cells count (leukocytes, neutrophil band cells), temperature and general condition. CONCLUSION: Different regimens of antimicrobial prophylaxis for PCNL have the same efficiency. The development of SIRS on days 1-3 after PCNL is correlated not only with the antimicrobial agents used and the route of their administration (intravenously, intramuscularly and orally). Most likely, the development of SIRS is more associated with surgical trauma.

[Effectiveness of laparoscopic repair of primary strictures of ureteropelvic junction].

AIM: To evaluate the effectiveness of laparoscopic repair of primary strictures of ureteropelvic junction (UPJ) depending on baseline renal function of the ipsilateral kidney. MATERIALS AND METHODS: The study analyzed results of 134 patients (78 women and 56 men, age from 18 to 56 years) who underwent various types of laparoscopic repair of the UPJ stricture from 2012 to 2015. Depending on the surgical technique all patients were divided into three groups: group 1 (n=34) underwent spiral flap technique by Culp and DeWeerd, group 2 (n=59) - Anderson-Hynes pyeloplasty and group 3 (n=41) had antevasal dismembered pyeloplasty. All interventions ended with internal ureteral stenting for up to 6-8 weeks. Also, all patients were divided into three subgroups, depending on the degree of renal function deficiency - less than 25%, 25-50%, and 51-75%. Treatment effectiveness criteria included the following parameters: complete relief of the pain syndrome, a decrease in the degree of pyeloectasia, stabilization or improvement of the functional state of the renal parenchyma (according to radioisotope renography), and the absence of recurrence of the UPJ stricture. RESULTS: The overall effectiveness of UPJ laparoscopic reconstruction was 94.7% (127 of 134). The effectiveness of the treatment was independent of the surgical technique, the initial thickness of the renal parenchyma and the degree of PCS dilatation. There was an inverse correlation between the treatment effectiveness the degree of kidney function deficiency. CONCLUSION: In patients with hydronephrosis secondary to UPJ stricture, the effectiveness of surgical treatment is mainly determined by its timeliness. The best treatment results were observed in patients with better renal function. The degree of renal function deficiency should be considered the main prognostic factor for the effectiveness of the forthcoming operation.

Inhibition of GSK-3ß decreases the ischemia-induced death of renal cells.

Pharmacological preconditioning with insulin and lithium ions prevented the death of renal cells under conditions of ischemia/reperfusion. Preincubation of cells with insulin or lithium ions decreased production of reactive oxygen species after ischemia/reoxygenation. These agents also prevented the development of mitochondrial dysfunction in renal cells induced by ischemia/reoxygenation. It was hypothesized that the protective effects of these agents are related to inhibition of glycogen synthase kinase-3(. This enzyme is inactivated upon phosphorylation of serine residue in position 9. We found that in vivo administration of lithium ions to animals before renal ischemia prevents the development of kidney failure.

[Mechanism of action and efficacy of litium chloride in thermal ischemia of the kidney].

A protective effect of lithium chloride (LC) in thermal ischemia of the kidney of different duration was studied in rats. LC efficacy was estimated by functional activity of ischemic kidney in early and late reperfusion period, by damage to mitochondria of tubular epithelial cells and production of active oxygen forms (AOF) and nitric oxide (NO). LC has a marked anti-ischemic effect. In thermal renal ischemia for 40 and 60 min LC provides functional safety of the ischemic organ. In longer ischemia, 50% of the animals died. The mechanism of the protective action of LC is related to reduction of APK production, support of a high transmembrane potential of mitochondria and NO synthesis redistribution in the cells of tubular epithelium cells.

Morphological changes in the kidneys of rats with postischemic acute renal failure after intrarenal administration of fetal mesenchymal stem cells from human bone marrow.

Chronic experiments on outbred albino rats were performed to compare the dynamics of histological signs for postischemic renal injury (90-min thermal ischemia) after intraparenchymal injection of cultured fetal MSC from human bone marrow. Functional indexes of the ischemic kidney were predetermined. In the early period after ischemia (day 4), administration of human bone marrow MSC was followed by the increase in blood flow in the microcirculatory bed and decrease in the degree of alteration in renal tubules. An increase in the area of zones with histological signs for normal function of tubules was accompanied by the improvement of biochemical indexes for renal function. In the delayed period, a protective effect of cell therapy was manifested in the prevention of death of renal tubules. Mild calcification of the necrotic tubular epithelium served as a marker of this process. Human bone marrow MSC were labeled with the fluorescent probe Calcein. These cells migrated from the site of injection, spread in the interstitium, and retained viability for 7 days. During this period, some cells were incorporated into the lumen of renal tubules.

[Clinicobiochemical study of diagnostic value of the panel including 10 potential protein markers of prostatic cancer].

We tested diagnostic value of the panel of 10 specially selected proteins--potential markers of prostatic cancer. A double blind method and proteomic technologies were used in complex clinicobiochemical examination of 20 patients with benign and malignant tumors. The same diagnosis were obtained by clinicomorphological criteria and protein markers in 13 (65%) cases. The highest diagnostic efficacy was achieved in prostatic cancer--11 cases (79%) vs 14 by clinicomorphological criteria.

Mitochondria-targeted plastoquinone derivatives as tools to interrupt execution of the aging program. 2. Treatment of some ROS- and age-related diseases (heart arrhythmia, heart infarctions, kidney ischemia, and stroke).

Effects of 10-(6'-plastoquinonyl) decyltriphenylphosphonium (SkQ1) and 10-(6'-plastoquinonyl) decylrhodamine 19 (SkQR1) on rat models of H2O2- and ischemia-induced heart arrhythmia, heart infarction, kidney ischemia, and stroke have been studied ex vivo and in vivo. In all the models listed, SkQ1 and/or SkQR1 showed pronounced protective effect. Supplementation of food with extremely low SkQ1 amount (down to 0.02 nmol SkQ1/kg per day for 3 weeks) was found to abolish the steady heart arrhythmia caused by perfusion of isolated rat heart with H2O2 or by ischemia/reperfusion. Higher SkQ1 (125-250 nmol/kg per day for 2-3 weeks) was found to decrease the heart infarction region induced by an in vivo ischemia/reperfusion and lowered the blood levels of lactate dehydrogenase and creatine kinase increasing as a result of ischemia/reperfusion. In single-kidney rats, ischemia/reperfusion of the kidney was shown to kill the majority of the animals in 2-4 days, whereas one injection of SkQ1 or SkQR1 (1 micromol/kg a day before ischemia) saved lives of almost all treated rats. Effect of SkQR1 was accompanied by decrease in ROS (reactive oxygen species) level in kidney cells as well as by partial or complete normalization of blood creatinine and of some other kidney-controlled parameters. On the other hand, this amount of SkQ1 (a SkQ derivative of lower membrane-penetrating ability than SkQR1) saved the life but failed to normalize ROS and creatinine levels. Such an effect indicates that death under conditions of partial kidney dysfunction is mediated by an organ of vital importance other than kidney, the organ in question being an SkQ1 target. In a model of compression brain ischemia/reperfusion, a single intraperitoneal injection of SkQR1 to a rat (1 micromol/kg a day before operation) effectively decreased the damaged brain area. SkQ1 was ineffective, most probably due to lower permeability of the blood-brain barrier to this compound.

Effects of ischemic and hypoxic preconditioning on the state of mitochondria and function of ischemic kidneys.

Laser confocal microscopy showed that fluorescence of tetramethylrhodamine ethyl ether probe specifically accumulating in energized mitochondria significantly decreased in renal tubular epithelium after 40-min thermal ischemia, while fluorescence of dichlorodihydrofluorescein and diaminofluorescein probes in the same structures increased under these conditions, which attests to increased generation of ROS and NO, respectively. These forms were generated predominantly in mitochondria of tubular epitheliocytes. Hypoxic preconditioning (a series of sessions of breathing hypoxic mixture) preserved functional activity of mitochondria and prevented activation of ROS and NO generation. Ischemic preconditioning of the kidney consisting of three preliminary episodes of vascular clamping (5 min with 5 min reperfusion periods) also increased the percentage of functionally active mitochondria and prevented activation of NO synthesis without appreciably modifying ROS production. Both protective methods significantly reduced the severity of postischemic dysfunction of the kidney.

Experimental intravenous cell therapy of acute and chronic renal failure.

The therapeutic effect of intravenous injection of human fetal bone marrow mesenchymal stem cells or summary culture of kidney cells were studied on models of chronic or acute renal failure in outbred albino rats. Both cell types promoted improvement and normalization of the renal function in rats with stable chronic renal insufficiency (2 weeks after kidney cell injection, 1 month after bone marrow mesenchymal stem cell injection). Renal function remained normal or subnormal during the delayed period (3-3.5 months after injection). In rats with latent stage of chronic renal insufficiency, exacerbation was induced by additional 40-min ischemia. All rats receiving intravenous injection of saline died. Improvement of the functional parameters started 2 weeks after injection of kidney cells or bone marrow mesenchymal stem cells, and normalization was observed after 1.1-5 months. During the delayed period (after 3-4 months), functional parameters retained at normal or subnormal levels. In experimental series III, all rats with acute renal failure intravenously injected with saline (control) died from uremia on days 2-4. After injection of kidney cells 50% rats survived and renal function in these animals returned to normal after 2 weeks. After injection of bone marrow mesenchymal stem cells 83% rats survived, functional parameters returned to normal after 3 weeks.

The role of mitochondria in oxidative and nitrosative stress during ischemia/reperfusion in the rat kidney.

Reoxygenation following ischemia causes tissue oxidative stress. We studied the role of oxidative stress caused by kidney ischemia/reperfusion (I/R) on the mitochondria of renal tissue slices. I/R caused the mitochondria to be swollen, fragmented, and have lower membrane potential. The mitochondria generated more reactive oxygen species (ROS) and nitric oxide (NO) in situ as measured by fluorescence of ROS- and NO-sensitive probes. Infusion of lithium ion, an inhibitor of glycogen kinase synthase-3, caused phosphorylation of its Ser-9 and restored the membrane potential and decreased ROS production of the mitochondrial fraction. Ischemic kidney and hypoxic rat preconditioning improved mitochondrial membrane potential and lowered ROS production caused by subsequent I/R similar to lithium ion infusion. Preconditioning normalized NO production in mitochondria as well. The drop in the mitochondrial membrane potential was prevented by NO synthase inhibition, demonstrating a strong contribution of NO to changes in mitochondrial energy metabolism during the I/R transition. Mitochondria in the I/R-stressed kidney contained less cytochrome c and more pro-apoptotic Bax, consistent with apoptotic degradation.

[Application of cell technologies for therapy of chronic renal insufficiency (experimental study)].

The experiments on 29 white non-inbred rats with chronic renal failure (CRF) induced by right-side nephrectomy and coagulation of 1/2-2/3 of parenchyma of the left kidney were made to study the trend in renal function after injection (into renal cortex or intravenously) of cultured stem or progenitor cells from human fetuses (total culture of fetal kidney or mesenchymal stem cells of the bone marrow). In control tests with salt solution functional indices reflected persistence of CRF. On day 4 after introduction of the fetal cells into renal parenchyma renal function improved and normalized in 2 weeks. After intravenous injection of fetal cells CRF reduced slowly, especially after injection of medullary mesenchymal cells with normalization in 1 month. 2.5-3.5 months after the injection test parameters in some rats deteriorated but remained close to normal values. Glomerular filtration after injection of stem and progenitor cells recovered better while canalicular sodium reabsorption underwent normalization but was followed by deterioration.

Functional aftereffects of intraparenchymatous injection of human fetal stem and progenitor cells to rats with chronic and acute renal failure.

Chronic renal insufficiency was modeled in rats by unilateral nephrectomy and electrocoagulation of both poles of the remaining kidney; acute renal failure was induced by 90-min clamping of the vascular pedicle of the only kidney. Injection of unfractionated culture of human fetal kidney cells or bone marrow mesenchymal stem cells into damaged kidney restored its function in rats with chronic renal insufficiency (observation period up to 2 months). After 2.5 months a relapse of chronic renal insufficiency was observed in 1 of 3 rats receiving human fetal kidney cells and in 1 of 2 animals receiving bone marrow mesenchymal stem cell culture. Injection of bone marrow mesenchymal stem cell culture to rats with acute renal failure improved recovery of renal function and prevented the death from uremia, while injection of total culture of human fetal kidney cells had virtually no effect on the course of acute renal failure.

[The role of the mitochondria generating reactive oxygen species and nitric oxide in postischemic functional disturbance of the kidney].

Experiments on 10 rats and 10 rabbits were made to investigate metabolic aftereffects of 40-minute heat ischemia and subsequent reperfusion. It was found that mitochondrial function deteriorated significantly in an early postischemic period. The disorder manifested with a relative prevalence of cell ATP consumption over its synthesis. This is accompanied with intensive production by mitochondria of nitric oxide and oxygen free radicals. Fluorescent probes and confocal microscopy of vital renal sections showed that mitochondria are responsible for excessive generation of nitric oxide and oxygen radicals in the kidney in an early reperfusion period. The discussion concerns the role of nitric oxide in reperfusion renal damage and participation of mitochondria in formation of its anti-ischemic resistance.

[Identification of AGR2 protein, a novel potential cancer marker, using proteomics technologies].

A comparative analysis of the proteins in prostate tissues of the patients operated for hyperplasia (n = 7) or cancer (n = 5) was performed aiming to search for protein diagnostic markers. Differences in several minor proteins were detected using two-dimensional electrophoresis according to O'Farrel; among them, an additional protein with a molecular weight of 19 kDa and an isoelectric point of 9.0 was observed in four of the cancer cases. Mass spectrometry allowed this protein to be identified as the androgen-induced secreted protein AGR2. The possibility of using AGR2 as a diagnostic marker of prostate cancer is discussed.

[Quality of life of elderly patients with nephrolithiasis].

Clinical findings have been analysed for 105 presenile and senile patients with nephrolithiasis. A total of 168 operations have been made: open intervention (n=22), percutaneous nephrolitholapaxia (n=12), extracorporeal lithotripsy (n=134). Quality of life was assessed according to the questionnaire survey (an original visual graphic questionnaire on the patient's attitude to his/her disease and the treatment, questionnaire on the doses of analgetics and antibiotics, conventional questionnaire EQ-SD. It was found that elderly patients note a statistically significant fall in frequency of pains (renal colics, lumbar pains, subfebrile condition), in the dose of analgetics and antibacterial drugs 6-12 months after removal of the concrement. Removal of the concrement had a positive influence on quality of life of patients with urolithiasis. Extracorporeal lithotripsy had a weaker emotional effect on the patients than open surgical interventions. It is suggested to remove nephroliths if contraindications are absent.

[New approaches to molecular diagnosis of prostatic cancer].

We compared prostatic proteins in patients operated for adenoma or cancer. 630 protein fractions were obtained from each tissue sample after fractionation by two-dimentional electrophoresis according to O'Farrell. Comparison of the samples from adenoma and cancer showed their difference by 7 proteins among which were isoforms of glyceraldehydes-3-phosphate-dehydrogenase, alpha-collagen and several little known proteins. Most of the cancer patients had the protein with molecular mass 19 kDa and isoelectric point 9.0. By the results of mass-spectrometry this protein was identified as androgen-induced secreted protein AGR2. This protein is considered a potential oncomarker. Prospects of some postgenome technologies for detection of new diagnostic markers of prostatic cancer are discussed.