[Prognosis of ureteral dilatation during procedures using a ureteral access sheath in patients with urolithiasis].

AIM: To improve the results of treatment of patients with urolithiasis who underwent endoscopic interventions using a ureteral access sheath (UAS) by developing a predictive model of ureteral dilatation without pre-stenting. MATERIALS AND METHODS: A total of 180 patients with kidney stones up to 20 mm were included in the study. They were divided into two groups: in the group 1 (n=79) UAS of 12/14 Ch was used, while in group II (n=101) UAS of 10/12 Ch was inserted. In group I, 48 (60.8%) patients underwent micropercutaneous nephrolithotomy and in 31 (39.2%) retrograde intrarenal surgery was done, compared to 42 (41.6%) and 59 (58, 4%) of patients in group 2. A non-inclusion criterion was a history of ureteral stenting. At the stage of preoperative diagnosis, 60 minutes before the X-ray examination, the patient took a single dose of 80 mg of furosemide per os to improve visualization of the upper urinary tract. After digital processing of computed tomography data and 3D-reconstruction of the upper urinary tract using the DICOM image processing program "RadiAnt DICOM Viewer," a visual assessment of the ureter was carried out to exclude significant deviations and strictures. The ureteral width was measured at three points: pyeloureteral segment, the level of the iliac bifurcation and intramural part. The number of cases of successful insertion of UAS and the rate of damage to the ureteral wall according to the classification proposed by O. Traxer and A. Thomas (2012) were analyzed. The prediction of successful insertion of a UAS was carried out using ROC analysis. RESULTS: In group 1, successful insertion of UAS was observed in 37 (46.8%) patients compared to 84 (83.2%) patients in group 2. In the remaining 42 (53.2%) and 17 (16.8%) cases, respectively, placement of UAS was not possible due to significant tissue resistance and high risk of traumatic injury. The average ureteral diameter at the points of physiological narrowing in patients with successful insertion of 12/14 Ch UAS were 2.0+/-0.1 mm, compared to 1.2+/-0.4 mm in those with failed insertion (p<0.05). In the group 2, similar indicators were 1.6+/-0.1 mm and 1.2+/-0.5 mm, respectively (p<0.05). According to ROC analysis, the diagnostic efficiency of the predictive model when using 12/14 Ch and 10/12 Ch UAS was confirmed by high AUC values (0.925 [95% CI 0.871-0.98] and 0.944 [95% CI 0.89=0.97], respectively). The total number of patients with ureteral injuries was 35 (44.3%) and 40 (39.6%) in groups with 12/14 Ch and 10/12 Ch UAS, respectively. At the same time, complications of the I degree were observed in 24 (30.4%) patients of the group 1 and in 31 (30.7%) patients of the group 2, while injuries of II degree were detected in 10 (12.7%) and 9 (8.9%) cases, respectively (p>0.05). Only in 1 (1.3%) patient, when 12/14 Ch UAS was inserted, grade III damage to the ureteral wall was determined. CONCLUSION: The proposed technique for measuring the cross-section of the ureter allows to predict the successful insertion of UAS at the preoperative stage. The probability of successful passage of UAS of 10/12 and 12/14 Ch in patients with ureteral diameter in physiological narrowings of more than 1.6 mm and 2 mm, respectively, is 95%. An insertion of UAS is a safe procedure, and most complications are classified as grades I or II.

[The phosphorylation state of transducin beta-subunits].

The supposition that nucleoside diphosphate kinase is the enzyme that phosphorylates transducin beta-subunits on one of the histidine residues (His-266) has been analyzed. It stands the reason that 1) this enzyme is multifunctional and plays in particular the role of protein histidine kinase; and 2) the phosphorylated beta-subunit of transducin may activate transducin via the mechanism of transphosphorylation. Nevertheless, in our experiments, in which different forms of transducin preparations were incubated with α- and ß-isoforms of recombinant rat NDP kinase in the presence of [γ32P]ATP or [γ32P]GTP (specific activity of about 1 Ci/mmol) followed by separation of proteins by electrophoresis and-gel radio-autography, the phosphorylation of the transducin beta-subunit wasn't succeeded to be found. The negative result of our experiments most likely implies that the major part of transducin beta-subunits in the preparations has already been phosphorylated via a process that takes place in vivo.

[Purification of transducin betagamma-subunit complex from isotonic extracts of bovine retinal rod outer segments].

A method for obtaining a free complex of transducin betagamma-subunits from bovine retinal rod outer segments in a highly purified state has been suggested.

[Nucleoside diphosphate kinase and GTP-binding proteins. Possible mechanisms of coupling].

The results of numerous investigations during the last 20 years have shown that nucleoside diphosphate kinase (NDP kinase) is a multifunctional protein. In this paper, the current data are analyzed indicating that one of the possible mechanisms by which NDP kinase manifests its multifunctional role is its participation in the activation (or regulation) of heterotrimeric GTP-binding proteins (G proteins). We demonstrate that one of the NDP kinase isoforms dynamically interacts with the retinal rod G protein transducin (Gt) and phosphorylates its beta-subunit at histidine residue (His 266). It is also shown that it leads to the consecutive transfer of the phosphate group to the GDP in the active center of G protein alpha-subunit and G protein activation. The advantages of this mechanism are considered as compared to the classic G protein activation mechanism, GDP/GTP exchange.

Investigation of chimerical and tagged forms of recombinant rat nucleoside diphosphate kinases alpha and beta. Interaction with rhodopsin-transducin complex and thermal stability.

To elucidate the physicochemical basis of differences between the isoforms of mammalian multifunctional nucleoside diphosphate kinase (NDP), we investigated the recombinant rat homohexameric NDP kinases alpha and beta, consisting of highly homologous alpha or beta subunits of 152 residues each and differing only in variable regions V1 and V2, and their chimerical forms (NDP kinase alpha(1-130)beta(131-152) and NDP kinase beta(1-130)alpha(131-152)) and tagged derivatives (NDP kinase HA-alpha(1-130)beta(131-152), NDP kinase HA-beta(1-130)alpha(131-152), and NDP kinase HA-beta). The thermal stability of these proteins and the ability of some of them to interact with the rhodopsin-transducin (R*Gt) complex have been studied. It was found that NDP kinase alpha, NDP kinase alpha(1-130)beta(131-152), and NDP kinase HA-alpha(1-130)beta(131-152) were similar in their thermal stability (T(1/2) = 61-63 degrees C). NDP kinase beta, NDP kinase beta(1-130)alpha(131-152), NDP kinase HA-beta(1-130)alpha(131-152), and NDP kinase HA-beta were inactivated at a lower temperature (T(1/2) = 51-54 degrees C). NDP kinase HA-alpha(1-130)beta(131-152) interacted with the R*Gt complex in the same manner as NDP kinase alpha, whereas the interaction of NDP kinase HA-beta(1-130)alpha(131-152) and NDP kinase beta with the photoreceptor membranes under the same conditions was very weak. It is suggested that the variability of the region V1 is a structural basis for the multifunctionality of NDP kinase hexamers in the cell.