[Application of laser technologies in the treatment of non-muscle-invasive bladder cancer].

This review includes the facts of bladder treatment by means of laser technologies application in historical perspective. Advantages and disadvantages of different laser technologies were considered in comparison with the traditional TUR of the bladder wall.

[Focal laser coagulation in the treatment of a patient with kidney cancer and prostate cancer].

The article describes the clinical observation of the patient 84 years old with prostate cancer T2bNxM0, left kidney cancer T1N0M0 and severe comorbidity. Stepwise interstitial laser coagulation was performed using the Russian microsecond Nd: YAG laser surgical complex.

[Age-related androgen deficiency. The current state of the problem].

This paper reviews the modern concept of age-related androgen deficiency. The authors discuss the prevalence of androgen deficiency and its relationship with age, concomitant diseases, pharmacotherapy and other factors.

[Large urethral stones: a case report].

The article describes a clinical observation of a 68 y/o patient with large stones of the urethra, reaching 5 cm in length and located in the penile urethra. The patient underwent a three-stage treatment: the first stage consisted of drainage of the urinary bladder, the second stage of the urethrolithotomy and a double-barreled fistula, the third was an anastomotic urethroplasty. After removing the cystostomic drainage tube, normal urination was restored.

[EXPERIMENTAL VALIDATION OF ND:YAG LASER COAGULATION OF THE BLADDER TISSUE].

Interstitial laser coagulation (ILC) is one of the minimally invasive and minimally traumatic methods for the destruction of small lesions in various organs and tissues. Selection of the optimal operation and exposure mode is critical for the safe use of ILC in clinical practice. The in-vivo study of ILC effects on bladder tissue was carried out in 6 rabbits weighing less than 3 kg. Macroscopic and microscopic analysis of the tissue was done directly after the ILK procedure, on days 3 and 7, 14 and 21 following specimen collection of experimental animals. Safety and efficacy of ND: YAG laser impact on bladder tissues were confirmed by morphological study, which established phasing and extent of changes in the bladder tissues. The results allow us to recommend ILC application for the destruction of small size bladder tumors without impairment of the bladder functions, as well as for the treatment of bottom after transurethral resection of bladder cancer.

Purification of high-molecular-weight subfraction from porcine skin inhibiting proliferation of A431 human carcinoma epidermoid cells.

Subfraction with a molecular weight >250 kDa isolated from porcine skin and inhibiting the proliferation of A431 human carcinoma epidermoid cells was purified by DEAE 32 anion exchange chromatography with NaCl concentration step-gradient. The effects of the initial subfraction and fractions obtained by separation in DEAE 32 on the proliferation of A431 human carcinoma epidermoid cells were studied in vitro in two tests (MTT and fluorescent test). The more sensitive fluorescent test showed the highest inhibitory activity of fraction No. 2 released from the column at 0.15 M NaCl. One major protein component and a series of minor protein components were detected in this fraction by vertical PAAG-SDS electrophoresis.

[Features of endoscopic surgical treatment of urolithiasis in overweight patients].

The article presents the results of endoscopic and nephrolithotripsy and ureterolithotripsy in 73 overweight patients (body mass index of over 30 kg/m2) with kidney stones and urinary tract infections of different locations compared with 30 patients without obesity. The features of choice of tactics of surgical treatment and access, possible complications and their prevention are described.

[Modern tactics of management of patients with bilateral staghorn nephrolithiasis].

A comparative analysis of the effectiveness of open and percutaneous operations in the treatment of bilateral staghorn nephrolithiasis (SN) was performed. Of the 38 patients with bilateral SN, stage operation (without discharge) was performed in 26 (68.5%) patients, and single-step operation--in 12 (31.5%) patients. For the 47 patients with bilateral SN, who underwent open surgery, the interventions were following: pelviolithotomy (64 operations), nephrolithotomy (21 operations), nephrectomy (9 operations). Surgery was performed in stages on each side, but unlike percutaneous operations, with discharge from the hospital. When using open surgery, only in 27 of the 47 patients complete discharge of kidney stones was achieved. A comparative analysis of single-step and stage surgery for bilateral SN revealed no significant difference in the duration of surgery and intraoperative blood loss (p > 0.1); endoscopic surgery compared with open surgery is characterized by significantly lower intraoperative blood loss and postoperative in-hospital stay (p < 0.001).

[Experimental validation of the use of ND:YAG-interstitial laser coagulation of the kidney].

Interstitial laser coagulation (ILC) is one of the effective methods of minimally invasive destruction of small renal tumors. For the safe use of ILC in clinical practice, it is necessary to select the optimal mode of action of laser radiation on the tissue of target organ and techniques for the procedure. The effects of ILC on kidney tissue in 6 rabbits weighing less than 3 kg and 5 mongrel dogs weighing up to 20 kg were investigated in vivo. The impact of ILC was made intraoperatively. Organ retrieval in experimental animals for the macro- and microscopic examination was performed immediately after the impact of ILC and at various times thereafter during the month in rabbits and 35 days in dogs. Morphological study in different periods after exposure allowed to establish phasing and volumes of changes in the tissues of the kidney, confirming the safety and efficacy of ND:YAG laser irradiation on kidney tissues. The results indicate the possibility of effective application of ILC of kidney tissues for destruction of small size tumors, without impairment of functions of the organ.

[Assessment of morphological changes in oral mucosa in patients with gastrointestinal pathology].

Anatomical and functional proximity of oral and digestive tract organs as well as common mechanism of innervation and humoral regulation create prerequisites for the involvement of oral cavity in pathological processes developing in the gastrointestinal tract. Various gastric and duodenal substances affect oral tissues during frequent refluxes and produce lesions in epithelium of oral mucosa. Results of preliminary qualitative assessment of morphological changes in lingual mucosa are presented for patients with acid-dependent pathology.

Molecular design of new inhibitors of peroxidase activity of cytochrome c/cardiolipin complexes: fluorescent oxadiazole-derivatized cardiolipin.

Interaction of a mitochondria-specific anionic phospholipid, cardiolipin (CL), with an intermembrane protein, cytochrome c (cyt c), yields a peroxidase complex. During apoptosis, the complex induces accumulation of CL oxidation products that are essential for detachment of cyt c from the mitochondrial membrane, induction of permeability transition, and release of proapoptotic factors into the cytosol. Therefore, suppression of the peroxidase activity and prevention of CL oxidation may lead to discovery of new antiapoptotic drugs. Here, we report a new approach to regulate the cyt c peroxidase activity by using modified CL with an oxidizable and fluorescent 7-nitro-2,1,3-benzoxadiazole (NBD) moiety (NBD-CL). We demonstrate that NBD-CL forms high-affinity complexes with cyt c and blocks cyt c-catalyzed oxidation of several peroxidase substrates, cyt c self-oxidation, and, most importantly, inhibits cyt c-dependent oxidation of polyunsaturated tetralinoleoyl CL (TLCL) and accumulation of TLCL hydroperoxides. Electrospray ionization mass spectrometry and fluorescence analysis revealed that oxidation and cleavage of the NBD moiety of NBD-CL underlie the inhibition mechanism. We conclude that modified CL combining a nonoxidizable monounsaturated trioleoyl CL with a C(12)-NBD fragment undergoes a regiospecific oxidation thereby representing a novel inhibitor of cyt c peroxidase activity.

Bifunctional anti/prooxidant potential of metallothionenin: redox signaling of copper binding and release.

Metallothioneins (MTs) are cysteine-rich metal-binding proteins that exert cytoprotection during metal exposure and oxidative stress. The roles of MT in copper (Cu) binding and release and modulation of redox cycling are unresolved. We hypothesized that Cu-binding to MT renders Cu redox inactive, but that oxidation of free thiols critical for metal binding can reduce MT/Cu interactions and potentiate Cu redox cycling. Overexpression of MT in cells by cadmium pretreatment or ectopic overexpression by gene transfer confers protection from Cu-dependent lipid oxidation and cytotoxicity. Using a chemically defined model system (Cu/ascorbate/H2O2) to study Cu/MT interactions, we observed that MT inhibited Cu-dependent oxidation of luminol. In the absence of H2O2, MT blocked Cu-dependent ascorbyl radical production with a stoichiometry corresponding to Cu/MT ratios < or = 12. In the presence of H2O2, Cu-dependent hydroxyl radical formation was inhibited only up to Cu/MT ratios < or = 6. Using low-temperature EPR of free Cu2+ to assess Cu/MT physical interactions, we observed that the maximal amount of Cu1+ bound to MT corresponded to 12 molar equivalents of Cu/MT with Cu and ascorbate alone and was reduced in the presence of H2O2. 2,2'-Dithiodipyridine titration of MT SH-groups revealed a 50% decrease after H2O2, which could be regenerated by dihydrolipoic acid (DHLA). DHLA regeneration of thiols in MT was accompanied by restoration of MT's ability to inhibit Cu-dependent oxidation of ascorbate. Thus, optimum ability of MT to inhibit Cu-redox cycling directly correlates with its ability to bind Cu. Some of this Cu, however, appears releasable following oxidation of the thiolate metal-binding clusters. We speculate that redox-dependent release of Cu from MT serves both as a mechanism for physiological delivery of Cu to specific target proteins, as well as potentiation of cellular damage during oxidative stress.

Mishandling of copper by albumin: role in redox-cycling and oxidative stress in preeclampsia plasma.

OBJECTIVE: To test the hypothesis that enhanced oxidative stress during pregnancies complicated by preeclampsia is associated with improper copper (Cu) binding by plasma albumin, resulting in enhanced Cu redox-cycling activity and that altered Cu binding, in turn, is caused by interactions of excessive amounts of free fatty acids with albumin. STUDY DESIGN: We studied binding and redox-cycling activity of Cu in 17 normal pregnancy and 17 preeclampsia plasma samples. Binding of exogenous Cu in plasma samples was quantified indirectly using spectrophotometric measurements of its complex with a specific chelator of Cu(I), bathocuproine disulfonate. Redox-cycling activity of Cu in plasma samples was estimated by electron paramagnetic resonance (EPR) spectroscopy of ascorbate radicals formed during one-electron oxidation of ascorbate by redox-active catalytic Cu. Formation of Cu/albumin complexes in model systems in the presence and absence of fatty acids was studied using EPR spectroscopy of Cu(II)/albumin. RESULTS: We found that preeclampsia plasma (as compared to normal pregnancy plasma) (1) displays elevated endogenous ascorbate redox-cycling that is normalized by a Cu(II) chelator, cuprizone I, (2) has lowered capacity to bind and redox-regulate exogenously added Cu, and (3) responds to treatment with fatty-acid-free albumin by diminished ascorbate oxidizing activity. Conversely, addition of free fatty acid (oleic acid) to normal pregnancy plasma sample yields increased ascorbate redox-cycling activity. Our model experiments showed that Cu-dependent redox-cycling activity of purified human serum albumin is significantly increased by excess free fatty acids. CONCLUSION: Mishandling of Cu by albumin contributes to oxidative stress in preeclampsia. Cu chelators may represent promising mechanism-based antioxidants to attenuate oxidative stress in preeclampsia.

[Molecular and cellular mechanisms of the low intensity laser radiation effect]. / Molekuliarno-kletochnye mekhanizmy deistviia nizkointensivnogo lazernogo izlucheniia.

The main aspects of the free radical conception of the molecular and cellular mechanisms of the stimulating action of low-intensity radiation in the red region of the spectrum were considered. These are: (1) Primary acceptors of incident radiation are endogenous porphyrins, which may act as photosensitizers giving initiator-radicals for secondary free radical reactions. (2) Target cells for light irradiation during quantum therapy may be blood leukocytes, fibroblasts, keratinocytes, endotheliocytes, etc. (3) The initiation of the secondary free radical reactions due to lipid peroxidation of cell membranes (in particular, of leukocytes) brings about an increase in ion permeability including that for calcium. The increase in intracellular calcium concentration leads to phagocytes priming, i.e., to increased production of reactive oxygen species (ROS) under subsequent stimulation of the cell. (4) Photosensitized generation of ROS in the cytoplasm of some cells induces a free-radical activation of synthesis of proteins, the most significant in the light of the present concept being the de novo synthesis of inducible NO-synthase, superoxide dismutase, and various cytokines. The experimental evidence for the basic statements of the conception of free radical mechanisms for the stimulating action of low-intensity laser and noncoherent radiations is presented. A relation between the primary mechanisms of the stimulating action of light and the secondary effects that determine the sanative effect of quantum therapy in the process of wound healing (bactericidity, cell proliferation, and improved microcirculation) was established. Moreover, it was shown that nitrosyl complexes of heme proteins, such as hemoglobin and cytochrome c, are the primary chromophores of laser radiation. Upon irradiation, they can easily dissociate to produce free nitric oxide. In turn, released nitric oxide may be responsible for blood vessel relaxation and activation of mitochondrial respiration. This phenomenon is just observed during phototherapy by means of low-intensity laser radiation.

[Nitric oxide, hemoglobin and laser irradiation]. / Oksid azota, gemoglobin i lazernoe obluchenie.

The paper deals with the present views of some chemical properties and biological effects of nitric oxide (NO) and chiefly its formation and primary conversions. The interaction of NO with superoxide radical is shown to one of the most important reactions of the former, which gives rise to peroxynitrite whose breakdown yields a hydroxy radical. Emphasis is laid on the substances that are a temporary depot or a possible transport form of NO, such as nitrosothiols and nitrosyl complexes of non-hemic iron. NO is well-known to release when these compounds are degraded. It is suggested that hemoglobin is another NO depot, which forms stable complexes with the latter. These hemoglobin complexes may be degraded on exposure to laser radiation to form free NO that ha a vasodilatory effect. Photolysis of nitrosyl complexes of hemoglobin may be responsible for the therapeutical effect of laser radiation.

[The influence of oxidized fibrinogen on apoptosis of endothelial cells].

Oxidative stress plays an important role in cardio-vascular diseases and atherosclerosis. Fibrinogen (FB), plasma coagulation protein, is a risk factor of atherosclerosis. Importantly, it can be readily oxidized during oxidative stress and in pathological conditions. FB can promote angiogenesis by supporting migration and proliferation of endothelial cells. On the other hand, recent reports demonstrated cytotoxicity of oxidized fibrinogen (oxFB). Endothelial dysfunction plays a critical role in the atherosclerosis development, therefore it is important to understand the effect of oxFB on human endothelial cells (hEC), and the mechanism of the cell death. Here, we studied influence of oxFB on hEC during 24 h incubation in two conditions: (1) at low serum level (0.1%) and in the absence of growth factors ("starvation"); (2) in full medium (5% FBS) with growth factor supplement. Apoptosis was evaluated using analysis of nuclear morphology, phosphatidylserine externalization on hEC surface and caspase-3 activation. In starvation, we observed significant cell death via apoptosis. FB prevented starvation-induced cell death and caspase activation. Caspase activity in the presence of oxFB was 1.5 times higher as compared to FB, yet oxFB demonstrated significant cell protection during stress. Similarly, in optimal cultivation conditions FB decreased the rate of apoptosis by three times, while oxFB supported cell viability to the lesser extent. Thus, FB can protect hEC in stress conditions (in starvation); oxidative modification of FB diminishes its antiapoptotic properties.

[The prospect of pluripotent stem cell-based therapy].

Human embrional stem cells (hESC) are able to maintain pluripotency in culture, to proliferate indefinitely and to differentiate into any somatic cell type. Due to these unique properties, hESC may become an exceptional source of tissues for transplantation and have great potential for the therapy of incurable diseases. Here, we review new developments in the area of embrional stem cells and discuss major challenges--standartization of protocols for cell derivation and cultivation, identification of specific molecular markers, development of new aprouches for directed differentiation etc.--which remain to be settled, prior to safe and successful clinical application of stem cells. We appraise several potential approaches of hESC therapy including derivation of autologous cells via therapeutic cloning (1), generation of immune tolerance to allogenic donor cells via hematopoetic chimerism (2), and development of the banks of hESC lines (3). In addition, we discuss brifly induced pluripotent cells, which are derived via genetic modification of autologous somatic cells and are analogous to ESC. Our analysis demonstrates that uncontrollable differentiation in vivo and teratogenic potential of hESC are critical limitations of their application in clinic. Therefore, the major direction of hESC use is derivation of a specific differentiated progeny, which has lower proliferative potential and immune privilege, yet poses fewer risks. Finally, cell therapy is far more complex and resource-consuming process as compared to drug-based medicine; pluripotent stem cell biology and technology is in need of further investigation and development before these cells can be used in clinics.

Biological activity of hemoprotein nitrosyl complexes.

Chemical and biological functions of hemoprotein nitrosyl complexes as well as their photolysis products are discussed in this review. Chemical properties of nitric oxide are discussed, and major chemical reactions such as interaction with thiols, free radicals, and transition metals are considered. Specific attention is paid to the generation of hemoprotein nitrosyl complexes. The mechanisms of nitric oxide reactions with hemoglobin and cytochrome c and physicochemical properties of their nitrosyl complexes are discussed. A review of photochemical reactions of nitrosyl complexes with various ligands is given. Finally, we observe physiological effects of visible radiation on hemoprotein nitrosyl complexes: smooth muscle relaxation and reactivation of mitochondrial respiration.