Model of Moderate Hyperhomocisteinemia Associated with Mechanical Injury: Dynamics of Morphometric Parameters of Microcirculatory Vessels.

A model of moderate hyperhomocysteinemia associated with mechanical injury of the musculoskeletal system was developed and experimentally substantiated. The adequacy of this model for studies of morphological and functional regularities is verified. This model can be used for the development of a new concept of evaluation of thrombotic complications of mechanical injury.

Monocytes with Oncogenic Mutation JAK2 V617F as a Tool for Studies of the Pathogenic Mechanisms of Myelofibrosis.

We analyzed previously generated stable monocyte-derived cell line carrying mutation JAK2 V617F. Evaluation of the expression of pro- and antifibrotic factors revealed changes in the production of MMPs and their inhibitors, growth factors, galectin-3, and pentraxin 3 in cells carrying mutation JAK2 in comparison with control non-modified cells.

[Results of ultrasound-guided foam sclerotherapy of the great saphenous vein according to the improved technique].

Foam sclerotherapy is an innovative method of treatment for varicose disease, including its truncal forms, making it possible in outpatient conditions to achieve favourable therapeutic and cosmetic outcomes. Analysed herein are the results of foam sclerotherapy of the great saphenous vein (GSV) and its tributaries in a total of 326 patients presenting with varicose disease of lower limbs (395 GSVs) according to the improved methodology (elevation of the extremity to 60°, crural bandage, use of a cooled sclerosant solution). 6-14 days after the first session of sclerotherapy, the control ultrasound examination confirmed occlusion of the GSV in its femoral segment with no reflux in 94.9% of cases (375 GSVs). At terms from 1 year to 5 years, occlusion was diagnosed in 91.1% of cases. Besides alterations in the ultrasound image of the venous superficial bed of the limb and reduction of the trunk of the GSV and its affluents, there was positive dynamics of the clinical symptomatology of the disease.

[Modern tendencies in surgical treatment of patients with obturation jaundice complicated by hepatic insufficiency].

The article is devoted to the problem of efficacy of the hepatic insufficiency (HI) prophylaxis and treatment methods in obturation jaundice. The role of energy-depending processes and the oxygen deficiency in tissues in the HI occurrence and progressing, the possibilities of medicinal correction of such disorders were studied up. The authors follow the concept of expediency of the oxygen-bearing preparations, which improve the erythrocytes function and oxygen delivery to the tissues in combination with miniinvasive decompression of bilioefferent ways, incorporation into the treatment complex of HI of mechanic genesis.

[An oxygen pressure in hepatic tissue of experimental animals in modelling of obstructive jaundice and its treatment].

Efficacy of application of oxygen-transporting preparations in the treatment of obturation jaundice was studied. In experimental conditions while modelling obturation jaundice in animals there were revealed signs of failure of the oxygen delivery to hepatic cells, shown by significant lowering of the oxygen pressure (pO2) in hepatic tissue. To compensate such revealed disorders it is expedient to apply preparations, which owe oxygen-transporting function, such as perftoran or ozonized isotonic solution of sodium chloride. Incorporating perftoran and ozonized isotonic solution of sodium chloride in conventionally accepted complex impedes the tissue hypoxia occurrence, which starts after biliary system decompression, it promotes restoration of hepatic cells functional state in more early terms.

Design and selection of DMP 850 and DMP 851: the next generation of cyclic urea HIV protease inhibitors.

BACKGROUND: Recent clinical trials have demonstrated that HIV protease inhibitors are useful in the treatment of AIDS. It is necessary, however, to use HIV protease inhibitors in combination with other antiviral agents to inhibit the development of resistance. The daunting ability of the virus to rapidly generate resistant mutants suggests that there is an ongoing need for new HIV protease inhibitors with superior pharmacokinetic and efficacy profiles. In our attempts to design and select improved cyclic urea HIV protease inhibitors, we have simultaneously optimized potency, resistance profile, protein binding and oral bioavailability. RESULTS: We have discovered that nonsymmetrical cyclic ureas containing a 3-aminoindazole P2 group are potent inhibitors of HIV protease with excellent oral bioavailability. Furthermore, the 3-aminoindazole group forms four hydrogen bonds with the enzyme and imparts a good resistance profile. The nonsymmetrical 3-aminoindazoles DMP 850 and DMP 851 were selected as our next generation of cyclic urea HIV protease inhibitors because they achieve 8 h trough blood levels in dog, with a 10 mg/kg dose, at or above the protein-binding-adjusted IC90 value for the worst single mutant--that containing the Ile84-->Val mutation. CONCLUSIONS: In selecting our next generation of cyclic urea HIV protease inhibitors, we established a rigorous set of criteria designed to maximize chances for a sustained antiviral effect in HIV-infected individuals. As DMP 850 and DMP 851 provide plasma levels of free drug that are sufficient to inhibit wild-type HIV and several mutant forms of HIV, they could show improved ability to decrease viral load for clinically significant time periods. The ultimate success of DMP 850 and DMP 851 in clinical trials might depend on achieving or exceeding the oral bioavailability seen in dog.

[Estimation of efficacy of the correction methods for hepatic functional disorders in cholestasis of mechanical nature].

Efficacy of the prophylaxis and treatment methods for hepatic insufficiency in obstructive jaundice was studied. The role of an energy-dependent processes and the tissue deficit of oxygen in the hepatic insufficiency occurrence and progress, possibility of these disorders medicinal correction was delineated. The expediency of introduction of preparations, improving the erythrocytes function and the oxygen delivery to tissues in conjunction with miniinvasive biliary decompression, in therapeutic complex for hepatic insufficiency of mechanical nature was noted.

Inhibition of clinically relevant mutant variants of HIV-1 by quinazolinone non-nucleoside reverse transcriptase inhibitors.

A series of 4-alkenyl and 4-alkynyl-3, 4-dihydro-4-(trifluoromethyl)-2-(1H)-quinazolinones were found to be potent non-nucleoside reverse transcriptase inhibitors (NNRTIs) of human immunodeficiency virus type-1 (HIV-1). The 4-alkenyl-3, 4-dihydro-4-(trifluoromethyl)-2-(1H)-quinazolinones DPC 082 and DPC 083 and the 4-alkynyl-3, 4-dihydro-4-(trifluoromethyl)-2-(1H)-quinazolinones DPC 961 and DPC 963 were found to exhibit low nanomolar potency toward wild-type RF virus (IC(90) = 2.0, 2.1, 2.0, and 1.3 nM, respectively) and various single and many multiple amino acid substituted HIV-1 mutant viruses. The increased potency is combined with favorable plasma serum protein binding as demonstrated by improvements in the percent free drug in human plasma when compared to efavirenz: 3.0%, 2.0%, 1.5%, 2. 8%, and 0.2-0.5% for DPC 082, DPC 083, DPC 961, DPC 963, and efavirenz, respectively.

Nonsymmetric P2/P2' cyclic urea HIV protease inhibitors. Structure-activity relationship, bioavailability, and resistance profile of monoindazole-substituted P2 analogues.

Using the structural information gathered from the X-ray structures of various cyclic urea/HIVPR complexes, we designed and synthesized many nonsymmetrical P2/P2'-substituted cyclic urea analogues. Our efforts concentrated on using an indazole as one of the P2 substituents since this group imparted enzyme (Ki) potency as well as translation into excellent antiviral (IC90) potency. The second P2 substituent was used to adjust the physical and chemical properties in order to maximize oral bioavailability. Using this approach several very potent (IC90 11 nM) and orally bioavailable (F% 93-100%) compounds were discovered (21, 22). However, the resistance profiles of these compounds were inadequate, especially against the double (I84V/V82F) and ritonavir-selected mutant viruses. Further modification of the second P2 substituent in order to increase H-bonding interactions with the backbone atoms of residues Asp 29, Asp 30, and Gly 48 led to analogues with much better resistance profiles. However, these larger analogues were incompatible with the apparent molecular weight requirements for good oral bioavailability of the cyclic urea class of HIVPR inhibitors (MW < 610).

Dyschromatosis universalis hereditaria: report of a case.

The case of a 43-old-year woman who had a generalized asymptomatic pigmentary disorder with onset at about age 20 is presented. Tracing back her family history, we found that her father and six of her siblings had also suffered a similar skin pigmentary defect with onset at the same approximate age. In depigmented lesions, three distinct histopathological features were observed: (1) decreased epidermal melanin content and lower density of melanocytes in the upper dermis; and (3) ultrastructural vacuolar degeneration in the focal melanocytes and in the keratinocytes immediately nearby. No deposit of amyloid was observed in the biopsied skin specimens. In hyperpigmented lesions, the histopathological features included: (1) increased melanin content and high density of melanocytes; (2) few melanophages in the papillary dermis of focal areas, but no vacuolar degeneration of the epidermal cells; and (3) an increased number of melanosomes in the basal and suprabasal keratinocytes. Direct immunofluorescence examination revealed no deposit of immunoglobulins in either the hyperpigmented or depigmented lesions. By indirect immunofluorescence examination, the serum of the patient was found to contain antinuclear antibodies (ANA, IgG, class, homogeneous pattern); however, the maximal positive dilution titer of sera against cultured human cells was much higher in melanocytes (1:500 dilution) than in keratinocytes (1:50 dilution) or fibroblasts (1:10 dilution). The pathogenesis of dyschormatosis universalis hereditaria remains unclear; however, hereditary genetic defects may play an important role in alternating regular melanogenesis, which results in a pigmentary anomaly.

[Expression of keratin 16 in normal and lesional skin of solitary and multiple Bowen's disease using monoclonal antibody Ks 8.12 staining].

Bowen's disease is an intraepidermal squamous cell carcinoma usually consisting of a solitary lesion. However, multiple Bowen's lesions are one of the characteristics of arsenicalism in endemic areas where people drink deep well water containing high concentrations of arsenic along the southwest coast of Taiwan. This work seeks to clarify the differences between multiple Bowen's disease in the blackfoot disease endemic area, and solitary Bowen's disease in a non-endemic area by means of Ks 8.12 monoclonal cytokeratin antibody staining. Ks 8.12 may be regarded as a specific antibody for Keratin 16 in the skin and is used as a marker for hyperproliferation. Our results show that Ks 8.12 staining of normal skin in patients with a solitary Bowen's lesion is patchy and always restricted to the basal cell layer. By contrast, the normal skin of patients with multiple Bowen's lesions shows diffuse Ks 8.12 staining of the basal cell layer and various degrees of staining of the suprabasal layers. Similar results were observed in both solitary and multiple Bowen's lesions showing diffuse Ks 8.12 staining of the epidermis. Our results revealed clear differences in keratin expression between the clinically normal skin of patients with solitary Bowen's disease and that of patients with chronic arsenicalism. Finally the clinically normal skin of patients with multipole Bowen's disease showed characteristic changes in the expression of Keratin 16 in the suprabasal layers.

[Viral hepatitis delta in the republic of Belarus]. / Virusnyi gepatit del'ta v Respublike Belarus'.

26,740 blood donors and persons of high risk groups with respect to HBV infection, residing in different regions of Belarus, were examined for the presence of HBsAg in 1983-1997. Of these, 1372 persons (5.1%) were found to have HBsAg, and out of 1081 HBsAg-positive persons anti-HDV antibodies (Ab) were detected in 96 persons (8.9%). In spite of a decrease in acute virus hepatitis B morbidity and in HBsAg carriership, the occurrence of anti-HBV Ab remained stable during the period of 16 years and was equal, on the average, about 4% among asymptomatic HBsAg carriers. Patients having tuberculosis, rheumatoid arthritis, diabetes mellitus, hematological diseases, chronic hepatitides and cirrhosis of the liver were an important reservoir of HBV and HDV infections for regions with the low level of the spread of HBV. A decrease in the detection rate of anti-HDV Ab in patients with cirrhosis of the liver from 47.6% to 15.4% was noted. In 1991-1997 a decrease in the detection rate of anti-HDV Ab in patients with chronic hepatic lesions in comparison with 1983-1990 was observed, and in the age group older than 50 years this decrease was from 33.3% to 8.3%. This difference was particularly pronounces in patients with cirrhosis of the liver: 53.9% and 7.7% respectively.

The increase in intracellular free calcium associated with IgG gamma 2b/gamma 1 Fc receptor-ligand interactions: role in phagocytosis.

The concentration of cytosolic free calcium, [Ca2+]i, was measured in J774 mouse macrophages by use of the fluorescent indicator quin-2. Resting [Ca2+]i was 87 nM. Addition of a number of specific ligands to the immunoglobulin gamma 2b/gamma 1 Fc receptor resulted in a transient increase in [Ca2+]i, the magnitude of which depended on the extent of receptor aggregation. Monovalent ligands gave only a small Ca2+ signal but blocked cell response to subsequent addition of multivalent ligands. Incubation with antibody-coated erythrocytes raised macrophage [Ca2+]i to micromolar levels. [Ca2+]i changes were only partially inhibited by the absence of external Ca2+, suggesting the release of Ca2+ from internal stores in addition to an influx of external Ca2+. These internal stores were not limited to mitochondria. An optimal range of [Ca2+]i was required for phagocytosis. Buffering [Ca2+]i with quin-2 and treating cells with quinine in the absence of external Ca2+ resulted in inhibition of phagocytosis. Increasing [Ca2+]i to micromolar levels with the calcium ionophore A23187 also resulted in similar inhibitory effects. We suggest the involvement of localized cytosolic Ca2+ gradients in generating the signals necessary for phagocytosis.

Study on blackfoot disease: with special reference to evaluating its cutaneous microcirculatory status.

We evaluated the microcirculatory status of blackfoot disease by skin temperature measurement, laser Doppler flowmetry and capillary microscopy. The results of these assessments revealed a good correlation between the disease stage and the microcirculatory status. No effective therapy other than surgical amputation was recommended before. In this study, we treated this endemic disease with prostaglandin E1 (PGE1) infusion therapy. PGE1 was most effective in the erythematous stage and some minor ulcer with an improvement of microcirculatory status. However, PGE1 had no effect in severe ulcerative (ulcer > 0.5 cm) and gangrenous stages. These microcirculatory improvements foreshadowed the improvement of clinical manifestations. The microcirculatory status after PGE1 treatment demonstrated the effectiveness of the therapy.

Lanosterol 14 alpha-methyl demethylase. Isolation and characterization of the third metabolically generated oxidative demethylation intermediate.

Conditions have been identified which permit metabolic formation of the third oxidized intermediate in the lanosterol 14 alpha-methyl demethylase reaction cascade. Metabolism of either the immediate precursor substrate 3 beta-hydroxylanost-8-en-32-al or lanost-8-ene-3 beta,32-diol under mixed function oxidase conditions affords formation of the intermediate. It must be emphasized that the intermediate can only be detected if saponification procedures are omitted during sterol isolation. Comparative chemical and biochemical studies of the isolated metabolite with 3 beta,15 alpha-dihydroxylanost-8-en-32-al reveal that the metabolite is not the 15 alpha-hydroxylanosterol aldehyde, a putative demethylase intermediate. The metabolite is efficiently converted to the demethylated delta 8,14-diene sterol in the absence of molecular oxygen or NADPH, thus supporting its identity as the final oxidized intermediate in the lanosterol 14 alpha-methyl demethylase cascade. 1H NMR analysis shows a proton resonance at 7.86 ppm consistent with a formyloxy proton. Mass spectral and infrared analysis of the metabolite clearly establish oxygen insertion into the immediate precursor substrate, 3 beta-hydroxylanost-8-en-32-al. Collectively, the biochemical and chemical characteristics of the metabolite support a structural assignment for the metabolite as 14 alpha-formyloxy-lanost-8-en-3 beta-ol.

Mechanistic studies of lanosterol 14 alpha-methyl demethylase: substrate requirements for the component reactions catalyzed by a single cytochrome P-450 isozyme.

Lanosterol 14 alpha-methyl demethylation is a cytochrome P-450-dependent process that proceeds through the oxidative sequence of alcohol, aldehyde followed by decarbonylation with formic acid release. Microsomal metabolism studies shown here indicate that only lanostenols and 32-oxy-lanostenols with unsaturation at either the delta 7 or delta 8 position in the sterol can be demethylated. The 14 alpha-methyl group of either lanostan-3 beta-ol or delta 6 lanostenol is not oxidized to the anticipated C-32 alcohol or aldehyde by the enzyme, nor are the corresponding 32-oxy-lanostanols demethylated when incubated with microsomal preparations. Despite the lack of metabolism, the saturated and delta 6 sterol analogues are effective competitive inhibitors of demethylase activity. Utilizing preferred substrates, comparison of the component reactions of the demethylation sequence shows that both the oxidative function and lyase function are sensitive to common inhibitors and that both activities require NADPH. These findings strongly support the premise that a P-450 isozyme does catalyze each phase of the lanosterol 14 alpha-methyl demethylation sequence. Collectively these results demonstrate the double-bond requirement for both components of the demethylation sequence and suggest that the olefinic electrons at delta 7 or delta 8 but not delta 6 may participate directly during demethylation. This participation may involve stabilizing a transition state intermediate or directing activated oxygen insertion as part of the P-450 monoxygenase mechanism.

Novel 2,2-dioxide-4,4-disubstituted-1,3-H-2,1,3-benzothiadiazines as non-nucleoside reverse transcriptase inhibitors.

Benzothiadiazine non-nucleoside reverse transcriptase inhibitors (NNRTIs) of HIV have been synthesized via a novel process to afford active inhibitors, with the most potent compound exhibiting an IC90 = 180 nM in a whole cell assay. The 2,2-dioxide-1H-2,1,3-benzothiadiazine ring system was constructed in one step from 2-amino-5-chlorobenzonitrile.