[Main factors of remodeling of vascular wall].

With a purpose of identifying and ranking factors influencing remodeling of vascular wall in 168 practically healthy subjects (71 men, 97 women) aged 30-60 years we conducted a study of risk factors of cardiovascular disease, and infectious, immunological, inflammatory and metabolic parameters. Using artificial neural network we found that process of remodeling of vascular wall was most significantly impacted by: cytomegalovirus and hlamydia neumoniae infections, decreased number of T-lymphocytes, development of latent vascular microinflammation, and diastolic blood pressure above 80 mm Hg.

[The state of arterial walls in patients with different characeter of atherosclerotic process and latent infection].

Examination of patients differing in manifestation of the atherosclerotic process revealed stable clusters of parameters characterizing the state of arterial walls, presence of infection, immune status, metabolic patterns, and risk factors. Relationship between these variables suggests the dependence of the atherosclerotic process on latent infection, reduced reactivity of the immune system and the action of metabolic factors.

Synthesis of N3-substituted thymidine analogues for measurement of cellular kinase activity.

N3-Substitued thymidine analogues that carry a carboranylalkyl moiety at the N3-position with various spacer lengths have been reported to be good substrates for thymidine kinase (TK1). As part of our continuing effort towards the development of new TK1 substrates for imaging tumor proliferative activity, we have synthesized a series of new N3-substituted analogues of thymidine that carry an aromatic ring with different spacer lengths. The overall yields for 6 and 7 were 13% and 39% in four steps and three steps, respectively, and those for 14, 16 and 18 were in the range of 13%-15% in six steps. The overall yield for 24 was 33% in three steps, and those for 25 and 26 were 64% and 58%, respectively, in one step. Most of these compounds have been tested for TK1 activity by enzymatic assay to identify a good substrate that can be radiolabeled for imaging. The phosphorylation rates of these compounds were 2%-6% compared with that of thymidine. The results from the in vitro enzymatic assays suggest that these N3-substituted thymidine analogues have some potential for imaging TK1 activity if radiolabeled with a suitable isotope.

Molecular imaging of EGFR kinase activity in tumors with 124I-labeled small molecular tracer and positron emission tomography.

Positron emission tomography (PET) with epidermal growth factor receptor (EGFR) kinase-specific radiolabeled tracers could provide the means for noninvasive and repetitive imaging of heterogeneity of EGFR expression and signaling activity in tumors in individual patients before and during therapy with EGFR signaling inhibitors. We developed the synthesis and (124)I-radiolabeling of the (E)-But-2-enedioic acid [4-(3-[(124)I]iodoanilino)-quinazolin-6-yl]-amide-(3-morpholin-4-yl-propyl)-amide (morpholino-[(124)I]-IPQA), which selectively, irreversibly, and covalently binds the adenosine-triphosphate-binding site to the activated (phosphorylated) EGFR kinase, but not to the inactive EGFR kinase. The latter was demonstrated using in silico modeling with crystal structures of the wild type and different gain-of-function mutants of EGFR kinases. Also, this was demonstrated by selective radiolabeling of the EGFR kinase domain with morpholino-[(131)I]-IPQA in A431 human epidermoid carcinoma cells and Western blot autoradiography. In vitro radiotracer accumulation and washout studies demonstrated a rapid accumulation and progressive retention postwashout of morpholino-[(131)I]-IPQA in A431 epidermoid carcinoma and in U87 human glioma cells genetically modified to express the EGFRvIII mutant receptor, but not in the wild-type U87MG glioma cells under serum-starved conditions. Using morpholino-[(124)I]-IPQA, we obtained noninvasive PET images of EGFR activity in A431 subcutaneous tumor xenografts, but not in subcutaneous tumor xenografts grown from K562 human chronic myeloid leukemia cells in immunocompromised rats and mice. Based on these observations, we suggest that PET imaging with morpholino-[(124)I]-IPQA should allow for identification of tumors with high EGFR kinase signaling activity, including brain tumors expressing EGFRvIII mutants and nonsmall-cell lung cancer expressing gain-of-function EGFR kinase mutants. Because of significant hepatobiliary clearance and intestinal reuptake of the morpholino-[(124)I]-IPQA, additional [(124)I]-IPQA derivatives with improved water solubility may be required to optimize the pharmacokinetics of this class of molecular imaging agents.

[Cellular activity in healthy subjects with cardiovascular integral risk factors]. / Pokazateli kletochnoi aktivnosti u prakticheski zdorovykh lits, imeiushchikh integral'nye faktory riska serdechno-sosudistykh zabolevanii.

The study was made of permeability of erythrocytic membrane (PEM), electrokinetic properties of cellular nucleus (ECN) and phagocytic activity of neutrophils (PAN) in whole blood of 130 healthy subjects with integral risk factors of cardiovascular disease. First-level test immunogram was analysed. Hypercholesterolemia and hyperbetalipidemia were associated with high PAN. Obese patients with cardiovascular hereditary load had elevated PAN and PEM. In smokers PEM was elevated but ECN was subnormal, PAN rose in parallel with number of cigarettes smoked for a day. Combination of the risk factors led to low activity of the nucleus and high PAN. It is concluded that healthy subjects with integral risk factors develop changes in functional cell activity.

[Changes in the ligand affinity of benzodiazepine receptors in the presence of 4,5,6,7-tetrahydroisoxazolo(5,4-c)pyridin-3-ol (THIP)]. / Izmenenie affinnosti ligandov benzodiazepinovykh retseptorov v prisutstvii 4,5,6,7-tetragidroizoksazolo-(5,4-c)-piridin-3-ola (TGIP).

4,5,6,7-tetrahydroisoxazolo-(5,4-c-)pyridin-3-ol (THIP) in concentrations of 100-500 microM inhibited 3H-flunitrazepam (3H FNZ) binding to intact membranes at 0 degrees C in 25 mM TRIS-HCl buffer. The inhibition of 3H FNZ binding was due to the decrease in the affinity and Bmax of 3H FNZ binding. The affinity for benzodiazepines (e.g. BZ-receptor agonists) was reduced in the presence of THIP, whereas the affinity for BZ-receptor antagonists was unchanged. The value of THIP-induced shift was dependent on the molarity of TRIS-HCl buffer and the concentration of THIP. The results obtained suggest that THIP-induced shifts have a predictive value in the determination of pharmacological efficacy of BZ-receptor ligands.