Blood oxygen transport and pro-oxidant-antioxidant state during liver reperfusion

Indices of blood oxygen transport (hemoglobin-oxygen affinity, pCO2, pH, pO2, etc.) and prooxidant-antioxidant state (Schiff bases, conjugated dienes, catalase, retinal, alpha-tocopherol) were measured in rabbit blood and the liver during postischemic reperfusion. Hepatic ischemia was induced for 30 min by ligation of a hepatica propria, and reperfusion lasted for 120 min. Hepatic ischemia worsened blood oxygen transport. Restoration of arterial blood flow did not result in improvement of oxygen delivery. Moreover, marked metabolic acidosis was observed throughout 2 hr of reperfusion. Ischemia induced a shift of oxyhemoglobin dissociation curve to the right. This shift persisted after restoration of hepatic arterial blood flow facilitating increased oxygen transport to tissues. Changes in blood oxygen transport during hepatic ischemia/reperfusion were accompanied with high activity of free radical processes. During reperfusion, the largest increase in content of lipid peroxidation products and the greatest fall of some antioxidant levels except catalase were observed indicating impairment of liver prooxidant-antioxidant balance. The results showed that activation of lipid peroxidation and a decrease in some antioxidant levels during hepatic reperfusion were associated with lowering of hemoglobin-oxygen affinity and suggest participation of the latter in impairment of prooxidant-antioxidant balance.     

Computer modelling of the organs of the upper abdominal cavity

Method of creation computer models of upper floor of abdomen organs: liver, gallbladder, duodenum, pancreas, stomach, blood vessels with using computer system "DUCT" was described. Details of modelled structures of the cylindric and complicated forms were noted.     

The UTX gene escapes X inactivation in mice and humans

We recently have identified a ubiquitously transcribed mouse Y chromosome gene, Uty , which encodes a tetratricopeptide repeat (TPR) protein. A peptide derived from the UTY protein confers H-Y antigenicity on male cells. Here we report the characterization of a widely transcribed X-linked homologue of Uty , called Utx , which maps to the proximal region of the mouse X chromosome and which detects a human X-linked homologue at Xp11.2. Given that Uty is ubiquitously transcribed, we assayed for Utx expression from the inactive X chromosome (Xi) in mice and found that Utx escapes X chromosome inactivation. Only Smcx and the pseudoautosomal Sts gene on the mouse X chromosome have been reported previously to escape inactivation. The human UTX gene was also found to be expressed from Xi. We discuss the significance of these data for our understanding of dosage compensation of X-Y homologous genes in humans and mice.      

High throughput parallel analysis of hundreds of patient samples for more than 100 mutations in multiple disease genes

As more mutations are identified in genes of known sequence, there is a crucial need in the areas of medical genetics and genome analysis for rapid, accurate and cost-effective methods of mutation detection. We have developed a multiplex allele-specific diagnostic assay (MASDA) for analysis of large numbers of samples (> 500) simultaneously for a large number of known mutations (> 100) in a single assay. MASDA utilizes oligonucleotide hybridization to interrogate DNA sequences. Multiplex DNA samples are immobilized on a solid support and a single hybridization is performed with a pool of allele-specific oligonucleotide (ASO) probes. Any probes complementary to specific mutations present in a given sample are in effect affinity purified from the pool by the target DNA. Sequence-specific band patterns (fingerprints), generated by chemical or enzymatic sequencing of the bound ASO(s), easily identify the specific mutation(s). Using this design, in a single diagnostic assay, we tested samples for 66 cystic fibrosis (CF) mutations, 14 beta-thalassemia mutations, two sickle cell anemia (SCA) mutations, three Tay-Sachs mutations, eight Gaucher mutations, four mutations in Canavan disease, four mutations in Fanconi anemia, and five mutations in BRCA1. Each mutation was correctly identified. Finally, in a blinded study of 106 of these mutations in > 500 patients, all mutations were properly identified. There were no false positives or false negatives. The MASDA assay is capable of detecting point mutations as well as small insertion or deletion mutations. This technology is amenable to automation and is suitable for immediate utilization for high-throughput genetic diagnostics in clinical and research laboratories.      

Mast cells in photolesion of the skin and basal cell cancer associated with it

Morphofunctional features of skin mast cells located in the areas subjected to chronic UV-radiation and in the associated basal cell carcinoma with photoinjure have been studied. Various immunohistochemical methods (chromogranin A, CDla, HLA-DR, CD35, Ki67, P53, Bcl-2, Mcl-1, involucrine) were used. It is found that chronic UV-damage leads to mast cell hyperplasia as well as activation of their synthetic, absorption and secretory functions. It is suggested that mast cell hyperplasia and increase of mast cells neuroendocrine activity provide a risk of basal cell carcinoma development.     

Achievements in morphological diagnosis of prostatic cancer: alpha-methylacyl-coenzyme-A-racemase--a new marker of malignant cell transformation

Gene P504S is considered as the most specific for prostatic carcinoma and its protein (alpha-methylacyl coenzyme A racemaze (AMACR/P504S) is higly sensitive and specific marker not only for adenocarcinoma cells but also for preceding changes - prostatic intraepithelial neoplasm (PIN). AMACR/P504S seems to be the first marker of malignant transformation and tumor progression. Use of immunohistochemical method for revealing this marker together with methods of basal prostatic cells observation (cytokeratin of a high molecular weight, cytokeratin 5/6, p63) improves morphological diagnosis of prostatic carcinoma, particularly on the material of needle biopsies. This combination allows one to identify neoplastic nature of some difficult lesions.     

Morphological features of juvenile colon polyps in children

Histological, histochemical and immunohistochemical studies of 50 solitary juvenile polyps (JP) and 50 JP from children with juvenile polyposis syndrome (JPS) were performed. Observations of the focal complex glandular structures with high mitotic rate were more frequent in JP from patients with JPS (n = 29, 58%) than in solitary JP (n = 17, 34%) (p < 0.03). The immunohistochemical study demonstrated p53 overexpression in individual cells and more than 50% of Ki-67-positive cells in 5 (10%) solitary JP and in 17(34%) JP from patients with JPS (p < 0.007). The finding of microglandular pattern is more typical for JP from patients with JPS. Pathological data, expression of p53 and Ki-67 by immunohistochemistry could help to pick out the group of JP with dysplastic changes.     

Nucleotide sequence of gag and env genes of human immunodeficiency virus type 1, found in Russia: detection of new recombinant variants

Phylogenetic analysis of nucleotide sequences of gag and env genes of type 1 human immunodeficiency virus (HIV-1) variants isolated from individuals infected through sexual intercourse or nosocomially (by injections with nonsterile syringes) showed that 5 of 27 (18.5%) isolated strains were recombinants. Two viruses found in the Russian Far East had gagAenvE genotype, three other recombinants had envG genotype; gag gene of one isolate belonged to subtype A and gag genes of two others belonged to subtype D. Detection of new recombinant variants in addition to the A/B recombinant described previously shows that these viruses can contribute to the HIV-1 genetic variability in Russia.     

Stimulation of β-adrenoceptors inhibits store-operated channel currents via a cAMP-dependent protein kinase mechanism in rabbit portal vein myocytes

Previously we have described the properties of store-operated channel currents (SOCs) in freshly dispersed rabbit portal vein smooth muscle cells. In addition to Ca²⁺ store depletion these SOCs could also be activated by α-adrenoceptor stimulation and diacylglycerol (DAG) via a protein kinase C (PKC)-dependent mechanism. In the present study we have investigated the effect of β-adrenoceptor stimulation on SOCs in rabbit portal vein myocytes. With whole-cell recording the selective β-adrenoceptor agonist isoprenaline reduced the current evoked by cyclopiazonic acid (CPA, sarcoplasmic/endoplasmic reticulum ATPase inhibitor) by over 85%. With cell-attached patch recording, bath application of isoprenaline produced a pronounced inhibition of SOC activity evoked by either CPA or the acetoxymethyl ester form of BAPTA (BAPTA-AM). SOC activity evoked by CPA, the DAG analogue, 1-oleoyl-acetyl- sn -glycerol (OAG) or the phorbol ester, phorbol-12,13-dibutyrate (PDBu) was also markedly inhibited by the adenylate cyclase activator, forskolin, and the cell-permeable non-hydrolysable analogue of cyclic adenosine monophosphate (cAMP), 8-Br-cAMP. With inside-out patches, bath application of PDBu evoked channel currents with similar properties to SOCs which were inhibited by over 90% by a catalytic subunit of protein kinase A (PKA) and by 8-Br-cAMP. Moreover bath application of PKA inhibitors, H-89, KT5720 and an inhibitory peptide to quiescent cell-attached or inside-out patches, activated channel currents with similar properties to SOCs. These data suggest that in rabbit portal vein myocytes, stimulation of β-adrenoceptors inhibits SOC activity via a cAMP-dependent protein kinase signal transduction cascade. In addition it is concluded that constitutive PKA activity has a profound inhibitory effect on SOC activity in this vascular preparation.