Diagnostics of Inherited Metabolic Diseases in Newborns with the Hyperammonemia Syndrome at the Onset of Disease (Pilot Study).

The aim of the study was to develop a diagnostic model that allows with a high degree of probability predicting the development of inherited metabolic disease (IMD) in newborns with the hyperammonemia syndrome at the onset of disease and determine the adequate management tactics for such patients. Materials and Methods: The study included 15 female and 5 male infants with the hyperammonemia syndrome. All the patients underwent a full range of clinical laboratory studies in accordance with the standards of treatment of the underlying disease. Blood ammonia measuring (quantitative assessment) was carried out on the basis of the biochemical laboratory of the Children's City Clinical Hospital No.1 (Nizhny Novgorod, Russia) using a portable PocketChem BA blood ammonia meter (Japan) and Ammonia Test Kit II test strips (Japan) in accordance with the manufacturer's protocol. The confirmation of the IMD diagnosis was carried out using the tandem mass spectrometry (TMS) method and molecular genetic testing (genome sequencing).To check the assumptions, all the patients during the follow-up were randomized retrospectively into two groups depending on the TMS results and molecular genetic testing. Group 1 (n=8) included the patients diagnosed with IMD and group 2 (n=12) included the patients with transient hyperammonemia in the neonatal period. Results: The clinical manifestations of the hyperammonemia syndrome in the neonatal period are most frequently realized in the form of a syndrome of CNS depression of varying degrees up to coma (75%), a convulsive syndrome (55%), vomiting (40%) without significant differences in the frequency of occurrence due to the causes of hyperammonemia. In a third of cases, the onset of the hyperammonemia syndrome occurs in the early neonatal period.The ammonia levels in patients with hyperammonemia caused by IMD are statistically significantly higher than those in patients with transient hyperammonemia (p=0.014) which may serve as the first diagnostic sign of IMD in a newborn.A tendency to a more frequent development of anemia in infants with IMD (p=0.084) has been established which might be due to the compensation of metabolic acidosis. The presence of anemia in combination with clinical features and the level of hyperammonemia increases the risk of IMD.In the course of the study, a diagnostic model was developed for predicting the risk of IMD development in a newborn at the onset of hyperammonemia. Through the discriminant analysis, a statistically significant relationship was established between the level of ammonia at the onset, blood glucose and base deficiency levels, blood lactate, hemoglobin, erythrocyte levels, the average volume of erythrocytes and the pH level (p=0.040) with the risk of IMD development. The sensitivity of the model has amounted to 87.5%, the specificity - 83.3%. The diagnostic efficiency of the model is 85.0%. Conclusion: The proposed model can be used at detecting the debut of the hyperammonemia syndrome in newborns in order to predict the probability of IMD and work out the adequate tactics of the management for these patients.

[Main results of research reported by the Center's Department of Innovation in Radiotherapy].

The efficacy of multimodality conservative treatment for prostate and bladder cancer and cervical carcinoma was improved due to integration of such modem modalities as intra-arterial chemotherapy, local hyperthermia and hyperglycemia and combination of local and systemic radiomodifiers. Our methods use criteria of actual survival and are intended to raise it.

Expression genetics and haplotype analysis reveal cis regulation of serine carboxypeptidase I (Cxp1), a candidate gene for malting quality in barley (Hordeum vulgare L.).

Using a cDNA array-based functional genomics approach in barley, several candidate genes for malting quality including serine carboxypeptidase I (Cxp1) were previously identified (Potokina et al. in Mol Breed 14:153, 2004). The gene was mapped as a single nucleotide polymorphism (SNP) marker on chromosome 3H using the Steptoe (feeding grade)xMorex (malting grade) mapping population. Subsequently, the relative level of Cxp1 expression was determined by real-time RT-PCR for each of the 134 progeny lines and mapped as a quantitative trait. Only one quantitative trait locus (QTL) could be identified that significantly influenced the level of the Cxp1 expression. The expressed QTL maps to the same region on chromosome 3H as does the structural gene and corresponds to a QTL for "diastatic power," one among several traits measured to assess malting quality. An analysis of 90 barley cultivars sampled from a worldwide collection revealed six SNPs at the Cxp1 locus, three of which display complete linkage disequilibrium and define two haplotypes. The Cxp1 expression level in a set of barley accessions showing haplotype I was significantly higher than that of accessions displaying haplotype II. The data provide evidence that (1) the expression of Cxp1 is regulated in cis and that (2) the level of diastatic power in the barley seed is influenced by the level of Cxp1 expression.

Molecular orientation in helical and all-trans oligo(ethylene glycol)-terminated assemblies on gold: results of ab initio modeling.

The structural properties of self-assembled monolayers (SAMs) of oligo(ethylene glycol) (OEG)-terminated and amide-containing alkanethiols (HS(CH(2))(15)CONH(CH(2)CH(2)O)(6)H and related molecules with shorter alkyl or OEG portions) on gold are addressed. Optimized geometry of the molecular constituents, characteristic vibration frequencies, and transition dipole moments are obtained using density-functional theory methods with gradient corrections. These data are used to simulate IR reflection-absorption (RA) spectra associated with different OEG conformations. It is shown that the positions and relative intensities of all characteristic peaks in the fingerprint region are accurately reproduced by the model spectra within a narrow range of the tilt and rotation angles of the alkyl plane, which turns out to be nearly the same for the helical and all-trans OEG conformations. In contrast, the tilt of the OEG axis changes considerably under conformational transition from helical to all-trans OEG. By means of ab initio modeling, we also clarify other details of the molecular structure and orientation, including lateral hydrogen bonding, the latter of which is readily possessed by the SAMs in focus. These results are crucial for understanding phase and folding characteristics of OEG SAMs and other complex molecular assemblies. They are also expected to contribute to an improved understanding of the interaction with water, ions, and ultimately biological macromolecules.

Molecular cytogenetic analysis of wheat-barley hybrids using genomic in situ hybridization and barley microsatellite markers.

In the present investigation, genomic in situ hybridization (GISH) and barley microsatellite markers were used to analyse the genome constitution of wheat-barley hybrids from two backcross generations (BC1 and BC2). Two BC1 plants carried 3 and 6 barley chromosomes, respectively, according to GISH data. Additional chromosomal fragments were detected using microsatellites. Five BC2 plants possessed complete barley chromosomes or chromosome segments and six BC2 plants did not preserve barley genetic material. Molecular markers revealed segments of the barley genome with the size of one marker only, which probably resulted from recombination between wheat and barley chromosomes. The screening of backcrossed populations from intergeneric hybrids could be effectively conducted using both genomic in situ hybridization and molecular microsatellite markers. GISH images presented a general overview of the genome constitution of the hybrid plants, while microsatellite analysis revealed the genetic identity of the alien chromosomes and chromosomal segments introgressed. These methods were complementary and provided comprehensive information about the genomic constitution of the plants produced.

[Sociomedical examination and rehabilitation of patients with tuberculosis]. / Mediko-sotsial'naia ékspertiza i reabilitatsiia bol'nykh tuberkulezom.

The paper provides primary and secondary disability rates during 5 years (1993-1997), derived by the tuberculosis agency of a Moscow sociomedical examination commission. Examination was made mainly in patients with pulmonary and extrapulmonary tuberculosis, chronic nonspecific lung diseases, silicosis, cancer, and concomitant diseases of the heart, liver, kidney, etc. There was a rise in primary and secondary disability rates due to the increases of Groups 2 and 3. More than 50% of the patients do not work as unemployment is on the rise in Russia. Group 3 patients were the smallest proportion and temporarily disabled patients were also few.

Evolutionary conservation of kinetochore protein sequences in plants.

The evolutionary conservation of structural/functional kinetochore proteins has been studied on isolated nuclei and pro-/metaphase chromosomes of mono- and dicot plants. The cross-reactivities of antibodies against human CENPC, CENPE and CENPF, and against maize CENPCa with the centromeric regions of mitotic chromosomes of Vicia faba and/or Hordeum vulgare are shown. Putative homologs of the kinetochore protein SKP1 (suppressor of kinetochore protein 1p of yeast) were found in both species and of CBF5p (centromere binding factor 5 of yeast) in barley. Antibodies against synthetic peptides derived from partial sequences encoding these proteins were produced and recognized the centromeric regions on mitotic chromosomes as detected by indirect immunofluorescence.

Calreticulin expression in plant cells: developmental regulation, tissue specificity and intracellular distribution.

The tissue-specific expression pattern and the intracellular distribution of the Ca(2+)-binding protein calreticulin at the mRNA and protein levels have been studied during somatic and zygotic embryogenesis of Nicotiana plumbaginifolia Viv. A full-length cDNA sequence encoding calreticulin was isolated from a lembda Zap cDNA library from early developmental stages of somatic embryogenesis. The deduced amino acid sequence of the calreticulin from N. plumbaginifolia shows high homology to the corresponding proteins of tobacco (98.2% identity), maize (80%) and barley (76.5%), and more than 55% homology to animal calreticulins, and the sequence motifs with established functions found in calreticulins of other species were quite conserved. Northern experiments revealed a developmental regulation of the calreticulin transcript with a maximum during the early stages of somatic embryogenesis and an auxin dependence during in-vitro cell culture. alpha-Naphthaleneacetic acid stimulated calreticulin expression whereas 2,4-dichlorophenoxyacetic acid reduced it. Immunohistological analysis of calreticulin distribution in the ovaries during zygotic embryogenesis showed that calreticulin biosynthesis started tissue specifically, with a high abundance in the endothelium of the integument in the ovules, followed by calreticulin accumulation in the embryo proper and in the associated endosperm at the late globular stage of embryogenesis. Using immunogold labeling, calreticulin was intracellularly localized with a high abundance to the Golgi compartment and to patches on the surface of dividing protoplasts. Smaller amounts were found in the endoplasmic reticulum and plasma membranes. The functional role of calreticulin in posttranslational processing and translocation processes, apart from its postulated function in cellular Ca2+ homeostasis, is discussed.

[Structural-functional changes in the thyroid gland after chronic exposure to the dioxin-containing herbicide 2,4-D]. / Strukturno-funktsional'nye izmeneniia v shchitovidnoi zheleze pri khronicheskom vozdeistvii dioksinsoderzhashchego gerbitsida 2,4-D.

White rats were given by lavage 40% solution of the herbicide 2,4-D (as an amine salt) in doses 1.0 and 0.5 mg/kg during 3 months. Destructive changes in follicles, lymphohistiocytic infiltrates, areas with morphological signs of activation (predominance of small follicles), high alkaline phosphatase and glycoproteins activity in thyrocytes were revealed.

[Morphofunctional characteristics of the testis of albino rats exposed to dioxin-containing herbicide 2,4-D]. / Morfofunktsional'naia kharakteristika semennikov belykh krys pri vozdeistvii dioksinsoderzhashchego gerbitsida 2,4-D.

Endocrine system of the gonads of white rats after subacute poisoning with 2,4-D was studied. Signs of increased steroidogenesis in Leydig cells, a decrease of plasma testosterone concentration and decrease of the gonad sensitivity to chorionic gonadotropin were observed.

A Ty3/gypsy retrotransposon-like sequence localizes to the centromeric regions of cereal chromosomes.

A 745 bp sequence (pSau3A9) located at the centromeres of several cereal species was isolated from a sorghum BAC library by Jiang et al. (1996, Proc. Natl Acad. Sci. USA, 93, 14210-14213). We have amplified a partially homologous 809 bp sequence from barely genomic DNA by PCR and localized it to the centromeres of barley, wheat and rye chromosomes by fluorescent in situ hybridization (FISH). Sequence analysis showed this barley homolog of pSau3A9 to have high similarity to the integrase region of the polyprotein gene of Ty3/gypsy group retrotransposons. Using this integrase sequence as a probe, several clones were isolated from a lambda library constructed of genomic barley DNA. One of the lambda clones contained coding regions for all five catalytic sites characteristic of the retrotransposon polyprotein. Two direct repeats flanking the polyprotein gene are homologous to the cereal centromeric sequence described by Aragón-Alcaide et al. (1996, Chromosoma, 105, 261-268) and may represent all or part of the long-terminal repeats (LTRs). Different plasmid subclones containing various regions of the lambda clone were used in FISH to show that the entire polyprotein gene and upstream flanking sequences, including the presumed LTR, are present at barley centromeres. The preferential (or exclusive) localization of an apparently complete retroelement within the centromeric regions of several cereal species raises interesting questions about its role in karyotype evolution and centromere function.

[Comparative evaluation of fibrogenesis and toxicity of asbestos-containing heat-proof materials]. / Sravnitelnaia otsenka fibrogennosti i tiksichnosti asbestsoderzhashchikh teploizoliatsionnykh materialov.

The article deals with the results of studies analyzing the fibrogenic activity and toxicity of asbestos-containing heat insulators exemplified by asbestos board, vulcanite and asbestos vermiculite. Physical and chemical parameters of the studied dust are presented. All studied dust types have the distinct fibrogenic activity, the highest one was seen in asbestos board. The studied dust types could be classified according to the acute toxicity: asbestos board (DL50-3.2 +/- 0.09 g/kg), chrysotile asbestos (DL50-2.0 +/- 0.08 g/kg), vulcanite (DL50-1.9 +/- 0.08 g/kg) and asbestos vermiculite (DL50-0.7 +/- 0.04 g/kg). The article touches upon the probable acting mechanism of acute toxicity of asbestos and asbestos-containing dust.

[Comparative evaluation of dust oncogenicity in asbestos-containing and non-asbestos details]. / Sravnitelnaia otsenka onkogennosti pylei asbestoformovannykh detalei na bezasbestovoi i asbestovoi osnove.

Experiments proved higher carcinogenicity of chrisotyle-asbestos than of dust appearing in mechanical processing of asbestos-containing details and in wearing articulated shoe brakes. Substitution of asbestos for basalt fibers leaves oncologic risk intact among workers engaged into mechanical processing of asbestos-containing brakes. Avoiding asbestos in mixtures for brakes could lower risk. Results support conclusions obtained by cohort studies in Yaroslavl.

[The effect of the time of methotrexate administration over 24 hours on the antitumor activity of the preparation, its concentration in the blood and in the tumor in Pliss' lymphosarcoma in rats]. / Vliianie vremeni vvedeniia metotreksata v techenie sutok na protivoopukholevuiu aktivnost' preparata, ego kontsentratsiiu v krovi i opukholi pri limfosarkome Plissa u krys.

A mismatch between a circadian rhythm of blood-methotrexate level and that of the drug concentration in rat transplanted lymphosarcoma tissue was established. The drug proved 3 times more effective when given at the time of the day when its concentration in tumor tissue was likely to be the highest.

[Toxicological characteristics and standardization of various poly-metallic oxides]. / Toksikologicheskaia kharakteristika i reglamentatsiia riada polimetallicheskikh oksidov.

Acute toxicity, functional cumulative and local effects of the four ++poly-metallic oxides YBa2Cu3O7, Bi4Sr3O16, Tl2Ca2Cu3O10, Lao,7Sro,3CoO3, were studied in laboratory animals. Parallel experiments on assessment toxicity of the regular oxides of the studied metals in mice were carried out. The major dissolution of metals was shown to occur in acid medium. Thallium-barium-calcium cuprite turned out to be the most toxic and cumulative; it induced the most severe morphologic changes in the liver and kidneys. The least severe disorders caused by this substance were in the rat gastric mucosa and in rabbit eye after a conjunctival injection. Yttrium-barium cuprite induced the most severe changes at the site of injection, lanthanum-strontium cobaltite ranks second here. All copper-containing polymetallic oxides show resorptive toxicity when applied to the skin. Different types of combined toxicity are possible, that is why MACs for the regular oxides can not be used for setting the normal concentrations of ++poly-metallic oxides in air. Estimated toxicometric parameters helped to score the concentrations, which may serve as tentative safe levels of the studies substances.

[Investigation of food poisoning of staphylococcal etiology using a comparison of the extracellular proteins of the isolated causative agents]. / Rassledovanie pishchevykh toksikoinfektsii stafilokokkovoi étiologii s ispol'zovaniem sopostavleniia spektrov vnekletochnykh belkov vydelennykh vozbuditelei.

Materials on the study of four outbreaks of alimentary toxinfections of staphylococcal etiology, carried out by the method of the identification of staphylococci by the spectra of their extracellular proteins, are presented. As the result of this study, the sources and transfer routes of infections have been established. The method of the identification of staphylococci by the exoprotein spectra has made it possible to find out the epidemiological chain with greater accuracy. Such identification may serve as an auxiliary method in epidemiological surveillance.