Powdery mildew resistance of barley accessions from Dagestan.

Powdery mildew caused by the parasitic fungus Blumeria graminis (DC.) Golovin ex Speer f. sp. hordei Marchal is one of the most common diseases of barley. Growing resistant varieties can signif icantly minimize harmful effects of the pathogen. The specif icity in the interaction between the fungus and its host plant requires a continuous search for new donors of the resistance trait. The powdery mildew resistance of 264 barley accessions from Dagestan and genetic control of the trait in resistant forms were studied under f ield and laboratory conditions. Forty-seven barley lines carrying previously identif ied powdery mildew resistance genes were also examined. During three years, the experimental material was evaluated under severe infection pressure at the Dagestan Experiment Station of VIR (North Caucasus, Derbent). Juvenile resistance against the Northwest (St. Petersburg, Pushkin) pathogen population was evaluated in a climatic chamber. The genetic control of B. graminis resistance in the selected accessions was studied with the application of hybridological and molecular analyses. The level of genetic diversity of Dagestan barley for effective resistance to powdery mildew is very low. Only two accessions, VIR-23787 and VIR-28212, are resistant against B. graminis at both seedling and adult plant stages. The high-level resistance of breeding line VIR-28212 originating from barley landrace VIR-17554 (Ep-80 Abyssinien) from Ethiopia is controlled by the recessive gene mlo11. Accession VIR-17554 is heterogeneous for the studied trait, with the powdery mildew resistant genotypes belonging to two varieties, dupliatrum (an awnless phenotype) and nigrinudum (an awned phenotype). In accession VIR-23787, a recessive resistance gene distinct from the mlo11 allele was identif ied. This accession is supposed to be protected by a new, effective pathogen resistance gene.

[Results of palliative thermochemotherapy for colorectal cancer metastases to the liver].

The purpose of the study is to investigate the effectiveness of thermochemotherapy in patients with colorectal cancer metastases to the liver in palliative mode. Loco-regional hyperthermia in combination with chemotherapy was applied to 33 patients. The comparison group was of 40 patients who underwent only chemotherapy. 5-fluorouracil (5-FU) or capecitabine over standard regimens were used for chemotherapy in both groups. Loco-regional hyperthermia was carried out by capacitive method for 40-90 minutes at a maximum temperature within 41-44 ° C in tumor. No significant differences in dynamics of the overall status, clinical symptoms of the disease and biochemical parameters between the groups were revealed. In the group of patients with thermochemotherapy there were received a complete response in 3 patients (9%), partial response - in 14 patients (44%) while in the group of patients with chemotherapy - in 1 patient (3%) and 11 patients (28%), respectively. The significant difference was found in terms of objective response, i.e. summarized parameters of complete and partial regression (p=0.04). Thermochemotherapy compared with chemotherapy significantly increased survival rates of patients.

[Possibilities of local hyperthermia in treatment for edematous breast cancer patients].

The paper presents an analysis of literature on the results of treatment of edematous breast cancer as well as own data on the use of local hyperthermia for this pathology. Local hy- perthermia in combination with chemotherapy (thermochemo- therapy) was applied in 14 patients with secondary edematous breast cancer. Age of patients ranged from 37 to 72 years (mean 53 years). The study results indicate promising applica- tions of thermochemotherapy in edematous breast cancer: for operable patients as neoadjuvant, for inoperable patients as palliative method of treatment.

[Forensic medical diagnostics of the impact of the ligatures made from different materials in the cases of strangulation asphyxia].

The identification of the instruments of injury resulting from neck compression not infrequently encounters difficulties. We developed an experimental model allowing to evaluate the impact of ligatures made from different materials on non-biological objects. The use of spectral techniques including X-ray fluorescence spectrometry and emission spectral analysis made it possible to detect and identify the chemical elements transferred from the loop material onto the object's surface. Specifically, the transfer of the elements contained in the rubber of the electric cable braid was demonstrated. The presence of these elements together with microparticles of rubber at the surface of the study objects proved to be a diagnostic sign giving evidence of the impact of the rubber sheath of the electric cable leaving the strangulation mark on the object's surface.. The results of the present experiments confirm the possibility of using spectral methods along with other techniques for the identification of a ligature employed as the instrument of strangulation injury.

[The results of combined treatment of patients with locally advanced laryngeal cancer using preoperative chemoradiotherapy].

The combined treatment with preoperative chemoradiotherapy (CRT) was performed in 28 patients with locally advanced laryngeal cancer (T3-4N0-3M0). Radiation therapy (RT) was carried out according to the scheme "1 + 1 Gy" (interval 4-5 hours) 5 times a week till 30-40 Gy. RT was accompanied by simultaneous polychemotherapy: vincristine (1.4 mg/m² per day), cisplatin (20 mg/m²--2-4 days) bleomicetin (10 mg/ m²--5, 6 days), cyclophosphamide (200 mg/m²--7, 8 days). Surgical treatment was carried out in 2.5-3 weeks after CRT. CRT allowed conducting organ-saving surgery on the larynx in 11 (52%) of 21 patients with T3 of primary tumor. All other patients underwent laryngectomy at a different volume. Five-year disease-free survival for the whole group in total (T3-4) was 88%. Relapses of regional metastases (MTS) occurred in 7 of 9 patients including in 4 of 5 patients after lymphadenectomy. The overall survival of patients with T3N02 was 71%, with T4N0-3--20%. Thus we have developed a method of treatment allowing to achieve high local relapse-free survival in patients with locally advanced laryngeal cancer. However organ-saving effectiveness of preoperative CRT and its effect on regional MTS remain low, which requires further research in this direction.

[On the method of forensic medical examination of the injuries inflicted and traces left by the objects having the rubber surface].

A method of forensic medical examination of the injuries inflicted and traces left by the objects having the rubber surface is proposed. The method permits to reveal the facts of contact of injurious rubber things with biological and non-biological objects. Its application extends the possibilities for determining the direction of movement and the weight of a vehicle running over the body of the victim.

Resolution of HLA class I sequence-based typing ambiguities by group-specific sequencing primers.

The increasing demand for allele-level human leukocyte antigen (HLA) typing has led the sequence-based typing (SBT) to become the preferred method. In turn, the steady increase in the number of HLA alleles driven by the adoption of SBT as the ultimate typing method leads to the ever increasing number of cis/trans ambiguities. Over the last few years, additional sequencing with the commercially available group-specific sequencing primers (GSSPs) has replaced sequence-specific primer-polymerase chain reaction and group-specific amplification as the means of resolving cis/trans ambiguities in many laboratories. Here we summarize our 3-year experience in designing and utilizing GSSPs for resolution of HLA class I ambiguities. The panel of GSSPs used in our laboratory includes 14 primers for HLA-A, 18 for HLA-B, and 13 primers for HLA-C. The panel resolves 99.9% of all ambiguities.

[Application of spectral methods for the identification of rubber-sole footwear as a trauma implement].

Kicking to death is estimated to account for nearly 0.5% of all forensic medical autopsies and an appreciable share of murders. Meanwhile, the true number of such cases seems to be much in excess of these statistical data due to the difficulty of ascertaining the nature of injuring tools. This paper reports the results of a comprehensive medico-criminalistic investigation of the traces left by kicking with rubber-sole footwear. It is shown that in certain cases the use of spectral techniques (X-ray fluorescence and emission-spectral analysis) in combination with autopsy data, results of comparative forensic biological studies, and inquest information makes it possible to identify a concrete trauma weapon.

[Attention deficit hyperactivity disorder: the role of parent and teacher questionnaire in the assessment of patient's social-psychological adaptation].

Parents and teachers of 342 school children, aged from 7 to 11 years, were questioned with the <> (SDQ) using non-randomized method. The following comparative analysis of SDQ items measured in parents and teachers of children with ADHD and age-matched controls was carried out. It has been shown that the spectrum of disturbances characteristic of ADHD is not confined only to main symptoms of ADHD. The results of questionnaire of both parents and teachers indicated the marked intensity of emotional disturbances, behavioral problems and difficulties in interactions with peers as well as the underdevelopment of social trends in behavior in children with ADHD compared to the controls. The data obtained confirm the necessity of treatment extension beyond the core symptoms with considering more general parameters of the patients quality of life.

New HLA-C alleles identified in minority populations.

Twenty new human leukocyte antigen C alleles identified in 2200 minority individuals characterized by sequence-based typing.

[Genetic control of the wheat Triticum monococcum L. resistance to powdery mildew].

Using hybrid analysis and test-clone method, 102 accessions of Triticum monococcum L. from the collection of the Vavilov All-Russia Institute of Plant Industry have been studied. This species of wheat has been found to by considerably polymorphic with respect to the resistance to the fungus Erysiphe graminis DC. f. sp. tritici Marchal. causing powdery mildew. The resistance of most accessions to the fungus population and clones is determined by dominant genes. In rare cases, the resistance was determined by recessive genes or one, two, or three oligogenes. A group of einkorn wheat accessions has been found in which the resistance to powdery mildew was determined by the same dominant factor or different but closely linked ones. Recessive resistance genes of T. monococcum differ from the recessive gene pm5 determining the resistance of T. aestivum plants. The genome of T. monococcum contains various genes of resistance to powdery mildew and is a potential source of effective genes to be used when selecting cultivated species of wheat for immunity.

The recombinant HLA-B*5518 allele supports the evidence of conserved haplotype association of rare alleles.

Allelic polymorphism of the major histocompatibility complex arises mostly from gene recombination. Intralocus gene recombination usually involves short fragments of DNA leading most commonly to single-nucleotide substitutions and rarely involves large fragments. Here, we report a new recombinant human leukocyte antigen (HLA)-B*5518 allele that has arisen via recombination of a large fragment of DNA spanning more than 70 nucleotides. During routine HLA typing of potential volunteer donors for the National Marrow Donor Program((R)), a new HLA-B allele was identified in two donors from Guam. The allele, B*5518, appears to be a product of recombination between B*5502 and B*40. Exons 1, 3, and 4 of the new allele belong to B*5502, whereas part of exon 2 belongs to one of B*40 alleles. Introns 1 and 2 appear to belong to B*55, suggesting that the recombination event may have occurred within the homologous parts of exon 2.

Emerging new alleles suggest high diversity of HLA-C locus.

Human leukocyte antigen (HLA)-C has only recently emerged as an important transplantation antigen and as a receptor for natural killer cells. Over the last few years, sequence-based typing (SBT) revealed the true diversity of HLA-C locus; however, the frequency at which new alleles are detected still remains high. During routine SBT of 3500 samples for the National Marrow Donor Program, we have identified 20 new HLA-C alleles reported in this article in 26 individuals. New variants have been characterized by direct sequencing of polymerase chain reaction product obtained by allele-specific amplification of potential new alleles. Most of the new alleles carry coding substitutions of residues located within the antigen-binding groove. The substitutions are predominantly located in the alpha2-helix which is consistent with the unique to HLA-C conservation of alpha1-helix. Seven new alleles, or 35%, have been identified in African Americans, two of them in three and four individuals each, suggesting that these alleles may not be rare. This observation reflects the fact that the minority groups, previously under-represented in the HLA research pools subjected to SBT, now begin to emerge as a main source of new HLA-C alleles. This study further confirms that HLA-C locus is at least as polymorphic as HLA-A and HLA-B.

HLA-Cw*1214 allele arisen via recombination between HLA-Cw*070201 and HLA-Cw*120201.

Allelic polymorphism of the major histocompatibility complex arises mostly from gene conversion. Intralocus gene conversion usually involves limited fragments of DNA, whereas recombination involving large fragments of DNA is considered to be a rare event. During routine sequencing-based typing of donors for the National Marrow Donor Program, a new HLA-C allele was identified in a Caucasian donor. The allele, HLA-Cw*1214, proved to be the product of recombination between HLA-Cw*070201 and HLA-Cw*120201. Exons 1, 2, the 3' end of exon 3 and exon 4 (with one mismatch) belong to HLA-Cw*120201, whereas part of exon 3 belongs to HLA-Cw*070201. Sequencing with primers based in exon 2 and exon 3 showed that intron 2 of the new allele also belonged completely to HLA-Cw*1202. The recombination event apparently occurred within exon 3 with the first point of recombination somewhere between codons 92 and 134 and the second one between codons 157 and 181.

Identification of novel HLA class I alleles using single allele sequencing.

Three new HLA-A and five HLA-B alleles reported in this paper have been characterized by direct sequencing of PCR product obtained by group-specific amplification of potential new alleles. Four new alleles, B*5133, B*5134, B*1574 and B*5807, carry motifs observed in previously identified HLA-B alleles and may have evolved via gene conversion. Four alleles, A*2438, A*3405, A*2437 and B*520104, display polymorphisms at positions previously considered constant. All new alleles were identified either by an unexpected sequence specific oligonucleotide probe hybridization pattern or by sequence-based typing, and later confirmed by single allele sequencing.

Inhibition of cytokine gene expression by sodium salicylate in a macrophage cell line through an NF-kappaB-independent mechanism.

Macrophage-derived cytokines and chemokines are involved at multiple steps of immune and inflammatory responses, and the transcriptional factor NF-kappaB appears to play a pivotal role in their coordinated upregulation. The anti-inflammatory agents salicylates have been proposed to act in part by inhibiting NF-kappaB. We have therefore studied the effects of sodium salicylate on lipopolysaccharide (LPS)-induced kappaB-binding activity and on cytokine and chemokine gene expression in the RAW264.7 murine macrophage cell line and compared them to those of an established NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC). PDTC (100 microM) completely abrogated LPS-induced kappaB-binding activity and also profoundly inhibited the induction of interleukin 1alpha (IL-1alpha), IL-1beta, IL-6, IL-10, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and MCP-1 and, to a lesser extent, leukemia inhibitory factor, RANTES, and IL-1Ra. In contrast, sodium salicylate (15 to 20 mM) had no effect on NF-kappaB activation but, nevertheless, suppressed several LPS-induced cytokine and chemokine genes to a degree similar to that obtained with PDTC. However, compared to PDTC, sodium salicylate caused significantly less inhibition of IL-1Ra and IL-10 gene expression after LPS stimulation. Neither LPS-induced MIP-1alpha, MIP-1beta, nor MIP-2 was significantly affected by PDTC or sodium salicylate, demonstrating that NF-kappaB is dispensable for the transcriptional regulation of these genes by LPS. In summary, these results suggest that both NF-kappaB-dependent and NF-kappaB-independent pathways are necessary for the induction by LPS of a complex cytokine and chemokine response. In the RAW264.7 macrophage cell line, suprapharmacological concentrations of sodium salicylate exert a potent inhibitory effect on LPS-induced cytokine gene induction but appear to accomplish this by interfering with NF-kappaB-independent pathways of activation.

[Inhibition of formation of complement C5-convertase]. / Ingibirovanie obrazovaniia C5-konvertazy komplementa.

Control of the formation of complement C5-convertase is an important factor in maintaining human homeostasis. Some physiologically active endogenous and exogenous substances were shown to be capable of exerting such control. A method was developed for studying the influence of the effectors on the formation of classical-complement-pathway C5-convertase from C3-convertase stabilized by nickel ions. Immunoglobulins G and M, recombinant alpha and gamma interferons, serotonin, epinephrine, histamine, and ethanol were studied as effectors. The binding constants of the effectors with C3b, the active C3 component, and the values of the maximal effect were determined to evaluate the physiological importance of the effectors. The inhibition of C5-convertase generation was observed in all cases except for IgG and IgM (rheumatoid factor), which showed no effect and activating effect, respectively.

[Detection of rheumatoid factor in the blood of patients by latex agglutination]. / Opredelenie revmatoidnogo faktora v syvorotke krovi bol'nykh metodom lateksnoi aggliutinatsii.

A highly effective rapid method is developed for detecting the rheumatoid factor in human serum. Polystyrene suspension-based latex conjugate is synthesized, representing a styrene and methacrylic acid copolymer. The immunospecific reagent is human IgG. The resultant latex conjugate can be used in practical medicine for analysis of the rheumatoid factor.