[Magnetoencephalography in the presurgical evaluation of patients with drug-resistant epilepsy].

Magnetoencephalography (MEG) in combination with structural MRI (magnetic source imaging, MSI) plays an increasingly important role as one of the tools for presurgical evaluation of medically intractable focal epilepsy. The aim of the study was to compare the MSI and commonly used video EEG monitoring method (vEEG) in their sensitivity to interictal epileptic discharges (IED) in 22 patients with drug resistant epilepsy. Furthermore, the detection and localization results obtained by both methods were verified using the data of electrocorticography (ECoG) and postsurgical outcome in 13 patients who underwent invasive EEG monitoring and surgery. The results showed that MSI was superior to vEEC in terms of sensitivity to IED with difference in sensitivity of 22%. The data also suggested that MSI superiority to vEEG in detecting epileptic discharges might, at least partly, arise from better MEG responsiveness to epileptic events coming from the medial, opercular and basal aspects of cortical lobes. MSI localization estimates were in the same cortical lobe and at the same lobar aspects as the epileptic foci detected by ECoG in all patients. Thus, magnetic source imaging can provide critical localization information that is not available when other noninvasive methods, such as vEEG and MRI, are used.

[Developmental continuity of working memory from infancy to preschool age].

The objective of the paper was to study the developmental continuity of working memory function from infancy to preschool age. At the age of 10 to 11 months 44 participants completed delayed response task (A-not-B) that measures working memory function. Between 5 and 7 years of age the same participants performed three tasks assessing working memory for temporal order in auditory and visual modalities and a control task measuring short-term visuospatial memory. The dependence of temporal-order memory at preschool age on individual level of infant working memory was found for all methods of measurement despite the differences in way of presentation and reproducing of the stimuli order. Results indicate direct continuity in the development of working memory function from infancy to preschool age.

Alterations in stress-associated behaviors and neurochemical markers in adult rats after neonatal short-lasting local inflammatory insult.

Recently, there has been a growing interest in long-term consequences of neonatal pain because modern neonatal intensive care units routinely employ procedures that cause considerable pain and may be followed by local inflammation and hyperalgesia lasting for several hours or even days. To address this question, we developed a rat model of short lasting (<2 days) early local inflammatory insult produced by a single injection of 0.25% carrageenan (CAR) into the plantar surface of a hindpaw. Previously, we demonstrated that rats receiving this treatment within the first week after birth grow into adults with a global reduction in responsiveness to acute pain. Here, we report that these animals also manifest a low anxiety trait associated with reduced emotional responsiveness to stress. This conclusion is based in the following observations: (a) rats in our model display reduced anxiety on an elevated plus-maze; (b) in the forced swim test, these rats exhibit behavioral characteristics associated with stronger ability for stress coping; and (c) these animals have reduced basal and stress-induced plasma levels of such stress-related neuroendocrine markers as corticotropin-releasing factor, vasopressin, and adrenocorticotrophic hormone. In addition, we used DNA microarray and real-time reverse-transcriptase polymerase chain reaction to profile long-term changes in gene expression in the midbrain periaqueductal gray (PAG; a region involved in both stress and pain modulation) in our animal model. Among the affected genes, serotonergic receptors were particularly well represented. Specifically, we detected increase in the expression of 5-HT1A, 5-HT1D, 5-HT2A, 5-HT2C and 5-HT4 receptors. Several of these receptors are known to be involved in the anxiolytic and analgesic activity of the PAG. Finally, to determine whether neonatal inflammatory insult induces elevation in maternal care, which may play a role in generating long-term behavioral alterations seen in our model, we examined maternal behavior for 3 days following CAR injection. Indeed, we observed a substantial increase in maternal attention to the pups at the time of inflammation, but this increase was not without its cost: a period of significant maternal neglect afterward.

[Cloning a sequence, coding an SKT1 family protease inhibitor from potato]. / Klonirovanie posledovatel'nosti, kodiruiushchei ingibitor proteinaz semeistva SKT1 iz kartofelia.

Seven structurally similar clones from potato (Solanum tuberosum L.), cv. Istrinskii genomic DNA were isolated by cloning of the PCR products. It was suggested that five of these clones were the amplified copies of the same gene. Based on comparative and structural analysis of these clones, initial nucleotide structure of the gene was reconstructed. It appeared to be highly homologous (98%) to the already published sequences encoding the proteins belonging to the soybean Kunitz trypsin inhibitor family (SKTI). Comparison of the results with the previously published data on the SKTI-type proteinase inhibitors from potato of cv. Istrinskii suggests that the gene examined encodes both chains of the earlier described PSPI-21-6.3 protein [9].

[Deletion analysis of the structural-functional organization of metalloendopeptidase precursor in B. amyloliquefaciens]. / Izuchenie strukturno-funktsional'noi organizatsii predshestvennika metalloéndopeptidazy B. amyloliquefaciens metodom deletsionnogo analiza.

Functional destination of propeptides and precursors in bacillar secretory proteases remains uncertain. Formerly deletion assay demonstrated folding and secretion of subtilisin E, chymotrypsin-like protease SGPB from S. griseus and B. cereus metalloprotease to depend on full-length propeptide in the precursors. Actually an artificial B. amyloliquefaciens metalloprotease gene with deletion of 51 amino acid residues from N-terminus was constructed with regard to carry out functional mapping of secretory metalloprotease propeptides. B. subtilis wprA gene 5'-terminal region spanning promoter and secretory leader was coupled to provide transcription to the truncated gene and secretion to its product. B. subtilis clones bearing a plasmid with the modified gene synthesised an active mature metalloprotease.

Expression of bacillar glutamyl endopeptidase genes in Bacillus subtilis by a new mobilizable single-replicon vector pLF.

The pLF1311 natural plasmid from Lactobacillus fermentum 1311 was used to construct a single-replicon vector suitable for rapid cloning in a wide range of gram-positive hosts and Escherichia coli. The new vector is capable of conjugative mobilization from E. coli to various hosts by conjugal transfer. The final vector (3.4 kb) showed a high segregational and structural stability and a high copy number. Glutamyl endopeptidase genes from Bacillus licheniformis (gseBL) and B. intermedius (gseBI) were cloned in both pLF9 and pLF14 vectors and introduced to B. subtilis. The yield of enzymes in the pLF-derived producers was 6- to 30-fold more than in the natural producers and reached 100-150 mg/L of mature protease.

Introduction of a mini-gene encoding a five-amino acid peptide confers erythromycin resistance on Bacillus subtilis and provides temporary erythromycin protection in Proteus mirabilis.

A 15-bp mini-gene was introduced into Bacillus subtilis and into stable protoplast-like L-forms of Proteus mirabilis. This mini-gene encoded the peptide MVLFV and modeled a fragment of Escherichia coli 23S rRNA responsible for E. coli erythromycin (Ery) resistance. Expression of the introduced mini-gene conferred permanent Ery resistance on B. subtilis. In L-forms of P. mirabilis, the Ery-protective effect was maintained in the course of several generations. Herewith, the mechanism of Ery resistance mediated by expression of specific short peptides was shown to exist in evolutionary distant bacteria. Three new plasmids were constructed containing the gene under study transcriptionally fused with the genes encoding glutamylendopeptidase of Bacillus licheniformis or delta-endotoxin of Bacillus thuringiensis. The Ery resistance pentapeptide (E-peptide) mini-gene served as an efficient direct transcriptional reporter and allowed to select bacillar glutamylendopeptidase with improved productivity. The mini-genes encoding E-peptides may be applied as selective markers to transform both Gram-positive and Gram-negative bacteria. The small size of the E-peptide mini-genes makes them attractive selective markers for vector construction.

A new PCR-based approach to a fast search of a wide spectrum of cry genes from Bacillus thuringiensis strains.

A pair of highly degenerated primers was adapted to carry out a single-step PCR-detection of any known and probably unknown cry genes of classes cry1, cry4 and cry9 encoding for 130 kDa protein delta-endotoxins in the natural Bacillus thuringiensis (BT) strains. The Southern hybridization of the product has demonstrated that essentially remote cry-genes like cry1Aa and cry9A (cryIG) could be represented in the single amplificate if they are simultaneously present in the genome of the analyzed strain. Four genes were detected by the proposed scheme in the BT ssp. galleriae 11-67. One of them, gene cry1Ga1 was originally found and cloned using the PCR-amplification product obtained from the genomic DNA of this strain as a probe. The new gene was completely identical to one cloned by B. Lambert (unpublished, EMBL accession number Z22510) and essentially related to cryIM (EMBL accession number Y09326), renamed according to the new nomenclature as cry1Ga2.

A novel delta-endotoxin gene cryIM from Bacillus thuringiensis ssp. wuhanensis.

A new cryI-related sequence designated cryIM was cloned using an immunoscreening technique from ssp. wuhanensis of Bacillus thuringiensis (BT), previously reported to produce multiple Cry proteins [Chestukhina et al. (1994) Can. J. Microbiol. 240, 1026-1034]. Analysis of the cryIM nucleotide sequence revealed an ORF, BTII-type promoter-like sequence, peculiar for such genes, a translation initiation element and a putative transcription terminator. Nevertheless, its product was not previously found in the crystals of the host strain [Chestukhina et al. (1994) Can. J. Microbiol. 240, 1026-1034] which shows its weak or absent natural expression. The amino acid sequence of 1151 residues encoded by the continuous reading frame of cryIM is not identical but is essentially similar to the other delta-endotoxins of the CryI class. An IS231-like sequence was found 400 bp downstream of the cryIM stop codon and a fragment of the cryIAb gene was located 3 kb upstream of its initiator codon in the same orientation. Artificial expression of the cloned gene in E. coli under the control of the lacZ promoter allowed us to obtain its hypothetical protein product.

[Venous blood plasma for the serodiagnosis of syphilis]. / Plazma venoznoi krovi dlia serodiagnostiki sifilisa.

Microtest and enzyme immunoassay with venous blood plasma, provided the ratio 0.1 ml of 2 percent EDTA and 2 ml of venous blood is observed, are sufficiently sensitive and specific. These results may be useful in practical laboratories, during prophylactic check-ups, etc.

[The significance of interval characteristics of the quantitative indices of serologic reactions to syphilis in differentiating positive results]. / Znachenie kharakteristik intervalov kolichestvennykh pokazatelei serologicheskikh reaktsii na sifilis pri differentsiatsii polozhitel'nykh reaktsii.

Statistical processing, carried out by the characteristic intervals method, of the results of serologic tests for syphilis in 48 patients with latent syphilis and in 191 patients with false-positive serologic tests has revealed specific and relatively specific intervals of the titers of anti-Treponema antibodies, precipitines and reagines; these intervals make possible a tentative differential diagnosis before specific tests or in cases when these tests cannot be carried out.