[Radiological analysis of size of bone elements of the temporomandibular joint in children and adolescents with class II malocclusion]. / Analiz razmerov kostnykh elementov visochno-nizhnechelyustnogo sustava u detei i podrostkov s distal'noi okklyuziei.

BACKGROUND: The aim of the study was to assess the ratio of bone elements of the temporomandibular joint (TMJ) in children and adolescents with distal occlusion according to cone-beam computed tomography (CBCT). MATERIAL AND METHODS: CBCT of the TMJ was analyzed in 121 patients aged 12-18 years, which were divided into two age groups: group I - children aged 12-15 years (62 cases in total, of which 47 patients with distal occlusion and 15 patients with physiological occlusion) and II group - adolescents aged 15-18 years (59 cases, of which 48 patients with distal occlusion and 11 patients with physiological occlusion). CBCT of the temporomandibular joint was performed by the standard method in the position of habitual occlusion and maximally open mouth. The size of the TMJ joint space in the anterior, upper and posterior sections was determined by the method of Rabukhina N.A., the analysis of the ratio of the bone elements of the TMJ was carried out using the Craniometry program. RESULTS: The size of the joint space and the angle ß of the temporomandibular joint in the anterior section on the right and left in children aged 12-15 with distal occlusion is increased compared to children with physiological occlusion in the position of habitual occlusion. In adolescents 15-18 years old with distal occlusion, the size of the TMJ joint space is increased in the anterior section on the right and left and reduced in the posterior section on the right, angle α is reduced on the right and left, angle ß is reduced on the right, angle γ is increased on the right compared to adolescents with physiological occlusion. The revealed deviations indicate the presence of violations in the ratio of the bone elements of the TMJ, which are aggravated with age. CONCLUSION: Analysis of the results obtained during CBCT in children and adolescents allows us to correct the plan of orthodontic treatment and predict its results.

Draft Genome Sequence of Lactobacillus fermentum NB-22.

We announce here a draft genome sequence of Lactobacillus fermentum NB-22, a strain isolated from human vaginal microbiota. The assembled sequence consists of 190 contigs, joined into 137 scaffolds, and the total size is 2.01 Mb.

Development of Primers and Probes for Genus and Species Specific Detection of 'Candidatus Liberibacter Species' by Real-Time PCR.

Huanglongbing (HLB), also known as citrus greening, is currently the most devastating disease impacting citrus production. The disease is associated with three different 'Candidatus Liberibacter species', 'Ca. Liberibacter asiaticus', 'Ca. Liberibacter americanus', and 'Ca. Liberibacter africanus', which induce similar and overlapping symptoms. When HLB-symptomatic trees are tested, one of the Candidatus Liberibacters is normally detected by conventional or real-time PCR (qPCR). The most widely used assays use primers and probes based on the 16S ribosomal RNA (rRNA) gene. The 16S rRNA-based assays to detect the three species are species-specific and must be performed sequentially. We describe a single assay that detected all species of 'Ca. Liberibacter' at the genus level, providing increased convenience. Recent molecular analyses of 'Ca. Liberibacter species' and other bacteria suggest that the rpoB gene (encoding the ß-subunit of RNA polymerase) provides an alternative target for bacterial identification. We report here the design of a single pair of degenerate primers and a hybridization probe corresponding to the rpoB region and their application for the detection of all three citrus 'Ca. Liberibacter species', enabling detection of 'Ca. Liberibacter' at the genus level. In addition, species-specific primers and probes based on the rplJ/rplK genes were designed and used for detection at the species level in a multiplexed format. Both the genus- and species-specific assays were validated in both SYBR Green I and TaqMan formats, and with both plant and insect extracts that contained the pathogen. These one-step qPCR diagnostic methods are useful for the detection of all species of Liberibacter infecting citrus. In addition, the degenerate genus-specific primers and probe successfully detected 'Ca. Liberibacter solanacearum', a psyllid-transmitted pathogen associated with disease in tomato, carrot, and potato.

Murraya paniculata and Related Species as Potential Hosts and Inoculum Reservoirs of 'Candidatus Liberibacter asiaticus', Causal Agent of Huanglongbing.

Huanglongbing (HLB), considered to be the most serious insect-vectored bacterial disease of citrus, is transmitted in nature by the Asian citrus psyllid Diaphorina citri and the African citrus psyllid Trioza erytreae. D. citri was discovered in southern Florida in 1998 and the HLB disease in 2005. Both have become established throughout citrus-producing areas of Florida. Murraya species are widely grown in southern Florida as ornamental hedges and are readily colonized by D. citri vectors. Colonies of D. citri, isolates of 'Candidatus Liberibacter asiaticus' from Taiwan and Florida, and the Murraya species were established in the BSL-3 biosecurity facility at Fort Detrick. In controlled inoculation experiments, D. citri transmitted 'Ca. L. asiaticus' into M. paniculata (34/36 plants) and M. exotica (22/23 plants), but not into Bergera (Murraya) koenigii. Disease symptoms rarely developed in Murraya plants; however, positive infections were determined by conventional and real-time polymerase chain reaction (PCR). Back-inoculations of 'Ca. L. asiaticus' from M. paniculata to Madam Vinous sweet orange resulted in disease development in 25% of the inoculated plants. Considerable variability was observed in infection rates, titer, and persistence of 'Ca. L. asiaticus' in infected Murraya.

[Use of microstream capnography and alveolar recruitment during off-pump coronary artery bypass grafting].

The aim of the study was to investigate changes in EtCO2 and its correlation with PaCO2, and cardiac function during off-pump coronary artery bypass grafting (OPCAB) and to evaluate whether the recruitment maneuver was effective in improving gas exchange after OPCAB. Twenty adult patients scheduled for elective OPCAB were enrolled in a prospective randomized study. Anesthesia was maintained with midazolam, propofol, and fentanyl. After OPCAB the patients were randomized to a control group receiving conventional ventilation (n=10) or to a RM group (n=10) having ventilation and RM. RM was performed at min 15 after transfer to an ICU, by increasing airway pressure to 40 cm H2O for 40 sec subsequently adjusting PEEP to a level of 2 cm H2O above the lower inflection point of the pressure-volume curve. The measurements included hemodynamics, microstream capnography, respiratory parameters, and blood gasses. The baseline EtCO2 correlated with PaCO2 and cardiac index in both group (r = 0.7 and 0.81, respectively; p < 0.05). In the control group, OPCAB was followed by a rise in PaCO2 and worsening of arterial oxygenation (p < 0.05). After recruitment, EtCO, increased transiently whereas PaO2/FiO2 return to the baseline level. There was a moderate correlation between EtCO2 and PaCO2 before and after RM (r = 0.7 and 0.8, respectively; p < 0.05). The Bland-Altman analysis has shown that the difference between PaCO2 an EtCO2 was 1.9 +/- 11.4 mm Hg (M +/- 2SD). Thus, during OPCAB, EtCO2 measured by microstream capnography cor related well with PaCO2 and cardiac function. The use of RM after OPCAB increases CO2 elimination and improve arterial oxygenation.

Cultivation of 'Candidatus Liberibacter asiaticus', 'Ca. L. africanus', and 'Ca. L. americanus' associated with huanglongbing.

A new medium designated Liber A has been designed and used to successfully cultivate all three 'Candidatus Liberibacter spp.,' the suspect causative agents of huanglongbing (HLB) in citrus. The medium containing citrus vein extract and a growth factor sustained growth of 'Ca. Liberibacter spp.' for four or five single-colony transfers before viability declined. Colonies, positive for 'Ca. L. asiaticus' by a 16s-based rDNA real-time polymerase chain reaction (RT-PCR) assay and sequencing, were irregular-shaped, convex, and 0.1 to 0.3 mm after 3 to 4 days. Suspect 'Ca. L. asiaticus' and 'Ca. L. americanus' cells were observed in infected tissue and on agar culture by scanning electron microscopy. The cells were ovoid to rod shaped, 0.3 to 0.4 by 0.5 to 2.0 microm, often with fimbriae-like appendages. Two strains of 'Ca. L. asiaticus' and one of 'Ca. L. americanus' grown on Liber A medium were pathogenic on citrus and could be isolated from noninoculated tissues of inoculated trees and seedlings 9 and 2 months later, respectively. The identity was confirmed by RT-PCR and 16s rDNA sequencing. This is the first report of the cultivation and pathogenicity of 'Ca. L. asiaticus' and 'Ca. L. americanus' associated with symptoms of HLB.

First Report of Crown Gall Caused by an Agrobacterium sp. on Diffuse Knapweed (Centaurea diffusa).

Diffuse knapweed (DK) plants were discovered in Mosier, Wasco County, OR (45.6842°N, 121.4021°W) with crown gall-like symptoms near the soil line. Specimens were collected on 27 July 2004 and sent to the USDA-ARS at Ft. Detrick, MD for identification of disease and pathogen. Pure culture of a bacterium was obtained on potato dextrose agar, and hyperplasia and hypertrophy developed on carrot disks and tomato stems after wound inoculation with a needle contaminated by the agar culture. The same bacterium was reisolated from the galls on DK, thus fulfilling Koch's postulates. Pathogenicity tests involving needle inoculations of stems and petioles resulted in gall formation on Acroptilon repens, Carthamus tinctorius, Centaurea solstitialis, C. maculosa, C. cyanus, Crupina vulgaris, Helianthus annuus, and Rubus armeniacus. In biochemical tests typically used for identification of Agrobacterium species (3), the DK strain grew on D1M agar but not on 2% NaCl medium, produced acid from erythritol but not from melezitose, converted malonic acid to base, and turned litmus milk alkaline. These results are characteristic of Agrobacterium rhizogenes (= Biovar 2), except for the litmus milk reaction. Using 16S rRNA cluster analysis by unweighted pair group method with arithmetic mean (UPGMA, 500 replicates) and basic local alignment search tool (BLAST), the DK strain clustered most closely with A. rubi (GenBank Accession Nos. D12787 and AM181759). The DK strain differed from A. larrymoorei (GenBank Accession No. Z30542), A. tumefaciens (GenBank Accession No. AJ389896), A. rhizogenes (GenBank Accession No. AB247607), and A. vitis (GenBank Accession No. AB247599) on the basis of 16S rRNA sequence cluster analysis. The DK strain differed from A. rubi on the basis of differential reactions with erythritol, litmus milk, and 2% NaCl medium (2,4); and the 16S rRNA sequence of the DK strain differed from that of A. rubi by 11 bp (99.2% similarity). Comparisons also were made between the DK strain and two strains (83A and 135A) of A. tumefaciens (= Biovar 1), described from New Mexico on A. repens (1), a plant species in the same tribe and subtribe of the Asteraceae as DK. Host range reported for the two A. repens strains after artificial greenhouse inoculations was similar to that of the DK strain and it included diffuse knapweed (1). However, 16S sequencing, which confirmed identification of both A. repens strains as A. tumefaciens, showed they differed from the DK strain. The DK strain belongs in the genus Agrobacterium, but it could not be assigned to any known species on the basis of data from phenotypic or 16S sequence comparisons. To our knowledge, this is the first report of crown gall on diffuse knapweed in the field. This strain has been deposited into the International Collection of Phytopathogenic Bacteria at Fort Detrick (Accession No. 60099), and the 16S rRNA sequence has been deposited into the GenBank database (Accession No. EF687663). References: (1) A. J. Caesar, Plant Dis. 78:796, 1994. (2) B. Holmes and P. Roberts, J. Appl. Bacteriol. 50:443, 1981. (3) L. W. Moore et al. Page 17 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (4) K. Ophel and A. Kerr, Int. J. Syst. Bacteriol. 40:236, 1990.

[A molecular-and-genetic analysis of species and strain diversity of bifidobacteria in early childhood].

The species and strain composition of bifidobacteria in 29 children both sexes, aged 8 to 16 months, was studied. Species-specific primers and PCR were used to determine to which species the predominant strains of bifidobacteria, isolated from feces by cultural methods, belonged. Bifidobacteria were found in 28 (96.5%) children; their number was 10.2 +/- 0.7 ECU per a gram of the material. B. longum and B. bifidum were frequent (71.4 and 53.5%, respectively). The level of quantitative detection used in the study also allowed revealing of B. catenulatum (17.9%) and B. breve (14.4%). A high titer of B. dentium was found in one case (3.6%). B. adolescentis and B. angulatum were not found in any patient. The average number of species found in one child was 1.7 +/- 0.7. RAPD-PCR and investigation of plasmid profile were used to determine possible belonging of the isolates to different strains. The average number of strains per one sample was 2.3 +/- 1.2. Nine unique plasmid bifidobacterial strains were isolated from 7 children. In 3 children the intestinal tract was found to be colonized by both plasmid and non-plasmid-carrying strains of one bifidobacterial species.

Genetic Characterization and Diversity of Rathayibacter toxicus.

ABSTRACT Rathayibacter toxicus is a nematode-vectored gram-positive bacterium responsible for a gumming disease of grasses and production of a highly potent animal and human toxin that is often fatal to livestock and has a history of occurring in unexpected circumstances. DNA of 22 strains of R. toxicus from Australia were characterized using amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE). AFLP analysis grouped the 22 strains into three genetic clusters that correspond to their geographic origin. The mean similarity between the three clusters was 85 to 86%. PFGE analysis generated three different banding patterns that enabled typing the strains into three genotypic groups corresponding to the same AFLP clusters. The similarity coefficient was 63 to 81% for XbaI and 79 to 84% for SpeI. AFLP and PFGE analyses exhibited an analogous level of discriminatory power and produced congruent results. PFGE analysis indicated that the R. toxicus genome was represented by a single linear chromosome, estimated to be 2.214 to 2.301 Mb. No plasmids were detected.

[Search of promising strains of bifidobacteria and lactobacillus for the development of new biopreparations]. / Poisk perspektivnykh shtammov bifidobakterii i laktobatsill dlia razrabotki novykh biopreparatov.

For the first time the species composition of bifidobacteria and lactobacilli in clinically healthy young children has been studied. As revealed in this study, the dominating species of bifidobacteria are B. longum, B. adolescentis and B. infantis, while the dominating species of lactobacilli are Lactobacillus acidophilus and L. rhamnosus. In 83 isolated cultures of bifidobacteria and 34 isolated species of lactobacilli the activity of acid formation and the antagonistic activity with respect to Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Clostridium perfringens have been tested. B. longum strain 58B, B. infantis strain 37B, L. rhamnosus strain 12L and L. acidophilus strain 27L, typical for children of this age group, having good antagonistic activity and pronounced acid-forming properties, have been selected. These strains hold good promise to be used as the basis for the development of a complex probiotic preparation for correcting intestinal microflora in young children.

[Qualitative and quantitative composition of intestinal microflora in healthy young children]. / Izuchenie kachestvennogo i kolichestvennogo sostava mikroflory kishechnika u klinicheski zdorovykh detei v rannem vozraste.

The results of the qualitative and quantitative analysis of the microflora of the large intestine in 45 healthy children aged 6, 8, 10 months and 1 year, living in Moscow, are presented. During the first year of life high concentrations and detection rate of not only bifidobacteria, enterococci, nonpathogenic Escherichia, but also bacteria, commonly regarded as opportunistic were typical. The latter include lecithinase positive Clostridium, citrate assimilating enterobacteria, as well as Escherichia with low biochemical activity and capacity for hemolysin production. In addition, this group also includes coagulase-positive staphylococci. Later on, as these children become older, opportunistic bacteria are partially or completely eliminated under normal conditions. Suggestion is made that the composition of intestinal microflora depends on protective and other systems of the host at different periods of life, as well as on the character of nutrition and the microbial contamination of the environment.