Revascularization in Immature and Mature Teeth with Necrotic Pulp: A Clinical Study.

AIM: The aim of this in vivo study was to evaluate the revascularization procedure both in immature and mature teeth with necrotic pulp and open apices, disinfected with triple antibiotic paste followed by inducing blood clot in the root canal. MATERIALS AND METHODS: Fifteen patients were selected who presented with immature and mature permanent teeth with pulpal necrosis and open apices. In the first visit, the root canal was accessed with LA and rubber dam isolation; then the canal was disinfected using triple antibiotic paste containing ciprofloxacin, metronidazole, and clindamycin in the ratio of 1:1:1 and closed with IRM. In the second visit, after administering local anesthesia and isolating with a rubber dam, the triple antibiotic paste was washed out by saline irrigation, and apical papilla beyond the confines of the root canal was stimulated with sterile H file to produce a blood clot. Finally, the access was closed using a double seal with mineral trioxide aggregate (MTA) placed apical to cementoenamel junction and resin bonded cement over the MTA. Radiographic examination and pulp sensibility test was done during the follow-up period of 2,4,6,8 and 10 months. RESULT: After 10 months follow-up, 10 out of 13 patients showed root development and apical closure. The eight patients out of 13 showed root development, apical closure and lateral thickening of radicular dentin and 2 out of 13 patients showed a positive response to electric sensibility test. CONCLUSION: Within the limitation of this study, it can be concluded that there is evidence of root development, increase in lateral wall thickness, apical closure and positive response to pulp sensibility test in both mature and immature teeth with necrotic pulp. CLINICAL SIGNIFICANCE: The conventional approach for management of teeth with necrotic pulp and the open apex is altered with the possibility of tissue regeneration within the pulp space and continued root development through revascularization procedures. It also re-establishes the vitality in a previously nonvital and necrosed tooth.

Evolution of the metabolic and regulatory networks associated with oxygen availability in two phytopathogenic enterobacteria.

BACKGROUND: Dickeya dadantii and Pectobacterium atrosepticum are phytopathogenic enterobacteria capable of facultative anaerobic growth in a wide range of O2 concentrations found in plant and natural environments. The transcriptional response to O2 remains under-explored for these and other phytopathogenic enterobacteria although it has been well characterized for animal-associated genera including Escherichia coli and Salmonella enterica. Knowledge of the extent of conservation of the transcriptional response across orthologous genes in more distantly related species is useful to identify rates and patterns of regulon evolution. Evolutionary events such as loss and acquisition of genes by lateral transfer events along each evolutionary branch results in lineage-specific genes, some of which may have been subsequently incorporated into the O2-responsive stimulon. Here we present a comparison of transcriptional profiles measured using densely tiled oligonucleotide arrays for two phytopathogens, Dickeya dadantii 3937 and Pectobacterium atrosepticum SCRI1043, grown to mid-log phase in MOPS minimal medium (0.1% glucose) with and without O2. RESULTS: More than 7% of the genes of each phytopathogen are differentially expressed with greater than 3-fold changes under anaerobic conditions. In addition to anaerobic metabolism genes, the O2 responsive stimulon includes a variety of virulence and pathogenicity-genes. Few of these genes overlap with orthologous genes in the anaerobic stimulon of E. coli. We define these as the conserved core, in which the transcriptional pattern as well as genetic architecture are well preserved. This conserved core includes previously described anaerobic metabolic pathways such as fermentation. Other components of the anaerobic stimulon show variation in genetic content, genome architecture and regulation. Notably formate metabolism, nitrate/nitrite metabolism, and fermentative butanediol production, differ between E. coli and the phytopathogens. Surprisingly, the overlap of the anaerobic stimulon between the phytopathogens is also relatively small considering that they are closely related, occupy similar niches and employ similar strategies to cause disease. There are cases of interesting divergences in the pattern of transcription of genes between Dickeya and Pectobacterium for virulence-associated subsystems including the type VI secretion system (T6SS), suggesting that fine-tuning of the stimulon impacts interaction with plants or competing microbes. CONCLUSIONS: The small number of genes (an even smaller number if we consider operons) comprising the conserved core transcriptional response to O2 limitation demonstrates the extent of regulatory divergence prevalent in the Enterobacteriaceae. Our orthology-driven comparative transcriptomics approach indicates that the adaptive response in the eneterobacteria is a result of interaction of core (regulators) and lineage-specific (structural and regulatory) genes. Our subsystems based approach reveals that similar phenotypic outcomes are sometimes achieved by each organism using different genes and regulatory strategies.

Transcriptome changes associated with anaerobic growth in Yersinia intermedia (ATCC29909).

BACKGROUND: The yersiniae (Enterobacteriaceae) occupy a variety of niches, including some in human and flea hosts. Metabolic adaptations of the yersiniae, which contribute to their success in these specialized environments, remain largely unknown. We report results of an investigation of the transcriptome under aerobic and anaerobic conditions for Y. intermedia, a non-pathogenic member of the genus that has been used as a research surrogate for Y. pestis. Y. intermedia shares characteristics of pathogenic yersiniae, but is not known to cause disease in humans. Oxygen restriction is an important environmental stimulus experienced by many bacteria during their life-cycles and greatly influences their survival in specific environments. How oxygen availability affects physiology in the yersiniae is of importance in their life cycles but has not been extensively characterized. METHODOLOGY/PRINCIPAL FINDINGS: Tiled oligonucleotide arrays based on a draft genome sequence of Y. intermedia were used in transcript profiling experiments to identify genes that change expression in response to oxygen availability during growth in minimal media with glucose. The expression of more than 400 genes, constituting about 10% of the genome, was significantly altered due to oxygen-limitation in early log phase under these conditions. Broad functional categorization indicated that, in addition to genes involved in central metabolism, genes involved in adaptation to stress and genes likely involved with host interactions were affected by oxygen-availability. Notable among these, were genes encoding functions for motility, chemotaxis and biosynthesis of cobalamin, which were up-regulated and those for iron/heme utilization, methionine metabolism and urease, which were down-regulated. CONCLUSIONS/SIGNIFICANCE: This is the first transcriptome analysis of a non-pathogenic Yersinia spp. and one of few elucidating the global response to oxygen limitation for any of the yersiniae. Thus this study lays the foundation for further experimental characterization of oxygen-responsive genes and pathways in this ecologically diverse genus.