Quorum sensing in the plant pathogen Erwinia carotovora subsp. carotovora: the role of expR(Ecc).

The production of the main virulence determinants of the plant pathogen Erwinia carotovora subsp. carotovora, the extracellular cell wall-degrading enzymes, is partly controlled by the diffusible signal molecule N-(3-oxohexanoyl)-L-homoserine lactone (OHHL). OHHL is synthesized by the product of the expI/carI gene. Linked to expI we found a gene encoding a putative transcriptional regulator of the LuxR-family. This gene, expR(Ecc), is transcribed convergently to the expI gene and the two open reading frames are partially overlapping. The ExpR(Ecc) protein showed extensive amino acid sequence similarity to the repressor EsaR from Pantoea stewartii subsp. stewartii (formerly Erwinia stewartii subsp. stewartii) and to the ExpR(Ech) protein of Erwinia chrysanthemi. Inactivation of the E. carotovora subsp. carotovora expR(Ecc) gene caused no decrease in virulence or production of virulence determinants in vitro. In contrast, there was a slight increase in the maceration capacity of the mutant strain. The effects of ExpR(Ecc) were probably mediated by changes in OHHL levels. Inactivation of expR(Ecc) resulted in increased OHHL levels during early logarithmic growth. In addition, overexpression of expR(Ecc) caused a clear decrease in the production of virulence determinants and part of this effect was likely to be caused by OHHL binding to ExpR(Ecc). ExpR(Ecc) did not appear to exhibit transcriptional regulation of expI, but the effect on OHHL was apparently due to other mechanisms.

A two-component regulatory system, pehR-pehS, controls endopolygalacturonase production and virulence in the plant pathogen Erwinia carotovora subsp. carotovora.

Genes coding for the main virulence determinants of the plant pathogen Erwinia carotovora subsp. carotovora, the plant cell wall-degrading enzymes, are under the coordinate control of global regulator systems including both positive and negative factors. In addition to this global control, some virulence determinants are subject to specific regulation. We have previously shown that mutations in the pehR locus result in reduced virulence and impaired production of one of these enzymes, an endopolygalacturonase (PehA). In contrast, these pehR strains produce essentially wild-type levels of other extracellular enzymes including pectate lyases and cellulases. In this work, we characterized the pehR locus and showed that the DNA sequence is composed of two genes, designated pehR and pehS, present in an operon. Mutations in either pehR or pehS caused a Peh-negative phenotype and resulted in reduced virulence on tobacco seedlings. Complementation experiments indicated that both genes are required for transcriptional activation of the endopolygalacturonase gene, pehA, as well as restoration of virulence. Structural characterization of the pehR-pehS operon demonstrated that the corresponding polypeptides are highly similar to the two-component transcriptional regulators PhoP-PhoQ of both Escherichia coli and Salmonella typhimurium. Functional similarity of PehR-PehS with PhoP-PhoQ of E. coli and S. typhimurium was demonstrated by genetic complementation.

Two-component regulators involved in the global control of virulence in Erwinia carotovora subsp. carotovora.

Production of extracellular, plant cell wall degrading enzymes, the main virulence determinants of the plant pathogen Erwinia carotovora subsp. carotovora, is coordinately controlled by a complex regulatory network. Insertion mutants in the exp (extracellular enzyme production) loci exhibit pleiotropic defects in virulence and the growth-phase-dependent transcriptional activation of genes encoding extracellular enzymes. Two new exp mutations, designated expA and expS, were characterized. Introduction of the corresponding wild-type alleles to the mutants complemented both the lack of virulence and the impaired production of plant cell wall degrading enzymes. The expA gene was shown to encode a 24-kDa polypeptide that is structurally and functionally related to the uvrY gene product of Escherichia coli and the GacA response regulator of Pseudomonas fluorescens. Functional similarity of expA and uvrY was demonstrated by genetic complementation. The expA gene is organized in an operon together with a uvrC-like gene, identical to the organization of uvrY and uvrC in E. coli. The unlinked expS gene encodes a putative sensor kinase that shows 92% identity to the recently described rpfA gene product from another E. carotovora subsp. carotovora strain. Our data suggest that ExpS and ExpA are members of two-component sensor kinase and response regulator families, respectively. These two proteins might interact in controlling virulence gene expression in E. carotovora subsp. carotovora.

Histologic investigations on 33 retrieved Nobelpharma implants.

Thirty Nobelpharma implants were retrieved from 17 patients despite a remaining clinical stability, after between 1 and 16 years of clinical function. The reasons for implant removal were bone resorption in combination with soft tissue disorders, psychological causes, implant fracture and post mortem cases. When measured at the cortical passage, there was an average of 84.9% direct bone-to-implant contact and 81.8% average surface bone area in individual threads as evaluated in a computerized morphometric system at the light microscopic level.

Bone repair in implant models: a review with emphasis on the harvest chamber for bone regeneration studies.

The vital microscopic chamber is an experimental implant of commercially pure titanium that admits in vivo and in situ observations of bone vascularity and bone remodeling. The bone growth chamber and the bone harvest chamber, in particular, are useful tools for quantifying bone regeneration under the most variable experimental conditions.

Osseointegrated oral implants. A Swedish multicenter study of 8139 consecutively inserted Nobelpharma implants.

Fourteen Swedish teams outside the University of Gothenburg, each with minimally three years' experience in the Nobelpharma osseointegrated implant participated in a retrospective multiclinic study. The total number of consecutively inserted implants at the 14 clinics was 8139. The outcome of every implant was reported and all implant failures, irrespective of when they occurred, were published. The success criteria included absence of implant mobility, absence of radiolucent zones on x-rays, and an annual bone loss after the first year of less than 0.2 mm. In the mandible 334 implants were followed for five to eight years, with only three failures, for a success rate of 99.1%. In the maxilla 106 implants were followed for five to seven years, with a success rate of 84.9%. In irradiated and grafted mandibles, 56 implants were inserted and none was lost during a follow-up of up to five years. In the irradiated maxilla there were 16 implants inserted with three reported failures and in the grafted upper jaw 71 implants were inserted with 12 failures. The proportions of mandibular and maxillary sleeping implants were 0.8 and 0.3%, of patient drop-out implants 0.3 and 0.6%, and of patient death implants 0.9 and 1.2%, respectively. It was concluded that the osseointegrated implant, if inserted according to the guidelines of Brånemark, results in a very high degree of clinical success, thereby meeting any published oral implant success criteria.

Heat caused by drilling cortical bone. Temperature measured in vivo in patients and animals.

In vivo temperature measurements were performed at drilling in the femoral cortex of the rabbit, dog and man. In the clinical study the bone temperature was measured at fixation of a Richards plate to stabilize a pertrochanteric fracture. With a drill speed of around 20 000 rpm and saline cooling, temperatures of 40 degrees C in rabbits, 56 degrees C in dogs and 89 degrees C in patients were recorded at a distance of 0.5 mm from the periphery of the drill hole. The difference in temperature between the animal and clinical studies was mainly attributed to the difference in cortical thickness between the species. When drilling straight through the canine femur from the lateral to the medial side, a 9 degrees C higher temperature was measured in the remote, medial cortex compared to that recorded in the lateral cortex. This difference arose because it is difficult for the cooling agent to reach the medial cortex. The results of the present study indicate that temperatures measured in animal experiments are not applicable to the clinical situation where very high temperatures may arise on drilling in cortical bone, even if saline cooling is used.