Evaluation of Trabecular Microstructure of Cancellous Bone Using Quarter-Detector Computed Tomography.

Quarter-detector computed tomography (QDCT) is an ultra-high-spatial-resolution imaging technique. This study aimed to verify the validity of trabecular structure evaluation using a QDCT scanner in the diagnosis of osteoporosis. We used a cancellous bone specimen image of the second lumbar vertebrae of an adult male with moderate osteoporosis. To obtain QDCT images, we created a three-dimensional model from micro-CT images of the specimen. Statistical analysis was performed on the relationship between micro-CT and QDCT imaging modalities. The differences between micro-CT and QDCT were assessed based on their significance with respect to the calculated mean measurements using the Mann-Whitney test. Single regression analysis was performed using linear regression, with micro-CT and QDCT as the explanatory and objective variables, respectively, to determine the relationship of the measured values between the two modalities. By applying the necessary correction to the micro-CT measured values, it is possible to perform an analysis equivalent to micro-CT, which offers higher spatial resolution than QDCT. We found evidence that if QDCT can be used, trabecular structure evaluation may contribute to image diagnosis to evaluate practical bone fragility.

Measurement of aerodynamic force and moment acting on a javelin using a magnetic suspension and balance system.

The rules governing the dimensions of the Javelin were substantially changed in 1986. It was considered that this new design guaranteed there was zero pitching moment at 0° angle of attack and that the pitching moment decreased (became negative) with increasing angle of attack. The objective of this study is to investigate if the pitching moment remains always negative (nose-down rotation). To measure accurate aerodynamic forces acting on a Javelin, the world's largest 1 m magnetic suspension and balance system was used. The magnetic suspension and balance system was able to measure aerodynamic forces without support interference in the wind tunnel. In addition, computational fluid dynamics were carried out to estimate the pitching moment coefficients. It was found that the pitching moment coefficient of a commercially available Javelin becomes positive (nose-up rotation) at lower angles of attack, less than 12°. The pitching moment becomes positive if the upstream side of the center of gravity receives more inflow than the downstream side. This situation can be attained by, for example, increasing the thickness of the upstream side when compared with that of the downstream side.

Relationship between nutritional biomarkers and occlusal status in gastric cancer patients using the Eichner index: Observational study.

ABSTRACT: Systemic inflammatory responses and nutritional status are useful prognostic factors in gastric cancer patients. Since oral hypofunction causes undernutrition, we cross-sectionally investigated whether nutritional biomarkers were affected by the occlusal supporting zone status.In 114 gastric cancer patients, the gastric cancer stage, body mass index, albumin levels, total lymphocyte counts, cholesterol levels, C-reactive protein levels, and 4 nutritional biomarkers - the Glasgow prognostic score (GPS), neutrophil-lymphocyte ratio, prognostic nutrition index (PNI), and controlling nutritional status (CONUT) - were evaluated. Oral conditions were assessed by determining the number of remaining teeth. The occlusal supporting status was based on the Eichner classification. Patients were assigned into 3 groups per their occlusal status, and mean values were compared using the Kruskal-Wallis test. The mean age and body mass index were 72.2 ± 8.5 (50-89) years and 22.0 ± 3.6 (14.8-33.4), respectively. There were 42, 39, 23, and 10 patients in stages I, II, III, and IV, respectively. The mean number of remaining teeth was 18.1 ± 9.5. According to the Eichner classification, there were 45, 42, and 27 patients in groups A, B, and C, respectively. The GPS and neutrophil-lymphocyte ratio values and CONUT frequencies between groups A and C were significantly different (P = .033, P = .00097, P = .04, respectively; Mann-Whitney U test). PNI values were lower in group C with poor occlusal support zones than in group A with stable occlusal support zones.Occlusal supporting zone reductions were undernutrition associated. Eichner Class C patients with few occlusal supporting zones had poor GPS, PNI, and CONUT values and were undernourished.

Risk factors for postoperative nausea and vomiting after the removal of impacted third molars: a cross-sectional study.

BACKGROUND: A better understanding of the risk factors for postoperative nausea and vomiting (PONV) could improve patient outcomes. This study aimed to analyze the risk factors for PONV and its onset after third molar impaction surgery, and to demonstrate the importance of controlling anesthesia-related factors regardless of patient-specific factors. METHODS: We included patients who reported nausea and vomiting within 12 h of extubation. Patients with incomplete data, those who could not communicate, and those with gastrointestinal disorders were excluded. We evaluated patient-specific risk factors, and the use of volatile anesthetics and intraoperative fentanyl anesthetic-related factors. Multiple logistic regression analysis was performed taking patient background factors into account. RESULTS: In total, 182 patients who underwent disimpaction of the third molar under general anesthesia between January 2017 and December 2018 at Nagoya Ekisaikai Hospital, were included. Approximately 12.6% (n = 23) patients experienced PONV, with no significant difference in terms of sex, smoking status, age, and body mass index compared to patients without PONV. Multiple logistic regression analysis revealed no interaction between fentanyl and volatile anesthetics. The major risk factor for PONV was the use of volatile anesthetics. Patients in whom anesthesia was maintained by volatile anesthetics were 13.35 times more likely to have PONV than those in whom total intravenous anesthesia was induced (P < 0.001). CONCLUSION: Maintenance of anesthesia with volatile anesthetics is a risk factor for PONV.

Stress-Coping Humoral Glycolipids Produced by Mice Given Controlled Bathing Treatments.

Mammalians have recognition-behavioral stress-coping neuronal module system followed by some humoral glycolipids. A sulfated Galbeta1-4GlcNAc-lipid promotes the serotonergic module regulating the emotional behaviors for not-wasting the physical strength; GalNAcalpha1-3GalNAc-lipid promotes the adrenergic module inducing the behaviors escaping from the uneasy situation, and sulfated Fucalpha1-2Gal-lipid protects the cholinergic module keeping the stressor-memory from the ischemia-stress. Mouse given bathing recognizes the stressors to be coped with in the treatment. We previously observed mouse given CO2-microbubble-bathing increased the behavior escaping from the bathing situation. Mouse given CO2-microbubble-bathing would recognize the other stressors to be coped with in the treatment. We examined stress-coping glycolipids produced by mice given controlled bathing treatments, and got the following results. A sulfated Galbeta1-4GlcNAc-lipid production was increased by the acidic bathing condition and the dissolved CO2, GalNAcalpha1-3GalNAc-lipid production was increased by the dissolved CO2, and sulfated Fucalpha1-2Gal-lipid production was increased by the acidic bathing condition. We understood the mice treated with CO2-microbubble-bathing would recognize the acidic bathing condition and the dissolved CO2, but not the microbubble, as the other stressors to be coped.

Acquisition of Quasi-Monochromatic Dual-Energy in a Microfocus X-ray Generator and Development of Applied Technology.

In regenerative medicine, evaluation of bone mineral density using a microfocus X-ray generator could eventually be used to determine the degree of bone tissue regeneration. To evaluate bone mineral density against regenerated bone material, two low-energy X-rays are necessary. Herein, the acquisition of quasi-monochromatic, dual-energy soft X-ray and the subsequent medical application were examined using the K-absorption edges of two types of metal filters (i.e., zirconium and tin) in a microfocus X-ray generator. Investigation of the optimal tube voltage and filter thickness to form a quasi-monochromatic energy spectrum with a single filter revealed that a filter thickness of 0.3 mm results in an optimal monochromatization state. When a dual filter was used, the required filter thickness was 0.3 mm for tin and 0.2 mm for zirconium at a tube voltage of 35 kV. For the medical application, we measured quasi-monochromatic, dual-energy X-rays to evaluate the measurement accuracy of bone mineral density. Using aluminum as a simulated bone sample, a relative error of ≤5% was consistent within the aluminum thickness range of 1⁻3 mm. These data suggest that a bone mineral density indicator of recycled bone material can be easily obtained with the quasi-monochromatic X-ray technique using a microfocus X-ray generator.

Evaluation of surface dose and image quality using the half-scan mode in chest computed tomography-guided interventional radiology: a phantom study.

The authors aimed to evaluate the effects of the half-scan mode on image quality and physician exposure to radiation in computed tomography (CT)-guided interventional radiology (IVR) to the right lung using an intermittent CT fluoroscopy technique for measuring phantom surface dose distribution and image noise. For the half-scan mode, settings at 0°, 90°, 180°, and 270° were used as the central axis of the X-ray exposure range on the chest phantom. With the center of the ventral side in the chest phantom defined as 0°, optically stimulated luminescent dosimeters were attached at five positions at 30° intervals on the right side of the phantom surface. Securing a space for device operation during the procedure is necessary. The couch was shifted downward by 50 mm to reproduce the conditions used for measurement in clinical settings. Image noise and contrast-to-noise ratio were measured to assess image quality; subjective evaluation was performed using simulated lung nodules placed in the phantom. The phantom surface dose distribution in the measured half-scan mode depended on the angle setting. Additionally, the phantom surface dose in the half-scan mode at the 90° setting was reduced by approximately 50%; however, image quality was clearly decreased. In CT-guided IVR to the right lung, using a lead drape and half-scan mode according to the procedural situation is important.

Evaluation of an X-Ray Dose Profile Derived from an Optically Stimulated Luminescent Dosimeter during Computed Tomographic Fluoroscopy.

The purpose of this study was to evaluate scatter radiation dose to the subject surface during X-ray computed tomography (CT) fluoroscopy using the integrated dose ratio (IDR) of an X-ray dose profile derived from an optically stimulated luminescent (OSL) dosimeter. We aimed to obtain quantitative evidence supporting the radiation protection methods used during previous CT fluoroscopy. A multislice CT scanner was used to perform this study. OSL dosimeters were placed on the top and the lateral side of the chest phantom so that the longitudinal direction of dosimeters was parallel to the orthogonal axis-to-slice plane for measurement of dose profiles in CT fluoroscopy. Measurement of fluoroscopic conditions was performed at 120 kVp and 80 kVp. Scatter radiation dose was evaluated by calculating the integrated dose determined by OSL dosimetry. The overall percent difference of the integrated doses between OSL dosimeters and ionization chamber was 5.92%. The ratio of the integrated dose of a 100-mm length area to its tails (-50 to -6 mm, 50 to 6 mm) was the lowest on the lateral side at 80 kVp and the highest on the top at 120 kVp. The IDRs for different measurement positions were larger at 120 kVp than at 80 kVp. Similarly, the IDRs for the tube voltage between the primary X-ray beam and scatter radiation was larger on the lateral side than on the top of the phantom. IDR evaluation suggested that the scatter radiation dose has a high dependence on the position and a low dependence on tube voltage relative to the primary X-ray beam for constant dose rate fluoroscopic conditions. These results provided quantitative evidence supporting the radiation protection methods used during CT fluoroscopy in previous studies.

[Fluoroscopy dose reduction of computed tomography guided chest interventional radiology using real-time iterative reconstruction].

The purpose of this study was to evaluate the radiation dose reduction to patients and radiologists in computed tomography (CT) guided examinations for the thoracic region using CT fluoroscopy. Image quality evaluation of the real-time filtered back-projection (RT-FBP) images and the real-time adaptive iterative dose reduction (RT-AIDR) images was carried out on noise and artifacts that were considered to affect the CT fluoroscopy. The image standard deviation was improved in the fluoroscopy setting with less than 30 mA on 120 kV. With regard to the evaluation of artifact visibility and the amount generated by the needle attached to the chest phantom, there was no significant difference between the RT-FBP images with 120 kV, 20 mA and the RT-AIDR images with low-dose conditions (greater than 80 kV, 30 mA and less than 120 kV, 20 mA). The results suggest that it is possible to reduce the radiation dose by approximately 34% at the maximum using RT-AIDR while maintaining image quality equivalent to the RT-FBP images with 120 V, 20 mA.

TetR-type transcriptional regulator VtpR functions as a global regulator in Vibrio tubiashii.

Vibrio tubiashii, a causative agent of severe shellfish larval disease, produces multiple extracellular proteins, including a metalloprotease (VtpA), as potential virulence factors. We previously reported that VtpA is toxic for Pacific oyster (Crassostrea gigas) larvae. In this study, we show that extracellular protease production by V. tubiashii was much reduced by elevated salt concentrations, as well as by elevated temperatures. In addition, V. tubiashii produced dramatically less protease in minimal salts medium supplemented with glucose or sucrose as the sole carbon source than with succinate. We identified a protein that belongs to the TetR family of transcriptional regulators, VtpR, which showed high homology with V. cholerae HapR. We conclude that VtpR activates VtpA production based on the following: (i) a VtpR-deficient V. tubiashii mutant did not produce extracellular proteases, (ii) the mutant showed reduced expression of a vtpA-lacZ fusion, and (iii) VtpR activated vtpA-lacZ in a V. cholerae heterologous background. Moreover, we show that VtpR activated the expression of an additional metalloprotease gene (vtpB). The deduced VtpB sequence showed high homology with a metalloprotease, VhpA, from V. harveyi. Furthermore, the vtpR mutant strain produced reduced levels of extracellular hemolysin, which is attributed to the lower expression of the V. tubiashii hemolysin genes (vthAB). The VtpR-deficient mutant also had negative effects on bacterial motility and did not demonstrate toxicity to oyster larvae. Together, these findings establish that the V. tubiashii VtpR protein functions as a global regulator controlling an array of potential virulence factors.

Virulence of metalloproteases produced by Vibrio species on Pacific oyster Crassostrea gigas larvae.

Vibrio tubiashii, a pathogen of shellfish larvae and juveniles, produces several extracellular products. Here, we document that culture supernatants of several marine Vibrio species showed toxicity to oyster larvae. Treatment of these supernatants with EDTA not only severely diminished proteolytic activities, but also dramatically reduced toxicity to the larvae. Culture supernatants of metalloprotease-deficient mutants of V. tubiashii, V. cholerae, and V. splendidus were impaired in their ability to cause larval death compared to the wild type strains. Culture supernatants of Pseudomonas aeruginosa, known to contain several secreted proteases, showed virtually no toxicity to oyster larvae. Purified V. tubiashii protease A (VtpA), but not the prototype metalloprotease, thermolysin from Bacillus thermoproteolyticus, was highly toxic to the larvae. In addition, toxicity of purified VtpA was much greater for 6-d-old oyster larvae than for 16-d-old larvae. Together, these results indicated that culture supernatants of a variety of Vibrio species are highly toxic to oyster larvae and that the production of a metalloprotease is required for this effect. We propose that there are, as yet uncharacterized, specific substrates contained in larval tissue that are degraded by VtpA as well as certain homologous metalloproteases produced by other marine Vibrio species which, in turn, may contribute to vibriosis.

The extracellular metalloprotease of Vibrio tubiashii directly inhibits its extracellular haemolysin.

Vibrio tubiashii is a re-emerging pathogen of molluscs that secretes a variety of extracellular products (ECPs), including a metalloprotease and a cytolysin/haemolysin. Previously, we reported that the V. tubiashii haemolysin locus consists of two ORFs (vthB and vthA), similar to that of the homologous haemolysin genes (vvhB and vvhA) found in Vibrio vulnificus. Here, we demonstrate that the concomitant expression of both V. tubiashii genes resulted in significantly higher haemolytic activity than the vthA gene alone. In addition, we created a VthAB- mutant strain of V. tubiashii that was virtually devoid of haemolytic activity in liquid media. Interestingly, significant production of an additional haemolysin(s) was observed on blood plates. Moreover, we have previously reported that in V. tubiashii, proteolytic and haemolytic activities are inversely produced during bacterial growth. Here, we study this correlation in more detail and present evidence that the VtpA metalloprotease inhibits haemolytic activity in culture supernatants, based on the following evidence: (i) loss of metalloprotease activity by either mutation or EDTA inhibition resulted in increased haemolytic activity; (ii) overexpression of the vtpA gene resulted in decreased haemolytic activity; (iii) purified VtpA metalloprotease directly diminished haemolytic activity by purified VthA haemolysin. Importantly, we found not only that vthAB gene expression remained high throughout growth but also that there were no dramatic differences in vthAB gene expression between the parent and VtpA- mutant strains. Thus, our results strongly suggest that the V. tubiashii metalloprotease directly targets its haemolysin.

The Rpd3/HDAC complex is present at the URS1 cis-element with hyperacetylated histone H3.

In eukaryotes, the hypoacetylated state of histone N-terminal lysines at many gene-promoters, which is created by histone deacetylases (HDACs), is changed to the hyperacetylated state by the function of histone acetyltransferases (HATs) upon transcription activation. Although much insight has been obtained to date as to how modification of the histone tail regulates gene expression, little is known about how the transition between the unmodified and modified states takes place. In Saccharomyces cerevisiae, the HDAC complex containing Rpd3 (Rpd3L) represses the transcription of several sets of genes through the URS1 cis-element. We found that the histone H3 acetylation level at the URS1 of seven genes (INO1, CAT2, ACS1, YAT1, RIM4, CRC1, and SIP4) was elevated in the presence of Rpd3/HDAC in growth in acetate-containing medium (YPA), suggesting that a mechanism that regulates HDAC activity is present in this organism. The biological significance of this phenomenon is discussed below.

Development of a quantitative real-time PCR assay for detection of Vibrio tubiashii targeting the metalloprotease gene.

Vibrio tubiashii has recently re-emerged as a pathogen of bivalve larvae, causing a marked increase in the mortality of these species within shellfish rearing facilities. This has resulted in substantial losses of seed production and thus created the need for specific as well as sensitive detection methods for this pathogen. In this project, quantitative PCR (qPCR) primers were developed and optimized based upon analysis of the V. tubiashii vtpA gene sequence, encoding a metalloprotease known to cause larval mortality. Standard curves were developed utilizing dilutions of known quantities of V. tubiashii cells that were compared to colony forming unit (CFU) plate counts. The assay was optimized for detection of vtpA with both lab-grown V. tubiashii samples and filter-captured environmental seawater samples seeded with V. tubiashii. In addition, the primers were confirmed to specifically detect only V. tubiashii when tested against a variety of non-target Vibrio species. Validation of the assay was completed by analyzing samples obtained from a shellfish hatchery. The development of this rapid and sensitive assay for quantitative detection of V. tubiashii will accurately determine levels of this bacterium in a variety of seawater samples, providing a useful tool for oyster hatcheries and a method to assess the presence of this bacterium in the current turbulent ocean environment.

The extracellular metalloprotease of Vibrio tubiashii is a major virulence factor for pacific oyster (Crassostrea gigas) larvae.

Vibrio tubiashii is a recently reemerging pathogen of larval bivalve mollusks, causing both toxigenic and invasive disease. Marine Vibrio spp. produce an array of extracellular products as potential pathogenicity factors. Culture supernatants of V. tubiashii have been shown to be toxic to oyster larvae and were reported to contain a metalloprotease and a cytolysin/hemolysin. However, the structural genes responsible for these proteins have yet to be identified, and it is uncertain which extracellular products play a role in pathogenicity. We investigated the effects of the metalloprotease and hemolysin secreted by V. tubiashii on its ability to kill Pacific oyster (Crassostrea gigas) larvae. While V. tubiashii supernatants treated with metalloprotease inhibitors severely reduced the toxicity to oyster larvae, inhibition of the hemolytic activity did not affect larval toxicity. We identified structural genes of V. tubiashii encoding a metalloprotease (vtpA) and a hemolysin (vthA). Sequence analyses revealed that VtpA shared high homology with metalloproteases from a variety of Vibrio species, while VthA showed high homology only to the cytolysin/hemolysin of Vibrio vulnificus. Compared to the wild-type strain, a VtpA mutant of V. tubiashii not only produced reduced amounts of protease but also showed decreased toxicity to C. gigas larvae. Vibrio cholerae strains carrying the vtpA or vthA gene successfully secreted the heterologous protein. Culture supernatants of V. cholerae carrying vtpA but not vthA were highly toxic to Pacific oyster larvae. Together, these results suggest that the V. tubiashii extracellular metalloprotease is important in its pathogenicity to C. gigas larvae.

Re-emergence of Vibrio tubiashii in bivalve shellfish aquaculture: severity, environmental drivers, geographic extent and management.

During 2006 and 2007, we documented the re-emergence of severe episodes of vibriosis caused by Vibrio tubiashii in shellfish hatcheries on the west coast of North America. Lost larval and juvenile production included 3 previously undescribed hosts, Pacific (Crassostrea gigas) and Kumamoto (C. sikamea) oysters and geoduck clams Panope abrupta, with a 2007 decline in larval oyster production of approximately 59% in one hatchery. Losses of larval and juvenile bivalves were linked to V. tubiashii blooms in the coastal environment, which were associated with the apparent mixing of unusually warm surface seawater and intermittently upwelled cooler, nutrient- and Vibrio spp.- enriched seawater. The ocean temperature elevation anomaly in 2007 was not clearly linked to an El Niño event, as was a similar episode in 1998. Concentrations of the dominant shellfish-pathogenic vibrios were as high as 1.6 x 10(5) cfu ml(-1) in the cold, upwelled water. The bacteria possessed the genes coding for a protease and hemolysin described for V. tubiashii, and pathogenic isolates secreted these peptides. Lesions resulting from a classic invasive disease and a toxigenic noninvasive disease occurred in oyster and geoduck clam larvae. Management and prevention require reduction of incoming concentrations of the bacteria, reduction of contamination in water and air supplies and in stock chemical solutions, removal of bacterial toxins, and interruption of the cycle of bacterial amplification in the hatchery and in microalgal food supplies.

Bilaterally existing sternoclavicularis anticus muscles.

A 92-year-old Japanese woman was found to have the sternoclavicularis anticus muscles existing bilaterally during the course of educational dissection at Nagoya City University Medical School. It was confirmed that these muscles received the nerve supply from small branches of the lateral pectoral nerves from the brachial plexus. Herein is documented the precise gross anatomical findings with some morphometric measurements. Moreover, the morphology of this muscle is discussed in relation to previously described variations of the pectoral sheet of muscles.

[Examination of tumor diameter measurement precision by RECIST].

Image evaluation with Response Evaluation Criteria in Solid Tumors (RECIST) evaluates the change in a measurable lesion as determined by ruler or micrometer caliper. However, there is no definition of the conditions thought to influence the precision of measurement. We therefore examined the effects on measurement precision by changing image amplification, WW, WL, and time phase. Moreover, to determine response rate, one-dimensional evaluation with RECIST was compared with the two-dimensional evaluation of World Health Organization (WHO) for a hepatocellular carcinoma. The results of measuring the object lesion for measured value variation were as follows. Under image conditions of 1 time expansion/ WW 150/ WL 100 was (4.92+/-1.94)%. Under image conditions of 1 time expansion/ WW 350/ WL 75 was (4.42+/-1.70)%. Under image conditions of 4 times expansion/ WW 150/ WL 100 was (2.52+/-0.82)%. Under image conditions of 4 times expansion/ WW 350/ WL 75 was (2.83+/-1.10)%. When an image was enlarged to 4 times, precision doubled. There was no a difference in comparing RECIST to WHO in terms of response rate. Thus the best method was considered to be RECIST because of its convenience.

PsrA, the Pseudomonas sigma regulator, controls regulators of epiphytic fitness, quorum-sensing signals, and plant interactions in Pseudomonas syringae pv. tomato strain DC3000.

Pseudomonas syringae pv. tomato strain DC3000, a pathogen of tomato and Arabidopsis, occurs as an epiphyte. It produces N-acyl homoserine lactones (AHLs) which apparently function as quorum-sensing signals. A Tn5 insertion mutant of DC3000, designated PsrA(-) (Psr is for Pseudomonas sigma regulator), overexpresses psyR (a LuxR-type regulator of psyI) and psyI (the gene for AHL synthase), and it produces a ca. 8-fold-higher level of AHL than does DC3000. The mutant is impaired in its ability to elicit the hypersensitive reaction and is attenuated in its virulence in tomato. These phenotypes correlate with reduced expression of hrpL, the gene for an alternate sigma factor, as well as several hrp and hop genes during early stages of incubation in a Hrp-inducing medium. PsrA also positively controls rpoS, the gene for an alternate sigma factor known to control various stress responses. By contrast, PsrA negatively regulates rsmA1, an RNA-binding protein gene known to function as negative regulator, and aefR, a tetR-like gene known to control AHL production and epiphytic fitness in P. syringae pv. syringae. Gel mobility shift assays and other lines of evidence demonstrate a direct interaction of PsrA protein with rpoS promoter DNA and aefR operator DNA. In addition, PsrA negatively autoregulates and binds the psrA operator. In an AefR(-) mutant, the expression of psyR and psyI and AHL production are lower than those in DC3000, the AefR(+) parent. In an RpoS(-) mutant, on the other hand, the levels of AHL and transcripts of psyR and psyI are much higher than those in the RpoS(+) parent, DC3000. We present evidence, albeit indirect, that the RpoS effect occurs via psyR. Thus, AefR positively regulates AHL production, whereas RpoS has a strong negative effect. We show that AefR and RpoS do not regulate PsrA and that the PsrA effect on AHL production is exerted via its cumulative, but independent, effects on both AefR and RpoS.

Erwinia carotovora subspecies produce duplicate variants of ExpR, LuxR homologs that activate rsmA transcription but differ in their interactions with N-acylhomoserine lactone signals.

The N-acylhomoserine lactone (AHL) signaling system comprises a producing system that includes acylhomoserine synthase (AhlI, a LuxI homolog) and a receptor, generally a LuxR homolog. AHL controls exoprotein production in Erwinia carotovora and consequently the virulence for plants. In previous studies we showed that ExpR, a LuxR homolog, is an AHL receptor and that it activates transcription of rsmA, the gene encoding an RNA binding protein which is a global negative regulator of exoproteins and secondary metabolites. An unusual finding was that the transcriptional activity of ExpR was neutralized by AHL. We subsequently determined that the genomes of most strains of E. carotovora subspecies tested possess two copies of the expR gene: expR1, which was previously studied, and expR2, which was the focus of this study. Comparative analysis of the two ExpR variants of E. carotovora subsp. carotovora showed that while both variants activated rsmA transcription, there were significant differences in the patterns of their AHL interactions, the rsmA sequences to which they bound, and their relative efficiencies of activation of rsmA transcription. An ExpR2- mutant produced high levels of exoproteins and reduced levels of RsmA in the absence of AHL. This contrasts with the almost complete inhibition of exoprotein production and the high levels of RsmA production in an AhlI- mutant that was ExpR1-. Our results suggest that ExpR2 activity is responsible for regulating exoprotein production primarily by modulating the levels of an RNA binding protein.