Inactivation of the lpcC gene alters surface-related properties and symbiotic capability of Bradyrhizobium japonicum.

UNLABELLED: We investigated the role of the Bradyrhizobium japonicum lpcC gene, encoding a mannosyl transferase, involved in the lipopolysaccharide (LPS) biosynthesis. The inactivation of the lpcC gene considerably altered the LPS structure and the cell surface properties. LPS analysis showed that the lpcC mutant JS715 had an abnormal LPS structure deficient in O-antigen. The cell surface hydrophobicity increased approximately threefold in JS715 compared to the wild type. The increased cell surface hydrophobicity is likely to be related with cell aggregation in the mutant culture. For the growth comparison, JS715 showed slower growth rate than the wild type. The motility of JS715 decreased in soft agar plates, but it showed enhanced biofilm-forming ability. Interestingly, JS715 was not able to nodulate the host legume soybean (Glycine max). This study shows not only that lpcC is involved in the biosynthesis of O-antigen in the B. japonicum LPS, but also that inactivation of the lpcC gene affects symbiotic capability of B. japonicum and surface-related properties such as cell hydrophobicity, biofilm formation and motility. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the role of the B. japonicum lpcC in nodulation with soybean and importance of cell surface hydrophobicity. The results also highlight that intact LPS is required for successful symbiosis between B. japonicum and soybeans. Our findings not only support previous studies emphasizing the necessity of LPS on the interaction between the two symbiotic partners, but also contribute to a better understanding of the symbiotic mechanisms.

Advances in the management of UVR-associated skin cancers: autoimmune diseases and UV protection.

Ultra violet radiation (UVR) is an important feature for the development or aggravation of several dermatologic diseases. In autoimmune skin diseases it has been suggested as an important cofactor in autoimmune bullous skin diseases and more importantly cutaneous lupus erythematosus (CLE). The pathophysiological role of UVR in CLE is a result of several effects which are triggered by UVR. In detail UVR induces apoptosis of keratinocytes and an abnormal local immune response which triggers inflammation in the skin. These findings result in the clinical approach of a stringent UVR protection in affected patients. Currently UVR protection is advised to patients as a supportive measure but cannot be prescribed to patients as these products are not licensed. Well-defined prospective placebo controlled studies regarding UVR protection are missing.

Susceptibility of porcine keratocytes to immune-mediated damage in xeno-related rejection.

We admixed cultured porcine keratocytes or corneal endothelial cells in the presence of human sera or peripheral blood mononuclear cells (PBMCs) for 4 to 72 hours to investigate their immune-related susceptibilities to xeno-related rejection. We evaluated complement deposition at 48 hours by flow cytometry after staining with the C3 anti-goat cy3 antibody. The inhibition of proliferation of porcine corneal cells by human sera was examined using the 3-[4,5-dimethy/thiazol-2,5-dephenyl tetrazolium bromide (MTT) assay over 24 to 72 hours. The amount of 51chromium (Cr)-release was estimated after a reaction between the porcine cells and human PBMCs for 4 hours. There was greater C3 deposition in keratocytes (60.2%) than in endothelial cells (26.9%; P = .05, Mann-Whitney U test). Both keratocytes and endothelial cells showed significant levels of proliferative inhibition over a period of 72 hours. The number of 51Cr-release cells on interleukin-2 addition was significantly higher among keratocytes (88.0%) than endothelial cells (51.4%) at a 1:100 target:effector ratio (P = .04, Mann-Whitney U test). Our present data suggested that porcine keratocytes might be key target cells in xeno-related rejections when the porcine cornea is transplanted to primates.

Is a short arm cast appropriate for stable distal radius fractures in patients older than 55 years? A randomized prospective multicentre study.

We conducted a prospective randomized, multicentre study to compare short arm and long arm plaster casts for the treatment of stable distal radius fracture in patients older than 55 years. We randomly assigned patients over the age of 55 years who had stable distal radius fracture to either a short arm or long arm plaster cast at the first review 1 week after their injury. Radiographic and clinical follow-up was conducted at 1, 3, 5, 12 and 24 weeks following their injury. Also, degree of disability caused by each cast immobilization was evaluated at the patient's visit to remove the cast. There were no significant differences in radiological parameters between the groups except for volar tilt. Despite these differences in volar tilt, neither functional status as measured by the Disabilities of the Arm, Shoulder and Hand, nor visual analogue scale was significantly different between the groups. However, the mean score of disability caused by plaster cast immobilization and the incidence rate of shoulder pain were significantly higher in patients who had a long plaster cast. Our findings suggest that a short arm cast is as effective as a long arm cast for stable distal radius fractures in the elderly. Furthermore, it is more comfortable and introduces less restriction on daily activities. LEVEL OF EVIDENCE: II.

Time series study of a 17-year record of (7)Be and (210)Pb fluxes in northern Taiwan using ensemble empirical mode decomposition.

Using the ensemble empirical mode decomposition (EEMD) and a significance test method, we have analyzed time series data on the fluxes of (7)Be and (210)Pb collected over a span of 17 y in Northern Taiwan. Among nine intrinsic mode functions (IMFs) extracted from the method five (IMF4-8) are non-trivial for (210)Pb and have adequate S/N with significant power in localized windows around the periodicities of 0.5 y, 1 y, 2 y, 5 y, and 11 y, respectively. For (7)Be, IMF5 and IMF8 with periods around 1 y and 11 y, respectively, have adequate S/N. The semi-annual and annual cycles represented by IMF4 and IMF5, respectively, are dominated by East Asian monsoon. The sum of IMF6 and IMF7 reveals an inter-annual cycle where both (7)Be and (210)Pb fluxes are well-correlated with the East Asian winter monsoon index (EAWMI). The close tracking of the (210)Pb and (7)Be in IMF8 cases may reflect an 11 y cycle; implying that it is caused by common climatologic factors, likely related to solar cycle, rather than their distinct production modes.

[4 + 3] Cycloaddition of cyclopropanone hemiacetals.

[reaction: see text]. Intermolecular and intramolecular [4 + 3] cycloaddition reactions of readily available cyclopropanone hemiacetals with furans are described.

Facile synthesis of the tricyclic core of sarain A. 3-Oxidopyridinium betaine cycloaddition approach.

[formula: see text] A new approach to a suitably functionalized tricyclic core of sarains has been developed by means of Katritzky's cycloaddition using 3-oxidopyridinium betaines. A key step was the regioselective differentiation of the two nearly identical hydroxy groups derived from oxidative cleavage of the double bond in 8 to afford 14. A stereocontrolled construction of the tricyclic core 20 of sarains containing the requisite side chain at C-3' was achieved by an intramolecular conjugate addition.

General analysis of (14)N (I = 1) electron spin echo envelope modulation.

The analysis methods described to date for (14)N electron spin echo envelope modulation (ESEEM) mostly deal with isotropic g- and (14)N hyperfine coupling tensors. However, many cases of rhombic tensors are encountered. In the present report we present general equations for analyzing orientation-selective ESEEM and illustrate their use. (i) We present general equations for the nuclear interactions in an electron spin system where the EPR signal arises from an isolated Kramers doublet, then give the nuclear (electron-nuclear double resonance) frequencies for I = 1 associated with such a system. (ii) These are incorporated into equations for single-crystal ESEEM amplitudes, which in turn are incorporated into general equations for the orientation-selective ESEEM that arises when the EPR envelope of a frozen-solution (powder) sample is determined by g anisotropy. (iii) This development is first used in the simplest limit of an isotropic g-tensor and leads to a more general picture of the response of the I = 1 modulation amplitude to variations in the nuclear hyperfine and quadrupole coupling constants, relative to the nuclear Zeeman interaction, than had been presented previously. We find that strong modulation occurs not only in the well-known regime where the "exact/near cancellation" condition (A/2 approximately nu(N)) is satisfied, but also when the nuclear hyperfine interaction is much larger than the nuclear Zeeman interaction (A/nu(N) > 3) with A/K = 4 approximately 5. (iv) We then describe the orientation-selective (14)N ESEEM frequency-domain patterns (g vs frequency) in the presence of anisotropic (rhombic) hyperfine and electron Zeeman interactions for both coaxial and noncoaxial cases. We derive analytical solutions when the g-, hyperfine, and nuclear quadrupole tensors are coaxial. (v) The method is applied to the ESEEM of the nitrogenase MoFe protein (Av1) to determine the full hyperfine and nuclear quadrupole tensors of (14)N nuclei interacting with the S = 32 FeMo-cofactor (Fe(7)S(8)Mo: homocitrate).

Interfacial reaction and its influence on the mechanical properties of the ASZ short fibre-reinforced Al alloy composite.

The effect of interfacial reaction on the mechanical properties of the AC8A Al alloy reinforced with ASZ short fibres (ASZ/AC8A composite) was studied. In the ASZ/AC8A composite, the interfacial reaction was observed to proceed between the SiO2 binder layer and Mg of the matrix to form MgAl2O4 at the interface. Formation of MgAl2O4 was believed to enhance the interfacial bonding strength, resulting in the improved composite strength. However, the interfacial reaction in the ASZ/AC8A composite always took place at the expense of Mg in the matrix, resulting in the composite devoid of the Mg bearing precipitates such as Al2CuMg and Mg2Si. Interfacial reaction mechanisms were investigated for composites containing various Mg contents. The resultant mechanical properties of the composite were measured to determine the adequate amount of Mg within the composite. Microstructural changes of the composite were observed using transmission electron microscopy and differential scanning calorimetry to provide qualitative analyses on the experimental observations.

Glucosylation of Salicylic Acid in Nicotiana tabacum Cv. Xanthi-nc.

ABSTRACT Salicylic acid (SA) is a key regulatory component of disease resistance in plants. In tobacco mosaic virus (TMV)-inoculated tobacco (Nicotiana tabacum cv. Xanthi-nc NN genotype), newly synthesized SA is converted primarily to SA 2-O-beta-D-glucoside (SAG) and glucosyl salicylate (GS), a relatively minor metabolite. Similar patterns in the formation of GS and SAG were observed in tobacco inoculated with Pseudomonas syringae pv. phaseolicola, suggesting the accumulation of two glucosylated metabolites is a general phenomenon in tobacco plants. After SA infiltration, GS was synthesized rapidly, reached a maximal level at 6 h, declined, and remained relatively constant for at least 24 h. In contrast, SAG content increased gradually after SA treatment. Our in vitro and in vivo data suggest that a high concentration of free SA triggers transient formation of GS and continuous accumulation of SAG, which is a more stable metabolite of SA. The two distinct SA glucosyltransferases catalyzed the formation of GS and SAG, respectively. The activities of these enzymes were enhanced by TMV or P. syringae pv. phaseolicola inoculation or SA treatment and were found in different fractions of gel filtration chromatography.

Prohevein is poorly processed but shows enhanced resistance to a chitin-binding fungus in transgenic tomato plants.

In latex of rubber tree (Hevea brasiliensis), prohevein, homologous to potato win gene-encoded proteins, is processed to yield mature hevein. This mature hevein is composed of one chitin-binding domain and the C-terminal polypeptide homologous to pathogenesis-related proteins such as tobacco PR-4 and tomato P2 proteins. In contrast, prohevein was poorly cleaved to form the C-terminal polypeptide in transgenic tomato plants expressing hevein gene (HEV1)-driven polypeptides. However, mature hevein, the N-terminal cleavage form, was not found in this system. Immunoblot analysis of extracellular and intracellular fluid proteins showed that HEV1-encoded polypeptides accumulated intracellularly. In addition, retardation of growth of Trichoderma hamatum was observed in transgenic tomatoes constitutively expressing HEV1-encoded proteins.

Prediction of common bile duct stones: its validation in laparoscopic cholecystectomy.

BACKGROUND/AIMS: Although perioperative cholangiography is valuable and highly accurate in the detection of common bile duct (CBD) stones, its routine use is controversial, particularly in the era of the laparoscopic cholecystectomy because of its inherent disadvantages. The purposes of this retrospective and prospective study on cholelithiasis were to identify patients at low risk for CBD stones and to assess the validity of the low risk criteria. METHODOLOGY: For the first, retrospective study, 15 significant preoperative clinical, biochemical and sonographic variables were selected from 561 consecutive patients who underwent conventional cholecystectomy with routine intraoperative cholangiography (IOC) for cholelithiasis from January 1985 to December 1993, and independent risk factors predicting the presence of CBD stones were determined by multivariate logistic regression analysis. For the second, prospective study, from April 1994 to September 1995, a laparoscopic cholecystectomy (LC) was performed without perioperative cholangiography in 153 consecutive patients with the primary low risk criterion (sonographic CBD diameter < 10 mm) determined by the first study. All of the LC patients were followed-up for a median duration of 12 months (range 4 to 21 months). RESULTS: In the first study, CBD stones were present in 95 (16.9%) patients. The most important independent predictor was a dilated CBD (> 10 mm). Three levels of risk were determined: (1) the low risk group (73.8% of the patients), in which the CBD was not dilated and the prevalence of CBD stones was 1.5% (6/408); the moderate risk group (7.8% of the patients), in which there was a dilated CBD with normal liver function tests and a prevalence of stones of 48.8% (21/43); and the high risk group (18.4%), in which there was a dilated CBD and abnormal liver function tests and a prevalence of stones of 66.7% (68/102). In the second study, two cases (1.4%) of symptomatic overlooked CBD stones were found on endoscopic retrograde cholangiography and retrieved by endoscopic sphincterotomy on postoperative days 18 and 20, respectively. CONCLUSIONS: Preoperative assessment in cases of cholelithiasis can determine which patients are at low risk for having CBD stones, thereby avoiding unnecessary perioperative cholangiography. This selectivity is also valid in LC, since the incidence of symptomatic, overlooked CBD stones was very low.

Light, carbon dioxide, and octenol-baited mosquito trap and host-seeking activity evaluations for mosquitoes in a malarious area of the Republic of Korea.

Two field trials for commercially available and experimental mosquito traps variously baited with light, carbon dioxide, octenol, or combinations of these were evaluated in a malarious area at Paekyeon-Ri near Tongil-Chon (village) and Camp Greaves, Paju County, Kyonggi Province, Republic of Korea. The host-seeking activity for common mosquito species was determined using hourly aspirator collections from a human- and propane lantern-baited Shannon trap. The total number of mosquitoes and number of each species captured during the test were compared using 8 x 8 and 5 x 5 Latin square designs based on trap location. Significant differences were observed for the total number of mosquitoes collected in the 8 x 8 test, such that counterflow geometry (CFG) with CO2 > or = CFG with CO2 and octenol > or = Shannon trap > or = Mosquito Magnet with octenol > American Biophysics Corporation (ABC) light trap with light, CO2 (500 ml/min), and octenol > or = ABC light trap with light and dry ice > or = ABC light trap with light and CO2 > ABC light trap with light only. A concurrent 5 x 5 test found significant differences in trap catch, where Mosquito Magnet with octenol > New Jersey light trap > or = EPAR Mosquito Killer with CO2 > or = ABC light trap with light and dry ice > Centers for Disease Control (CDC) light trap (manufactured by John W. Hock) with light and octenol. Significant differences in trap catch were noted for several species including: Aedes vexans, Anopheles sinensis, An. yatsushiroensis, An. lesteri, Culex pipiens, and Cx. orientalis. Traps baited with octenol captured significantly fewer Cx. pipiens than those not baited with octenol. Likewise, no Cx. orientalis were captured in octenol-baited traps. Host-seeking activity showed a similar bimodal pattern for all species captured. Results from these field trap evaluations can significantly enhance surveillance efforts. Significantly greater numbers of mosquitoes were captured with mosquito traps using counterflow technology (e.g., Mosquito Magnet and CFG traps) when compared to standard light and carbon dioxide-baited traps. Additionally, field evaluations demonstrate that various traps can be utilized for isolation and detection of arboviruses and other pathogens.

Immunogenicity and protective efficacy of solubilized merozoite-enriched Theileria sergenti immunogens. III. Characterization of immunodominant peptides.

Immunoblot analysis utilizing bovine sera from naturally or experimentally infected with Theileria sergenti were used to determine the immunodominant polypeptides of T. sergenti (Korean isolate). The previously recognized major bands, 18 kDa, 29 kDa, 34 kDa and 45 kDa, were excised after electrophoresis and transfer to PVDF membrane. The individual bands were sequenced. The 34 kDa polypeptide which was the most antigenic and immunogenic peptide was observed in the Western blot. However, Chou-Fasman prediction sites (antigenic site) for antigen determinants of the 45 kDa, 24 kDa, 29 kDa and 18 kDa polypeptide were 6, 4, 2 and 0, respectively. However, the 45 kDa polypeptide showed no reaction with anti-T, sergenti hyperimmune serum.

Evaluation of pH inhibition effect on activated sludge by the pseudo toxic concentration (CPT) concept model.

It is generally accepted that the inhibition effect of pH on activated sludge follows the non-competitive inhibition kinetics. But the non-competitive inhibition kinetic equation cannot be directly applied to pH inhibition, due to the difficulty in quantification of pH as a term of inhibitor concentration. So, many empirical equations were developed especially for acidic condition to describe pH inhibition effect. In this research, the pseudo toxic concentration (CPT) concept model to quantify pH inhibition effect on activated sludge was proposed and compared with other existed models. Prediction of performance, presented by Prediction Accuracy and Prediction Accuracy Index, showed that the CPT concept model can explain the reduction of the maximum specific growth rate (mu max) more accurately than any other models do at a wide range of pH. The CPT concept model was applicable not only to activated sludge but also to specific microorganism groups, such as nitrifier, less acidophilic species and nitrifying biofilm.

Effect of wrist position on the measurement of carpal indices on the lateral radiograph.

The purpose of this study was to find out whether the carpal indices measured on lateral radiographs with a slightly malpositioned wrist are the same as those measured in the true neutral position. Lateral radiographic views of 25 wrists were taken with 5° intervals from 20° of flexion to 20° of extension. Most carpal indices measured in the flexed or extended position were significantly different from the wrist in zero flexion-extension, except scapholunate angle at 5° of extension and scaphocapitate angle at 5° and 10° of flexion. Starting from the flexed position, there was an average of -4.0° change in radioscaphoid angle, -1.0° in scapholunate angle, -1.0° in scaphocapitate angle, +3.0° in radiolunate angle, and +2.0° in lunocapitate angle for each 5° of extension with linear trends. The results from this study suggest that even minimal degrees of flexion-extension can affect the measurements of carpal indices on lateral radiographs.

Differences in post-operative functional disability and patient satisfaction between patients with long (three levels or more) and short (less than three) lumbar fusions.

We examined the differences in post-operative functional disability and patient satisfaction between 56 patients who underwent a lumbar fusion at three or more levels for degenerative disease (group I) and 69 patients, matched by age and gender, who had undergone a one or two level fusion (group II). Their mean age was 66 years (49 to 84) and the mean follow-up was 43 months (24 to 65). The mean pre-operative Oswestry Disability Index (ODI) and visual analogue scale (VAS) for back and leg pain, and the mean post-operative VAS were similar in both groups (p > 0.05), but post-operatively the improvement in ODI was significantly less in group I (40.6%) than in group II (49.5%) (p < 0.001). Of the ten ODI items, patients in group I showed significant problems with lifting, sitting, standing, and travelling (p < 0.05). The most significant differences in the post-operative ODI were observed between patients who had undergone fusion at four or more levels and those who had undergone fusion at less than four levels (p = 0.005). The proportion of patients who were satisfied with their operations was similar in groups I and II (72.7% and 77.0%, respectively) (p = 0.668). The mean number of fused levels was associated with the post-operative ODI (r = 0.266, p = 0.003), but not with the post-operative VAS or satisfaction grade (p > 0.05). Post-operative functional disability was more severe in those with a long-level lumbar fusion, particularly at four or more levels, but patient satisfaction remained similar for those with both long- and short-level fusions.

Purification, cloning, and expression of a pathogen inducible UDP-glucose:Salicylic acid glucosyltransferase from tobacco.

Salicylic acid (SA) plays an important role in plant disease resistance. Inoculation of tobacco leaves with incompatible pathogens triggers the biosynthesis of SA which accumulates primarily as the SA 2-O-beta-D-glucoside (SAG) and glucosyl salicylate (GS). The tobacco UDP-glucose:salicylic acid glucosyltransferase (SA GTase) capable of forming both SAG and GS was purified, characterized, and partially sequenced. It has an apparent molecular mass of 48 kDa, a pH optimum of 7.0, and an isoelectric point at pH 4.4. UDP-glucose was the sole sugar donor for the enzyme. However, SA and several phenolics served as glucose acceptors. The apparent K(m) values for UDP-glucose and SA were 0.27 and 1-2 mM, respectively. Zn(2+) and UDP inhibited its activity. The corresponding cDNA clone which encoded a protein of 459 amino acids was isolated from an SA-induced tobacco cDNA library and overexpressed in Escherichia coli. The recombinant protein catalyzed the formation of SAG and GS, and exhibited a broad specificity to simple phenolics, similar to that of the purified enzyme. Northern blot analysis showed that the SA GTase mRNA was induced both by SA and incompatible pathogens. The rapid induction timing of the mRNA by SA indicates that it belongs to the early SA response genes.

PCR assays for specific and sensitive detection of Pseudomonas tolaasii, the cause of brown blotch disease of mushrooms.

AIMS: The present study describes PCR assays to detect specifically Pseudomonas tolaasii from various samples. METHODS AND RESULTS: Two sets of PCR primers were developed to amplify genes required for tolaasin production. Only a PCR product of 449 bp or 249 bp was produced in PCR reactions with the Pt-1A/Pt-1D1 or Pt-PM/Pt-QM primer sets, respectively, and DNA and cells of Ps. tolaasii. Nested and immunocapture-nested PCR could detect to 3 cells of Ps. tolaasii and amplify the Ps. tolaasii-specific DNA from a sample containing 10 000 times more other bacterial cells than Ps. tolaasii, respectively. CONCLUSIONS: The PCR assays are simple, rapid and reliable methods for detection and identification of Ps. tolaasii. SIGNIFICANCE AND IMPACT OF THE STUDY: The protocols can effectively distinguish Ps. tolaasii from other bacteria and detect Ps. tolaasii from various samples for studying ecology of the bacterium and preventing the use of contaminated water or spawn or medium in mushroom cultivation.