Quorum sensing in Burkholderia cepacia: identification of the LuxRI homologs CepRI

Burkholderia cepacia has emerged as an important pathogen in patients with cystic fibrosis. Many gram-negative pathogens regulate the production of extracellular virulence factors by a cell density-dependent mechanism termed quorum sensing, which involves production of diffusible N-acylated homoserine lactone signal molecules, called autoinducers. Transposon insertion mutants of B. cepacia K56-2 which hyperproduced siderophores on chrome azurol S agar were identified. One mutant, K56-R2, contained an insertion in a luxR homolog that was designated cepR. The flanking DNA region was used to clone the wild-type copy of cepR. Sequence analysis revealed the presence of cepI, a luxI homolog, located 727 bp upstream and divergently transcribed from cepR. A lux box-like sequence was identified upstream of cepI. CepR was 36% identical to Pseudomonas aeruginosa RhlR and 67% identical to SolR of Ralstonia solanacearum. CepI was 38% identical to RhlI and 64% identical to SolI. K56-R2 demonstrated a 67% increase in the production of the siderophore ornibactin, was protease negative on dialyzed brain heart infusion milk agar, and produced 45% less lipase activity in comparison to the parental strain. Complementation of a cepR mutation restored parental levels of ornibactin and protease but not lipase. An N-acylhomoserine lactone was purified from culture fluids and identified as N-octanoylhomoserine lactone. K56-I2, a cepI mutant, was created and shown not to produce N-octanoylhomoserine lactone. K56-I2 hyperproduced ornibactin and did not produce protease. These data suggest both a positive and negative role for cepIR in the regulation of extracellular virulence factor production by B. cepacia.     

Nasal lavage as tool for health effect assessment of photochemical air pollution

In order to achieve esthetically more satisfying results, it has been proposed to place ITI implants with their border between the rough and smooth surfaces below the level of the alveolar crest, thereby obtaining a submucosally located implant shoulder following healing. The aim of the present experimental study was to clinically and radiographically evaluate the tissue response to the placement of one-stage transmucosal implants with the border between the rough and the smooth surfaces sunk by 1 mm into a subcrestal location. 11 patients underwent comprehensive dental care including the placement of 2 implants of the ITI Dental Implant System in the same quadrant (test and control). Randomly assigned control implants were placed according to the manufacturer's instructions, i.e. the border between the rough titanium plasma-sprayed and the smooth polished surfaces precisely at the alveolar crest. At the test implant the apical border of the polished surface was placed approximately 1 mm below the alveolar crest. Probing bone levels were assessed at implant placement (baseline), 4 and 12 months later. Modified plaque and modified gingival indices were recorded at 1, 2, 3, 4 and 12 months. Clinical probing depth and "attachment" levels were measured at 4 and 12 months. All parameters were assessed at 6 sites around each implant. The mean for each implant was calculated and used for analysis. The Wilcoxon matched pairs signed rank test and the Student t-test were applied to detect differences over time and between the test and control implants. At baseline, a mean difference in probing bone level of -0.86 mm (SD 0.43 mm, p < 0.05) was found between test and control implants with the test implants being placed more deeply. Both test and control implants lost a significant amount of clinical bone height during the first 4 months (test 1.16 mm, p < 0.05; control 0.58 mm, p < 0.05). However, only the test implants significantly lost clinical bone height from 4-12 months (test 1.04 mm, p < 0.05; control 0.45 mm, p = 0.08). Overall, the test implants lost 2.26 mm and the control implants 1.02 mm of bone height during the first year of service. On the average, the test implants demonstrated a bone level of 0.38 mm lower than the controls at 12 months. Except for the modified gingival index at 4 months (mean difference 0.21, SD 0.19, p < 0.05), no clinical parameters yielded significant differences between test and control implants at any time. It is concluded that in addition to the crestal bone resorption occurring at implants placed under standard conditions, the bone adjacent to the polished surface of more deeply placed ITI implants is also lost over time. From a biological point of view, the placement of the border between the rough and the smooth surfaces into a subcrestal location should not be recommended.     

Questionnaire study and audit of use of angiotensin converting enzyme inhibitor and monitoring in general practice: the need for guidelines to prevent renal failure

OBJECTIVES: To determine the current pattern of use of angiotensin converting enzyme inhibitor and monitoring of renal function in general practice and to audit all admissions to a regional renal unit for uraemia related to use of these drugs. DESIGN: Postal questionnaire sent to 400 general practitioners; audit of clinical notes of all patients receiving these drugs in one large general practice; audit of all cases of uraemia (creatinine concentration >500 micromol/l) related to treatment presenting to hospital renal services over 12 months. SETTING: General practices in the North Wales health authority and one in central Manchester. Regional renal unit in Salford. MAIN OUTCOME MEASURES: Proportion of general practitioners who regularly monitored renal function before and after initiation of angiotensin converting enzyme inhibitors. Indications for treatment and details of monitoring of renal function in patients receiving these drugs. Incidence of related uraemia and evidence of comorbid disease, other aetiological factors, delayed detection, and patient outcome. RESULTS: 277 (69%) general practitioners replied; 235 (85%) checked renal function before but only 93 (34%) after the start of treatment, and 42 (15%) never checked renal function. Angiotensin converting enzyme inhibitors were prescribed for 162 patients from a total of 3625 aged >35 years (mean age 66.4 (SD 15.9) years). Monitoring of renal function occurred before treatment in 55 (45%) and after start of treatment in 35 (29%) of the 122 patients treated in general practice. Angiotensin converting enzyme inhibitors could be causally implicated in 9 (7%) of 135 admissions for uraemia (mean age 74.2 (7. 2) v 62.1 (2.1) years; P<0.01). 3 patients had renovascular disease and 6 had congestive cardiac failure with another intercurrent illness. Renal function had not been checked in any patient after the start of treatment; mean duration of illness before admission was 10.5 (3.2) days. Mean length of hospital stay was 20.9 (10.4) days; there were 8 survivors. CONCLUSION: Cases of uraemia related to treatment with angiotensin converting enzyme inhibitors are still encountered and are often detected late because of lack of judicious monitoring of renal function in vulnerable, often elderly, patients, especially at times of intercurrent illness. Guidelines for appropriate monitoring of renal function may help to minimise the problem.     

Morphology profiles obtained by reaction‐induced phase separation in epoxy/polysulfone/poly(ether imide) systems

The reaction‐induced phase separation in epoxy/aromatic diamine formulations simultaneously modified with two immiscible thermoplastics (TPs), poly(ether imide) (PEI) and polysulfone (PSF), has been studied. The epoxy monomer was based on the diglycidyl ether of bisphenol A (DGEBA) and the aromatic diamine was 4,4′‐methylenebis(3‐chloro 2,6‐diethylaniline) (MCDEA). Phase‐separation conversions are reported for various PSF/PEI proportions for blends containing 10 wt% total TP. On the basis of phase‐separation results, a conversion–composition phase diagram at 200 °C was compiled. This diagram was used to design particular cure cycles in order to generate different morphologies during the phase‐separation process. It was found that, depending on the PSF/PEI ratio employed, a particulate or a morphology characterized by a distribution of irregular PEI‐rich domains dispersed in an epoxy‐rich phase was obtained for initially miscible blends. Scanning electron microscopy (SEM) characterization revealed that the PEI‐rich phase exhibits a phase‐inverted structure and the epoxy‐rich matrix presents a bimodal size distribution of TP‐rich particles. For PSF/PEI ratios near the miscibility limit, slight temperature change result in morphology profiles. Copyright © 2005 Society of Chemical Industry     

Regulation of S-like ribonuclease levels in Arabidopsis. Antisense inhibition of RNS1 or RNS2 elevates anthocyanin accumulation

The S-like ribonucleases (RNases) RNS1 and RNS2 of Arabidopsis are members of the widespread T2 ribonuclease family, whose members also include the S-RNases, involved in gametophytic self-incompatibility in plants. Both RNS1 and RNS2 mRNAs have been shown previously to be induced by inorganic phosphate (Pi) starvation. In our study we examined this regulation at the protein level and determined the effects of diminishing RNS1 and RNS2 expression using antisense techniques. The Pi-starvation control of RNS1 and RNS2 was confirmed using antibodies specific for each protein. These specific antibodies also demonstrated that RNS1 is secreted, whereas RNS2 is intracellular. By introducing antisense constructs, mRNA accumulation was inhibited by up to 90% for RNS1 and up to 65% for RNS2. These plants contained abnormally high levels of anthocyanins, the production of which is often associated with several forms of stress, including Pi starvation. This effect demonstrates that diminishing the amounts of either RNS1 or RNS2 leads to effects that cannot be compensated for by the actions of other RNases, even though Arabidopsis contains a large number of different RNase activities. These results, together with the differential localization of the proteins, imply that RNS1 and RNS2 have distinct functions in the plant.     

The dermatologist in military operations

The military dermatologist has a specific and significant role in military operations--in time of war as well as in peace. Many dermatologists are unfamiliar with the impact that our specialty and cutaneous disease has upon the ability of the military to fulfill the missions, duties, and responsibilities assigned by our government. This article highlights a few of the recent or ongoing types of military operations in which our specialty plays a prominent part.     

A broad-spectrum microbicide with virucidal activity against sexually transmitted viruses

Sodium dodecyl sulfate (SDS), an alkyl sulfate surfactant derived from an organic alcohol, possesses surfactant properties but also denatures and unfolds both monomeric and subunit proteins. In preliminary experiments, we demonstrated that SDS is a potent inactivator of herpes simplex virus type 2 and human immunodeficiency virus type 1 at concentrations comparable to those used for the surfactant nonoxynol-9. We hypothesized that SDS might be capable of denaturing the capsid proteins of nonenveloped viruses. In this report, we demonstrate inactivation of rabbit, bovine, and human papillomaviruses after brief treatment with dilute solutions of SDS. Effective concentrations were nontoxic to rabbit skin and to split-thickness grafts of human foreskin epithelium. This is the first report of a microbicidal surfactant that will inactivate papillomaviruses. We propose that SDS is now a candidate microbicide for formulation and testing with humans.     

Development of housing programs to aid in the treatment of tuberculosis in homeless individuals: a pilot study

A long forehead disrupts the harmony among the facial components and may contribute to the semblance of facial aging. Slight forehead length disharmony on a senescent face can be corrected by placing the incision at the hairline, elevating the eyebrows through subcutaneous or subgaleal dissection, and removing excess skin without posterior scalp immobilization. For moderate to major reduction of the forehead length, the scalp is elevated back to the occipital region through a pretrichial incision, and relaxation incisions are made at a right angle to the vector of advancement. The entire scalp is then repositioned anteriorly, advancing the hairline caudally and shortening the forehead. Retraction of the scalp or excessive elevation of the eyebrows is prevented by anchoring the galeal fascia to the cranial bone using a bone-tunneling technique in one to three rows. The number of fixation rows is commensurate to the amount of advancement and rigidity of the scalp. The more immobile the scalp preoperatively, the more relaxation incisions and fixation tunnels are necessary. Following caudal repositioning of the scalp, the non-hair-bearing skin is excised, and a meticulous repair is done. These procedures have been performed in 180 patients with a high degree of satisfaction. Temporary hair loss was experienced in one smoker who underwent the most advancement through posterior scalp elevation and continued to smoke postoperatively. Also, on three patients in the subcutaneous forehead rhytidectomy group, two of whom were smokers, delayed healing was observed in the temple area because of compromised circulation requiring secondary revision.     

Ischio-vertebral dysplasia: a distinct entity

A total of 20 isolates of Blastocystis were characterized using a single set of polymerase chain reaction (PCR) primers. The amplification product revealed five types of pattern. All four isolates from Singapore yielded PCR products quite different from those of the local isolates. However, most of the local isolates showed a major product at either 280 or 500 bp, or both. We also suspected that the amplification product detected at 280 bp might be an indicator of the pathogenicity of this parasite. One isolate (M12) obtained from a monkey showed patterns similar to those of human isolates (10203 and KP1) and probably belongs to the same strain. The results indicate that the intraspecific or interstrain variations in these 20 Blastocystis isolates belong to 5 different patterns. The differences among isolates of the same strain revealed by the presence or absence of certain amplification products showed further intrastrain variations in this parasite.