Identification and characterization of a two-component regulatory system involved in invasion of eukaryotic cells and heavy-metal resistance in Burkholderia pseudomallei

Burkholderia pseudomallei is the causative agent of melioidosis, a disease increasingly recognized as an important cause of morbidity and mortality in many regions of the world. B. pseudomallei is a facultative intracellular pathogen capable of invading eukaryotic cells. We used Tn5-OT182 mutagenesis to generate mutants deficient in the ability to invade a human type II pneumocyte cell line (A549 cells). One of these mutants, AJ1D8, exhibited approximately 10% of the ability of the parental strain, 1026b, to invade A549 cells. There was no difference in the abilities of 1026b and AJ1D8 to resist killing by RAW macrophages or the human defensin HNP-1. The nucleotide sequence flanking the Tn5-OT182 integration in AJ1D8 was determined, and two open reading frames were identified. The predicted proteins shared considerable homology with two-component regulatory systems involved in the regulation of heavy-metal resistance in other organisms. AJ1D8 was 16-fold more sensitive to Cd2+ and twofold more sensitive to Zn2+ than was 1026b but was not sensitive to any of the other heavy metals examined. The B. pseudomallei two-component regulatory system, termed irlRS, complemented the invasion-deficient and heavy-metal-sensitive phenotype of AJ1D8 in trans. There was no significant difference between the virulence of AJ1D8 and that of 1026b in infant diabetic rats and Syrian hamsters, suggesting that the irlRS locus is probably not a virulence determinant in these animal models of acute B. pseudomallei infection.     

The renal and hepatic distribution of Bence Jones proteins depends on glycosylation: a scintigraphic study in rats

The aim of the present study was to evaluate renal and liver distribution of two monoclonal immunoglobulin light chains. The chains were purified individually from the urine of patients with multiple myeloma and characterized as lambda light chains with a molecular mass of 28 kDa. They were named BJg (high amount of galactose residues exposed) and BJs (sialic acid residues exposed) on the basis of carbohydrate content. A scintigraphic study was performed on male Wistar rats weighing 250 g for 60 min after i.v. administration of 1 mg of each protein (7.4 MBq), as the intact proteins and also after carbohydrate oxidation. Images were obtained with a Siemens gamma camera with a high-resolution collimator and processed with a MicroDelta system. Hepatic and renal distribution were established and are reported as percent of injected dose. Liver uptake of BJg was significantly higher than liver uptake of BJs (94.3 vs 81.4%) (P < 0.05). This contributed to its greater removal from the intravascular compartment, and consequently lower kidney accumulation of BJg in comparison to BJs (5.7 vs 18.6%) (P < 0.05). After carbohydrate oxidation, there was a decrease in hepatic accumulation of both proteins and consequently a higher renal overload. The tissue distribution of periodate-treated BJg was similar to that of native BJs: 82.7 vs 81.4% in the liver and 17.3 vs 18.6% in the kidneys. These observations indicate the important role of sugar residues of Bence Jones proteins for their recognition by specific membrane receptors, which leads to differential tissue accumulation and possible toxicity.     

Psychosocial rehabilitation of cranial trauma and stroke patients

This paper presents the results of a longitudinal psychosocial study of 22 cranial trauma patients and 14 stroke patients from the time preceding injury (using retrospective data), through a 4-5 month intensive rehabilitation programme, to a follow-up 1 year after completion of the programme. Although the two groups of patients differed on several demographic and medical characteristics, essentially similar patterns for psychosocial decline following injury and improvement following rehabilitation could be observed. For both groups, the proportion in marital or cohabitational relationships returned to pre-injury levels, and for both groups the proportion requiring assistance in their living situation declined following rehabilitation, as did use of the health services. Virtually all patients in both groups had been in employment or undergoing education at the time of the injury, and although this percentage declined in practice to a small minority of both groups post-injury, there was a significant increase in the proportions working or in education following the rehabilitation programme. Similarly, the pattern of leisure-time activities in both groups declined post-injury and was restored following rehabilitation. Since both groups entered the programme at over 2.5 years post-injury, these generally encouraging results seem less likely to reflect spontaneous recovery than a beneficial effect of the programme itself.     

Mapping the chicken genome: problems and perspectives

Various molecular methods are now used to map the chicken genome, including chromosome scraping, flow cytofluorimetry, zonal centrifugation, construction of chromosome-specific libraries, genetic analysis with polymorphic DNA markers, and in situ hybridization. Two main drawbacks are characteristic of existing maps of chicken chromosomes. First, classic genetic maps (i.e., linkage groups of genes for morphological, physiological, and biochemical characters), physical maps of chromosomes, and new genetic maps constructed on the basis of polymorphic DNA markers (RFLP, RAPD, VNTR, SSR, and CR1-PCR) do not coordinate with one another. Second, a relatively low number of genes is present in classic genetic maps and physical chromosome maps. Application of cytogenetic methods to chromosome mapping in birds is limited because of some specific features characteristic of the organization of avian genomes. For the same reason, studying the location and expression of avian genes is very important. Since mammalian and avian genomes differ in structure, revealing their possible common functional characteristics will provide for a better understanding of the general mechanisms that control biologically important characters in higher animals.     

Noninvasive determination of cardiac output in patients with severe airflow limitation

The noninvasive measurement of cardiac output (Q) by the Indirect Fick CO2-rebreathing technique requires mixed venous P CO2 (P CO2) to be determined by the rebreathing maneuver, and Pa CO2 to be estimated from end-tidal P CO2 (PET CO2). Previous work has suggested that although P CO2 can be determined, Pa CO2 cannot be accurately estimated in patients with significant airflow limitation. Nineteen patients with cystic fibrosis who had severe airflow limitation (%FEV1, 29.3 +/- 7.12 SD) were studied during steady-state exercise at 50% of their measured maximal work capacity. Estimated Pa CO2 was slightly lower than Pa CO2 measured from blood samples obtained from an indwelling arterial catheter (measured: 45.2 +/- 4.92; estimate: 42.7 +/- 5.68 mm Hg). To calculate arterial blood content, the values derived from Pa CO2, pH, hemoglobin (Hb), and O2 saturation were compared with those derived from PET CO2 and O2 saturation, where (1) pH was assumed to be 7.40 and Hb was measured, and (2) pH was assumed to be 7.40 and Hb was assumed to be 15 g/dl (measured mean pH, 7.34; Hb, 14.4 g/dl). No difference in arterial CO2 content was seen between the three methods (measured: 47.53 +/- 5.17; estimate 1: 49.57 +/- 6.58; estimate 2: 49.12 +/- 6.61 ml/100 ml). As pH and Hb can also affect mixed venous CO2 content, the effect on Q was also assessed. Both estimates fit closely with measured Q (r2=0.77 and 0.76), with intercepts not different from zero and slopes not different from 1, and coefficients of variation of 13.5 and 14.6%. When viewed with regard to the confidence intervals for Q as a function of O2 consumption, Q was altered to a minor extent. We conclude that the use of PET CO2 to estimate Pa CO2 can give reasonable values for Q determined noninvasively in patients with severe airflow limitation.     

A comparative study of color measurement lntstrumentation

A multiplant Quality Improvement Team [QIT] was firmed to develop and implement an evaluation program for various color measurement .systems as potential replacements for the then-current aging systems. The emphasis qf this article is the analytical methodology utilized to evaluate the various color systems. The evaluation program consisted cf two phases. Phase I was a general overview/review of several systems, while Phase II was an extensive internal comparative evaluation measurement systems. These were Milton-Roy's ColorMate HDS, HunterLab's Ultrascan, Datacolor's CS-5, and BYK-Gardner's The Color Sphere [TCS]. The main comparison criteria were interinstrument agreement [agreement between two instruments ofthe same system], user-friendly software and computer interface capability, vendor amenability to a long-term logistical and maintenance relationship, and price. All systems were evaluated by duplicate measurements on various color tiles, yarns, and polymer flakes-over 1600 measurements on each system. The systems were compared with an instrument matrix, a decision matrix, and a product matrix. The instrument matrix was a comparison qfinstrument parameters, software/math treatments, and economics. The decision matrix was a forced ranking of each system by each criteria category [1–4 scale, with 1 representing the best and 4 representing the worst]. The product matrix accentuated the relative importance ofone criterion category over another by multiplying the forced ranking by the criticality of the category. The criticality of a given category wus determined by consensus within the QIT. Thr combination qf the three matrices allowed the evaluator[.s] t o select the color rneasuremmt system that best satixfied the color measurement needs and requirements of their facility and their products. For this evaluation, all ofthe evaluated systems were superior to the then-current agingsystems. As a result of this methodology, one instrument emerged as clearly superior. © 1994 John Wrley & Sons, Inc.     

A novel model to study the effects of burn lymph on pulmonary vascular hemodynamic variables

The possible hypocholesterolaemic properties of milk and fermented milk products have been investigated in groups of albino rats given a basal diet, basal diet plus cholesterol, and basal diet plus cholesterol together with whole milk or standard or bifidus yogurt. The yogurts were fortified with skim milk powder, condensed whey or lactose-hydrolysed condensed whey. After 30 d, triacylglycerols, total cholesterol, HDL-cholesterol and LDL-cholesterol were measured in serum. Whole milk and ordinary yogurt had no hypocholesterolaemic effect, but standard yogurt containing lactose-hydrolysed condensed whey and all bifidus yogurts lowered serum cholesterol. In general, yogurts changed HDL-cholesterol little, but tended to raise triacylglycerols. There was marked lowering of LDL-cholesterol in rats given either type of yogurt fortified with whey proteins. This study has demonstrated in a rat model that bifidus yogurts and yogurts fortified with whey proteins can reduce total and LDL-cholesterol, and suggests that if they have the same effect in human subjects they have potential value in cholesterol-lowering diets.