Diagnosing infections--current and anticipated technologies for point-of-care diagnostics and home-based testing.

In recent years, we have witnessed many transitions in healthcare systems around the globe. For example, population expansion and ageing, and the human immunodeficiency virus (HIV)-AIDS epidemics, have exerted pressure to decentralize the practice of healthcare outside of traditional settings to bring care to those in need. Upstream of patient management, diagnosis is aimed at adequately orienting medical decisions, and considerable efforts have been made to make this process faster and more efficient. However, there are several diseases and medical conditions that may/will benefit from technologies and tests that can be performed closer to the patient, at the point of care or even in the home. In this review, and in light of the paradox that technology and assay developers and healthcare officials must take into consideration for advancing human health in developed and developing countries, we present an overview of rapid diagnosis of infectious diseases at the point of care and of technologies that may contribute to enhancement of the worldwide point-of-care testing market.

Assessment of microbiological quality of drinking water from household tanks in Bermuda.

Bermuda residents collect rainwater from rooftops to fulfil their freshwater needs. The objective of this study was to assess the microbiological quality of drinking water in household tanks throughout Bermuda. The tanks surveyed were selected randomly from the electoral register. Governmental officers visited the selected household (n = 102) to collect water samples and administer a short questionnaire about the tank characteristics, the residents' habits in terms of water use, and general information on the water collecting system and its maintenance. At the same time, water samples were collected for analysis and total coliforms and Escherichia coli were determined by 2 methods (membrane filtration and culture on chromogenic media, Colilert kit). Results from the 2 methods were highly correlated and showed that approximately 90% of the samples analysed were contaminated with total coliforms in concentrations exceeding 10 CFU/100 mL, and approximately 66% of samples showed contamination with E. coli. Tank cleaning in the year prior to sampling seems to protect against water contamination. If rainwater collection from roofs is the most efficient mean for providing freshwater to Bermudians, it must not be considered a source of high quality drinking water because of the high levels of microbial contamination.

A search for specific and common susceptibility loci for schizophrenia and bipolar disorder: a linkage study in 13 target chromosomes.

We report the first stage of a genome scan of schizophrenia (SZ) and bipolar disorder (BP) covering 18 candidate chromosomal areas. In addition to testing susceptibility loci that are specific to each disorder, we tested the hypothesis that some susceptibility loci might be common to both disorders. A total of 480 individuals from 21 multigenerational pedigrees of Eastern Québec were evaluated by means of a consensus best-estimate diagnosis made blind to diagnoses in relatives and were genotyped with 220 microsatellite markers. Two-point and multipoint model-based linkage analyses were performed and mod scores (Z, for max Z(max)) are reported. The strongest linkage signals were detected at D18S1145 (in 18q12; Z = 4.03) for BP, and at D6S334 (in 6p 22-24; Z(het) = 3.47; alpha = 0.66) for SZ. Three other chromosomal areas (3q, 10p, and 21q) yielded linkage signals. Chromosomes 3p, 4p, 5p, 5q, 6q, 8p, 9q, 11q, 11p, 12q, 13q, 18p and 22q showed no evidence of linkage. The 18q12 results met the Lander and Kruglyak (1995) criterion for a genome-wide significant linkage and suggested that this susceptibility region may be shared by SZ and BP. The 6p finding provided confirmatory evidence of linkage for SZ. Our results suggest that both specific and common susceptibility loci must be searched for SZ and BP.

The contribution of arginine residues within the P6-P1 region of alpha 1-antitrypsin to its reaction with furin.

A series of mutants incorporating furin recognition sequences within the P6-P1 region of the reactive site loop of alpha(1)-antitrypsin were constructed. Variants containing different combinations of basic residues in the P1, P2, P4, and P6 positions replacing the wild type (P6)LEAIPM(P1) sequence were evaluated for their capacity to establish SDS-resistant complexes with furin, to affect association rate constants (k(ass) and k'(ass)), or to inhibit furin-dependent proteolysis of a model precursor in vivo. Each variant abolished processing of pro-von Willebrand factor in transfected hEK293 cells. The k(ass) of all variants were found to be similar (1.1-1.7 x 10(6) m(-1) s(-1)) except for one mutant, RERIRR, which had a k(ass) of 3.3 x 10(5) m(-1) s(-1). However, the stoichiometry of inhibition varied with values ranging from 2.9 to >24, indicating rapid formation of the acyl-enzyme intermediate (high k'(ass)). Moreover, those variants having high stoichiometry of inhibition values were accompanied by the rapid formation of cleaved forms of the inhibitors. The data suggest that the rate of conversion of the acyl-enzyme (EI') into the highly stable complex (EI*) was affected by replacement of specific residues within the reactive site loop. Taken together, the results reveal how furin recognition sequences within the context of the biochemical properties of serpins will play a role in the capacity of the protein to follow either the inhibitory or the substrate pathway.

Strong dependence of rain-induced lidar depolarization on the illumination angle: experimental evidence and geometrical-optics interpretation.

Backscatter and depolarization lidar measurements from clouds and precipitation are reported as functions of the elevation angle of the pointing lidar direction. We recorded the data by scanning the lidar beam (Nd:YAG) at a constant angular speed of ~3.5 degrees /s while operating at a repetition rate of 10 Hz. We show that in rain there is an evident and at times spectacular dependence on the elevation angle. That dependence appears to be sensitive to raindrop size. We have developed a three-dimensional polarization-dependent ray-tracing algorithm to calculate the backscatter and the depolarization ratio by large nonspherical droplets. We have applied it to raindrop shapes derived from existing static and dynamic (oscillating) models. We show that many of the observed complex backscatter and depolarization features can be interpreted to a good extent by geometrical optics. These results suggest that there is a definite need for more extensive calculations of the scattering phase matrix elements for large deformed raindrops as functions of the direction of illumination. Obvious applications are retrieval of information on the liquid-solid phase of precipitation and on the size and the vibration state of raindrops.

Significant linkage for Tourette syndrome in a large French Canadian family.

Family and twin studies provide strong evidence that genetic factors are involved in the transmission of Gilles de la Tourette syndrome (TS) and related psychiatric disorders. To detect the underlying susceptibility gene(s) for TS, we performed linkage analysis in one large French Canadian family (127 members) from the Charlevoix region, in which 20 family members were definitely affected by TS and 20 others showed related tic disorders. Using model-based linkage analysis, we observed a LOD score of 3.24 on chromosome 11 (11q23). This result was obtained in a multipoint approach involving marker D11S1377, the marker for which significant linkage disequilibrium with TS recently has been detected in an Afrikaner population. Altogether, 25 markers were studied, and, for level of significance, we derived a criterion that took into account the multiple testing arising from the use of three phenotype definitions and three modes of inheritance, a procedure that yielded a LOD score of 3.18. Hence, even after adjustment for multiple testing, the present study shows statistically significant evidence for genetic linkage with TS.

Retrieval of droplet-size density distribution from multiple-field-of-view cross-polarized lidar signals: theory and experimental validation.

Multiple-field-of-view (MFOV) secondary-polarization lidar signals are used to calculate the particle-size density distribution (PSD) at the base of a cloud. At the cloud base, multiple scattering is weak and single backscattering is predominant by many orders of magnitude. Because secondary polarization is a direct measure of multiple scattering, it is therefore advantageous to use secondary polarization. A mathematical relation among the PSD, the lidar fields of view, the scattering angles, and the angular depolarization is derived to facilitate use of secondary polarization. The model is supported by experimental MFOV lidar measurements carried out in a controlled environment, and its limitations and restrictions are discussed.

Multiple-scattering effect on ozone retrieval from space-based differential absorption lidar measurements.

Single-scattering and multiple-scattering lidar signals are calculated for a spaceborne differential absorption lidar system for global ozone measurements at the on and off wavelength pair at 305 and 315 nm. The effect of multiple scattering is found to be negligible on stratospheric and tropospheric ozone retrieval under background stratospheric aerosol. Under low-visibility conditions in the planetary boundary layer the presence of multiple scattering causes an overestimation in maritime aerosol and an underestimation in urban as well as in rural aerosol. This effect is also examined in three cirrus models. The multiple scattering does not permit accurate ozone retrieval within cirrus; however, below it the solution recovers somewhat with generally an underestimation depending on the type and density of cirrus. The effect of aerosol and Rayleigh extinction on the ozone retrieval is also discussed.

6p24-22 region and major psychoses in the Eastern Quebec population. Le Groupe IREP.

Recent reports of a linkage trend in 6p24-22 for schizophrenia (SZ), in different samples, were tempered by the concurrent evidence of negative reports in other samples. In the studies showing positive results, different definitions of affection and a wide spectrum of diagnoses were used. Our objectives were not only to test for linkage at 6p24-22 in the Eastern Quebec population, but also to test whether this putative vulnerability locus was either selectively linked to schizophrenia (SZ), or to bipolar disorder (BP), or to both major psychoses. Parametric and nonparametric linkage analyses with 12 microsatellite markers in 6p24-p22 were performed on a sample of 18 large multigenerational pedigrees (N = 354) either affected by SZ, or by BP, or equally affected by both major psychoses (i.e., mixed pedigrees). Three affection definitions were usually tested in our program: one on schizophrenia (SZ), one on bipolar disorder (BP), and one that comprised SZ and BP under the hypothesis of a susceptibility locus common to both in major psychoses (common locus, CL). The results of parametric analyses did not support a major gene hypothesis. However, in one large mixed pedigree (#151), we observed with the common locus phenotype (CL) lod scores of 2.49 and 2.15, respectively, at the D6S296 and D6S277 loci under a dominant model. Our data suggest the presence of a potential vulnerability locus at 6p24-22 that could be related to both schizophrenia and bipolar affective disorder. These results may be seen as congruent with former studies that used schizoaffective as well as schizophrenia diagnoses as entry criteria for the affected families, and used an affection definition that comprised affective psychoses as well as schizophrenia.

Lidar effective multiple-scattering coefficients in cirrus clouds.

We delimit a regime, valid for most ground-based lidar probings of cirrus clouds, in which the field-of-view dependence of multiple scattering reaches a plateau. In this regime and assuming the phase function to be constant around pi, we formally demonstrate Platt's modification of the single-scattering lidar equation, with a parameter eta(P) accounting for the reduction of the effective scattering coefficient defined so that (1 - eta(P)) is the amount of energy scattered in the forward peak. Then, to cope with nonconstant backscattering functions, we discuss the introduction of an effective backscattering coefficient that is an average of the scattering probabilities around pi.

Childhood/early adolescence-onset and adult-onset schizophrenia. Heterogeneity at the dopamine D3 receptor gene.

BACKGROUND: The dopamine D3 receptor gene (DRD3) is a meaningful candidate gene because it unifies the dopamine and the limbic hypotheses for schizophrenia. We tested for an allelic association between schizophrenia and the DRD3 Mscl alleles, hypothesising heterogeneity between childhood/early adolescence-onset schizophrenia (CO-SZ) and adult-onset schizophrenia (A-SZ). METHOD: The frequencies of the DRD3 alleles were compared between 70 DSM-III-R schizophrenics (35 CO-SZ and 35 A-SZ) and 79 controls. RESULTS: Compared with the controls, the subsample of A-SZ, but not CO-SZ, showed an over-proportion (P = 0.025) of allele 1. The association was not found in the total sample, combining the two subsamples. CONCLUSIONS: Consistently with former studies, our data suggest an aetiological heterogeneity between CO-SZ and A-SZ and a possible specificity of the excess of allele 1 to the familial form of schizophrenia and to schizophrenia with a better outcome.

Negative outcomes: what is the evidence on self-administered treatments?

D.C. Mohr (1995) suggested that high deterioration rates may occur in self-directed treatments. The investigators examined data from 5 studies of self-administered treatment for depression and found in contrast much lower rates (9% vs. 19%) than those cited in Mohr's review. The negative response rates for the self-administered treatments compared favorably with the negative response rates in the therapist-administered treatments provided in the National Institute of Mental Health Treatment of Depression Collaborative Research Program. The findings indicate that it may be the manner in which participants are prepared for self-administered treatment that is critical.

Multiple-scattering lidar equation.

A multiple-scattering lidar equation is derived from a phenomenological representation of the scattering processes. The contributions are separated into the unscattered, singly scattered, and multiply scattered illumination of the scattering volume, a single backscattering reflection from that volume, and the unscattered and multiply scattered propagation back to the receiver. The equation is obtained in the form of analytic expressions that explicitly show the signal dependence on the extinction coefficient, the effective particle size, the range, and the receiver field of view. Consistent agreement is found with Monte Carlo calculations and published laboratory measurements. Numerical simulations demonstrate that measurements made at three or more fields of view can be inverted to solve for the extinction coefficient and the effective particle radius. The multiple scatterings taken into account in the proposed equation are the small-angle diffraction scatterings; the wide-angle scatterings caused by refraction and reflection are considered lost, except for one backscattering at an angle close to 180°. Consequently, the equation is applicable to cases in which the projection of the lidar receiver field of view on the cloud is of the order of the angular width of the diffraction peak of the phase function times the penetration depth into the cloud.

Multiply scattered aerosol lidar returns: inversion method and comparison with in situ measurements.

A novel aerosol lidar inversion method based on the use of multiple-scattering contributions measured by a multiple-field-of-view receiver is proposed. The method requires assumptions that restrict applications to aerosol particles large enough to give rise to measurable multiple scattering and depends on parameters that must be specified empirically but that have an uncertainty range of much less than the boundary value and the backscatter-to-extinction ratio of the conventional single-scattering inversion methods. The proposed method is applied to cloud measurements. The solutions obtained are the profiles of the scattering coefficient and the effective diameter of the cloud droplets. With mild assumptions on the form of the function, the full-size distribution is estimated at each range position from which the extinction coefficient at any visible and infrared wavelength and the liquid water content can be determined. Typical results on slant-path-integrated optical depth, vertical extinction profiles, and fluctuation statistics are compared with in situ data obtained in two field experiments. The inversion works well in all cases reported here, i.e., for water clouds at optical depths between ~0.1 and ~4.

Multiple field of view lidar returns from atmospheric aerosols.

As a laser pulse propagates into the atmosphere, it becomes broader in the lateral direction as a result of scattering by aerosols. The laser pulse may be described as the superposition of a central, unscattered component of reduced intensity and a surrounding scattered component. A multiple field of view lidar has been developed that makes simultaneous measurements of the backscattered power from the central pulse and multiply scattered power arising from the scattered component. Measurements from various types of atmospheric aerosols and precipitation are presented and compared with simulated returns. The results show how the multiply scattered signals are influenced by the distribution of the aerosols along the lidar path, the characteristic size of the aerosols, and the optical depth. It is shown that the multiple field of view lidar can provide meaningful, additional information about the aerosols that is not available from a conventional single field of view lidar.

Characterization of In0 of Pseudomonas aeruginosa plasmid pVS1, an ancestor of integrons of multiresistance plasmids and transposons of gram-negative bacteria.

Many multiresistance plasmids and transposons of gram-negative bacteria carry related DNA elements that appear to have evolved from a common ancestor by site-specific integration of discrete cassettes containing antibiotic resistance genes or sequences of unknown function. The site of integration is flanked by conserved segments coding for an integraselike protein and for sulfonamide resistance, respectively. These segments, together with the antibiotic resistance genes between them, have been termed integrons (H. W. Stokes and R. M. Hall, Mol. Microbiol. 3:1669-1683, 1989). We report here the characterization of an integron, In0, from Pseudomonas aeruginosa plasmid pVS1, which has an unoccupied integration site and hence may be an ancestor of more complex integrons. Codon usage of the integrase (int) and sulfonamide resistance (sul1) genes carried by this integron suggests a common origin. This contrasts with the codon usage of other antibiotic resistance genes that were presumably integrated later as cassettes during the evolution and spread of these DNA elements. We propose evolutionary schemes for (i) the genesis of the integrons by the site-specific integration of antibiotic resistance genes and (ii) the evolution of the integrons of multiresistance plasmids and transposons, in relation to the evolution of transposons related to Tn21.

Extinction of visible and infrared beams by falling snow.

Classical optics holds that the extinction cross of particles should be equal to twice their geometric cross section, in the limit where the particles are much larger than the wavelength. It follows that the extinction coefficient of such large scatterers should be independent of wavelength. Snowflakes are much larger than the wavelengths of visible and infrared radiation, yet many investigators have found that the visible and infrared extinction coefficient of falling snow measured with transmissometers is wavelength dependent. This dependency is known to be a result of the scattering contribution to the transmissometer signal. Furthermore, many measurements in the visible and infrared show that extinction values measured simultaneously with two transmissometers are linearly related up to at least 12 km(-1). The slope depends on the wavelengths and optical characteristics of the transmissometers. We show that for small values of extinction, the observations can be explained by taking into account single-scattering contributions to transmissometer signals. For high values of extinction, a multiplescattering model gives good agreement with measurements.

Characterization of the nonenzymatic chloramphenicol resistance (cmlA) gene of the In4 integron of Tn1696: similarity of the product to transmembrane transport proteins.

Integrons constitute a novel family of DNA elements which evolved by site-specific integration of discrete units between two conserved segments. On the In4 integron of Tn1696, a precisely inserted gene cassette of 1,549 bp conferring nonenzymatic chloramphenicol resistance (cmlA) is present between the streptomycin-spectinomycin resistance (aadA2) gene cassette and the 3'-conserved segment of the integron. In this study, we present the nucleotide sequence of the cmlA gene cassette of Tn1696, show its similarity to bacterial efflux systems and other transport proteins, and present evidence for alterations that its expression exerts on bacterial membranes. The cmlA gene cassette apparently carries its own promoter(s), a situation that has not heretofore been observed in the integrons of multiresistance plasmids and transposons of gram-negative bacteria. One or more of these promoters were shown to be functionally active in expressing a cat marker gene from promoter-probe vectors. The putative CmlA polypeptide appears to provoke a reduction of the content of the major porins OmpA and OmpC.

Characterization and expression of the prostatic arginine esterase gene, a canine glandular kallikrein.

The prostatic arginine esterase gene was isolated from a genomic library prepared with dog liver DNA in lambda EMBL3. The selected clone contained an insert of approximately 17 kb which included the whole coding portion of arginine esterase mRNA (5 exons plus 4 introns), 2 kb upstream from the initiation site and 12 kb downstream from the polyadenylation site. The intron-exon boundaries were identical to all known mammalian kallikrein genes. The deduced amino acid sequence indicated a high degree of identity (51-61%) with other kallikreins expressed not only in the prostate but also in the pancreas of various animal species. The 5'-flanking sequences contained potential regulatory elements such as a variant TATA box (TTTAAA), a CCAAT box, a SP1 transcriptional factor binding site (GGGCGG), and two TGTCCT motifs resembling glucocorticoid response elements. Southern blot analysis with an amplified cDNA fragment of 487 bp corresponding to the 5' portion of the mRNA and with a DNA probe from a different portion of the arginine esterase gene indicated the presence of two to three homologous genes in the canine genome while in a previous study a single band was detected using a 400-bp arginine esterase cDNA corresponding to the 3' portion of the mRNA. These results suggest that the arginine esterase gene belongs to a small kallikrein gene family. Arginine esterase mRNA is expressed primarily in the prostate but also at an extremely low level (approximately a thousandfold less) in several other tissues including the liver, the gracilis thigh muscle, the kidney, and the pancreas.