Drivers under the influence of drugs of abuse: quantification of cocaine and impaired driving.

In recent years, the interest in oral fluid as a biological matrix has significantly increased, particularly for detecting driving under the influence of drugs. In this study, the concentration of cocaine and its relationship with clinical symptoms in drivers suspected of driving under the influence of drugs was evaluated. A total of 154 samples of oral fluid, which tested positive for cocaine in previous immunoassay screening, Cozart Drug Detector System, were confirmed using gas chromatography/mass spectrometry method. In Catalonia, during 2007-2010, there were 1791 samples positive for cocaine among a total of 3468 samples taken from drivers who tested positive for any drug of abuse. The evaluation of clinical symptoms was through a questionnaire that was filled in by the police officers who collected the samples. The mean concentration of cocaine was 4.11 mg/l and median concentration was 0.38 mg/l (range 0.01-345.64 mg/l). Clinical impairment symptoms such as motor coordination, walking, speech, mood and state of pupils were not significant. The testing of oral fluids presents fewer ethical problems than blood or urine.

Role of medical history and medication use in the aetiology of upper aerodigestive tract cancers in Europe: the ARCAGE study.

BACKGROUND: The study aimed to investigate the role of medical history (skin warts, Candida albicans, herpetic lesions, heartburn, regurgitation) and medication use (for heartburn; for regurgitation; aspirin) in the aetiology of upper aerodigestive tract (UADT) cancer. METHODS: A multicentre (10 European countries) case-control study [Alcohol-Related CAncers and GEnetic susceptibility (ARCAGE) project]. RESULTS: There were 1779 cases of UADT cancer and 1993 controls. History of warts or C. albicans infection was associated with a reduced risk [odds ratio (OR) 0.80, 95% confidence interval (CI) 0.68-0.94 and OR 0.73, 95% CI 0.60-0.89, respectively] but there was no association with herpetic lesions, heartburn, regurgitation or medication for related symptoms. Regurgitation was associated with an increased risk for cancer of the oesophagus (OR 1.47, 95% CI 0.98-2.21). Regular aspirin use was not associated with risk of UADT cancer overall but was associated with a reduced risk for cancer of oesophagus (OR 0.51, 95% CI 0.28-0.96), hypopharynx (OR 0.53, 95% CI 0.28-1.02) and larynx (OR 0.74, 95% CI 0.54-1.01). CONCLUSIONS: A history of some infections appears to be a marker for decreased risk of UADT cancer. The role of medical history and medication use varied by UADT subsites with aspirin use associated with a decreased risk of oesophageal cancer and suggestive of a decreased risk of hypopharyngeal and laryngeal cancers.

Life course social mobility and risk of upper aerodigestive tract cancer in men.

The aim of this study was to explore associations between social mobility and tumours of the upper aero-digestive tract (UADT), focussing on life-course transitions in social prestige (SP) based on occupational history. 1,796 cases diagnosed between 1993 and 2005 in ten European countries were compared with 1585 controls. SP was classified by the Standard International Occupational Prestige Scale (SIOPS) based on job histories. SIOPS was categorised in high (H), medium (M) and low (L). Time weighted average achieved and transitions between SP with nine trajectories: H --> H, H --> M, H --> L, M --> H, M --> M, M --> L, L --> H, L --> M and L --> L were analysed. Odds ratios (ORs) and 95%-confidence intervals [95%-CIs] were estimated with logistic regression models including age, consumption of fruits/vegetables, study centre, smoking and alcohol consumption. The adjusted OR for the lowest versus the highest of three categories (time weighted average of SP) was 1.28 [1.04-1.56]. The distance of SP widened between cases and controls during working life. The downward trajectory H --> L gave an OR of 1.71 [0.75-3.87] as compared to H --> H. Subjects with M --> M and L --> L trajectories ORs were also elevated relative to subjects with H --> H trajectories. The association between SP and UADT is not fully explained by confounding factors. Downward social trajectory during the life course may be an independent risk factor for UADT cancers.

Genetic susceptibility factors in type 1 diabetes: linkage, disequilibrium and functional analyses.

Continuing progress has been made in elucidating the genetic factors involved in type 1 diabetes (insulin-dependent diabetes mellitus [IDDM]) in the past year. Two genome scans suggested additional susceptibility intervals and provided supporting evidence for several previously reported linkages. Other studies focused on the confirmation of linkage using multipoint sibpair analyses with densely spaced markers and multiethnic collections of families. Although significant and consistent linkage evidence was reported for the susceptibility intervals IDDM8 (on human chromosome 6q27), IDDM4 (on 11q) and IDDM5 (on 6q25), evidence for most other intervals varies in different data sets -probably due to a weak effect of the disease genes, genetic heterogeneity or random variation. Linkage disequilibrium mapping has become an increasingly important tool for both the confirmation and fine-mapping of susceptibility intervals, as well as identification of etiological mutations. Functional studies indicate, firstly, that the susceptible and protective HLA class II molecules HLA-DR and -DQ bind and present nonoverlapping peptides and, secondly, that the variable number of tandem repeats at the 5' end of the insulin gene (susceptibility interval IDDM2) regulates insulin expression in the thymus.

Agreement Between Computed Tomography and Pathologic Nodule Counts in Colorectal Lung Metastases.

BACKGROUND: Computed tomography is the most common technique used to estimate the number of pulmonary metastases and their resectability. A lack of agreement between radiologic and surgical pathologic findings could potentially lead to incomplete resection or to rejection of patients for potentially curative treatments. The objective of this study was to estimate the disagreement between the number of radiologic lesions and the number of histologically confirmed malignant lesions excised from patients with pulmonary metastases from colorectal cancer. METHODS: This was a multicenter longitudinal study using a national registry. All patients underwent open surgery for pulmonary metastasectomy. RESULTS: Radiologic unilateral involvement was documented in 345 of 404 patients (85%); 253 (73%) presented with single nodules. The radiologic and malignant pathologic findings were concordant in 316 (78%) patients. The two independent predictors of discordance between computed tomography and the number of pathologic metastases were the bilateral involvement and the number of radiologic nodules. This model explained 28% of the variability in the disagreement frequency and discriminated between agreement and disagreement in 85% of the patients. Discrepancies increased with the nodule count with an odds ratio of 6.17 (95% confidence interval, 4.08 to 9.33) per additional nodule. For similar nodule counts, a lower disagreement frequency was observed among bilateral cases (odds ratio, 0.2; 95% confidence interval, 0.07 to 0.55). CONCLUSIONS: Differences between the radiologic and pathologic findings were documented in 1 of every 5 patients. The correlation was very accurate in patients with single radiologic nodules. However, half of the patients with more nodules showed discrepancies.

Altered protein synthesis in a cell-free system exposed to a sinusoidal magnetic field.

This report describes a new approach for examining weak extremely low frequency (ELF) electric and magnetic field interactions with living systems that exploits a cell-free transcription/translation system derived from Escherichia coli. Using two-dimensional polyacrylamide gel electrophoresis we previously had determined that the level of the alpha subunit of RNA polymerase in intact E. coli was elevated by exposure to weak ELF magnetic fields. In this paper, plasmids containing the alpha, or both the beta,beta' subunits of the RNA polymerase from E. coli were placed into a cell-free expression system. When this transcription/translation system was exposed to a 72-Hz sinusoidal magnetic field in the range 0.07 to 1.1 mT (rms) for periods of 5 min to 1 h, expression was enhanced. Weaker fields must be applied longer to produce an effect. For 10 min of field exposure, the threshold for an effect is 0.1 mT. These experiments demonstrate that an intact membrane is not an absolute requirement for transducing magnetic bio-effects.

The IDDMK(1,2)22 retrovirus is not detectable in either mRNA or genomic DNA from patients with type 1 diabetes.

A new viral sequence (IDDMK(1,2)22) similar to the human endogenous retrovirus (HERV)-K10/K18 subfamilies has recently been isolated from the culture supernatants of leukocyte-infiltrated islets from two patients who died at the onset of type 1 diabetes. It was claimed that this endogenous retrovirus is expressed in patients with type 1 diabetes but not in healthy control subjects, suggesting an important role of the retrovirus in beta-cell-specific autoimmunity that results in type 1 diabetes. However, despite exhaustive attempts involving identical and expanded methods of detection, we did not observe the IDDMK(1,2)22 viral sequence in genomic DNA, lymphocyte, or plasma RNA in any subject. Therefore, we believe that the viral sequence is not derived from an endogenous retrovirus and that a role for the retrovirus in the pathogenesis of type 1 diabetes must be reconsidered.

Autoreactive diabetogenic T-cells in NOD mice can efficiently expand from a greatly reduced precursor pool.

A broad repertoire of pancreatic beta-cell autoreactive T-cells normally contributes to the development of type 1 diabetes in NOD mice. However, it has been unknown if a large reduction in the precursor pool from which autoreactive T-cells are drawn would inhibit the development of type 1 diabetes. To address this issue, we reduced the precursor frequency of autoreactive T-cells in NOD mice through allelic exclusion induced by transgenic expression of an H2-Db class I-restricted T-cell receptor (TCR) specific for a pathologically irrelevant lymphocytic choriomeningitis virus (LCMV) peptide. TCR allelic exclusion greatly reduced the pool of T-cells from which diabetogenic effectors could be derived in these NODxLCMV TCR Tg mice. Surprisingly, this did not impair their type 1 diabetes susceptibility. Furthermore, a diabetogenic CD8 T-cell population that is prevalent in standard NOD mice was present at essentially equivalent levels in pancreatic islets of NODxLCMV TCR Tg mice. Other data indicated that the antigenic specificity of these CD8 T-cells is primarily the function of a shared TCR-alpha chain. Although the percentage of TCR transgenic T-cells decreased in NOD versus B6,D2 control mice, much higher total numbers of both the TCR transgenic and the nontransgenic T-cells accumulated in the NOD strain. This transgenic T-cell accumulation in the absence of the cognate peptide indicated that the NOD genetic background preferentially promotes a highly efficient antigen-independent T-cell expansion. This might allow diabetogenic T-cells in NOD mice to undergo an efficient expansion before encountering antigen, which would represent an important and previously unconsidered aspect of pathogenesis.

Genetic and physical mapping of a type 1 diabetes susceptibility gene (IDDM12) to a 100-kb phagemid artificial chromosome clone containing D2S72-CTLA4-D2S105 on chromosome 2q33.

Polymorphic markers within the CTLA4 gene on chromosome 2q33 have been shown to be associated with type 1 diabetes. Therefore, a gene responsible for the disease (IDDM12) most likely lies within a region of <1-2 cM of CTLA4. To define more precisely the IDDM12 interval, we genotyped a multiethnic (U.S. Caucasian, Mexican-American, French, Spanish, Korean, and Chinese) collection of 178 simplex and 350 multiplex families for 10 polymorphic markers within a genomic interval of approximately 300 kb, which contains the candidate genes CTLA4 and CD28. The order of these markers (D2S346, CD28, GGAA19E07, D2S307, D2S72, CTLA4, D2S105, and GATA52A04) was determined by sequence tagged site content mapping of bacterial artificial chromosome (BAC) and yeast artificial chromosome (YAC) clones. The transmission disequilibrium test (TDT) analyses of our data revealed significant association/linkage with three markers within CTLA4 and two immediate flanking markers (D2S72 and D2S105) on each side of CTLA4 but not with more distant markers including the candidate gene CD28. Tsp analyses revealed significant association only with the three polymorphic markers within the CTLA4 gene. The markers linked and associated with type 1 diabetes are contained within a phagemid artificial chromosome clone of 100 kb, suggesting that the IDDM12 locus is either CTLA4 or an unknown gene in very close proximity.

Effects of electromagnetic fields on molecules and cells.

Evidence suggests that cell processes can be influenced by weak electromagnetic fields (EMFs). EMFs appear to represent a global interference or stress to which a cell can adapt without catastrophic consequences. There may be exceptions to this observation, however, such as the putative role of EMFs as promoters in the presence of a primary tumor initiator. The nature of the response suggests that the cell is viewing EMFs as it would another subtle environmental change. The age and state of the cell can profoundly affect the EMF bioresponse. There is no evidence that direct posttranscription effects occur as a result of EMF exposure. Although transcription alterations occur, no apparent disruption in routine physiological processes such as growth and division is immediately evident. What is usually observed is a transient perturbation followed by an adjustment by the normal homeostatic machinery of the cells. DNA does not appear to be significantly altered by EMF. If EMF exposure is associated with an increased risk of cancer, the paucity of genotoxic effects would support the suggestion that the fields act in tumor promotion rather than initiation. The site(s) and mechanisms of interaction remain to be elaborated. Although there are numerous studies and hypotheses that suggest the membrane represents the primary site of interaction, there are also several different studies showing that in vitro systems, including cell-free systems, are responsive to EMFs. The debate about potential hazards or therapeutic value of weak electromagnetic fields will continue until the mechanism of interaction has been clarified.

Magnetic field effects on mitotic cycle length in Physarum.

Large plasmodia of Physarum polycephalum were formed from mixtures of micro-plasmodia grown in shaker cultures exposed to 2.0 G (rms), 75 Hz magnetic fields and non-exposed, control cultures. The exposed cultures had been grown continuously in the field and displayed a longer mitotic cycle than the controls. Mixed cultures display synchronous mitosis and a cycle length intermediate to the cycle lengths of exposed and control cultures. The cycle length of mixed cultures varied with the proportions of the mixture in a non-linear manner. The results are discussed in terms of several models.

Low frequency electric and magnetic fields have different effects on the cell surface.

There is a considerable controversy over the nature of weak electromagnetic-field effects in living organisms. Part of the controversy can be traced to a lack of understanding of whether electric or magnetic fields are involved in producing bioeffects. We find that both 60 Hz electric and magnetic fields alter the cell surface of Physarum polycephalum. Exposure to electric fields increases the negative charge on the cell surface while magnetic-field exposure decreases the hydrophobic character of the surface. These effects appear to be additive and independent of the waveform of the applied fields.

Pulsed magnetic fields alter the cell surface.

Pulsed magnetic fields (PMFS) are routinely used in the medical community to facilitate bone repair in clinical cases of non-union or pseudarthoses [(1984) Orth. Clin. No. Am. 15, 61-87]. Although this therapeutic regimen appears to be reasonably effective, the mechanism of action between specific PMFs and the target tissue remains unknown. Adding urgency to the need to understand the mechanism are a wide number of reports that have appeared which demonstrate that PMFs similar to those in clinical use can alter many basic physiological functions. We report that a 24 h exposure to PMFs alters the cell surface of Physarum polycephalum amoebae. Further, using the technique of aqueous two-phase partitioning, we present evidence for individual magnetic and electric field, cell surface effects.

Hearing and neurodevelopmental outcome in survivors of persistent pulmonary hypertension of the newborn.

Thirty-four infants who had a diagnosis of severe persistent pulmonary hypertension of the newborn at birth (alveolar-arterial oxygen difference greater than 600) were treated without paralysis or hyperventilation to induce alkalosis. All survived. Twenty-seven of these 34 eligible infants (79%) underwent neurologic, intelligence, and audiologic testing between 10 months and 6 years of age. Children who were younger than 1 year of age at the initial hearing test were retested after they reached 2 years of age. The average IQ was within the normal range (mean = 96.23). None had sensorineural hearing loss. Severe neurologic abnormalities were seen in 4 children, 3 of whom had been severely asphyxiated at birth (determined by biochemical criteria). Mild neurologic abnormalities were observed in 5 children. Two infants had bronchopulmonary dysplasia because they required supplemental oxygen for 29 and 66 days, respectively, and had abnormal chest roentgenograms; 1 patient takes intermittent doses of albuterol (Ventolin) and neither currently requires supplemental oxygen. This study of 27 infants with severe persistent pulmonary hypertension of the newborn suggests that conservative management without induced alkalosis or respiratory paralysis is accompanied by no sensorineural hearing loss and a good neurologic outcome.

Genetic homogeneity, high-resolution mapping, and mutation analysis of the urofacial (Ochoa) syndrome and exclusion of the glutamate oxaloacetate transaminase gene (GOT1) in the critical region as the disease gene.

The urofacial (Ochoa) syndrome (UFS) is a rare autosomal recessive disorder characterized by abnormal facial expression and urinary abnormalities. Previously, we mapped the gene to a genomic interval of approximately 1 cM on chromosome region 10q23-24, using families from Columbia. Here we demonstrate genetic homogeneity of the syndrome through homozygosity mapping in American patients with Irish heritage. We established a physical map and identified novel polymorphic markers in the UFS critical region. Haplotype analysis using the new markers mapped the UFS gene within one YAC clone of 1,410 kb. We also determined the precise location of the gene encoding for glutamate oxaloacetate transaminase (GOT1) within the new UFS critical region and determined its genomic structure. However, mutation analysis excluded GOT1 as a candidate for the UFS gene.

Chromosomal localization and complete genomic sequence of the murine autoimmune regulator gene (Aire).

We have recently cloned the murine autoimmune regulator (Aire) gene, the homologue of human AIRE responsible for the autoimmune polyglandular syndrome type 1 (APS1) or autoimmune polyendocrinopathy candidiasis ectodermal dystrophy (APECED). Here, we report the genomic sequence (18,413 bp) for the entire Aire gene and its 5' flanking region, which contains putative regulatory sequences. Comparison of the genomic and cDNA sequences indicates that the Aire gene is composed of 14 exons and the coding sequence shares high similarities between mouse and human. The sizes of the homologous introns in the two species are conserved; however, the introns do not share significant sequence homologies except the sequences near the splice donor and acceptor sites. Sequence analyses of the 5' regulatory region and the complete coding region in three mouse strains (B6, NOD and SJL) did not reveal any sequence variation, suggesting sequence conservation between different inbred mouse strains. Using one of the six microsatellite markers identified by genomic sequencing and a B6 x Cast backcross mapping panel, we mapped the mouse Aire gene to chromosome 10, a syntenic region containing the Cdl18 and Pfkl genes on human chromosome 21q22.

Comparison of MB-check and Löwenstein-Jensen media for recovery of mycobacteria.

We have compared the rate of recovery of mycobacteria with the MB-Check culture system (liquid phase) and the Löwenstein-Jensen (LJ) medium in 2,907 clinical specimens obtained from 830 patients submitted for mycobacterial culture during 1-year period. Direct smear examination was carried out by auramine-rhodamine staining. All primary isolates from the culture media were confirmed by Ziehl-Neelsen staining and identified by acridinium-ester-labeled DNA probes specific for Mycobaterium tuberculosis complex. A total of 214 isolates were of the M. tuberculosis complex (88 patients) and 54 of "potentially pathogenic environmental mycobacteria" (45 patients). A total of 117 (54.7%) samples were smear-positive and the remaining 97 (45.3%) were smear-negative. There was a significant difference in the percentage of positive cultures obtained by the MB-Check method (99.1%) as compared with the LJ medium (73.8%) (P < 0.05). This difference, however, occurred almost exclusively at the expense of the 97 smear-negative samples (positive cultures 97.95% by the MB-Check method vs. 42.3% by the LJ culture, P < 0.05). The number of patients diagnosed of tuberculosis by the MB-Check was significantly higher as compared with LJ medium (88 [100%] vs. 77 [87.5%], P < 0.05). In 11 (12.5%) patients, the diagnosis was only established by the MB-Check system. In smear-positive samples, the mean (+/-SD) detection time for M. tuberculosis complex was 14.8 +/- 8 days with MB-Check and 19.9 +/- 7 days with LJ medium. The corresponding figures in smear-negative samples were 22.8 +/- 3 days and 27.8 +/- 6 days, respectively. DNA probes directly applied to MB-Check liquid medium showed a sensitivity of 98.8% and specificity of 100%. These results indicate that the MB-Check system is more efficient for the recovery of mycobacteria than LJ medium.

Long tracheobronchial and esophageal rupture after blunt chest trauma: injury by airway bursting.

Tracheobronchial rupture can be associated with blunt thoracic trauma. An important factor in the physiopathology of these lesions is reflex closure of the glottis, which can be related to closed chest trauma. We report a case of nonpenetrating thoracic trauma that caused a long membranous tracheal rupture from the subcricoid area to the main carina, extending to both main bronchi. In addition, a complex esophageal rupture occurred due to the great energy liberated by the airway rupture acting as a real tracheal burst. Both lesions were diagnosed by flexible bronchoscopy. The postoperative period was without serious complications.

Fimbrial adhesins: similarities and variations in structure and biogenesis.

Fimbriae are wiry (2 to 4 nm diam.) or rod-shaped (6 to 8 nm diam.), fibre-like structures on the surfaces of bacteria which mediate attachment to host cells. Much has been learned in recent years about the biogenesis, structure and regulation of expression of these adhesive organelles in Gram-negative bacteria. Analyses of the genetic determinants encoding the biogenesis of fimbriae has revealed that the adhesive interaction of fimbriae can be mediated by major subunits (CFA/I and CS1 fimbriae) or minor subunits (P, S, and type 1 fimbriae), with the adhesin being located either at the tip of the fimbria or along the length of the fimbrial shaft. Minor subunits can also act as adapters, anchors, initiators or elongators. Post-translational glycosylation of the type 4 pilins of Neisseria gonorrhoeae, Neisseria meningitidis and Pseudomonas aeruginosa has been demonstrated. The structures of the PapD chaperone of Escherichia coli and of N. gonorrhoeae type 4 fimbrin have been resolved at 2.0-2.6 A. Rod-shaped fimbriae should not be thought of as being rigid inflexible structures but rather as dynamic structures which can undergo transition from a helicoidal to a fibrillar conformation to provide a degree of elasticity and plasticity to the fimbriae so that they can resist shear forces, rather like a bungee cord. At least four mechanisms have been identified in the assembly of fimbriae from fimbrin subunits, namely the chaperone-usher pathway (e.g., P-fimbriae of uropathogenic E. coli), the general secretion assembly pathway (e.g., type 4 fimbriae or N-methylphenylalanine fimbriae of P. aeruginosa, the extracellular nucleation-precipitation pathway (e.g., curli of E. coli) and the CFA/I, CS1 and CS2 fimbrial pathway.