Evaluation of the performance of the Helico Blot 2.1 as a tool to investigate the virulence properties of Helicobacter pylori.

Knowledge of the cagA status of Helicobacter pylori infection could be relevant for the treatment and prevention of possible complications of infection. This goal can be achieved using serology since CagA is highly immunogenic. The aim of this study was to correlate the presence of the cagA gene detected by polymerase chain reaction (PCR) with the presence of serum antibodies against the CagA protein using a recently available commercial immunoblotting assay, the Helico Blot 2.1 kit. Considering the results obtained by PCR, Helico Blot 2.1 presented sensitivity, specificity, and positive and negative predictive values for CagA status of 97.4%, 87.7%, 80.9% and 98.5%, respectively.

[How to determine the diagnosis of Helicobacter pylori infection in the elderly?]. / Comment déterminer le diagnostic de l'infection à Helicobacter pylori chez la personne âgée?

PURPOSE: The real prevalence of Helicobacter pylori (H. pylori) infection is difficult to determine in the elderly because of the frequency of drug intake (antibiotics or anti-secretory drugs). The aim of this study was to evaluate the diagnostic performance of five tests in the elderly. METHODS: The study population consisted of consecutive patients undergoing a routine endoscopy between August 1998 and December 1999. We evaluated the diagnostic performance of four tests in all of the included patients: culture and histology of biopsy specimens, serology (ELISA) and urea breath test (13C-UBT). Detection of H. pylori antigens in stool samples (HpSA) was realized in a subgroup. Patients were considered H. pylori + when result for culture was positive or when two tests were positive. RESULTS: One hundred and sixty-seven patients were included in this study (55 men, 112 women; mean age: 85.6 +/- 5.1 years). Only 38 (22.8%) patients were H. pylori+. Test performances showed the following results: serology sensitivity: 90.9% (IC 95%: 75.6-98.1) versus 86.9% (IC 95%: 63.6-96.9) for culture versus 77.8% (IC 95%: 60.8-89.9) for histology and 74.3% (IC 95%: 56.7-87.5) for 13C-UBT. Eighty-nine (53.3%) took antibiotics or anti-secretory drugs, only 13C-UBT performances decreased significantly (sensitivity: 94.4% [72.7-99.8] versus 52.9% [27.8-77]; P < 10(-6)). When gastric or duodenal ulcer were endoscopically diagnosed in older patients, both histology and 13C-UBT could not improve the diagnosis of H. pylori infection. HpSA was realized in 107 patients (sensitivity: 74.1%, specificity: 98.7%). We showed no statistical difference between HpSA performances and drug intake. CONCLUSION: Diagnostic performances decreased in older patients especially because of drug intake.

Magnetic immuno-PCR assay with inhibitor removal for direct detection of Helicobacter pylori in human feces.

A PCR protocol was developed to detect Helicobacter pylori in human stool specimens. This protocol was based on the association of a magnetic immuno-PCR assay with a technique to remove inhibitors (agarose-embedded DNA preparation). Of the 47 H. pylori-positive and 57 H. pylori-negative patients included in this study, 38 were positive and 66 were negative by this new protocol. The sensitivity, specificity, and predictive values for a positive or a negative result were 80.9% (95% confidence interval [CI], 66.3 to 90.4), 100% (95% CI, 92.1 to 100), 100% (95% CI, 88.6 to 100), and 86.4% (95% CI, 75.2 to 93.2), respectively.

Detection of Helicobacter pylori DNA in human feces by PCR: DNA stability and removal of inhibitors.

In this study, the stability of Helicobacter pylori DNA in human feces and the effect of a diet lacking in plant material, the suspected source of PCR inhibitors in human feces, were investigated. In addition, a method to remove these inhibitors was developed. Stools inoculated with H. pylori were used as a model. For this purpose, a H. pylori suspension (10(8) CFU/ml) was used to spike stool samples obtained from four healthy adults known to be H. pylori negative. The evaluation of the stability of H. pylori DNA in feces showed that DNA was degraded after 3 days of contact with fecal material at 37 degrees C. A 2-day diet completely free of plant material was sufficient to eliminate PCR inhibitors from human feces. However, inhibitors were detected 48 h after a normal diet was resumed. A new technique consisting of agarose blocks containing embedded DNA as a template for PCR amplification was used for removal of inhibitors, following DNA extraction by a modified QIAamp tissue method (Qiagen, Hilden, Germany). When this method was applied to inhibiting stool samples known to have an inhibitory effect and spiked with H. pylori (5.10(8) CFU/g), a positive PCR was obtained showing that inhibitors present in the original DNA samples were completely removed. The agarose embedded DNA block method is highly efficient and provides clean, high quality template DNA for PCR purposes avoiding long and fastidious conventional extraction methods. In conclusion, this study confirms that H. pylori DNA degrades with time in stools. A diet free of plant material or a special DNA preparation can be used to remove inhibitors and to allow the detection of H. pylori.

Single and multiple selenium-zinc-iodine deficiencies affect rat thyroid metabolism and ultrastructure.

This study was conducted to evaluate the effects of single and combined deficiencies of Se, Zn and I on thyroid function in rats. Rats were fed amino acid-based diets for 6 wk starting from weaning. The diets contained either low or adequate amounts of these minerals. In addition to the control and control pair-fed groups, seven experimental groups were formed: Se deficient (Se-); I deficient (I-); Zn deficient (Zn-); Se and I deficient (Se-I-); Zn and I deficient (Zn-I-); Se and Zn deficient (Se-Zn); and Se, I and Zn deficient (Se-I-Zn-). Serum triiodothyronine (T3) was significantly lower than in controls in Zn-, Se-Zn- and Se-I- groups. Serum total thyroxine (T4) and free T4 were significantly lower and thyroid-stimulating hormone (TSH) greater in all iodine-deficient groups, regardless of Se or Zn status. Thyroid glutathione peroxidase activity was significantly reduced in Se- and Se-Zn- groups. Nevertheless, in the groups with a concurrent I deficiency, the activity of this enzyme was significantly greater than in controls. Severe alterations of the follicle cellular architecture, including signs compatible with apoptosis, were observed in the Zn- and Se-Zn- groups. These alterations appeared to be less severe when iodine deficiency was simultaneously present. Single and multiple deficiencies of Se, Zn and I have distinct effects on thyroid metabolism and structure.

The effects of breathing pattern training on ventilatory function in patients with COPD.

The purpose of this study was to assess the effects of a particular breathing pattern training (BPT) on forced expiratory volume during the first second (FEV1) and forced vital capacity (FVC) in patients with chronic obstructive pulmonary disease (COPD). The subjects adjusted each breath to a target breath displayed on a video screen, by using visual feedback. This target was chosen in an individual sample recorded at rest. We used a randomized, controlled group design. Twenty patients with stable COPD, FEV1 less than 1.5 liters, undergoing a traditional rehabilitation program were randomly assigned to the BPT group or to the control group. Each BPT subject underwent 30-35 training sessions spread out over four weeks, in addition to the traditional program. FEV1 and FVC were performed before and after this program. ANOVAs showed that FEV1 and FVC significantly improved in BPT subjects, with a mean percent increase of 22% and 19%, respectively. Corresponding changes in controls were not significant. This study showed short-term increases in FEV1 and FVC in COPD patients practicing BPT in addition to respiratory rehabilitation, in comparison with controls. Further studies should incorporate outcome data to clarify the mechanisms and the duration of this effect.

Voluntary control of breathing pattern in asthmatic children.

15 asthmatic children and 15 healthy children were trained to adjust their breathing pattern to a target pattern displayed on a video screen by using visual feedback. The error scores in the two groups were not significantly different. These data did not support the hypothesis that voluntary control of respiratory muscles is impaired in asthmatics.

Variability of cadmium, copper and zinc levels in molluscs and associated sediments from Chile.

The concentrations of cadmium, copper and zinc in mussel and sediment samples collected together from eight different geographical coastal areas of Chile were determined. The mussels studied were 'Chorito Maico', 'Almejas' and 'Navajuelas Chilenas' (Perumytilus purpuratus, Semelle solida and Tagellus dombeii, respectively). Sampling was carried out in July and September 1992 and January and April 1993 (winter, spring, summer and autumn seasons in Chile, respectively). The metal levels in these mussels varied among species; there were several sites where the metal concentrations in molluscs approached or exceeded the criteria levels for Cd, Cu and Zn in shellfish products: 1, 10 and 50 ppm ww respectively, which are regarded as safe levels for human consumption. The results of metal levels in sediments showed two areas clearly polluted with Cu. Strong relationships between Cu concentrations in the three molluscs and sediments were found; weak correlations were observed for Zn in S. solida.

[Mercury in the hair of pregnant and lactating Chilean mothers]. / Mercurio en el cabello de embarazadas y madres lactantes chilenas.

Mercury-containing industrial waste has been released into the coastal waters of the Eighth Region of Chile for around two decades. This study, carried out from 1991 to 1993, sought to measure mercury concentrations in the hair of pregnant and lactating women from villages near the coast and in the interior of the region in order to examine the relationship between the concentration of mercury and seafood consumption. The survey questionnaire used in 1991 to determine seafood consumption did not ask about the frequency of consumption of fish, shellfish, and algae but only whether the women who were pregnant or breast-feeding consumed a minimum of one fish-based meal per week. The questionnaire used in 1992 and 1993 asked about the daily and weekly consumption of seafood in general (fish, shellfish, and algae). Spectrophotometry was used to determine the total mercury concentration in samples of 100 mg of hair from 153 pregnant and lactating women in 11 fishing villages of the Eighth Region where seafood is regularly consumed. None of the women had occupational exposure to mercury. Total mercury concentration was also determined in hair samples from a control group composed of 26 pregnant and lactating women from Pinto and El Carmen, villages in the interior of the same region where seafood was rarely eaten. The arithmetic mean of the total mercury concentration in hair was 1.81 mg/kg of body weight for the study group (standard deviation [SD] 1.52) and 0.42 mg/kg for the control group (SD 0.15)--a statistically significant difference (P < 0.01). Pairwise comparisons also revealed statistically significant differences (P < 0.05) between the mean for the interior group and the means for the women in the nine villages closest to the sources of the pollution, but not between the mean for the interior group and those for women in the two villages at the extreme north and south of the study zone, who lived farthest from the contaminated waters. The total mercury concentration in hair was significantly higher in women who indicated that they ate fish seven or more times per week; in those who said they ate fish, shellfish, or algae five or more times per week; and in those who had lived 20 or more years in their village. No statistically significant differences were found when the results were analyzed by age.

Characterization of the regional distribution of selenium in Chile using selenium in hens' eggs as a monitor.

Regional distribution of selenium (Se) in Chile was investigated by using the concentration of Se in hens' eggs as a monitor. Forty-one locations along the entire length of the country were sampled. Average (+/- SD) egg-white Se content (mg/kg dry) was 0.79 +/- 0.41, range 0.22-2.23. Corresponding yolk Se values were, mean 0.81 +/- 0.43, (mg/kg dry) range 0.26-2.23. Locations grouped in five main areas, according to their geographic-climatic characteristics, showed significant differences regarding both egg-white Se and yolk Se. Analyzed dietary Se intake from two distinct areas reflected the trends observed in the Se content of egg fractions from such regions. These data support the utilization of the concentration of Se in hens' eggs as a useful monitor of dietary selenium consumed by selected populations.

Determination of total mercury in scalp hair of pregnant and nursing women resident in fishing villages in the Eighth Region of Chile.

Total mercury (Hg-T) content in scalp hair of 59 pregnant and nursing women-with normal to high fish and seafood consumption-resident in fishing villages distributed throughout the coastal zone of the Eighth Region of Chile, and of 7 pregnant and nursing women-with negligible or no fish and seafood consumption-resident in a town located inland (Pinto) in the same region, was determined. Hair samples were collected, washed and stored according to IAEA recommendations, and were wet-digested in sealed Pyrex ampoules prior to the determination of Hg-T by gold amalgamation cold-vapour atomic absorption spectrometry. The absolute detection limit was estimated as 0.13 +/- 0.01 ng Hg-T (3 x sigma B1). Accuracy and precision were assessed with the aid of various biological and environmental certified and standard reference materials including human hair, and were satisfactory for ppm and sub-ppm Hg-T. Instrumental neutron activation analysis was used as a reference method for external quality control. The Hg-T content in hair of the study group (2.06 +/- 1.45(8) mg/Kg) was significantly higher than that of the control group (0.43 +/- 0.18(4) mg/kg) (p = 0.0001). These results were characterized according to geographical location, fish and seafood consumption, age and residence period in the fishing village.

Cadmium, lead, copper and mercury levels in fresh and canned bivalve mussels Tagelus dombeii (Navajuela) and Semelle sólida (Almeja) from the Chilean coast.

Samples of bivalve mussels Tagelus dombeii and Semelle sólida (Navajuelas and Almejas chilenas, respectively) caught at different sites along the coast of Chile were analysed for Cu, Cd, Hg and Pb content. The analyses were performed on samples of fresh and canned products, classified according to their length. Each Navajuela sample was divided into two subsamples. One was dissected and the visceral tissue, branchial tissue and gonads were removed, while the other sample consisted of the whole tissues. The heavy metal concentration of fresh mussel samples was compared with the same samples as canned products. The data show that for both species there are no significant differences in the metal concentration with the mollusc size in fresh samples and canned products. It has been shown that there is no contribution to the trace metal content (Cd, Pb and Hg) in canned Navajuelas and Almejas mussels as a consequence of the industrial process. A clear and significant reduction in the Cd and Cu content was observed in Navajuelas without visceral tissue and Almejas without mantle tissue for both the fresh and canned products. This is consistent and is corroborated with the high metal concentrations found in the visceral and mantle tissue.

Performance and learning during voluntary control of breath patterns.

Fourteen subjects learned to adjust their breath pattern to two target breaths displayed on a video screen, by using visual feedback, during two sessions 24 h apart. These two targets were respectively the smallest and the largest breaths of a ten-breath sample previously recorded from each subject's resting spontaneous breathing. Performances were significantly better for the large than for the small target breath. This cannot be directly inferred from current knowledge related to the control of movement time and amplitude, but rather it may be inferred from the periodic character of breathing, to the higher mental load during the small breath task, or to the presumably different frequencies of target breaths in the whole span of spontaneous breathing. In the second session, performance on the two targets levelled out as a result of learning.

[Concentration of trace elements in children from northern Chile]. / Concentración de elementos traza en niños del norte de Chile.

Industrial processes may lead to severe alterations of environment affecting the health and quality of life of neighbouring communities. The present study reports the concentrations of Cu, As, Br, Na, K, Se, Hg, Cr, Co, Zn, Fe and Sb in samples of hair from children 6 years of age living in Antofagasta, Chuquicamata, María Elena, Calama and San Pedro de Atacama. Results were compared to those obtained in children of the same age living in Santiago. Samples were obtained according to recommendations from the International Nuclear Energy Organization and were analyzed by the neutron activation method at facilities of the Chilean Nuclear Energy Commission. Values for Cu (38.8 ppm), Cr (4.29 ppm) and As (19.2 ppm) in children from Chuquicamata are considered dangerous according to international standards.

Toxic trace elements in Chilean seafoods: development of analytical quality control procedures.

Chile is a well known producer and exporter of shell fish. These seafoods, like other specimens of marine origin, are susceptible to environmental and other contamination by trace elements, including toxicants. Therefore, adequate analytical quality assurance is mandatory before accepting analytical results. In this context, the use of at least two independent methods of determination and validation with certified reference materials (CRM) provides acceptable criteria for judging the reliability of the data. This paper describes sample treatments and analytical procedures for Cd, Cu and Hg determinations in mollusc samples. Three independent analytical techniques, namely differential pulse anodic stripping voltammetry, neutron activation analysis and atomic absorption spectrometry, were used. CRM standards of the IAEA, NIST and BCR were analyzed to evaluate quality assurance. Following the quality control phase, the concentrations of cadmium, copper and mercury in fresh and canned mollusc samples Tagelus dombeii and Semelle solida (Navajuelas and Almejas chilenas respectively) from different locations were determined.

[Analysis of some iatrogenic injuries during manual endodontic treatment]. / Analyse de certains dégats iatrogéniques au cours des traitements endodontiques manuels.

Some of various iatrogenic diseases, during manual endodontic treatments, are analysed, from detailed clinical instances. Discussion leads to insist upon some classical and original ways of prevention. From general point of view, it is unavoidable to realize with gentleness, never forcing, endodontic proceedings. Deep perception of internal dental morphology is quite necessary and must be respected during treatment. Besides, histo-pathological modifications of parietal dentine have to be known, not only from intellectual but also from tactile point of view. Each practitioner will progressively improve.