Bio monitoring with stable isotopes

The hygiene hypothesis postulates that the increase in atopic diseases may in part be due to diminished exposure to microorganisms. But it is unknown which type of infection does render protection. An epidemiological study was conducted in Leipzig, Germany, and its rural county, involving 3347 school starters. Two types of infection were considered: (1) gastrointestinal colonisation (Helicobacter pylori detection using in vivo [(13)C] urea breath test) and (2) respiratory infections (physician-diagnosed lower (bronchitis) and upper (common cold) respiratory infections). H pylori colonisation was selected because it is very common and plays an important role in gastrointestinal disorders. Atopic eczema was selected as the (allergic) target variable because of its high frequency in the age of the study participants. The results, adjusted for relevant confounders, showed a significant inverse association between H pylori infection and eczema (adjusted odds ratio (aOR) = 0.31, p = 0.006) in children not predisposed to atopy. In contrast, bronchitis increased the risk of eczema (aOR = 1.98, p<0.001). Bacterial digestive tract colonisation (infection) seems to protect against eczema in comparison with the effect of respiratory tract infections. The hygiene hypothesis may be better explained when gastrointestinal and respiratory infections are subtly differentiated.

Conventional diagnosis of the pulmonary tract uses physical methods such as spirometry and oscillometry. However, the inhalation of a chemical diagnostic agent ought to provide novel ways of more specific diagnosis, for instance of inflammatory states of the bronchial and lung mucosa. The stable isotope technique using a (15)N-labeled substrate appears to be a suitable tool for this application. In a pilot study, defined amounts of the amino acid L-[guanidino-(15)N(2)]arginine monohydrochloride (aqueous solution, 20 atom % (15)N) were inhaled as a diagnostic agent by healthy volunteers and pulmonary patients suffering from asthma bronchiale. The amino acid is resorbed and partly metabolized to (15)NO. The exhaled air was collected under defined conditions in 10-L breath bags and analyzed for NO using chemiluminescence. Under standardized test conditions, healthy persons (n = 6) exhaled 0.97 +/- 0.08 micromol NO/m(3) and asthmatic patients (n = 7) 1.17 +/- 0.14 micromol NO/m(3). A better distinction was expected comparing the (15)NO exhalation. The (15)N abundance of NO was determined using a Cryotrap gas chromatography - mass spectrometry set-up. Between 30 and 80 minutes after inhaling 700 mg [(15)N]arginine, a maximum with a plateau of the (15)NO abundance was found in the exhaled air. At this time, healthy and asthmatic subjects exhibited clear differences in their exhaled (15)NO amounts. Under standardized test conditions, the healthy persons (n = 6) exhaled 102.3 +/- 6.7 nmol (15)NO/m(3), whereas asthmatic patients (n = 7) exhaled only 76.1 +/- 10.9 nmol (15)NO/m(3). It is concluded that (15)NO yielded after the inhalation of (15)N-labeled arginine could be a potential marker for demonstrating pathophysiological changes in the lung epithelium. This method could pave a new diagnostic principle of "inhalative breath test."

The [15N]methacetin urine test for assessing disturbances of the cytochrome P450-aided oxygenation of xenobiotics in the human liver has been approved in several environmental health studies. A recent longitudinal study of kindergarten children near chemical and mining companies undergoing fundamental restructuring showed high, seasonally fluctuating SO2 burden which was well correlated with alterations in the mean liver states of the children. At one point the correlation disappeared, together with indications of large amounts of chlorinated compounds overflowing locally at nighttime. This was finally proven by analyzing urine samples from the children for trichloroacetic acid (TCA). Chlorinated ethanes and ethenes-the precursors of TCA-seemed to dominate the air pollution and to affect the hepatic detoxification capacity. We concluded that the methacetin liver function test reflects multicomponent exposure, including acute monopolization by a dominant pollutant for a time.

Helicobacter pylori (H. pylori) is one of the world's most widespread microorganisms. Its acquisition in humans remains poorly understood, however, epidemiological studies have identified drinking water as reservoir for the bacterium. The aim of this study was to investigate the prevalence of H. pylori infection among individuals using or drinking previously H. pylori tested well water. Applying household cluster sampling, a total of 91 subjects, all using or drinking well water (13 of either H. pylori positive or negative wells), were screened for their H. pylori status. The group was comprised of 73 adults and 19 children under the age of 18. H. pylori infection was determined using the [13C]urea breath test. A self-administered or parent-completed questionnaire provided information on living conditions and lifestyle habits including the use or drinking of well water. Logistic regression analyses associated the drinking of H. pylori positive well water with a positive colonization status [Odds Ratio (OR) 8.3; 95% confidence interval (95% CI) 2.4-29]. In summary, the use or drinking of H. pylori contaminated well water appears associated with the acquisition of a H. pylori infection. This study is based on a relatively small and inhomogeneous population sample and should be repeated to confirm the results.

The possible transmission of Helicobacter pylori (H. pylori) among asymptomatic individuals using the contaminated well waters was investigated. It was found that drinking or using H. pylori contaminated well water is a risk in the acquisition of H. pylori. Age was also found to be a very important risk factor for a positive colonization status. The importance of making the environment safer for children by preventing childhood-acquired illnesses with a high cost-benefit effects was also suggested.

The classical way to demonstrate the efficiency of remediation is measuring the reduction of toxic compounds in the environment. Nevertheless, more important is the risk reduction in human health. To determine changing health effects, exposure and bio-effects have to be monitored at time of and during remediation. Kindergarten children from a heavily polluted industrial (n=23) and a control area (n=12) were investigated. The region-specific outdoor and indoor exposure [27 volatile organic compounds (VOC), emphasis on tri- and tetrachloroethylene (TRI, TETRA)], the internal load [(trichloroacetic acid-TCA-as urine metabolites of TRI and TETRA and S-phenyl- and S-benzylmercapturic acid (SPMA and SBMA) as metabolites of benzene and toluene], and biological effect assessment ([(15)N]methacetin test-a non-invasive stable isotope test to determine the unspecific liver detoxification capacity of an individual) were measured twice a year during 2 years of remediation (1997/1998). It could be shown that in- and outdoor levels of TRI and TETRA decreased by 47% in the heavily polluted village, Greppin, while the levels remained much the same in the control village, Roitzsch. This trend was reflected in the decreasing elimination of TCA in the urine (41%) by the Greppin children, with no differences in the TCA elimination in Roitzsch probands. As the remediation efforts decreased the burden of exposure, the children's liver detoxification capacity improved as well. Combining different methods, such as exposure-effect (external and internal loads) and bio-effect monitoring, proved to be useful to assess remediation successes including the improvement in human health.