[Maternal exposure to dust particles during pregnancy and fetal growth in 2 regions with different levels of air pollution]. / Expozice matky prachovým cásticím tehotenství a rust plodu ve dvou regionech s rozdílnou úrovní znecisteného ovzdusí.

BACKGROUND: Recently, an association between adverse pregnancy outcome and air pollution was hypothesized in several papers. The aim of the present study is to evaluate the impact of particles with aerodynamic diameter < or = 10 microns (PM10) on the intrauterine growth retardation (IUGR) in two regions with the different levels of air pollution. METHODS AND RESULTS: The study group includes all singleton full term births of European origin over a two-year period in the district of Teplice (n = 1,943) and Prachatice (n = 883). Information on reproductive and health history and lifestyle was obtained from maternal questionnaires and medical records. IUGR was chosen as the indicator of reproductive effects. The mean concentration of PM10 for each women in particular gestational month was calculated. Three concentration intervals were constructed (low < 40 micrograms/m3; medium 40-50 micrograms/m3; high > or = 50 micrograms/m3). Relative risk of IUGR was estimated for particular month and pollutant level. Influence of the potential confounders (maternal age, height and pre-pregnancy weight, smoking, season and the year of study) was controlled using logistic regression models; adjusted odd's ratio (AOR) was estimated. Relative risk of IUGR in Teplice District was increased for medium PM10 levels (AOR = 1.62 CI: 1.07-2.46) (P < 0.02) as well as for high levels in the first gestational month (2.64 CI: 1.48-4.71) (P < 0.001). The same association was observed also in Prachatice District, though the PM10 did not achieve the "high" levels (> or = 50 micrograms/m3). AOR for IUGR for mothers exposed in the 1st gestational month to medium PM10 levels was three times the risk of those exposed to low levels (3.50 CI: 1.82-6.81) (P < 0.0003). No similar relationship was observed in any other gestational stage. CONCLUSIONS: Influence of particles or other associated air pollutants on fetal growth in early gestation is one of several possible explanations of these results. Timing of this effect in both different regions is compatible with a current hypothesis of IUGR etio-pathogenesis. Seasonal factors, one of the other possible explanations, is less probable. More investigation is required to evaluate a possible causal nature of the relationship observed.

Genotoxicity of urban air pollutants in the Czech Republic. Part I. Bacterial mutagenic potencies of organic compounds adsorbed on PM10 particulates.

As part of a long-term program to investigate the impact of air pollution on the health of a population in a polluted region in Northern Bohemia, mutagenicity of extractable organic matter (EOM) from air particles PM10 was investigated by the means of Salmonella typhimurium indicator strains TA98 and YG1041 using the Ames plate incorporation assay. The air samples were collected in both the polluted and the control districts during the summers and winters of 1993-1994. In the polluted district, the collection was repeated during the winter of 1996-1997. The crude extracts from filters pooled according to the locality and the season were fractionated by acid-base partitioning into acid, base, and neutral fractions. The neutral fractions were further fractionated by silica gel column chromatography into five subfractions. The induction of revertants with the crude extracts was higher in winter samples than in summer samples. Both indirect-acting and direct-acting mutagenicity were observed. The indirect mutagenic potency of aromatic subfractions containing polycyclic aromatic hydrocarbons (PAHs) was generally low. The mutagenic potency detected with TA98 was more distinct only in the winter sample 1993-1994 from the polluted area, where the aromatic subfraction accounted for 23% of total mutagenicity. In both strains, the highest direct-acting mutagenicity was found in slightly polar fractions containing nitro-PAHs. The mutagenic potency detected with YG1041 was about two orders of magnitude higher than that detected with TA98. No substantial locational- or time-related variances in the mutagenic potencies of EOM, or in the spectrum of chemical components identified in individual fractions were found. The polluted district, in comparison to the control district, was found to have higher amounts of EOM, carcinogenic PAHs and mutagenicity of air particles (rev/m(3)). The fractionating process, combined with the bacterial mutagenicity test, confirmed that nitro-derivatives are the most important contributors to the bacterial mutagenicity of air particles. However, this study did not fulfill the expectancy to bring substantially new, clear-cut information on the composition and the biological activity of air pollution in both districts.

Fecundability and parental exposure to ambient sulfur dioxide.

Recently it has been observed that birth rates in Teplice, a highly polluted district in Northern Bohemia, have been reduced during periods when sulfur dioxide levels were high. This study, which is based on data from 2,585 parental pairs in the same region, describes an analysis of the impact of SO(2) on fecundability in the first unprotected menstrual cycle (FUMC). We obtained detailed personal data, including time-to-pregnancy information, via maternal questionnaires at delivery. We estimated individual exposures to SO(2) in each of the 4 months before conception on the basis of continual central monitoring. Three concentration intervals were introduced: < 40 microg/m(3 )(reference level); 40-80 microg/m(3); and [greater than or equal to] 80 microg/m(3). We estimated adjusted odds ratios (AORs) of conception in the FUMC using logistic regression models. Many variables were screened for confounding. AORs for conception in the FUMC were consistently reduced only for couples exposed in the second month before conception to SO(2) levels as follows: 40-80 microg/m(3), AOR 0.57 [95% confidence interval (CI), 0.37-0.88; p < 0.011]; [greater than or equal to] 80 microg/m(3), AOR 0.49 (CI, 0.29-0.81; p < 0.006). The association was weaker in the second 2 years of the study, probably due to the gradual decrease of SO(2) levels in the region. The relationship between SO(2) and fecundability was greater in couples living close to the central monitoring station (within 3.5 km). The timing of these effects is consistent with the period of sperm maturation. This is in agreement with recent findings; sperm abnormalities originating during spermatid maturation were found in young men from Teplice region who were exposed to the increased levels of ambient SO(2). Alternative explanations of our results are also possible.

The impact of polycyclic aromatic hydrocarbons and fine particles on pregnancy outcome.

The relationship between intrauterine growth retardation (IUGR) and exposure to particulate matter [less than/equal to] 10 microm (PM(10)) and particulate matter [less than and equal to] 2.5 microm (PM(2.5))( )in early pregnancy was recently studied in the highly polluted district of Teplice (Northern Bohemia). From this observation rose the question about the possible role of the carcinogenic fraction of polycyclic aromatic hydrocarbons (c-PAHs), which are usually bound to fine particles. The impact of c-PAHs and fine particles on IUGR was analyzed in Teplice and in Prachatice, a region with similarly high c-PAH but low particle levels. All European, single live births occurring in a 4-year period in Teplice (n = 3,378) and Prachatice (n = 1,505) were included. Detailed personal data were obtained via questionnaires and medical records. Mean PM(10), PM(2.5,) and c-PAHs levels during the 9 gestational months (GM) were estimated for each mother. Adjusted odds ratios (AORs) of IUGR for three levels of c-PAHs (low, medium, and high) and for continuous data were estimated after adjustment for a range of covariates using logistic regression models. In the present 4-year sample from Teplice, previously published results about increasing IUGR risk after exposure to particles in the first GM were fully confirmed, but no such effects were found in Prachatice. The AOR of IUGR for fetuses from Teplice exposed to medium levels of c-PAHs in the first GM was 1.60 [confidence interval (CI), 1.06-2. 15], and to high levels 2.15 (CI, 27-3.63). An exposure-response relationship was established by analyzing the continuous data. For each 10 ng increase of c-PAHs in the first GM, the AOR was 1.22 (CI, 1.07-1.39). About the same relationship was observed in Prachatice in spite of the low particle levels. The results prove that exposure to c-PAHs in early gestation may influence fetal growth. The particulate matter-IUGR association observed earlier may be at least partly explained by the presence of c-PAHs on particle surfaces.

Fetal growth and maternal exposure to particulate matter during pregnancy.

Prior studies reported an association between ambient air concentrations of total suspended particles and SO2 during pregnancy and adverse pregnancy outcomes. We examined the possible impact of particulate matter up to 10 microm (PM10) and up to 2.5 microm (PM2. 5) in size on intrauterine growth retardation (IUGR) risk in a highly polluted area of Northern Bohemia (Teplice District). The study group includes all singleton full-term births of European origin over a 2-year period in the Teplice District. Information on reproductive history, health, and lifestyle was obtained from maternal questionnaires. The mean concentrations of pollutants for each month of gestation were calculated using continuous monitoring data. Three intervals (low, medium, and high) were constructed for each pollutant (tertiles). Odds ratios (ORs) for IUGR for PM10 and PM2.5 levels were generated using logistic regression for each month of gestation after adjustment for potential confounding factors. Adjusted ORs for IUGR related to ambient PM10 levels in the first gestational month increased along the concentration intervals: medium 1.62 [95% confidence interval (CI), 1.07-2.46], high 2.64 (CI, 1.48-4.71). ORs for PM2.5 were 1.26 (CI, 0.81-1.95) and 2.11 (CI, 1. 20-3.70), respectively. No other associations of IUGR risk with particulate matter were found. Influence of particles or other associated air pollutants on fetal growth in early gestation is one of several possible explanations of these results. Timing of this effect is compatible with a current hypothesis of IUGR pathogenesis. Seasonal factors, one of the other possible explanations, is less probable. More investigation is required to examine these findings and alternative explanations.

Risk assessment of the common air pollutants in Teplice, Czech Republic.

In the framework of the systematic investigation of the environment of the district of Teplice (Northern Bohemia), one of the most polluted regions in Europe, an attempt was made to estimate health risks to the inhabitants posed by the most common air contaminants (SO2, NOx, particulate matter). A meta-analysis of data published in recent papers dealing with health effects was performed. At first we weighed the number of positive and negative findings focusing on the following health indicators: prevalence of symptoms (coughing, wheezing), decreased respiratory function, prevalence of respiratory illness, and acute mortality. Only those categories in which the positive findings prevailed were taken into consideration and median values for LOAELs were calculated from the data referring to positive dose-response relationships. The exposure assessment was based upon a series of data on daily concentrations of the air contaminants in Teplice since 1975. Due to the somatic and respiration parameters, as well as to their habits, children between the ages of 8 and 10 appeared the most heavily exposed of all age groups. It was concluded that in real concentrations the risk is posed mainly from sulphur dioxide and, above all, from particulate matter.

Genotoxicity of coke-oven and urban air particulate matter in in vitro acellular assays coupled with 32P-postlabeling and HPLC analysis of DNA adducts.

This study is an in vitro part of the ongoing biomarker studies with population from a polluted region of Northern Bohemia and coke-oven workers from Czech and Slovak Republics. The aim of this study is to compare DNA adduct forming ability of chemical compound classes from both the urban and coke-oven extractable organic mass (EOM) of airborne particles. The crude extracts were fractionated into seven fractions by acid-base partitioning and silica gel column chromatography. In in vitro acellular assays we used calf thymus DNA (CT DNA) with oxidative (+S9) and reductive activation mediated by xanthine oxidase (+XO) under anaerobic conditions. Both the butanol and nuclease P1 versions of 32P-postlabeling for detection of bulky aromatic and/or hydrophobic adducts were used. The results showed that the spectra of major DNA adducts resulting from both the in vitro assays are within the fractions similar for both the urban and coke-oven samples. The highest DNA adduct levels with S9-activation were detected for the neutral aromatic fraction, followed by slightly polar and acidic fractions for both samples. With XO-mediated metabolism, the highest DNA adduct levels were detected for both the acidic fractions. Assuming additivity of compound activities, then the acidic fraction, which in the urban sample comprises a major portion of EOM mass (28%), may contain the greatest activity in both in vitro assays (39 and 69%, +S9 and +XO, respectively). In contrast, the aromatic fraction constituting only 8% of total urban EOM mass may account for comparable activity (34%) with organic acids. The highest DNA adduct forming activity of the coke-oven sample accounts for the aromatic fraction (82 and 63%, +S9 and +XO, respectively) that also contains the greatest portion of the total EOM (48%). To characterize some of the specific DNA adducts formed, we coupled TLC on 20x20 cm plates with HPLC analysis of 32P-postlabeled adducts. In both S9-treated samples of the aromatic fraction, we tentatively identified DNA adducts presumably diolepoxide-derived from: 9-hydroxy-benzo[a]pyrene (9-OH-B[a]P), benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide[+/-] (anti-BPDE), benzo[b,j,k]fluoranthenes (B[b]F, B[j]F, B[k]F), chrysene (CHRY), benz[a]-anthracene (B[a]A) and indeno[cd]pyrene (I[cd]P). These DNA adducts accounted for about 57% of total DNA adducts detected in both S9-treated samples of the aromatic fraction. DNA adducts of XO-treated samples were sensitive to nuclease P1 and HPLC profiles of the major adducts were markedly different from the major adducts of S9-treated samples. However, the combination of TLC and HPLC did not confirm the presence of DNA adducts derived from 1-nitropyrene (1 NP), 9-nitroanthracene (9 NA) and 3-nitrofluoranthene (3 NF) that were detected by GC-MS in the slightly polar fraction. We concluded that the chemical fractionation procedure facilitates the assessing of DNA adduct forming ability of different chemical compound classes. However, based on the results obtained with the whole extracts, it does not fulfil a task of the actual contribution of individual fractions within the activity of the whole extracts. Our results are the first in detecting of DNA adducts derived from urban air and coke-oven particulate matter.

DNA adducts in human placenta as related to air pollution and to GSTM1 genotype.

DNA adducts in human placenta have been studied in relation to metabolic genotype for glutathione S-transferase M1 (GSTM1) in 98 mothers living in two regions with a different annual average air pollution levels: Northern Bohemia-the district of Teplice as polluted industrial area (mines, brown coal power plants) and Southern Bohemia-the district of Prachatice as agricultural area without heavy industry. Forty-nine placenta samples (25 from the Teplice district and 24 from the Prachatice district) from non-smoking mothers with the date of delivery in the summer period and 49 placenta samples (25 from the Teplice district and 24 from Prachatice district) from mothers with the date of delivery in the winter period were analysed. The total DNA adduct levels were calculated as the sum of adducts in the diagnoal radioactive zone (DRZ) and one distinct spot outside of the DRZ (termed X), which was detected in almost all placenta samples. We found total DNA adduct levels of 1.40 +/- 0.87 (0.04-3.65) and 1.04 +/- 0.63 (0.11-3.08) adducts per 10(8) nucleotides for the Teplice and Prachatice districts, respectively. The significant difference between both districts in placental DNA adduct levels was found for the winter sampling period only (1.49 vs. 0.96 adducts per 10(8) nucleotides; p = 0.023). No seasonal variation was observed for DNA adduct levels in the overall population studied. A positive GSTM1 genotype was detected in 51 subjects, while GSTM1-null genotype was found in 47 subjects. Higher DNA adduct levels were detected in a group with GSTM1-null genotype (p = 0.009). This finding seems more significant for subjects in the Teplice district (p = 0.047) than for those in the Prachatice district (p = 0.092). Significant district and seasonal differences were found in subgroups carrying the GSTM1-null genotype. DNA adduct levels in placentas of mothers with GSTM1-null genotype living in the polluted district of Teplice were higher than those in Prachatice (p = 0.050); also the adduct levels in placentas sampled in the summer period were higher than those sampled in the winter period (p = 0.011). Our results indicate that simultaneous analysis of DNA adducts and metabolic genotypes could emphasize the use of DNA adduct measurements, particularly in the case of the environmental exposure when the total doses of genotoxic pollutants are very low.

Characterization of phosphate transport in Streptomyces granaticolor.

Transport of inorganic phosphate in Streptomyces granaticolor was characterized in two growth stages; kinetic parameters were determined and two transport systems were found in both stages, with the following values: KT1 = 0.06 mM, Jlim1 = 0.95 nmol min-1 (mg DS)-1, and KT2 = 1.80 mM, Jlim2 = 25 nmol min-1 (mg DS)-1. Both systems require metabolic energy and substrates, such as sugars or polyols; when alanine was used or the energy source was omitted, the kinetic parameters changed in both systems. Both systems were inhibited by the ionophore cccp with identical k(i). KCN, an inhibitor or terminal cytochrome oxidase, inhibited the uptake of phosphate only partially, the uptake was inhibited completely when also the inhibitor of the alternative oxidative pathway (salicylhydroxamic acid) was added. Antimycin A inhibited the uptake completely. Arsenate inhibited competitively.

Cell volume regulation in Claviceps fusiformis. An animal-type Na,K-ATPase operating in a fungus?

Claviceps fusiformis, an ergot pathogen of pearl millet, was equilibrated in media without and with 0.1 mM ouabain (g-strophantin). Highly significant effects of ouabain on the contents of sodium ions and water were found, suggesting that an active transport of sodium ions, similar to that in animal cells, regulates the cell volume of the fungus. The basic role of the sodium pump appears to be to decrease larger cell volumes and to increase the small ones; for this purpose it can even reverse its direction.

Influence of GSTM1 and NAT2 genotypes on placental DNA adducts in an environmentally exposed population.

The placenta bulky DNA adducts have been studied in relation to metabolic genotypes for glutathione S-transferase M1 (GSTM1) and N-acetyl transferase 2 (NAT2) in 158 mothers (113 nonsmokers and 45 smokers) living in two regions with different annual average air pollution levels of sulphur dioxide, nitrogen oxides, particulate matter < 10 microns, and polycyclic aromatic hydrocarbons. One region was the district of Teplice as the polluted industrial region with mines and brown coal power plants, and the other was the district of Prachatice, an agricultural region without heavy industry. DNA adduct levels were determined by using a butanol extraction enrichment procedure of 32P-postlabeling. GSTM1 and NAT2 genotypes were studied by using polymerase chain reaction. The total DNA adduct levels included a diagonal radioactive zone (DRZ) and one distinct spot outside DRZ (termed X), which was detected in almost all placenta samples and correlated with DRZ (r = .682; P < .001). We found the total DNA adduct levels 2.12 +/- 1.46 (0.04-7.70) and 1.48 +/- 1.09 (0.11-4.98) adducts per 10(8) nucleotides for Teplice and Prachatice districts, respectively, indicating significant differences between both regions studied (P = .004). Elevated DNA adduct levels were found in smoking mothers (10 or more cigarettes per day) by comparison with nonsmoking mothers (3.21 +/- 1.39 versus 1.32 +/- 0.88 adducts per 10(8) nucleotides; P < .001). Placental DNA adduct levels in smokers correlated with cotinine measured in plasma (r = .432; P = .003). This relation indicates that cigarette smoking could be predominantly responsible for DNA adduct formation in placentas of smoking mothers. DNA adduct levels were evaluated separately for non-smokers (1.50 +/- 1.00 vs. 1.09 +/- 0.66 adducts/10(8) nucleotides for the Teplice and Prachatice districts, respectively; P = .046) and smokers (3.35 +/- 1.47 vs. 2.91 +/- 1.20 adducts/10(8) nucleotides for Teplice and Prachatice districts, respectively; P = .384) to exclude the effect of active cigarette smoking on the district variation. These findings indicate that the effect of the environmental pollution in cigarette smokers is practically overlapped by tobacco exposure. No seasonal variation was observed for DNA adduct levels in the overall population studied and no relation between total DNA adduct levels in placenta and levels of vitamins A, C, and E in venous and cord blood was found. A positive GSTM1 genotype was detected in 78 subjects, while negative GSTM1 genotype was found in 80 subjects. Higher DNA adduct levels were detected in the group with GSTM1-negative genotype by comparison with GSTM1-positive genotype (2.05 +/- 1.30 vs. 1.66 +/- 1.39 adducts/10(8) nucleotides; P = .018). This finding is more pronounced in the Teplice district (2.33 +/- 1.36 vs. 1.88 +/- 1.56 adducts/10(8) nucleotides; P = .053) than for the Prachatice district (1.61 +/- 1.09 vs. 1.36 +/- 1.10 adducts/10(8) nucleotides; P = .248) and for nonsmokers (1.45 +/- 0.82 vs. 1.18 +/- 0.93 adducts/10(8) nucleotides; P = .029) more than for smokers (3.45 +/- 1.14 vs. 2.95 +/- 1.62 adducts/10(8) nucleotides; P = .085). Significant district and seasonal differences were found in subgroups with GSTM1-negative genotype. DNA adduct levels in placentas of the GSTM1-negative subgroup were higher in mothers living in the polluted district of Teplice than in Prachatice (P = .012). The adduct levels in placentas sampled in the summer period were higher than in the winter period in the GSTM1-negative population (P = .006). No effect of the NAT2 genotype on DNA adduct levels was observed.

Teplice program--the impact of air pollution on human health.

The aim of the Teplice Program is to investigate and assess the impact of air pollution on the health of the population in the district of Teplice, Czech Republic. Characterization of the air pollutants demonstrated unusually high concentrations during winter inversions of fine particles dominated by acidic sulfates, genotoxic organic compounds, and toxic trace elements. The major source of airborne fine particles is the burning of coal for heating and power. Human exposure and biomarker studies demonstrated large seasonal variations in air pollution within the Teplice District and higher seasonal average pollution levels than the comparative district, Prachatice. Personal exposures to fine particles and organic carcinogens [e.g., polycyclic aromatic hydrocarbons (PAH)] were correlated with excretion of PAH metabolites in urine, several trace metals in blood, and DNA adducts in white blood cells. Respiratory and neurobehavioral studies of school children were conducted using questionnaires and clinical measures. A significantly higher prevalence of adverse respiratory symptoms and decreased lung function were found in the Teplice district than in Prachatice. The neurobehavioral studies indicated significantly higher teacher referrals for clinical assessment in Teplice, but the majority of objective performance measures did not differ. Reproductive studies were conducted in both males and females. A study of the effects of exposure on pregnancy and birth found an excess prevalence of low birth weight and premature births in Teplice; these adverse effects were more common in infants conceived in the winter and whose mothers were smokers. Based on questionnaires and medical examination, the reproductive development of young men was not different between districts and seasons, however, measures of semen quality suggest that exposure to high levels of air pollution are associated with transient decrements in semen quality.

Isolation and characterization of glycocalyx in Streptomyces aureofaciens.

The surface layer, considered to be glycocalyx according to electron-microscopic observations, was separated from a low-production strain of Streptomyces aureofaciens by solubilization with urea and subsequent sonication. The isolation procedure was developed using various agents; neutral phosphatase served as a marker indicating the amount of the material released. The peripheral structure consisted predominantly of glycoprotein and differed from S-layers.

Cell volume maintenance and regulation in kidney tissues of various vertebrates.

Ouabain-sensitive and ouabain-insensitive mechanisms are shown to participate to a different extent in the cell volume maintenance in kidney tissues of various vertebrates. The ouabain-insensitive mechanisms predominate in the lamprey and in fresh-water fishes, whereas in the flounder, representing sea fishes, there is a complete predominance of the ouabain-sensitive mechanism. In the rabbit kidney cortex tissue the two systems play a comparable role. Analogous inter-species differences are displayed when the cells regulate their volume in anisoosmolar media.

Cell water content in carp kidney tissue slices as influenced by various osmotic agents.

Determination of water content in the surviving kidney tissue of the carp (Cyprinus carpio) is a convenient method for studying epithelial cell volume regulation under the influence of various osmotic agents and inhibitors. Artificial media of abnormal tonicities were used both as a purely experimental tool and as an attempt to model the behaviour of the tissue under extreme (physiological or pathological) conditions. In a medium hypertonic with mannitol the inhibition of the sodium-potassium pump by ouabain does not change the water content of the tissue (the same as in normal Krebs-Henseleit saline). On the other hand, in media which are hypotonic or hypertonic by addition of NaCl, KCl, Tris chloride or sodium benzenesulphonate the same inhibition invariably results in an increased water content. Thus, in contrast to the situation in normal or mannitol-hypertonic media, the ouabain-sensitive sodium-potassium pump does regulate the cell volume in artificial salines which are hypotonic or hypertonic by addition of electrolytes. 2,4-dinitrophenol often brings about a more pronounced swelling of the tissue than ouabain, suggesting the presence of an additional cell-volume regulating mechanism, ATP-dependent but ouabain-insensitive. The additional swelling is not always accompanied by a gain of sodium; hence it may not be correct to call the above mechanism a "second sodium pump". On the whole, the carp kidney tissue appears to behave as an osmometer with a certain incompressible volume, gaining or losing either sodium or potassium chloride in various abnormal media or with the two above inhibitors.

Diabetic gastroparesis: treatment with domperidone--a double-blind, placebo-controlled trial.

6 patients with severe symptomatic diabetic gastroparesis were studied in a placebo-controlled randomized manner to establish if intravenous domperidone accelerates the delayed gastric emptying of a semisolid homogenized meal. Domperidone, 10 mg intravenously, shortened the gastric emptying half-time in all of the 6 patients significantly, but not in the healthy control subjects. These results suggest that intravenous domperidone may be potentially beneficial to diabetic gastroparesis patients by improving delayed gastric emptying.

Ouabain-insensitive mechanism of cell volume maintenance in the carp kidney.

1) An ouabain-insensitive mechanism of cell volume maintenance, similar to that operating in the kidney tissue of mammals and in epithelial cells of the frog urinary bladder, has been demonstrated in the kidney tissue of carp (Cyprinus carpio)--there is no significant swelling even after 4-hour incubation with 10(-4) mol/l-1 ouabain. 2) The mechanism is not affected by 2-hour incubation with sodium in the saline replaced with tris or choline and, unlike in rabbit kidney cortex, cannot be inhibited by alkaline pH. 3) Neither a contractile mechanism sensitive to cytochalasin B, nor transport of divalent cations could be shown to be involved in this ouabain-insensitive cell volume maintenance in the carp kidney tissue.

Elective vital staining of mouse tumors with labelled dyes.

It was shown by vital staining in previous experiments (1) that carcinoma and sarcoma of mice could be selectively stained by certain dyes of the triphenyl methane sulphonic acid group. These findings were now confirmed in the present experiments by using labelled (125I) dyes of the same chemical group. In some mice the tumor tissue was the most radioactive, compared with other tissues. In other mice the tumor tissue was exceeded only by tissues which had a blood supply 5-time to 35-times higher. These were, in addition, excretory organs the excreta of which was highly radioactive (kidney and liver). The urine was 50-times more radioactive than the tumor and the bile was most probably not less. After deduction of the radioactivity of these two components, blood and excreta, from the radioactivity of the parenchyma tissue of the kidney and liver, the tumor tissue proved to be the most radioactive of all tissues tested. This indicates that tumor tissue retained the administered labelled dyes in higher concentration than all other tissues.

Limited usefulness of the breath test in evaluation of drug metabolism: a study in human oral contraceptive users treated with dimethylaminoantipyrine and diazepam.

Demethylation of 14C-dimethyl-N-aminoantipyrine (aminopyrine) and 14C-diazepam was measured by means of a breath test in women taking oral contraceptive steroids (OCS) and in controls not receiving OCS. Short-term half-life of 14CO2 in the breath after intake of 2 muCi of aminopyrine was significantly prolonged in women taking OCS when compared with controls. After intake of 2 muCi diazepam there were no statistically significant differences between the two groups. With 2 muCi diazepam or 10 muCi aminopyrine a biexponential decline of 14CO2 content in the breath was superimposed on a circadian, rhythm. 14CO2 in the aminopyrine breath test in the morning increased after the patients had risen from bed, whereas in the diazepam breath test the 14CO2 content decreased. It is concluded that the effect of OCS on drug metabolism is very specific. Furthermore, the appearance of 14CO2 in the breath does not depend only on hepatic microsomal demethylation.

PIP: 30 healthy female volunteers were tested in 5 groups of 6 persons each to evaluate whether oral contraceptives (OCs) had inhibiting effects on the disposal, via the hepatic microsomal enzyme system, of radiolabeled diazepam. Therefore, demethylation of radiolabeled aminopyrine and diazepam was measured by a breath test in women using OCs and in controls. Short-term half-life of radiolabeled carbon dioxide in the breath after ingestion of 2 mcCi of aminopyrine was significantly prolonged in women using OCs compared with controls. After ingestion of 2 mcCi of diazepan there was no statistically significant difference between the 2 groups. With 2 mcC; of diazepam or 10 mcCi of aminopyrine, a biexponential decline of radiolabeled carbon dioxide content in the breath was superimposed on a circadian rhythm, and the carbon dioxide in the aminopyrine breath test increased after patients had risen from bed whereas it decreased in the diazepam test. It is concluded that the effect of OCs on drug metabolism is very specific, and that furthermore, the appearance of radiolabeled carbon dioxide in the breath does not depend only on hepatic microsomal demethylation.