Two new reactive targets of 2,5-hexanedione in vitro - beta-alanine and glycine.

In this study, we found that two amino acids reacted with 2,5-hexanedione to form new reaction products in vitro, respectively. In the reaction of beta-alanine and 2,5-hexanedione, a reaction product was obtained and analyses of obtained results showed it was 3-(2,5-dimethyl-1H-pyrrol-1-yl)propanoic acid; in the reaction of glycine and 2,5-hexanedione, a reaction product was also obtained and analyses showed it was (2,5-dimethyl-1H-pyrrol-1-yl)acetic acid. Two reaction products were found to be oxidized easily; in addition, the latter was more easily to be oxidized than the former in the air. Our discoveries demonstrated that reactions between amino acids and 2,5-hexanedione could exist possibly in vitro. At present, it is clear that 2,5-hexanedione causes either axon atrophy or swelling, but the underlying molecular mechanism is still unclear. Since both beta-alanine and glycine are considered as neurotransmitter in the central nervous system, the reaction products remain to be identified in vivo.

Fertilization rates of small-head sperm in conventional IVF and ICSI.

Five men produced 23 ejaculates in 23 assisted reproductive technology cycles for semen analysis. In 11 of the 14 ICSI cycles and in 5 of the 9 IVF cycles, small-head sperm were found in more than 70% of the cells, having a length of < 3.5 micro m and a width of < 2 micro m. In 6 of the 14 ICSI cycles, the embryologist who performed the ICSI was not alerted to e presence of small-head sperm. Subsequent fertilization rate was significantly lower than a) the fertilization rate of the remaining 5 ICSI cycles that acted as a control, in which the embryologist was alerted to the presence of small-head sperm, and b) was lower than the 3 ICSI cycles with normal sperm head (p < .05). The 8 cell embryo and blastocyst rates in the former group were also decreased, although not significantly, compared to the latter groups. Likewise, in the 5 IVF cycles with small-head sperm, the fertilization rate was significantly lower than in the 4 IVF with normal sperm head (p < .05). Embryologists should be alerted when a high percentage of small-head sperm are detected during routine semen analysis because they may be associated with reduced fertilization and embryo outcome.

Long-term treatment with sterigmatocystin, a fungus toxin, enhances the development of intestinal metaplasia of gastric mucosa in Helicobacter pylori-infected Mongolian gerbils.

BACKGROUND: Helicobacter pylori is a human gastric carcinogen. Sterigmatocystin (ST), a fungus toxin, is a risk factor of gastric cancer. Cytotoxin-vacuolation toxin A (VacA) present in supernatants of H. pylori suspensions can cause gastritis and ulcer. The aim of this study was to examine the effects of H. pylori, ST and VacA in Mongolian gerbils. METHODS: Male Mongolian gerbils (n = 196) were treated with H. pylori supernatants (10 ml/1000 mg) mixed with diet or inoculated intragastrically with H. pylori alone or with ST (100 or 1000 ppb), and then killed 27 months later. Gastric tissue sections were stained with haematoxylin and eosin (H&E), periodic acid-Schiff (PAS), Alcian blue (AB, pH 2.5) and with immunostaining for PCNA and p53 expression. RESULTS: In H. pylori-infected gerbils, the normal mucosa was replaced by hyperplastic epithelium. Severe gastritis, cystic dilatation of gastric glands, hyperplastic polyps and intestinal metaplasia were observed. In H. pylori + ST (1000 ppb) gerbils, intestinal metaplasia was significantly more frequent than in H. pylori alone animals. No pathological changes were observed in the H. pylori supernatant group. Osseous metaplasia was observed in the H. pylori + ST (100 ppb) group. Serum gastrin levels of the H. pylori + ST (1000 ppb) group were significantly higher than those of the other groups. PCNA labelling index and p53 index of infected gerbils were significantly higher than those of uninfected groups. CONCLUSION: H. pylori causes gastritis, ulcer and intestinal metaplasia. ST enhances the development of intestinal metaplasia and increases gastrin levels in H. pylori-infected Mongolian gerbils.

Long-term Treatment with Sterigmatocystin, a Fungus Toxin, Enhances the Development of Intestinal Metaplasia of Gastric Mucosa in Helicobacter pylori-infected Mongolian Gerbils.

BACKGROUND: Helicobacter pylori is a human gastric carcinogen. Sterigmatocystin (ST), a fungus toxin, is a risk factor of gastric cancer. Cytotoxin-vacuolation toxin A (VacA) present in supernatants of H. pylori suspensions can cause gastritis and ulcer. The aim of this study was to examine the effects of H. pylori, ST and VacA in Mongolian gerbils. METHODS: Male Mongolian gerbils (n = 196) were treated with H. pylori supernatants (10 ml/1000 mg) mixed with diet or inoculated intragastrically with H. pylori alone or with ST (100 or 1000 ppb), and then killed 27 months later. Gastric tissue sections were stained with haematoxylin and eosin (H&E), periodic acid-Schiff (PAS), Alcian blue (AB, pH 2.5) and with immunostaining for PCNA and p53 expression. RESULTS: In H. pylori-infected gerbils, the normal mucosa was replaced by hyperplastic epithelium. Severe gastritis, cystic dilatation of gastric glands, hyperplastic polyps and intestinal metaplasia were observed. In H. pylori + ST (1000 ppb) gerbils, intestinal metaplasia was significantly more frequent than in H. pylori alone animals. No pathological changes were observed in the H. pylori supernatant group. Osseous metaplasia was observed in the H. pylori + ST (100 ppb) group. Serum gastrin levels of the H. pylori + ST (1000 ppb) group were significantly higher than those of the other groups. PCNA labelling index and p53 index of infected gerbils were significantly higher than those of uninfected groups. CONCLUSION: H. pylori causes gastritis, ulcer and intestinal metaplasia. ST enhances the development of intestinal metaplasia and increases gastrin levels in H. pylori-infected Mongolian gerbils.

Stomach cancer-related mortality rate is higher in young Japanese women than in men.

This study compares stomach cancer-related mortality rates in Japan with those in European and Asian countries and analyzes trends in stomach cancer-related mortality rates according to gender in young Japanese over the period of 1957-1997. From official death certification numbers and population estimates, we obtained stomach cancer-related mortality rate for all ages and various ages according to gender. Japan's ranking compared to other countries in death percentage of all cancers which are attributable to stomach cancer was fourth for both men and women. In Japan and Ireland, total elimination of deaths from stomach cancer in men resulted in increased life expectancy of 0.68 and 0.22 y respectively, whereas the corresponding figures for women were 0.42 and 0.14 y respectively. The sex ratios of stomach cancer-related mortality rates were 0.75, 0.63, 0.80 and 0.94 for 25-29, 30-34, 35-39 and 40-44 y age groups, respectively, in 1997. The sex ratio of relative risk ranged from 0.62 to 0.92 in 25-40 y age groups during the observation period. The life expectancy in 30-34 y age group increased by 0.66 y for men and 0.41 y for women in 1995 after elimination of stomach cancer-related deaths. Our results suggest that stomach cancer-related mortality rates are still high in Japan and young women are at higher risk of stomach cancer-related death relative to young men and that sex ratio is stable or slightly decreased over the 40-y period. It is important to monitor this trend continuously in the next few years.

Stomach cancer-related mortality.

In Japan stomach cancer remains the leading cause of cancer-related mortality. We analysed the annual mortality rate of stomach cancer in relation to age, gender and life expectancy in Japan between 1970 and 1995. The adjusted stomach cancer-related mortality rates decreased from 88.9 in 1970 to 45.4 per 100,000 in 1995 in males and from 46.5 to 18.5 per 100,000 in females. The male-female ratio for stomach cancer-related mortality in all ages was 1.9-2.5 during this 25-year period, and the mortality rate was higher in females than in males at young age. The negative contribution to life expectancy for stomach cancer in males was 0.65 years and 0.42 years in females, which is consistent with a higher mortality rate in males. This negative contribution was 41.8% of total cancer in 1970 and 39.4% in 1995 in males and 34.4% and 16.0%, respectively, in females. Our results demonstrated the need to take into consideration the characteristics of stomach cancer in young women and the effects of ageing when designing programmes aimed at prevention and control of this malignancy.

Absence of p53-mediated G1 arrest with induction of MDM2 in sterigmatocystin-treated cells.

P53 plays a critical role in G1 checkpoint after DNA damage. MDM2 gene is a p53 target gene and its protein forms a feedback loop with p53 and inhibits p53-mediated G1 arrest. Sterigmatocystin (ST) is a mycotoxin and carcinogen. In this study we show that exposure of cells to ST for 12 or 24 h resulted in failure of G1 arrest at both time points. Accordingly, p53 protein was not increased and p21WAF1 expression was inhibited at 12 h, and both proteins were weakly induced at 24 h after treatment with ST. Meanwhile, MDM2 protein was induced in a p53-dependent fashion by ST at both 12 and 24 h. The induction of MDM2 was coincident with the cellular responses of p53 and p21WAF1, and might contribute to the failure of G1 arrest in ST-treated cells. In addition, ST-treated cells exhibited G2M arrest, regardless of p53 status. Our results indicate that the carcinogenic effects of ST seem to be mediated by failure of p53-mediated G1 checkpoint.

Effect of distance and population size on patient trips in a prefecture of Japan: Application of a transportation distribution model to the demand for and supply of health services.

The gravity model, a method for analyzing transportation distribution in transportation engineering, was used to explain patient trips between ten health service regions in a Japanese prefecture. The OD (Origin-Destination) tables were constructed with zoning by regions, distinguishing between out- and inpatients. The observed trips were determined from the data of a survey conducted in 1992 that reported the locations of patient residences and chosen medical facilities.The base values used in the model calculations were the population size of each region and the road distance between the centers of regions. Problems of intrazonal trip were avoided by setting a mean intrazonal movement distance.This model was calibrated by the linear regression method with simultaneous validation by the index of correlation coefficients. The model was found to accurately simulate the effect of distance on the choice of medical facilities and the differences between the characteristics of in- and out-patients. The population value in use showed the relation not only with demand but also the supply of clinical services. It was suggested that the model presented here was useful in the allocation of medical resources and would help explain the relationship between suppliers and consumers of medical services.

Effects of methyl ethyl ketone, acetone, or toluene coadministration on 2,5-hexanedione concentration in the sciatic nerve, serum, and urine of rats.

OBJECTIVE: To clarify changes in the serum, nerve, and urinary levels of 2,5-hexanedione (2,5-HD) in rats on coadministration with methyl ethyl ketone (MEK), acetone (AC), and toluene (TO). METHOD: 2,5-HD alone or combined with MEK, AC, and TO was injected subcutaneously into a total of 306 male Wistar rats. The rats were divided as follows into 7 groups: (1) 2.6 mmol/kg 2,5-HD alone (HD) and (2) 2.6 mmol/ kg 2,5-HD combined with 2.6 mmol/kg MEK (HD + MEK), (3) with 2.6 mmol/kg AC (HD + AC), (4) with 2.6 mmol/kg TO (HD + TO), (5) with 13.0 mmol/kg MEK (HD + 5MEK), (6) with 13.0 mmol/kg AC (HD + 5AC), and (7) with 13.0 mmol/kg TO (HD + 5TO). 2,5-HD concentrations in the serum, sciatic nerve, and urine of rats were determined within 16 h of the injections and pharmacokinetic parameters were estimated. RESULTS: It was observed that (1) the 2,5-HD concentration and AUC value (area under concentration versus time curve) determined in the serum and nerve increased significantly in the cotreated groups as compared with the HD group; (2) the effect MEK had in elevating the 2,5-HD concentration and AUC in the serum and nerve was stronger than that of AC, and the effect AC had was stronger than that of TO; (3) a. dose increase from 2.6 to 13.0 mmol/kg for MEK and AC induced further increases in the 2,5-HD concentration and AUC determined in the serum and nerve; (4) elimination constants recorded for 2,5-HD (Ke) from the serum and nerve decreased in all the cotreated groups, and the degree of the decrease correlated inversely with the elevation in 2,5-HD concentration and AUC in the serum and nerve; and (5) urinary 2,5-HD concentrations measured in the 13.0-mmol/kg cotreated groups increased in parallel with the elevation in serum 2,5-HD concentrations. CONCLUSION: Coadministration of 2,5-HD with MEK, AC, or TO can increase the concentration and AUC of 2,5-HD in serum and the sciatic nerve, and these increases can be further enhanced by an increase in the concomitant doses of MEK and AC.

Relationship between 2,5-hexanedione concentrations in nerve, serum, and urine alone or under co-treatment with different doses of methyl ethyl ketone, acetone, and toluene.

To ascertain the relationship among 2,5-hexanedione (2,5-HD) concentrations in nerve, serum and urine, rats were injected subcutaneously with 2.6 mmol/kg 2,5-HD alone, or together with 2.6 or 13.0 mmol/kg of methyl ethyl ketone, acetone and toluene. 2,5-HD concentrations in sciatic nerve (NC), serum (SC) and urine (UC) were determined, and the linear regression between each two of NC, SC, and UC were calculated. There was good correlation between NC and SC, SC and UC in the 2,5-HD alone group, and good correlation between NC and SC in the co-treated groups. Co-treatment solvent had little effect on the relationship between SC and NC. 13.0 mmol/kg co-treated solvent tended to decrease the regression coefficients compared with 2.6 mmol/kg co-treated solvent. These results show that SC can be used in estimating NC in the 2,5-HD alone or co-treated groups, and UC can be used in estimating SC in the 2,5-HD alone group.

Long-term Effects of Various Iodine and Fluorine Doses on the Thyroid and Fluorosis in Mice.

OBJECTIVE: To elucidate the participation of the independent and combined long term effect of various concentrations of iodine and fluorine on the pathogenesis of goiter and fluorosis in mice. METHODS: Nine drinking water supplies with different iodine and fluorine content were prepared by combination of potassium iodate and sodium fluoride solutions in bidistilled water. The concentrations of iodide were: 1. iodine deficiency (ID): 0.0; 2. iodine normal (IN): 20.0; 3. iodine excess (IE) 2500.0 ug/l; and these of fluoride were: 1. fluoride deficiency (FD) 0.0; 2. fluoride normal 0.6; 3. fluoride excess (FE), 30.0 mg/l. A total of 288 Kunmim mice was divided into 9 groups consisting of 32 animals each and each group, in addition to basal diet, received one of following iodide/fluoride mixtures: ID+FD, ID+FN, ID+FE, IN+FD, IN+FN, IN+FE, IE+FD, IE+FN, IE+FE. By such manner, one half of the animals of each group was treated for 100 days and the other half for 150 days. RESULTS: It was found that ID only and IE only could both induce the goiter. FE induced dental fluorosis and increased fluorine content in the bone. In addition, fluorine also affected the thyroid changes induced by ID or IE. After 100 days of treatment, fluorine showed some stimulatory effect on the thyroid in ID conditions and inhibitory effect in IE conditions. After 150 days, however, the effects of fluorine on the thyroid reversed as compared with that of 100 days. On the other hand, difference of iodide intake could also increase the toxic effects of FE on the incisors and bones. The rate and degree of the incisor fluorosis, the fluorine contents in the bone were significantly higher in the ID+FE group than those in the IN+FE and IE+FE groups. CONCLUSIONS: Both iodine deficiency and excess induced goiter as well as other functional and histopathological changes in the mouse thyroid. Excessive fluorine caused fluorosis of incisors and limb bones. In addition, iodine and fluorine do have mutually interacting effects on both goiter and fluorosis in the experimental mice.

Evaluation of cutoff levels for screening of gastric cancer using serum pepsinogens and distributions of levels of serum pepsinogen I, II and of PG I/PG II ratios in a gastric cancer case-control study.

This study was carried out to determine the cutoff levels of serum pepsinogen (PG) I, II and their ratio of PG I/PG II for gastric cancer to establish a better screening system. Optimal cutoff levels for gastric cancer screening using serum pepsinogens were determined using Youden's index. The sensitivity, specificity and Youden's index for gastric cancer cases were calculated according to sex, age and the stage of gastric cancer, and the maximum Youden's index in each category was adopted as the cutoff level for gastric cancer screening using serum pepsinogens. The maximal Youden's index in all gastric cancer cases was 0.37, corresponding to a cutoff level of PG I < 40 (micrograms g/l) and PG I/PG II < 3.5. The sensitivity and specificity for gastric cancer cases of these cutoff levels were 0.50 and 0.87, respectively. In future, better criteria for gastric cancer screening have to be examined with the estimation of Youden's index in addition to other epidemiological methods such as ROC (receiver operating characteristic) curves and/or cost benefit analyses.

Different administration schedules of the same dose of 2,5-hexanedione influence the development of neuropathy and the toxicokinetics.

The same total dose (1.2 g/kg/week) of 2,5-hexanedione (2,5-HD) was administered subcutaneously at 100 mg/kg/12 hr, 200 mg/kg/24 hr, and 400 mg/kg/48 hr to three groups of Donryu rats. The peripheral neuropathy induced by 2,5-HD was confirmed by clinical observation every day, and neurophysiological measurements every 4 weeks. During the 15th week of this experiment, 2,5-HD concentrations in plasma 0.5 to 24 hours after injection were determined. It was found that the greater the dose of 2,5-HD per treatment injected, the earlier peripheral neuropathy developed. Toxicokinetic analysis showed that both the values of the area under the plasma concentration versus time curve and the half life of 2,5-HD were increased, but the excretion parameters (Ke) were decreased, in animals treated with 200 mg/kg/24 hr and 400 mg/kg/48 hr 2,5-HD.

Influence of different doses of methyl ethyl ketone on 2,5-hexanedione concentrations in the sciatic nerve, serum, and urine of rats.

Rats were injected subcutaneously with 2,5-hexanedione (2,5-HD 2.6 m mol/kg) alone (HD group) or with 2,5-HD and methyl ethyl ketone (MEK) (2.6 m mol/kg of each agent, HD&MEK group) or with 2,5-HD 2.6 m mol/kg and 5 times that dose (13.0 m mol/kg) of MEK (HD&5MEK group). The concentration of 2,5-HD in serum and in the sciatic nerve was determined 0.5, 1, 2, 4, 8, and 16 h after administration. Urinary 2,5-HD concentration was determined from the beginning of administration up to 16 h afterward. 1) The concentration of 2,5-HD in the serum, the sciatic nerve, and the urine was increased significantly (p < 0.05) in the co-administered groups; the higher the MEK doses were, the greater was the increase. 2) The clearance of 2,5-HD from both the serum and the sciatic nerve was delayed in the co-administered groups. The highest concentration in serum and the sciatic nerve appeared at 1 and 2 h respectively. After administration, the biological halflife (t1/2) of 2,5-HD from 1 to 8 h in serum was 6.5, 5.8 and 12.0 h for the HD, HD&MEK, and HD&5 MEK groups respectively. From 8 to 16 h, the t1/2 in serum was 1.2, 3.2 and 16.6 h for the HD, HD&MEK, and HD&5MEK groups, respectively. In nerve tissue, the prolongation of clearance in the co-administered groups was greater than that in serum, the t1/2 from 2 to 8 h being 5.2, 9.6 and 19.9 h for the HD, HD&MEK, and HD&5MEK groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[Seroepidemiological study on hepatitis C virus infection in an endemic area of hepatitis C virus].

To clarify the prevalence of hepatitis C virus infection in a rural area with high incidence of chronic liver disease in Japan, sera from 412 inhabitants, aged 20-89 years, collected in 1989-1990 and sera from 483 inhabitants in the same area, collected in 1982 were tested for anti-HCV (antibody to C100-3) with the first-generation enzyme-linked-immunosorbent-assay (ELISA). In addition, sera from 118 inhabitants, aged 20-49 years, collected in 1989-1990 were tested for HCV-RNA with the polymerase chain reaction technique, with use of primers from the 5'-untranslated region of the HCV genome. Anti-HCV was positive in 175 out of 412 sera collected in 1989-1990 (42.5%): prevalence was higher in male (54.0%) than in female (34.9%). The prevalence rates in the 20-29, 30-39, 40-49, 50-59, 60-69, and 70-89 year-old group were 0%, 14.3%, 51.9%, 41.7%, 49.1%, and 53.1%, respectively. On the other hand, in sera collected in 1982, the overall prevalence of anti-HCV was 42.5% (175 of 412). The prevalence rates in the 20-29, 30-39, 40-49, 50-59, 60-69, and 70-89 year-old group were 12.5%, 43.3%, 51.5%, 50.3%, 60.7%, and 68.9%, respectively. The prevalence rate of young adults less than 40 years old in 1982 were higher than that in 1989-1990. In short, the prevalence of hepatitis C virus infection in this area had altered a great deal. HCV-RAN was detected in 42 of 118 (35.6%).(ABSTRACT TRUNCATED AT 250 WORDS)

Correlation of serum antibody titers against hepatitis C virus core protein with clinical features by western blot (immunoblot) analysis using a recombinant vaccinia virus expression system.

In order to study the relationships among the clinical features of hepatitis C patients, the presence of hepatitis C virus (HCV) RNA in their blood, and their serum antibody titers against the core protein of virus and to study the antibody levels in asymptomatic HCV carriers, a recombinant vaccinia virus containing a core protein gene was constructed. The recombinant virus expressed a protein with a molecular mass of 22 kDa in RK-13 cells as determined by Western blot (immunoblot) analysis. By using the cell lysate of virus-infected cells and serially diluted serum samples, core antibody titers in the groups of patients in the chronic hepatitis phase and in the convalescent phase as well as in asymptomatic carriers were determined by enhanced chemiluminescence Western blot analysis. Almost all patients in the chronic phase were shown to have high antibody titers of more than 1:500,000 and with no exception had of HCV RNA in their sera. On the other hand, patients who had recovered naturally and were in the convalescent phase were shown to have significantly lower antibody titers, and the antibody was not detected in the lowest serum dilution of 1:500 in 43% of these patients (three of seven total patients). Antibody levels of patients who showed a good response to interferon treatment decreased to intermediate levels between those of patients in the chronic phase and those of patients in convalescent phase. The antibody titers in asymptomatic carriers varied considerably from 1:500,000 to 1:500, and 41% (11 of 27 total individuals) of these carriers showed a high titer equivalent to that of those in the chronic phase. Core antibody was detected consistently in the individuals in whom HCV RNA was detected. This system for core antibody might be useful for identifying the stage of an apparent HCV infection.

[Distribution of serum pepsinogen I, II values and their ratios in residents of a rural area].

In order to adopt pepsinogen levels as a screening indicator of gastric cancer, the serum pepsinogen I (PG I) and serum pepsinogen II (PG II) levels were studied using one thousand samples from a rural area and their ratios PG I/PG II were calculated. Samples were collected by stratified random sampling of the residents in S town with a population of about 10,000. The collected data was statistically analyzed to determine distributions by respective categories of sex and age. The following results were obtained. 1. The distributions of PG I, PG II and their ratio PG I/PG II levels largely deviated from normal distributions. Therefore, it is recommended that non-parametrical method is used to analyze this data. 2. The median of PG I level (49.3 micrograms/l) in males was statistically higher than that (43.6 micrograms/l) in females. On the other hand, a statistically significant difference between males and females was not observed in either the PG II value or the ratio PG I/PG II, respectively. 3. The pattern for changes in PG I values with age was different between males and females. While PG I levels were almost constant until the sixties, after the seventies levels showed a marked decrease in male subjects. In female subjects, an age effect in levels was not observed for PG I. On the other hand, PG II levels increased with age except for age group seventies, and the PG I/PG II ratios showed a decreasing tendency in both males and females.

Duodenal hemorrhage and dermal vasculitis associated with mixed connective tissue disease.

We describe a 23-year-old woman with mixed connective tissue disease (MCTD) who developed duodenal bleeding responsive to treatment with corticosteroid and recurrent skin eruptions due to leukocytoclastic vasculitis. The endoscopic findings revealed patchy, reddened, edematous, friable mucosa with scattered small ulcers and oozing of blood in the second portion of the duodenum. Endoscopic pictures from the onset to healing are presented. This appears to be the first recorded occurrence of such findings in MCTD.

[Ion chromatographic determination of inorganic anions in human serum].

The ion chromatographic method is a rapid and reliable technique for the simultaneous determination of inorganic anion concentrations in serum. We are investigating the effects of various diseases on the serum levels of the following inorganic anions: phosphate, bromide, nitrate, and sulphate. In this paper, we tested the intra- and intermeasurement reproducibility of the ion chromatographic method using pooled normal human serum. We found a good intrameasurement reproducibility for all of the above inorganic anions. The intermeasurement reproducibility was good for three of the inorganic anions but not for nitrate. We measured the serum levels of these inorganic anions in 241 inhabitants in one rural area. Using sera of normal inhabitants chosen from the above group, we used the mean value plus or minus two standard deviations as the normal range of each inorganic anion. Our values are in agreement with previously published reports. We found that the serum levels of nitrate and sulphate tended to be increased in inhabitants with liver or kidney disfunction respectively.