Alcohol consumption causes renal injury and compromises kidney function. The underlying mechanism of the alcoholic kidney disease remains largely unknown. In the present study, an alcoholic renal fibrosis animal model was first employed which mice received liquid diet containing alcohol for 4 to 12 weeks. The Masson’s Trichrome staining analysis showed that kidney fibrosis increased at week 8 and 12 in the animal model that was further confirmed by albumin assay, Western blot, immunostaining and real-time PCR of fibrotic indexes (collagen I and α-SMA). In vitro analysis also confirmed that alcohol significantly induced fibrotic response (collagen I and α-SMA) in HK2 tubular epithelial cells. Importantly, both in vivo and in vitro studies showed alcohol treatments decreased Smad7 and activated Smad3. We further determined how the alcohol affected the balance of Smad7 (inhibitory Smad) and Smad3 (regulatory Smad). Genome-wide methylation sequencing showed an increased DNA methylation of many genes and bisulfite sequencing analysis showed an increased DNA methylation of Smad7 after alcohol ingestion. We also found DNA methylation of Smad7 was mediated by DNMT1 in ethyl alcohol (EtOH)-treated HK2 cells. Knockdown of Nox2 or Nox4 decreased DNMT1 and rebalanced Smad7/Smad3 axis, and thereby relieved EtOH-induced fibrotic response. The inhibition of reactive oxygen species by the intraperitoneal injection of apocynin attenuated renal fibrosis and restored renal function in the alcoholic mice. Collectively, we established novel in vivo and in vitro alcoholic kidney fibrosis models and found that alcohol induces renal fibrosis by activating oxidative stress-induced DNA methylation of Smad7. Suppression of Nox-mediated oxidative stress may be a potential therapy for long-term alcohol abuse-induced kidney fibrosis.

In the present study, the activity of ACE (angiotensin-converting enzyme) in the aorta of senescent rats and rats treated with the NOS (NO synthase) inhibitor L -NAME ( N G -nitro- L -arginine methyl ester) or dexamethasone and the effect of low doses of ethanol (0.2–1.2 g/kg of body weight, daily for 8–12 days) on this activity were studied. We found that ACE activity increased with age and in response to L -NAME and dexamethasone treatment. Ethanol at a dose of 0.4 g/kg of body weight per day decreased ACE activity in the aorta of aged rats and of rats treated with L -NAME or dexamethasone to the level of activity in young control rats. The optimal ethanol dose (the dose inducing a maximum decrease in ACE activity) increased with increasing doses of dexamethasone: 0.4 g/kg of body weight per day at 30 μg of dexamethasone/kg of body weight and 0.8 g/kg of body weight per day at 100 μg of dexamethasone/kg of body weight. It was also found that optimal doses of ethanol increased the number of cells in the thymus of rats treated with dexamethasone. The optimal dose of ethanol of 0.4 g/kg of body weight per day, which induced a maximum decrease in ACE activity in rat aorta, corresponded to a dose of 30 g of ethanol/day, which, according to epidemiological data, produces a maximum decrease in the incidence of cardiovascular disease in humans. In conclusion, the decrease in ACE activity in vessels may be one of the main mechanisms of the beneficial effects of low doses of ethanol on human health.

Chronic alcohol muscle disease is characterized by reduced skeletal muscle mass precipitated by acute reduction in protein synthesis. The pathogenic mechanisms remain obscure, but several lines of evidence suggest that increased oxidative stress occurs in muscle in response to alcohol and this may be associated with impaired α-tocopherol status. Potentially, this implies a therapeutic role for α-tocopherol, especially as we have shown that supplemental α-tocopherol may increase the rate of protein synthesis in normal rats [Reilly, Patel, Peters and Preedy (2000) J. Nutr. 130 , 3045–3049]. We investigated the therapeutic effect of α-tocopherol on plantaris muscle protein synthesis in rats treated either acutely, chronically or chronically+acutely with ethanol. Protein synthesis rates were measured with a flooding dose of L -[4- 3 H]phenylalanine. Protein, RNA and DNA contents were determined by standard laboratory methods. Ethanol caused defined metabolic changes in muscle, including decreased protein, RNA and DNA contents in chronically treated rats. In acute or chronic+acute studies, ethanol suppressed fractional rates of protein synthesis. α-Tocopherol supplementation did not ameliorate the effects of either acute, chronic or chronic+acute alcohol on plantaris muscle protein content or rates of protein synthesis. In control animals (not treated with alcohol), α-tocopherol supplementation decreased muscle protein content owing to increases in protein turnover (both synthesis and degradation). α-Tocopherol supplementation is not protective against the deleterious effects of alcohol on protein metabolism in skeletal muscle.

We investigated the effects of smoking cessation or alcohol restriction on metabolic and fibrinolytic variables in Japanese men. In the smoking study, 35 male subjects [32±1 (S.E.M.) years] who habitually smoked cigarettes (29±3cigarettes/day) were told either to keep their usual smoking habits for 1week, or to abstain from cigarette smoking, using a randomized crossover design. In the alcohol study, 33 male subjects (37±1years) who habitually drank alcohol (64±6ml of ethanol/day) were told either to keep their usual drinking habits for 3weeks, or to reduce alcohol intake by at least up to a half of their usual drinking amount, using a randomized crossover design. In each study, venous blood samples were drawn after a 12-h overnight fast on the last day of each period, and metabolic and fibrinolytic variables were measured. One-week smoking cessation significantly increased serum high-density lipoprotein (HDL) cholesterol levels ( P <0.05), and significantly decreased serum lipoprotein (a) levels ( P <0.01) and plasma plasminogen activator inhibitor-1 levels ( P <0.05). In contrast, 3-week alcohol restriction significantly decreased serum HDL cholesterol levels ( P <0.05) and plasma tissue plasminogen activator levels ( P <0.05). These results suggest that smoking cessation has substantial and immediate benefits on lipid and fibrinolytic variables in habitual smokers, whereas alcohol restriction increases cardiovascular risks, in some respects, in habitual drinkers.

1. This cross-sectional study examined the influence of alcohol intake on organ injuries in normotensive and hypertensive subjects. 2. A total of 514 normotensive subjects and 302 never-treated hypertensive subjects were screened from 4557 men who entered the health check programme of our institute during the period 1990 to 1994. According to the daily alcohol consumption data reported by a self-administered questionnaire, the normotensive and hypertensive subjects were both classified into four categories; very light and non- (0–10 ml of ethanol), light (11–29 ml), moderate (30–58 ml) and heavy (≥59 ml) drinker groups. In these four pairs of groups, organ injuries in the heart, kidney and optic fundus were evaluated and serum lipids were measured. 3. Although the blood pressure levels were similar among the four groups of hypertensive subjects, the electrocardiographic findings of left ventricular hypertrophy were significantly more common among the moderate and heavy drinkers but not in the light drinkers compared with the very light and non-drinkers (very light and non-drinkers 25%, light drinkers 23%, moderate drinkers 38%, heavy drinkers 40%; P = 0.026). The alcohol intake increased the serum level of high-density lipoprotein cholesterol in a dose-dependent manner without changing the total cholesterol level; however, the serum γ-glutamyl transpeptidase and triacylglycerol levels were increased in the moderate and heavy drinkers. Urinary albumin excretion and fundoscopic lesions were not associated with the drinking habit in either the normotensive or hypertensive subjects. 4. These data suggest that habitual alcohol consumption exceeding 29 ml per day facilitates the development of left ventricular hypertrophy in hypertensive patients. Among the hypertensive subjects, light drinkers consuming 11 to 29 ml of ethanol daily showed preferable profiles in terms of organ injuries and risks of cardiovascular diseases.

1. In the present study we have examined the expression of pancreatitis-associated protein mRNA in mouse pancreas and small intestine and determined the effect of a number of factors on the steady-state level of the RNA. 2. The normally low level of pancreatitis-associated protein mRNA in pancreas increased severalfold after 6 h of hypoxia, reaching peak levels (approximately 10-fold greater than normal) after 24 h hypoxia. After 3 days' hypoxia pancreatitis-associated protein mRNA levels were again undetectable. 3. In the pancreas the level of pancreatitis-associated protein mRNA was also increased by alcohol and iron overload, but not by paracetamol. 4. In the small intestine expression of pancreatitis-associated protein mRNA was higher in normal ileum than in duodenum. In the ileum pancreatitis-associated protein mRNA levels were increased 7 to 15-fold after 6 h hypoxia, reaching peak levels by 24 h. Levels declined after 3 days' hypoxia, but remained higher than normal. 5. In the ileum long-term (4 weeks) dietary iron deficiency reduced pancreatitis-associated protein mRNA levels compared with control fed mice, whereas parenteral iron overload increased pancreatitis-associated protein mRNA levels. 6. The data presented suggest regulation of pancreatitis-associated protein gene expression by both oxygen tension and iron status.

1. The acute effects of a moderate dose of ethanol (1 g/kg body weight) on heart rate and blood pressure variability and baroreflex sensitivity were studied in 12 healthy male subjects in a juice-controlled experiment. Electrocardiographic and finger blood pressure data were recorded and stored in a minicomputer during 5 min of controlled breathing (15 cycles/min) and during deep breathing (5 s inpiration, 5 s expiration, four cycles) before drinking and hourly thereafter for 3 h. 2. Mean breath alcohol concentration rose to 18.9 mg/100 ml. In the time domain analysis, the root mean square difference of successive R-R interval decreased significantly with ethanol as compared with the juice experiment. The difference remained statistically significant even after adjustment for the shorter R-R interval after alcohol. In the frequency domain analysis the high-frequency (0.15-0.5 Hz) spectral power showed a significant decrease after alcohol intake. Also, the index of sensitivity of the baro-receptor reflex (square root of R-R interval power/systolic blood pressure power) decreased significantly in the high-frequency component. Ethanol did not change finger systolic blood pressure, and power spectral analysis did not show significant variability in blood pressure. 3. These data indicate that acute intake of moderate amounts of alcohol causes a significant decrease in heart rate variability owing to diminished vagal modulation of the heart rate.

1. The cardiovascular effects of oral alcohol (0.5 g/kg body weight diluted to 300 ml in sugar-free orange juice) were compared with those of placebo in 10 normal subjects. Measurements were made while the subjects were supine and horizontal for 45 min and after 10 min of 45° head-up tilt. 2. After alcohol, plasma alcohol levels rose from 1.9 ± 1.3 to 61.6 ± 6.5 mg/100 ml. After placebo, plasma alcohol levels did not increase. After alcohol and placebo, supine blood pressure was unchanged; heart rate, both supine and during tilt, rose after alcohol only. 3. After alcohol, superior mesenteric artery and digital skin blood flow increased and calculated vascular resistances fell. There was no change after placebo. 4. Forearm blood flow, forearm vascular resistance and cardiac index did not change in either phase, except for a fall in cardiac index during tilt but only after alcohol. 5. In conclusion, the acute ingestion of 0.5 g of alcohol/kg body weight in normal subjects raised heart rate and actively dilated the superior mesenteric artery and digital skin vessels. There was no effect on blood pressure, cardiac output and skeletal muscle vascular tone. During head-up tilt after alcohol, there was a tendency for blood pressure to fall with a compensatory rise in heart rate.

1. Electron paramagnetic resonance spectroscopy was used to study free-radical signals in freeze-clamped frozen liver tissue from rats after a 1 year period of dietary supplementation with alcohol, iron, or alcohol and iron. In alcohol-fed, iron-fed and alcohol- and iron-fed animals, mild histological damage was seen on light microscopy and evidence of mitochondrial and nuclear injury was identified by electron microscopy. 2. Subcellular fractionation studies showed an increase in the activity of the peroxisomal marker catalase ( P <0.01) in alcohol-fed rats compared with controls, but a fall of 82% ( P <0.001) in alcohol- and iron-fed animals. The activity of the mitochondrial marker succinate dehydrogenase rose by 7% (not significant) in alcohol-fed animals and by 17% (not significant) in iron-fed animals, but fell by 94% ( P <0.001) in alcohol- and iron-fed animals, suggesting serious impairment of mitochondrial function. 3. Iron overload was substantial in the iron-fed animals and there was an excellent correlation between liver iron concentration and iron-derived signals by electron paramagnetic resonance spectroscopy ( P <0.001). A clear free-radical signal of g = 2.003–2.005 was detected in all liver samples, but there was no significant difference in the magnitude of this signal in any study group. 4. The absence of any increase in the stable free-radical signal, even in the presence of considerable hepatic damage, does not support the hypothesis that free radicals mediate alcoholic liver disease in this animal model, although the results cannot be taken as proof against this hypothesis.

1. The short-term (120 min) kinetics of Zn turnover has been studied in control subjects and patients with alcoholic liver disease after intravenous injection of 0.5 mg of 96.5% enriched 70 ZnCl 2 . 2. The 70 Zn enrichment of plasma was found closely to obey two-compartment kinetics and the derived two-component decay equation has been used to calculate the size and turnover of the initial two rapidly exchanging pools of body Zn. 3. In normal subjects isotopic Zn appears initially to equilibrate with the whole of the plasma Zn which comprises the first metabolic compartment, pool a. This has a size of 0.72 ± 0.1 μmol/kg. 70 Zn equilibration then occurs with a second compartment, pool b, consistent with a rapidly exchanging liver Zn pool of size 3.60 ± 0.93 μmol/kg. The fractional turnover rate of pool b was found to be fivefold slower than that of pool a. 4. In the alcoholic group an expansion of pool a was observed (1.63 ± 0.39 μmol/kg), but the size of the second pool was not significantly different from that of control subjects (5.55 ± 1.0 μmol/kg), although its fractional turnover was significantly increased ( K ab : control subjects, 0.018 ± 0.002 min −1 , alcoholic patients, 0.031 ± 0.006 min −1 ). 5. These data therefore demonstrate that kinetic studies using stable isotopes of Zn can provide novel information on exchangeable Zn pools in man, but provide no support for the possibility of an underlying Zn depletion in patients with alcoholic liver disease.

1. This study was designed to determine prospectively whether changing alcohol consumption influenced the proportion of plasma linoleic acid independently of diet or smoking habits, and to evaluate changes in the plasma linoleic acid concentration as a potential marker of changes in alcohol consumption. 2. Fasting plasma fatty acid profiles were investigated in 72 male drinkers who were randomly assigned to drink low-alcohol beer or to maintain their usual drinking habits for a period of 4 weeks. 3. At entry to the study, a higher alcohol intake was associated with lower proportions of plasma linoleic acid, arachidonic acid and dihomolinolenic acid and higher proportions of plasma palmitoleic acid, independently of changes in body mass index. Smoking habits were unchanged and there were no major changes in diet during the period of the intervention. 4. Because the plasma palmitoleic acid concentration has been suggested as a possible marker of ‘at risk’ drinking, we investigated plasma fatty acid concentrations as indicators of alcohol intake. The plasma palmitoleic acid concentration was not a useful discriminator. Indices determined using logistic regression and combining plasma apolipoprotein A-II and linoleic acid concentrations gave better discrimination than either variable alone.

1. The effects of a single moderate dose of alcohol on blood pressure, heart rate and associated metabolic and endocrine changes were studied in 10 healthy subjects and compared with those of an isocaloric glucose control drink. 2. Systolic blood pressure rose at 1 h after both alcohol and the control drink. Therefore this early change was not specifically due to alcohol ingestion. Subsequently, there was a tendency (not statistically significant) for supine and erect systolic blood pressure to be reduced up to 8 h after alcohol ingestion. There were no consistent late changes in blood pressure observed over 7 days after alcohol. 3. Alcohol caused a marked tachycardia in both supine and erect postures which persisted beyond the time of detectable blood alcohol levels. 4. Blood sugar rose by a similar amount after alcohol and the isocaloric glucose control drink, but peak plasma insulin levels were higher after the control drink. 5. Plasma sodium rose in keeping with alcohol induced water diuresis. No significant changes in plasma potassium or magnesium were seen after alcohol. 6. Compared with the control drink there was no evidence from measurements of circulating adrenaline, noradrenaline, Cortisol, aldosterone or renin of activation of the sympathoadrenal axis, adrenal cortex or renin–angiotensin systems after alcohol.

1. The effects of alcohol (0.9 g/kg) compared with placebo (400 ml of orange juice) on plasma noradrenaline and 3,4-dihydroxyphenylethylene glycol levels, and on erect and supine blood pressures and heart rates, were studied in eight normal male volunteers. 2. Alcohol caused a rise in noradrenaline levels that commenced approximately 30 min after drinking and lasted about 4h. In contrast, 3,4-dihydroxyphenylethylene glycol levels fell immediately after alcohol administration and remained low for at least 6h. Acute alcohol administration alters noradrenaline catabolism, and may have a dual effect of increasing noradrenaline release and decreasing noradrenaline clearance. 3. Alcohol caused a transient rise in erect and supine blood pressures that preceded the rise in plasma noradrenaline. Thereafter erect blood pressures fell compared with control. This fall was associated with a progressive rise in both supine and erect rates, and reached a maximum several hours after the maximum levels of blood alcohol. 4. The major effect of acute alcohol administration is to lower blood pressure and induce a reflex tachycardia. Changes in noradrenaline and 3,4-dihydroxyphenylethylene glycol levels did not readily explain changes in blood pressure or heart rate, suggesting that alcohol induced changes in noradrenaline metabolism occur largely independent of changes in blood pressure and heart rate.

1. The capacity for glycolysis in muscle biopsies obtained from long-term heavy alcohol drinking patients has been compared with tissue from control subjects by assay in vitro of the total activities of glycogen phosphorylase, phosphofructokinase and fructose 1,6-bisphosphatase, key regulatory enzymes in the anaerobic glycolytic pathway. 2. Biopsies from 13 of 22 patients had type II fibre atrophy, and the activities of all three enzymes were reduced in these biopsies, when expressed in terms of DNA content, the most striking reduction being in phosphofructokinase activity. 3. The amount of glycogen in the tissue correlated closely with these enzyme activities and was slightly lower in the most atrophic tissue, when expressed in terms of DNA content. 4. The activities of acid and neutral α-glucosidases were similar in biopsies from control subjects and patients with various severities of alcohol myopathy. 5. The reduced activities are consistent with a reduced proportion of type II fibre muscle mass in these patients, and suggest that there may be a reduced capacity for glycolysis with resultant reduced lactate production. Whether the changes in enzyme activities are primary to the selective atrophy remains to be established.

1. δ-Aminolaevulinate dehydratase activity was measured in liver biopsy specimens from 13 patients with porphyria cutanea tarda, 13 alcoholics and 10 control subjects. 2. The mean enzyme activities in liver of porphyria cutanea tarda and alcoholics were 34% ( P < 0.001) and 49% ( P < 0.001) respectively of the control level. 3. Heat treatment of liver supernatant caused slight inhibition of the enzyme activity in porphyria cutanea tarda and in controls. 4. Addition of Zn 2+ to liver supernatants slightly increased the enzyme activity in both porphyria cutanea tarda and controls. 5. The addition of liver supernatant from the porphyria cutanea tarda to purified bovine 6–aminolaevulinate dehydratase did not result in suppression of the enzyme activity. 6. The apparent K m value of δ-aminolaevulinate dehydratase was 4.3 × 10– 4 mol/l for porphyria cutanea tarda livers and 3.3 × 10– 4 mol/l for control livers. The difference between the two was not significant.

1. Sixteen 44-year-old males with chronic high alcohol intake were investigated. Seventeen 44-year-old males with low alcohol intake from the same population served as controls. 2. Plasma noradrenaline concentrations did not differ significantly between individuals with high and low alcohol intake, neither at rest nor after acute stimulation induced by ambulation for 15 min. However, 63% (10 out of 16) of the individuals with high intake showed resting values within the upper quartile range for individuals with low intake. 3. Plasma renin concentration was twice as high ( P < 0.01) in the group with high alcohol intake as in the group with low intake. 4. Systolic as well as diastolic blood pressure was significantly higher ( P < 0.01) in the group with high intake compared with the group with low intake. 5. Sympathetic nerve activity, as defined from measurements of plasma noradrenaline concentration, is not uniformly increased in individuals with chronic high alcohol intake. The mechanisms behind the increased plasma renin level as well as the possible role of the renin-angiotensin system in alcohol-induced hypertension remain unsettled.

1. Folate deficiency is commonly found in alcoholic subjects although the causative mechanism is uncertain. It has been suggested that microsomal enzyme induction resulting from chronic alcohol ingestion might accelerate the rate of folate catabolism thus causing deficiency. 2. By using an experimental animal model to determine the rate of catabolism of [ 3 H]pteroylglutamate (folic acid) by the quantitative estimation of the two urinary catabolites p -[ 3 H]aminobenzoylglutamate and [ 3 H]acetamidobenzoylglumate, we have measured both the rate of folate catabolism and the extent of microsomal-enzyme induction in mice after acute and chronic alcohol ingestion. 3. Despite significant evidence of enzyme induction in the chronic alcohol group, there was no difference in the rate of folate catabolism after acute or chronic alcohol ingestion when compared with that of the controls.

1. The cerebral depressant effect of 30 ml of aqueous ethanol diluted to 25% and administered orally to 16 volunteer subjects was compared with a control group of 15 volunteer subjects. 2. The two parallel forms of the Watson-Glaser Critical Thinking Appraisal tests were employed as a measure of cerebral function. 3. The control group showed a small but statistically insignificant improvement on retesting with Watson-Glaser test form ZM after preliminary administration of form YM. 4. The relationship between the blood alcohol time curve and the alcohol effect was analysed for each individual subject, each subject being used as his own control. 5. The main peak cerebral depressant effect was substantial and occurred on average 25.5 min before the attainment of the peak blood alcohol concentration. 6. There was no significant correlation between blood alcohol concentration and contemporaneous cerebral impairment ( r = −0.01). 7. There was a highly significant correlation ( r = 0.60) between the effect upon cerebral function and the gradient of the blood alcohol curve at that time.

1. The influence of ethanol administration on the metabolism of non-esterified fatty acids (NEFA) and on the utilization of blood-borne substrates and stored glycogen by leg muscles was examined in nine young healthy subjects at rest and during 40 min bicycle ergometer exercise at 50% of maximal capacity. Ethanol was administered by a constant-rate intravenous infusion (6 mmol/min), giving an arterial concentration within the range of 9·3–15·6 mmol/l. Turnover of NEFA and regional exchange were evaluated with [ 14 C]oleic acid, and muscle metabolites were analysed from needle biopsy samples. 2. Leg blood flow was not altered by ethanol. Alanine in arterial blood was reduced by approximately 25% after ethanol administration during exercise. A slight decrease in the concentration of NEFA in arterial plasma was observed after ethanol, associated with an increase in the fractional turnover of oleic acid, indicating an increased removal of NEFA from the plasma. The rate of uptake of NEFA in the leg was only slightly decreased. 3. The total glycogen depletion in leg muscle during exercise was not influenced by ethanol. The pattern of glycogen utilization in different muscle fibres was changed by ethanol and found to be dependent on the fibre composition of the respective subject. In the control experiments, but not after ethanol administration, a negative correlation was observed between glycogen utilization during exercise and the fraction of type I fibres. 4. It is concluded that ethanol administration during exercise (a) decreases leg uptake of NEFA due to a lower concentration of NEFA in plasma, although this lower uptake is compensated by an increased fractional oxidation, (b) does not influence total glycogen depletion in leg muscle, (c) changes the pattern of glycogen utilization in different muscle fibres, (d) influences only to a small extent the energy supply to working muscles, the most predominant isolated effect being on inhibition of the lactate release.

1. We have studied activity of δ-aminolaevulinate synthase in needle liver biopsy specimens obtained from 12 human cirrhotic subjects, five subjects who had ingested anticonvulsants and from control subjects. Liver iron concentrations and quantitative urinary excretions of porphyrins plus their precursors were also determined. 2. In liver homogenates from subjects of each group, addition of an exogenous system for generation of succinyl-coenzyme A (CoA), including succinic thiokinase, resulted in appreciable enhancement of activity beyond that obtained without this system. 3. Mean activities for δ-aminolaevulinate synthase were not significantly different among patient groups when assayed without the exogenous succinyl-CoA-generating system, but liver homogenates from cirrhotic patients and subjects ingesting anticonvulsants had significantly higher activities than control subjects in the presence of the succinyl-CoA-generating system. 4. Although mean liver iron concentration in the cirrhotic group was slightly higher than in control subjects, and in control subjects there was some correlation between liver iron concentration and activity of δ-aminolaevulinate synthase, variations in this activity could not be accounted for solely on the basis of chronic hepatic deposition. Nor were these variations ascribable to differences among subjects in ingestion of ethanol before biopsy or severity of hepatic inflammation as judged biochemically and histologically. 5. Cirrhotic subjects excreted more uro- and copro-porphyrin than control subjects, whereas subjects ingesting anticonvulsants excreted more δ-aminolaevulinic acid and porphobilinogen than control subjects. However, these increases were small and not sufficient to account for all the increased δ-aminolaevulinic acid which potentially could have been formed by these subjects. 6. These considerations raise the possibilities that, in vivo : (a) rate of human hepatic synthesis of δ-aminolaevulinic acid is modulated by the supply of succinyl-CoA; (b) the rate of hepatic synthesis of haem is increased in cirrhotic patients and subjects ingesting anticonvulsants; (c) other important routes exist for disposition of haem precursors in these subjects, besides utilization for haem synthesis.

1. Male death rates from hypertension and stroke in England and Wales in 1949–53 were highest in those socio-economic and occupational groups with the highest death rates for cirrhosis of the liver (and presumably with highest alcohol intake). 2. In prevalence data from the Busselton population in Western Australia in 1969, there was a significant association between hypertension and a history of heavy drinking. 3. Together with other data, these observations suggest that up to 30% of hypertension in affluent countries may prove to be attributable to the use of alcohol.

1. Hepatic alcohol dehydrogenase activity and leucocyte ascorbic acid content were measured in thirty-five patients with liver disease and in ten control subjects with duodenal ulcer. The patients with liver disease were divided into three groups consisting of non-drinkers, moderate drinkers and alcoholic/heavy drinkers. 2. There was no significant difference in hepatic alcohol dehydrogenase activity between the groups with liver disease, but all patients had less than half the hepatic alcohol dehydrogenase activity of the control subjects ( P < 0·001). 3. The ascorbic acid in leucocytes was significantly lower in the alcoholic/heavy drinker group than that in the control subjects ( P < 0·02) when the Student's t -test was applied, but no significant difference was found when the Mann—Whitney U -test was used. 4. A correlation coefficient of r = 0·77 ( P < 0·001) was observed among the thirty-five patients with liver disease when hepatic alcohol dehydrogenase activity was compared with leucocyte ascorbic acid content. An insignificant correlation ( r = 0·332) was found in the control subjects with no liver disease. 5. This comparison was also significant among non-drinkers with liver disease ( r = 0·873; P < 0001), moderate drinkers ( r = 0·739; P < 0·02) and alcoholic/heavy drinkers ( r = 0·702; P < 0·005). 6. The addition of ascorbic acid in vitro (0·5–10 mmol/l) had no effect on the activity of alcohol dehydrogenase. 7. The relation between hepatic alcohol dehydrogenase activity and leucocyte ascorbic acid content is probably a consequence of liver disease, as opposed to any specific effect of ascorbic acid deficiency or alcohol consumption on alcohol dehydrogenase activity.