Nobuo Shimojo

Changes in amounts of cytochrome P450 isozymes and levels of catalytic activities in hepatic and renal microsomes of rats with streptozocin-induced diabetes

Hepatic microsomal cytochrome P450s, which are involved in the metabolism of drugs, hormones, prostaglandins and fatty acids, change when animals develop diabetes. We studied changes in cytochrome P450 isozymes in both hepatic and renal microsomes of rats with diabetes caused by streptozocin, and compared the results with changes in catalytic activities in the microsomes. In hepatic microsomes of diabetic rats, the amount of cytochrome P450 2E1, an acetone-inducible isozyme, was two and a half times that of control rats, and that of P450 4A2, a major renal isozyme, was three times that in the controls. The amounts of cytochrome P450s 2A1, 2C6, 2C7, 3A2 and 4A3 increased in hepatic microsomes of diabetic rats, and P450 2C11 decreased. Treatment with insulin restored these to the levels in the controls. The catalytic activities of aniline hydroxylation, 7-ethoxycoumarin O-dealkylation, testosterone 2 beta, 6 beta, 7 alpha, and 16 beta-hydroxylation, and omega-, (omega-1)-hydroxylation of lauric acid were high in the hepatic microsomes of diabetic rats, and testosterone 2 alpha and 16 alpha-hydroxylation activities were low. In renal microsomes of diabetic rats, cytochrome P450s 2E1, 4A2 and K-4 were induced, and omega- and (omega-1)-hydroxylation activities were high. These changes were reversed by insulin treatment. The induction and suppression of cytochrome P450 isozymes in diabetic rats were consistent with the changes in the catalytic activities. In both hepatic and renal microsomes, P450s 2E1 and 4A2 were induced, altered metabolism of ketones and fatty acids in diabetes may contribute to these changes.

Routine measurements of umbilical artery lactate levels the prediction of perinatal outcome

OBJECTIVE Our purpose was to compare lactate levels with acid-base balance in the umbilical artery with respect to the prediction of pregnancy outcome. STUDY DESIGN A prospective study of 4045 cord samples was performed. Lactate was measured with a new method that requires 5 microliters of blood and provides the result within 1 minute. RESULTS The umbilical artery lactate concentrations were significantly elevated in instrumental deliveries (2.65 +/- 1.2 mmol/L) and in emergency cesarean sections (2.44 +/- 1.7 mmol/L) compared with spontaneous vaginal delivery (1.87 +/- 0.94 mmol/L) (p < 0.001, p < 0.001). Lactate correlated significantly to fetal pH, hemoglobin, base deficit, PCO2, and HCO3-. Lactate was comparable to pH and base deficit in sensitivity, specificity, and positive and negative predictive values in relation to morbidity and mortality. CONCLUSION Umbilical artery lactate concentration and acid-base balance predicted perinatal outcomes with similar efficacies; however, its simplicity makes lactate analysis an interesting alternative in obstetric care.

Scalp blood lactate: a new test strip method for monitoring fetal wellbeing in labour

Objective To determine fetal scalp blood lactate with a new test strip method in parturients with normal and abnormal cardiotocograms during labour and to describe the relation to maternal lactate, fetal scalp blood pH, cord artery lactate and acid‐base balance.

Induction of renal cytochrome P-450 in hepatic microsomes of diabetic rats

We purified two forms of cytochrome P-450 which was induced in hepatic microsomes of diabetic male rates treated with streptozotocin. One of these corresponded to P-450j. The other form, designated P450 DM-2, had a minimum molecular weight 53000 and a CO-reduced absorption maximum at 452 nm. The P450 DM-2 efficiently catalyzed the omega- and (omega-1)-hydroxylation of lauric acid, but was not efficient in metabolizing aminopyrine, 7-ethoxycoumarin, aniline, N-nitrosodimethylamine, or testosterone. The NH2-terminal sequence of P450 DM-2 was identical to that of P450 K-5, the major renal cytochrome P-450. Both forms gave very similar electrophoretic patterns of proteolytic digests. P450 DM-2 and P450 K-5 are closely related forms.

Lactate in fetal scalp blood and umbilical artery blood measured during normal labor with a test strip method

A new test strip method was used to determine the lactate concentration in fetal scalp blood during normal labor. Sixty‐six fetal scalp blood samples were collected at cervical dilatations between 4 cm and 10 cm. The mean lactate value was 1.7 + 0.8 mmol/1 simultaneously (1S.D.) and the mean pH sampled simultaneously was 7.36 ±0.04 (1S.D.). The corresponding values for base deficit was 2.1 (±1.9) and for pC02 5.8 (±0.8). No difference was seen in lactate concentrations or pH values in early compared to late first stage of labor. The mean lactate concentration in the umbilical artery immediately after delivery was 3.7 mmol/l±1.2(lS.D.). The method was easy to handle and gave the result within 60 seconds.

Comparison of n-acetyl-β-d-glucosaminidase and alanine aminopeptidase activities for evaluation of microangiopathy in diabetes mellitus

The activities of urinary N-acetyl-beta-D-glucosaminidase (NAG) and alanine aminopeptidase (AAP) were measured in 207 diabetic patients and 57 healthy controls, and the relationship of these enzymes to different stages of diabetic microangiopathy was studied. Diabetics with clinical proteinuria had higher urinary NAG and AAP (17.7 +/- 1.9 and 42.8 +/- 4.9 U/g creatinine, mean +/- SE, respectively) than healthy controls (1.8 +/- 0.1 and 10.0 +/- 0.4) or diabetics without proteinuria. Among diabetics without proteinuria, NAG excretion in those with retinopathy was slightly higher than in those without (6.4 +/- 0.5 v 5.4 +/- 0.4), and AAP in those with retinopathy was significantly higher than in those without (23.0 +/- 1.5 v 17.4 +/- 0.8, P less than 0.01). Urinary albumin measured by radioimmunoassay and lysozyme in diabetics with retinopathy but without proteinuria was higher than those without retinopathy (P less than 0.001 and P less than 0.01). The increase in albumin was the greatest in diabetics with long duration of the disease (greater than or equal to 8 years); however, NAG and AAP increased more significantly in those with high hemoglobin A1c than in patients with long duration.(ABSTRACT TRUNCATED AT 250 WORDS)

Lactate in cord blood and its relationship to pH and catecholamines in spontaneous vaginal deliveries

The interrelationships between lactate and pH, nonadrenaline (NA), adrenaline (A) and dopamine (DA) were investigated in cord artery (CA) and vein (GV) blood at delivery. Sixty consecutive, spontaneous, vaginal deliveries with fetuses in cephalic presentation were assessed. Median gestational age at delivery was 40 weeks (range, 35-43). There were significant correlations between lactate and pH (P < 0.01), NA (P < 0.01), A (P < 0.05) and arterio-venous NA (P < 0.05) and DA differences (P < 0.01) in CA blood, while no variable correlated significantly to lactate in CV blood. The higher levels both of lactate and of catecholamines in CA blood are probably fetally derived. Dividing the material into high and low lactate subgroups (cut-off level, 75th percentile) showed a high lactate level to be associated with lower pH and higher catecholamine levels in CA blood, though the relationship was only statistically significant for pH. The levels both of catecholamines and of lactate were lower than those reported for cases of fetal distress, and reflect the lower level of fetal stress in the present series of normal deliveries. The low level of fetal stress and the differences in turnover rates between catecholamines and lactate might obscure their causal relationships, vis-a-vis fetal adaptation to extrauterine life during the course of parturition.

Fetal scalp and umbilical artery blood lactate measured with a new test strip method

Objective To compare the measurement of lactate in fetal scalp and umbilical artery blood by a new dry reagent strip method with a commercially available enzymatic method using plasma (Monotest).

Lactate and acid–base balance at delivery in relation to cardiotocography and T/QRS ratios in the second stage of labour

OBJECTIVE To compare foetal electrocardiogram (T/QRS ratio) and cardiotocography (CTG) during the second stage of labour with lactate and acid-base balance in cord artery blood at delivery. DESIGN Forty-six parturients delivered at the National University of Singapore were monitored during the second stage of labour with T/QRS ratios and CTG. At delivery blood from a segment of clamped cord was sampled for lactate and acid-base balance analyses. The Spearman Rank correlation, the Mann Whitney U-test and the Kruskal Wallis ANOVA were used when appropriate. RESULTS Maternal pushing time was significantly correlated to lactate (R = 0.51; P = 0.0003), pH (R = -0.38; P = 0.009) and base deficit (R = 0.33; P = 0.026), but not to T/QRS ratio (R = 0.002; P = 0.99). No significant correlation between lactate and T/QRS ratios (R = 0.06; P = 0.70) or type of CTG pattern was found (P = 0.10), though there were significant differences in pH (P = 0.029) and T/QRS ratios (P = 0.037) between groups with different FHR abnormalities. CONCLUSION Lactate increases progressively with maternal pushing time. No significant correlation was found between lactate and the T/QRS ratio. Lack of correlation is likely to be due to poor sensitivity of foetal ECG at this level of foetal stress, though the influence from transplacentally transferred maternal lactate cannot be excluded.

Regulation of renal cytochrome P450s by thyroid hormone in diabetic rats

Effects of thyroid hormone treatment on renal P450 expression in the diabetic rats were investigated. Diabetes produced by streptozotocin induced CYP4A2 and P450 K-2 (similar form with CYP2C23) but not P450 K-4 (similar form with CYP4A8) and induced lauric acid hydroxylation activity. The serum thyroid hormone level was decreased with diabetes. Treatment of diabetic rats with thyroid hormone (T3) as well as with insulin reversed the increase in the levels of CYP4A2 and P450 K-2. Thyroidectomy also induced CYP4A2 and P450 K-2 in the rat kidney. The increase was reversed by treatment of thyroidectomized rats with T3. These findings suggest that expression of CYP4A2 and P450 K-2 in rat kidney is suppressively regulated by thyroid hormone and the decrease in thyroid hormone level in the diabetic state affects the levels of CYP4A2 and P450 K-2.