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Acivicin is an inhibitor of γ-glutamyl transpeptidase and glutamine amidotransferase. When grown on a synthetic minimal agar medium, acivicin strongly inhibited the growth of Magnaporthe oryzae and Alternaria brassicicola, and to a lesser extent, Botrytis cinerea. However, only partial or marginal growth inhibition was observed with regard to Fusarium sporotrichioides and Fusarium graminearum. The growth retardation caused by acivicin was significantly alleviated by cultivating the fungus on a nutrient-rich medium. The inhibition of M. oryzae growth caused by 1 µmol l(-1) of acivicin on minimal agar medium was subdued by the addition of specific single amino acids, including His, a branched-chain amino acid (Leu, Ile or Val), an aromatic amino acid (Trp, Tyr or Phe), Met or Gln, at a concentration of 0·4 mmol l(-1). Trichothecene production by F. graminearum in trichothecene-inducing liquid medium was reduced significantly in the presence of acivicin despite its inability to inhibit growth in the trichothecene-inducing liquid medium. Foliar application of conidia in the presence of acivicin reduced the severity of rice blast disease caused by M. oryzae. These results suggest the usefulness of this modified amino acid natural product to mitigate agricultural problems caused by some phytopathogenic fungi. Significance and impact of the study: Fusarium head blight or scab disease and rice blast, caused by Fusarium graminearum and Magnaporthe oryzae, respectively, are major diseases of cereal crops that cause a significant loss of yield and deterioration in the quality of the grain. The present study investigated the effects of acivicin, a glutamine amino acid analog, on the physiology of various phytopathogenic fungi. Application of acivicin to a fungal culture and conidial suspension reduced mycotoxin production by the wheat scab fungus and the severity of rice blast, respectively. These results suggest the possibility that acivicin may serve as a lead compound to develop agricultural chemicals for the control of some plant diseases.

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BACKGROUND/AIM: It is well known that Helicobacter pylori changes its shapes according to various environmental conditions. The aim of this study was to examine the morphological differences between H. pylori in the duodenum and that in the stomach. METHODS: Duodenal biopsy specimens were obtained from patients with duodenal mucosal lesions, and these specimens were then histopathologically examined. Morphological and genetic properties of cultured H. pylori isolates were analyzed. RESULTS: H. pylori was identified in 16 out of 50 duodenal biopsy specimens. Along with the regular form, we found a fine form of H. pylori in the gastric metaplastic mucosa of ten duodenal specimens that was shorter in length and thinner in diameter than the regular helical form. There were no detectable differences between the ureA-ureB polymorphism of the duodenal fine form and that of gastric regular form in a single patient. As compared with patients without H. pylori in the duodenum, the prevalence of recurrent duodenal ulcers significantly increased in patients with the fine form (p < 0.05), but not in patients with the regular form. CONCLUSION: The fine form of H. pylori in the metaplastic duodenal mucosa could result from its adaptation to the duodenal environment and may be associated with the recurrence of a duodenal ulcer.

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This paper presents a backward movement control of an articulated vehicle via a model-based fuzzy control technique. A nonlinear dynamic model of the articulated vehicle is represented by a Takagi-Sugeno fuzzy model. The concept of parallel distributed compensation is employed to design a fuzzy controller from the Takagi-Sugeno fuzzy model of the articulated vehicle. Stability of the designed fuzzy control system is guaranteed via Lyapunov approach. The stability conditions are characterized in terms of linear matrix inequalities since the stability analysis is reduced to a problem of finding a common Lyapunov function for a set of Lyapunov inequalities. Simulation results and experimental results show that the designed fuzzy controller effectively achieves the backward movement control of the articulated vehicle.

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A 22-year-old Japanese man with a mediastinal germ cell tumor with mixed teratoma and hepatoid alpha-fetoprotein positive, and glandular yolk sac components, also demonstrated widely disseminated malignant histiocytosis involving the lungs, liver, pancreas, spleen, bone marrow and lymph nodes. Sensitivity to chemotherapy was acute and the patient died of disseminated intravascular coagulation syndrome within four months of his initial presentation.

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UNLABELLED: In order to examine the effect of dehydroepiandrosterone-sulfate (DHA-S) on the production of uterus-contractive substances in perinatal women, the concentrations of estrogen stimulated neurophysin (ESN), oxytocin, 13, 14-dihydro-15-keto-Prostaglandin F2 alpha(PGF2 alpha) and DHA-S related steroids in the maternal blood of 12 cases of primigravida were serially estimated after intravenous administration of 200mg DHA-S twice a week from 38 weeks of gestation till the onset of labor. RESULTS: 1) The duration of labor was significantly shortened in the DHA-S group (mean; 9.3 +/- 2.8 hours) compared to the 11 control cases (13.5 +/- 5.2 hours). 2) i) The ESN concentration, which correlated well with that of free estradiol-17 beta, increased significantly at 1 week after the Ist administration. ii) The oxytocin concentration, which correlated well with that of ESN, increased significantly at 39 weeks of gestation, the first stage of labor and especially at delivery compared to the control. iii) The PGF2 alpha concentration, which increased in parallel with those of DHA-S, estrone-S and pregnenolone-S, increased significantly at delivery in comparison with the control and correlated well with estrone-S in the DHA-S group. CONCLUSION: These findings suggest that DHA-S administration to perinatal women may enhance the production of ESN, oxytocin and PGF2 alpha in the fetoplacento-maternal compartment, which may result in the shortening of labor.

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Nineteen-hydroxyandrostenedione (19-OHA) is considered to be an obligatory intermediate of estrogen synthesis. To clarify the role of 19-OHA in the human ovary, the following experiments were undertaken. Ovarian and peripheral vein blood in mid follicular and luteal phase were obtained from 14 women during gynecological surgery. The concentrations of androstenedione (A), testosterone (T), 19-OHA, estrone (E1) and estradiol (E2) were measured by gas chromatography mass spectrometry using deuterium-labeled steroids as internal standard. The effect of human chorionic gonadotropin (hCG) on those steroids was also studied in mid luteal phase. The concentrations of 19-OHA in ovarian vein were 1.20 +/- 0.16 ng/ml (mean +/- SE) in mid follicular and 0.78 +/- 0.18 ng/ml in mid luteal phase. The steroid levels measured in ovarian vein were significantly higher than those in peripheral vein. Though the administration of hCG in mid luteal phase enhanced A and E2 levels in ovarian vein, the levels of T, 19-OHA and E1 were not altered. Significant correlation between the levels of E2 and A was observed while none of the steroids correlated with 19-OHA. These results suggest, for the first time, that 19-OHA is produced and secreted from the human ovary. The question as to the physiological and biosynthetic role of ovarian 19-OHA has to be answered in the future.

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The conversion of androgen to estrogen is mediated by aromatase activity, and 19-hydroxylase and C10-19lyase are seen to be involved in C19-demethylation. The present study is to demonstrate C10-19lyase activity in adrenal tissue. Fetal bovine and pig adrenal tissue homogenates were incubated with 14C-androstenedione(A) and NADPH under air. The products 14C-19-OHA, 14C-19-norA, with added carrier standard 3H-19-OHA, 3H-19-norA were separated and purified by T.L.C.. The identification of 19-norA was established by recrystallization and that of 19-OHA by C.C.D.. The 3H/14C ratios of the products 19-norA were found to be constant in repeated crystallization. The amount of 19-OHA from A in fetal adrenal tissue was 30.0nmol/h/g protein, and that of 19-norA from A was 12.59pmol/h/g protein. C10-19lyase activity from 19-OHA, calculated by the amounts of 19-norA in fetal, bovine and pig adrenal tissue, was 49.7, 3.50, 10.3pmol/h/g protein, respectively. This is the first report to demonstrate androgen C10-19lyase in human fetal adrenal tissue, and it is assumed that fetal adrenal tissue forms 19-norA via 19-OHA.

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19-Hydroxyandrostenedione (19-OHA) is secreted from the adrenal glands in men and women and also from the placenta during pregnancy. It has been found to cause hypertension in animal models. We have synthesized [7,7-2H2]-19-OHA with high deuterium content and, together with [7,7-2H2]A and [9,11-2H2]estrone (E1), have developed a quantitative assay of serum level 19-OHA, A, and E1 using the gas chromatography/mass spectrometry-mass fragmentography method to monitor individual subjects throughout pregnancy. The labeled 19-OHA, used as internal standard, showed only 6.73% of unlabeled compound. Recovery of standard 19-OHA, A, and E1 (5,000 pg each) added to male plasma was 97.4 +/- 2.3%, 96.3 +/- 2.1%, and 100.1 +/- 4.1% (mean +/- SD), respectively; the intraassay coefficient of variation was 2.1%, 3.5%, and 3.8%, respectively. Ten pregnant subjects without complications and 10 pregnant subjects near term with hypertension were selected (with informed consent). The 19-OHA and E1 serum concentrations of maternal venous blood from uncomplicated pregnancies increased significantly as gestation progressed (19-OHA: first trimester, 225 +/- 72; second trimester, 656 +/- 325; third trimester, 1,518 +/- 544 pg/ml), reaching the highest level at delivery (19-OHA: 1,735 +/- 684 pg/ml). Whereas a positive correlation was found between the level of 19-OHA and E1, no apparent change of the A level was observed during pregnancy. Levels of the three steroid hormones in pregnancy complicated by hypertension in the second and third trimester were not found to be significantly different from those of normal pregnancy (19-OHA of hypertensive subjects: second trimester, 762 +/- 349; third trimester, 1,473 +/- 491 pg/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

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To study the effects of dehydroepiandrosterone sulfate (DHA-S) on placental steroid metabolism and maternal steroidal profiles at term, the following in vivo and in vitro experiments were performed. Two hundred mg of DHA-S was given to five pregnant women 30 minutes prior to delivery. After delivery, the placenta was collected and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) and sulfatase activity was determined by measuring the rate of conversion of pregnenolone to progesterone and DHA-S to DHA. The amount of C21-delta 4-steroid in the placental tissue was measured by gas chromatography mass spectrometry (GC-MS) and compared with the control groups. The maternal serum concentration of several steroids was also measured by GC-MS before and after the administration of DHA-S. 3 beta-HSD activity in the placentae from the mothers who received DHA-S before delivery was significantly lower than in the controls. On the other hand, no significant change was observed in the activity of sulfatase. The serum concentration of progesterone (P) and 20 alpha-dihydro-P (20-P) before DHA-S loading decreased following the administration whereas estradiol (E), DHA, and androstenedione (A) levels increased. To study the direct effect of DHA-S and its related steroids on placental 3 beta-HSD activity, placental tissue samples were incubated with pregnenolone in vitro. Several other steroids were added simultaneously into the medium. It was observed that placental 3 beta-HSD activity was directly inhibited by DHA-S. These results indicate that DHA-S inhibits 3 beta-HSD activity in the placenta and subsequently causes a reduction in P and 20-P.

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19-Hydroxyandrostenedione (19-OHA), highly labeled with deuterium at position 7, was synthesized from unlabeled androstenediol diacetate. The deuterium labels were introduced into the 7-keto compound with dichloroaluminum deuteride to obtain [7-2H2]androstenediol. The labeled androstenediol diacetate was converted to the labeled 19-OHA by a five-step sequence without appreciable loss of deuterium. The labeled 19-OHA is useful as an internal standard for gas chromatography-mass spectroscopy analysis of the endogenous levels and as a tracer for in vivo metabolic studies.

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To study the serum levels of 19-hydroxyandrostenedione (19-OH-A), known as an obligatory intermediate of estrogen biosynthesis and considered to be one of the hypertensinogens, a method using GC-MS with application of synthesized [7,7-d2]androstenedione (A), [7,7-d2] 19-OH-A and [9,11-d2]estrone(E1) as internal standards was newly developed. Normal pregnant women and pregnant women complicated with hypertension near term were selected for the study. The levels of 19-OH-A and E1 increased significantly as gestation progressed [19-OH-A; 224.7 +/- 72.1 (1st trimester), 655.5 +/- 325.4 (2nd trimester). 1517.8 +/- 543.6 (3rd trimester)pg/ml], and a positive correlation was found between the levels of the two steroids. No apparent change was observed in A levels during the course of pregnancy. The mean levels of 19-OH-A in pregnancy complicated with hypertension at 2nd and 3rd trimester were 761.7 +/- 348.9 and 1473.0 +/- 491.4 pg/ml, which were compatible with those in normal pregnancy. The levels of 19-OH-A at delivery in maternal vein (MV) were 1735.1 +/- 683.9 pg/ml. Significantly higher levels of 19-OH-A were found in umbilical vein (UV) (1977.2 +/- 564.9 pg/ml) than those in umbilical artery (109.7 +/- 49.1 pg/ml). 19-OH-A concentration in term placental tissue was 16.3 ng/g.w.w. tissue. This is the first report to demonstrate the serum 19-OH-A levels measured by GC-MS and also to demonstrate the levels in the cord blood. The results indicate that 19-OH-A may be the product of pregnancy and may be derived from the feto-placental compartment.

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19-hydroxyandrostenedione (19-OH A), known for many years as an obligatory intermediate of estrogen biosynthesis, has recently been found to be an amplifier of aldosterone action and in itself a hypertensinogenic steroid (J. Steroid Biochem., 16; 329, 1982). To search the presence of a non-aromatizing andorgen 19-hydroxylase in fetal organs, metabolism of labeled androstenedione(A) in fetal adrenal, liver, lung, brain and kidney was studied. 19-hydroxylase activity was calculated by the amounts of 19-OH A formed from A. The tissue homogenate was incubated with [4-14C]A and NADPH for different periods of time under air. The product, [4-14C] 19-OH A with added carrier standard [6,7-3H] 19-OH A was separated and purified by TLC, acetylation and TLC to constant 3H/14C ratio. The identity of the product was established by C.C.D. and by GC-MS analysis of purified product 19-AcOA in a larger scale adrenal incubation of non-labeled A. 19-hydroxylase activity was 0.9 pmol/min/mg protein for adrenal, but there was a negligible quantity of 19-OH A in lung, brain and kidney. Estrogen production was also assayed by 3H-water method using [1 beta-3H, 4-14C] A (3H/14C = 69.9). No detectable amount of estrogen was found in incubation of any homogenate except for liver. These results indicate that the non-aromatizing 19-hydroxylase activity in fetal adrenal gland is much higher than that in other organs. This is the first report to demonstrate the presence of 19-hydroxylase in human fetal adrenal tissue. It is suggested that 19-OH A secreted from fetal adrenal may also be associated with serum concentration of fetal circulation.

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In order to study the changes of C21-steroid levels which included Pregnenolone (P5), 20 alpha dihydropregnenolone (20P5), 16 alpha hydroxypregnenolone (16P5), progesterone (P4) and 20 alpha dihydroprogesterone (20P4) in maternal peripheral blood during pregnancy and at delivery, these steroids were measured by GC-MASS with application of deuterated steroids as internal standard. The accuracy of GC-MASS method of these steroids was satisfactory with C.V. value of less than 6%. Total delta 5C21 steroid concentrations in course of pregnancy and at delivery were as follows; P5 (mean +/- S.D. ng/ml): 66.6 +/- 36.2 (1st trimester), 80.9 +/- 24.6 (2nd trimester), 147.7 +/- 30.1 (3rd trimester) and 299.7 +/- 178.3 ng/ml (at delivery), 20Ps: 212.6 +/- 102.5, 143.4 +/- 53.9, 248.9 +/- 58.8, 563.4 +/- 198.2 ng/ml, 16P5: 8.6 +/- 8.6, 8.1 +/- 5.2, 124.3 +/- 40.3, 378.5 +/- 180.0 ng/ml, respectively. P4 (43.0 +/- 28.0 ng/ml) and 20P4 (8.0 +/- 4.0 ng/ml) in 1st trimester showed gradual increase to maximum level (P4: 138.2 +/- 30.1 ng/ml, 20P4: 105.4 +/- 21.6 ng/ml) at pre-pain period, afterward decreased rapidly (P4: 70.9 +/- 23.2 ng/ml, 20P4: 59.8 +/- 19.3 ng/ml) at delivery. P5, 20P5 and 16P5 levels were found to be significantly higher in umbilical artery (UA) as well as in umbilical vein (UV) than those in maternal vein (MV) regardless of labor pain. P4 and 20P4 did not show any differences in MV regardless of labor pain. P4 in UV (pain+) and 20P4 in UA (pain-), however, showed significantly higher than P4 in UV (pain-) and 20P4 in UA (pain+). P5, 20P5, 16P5 and 20P4 levels were significantly lower in the case of anencephalic pregnancy (ANC) at 3rd trimester than in normal pregnancy, especially 16P5 levels (22.2 +/- 5.0 ng/ml) showed 1/5 of those in normal pregnancy. From the results obtained above, it is suggested that these delta 5C21 steroids are actively produced in the feto-placental unit in the course of pregnancy. The levels of these steroids reached maximum at delivery, but the levels of P4, 20P4 decreased toward delivery after maximum levels were shown in the stage of pre-labor pain. No significant difference of P4 level in the case of ANC suggested that P4 production correlated with placenta as well as maternal and fetal precursor. Decreasing of 20P4 and P4 level after the stage of pre-labor pain suggested that activity of 3 beta hydroxysteroid dehydrogenase was reflected by uterine contraction during labor.

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We reported on the unusually high isotope effect of non-aromatizing androgen 19-hydroxylase in sheep and dog adrenals and the validity of the [3H] water method using [19-3H3] androgen. We have extended the study to examine whether this 19-hydroxylation is catalyzed by a cytochrome P-450 dependent enzyme. Sheep adrenal homogenate (1.65 mg prot.) was incubated in the presence of NADPH (5.6mM) with [19-3H3, 4-14C]-androstenedione (A) (3.2 microM, 8.24 x 10(4) dpm 3H/micrograms, 3H/14C = 17.2) in a total of 1.2 ml PO4 buffer under air at pH 7.4 for 2, 5 and 10 min. [19-3H2, 4-14C]-19-hydroxy-A (19-OHA) with added carrier was purified through extraction, TLC, acetylation to form 19-AcOA, and further TLC to give 19-hydroxylase activity as assessed by the product isolation method. Simultaneously, the [3H] water was measured by distillation, and with correction by the apparent kinetic isotope effect (KH/KT = 11.8), used for assessment of 19-hydroxylase activity. The effects on the hydroxylation by cofactor (NADPH, NADH), incubation atmosphere (N2, CO/O2), cytochrome P-450 inhibitors (metyrapone, clotrimazole) and heating were measured by both methods. Compared to the complete system (89.6pmol/min/mg as 100%), carbon monoxide suppressed 15.8, 59.3 and 86.4% of the 19-hydroxylation when a CO/O2 ratio of 0.1, 1 and 9 was used, respectively. Replacement to nitrogen atmosphere decreased the activity by 93.8%. Replacement of NADPH with NADH (7.5mM) caused more than a 92.1% decrease in activity. Metyrapone at 50 and 200 microM and and clotrimazole at 2.5 and 10 microM suppressed the activity by 82.8, 90.4, 85.4 and 94.9%, respectively. A larger scale sheep adrenal incubation of A (250 microM) under 18O2 atmosphere and isolation of 19-AcOA were carried out in a similar manner. The gas chromatography-mass spectrometry analysis of the purified product showed 48.5% of the product to be 18O-labeled as [M+ + 2], m/e 346. Thus, the non-aromatizing androgen 19-hydroxylase requires NADPH and molecular oxygen. It is strongly inhibited by carbon monoxide and cytochrome P-450 inhibitors. These results indicate that the enzyme system responsible for non-aromatizing androgen 19-hydroxylase in adrenal is a cytochrome P-450 dependent monooxygenase.

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In order to study the metabolic fate of circulating pregnenolone sulfate (P5S) during pregnancy, 30 mg of newly synthesized tetra (2,2,4,6)-deuterated P5S was administered to the maternal vein 60 minutes prior to cesarean section at term. All pregnant women were volunteers and had been informed of nature of this study. The placenta, maternal urine and blood samples from maternal vein (MV), umbilical cord (U) were collected and deuterated metabolites were analysed by gas chromatography mass spectrometry with a multiple ion detector. Total amounts of metabolites were measured and the ratio of deuterated steroid to the total amounts were calculated (d%). d% of P5S, 16 alpha OH-P5S, 17 alpha OH-P5S, 20 alpha-dihydro-P5S (20P5S), 20P5 and progesterone (P4) in MV were 84.5, 51.6, 95.5, 85.1, 71.2 and 10.9%, respectively. In the placental tissue, 20P5, 20P4 and P4 were also found and d% of these steroids were calculated as 16.1, 3.2 and 3.1%. Only P4 was found with d% of 11.2% in U. In the urine collected for 2 hours after deuterated P5S administration, P5S (40.6%), 20P5S (56.6%) and pregnanediol (34.8%) were identified. Deuterated C19, C18 steroids were not detected in any of the samples studied. When 30 mg of non labeled P5S was also administered to MV at term, the levels of P5S, 20P5S, P4 and 20P4 in MV rose, but the levels of DHA-S were not changed. These results indicate that the circulating P5S in MV can be a precursor of 20P5S, 17P5S, 16P5S, 20P4 and P4, but can not be the precursor of C19, C18 steroid.

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In order to study the mechanism of estriol (E3) formation in the fetoplacental unit, a method for determining E3 and its precursors, 16 alpha-hydroxydehydroepiandrosterone (16 alpha-OH DHA) and 16 alpha-hydroxyestrone (16 alpha-OH E1) in blood using Gas Chromatography-Mass Spectrometry (GC-MS) was newly developed. Deuterium labelled steroids, 3.4.4.-d3-16 alpha-OH DHA 2.4.17.-d3-E2 and 2.4.-d2-E3 were synthesized and used as the internal standards. After removal of free steroids with ether, the residue was solvolyzed and extracted with ethyl acetate as free form. Steroids were analysed after the formation of T.M.S. and T.F.A. derivative. The values for each steroid in maternal peripheral vein blood (M.V.), umbilical artery blood (U.A.) and vein blood (U.V.) were studied. Conjugated steroid values in M.V. obtained at normal vaginal delivery (13 cases) were as follows: 16 alpha-OH DHA 230.3 +/- 76.3 ng/ml, 16 alpha-OH E1 19.34 +/- 7.00 ng/ml and E3 158.8 +/- 50.2 ng/ml (mean +/- S.D.), respectively. The levels of these three steroids in cord blood were significantly higher than those in maternal blood. These results show that the steroids are actively produced in the fetoplacental unit. A significant increase in all the steroids was associated with labor. None of the values for E3 precursors in umbilical blood correlated with that of conjugated E3 in maternal blood. The significance of 16 alpha-OH DHA and 16 alpha-OH E1 as intermediates of E3 formation in the fetoplacental unit is discussed.

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A simple, rapid and highly specific method for the determination of maternal and cord serum 16 alpha-hydroxyestrone (16 alpha-OH E1) during pregnancy and at delivery was developed by mass fragmentographic technique using a Gas chromatography--Mass spectrometry (GC-MS) with application of deuterated estradiol as an internal standard. The equipment used was Shimadzu LKB9000 GC-MS (MID-PM). The T.F.A. derivative of the compounds was analyzed using the GC-MS system equipped with a 1 m glass coil, 1% OV-1 on chromosorb w 60 approximately 80 mesh. The flow rate of carrier gas (helium) was 25 ml/min. The temperatures of column oven, separator and ionization source were kept at 190 degrees C, 270 degrees C and 290 degrees C, respectively. The ionization energy and trap current were 70 eV and 60 microA, respectively. The mass spectrum showed molecular ions at m/ e467 and m/ e478 corresponding to the T.F.A. derivatives of d3-E2 and 16 alpha-OH E1 respectively. Evaluation of the method assessed by recovery experiments was 99.2 percent when 10 ng of 16 alpha-OH E1 was added to 1.0 ml of male serum treated with charcoal. The coefficient of variation was 4.2 percent. Serum conjugated 16 alpha-OH E1 levels during pregnancy increased with progressing gestation. The levels in early pregnancy (5th approximately 15th week), middle pregnancy (16th approximately 27th week) and late pregnancy (28th approximately 42th week) were 1.72 +/- 0.43 ng/ml, 5.35 +/- 2.60 ng/ml, 11.92 +/- 3.93 ng/ml (mean +/- S.D.), respectively. Mean levels of 16 alpha-OH E1 in the umbilical artery were 11.61 +/- 5.17 ng/ml which were statistically higher than these in the maternal peripheral vein (6.06 +/- 4.72 ng/ml) and umbilical vein (9.16 +/- 3.77 ng/ml). Serum 16 alpha-OH E1 levels in the samples obtained from anencephalic pregnancy were significantly lower than those in normal pregnancy, suggesting that fetal adrenal function may be involved in the formation of this steroid during pregnancy.

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A simple and rapid method, latex agglutination inhibition reaction (LAIR), using anti-estriol-16-glucuronide coupled with BSA for estimating estrogen in urine has been developed. Agglutination of the latex reagent by the antiserum occurs within 3 min. The agglutination, however, was completely inhibited by 100 ng/ml or more of estriol. There was a good correlation in results obtained by LAIR and by colorimetry, RIA and GLC. Urine samples (3329) obtained at various gestational weeks were assayed. The samples from 94% of the normal pregnant patients in the third trimester revealed 10 micrograms/ml or more of estrogens. Although the assay is semiquantitative, it may be useful in screening for fetal well-being in high-risk pregnancies.

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A clinical trial of chemoimmunotherapy using cell-wall skeleton of BCG (BCG-CWS) was conducted in 28 patients with acute leukemia in complete remission. Chemotherapy consisted of monthly intensification therapy for 2 months and bimonthly therafter. Immunotherapy with 200 microgram of oil-attached BCG-CWS mixed with 10(7) autochthonous leukemic cells was given intradermally at either of upper or lower extremities every week, except when the patients were on maintenance therapy. No serious systemic side effect attributable to BCG-CWS was noted. Six patients developed mild and transient temparature elevation. Most importnat side effect was local skin reaction. Indulated papules developed in all patients, resulting in draining ulcerations in 26 patients. Increase of immunological reactivity of the patients receiving BCG-CWS was noted. Skin test response to PPD, Varidase, and candida extract showed definite increase. In vitro lymphocyte blastogenic response to PPD, concanavalin-A, and pokeweed mitogen revealed significant increase.

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