The relationship between new stroke onset and serum thiocyanate as an indicator to cigarette smoking.

The case subjects were 67 stroke patients (27 males and 40 females, mean age 65.7+/-7.1), who suffered from stroke attack and had participated in annual medical check-up between 1987 and 1988 at Oyabe Public Health Center. The controls, which were adjusted for sex, age and blood pressure level with the case subjects, were selected from participants attending their annual medical check-up in 1987-1988. The serum thiocyanate level of case subjects were significantly higher than that of controls (p<0.05). The usefulness of measurement of serum thiocyanate level as an indicator of smoking status was assessed by using multiple logistic regression analysis adjusting for body mass index, mean blood pressure, and serum cholesterol was found. An odds ratio of 3.00 (95% confidence interval: 1.06-8.48, p<0.05) in cerebral infarction. It is considered that serum thiocyanate may be useful as an indicator of smoking status to assess the relationship with stroke onset, especially for cerebral infarction.

Presence of nitrate-accumulating sulfur bacteria and their influence on nitrogen cycling in a shallow coastal marine sediment.

Nitrate flux between sediment and water, nitrate concentration profile at the sediment-water interface, and in situ sediment denitrification activity were measured seasonally at the innermost part of Tokyo Bay, Japan. For the determination of sediment nitrate concentration, undisturbed sediment cores were sectioned into 5-mm depth intervals and each segment was stored frozen at -30 degrees C. The nitrate concentration was determined for the supernatants after centrifuging the frozen and thawed sediments. Nitrate in the uppermost sediment showed a remarkable seasonal change, and its seasonal maximum of up to 400 microM was found in October. The directions of the diffusive nitrate fluxes predicted from the interfacial concentration gradients were out of the sediment throughout the year. In contrast, the directions of the total nitrate fluxes measured by the whole-core incubation were into the sediment at all seasons. This contradiction between directions indicates that a large part of the nitrate pool extracted from the frozen surface sediments is not a pore water constituent, and preliminary examinations demonstrated that the nitrate was contained in the intracellular vacuoles of filamentous sulfur bacteria dwelling on or in the surface sediment. Based on the comparison between in situ sediment denitrification activity and total nitrate flux, it is suggested that intracellular nitrate cannot be directly utilized by sediment denitrification, and the probable fate of the intracellular nitrate is hypothesized to be dissimilatory reduction to ammonium. The presence of nitrate-accumulating sulfur bacteria therefore may lower nature's self-purification capacity (denitrification) and exacerbate eutrophication in shallow coastal marine environments.

Serum thiocyanate concentration as an indicator of smoking in relation to deaths from cancer.

All residents aged 40 years or more in Oyabe City, Toyama Prefecture, Japan were involved in an annual medical check-up between 1987 and 1988. The cohort was followed and death certificates from cancers were confirmed prospectively. During follow-up to December 31 st, 1994, 100 deaths (28 gastric, 17 lung and 55 other cancers) from cancers occurred, and these subjects were included in this study as the case group. Subjects in the control group, matched for gender and age with the cases, were selected randomly from participants whose serum samples had been stocked during annual medical check-up. The concentration of serum thiocyanate in all (79.8 µmol/l), gastric (86.7 µmol/l) and lung (90.0 µmol/l) cancer patients were significantly higher than that of relevant controls (64.3 µmol/l, 59.0 µmol/l and 61.0 µmol/l, respectively; and p<0.001, p<0.001 and p<0.05, respectively). After adjusting for BMI, blood pressure and total serum cholesterol, the results of multiple logistic regression analysis showed that the risk of all cancers (OR=3.40, 95% confidence interval (95% Cl): 1.67-6.96, p<0.01), gastric cancer (OR=7.98, 95% CI: 1.91-33.34, p<0.05) and lung cancer (OR=8.83, 95% CI: 1.19-65.65, p<0.05) were elevated significantly with logarithm transformed values of serum thiocyanate increased. The present findings suggested that in epidemiological studies confirmation of smoking status with biomarkers such as serum thiocyanate may be important, although considering the small sample size, a relatively weaker risk to interested factors rather than the strong relationship between smoking and cancer was noted.

Purification and DNA-binding properties of the cro-type regulatory repressor protein cng encoded by the Lactobacillus plantarum phage phi g1e.

The putative repressor protein Cng (10kDa on an SDS gel) for the lytic pathway of Lactobacillus plantarum phage φg1e was purified using the Escherichia coli Pt7 system, and its DNA-binding ability for the seven operator-like sequences, the GATAC-boxes (Gb1 to Gb7), was investigated in vitro. In gel-shift assays, Cng selectively bound to the DNA fragments containing the GATAC-box(es). In addition, DNase I footprinting analysis with supercoiled DNA demonstrated that Cng can specifically cover about a 25bp region centered around each of the GATAC-boxes, although two boxes, Gb4 and Gb6, were only partially protected. Moreover, protein crosslinking experiments using glutaraldehyde suggested that Cng most likely functions as a dimer. On the other hand, the binding ability of Cpg for the GATAC-boxes in supercoiled DNA was also examined under the same conditions as in Cng; unlike Cng, Cpg covered Gb4 and Gb6 completely sufficiently as well as the other five boxes. Thus, the present and previous [Kakikawa et al., Gene 215 (1998) 371-379; 242 (2000) 155-166] results indicate a possibility that the two proteins Cng and Cpg selectively bind to the GATAC-boxes that act as operators, and can decide between the lytic or lysogenic pathways through repression of the promoter activity of P(R) as well as P(L).

Purification and biochemical properties of an N-hydroxyarylamine O-acetyltransferase from Escherichia coli.

The N-hydroxyarylamine O-acetyltransferase of Escherichia coli has been expressed as a histidine tagged fusion protein and purified using immobilized nickel column chromatography. The molecular mass of the histidine tagged N-hydroxyarylamine O-acetyltransferase was estimated to be 60.0 kDa by gel filtration and 34.0 kDa by SDS-PAGE and DNA sequence, suggesting that the native enzyme exists as homo dimer. The catalytic properties were investigated using o-aminobenzoic acid as a substrate. No difference in acetyltransfer activity was observed between histidine tagged protein and untagged enzyme. Kinetic studies indicated a ping-pong bi bi mechanism of the catalysis. Inhibition by N-ethylmaleimide and salicylic acid was competitive with o-aminobenzoic acid and non-competitive with acetyl-CoA.

Clinical efficacy of peritoneovenous shunting for the treatment of severe ovarian hyperstimulation syndrome.

We investigated prospectively the clinical efficacy of a newly developed continuous autotransfusion system of ascites (CATSA) without protein supplement in patients with severe ovarian hyperstimulation syndrome (OHSS). Peritoneovenous shunting was used to recirculate ascites. The CATSA was performed for 5 h at a rate of 100-200 ml/h once a day. Eighteen patients were treated with the CATSA (CATSA group) and 36 were treated with an intravenous 37.5 g/day of albumin supplement (albumin group). Hospital stay was significantly shorter in the CATSA group than in the albumin group (10.0 +/- 5.7 versus 13.9 +/- 6.2 days, P < 0.01). Haematocrit value reached <40% significantly earlier in the CATSA group (on hospital days 3.9 +/- 3.2 versus 5.9 +/- 2.5, P < 0.01). Using a single procedure, haemoconcentration, urinary output and pulse pressure were markedly improved in the CATSA group compared with the albumin group. Discomfort due to massive ascites diminished promptly and did not recur in nine of 18 CATSA group patients, whereas it persisted in all 36 patients in the albumin group. The serum concentration of protein was maintained in the CATSA group, whereas it did not increase in the albumin group despite daily supplementation with 37. 5 g of albumin. Apparent adverse effects of each procedure were not observed in either group. The mean values of several parameters in the serum pertinent to the coagulation-fibrinolysis system did not change significantly in either group after the procedure. It was concluded that the CATSA procedure expanded circulating plasma volume without exogenous albumin and appeared to lead to a prompt recovery from severe conditions of OHSS.

Association of a decrease in antithrombin III activity with a perinatal elevation in aspartate aminotransferase in women with twin pregnancies: relevance to the HELLP syndrome.

BACKGROUND/AIMS: Decreased antithrombin III (AT-III) activity and/or thrombocytopenia associated with an elevated serum level of aspartate aminotransferase in late pregnancy can threaten the lives of both the mother and the fetus. We investigated whether antenatal declines in AT-III activity and the platelet count occur in late twin pregnancy and whether reduced AT-III activity and/or thrombocytopenia precedes an increase in the serum level of aspartate aminotransferase. METHODS: The platelet count, AT-III activity, and the serum level of aspartate aminotransferase were determined weekly or biweekly in 237 women with twin pregnancies in a longitudinal and partly prospective study. RESULTS: Both AT-III activity and the platelet count decreased gradually in the last month of pregnancy, irrespective of the presence or absence of clinical signs of pre-eclampsia. A perinatal elevation in aspartate aminotransferase occurred in 36 (15%) of 237 women. The risk of a perinatal elevation in aspartate amino-transferase increased as the antenatal AT-III activity and/or the platelet count decreased. Pre-eclampsia developed in 60 women (25%). The relative risk of a perinatal aspartate aminotransferase elevation (95% confidence interval) for the 60 women with pre-eclampsia, the 60 women with a platelet count < or = the 25th percentile (164 x 10(9)/1), and the 60 women with AT-III activity < or = the 25th percentile (76% of normal) was 1.9 (1.0 to 3.4), 4.1 (2.3 to 7.5), and 5.9 (3.2 to 11.1), respectively, compared with the remaining 177 women. CONCLUSIONS: AT-III activity and platelet count gradually decreased in the last month of twin pregnancies. A perinatal aspartate aminotransferase elevation was preceded by marked decreases in these parameters in women with twin pregnancies. The monitoring of AT-III activity and platelet count in women who exhibit a gradual decline in these parameters may help to avoid the development of severe HELLP syndrome.

Mutagenicities of 2,4- and 2,6-dinitrotoluenes and their reduced products in Salmonella typhimurium nitroreductase- and O-acetyltransferase-overproducing Ames test strains.

Mutagenicities of 2,4- and 2,6-dinitrotoluene (2,4-and 2,6-DNT), and reduced metabolites formed by the incubation of 2,4- and 2,6-DNT with Salmonella typhimurium TA98, were tested using S. typhimurium YG strains possessing high level of nitroreductase (NR) and/or O-acetyltransferase (OAT) activities. All compounds tested showed greatest mutagenic activities toward strains YG1041 and YG1042, which possess high levels of NR and OAT activities. The relative mutagenic activities of 2,4-DNT and its related compounds toward YG1041 and YG1042 were aminonitrotoluenes<2,4-DNT<2,2'-dimethyl-5, 5'-dinitroazoxybenzene (2,2'-DM-5, 5'-DNAOB)4-hydroxylamino-2-nitrotoluene (4HA2NT)<<4, 4'-dimethyl-3,3'-dinitroazoxybenzene (4,4'-DM-3,3'-DNAOB), and aminonitrotoluenes (2A4AT, 4A2NT)<2,4-DNT<4HA2NT4,4'-dimethyl-3, 3'-dinitroazoxybenzene (4,4'-DM-3,3'-DNAOB)<2HA4NT, respectively. In addition, the relative mutagenic activities of 2,6-DNT and its related compounds toward YG1041 and YG1042 were 2, 6-DNT<2-hydroxylamino-6-nitrotoluene (2HA6NT)<2,2'-dimethyl-3, 3'-dinitroazoxybenzene (2,2'-DM-3,3'-DNAOB), and 2-amino-6-nitrotoluene (2A6NT)<2,6-DNT<2HA6NT, respectively. These results, together with previous findings, suggested that aminohydroxylamino dimethylazoxybenzenes or aminohydroxylamino dimethylazobenzenes produced either by the reduction of hydroxylaminonitrotoluenes or by the reduction of dimethyl dinitroazoxybenzenes are active metabolites responsible for the mutagenic activities of 2,4- and 2,6-DNT.

Lower risks of adverse outcome in twins conceived by artificial reproductive techniques compared with spontaneously conceived twins.

The outcomes of twins conceived by 136 women after medical assistance (MA) such as ovulation induction with or without assisted reproductive techniques, and twins conceived spontaneously (SP) by 72 women were compared. All 208 women were monitored from < 20 weeks gestation; they all delivered at > or = 24 weeks gestation. The chorionicity of the placenta was diagnosed antenatally and confirmed after delivery. There were 10 perinatal deaths; the physical and neurological status of the remaining 406 infants was assessed at 1 year of corrected age. There were no differences in gestational age at birth, the birth weights of the larger and smaller twins, the birth weight discordance, or the incidence of life-threatening major malformations between groups. Adverse infant outcomes, such as death, cerebral palsy and mental retardation occurred in nine (3.3%) of 272 MA twins compared with 12 (8.3%) of 144 SP twins (P < 0.05). The placenta was monochorionic in only three (2.2%) of 136 MA twin pregnancies compared with 41 (57%) of 72 SP twin pregnancies (P < 0.001). Of the 21 infants with adverse outcomes, nine had monochorionic placentas. Thus, the risk of an adverse outcome was 2.8-fold higher (95% confidence interval (CI) 1.2-6.4) in monochorionic twins than in dichorionic twins (10 versus 3.7%; P < 0.05). There was no difference in the incidence of adverse infant outcomes between SP (4.8%) and MA (3.4%) twins with dichorionic placentas. These findings suggest that ovulation induction in itself was not associated with an adverse outcome of twin pregnancies. The lower frequency of monochorionic placentas in MA twins may have been responsible for the lower risk of an adverse outcome in MA twins.

Synthesis of pyridinium dinitrobenzyl sulfates and potassium (dinitrobenzyl beta-D-glucopyranosid)uronates.

Sulfates and glucuronides of 2,4-dinitrobenzyl alcohol 1a and 2,6-dinitrobenzyl alcohol 1b, which are major or putative metabolites of 2,4-dinitrotoluene (2,4-DNT) and 2,6-dinitrotoluene (2,6-DNT), were synthesized from 1a and 1b by reaction with pyridinium sulfonate and methyl (2,3,4-tri-O-acetyl-alpha-D-glucopyranosyl)uronate bromide 3, respectively, as their pyridinium salts (2a, 2b) and potassium salts (6a, 6b). These conjugates are important for the study of the carcinogenicity of 2,4-DNT and 2,6-DNT.

Biliary excretion and microfloral transformation of major conjugated metabolites of 2,4-dinitrotoluene and 2,6-dinitrotoluene in the male Wistar rat.

1. Major biliary conjugates of the male Wistar rat dosed orally with 2,4-dinitrotoluene (2,4-DNT) or 2,6-dinitrotoluene (2,6-DNT) were examined by hplc using potassium 2,4-dinitrobenzyl glucuronide (potassium 2,4-DNB-G), potassium 2,6-dinitrobenzyl glucuronide (potassium 2,6-DNB-G), pyridinium 2,4-dinitrobenzyl sulphate (pyridinium 2,4-DNB-S) and pyridinium 2,6-dinitrobenzyl sulphate (pyridinium 2,6-DNB-S) as authentic compounds. Other metabolites were also examined by hplc. In addition, metabolites formed by incubation of potassium 2,4-DNB-G and potassium 2,6-DNB-G with rat intestinal microflora under nitrogen were examined by hplc. 2. Conjugates detected directly from bile following administration of 2,4-DNT and 2,6-DNT were 2,4-DNB-G and 2,6-DNB-G, which accounted for 35.0 and 51.5% of the administered dose respectively. No peaks corresponding to pyridinium 2,4-DNB-S and pyridinium 2,6-DNB-S were detected in bile samples. 3. 2-Amino-4-nitrotoluene, 4-amino-2-nitrotoluene, 2,4-diaminotoluene and 4-acetylamino-2-nitrobenzoic acid (0.02-0.12% of the dose excreted in 24 h), in addition to the known metabolites 2,4-dinitrobenzyl alcohol (2,4-DNB), 2,4-dinitrobenzaldehyde and 2,4-dinitrobenzoic acid (0.09-0.14%), were detected in ether extracts of bile of rat given 2,4-DNT. 2,6-Dinitrobenzyl alcohol (2,6-DNB), 2-amino-6-nitrotoluene and 2,6-dinitrobenzaldehyde (0.02-0.03%), which are known metabolites, were detected in ether extracts of bile from rat given 2,6-DNT. 4. Potassium 2,4-DNB-G was transformed by the anaerobic incubation of rat intestinal microflora into 2,4-DNB, 4-amino-2-nitrobenzyl alcohol and 2-amino-4-nitrobenzyl alcohol. Potassium 2,6-DNB-G was transformed into 2,6-DNB and 2-amino-6-nitrobenzyl alcohol by the anaerobic incubation. Time-course studies showed that 2,4-DNB, 4-amino-2-nitrobenzyl alcohol, 2-amino-4-nitrobenzyl alcohol and 2,6-DNB, 2-amino-6-nitrobenzyl alcohol peaked at 30, 75, 120 and 10, 50 min respectively. 5. These results, together with previous findings, show that 2,4-dinitrobenzaldehyde and 2,6-dinitrobenzaldehyde, which are potent mutagens, are formed either by the hepatic metabolism of 2,4-DNB and 2,6-DNB formed by the intestinal metabolism of 2,4-DNB-G and 2,6-DNB-G excreted in bile or by the direct hepatic metabolism of 2,4-DNT and 2,6-DNT.

The clinical efficacy of gamete intrafallopian transfer by minilaparotomy versus in vitro fertilization and embryo transfer.

OBJECTIVE: To evaluate the clinical performance of gamete intrafallopian transfer (GIFT) and in vitro fertilization and embryo transfer (IVF-ET). METHODS: Infertile women were divided into 2 groups: for GIFT, 239 patients (326 cycles) with at least 1 patent tube; and, for IVF-ET, 125 patients (210 cycles) with bilateral tubal occlusion. A specially designed retractor was developed to replace the gametes into an appropriate section of the Fallopian tube accurately and safely. Several parameters, including the pregnancy and delivery rates of each group, were compared. RESULTS: The success rate per trial in the GIFT group was approximately 1.5 times higher than that in the IVF group (pregnancy rate: 44.2% vs. 31.0%, p < 0.01; delivery rate: 33.4% vs. 22.9%, p < 0.01). The pregnancy and delivery rates of GIFT decreased steadily with the number of trials. These apparent decreases were not observed up to the 3rd trial in IVF-ET cases. CONCLUSIONS: GIFT with a minilaparotomy procedure yielded significantly higher success rates than IVF-ET. Accordingly, GIFT is considered to be the first treatment choice in infertility cases with at least 1 patent Fallopian tube.

The problem of relating fetal outcome with breech presentation to mode of delivery.

We retrospectively analyzed 546 consecutive singleton pregnancies with breech presentations that ended at > or = 36 weeks of gestation for the relationship between the intended mode of delivery and fetal outcome. Twelve patients were excluded from the analysis because these infants had major malformations. Of the 534 remaining patients, 124 (23%) were delivered by elective cesarean section. The other 410 women (77%) went into spontaneous labor. Intrapartum emergency cesarean section was required in 112 (27%) of these 410 women; the other 298 (73%) were delivered vaginally. There were 5 poor neonatal outcomes: 3 perinatal deaths and 2 cases of cerebral palsy probably due to intrapartum asphyxia. The risk of poor outcome was thus 1.2% (5/410), in the intended vaginal delivery group vs. no such outcome in the group of 124 patients that had an elective cesarean section. Three of 5 infants with poor outcome were actually born by emergency cesarean section and comparisons of results according to ultimate method of delivery rather than according to intended method of delivery may be misleading and in our case would have been biased against cesarean section.

The effect of managed care on the treatment outcome of substance use disorders.

This study examined the effect of managed care and other reimbursement mechanisms on the outcome of substance abuse treatment at a single treatment facility. A retrospective review of 1594 patient records yielded treatment utilization, diagnostic, and demographic data. Recidivism rates for intensive managed care, traditional managed care, private pay, and state-funded groups of patients were compared. Results showed that, contrary to expectations, recidivism rates were not different for managed vs nonmanaged care patients. In addition, recidivist patients had significantly more ICD-9 diagnoses than nonrecidivist patients. A discussion of future research suggests that other outcome measures need to be examined in addition to recidivism rate, such as psychosocial functioning following treatment and indicator(s) of severity of illness, to better determine the effect of managed care and other reimbursement mechanisms on treatment outcome.

Enzyme-cytochemistry of human chorion laeve at term: enzyme localization on the chorionic trophoblast.

Is the chorion laeve merely a remnant of the chorion frondosum in placental development? Or is it metabolically active, having something to do with maternofetal interactions? In order to answer these questions at least in part, we determined the ultracytochemical localizations of some important enzymes such as nonspecific phosphatase (alkaline phosphatase), specific phosphatase (Ca(++)-ATPase and 5'-nucleotidase) and adenylate cyclase in the human chorion laeve at term. Strong activities of these enzymes were localized by ultracytochemistry on the plasma membrane of the trophoblast in the chorion laeve. These enzyme activities were confirmed by a series of cytochemical-control experiments, i.e., substrate-free control, heat-stability control, and inhibition control by inhibitors of alkaline phosphatase. These observations indicate that the chorionic trophoblast is probably metabolically active and that it might play an important role in the physiology of the fetal membrane.

Enzyme-cytochemistry of human chorion laeve at term: enzyme localization on the chorionic trophoblast.

Is the chorion laeve merely a remnant of the chorion frondosum in placental development? Or is it metabolically active, having something to do with maternofetal interactions? In order to answer these questions at least in part, we determined the ultracytochemical localizations of some important enzymes such as nonspecific phosphatase (alkaline phosphatase), specific phosphatase (Ca(++)-ATPase and 5'-nucleotidase) and adenylate cyclase in the human chorion laeve at term. Strong activities of these enzymes were localized by ultracytochemistry on the plasma membrane of the trophoblast in the chorion laeve. These enzyme activities were confirmed by a series of cytochemical-control experiments, i.e., substrate-free control, heat-stability control, and inhibition control by inhibitors of alkaline phosphatase. These observations indicate that the chorionic trophoblast is probably metabolically active and that it might play an important role in the physiology of the fetal membrane.