Proinflammatory cytokine polymorphisms and the risk of preterm birth and low birthweight in a Japanese population.

Pregnancy and parturition involve a complex and poorly understood molecular and biological interplay between mother and fetus. Inflammatory cytokines have been reported to be associated with fetal growth and parturition. The aim of this study was to examine whether common proinflammatory cytokine polymorphisms are associated with preterm birth (PTB), low birthweight or intrauterine growth restriction in a Japanese population. We assessed a consecutive series of 414 women who had singleton deliveries in Sapporo, Japan between 2001 and 2005. Genotyping of IL1A -889C/T, +4845G/T (A114S), IL1B -511C/T, -31C/T, IL2 -384T/G and IL6 -634C/G polymorphisms was determined by an allelic discrimination assay. The risk of PTB significantly increased in women carrying the IL1A -889T allele (CC genotype [reference]; CT genotype, odds ratios (OR): 2.5; 95% confidence intervals (95% CI): 1.4-4.8; CT+TT genotypes [dominant genotype model], OR: 2.5, 95% CI: 1.3-4.6). Similarly, the risk of PTB significantly increased in women carrying the IL1A +4845T allele (GG genotype [reference]; GT genotype, OR: 2.4, 95% CI: 1.3-4.4; GT+TT genotypes [dominant genotype model], OR: 2.3, 95% CI: 1.2-4.2). The frequency of the IL1A TT haplotype in mothers with PTB was significantly higher than in mothers who had a term birth (P < 0.001), whereas the frequency of the IL1A CG haplotype in mothers who had a PTB was significantly lower (P < 0.001). Our findings suggest that the polymorphisms and haplotypes in the IL1A gene are associated with PTB in Japanese women.

Long-term results of treatments for varicose veins due to greater saphenous vein insufficiency.

AIM: The purpose of this study was to determine the long-term recurrence rates of greater saphenous vein (GSV) insufficiency after treatments for primary varicose veins, and to elucidate risk factors for recurrence. METHODS: This was a multicenter retrospective analysis of 376 limbs of 296 patients treated for primary varicose veins due to GSV insufficiency from January 1996 to December 1997. The recurrence-free rates after stripping surgery, saphenofemoral ligation, and sclerotherapy were estimated. The risk factors for the recurrence of primary varicose veins were estimated by multiple regression analysis. RESULTS: The follow-up period was 3.1+/-1.3 (mean+/-SD) years. The recurrence-free rates at 4 years after stripping, saphenofemoral ligation and sclerotherapy were 80.7%, 64.5%, and 51.3%, respectively. The saphenofemoral ligation group and sclerotherapy group had significantly higher recurrence rates than the stripping group (P=0.002, P<0.001, respectively). There was no difference in recurrence rates between the saphenofemoral ligation group and sclerotherapy group (P=0.074). Logistic regression analysis revealed that being female (P<0.029) and treatment without stripping (P<0.001) increased the recurrence rate. CONCLUSIONS: Stripping surgery may be the treatment of first choice for patients with varicose veins due to GSV insufficiency. Patients who have not received stripping surgery and female patients require closer follow-up.

CYP2D6 ultrarapid metabolizer genotype as a potential modifier of smoking behaviour.

Some 3-10% of Caucasians are deficient in CYP2D6 metabolism (poor metabolizers), due to inheritance of two defective alleles, whereas amplification of the CYP2D6 gene results in ultrarapid metabolism in 1-2% of Caucasian populations. To examine the possible association between CYP2D6 polymorphism and individual smoking behaviour, we analysed the prevalence of CYP2D6 genotypes among 292 long-term heavy smokers, 382 individuals with more variable smoking histories, and 302 never-smokers. The prevalence of ultrarapid metabolizers in heavy smokers (7.9%) was twofold compared to individuals with variable smoking habits (3.7%; odds ratio 2.3, 95% confidence interval 1.2-4.4), and fourfold compared with never-smokers (2.0%) (odds ratio 4.2, 95% confidence interval 1.8-9.8). The frequency of poor metabolizer genotype was approximately 2%, in each smoker group. However, when men and women were studied separately, the prevalence of poor metabolizer genotype was higher in male never-smokers (3.6%) than in variable smokers (2.7%) and heavy smokers (2.2%). Moreover, a trend test, adjusted by age, gender and cancer status, revealed a significant trend for the increased tobacco usage with increased metabolic capacity. Our results are in agreement with the assumption that increased CYP2D6 activity may contribute to the probability of being addicted to smoking.

CYP3A4 allelic variants with amino acid substitutions in exons 7 and 12: evidence for an allelic variant with altered catalytic activity.

OBJECTIVE: To determine the existence of mutant and variant CgammaP3A4 alleles in three racial groups and to assess functions of the variant alleles by complementary deoxyribonucleic acid (cDNA) expression. METHODS: A bacterial artificial chromosome that contains the complete CgammaP3A4 gene was isolated and the exons and surrounding introns were directly sequenced to develop primers to polymerase chain reaction (PCR) amplify and sequence the gene from lymphocyte DNA. DNA samples from Chinese, black, and white subjects were screened. Mutating the affected amino acid in the wild-type cDNA and expressing the variant enzyme with use of the baculovirus system was used to functionally evaluate the variant allele having a missense mutation. RESULTS: To investigate the existence of mutant and variant CgammaP3A4 alleles in humans, all 13 exons and the 5'-flanking region of the human CgammaP3A4 gene in three racial groups were sequenced and four alleles were identified. An A-->G point mutation in the 5'-flanking region of the human CgammaP3A4 gene, designated CgammaP3A4*1B, was found in the three different racial groups. The frequency of this allele in a white population was 4.2%, whereas it was 66.7% in black subjects. The CgammaP3A4*1B allele was not found in Chinese subjects. A second variant allele, designated CgammaP3A4*2, having a Ser222Pro change, was found at a frequency of 2.7% in the white population and was absent in the black subjects and Chinese subjects analyzed. Baculovirus-directed cDNA expression revealed that the CYP3A4*2 P450 had a lower intrinsic clearance for the CYP3A4 substrate nifedipine compared with the wild-type enzyme but was not significantly different from the wild-type enzyme for testosterone 6beta-hydroxylation. Another rare allele, designated CgammaP3A4*3, was found in a single Chinese subject who had a Met445Thr change in the conserved heme-binding region of the P450. CONCLUSIONS: These are the first examples of potential function polymorphisms resulting from missense mutations in the CgammaP3A4 gene. The CgammaP3A4*2 allele was found to encode a P450 with substrate-dependent altered kinetics compared with the wild-type P450.

Behavior of heavy metals in human urine and blood following calcium disodium ethylenediamine tetraacetate injection: observations in metal workers.

To evaluate the effects of calcium disodium ethylenediamine tetraacetate (CaEDTA) on the behavior of 8 heavy metals in human urine and blood, CaEDTA was administered for 1 h by intravenous injection to 18 male metal foundry workers, whose blood lead concentrations (PbB) were between 16 and 59 (mean 34) microg/dl. Significant increases were found in urinary excretion of manganese, chromium, lead, zinc, and copper after the start of CaEDTA injection. Urinary chromium excretion reached a maximal level within 1 h after the start of injection, while urinary manganese, lead, and zinc excretion reached their highest concentrations between 1 and 2 h. Urinary copper excretion reached the highest level between 2 and 4 h. The rapid increases in urinary excretion of five metals were different from the "circadian rhythms," which are the normal, daily variations in renal glomerular filtration, reabsorption, and excretory mechanisms. Plasma lead concentrations were highest 1.5 h after the start of the 1-h injection, while plasma zinc concentration became lowest 5 h after the start of CaEDTA injection. Data suggest that manganese and chromium absorbed in human tissues might be mobilized by CaEDTA.

Prosthetic grafts for above-knee femoropopliteal bypass. A multicenter retrospective study of 564 grafts.

BACKGROUND: Many prosthetic grafts including expanded polytetrafluoroethylene (ePTFE) and polyethylene terephthalate (Dacron) have recently been used for above-knee femoropopliteal bypass. The purpose of this study was to identify the factors affecting patency performance and patient survival. METHODS: A multicenter retrospective analysis of 496 patients who received 564 grafts between 1990 and 1999 (325 ePTFE and 239 Dacron). Follow-up extended to 114.5 months, with a mean of 30.8 months (+/-25.9 months). RESULTS: The overall primary patency rate for all grafts was 71.4% at 5 years, 73.7% for ePTFE, and 68.9% for Dacron grafts. The secondary patency rates at 5 years were 84.1% for ePTFE, and 83.8% for Dacron. No significant differences were found. The logistic regression analysis revealed that younger age at operation and smoking history were correlated with decreased primary patency rate. The patency rates were unaffected by postoperative administration of oral anticoagulants or antiplatelet agents, although pharmacotherapy contributed to the improvement of survival rates. Renal failure, cerebral infarction and Dacron decreased survival rate. CONCLUSIONS: We conclude that the patency performances of prosthetic grafts are satisfying. However, the choice of prosthetic grafts for younger patients or patients with a smoking history need to be carefully considered. Cerebral infarction, chronic renal failure and Dacron grafts may decrease the survival rate. The operative indications should be determined carefully in these cases. The administration of beraprost sodium is recommended for postoperative pharmacotherapy.

Adjustment of creatinine-adjusted value to urine flow rate in lead workers.

Two male lead workers, aged 57 and 51 y, were studied to compare the urinary flow/creatinine-adjusted values published earlier by Araki et al. and by Greenberg and Levine. We collected 24-h urine samples once a month for 31 mo and 16 mo for workers 1 and 2, respectively. The workers' urinary excretions of lead, delta-aminolevulinic acid, and coproporphyrin were measured. No significant correlations between urine flow rate and urinary flow/creatinine-adjusted values published by Araki et al. for the three substances were found for these two workers. However, urinary flow/creatinine-adjusted values presented by Greenberg and Levine for lead and delta-aminolevulinic acid were correlated positively with urine flow rate in the two workers, and their adjusted value for coproporphyrin was correlated positively with urine flow rate in one of the workers. We concluded that use of the urinary flow/creatinine-adjusted value by Greenberg and Levine for biological monitoring poses a problem because of the theoretical fallacy.

[Medical care of AIDS patients by hospitals in Tokyo].

An anonymous questionnaire survey was performed in 1991 in all 749 hospitals in Tokyo, by the Tokyo Metropolitan Government's Acquired Immunodeficiency Syndrome (AIDS) and Human Immunodeficiency Virus (HIV) Infection Research Team, to collect information relating to hospital services for persons with AIDS or HIV infection. The response rate was 61%. The following results were obtained: 1) Sixty-five hospitals (14%) had seen persons with AIDS or HIV infection. Hospitals with a greater number of beds had more experience in seeing persons with AIDS or HIV infection. Thirty-four hospitals (7%) provided outpatient services for patients with AIDS or HIV infection, and 30 hospitals (7%) had the facilities for inpatients. Three hundred forty-seven hospitals (78%) were capable of HIV antibody tests. Ninety-one hospitals (20%) had organized training courses for hospital workers to prevent HIV infection. Two hundred eight hospitals (46%) expressed preference that persons with AIDS or HIV infection be treated at public (non-private) hospitals. 2) The results of multivariate logistic regression analysis indicated that a hospital's acceptance of persons with AIDS or HIV infection for diagnosis or treatment was significantly related to past experience in seeing persons with AIDS or HIV infection, availability of an examination room that protects privacy of patients, presence of a department of internal medicine, and awareness of the availability of special AIDS counselor dispatch services by the Tokyo metropolitan government.

[The epidemiology of prostate cancer--recent trends in prostate cancer incidence and mortality].

Prostate cancer is one of the major malignant diseases in Western countries. In Japan, the incidence and mortality of prostate cancer is not so high, but is continuously increasing. The recent drastic increase in incidence has been attributed to the growth of the elderly population, a westernized diet in daily life, widespread environmental contamination, and improved screening techniques such as the serum PSA test. The epidemiology of prostate cancer hints that its etiology is both environmental and genetic. Androgenic stimulation over time, perhaps due to a high fat diet, has been suggested as a cause of prostate cancer. Dietary factors such as phytoestrogens, vitamins and trace elements are suggested to have a protective effect against prostate cancer, and encourage us to search for means of prevention. Some have suggested that certain polymorphisms increase the risk of prostate cancer, whereas others are searching for genetic mutations that may also increase prostate cancer risk. The cause of prostate cancer is likely to be a combination of environmental and genetic factors.

Maternal folate, alcohol and energy metabolism-related gene polymorphisms and the risk of recurrent pregnancy loss.

Epidemiological studies have suggested that the condition of recurrent pregnancy loss (RPL) may be multifactorial, with both genetic predisposition and environmental factors potentially involved in its pathogenesis. The aim of this study is to elucidate the associations between maternal folate, alcohol and energy metabolism-related gene polymorphisms and the risk of RPL. This case-control study, which involved 116 cases with two or more instances of RPL and 306 fertile controls, was performed in the city of Sapporo, Japan. The associations between eight single nucleotide polymorphisms of folate, alcohol and energy metabolism-related genes [methylenetetrahydrofolate reductase (MTHFR), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), 5-methyltetrahydrofolate-homocysteine methyltransferase reductase (MTRR), alcohol dehydrogenase 1B (ADH1B), aldehyde dehydrogenase 2 (ALDH2), beta-3-adrenergic receptor (ADRB3) and peroxisome proliferator-activated receptor gamma (PPARG)], and RPL were assessed. Without consideration of cigarette smoking or alcohol use, the risk of RPL significantly decreased in women with the MTHFR rs1801133 TT, MTR rs1805087 AG or ALDH2 rs671 AA genotype (P < 0.05). The risk of RPL associated with cigarette smoking and alcohol use decreased significantly in women carrying the MTHFR rs1801133 T allele [odds ratio (OR), 0.51; 95% confidence interval (CI), 0.27-0.95]. Similarly, the risk of RPL significantly decreased in women carrying the MTR rs1805087 G allele (OR, 0.44; 95% CI, 0.23-0.85). Our findings suggest that maternal gene polymorphisms related to folate metabolism may decrease the risk of RPL. Molecular epidemiological studies are needed to unequivocally elucidate the multifactorial effects of both genetic and environmental factors on human fecundity.

Maternal smoking during pregnancy and genetic polymorphisms in the Ah receptor, CYP1A1 and GSTM1 affect infant birth size in Japanese subjects.

Genetic susceptibility to tobacco smoke might have relation to adverse pregnancy outcomes. To estimate the effects of maternal smoking and genetic polymorphisms on infant birth weight and length, we conducted a prospective cohort study of 293 women who delivered singleton live births in Sapporo, Japan. Birth weight and length were significantly lower among infants born to continuously smoking women having the aryl hydrocarbon receptor (AhR) wild type genotype (Arg/Arg; 211 g +/- 76 g; 1.2 cm +/- 0.4 cm, p < 0.01 and p < 0.01, respectively), the CYP1A1 variant genotype (m1/m2 + m2/m2; 170 g +/- 64 g, 0.8 cm +/- 0.3 cm, p < 0.01 and p < 0.05, respectively), or the GSTM1 null genotype (171 g +/- 58 g, 0.6 cm +/- 0.3 cm, p < 0.01 and p < 0.05, respectively). When combinations of these genotypes were considered, birth weight and length were significantly lower for infants of continuously smoking women in the AhR wild type + CYP1A1 variant group (315 g +/- 116 g; 1.7 cm +/- 0.6 cm, p < 0.01 and p < 0.01, respectively) and in the CYP1A1 variant + GSTM1 null group (237 g +/- 92 g; 1.3 cm +/- 0.5 cm, p < 0.05 and p < 0.01, respectively). These genotypes did not confer adverse effects among women who had never smoked; therefore, maternal smoking in combination with maternal AhR, CYP1A1 and GSTM1 genetic polymorphisms may adversely affect infant birth size.

Maternal genetic polymorphisms in CYP1A1, GSTM1 and GSTT1 and the risk of hypospadias.

Hypospadias is one of the most common congenital anomalies. Increased exposure to environmental factors (endocrine-disrupting chemicals and smoking) or maternal endogenous estrogen may cause hypospadias because male sexual differentiation is dependent on normal androgen homeostasis. Moreover, interactions between genetic factors and cigarette smoking and other chemicals have been suggested. It has been demonstrated that the CYP1A1 metabolizes not only environmental chemicals but also estrogens, and glutathione-S-transferases (GSTs) are detoxification enzymes that protect cells from toxicants by conjugation with glutathione. In this study, to investigate the association of CYP1A1 (MspI), GSTM1 and GSTT1 polymorphisms with hypospadias, a case-control study of 31 case mothers who had boys with hypospadias and 64 control mothers was performed in Japan. These polymorphisms were investigated by PCR-based methods using DNA from peripheral lymphocytes. We found that the heterozygous CYP1A1 and heterozygous and homozygous CYP1A1 were less frequent in the case mothers than in the control mothers [adjusted odds ratio (OR)=0.17, 95% confidence interval (CI)=0.04-0.74, OR = 0.28, 95% CI = 0.08-0.97, respectively]. We found no effect of maternal smoking on the hypospadias risks among the gene polymorphisms. The results suggest that mothers with the CYP1A1 MspI variant allele may have a decreased risk for hypospadias.

Adjustment for urinary flow rate: an improved approach to biological monitoring.

The use of urinary monitoring in medical surveillance programs in industry requires the development of an accurate and unbiased index of urinary concentrations of occupational toxins. To examine the effects of urinary flow rate on adjusted and non-adjusted urinary excretion of 11 heavy metals and organic substances, 19 metal-foundry workers were studied during four time periods of the day under conditions of water restriction and loading. The results indicate that urinary flow rate significantly affects not only the non-adjusted urinary concentration for all substances, but also affects timed excretion as well as concentrations adjusted to urinary specific gravity and to urinary creatinine during any time period of the day. On the other hand, the concentration adjusted to urinary flow rate (UF-adjusted concentration) is independent by definition of urinary flow; therefore, this adjustment is applicable for highly diluted and highly concentrated urine samples without repetition of urine collection. It is concluded that UF-adjusted concentration is a useful index for the measurement of most urinary substances, while adjustments to urinary specific gravity and to urinary creatinine concentration have only limited utility in evaluating toxin levels in spot urine samples.

Adjustment of creatinine-adjusted values in urine to urinary flow rate: a study of eleven heavy metals and organic substances.

The use of urinary monitoring in medical surveillance programs in industry requires development of an accurate and unbiased index of urinary concentrations of occupational toxins. To examine the effects of urinary flow (UF) rate on the standard creatinine (Cn)-adjusted value and the UF- and Cn-adjusted values according to Greenberg and Levine and to Araki et al. for 11 heavy metals and organic substances, 19 metal-foundry workers aged 34-59 years (mean 48), who had been exposed to lead, zinc, and copper for 2-17 (mean 10) years, were studied during four periods of the day under water-free, water-restrictive and water-loading conditions for 6 days. The blood lead concentrations of the 19 workers ranged from 22 to 59 (mean 38) micrograms/dl. The results indicated that there was no significant UF effect on the UF- and Cn-adjusted values of Araki et al. for all urinary substances except mercury and coproporphyrin. On the other hand, the UF- and Cn-adjusted value of Greenberg and Levine was positively correlated with UF rate for all urinary substances, and the standard Cn-adjusted value was either positively or inversely correlated with UF rate for many urinary substances. Therefore, the UF- and Cn-adjusted value of Araki et al. is considered to be applicable to the measurement of most urinary substances under conditions of wide variation in UF rate.

Changes in natural killer cell subpopulations in lead workers.

To investigate the effects of lead on human immune system, we analyzed T cell subpopulations (CD4+, CD8+ and CD3+ cells), natural killer (NK) cell subpopulations (CD16+ and CD57+ cells) and B (CD19+) cells in peripheral blood in 29 male lead workers. All were engaged in manufacturing lead stearate in a chemical factory. They were aged 23-74 (mean 49) years. Their blood lead concentrations (PbB) were between 7 and 35 (mean 18) micrograms/dl. They were divided into two groups according to their PbB: a high-PbB group (> or = 20 micrograms/dl), and a low-PbB group (< 20 micrograms/dl). The control group consisted of 19 "healthy" male workers without a history of occupational exposure to lead or to other hazardous substances, aged 48-67 (mean 58) years. The number and percentage of CD16+ cells in the high-PbB group were significantly lower than those in the controls and in the low-PbB group. There was significant negative correlation between the number of CD16+ cells and PbB in the lead workers. The percentage of CD8+ cells in the high-PbB group was larger than that in the controls and in the low-PbB group. It is suggested that the CD16+ NK cell should be a major site of the effects of lead on lymphocyte subpopulations.

Changes in T cell subpopulations in lead workers.

To investigate the effects of lead on the human immune system, we analyzed T cell subpopulations and B (CD19+) cells in peripheral blood in 71 male lead workers. They were engaged in manufacturing lead stearate in a chemical factory, aged 20 to 74 (mean 48) years. Their blood lead concentrations (PbB) were between 7 and 50 (mean 19) micrograms/dl. The control group consisted of 28 "healthy" male volunteers without a history of occupational exposure to lead or other hazardous substances, aged 33 to 67 (mean 55) years. In comparison with the controls, a significant reduction in the number of CD3+CD45RO+ (memory T) cells and a significant expansion in the percentage of CD8+ cells in the lead workers were found. There was a significant positive correlation between the percentage of CD3+CD45RA+ (naive T) cells and PbB in the lead workers. It is suggested that CD45RO+ memory T cells may be most susceptible to the effects of lead on T cell subpopulations.

Analysis of toxic gas produced by heating tar epoxy resin paint to assess work atmosphere.

Two male workers were acutely intoxicated with gas produced by heating tar epoxy resin paint, and developed peripheral neuropathy. To assess the work atmosphere, we analyzed the degradation products by GC-MS. The major toxic products emitted by heating tar epoxy resin were hydrogen cyanide, phenol, and benzene, as well as naphthalene. From 1 m2 of the surface of steel plates painted with tar epoxy resin, 2.4 g of hydrogen cyanide, 9.6 g of benzene, and 1.2 g of nephthalene were produced by heating at 1000 degrees C, At 700 degrees C, the amounts of phenol and p-isopropylphenol produced were 3.7 g and 0.57 g, respectively. Based on these results and the area of steel surface burned, the concentration of hydrogen cyanide, benzene, and phenol in the atmosphere of work environment was estimated to be 16, 64 and 24 mg/m3, respectively. Some of the symptoms of the workers including peripheral neuropathy might be related to the sole or cooperative action of the foregoing toxic chemicals mentioned above.