Environmental radiation at Izu-Oshima after the Fukushima Daiichi nuclear power plant accident.

Environmental radiation at Izu-Oshima Island was observed 6 months after the accident at the Fukushima Daiichi Nuclear Power Plant (F1-NPP). A car-borne survey of the dose rate in air was conducted over the entire island and the results were compared with measurements performed in 2005 (i.e. before the accident). The activity concentrations of (134)Cs and (137)Cs were also measured using a germanium detector. The dose rate in air was found to be 2.9 ± 1.2 times higher than that in 2005 and (134)Cs was detected on Izu-Oshima Island. These results are attributed to the accident at the F1-NPP.

Association between methylene tetrahydrofolate reductase and glutathione S-transferase M1 gene polymorphisms and chronic myeloid leukemia in a Brazilian population.

Chronic myeloid leukemia is a hematopoietic stem cell disorder that causes uncontrolled proliferation of white blood cells. Although the clinical and biological aspects are well documented, little is known about individual susceptibility to this disease. We conducted a case-control study analyzing the prevalence of the polymorphisms MTHFR C677T, MTHFR A1298C, del{GSTM1}, del{GSTT1}, and haptoglobin in 105 patients with chronic myeloid leukemia (CML) and 273 healthy controls, using PCR-based methods. A significant association with risk of developing CML was found for MTHFR 1298AA (odds ratio (OR) = 1.794; 95% confidence interval (CI) = 1.14-2.83) and GSTM1 non-null (OR = 1.649; 95%CI = 1.05-2.6) genotypes, while MTHFR 1298AC (OR = 0.630; 95%CI = 0.40-0.99) and GSTM1 null (OR = 0.606; 95%CI = 0.21-0.77) genotypes significantly decreased this risk. There appeared to be selection for heterozygosity at the MTHFR 1298 locus. The considerable range of variation in this and other human populations may be a consequence of distinctive processes of natural selection and adaptation to variable environmental conditions. The Brazilian population is very mixed and heterogeneous; we found these two loci to be associated with CML in this population.

Evidence for an association between haptoglobin and MnSOD (Val9Ala) gene polymorphisms in essential hypertension based on a Brazilian case-control study.

Essential hypertension is a complex and multifactorial trait; genetic and environmental factors interact to produce the final phenotype. Studies have demonstrated association of hypertension with varied gene polymorphisms. However, demonstration of common genetic causes in the general population remains elusive. We investigated a possible association between hypertension and haptoglobin, angiotensin I-converting enzyme (ACE), glutathione S-transferases GSTM1 and GSTT1, MnSOD (Val9Ala), CAT (-21A/T), and GPX1 (Pro198Leu) gene polymorphisms in an urban Brazilian population group from Brasília. Although ACE has been reported to be one of the main polymorphisms associated with hypertension, we found no association with ACE's specific genotypes. However, a possible association with Hp1-1 and MnSOD Val/Ala genotypes suggests that, at least for the Brazilian population, polymorphisms related to oxidative stress should be more deeply investigated.

Antioxidant effect of haptoglobin phenotypes against DNA damage induced by hydrogen peroxide in human leukocytes.

Human haptoglobin is classified into three major phenotypes: Hp1-1, Hp2-1 and Hp2-2; there are two autosomal alleles Hp*1 and Hp*2, and the Hp*1 allele has two subtypes, Hp*1F and Hp*1S. Haptoglobin acts as an antioxidant, preventing hemoglobin-driven oxidative damage. We used the comet assay to examine oxidative damage to DNA induced by hydrogen peroxide in human leukocytes; we also looked for differences in the antioxidant capacity of haptoglobin subtypes. Haptoglobin genotypes were determined through allele-specific polymerase chain reaction, visualized on a polyacrylamide gel. The Hp1-1 genotype had the least DNA damage; this indicates that Hp alleles differ in their protective effects against oxidative damage. Among Hp*1 alleles, Hp*1F was the most protective.

Dietary carotenoid-rich pequi oil reduces plasma lipid peroxidation and DNA damage in runners and evidence for an association with MnSOD genetic variant -Val9Ala.

Physical training induces beneficial adaptations; however, exhausting exercise increases reactive oxygen species generation, resulting in damage to DNA and tissues. Pequi (Caryocar brasiliense), a fruit of the Brazilian Cerrado, contains a carotenoid-rich oil. We investigated whether pequi oil had antioxidant effects in runners. Evaluations were made after outdoor races before and after ingestion of 400 mg pequi-oil capsules for 14 days. Blood samples were taken after races and submitted to comet and TBARS assays and biochemical analyses of creatine kinase (CK), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). To determine if the protective effects of pequi-oil were influenced by antioxidant enzyme genotypes, MnSOD (-Val9Ala), CAT (-21A/T) and GPX1 (Pro198Leu) gene polymorphisms were also investigated. Pequi oil was efficient in reducing tissue injuries evaluated for AST and ALT, particularly in women, and in reducing DNA damages in both sexes. Except for CK levels, the results were influenced by MnSOD genotypes; heterozygous excess was related to less DNA damage, tissue injury and lipid peroxidation, besides presenting a better response to pequi oil against exercise-induced damage.

Genetic variability in maned wolf based on heterologous short-tandem repeat markers from domestic dog.

The maned wolf (Chrysocyon brachyurus) is the largest South American canid. Habitat loss and fragmentation, due to agricultural expansion and predatory hunting, are the main threats to this species. It is included in the official list of threatened wildlife species in Brazil, and is also protected by IUCN and CITES. Highly variable genetic markers such as microsatellites have the potential to resolve genetic relationships at all levels of the population structure (among individuals, demes or metapopulations) and also to identify the evolutionary unit for strategies for the conservation of the species. Tests were carried out to verify whether a class of highly polymorphic tetranucleotide repeats described for the domestic dog effectively amplifies DNA in the maned wolf. All five loci studied were amplified; however, one of these, was shown to be monomorphic in 69 maned wolf samples. The average allele number and estimated heterozygosity per polymorphic locus were 4.3 and 67%, respectively. The genetic variability found for this species, which is considered threatened with extinction, showed similar results when compared to studies of other canids.

Genetic variability in maned wolf based on heterologous short-tandem repeat markers from domestic dog

The maned wolf (Chrysocyon brachyurus) is the largest South American canid. Habitat loss and fragmentation, due to agricultural expansion and predatory hunting, are the main threats to this species. It is included in the official list of threatened wildlife species in Brazil, and is also protected by IUCN and CITES. Highly variable genetic markers such as microsatellites have the potential to resolve genetic relationships at all levels of the population structure (among individuals, demes or metapopulations) and also to identify the evolutionary unit for strategies for the conservation of the species. Tests were carried out to verify whether a class of highly polymorphic tetranucleotide repeats described for the domestic dog effectively amplifies DNA in the maned wolf. All five loci studied were amplified; however, one of these, was shown to be monomorphic in 69 maned wolf samples. The average allele number and estimated heterozygosity per polymorphic locus were 4.3 and 67%, respectively. The genetic variability found for this species, which is considered threatened with extinction, showed similar results when compared to studies of other canids.

A new delivery system for 5-fluorouracil using prodrug and converting enzyme.

AIMS: To evaluate a new delivery system of 5-fluorouracil (5-FU) using 5-fluorocytosine (5-FC) as a prodrug and cytosine deaminase induced in vitro and in vivo. METHODS: Fibroblastic cells from rabbit Tenon's capsule were cultured. The cells were exposed to 5-FU and 5-FC with or without cytosine deaminase induced by recombinant adenovirus. In the in vitro study, cell proliferation and DNA synthesis were assessed by MTS, BrdU assay. The effect of 5-FC removal after the treatment of 5-FC and cytosine deaminase induction was also assayed. In the in vivo study cells with or without cytosine deaminase induction were transplanted into the subconjunctival space of mice, followed by eye drops of 1000 microg/ml of 5-FC three times a day. The mice were sacrificed at days 1, 5, and 10, then the cells transplanted were evaluated. RESULTS: Cell proliferation was inhibited by exposure to 5-FU in a dose dependent manner; however, up to 1000 microg/ml of 5-FC did not affect cell proliferation. Cell proliferation was inhibited by exposure to 5-FC in a time dependent manner with induction of cytosine deaminase following infection of recombinant adenovirus. When 5-FC was removed 3 or 6 days after the treatment, the cells grew again. The effect was reproduced in the in vivo model of subconjunctival cellular proliferation although 5-FC was administrated as eye drops. There were no cases with corneal erosion. CONCLUSION: Cell proliferation was inhibited by co-exposure of 5-FC and cytosine deaminase. This new delivery system may merit controlled delivery of 5-FU after filtering surgery.

First Report of Anthracnose Caused by Colletotrichum gloeosporioides on Gueroba in Brazil.

Gueroba (Syagrus oleracea (Mart.) Becc.), a member of the family Arecaceae, is an important native palm tree in central Brazil and has great potential as a cultivated crop and ornamental plant (1). In July 1999, anthracnose symptoms were observed on several gueroba plants growing in a field in Planaltina, Federal District. Leaves showed spots or small circular to irregular, brown-to-black necrotic lesions, which generally coalesced as symptoms progressed. Tissue sections from leaves with symptoms were plated aseptically on potato dextrose agar (PDA). A fungus was consistently isolated from leaves of symptomatic plants and was identified as Colletotrichum gloeosporioides (Penzig) Penzig & Sacc. by P. F. Cannon of CABI Bioscience, Egham, UK, where a culture (IMI 384186) has been deposited. An isolate was tested for pathogenicity by inoculating 20 plants at the two-leaf stage by placing a mycelial PDA plug from a 10-day-old fungal culture on a previously wounded leaf. Noninoculated plants served as controls. Plants were enclosed in a plastic bag and incubated for 4 days at 26 ± 2°C and ≈100% relative humidity. Ten days after inoculation, plants developed leaf lesions similar to those observed in the field. Symptoms did not develop on the control plants. C. gloeosporioides was reisolated from 95% of the inoculated leaves, completing Koch's postulates. The pathogenicity test was repeated with the same results. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose on gueroba in Brazil. Reference: (1) H. Lorenzi. Page 288 in: Árvores Brasileiras. Plantarum. Nova Odessa, Brazil 1998.

Lack of association of mutations of the bestrophin gene with age-related macular degeneration in non-familial Japanese patients.

BACKGROUND: Heterozygous mutations of the bestrophin gene are associated with Best macular dystrophy (BMD). The bestrophin gene is specifically expressed in the retinal pigment epithelium. BMD is a hereditary form of macular degeneration that may develop subretinal neovascularisation similar to the wet type of age-related macular degeneration (AMD). PURPOSE: To study whether mutations of the bestrophin gene occur in non-familial Japanese AMD patients. METHODS: A total of 85 non-familial AMD patients (average age 67.5 years; 71 male, 14 female) diagnosed by indocyanine green angiography were screened. Among them, 69 patients (81 %) were classified as having wet type AMD. Genomic DNA was purified from the total blood and used as the template for polymerase chain reaction (PCR). All the exons of bestrophin gene were amplified by PCR. Mutation analysis was performed by SSCP using the ABI Prism 310 Genetic Analyzer (Perkin Elmer). Nucleotide sequence was determined by direct sequencing of the PCR amplicons. As the control, 105 non-AMD patients (average age 62.0 years; 52 male, 53 female) were screened by the same method. RESULTS: Only one AMD patient had a specific polymorphism in exon 2, but no mutations leading to amino acid substitutions were found. In exon 2 and 3, two further polymorphisms were detected in all AMD patients as well as normal controls. CONCLUSION: No mutations were found in the bestrophin gene in nonfamilial Japanese patients with AMD or in normal controls.

Rat liver in vivo replicative DNA synthesis test for short-term prediction of nongenotoxic (Ames-negative) hepatocarcinogenicity: a collaborative study of the Nongenotoxic Carcinogen Study Group of Japan.

A collaborative study was conducted to evaluate whether a replicative DNA synthesis (RDS) test using the rat liver can detect nongenotoxic (Ames-negative) hepatocarcinogens with three or seven daily administrations at dose-levels effective in long-term bioassays. The assay methods were well-validated by the 14 participants. Of six compounds tested, carbon tetrachloride (50 and 100 mg/kg), clofibrate (125 and 250 mg/kg), diethylstilbestrol (0.125 and 0.25 mg/kg) and urethane (100 mg/kg) gave positive results, methyl carbamate (200 and 400 mg/kg) exerted equivocal effects, and D,L-ethionine (125 mg/kg) failed to elevate RDS. These findings suggest that the RDS test can detect many nongenotoxic rat hepatocarcinogens with short-term administration at dose-levels used in long-term bioassays.

ONO-4007, an antitumor lipid A analog, induces tumor necrosis factor-alpha production by human monocytes only under primed state: different effects of ONO-4007 and lipopolysaccharide on cytokine production.

ONO-4007 is a synthetic lipid A analog that exhibits strong antitumor activity in several animal models via intratumoral production of tumor necrosis factor (TNF). In the present study the cytokine-inducing effect of ONO-4007 was investigated in human monocytes that were freshly isolated or had been incubated for 3 days with granulocyte-macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor. ONO-4007 induced slight production of TNF-alpha, Interleukin (IL)-1 beta, IL-6 and IL-12 in fresh monocytes but strongly induced TNF-alpha production in GM-CSF-treated monocytes. Monocytes treated with macrophage colony-stimulating factor were also primed to produce TNF-alpha in response to ONO-4007. In the production of IL-1 beta, IL-6 and IL-12, GM-CSF did not show a priming effect. In contrast to ONO-4007, lipopolysaccharide (LPS) induced significant amounts of all these cytokines in fresh monocytes. In whole blood, ONO-4007 failed to induce TNF-alpha, whereas LPS and LA-15-PP (Escherichia coli-type lipid A) strongly induced TNF-alpha production. In the GM-CSF-treated monocytes, both elimination of serum from the culture medium and anti-CD14 antibody treatment attenuated LPS-induced TNF-alpha production but not ONO-4007-induced TNF-alpha production. This study shows that ONO-4007 activates human monocytes/ macrophages to release TNF-alpha only in a primed state and suggests that ONO-4007 would activate these cells via different pathways from LPS. These differences could mean that ONO-4007 has potent antitumor activity with lower toxicity than LPS.

Intratumoral tumor necrosis factor induction and tumor growth suppression by ONO-4007, a low-toxicity lipid A analog.

BACKGROUND: ONO-4007 is a lipid A analog with low toxicity. MATERIALS AND METHODS: The antitumor activity and tumor necrosis factor (TNF)-inducing activity of ONO-4007 were compared with those of lipopolysaccharide (LPS) in WKAH rats bearing KDH-8 hepatoma cells. RESULTS: Weekly injections of ONO-4007 (3 and 10 mg/kg i.v.) suppressed tumor growth, but LPS (0.01 and 0.1 mg/kg i.v.) did not. A single injection of ONO-4007 (3 and 10 mg/kg i.v.) into tumor-bearing rats induced higher levels of endogenous TNF production in tumor tissues than LPS (0.001, 0.01 and 0.1 mg/kg i.v.). Repeated injections of LPS caused a reduction of TNF production in tumor tissues, whereas the reduction by ONO-4007 was less remarkable than that by LPS. Intratumoral injections of anti-rat TNF-alpha antibody attenuated the antitumor effect of ONO-4007. CONCLUSION: The antitumor effect of ONO-4007 is more pronounced than that of LPS and the effect is mediated by TNF produced in tumor tissues.

Restoration of immune responses in tumor-bearing mice by ONO-4007, an antitumor lipid A derivative.

ONO-4007, a synthetic lipid A derivative, has been found to exhibit potent antitumor activity in several animal models. In the present study, we examined the effects of ONO-4007 on delayed-type hypersensitivity (DTH) reaction and antibody production in Meth A sarcoma-bearing BALB/c mice. The DTH reaction to sheep red blood cells (SRBC) and the IgG production against keyhole limpet hemocyanin (KLH), were depressed in tumor-bearing mice as well as in normal mice given Mitomycin C (MMC). However, ONO-4007 restored these immune responses to normal levels. In addition, in vitro studies showed that ONO-4007 induced the production of tumor necrosis factor-alpha (TNF-alpha) in splenic adherent cells of tumor-bearing mice more than those in normal mice. Though ONO-4007 alone had little effect on the induction of IL-2 production in spleen cells, it augmented the Concanavalin A (Con A)-stimulated IL-2 production. Moreover, ONO-4007 had a mitogenic effect on spleen cells. These results suggest that ONO-4007 may improve immunocompetence in tumor-bearing hosts and contribute to the induction of antitumor immunity and prevention of bacterial infections.

[Reproductive pharmacology of prostaglandins].

Effects of prostaglandins and their analogues in reproduction were investigated using rats and Japanese monkeys. Prostaglandin of the F series exerted uterine contractile, luteolytic, anti-nidation and abortifacient effects. Prostaglandins of the E series exhibited a potent uterine contractile effect, although their luteolytic, anti-nidation and abortifacient effects were much less potent than those of the PGF series. A comparison of animal and human data on prostaglandins and their analogues indicated a good cor-relationship between the uterine contractile effect in animals (rates and monkeys) and the abortifacient or labor inducing effect in humans.

[Effect of OU-1308 on uterine motility in pregnant rats and monkeys].

Effect of OU-1308 (17s, 20-dimethyl-6-oxo-prostaglandin E1 methyl ester) on uterine motility in anesthetized rats and monkeys was examined by means of the balloon catheter or open-end catheter method and compared with that of PGF2 alpha. OU-1308 and PGE2 alpha exhibited uterine contractile activity at the dose of 30 micrograms/kg, i.v., on day 8 of pregnancy and 10 micrograms/kg, i.v., on day 20 of pregnancy in rats. In monkeys on day 50 approximately 120 of pregnancy, both compounds enhanced uterine motility at 10 micrograms/kg, i.v. Intragastric administration of OU-1308 at 500 micrograms/kg, however, was without effect in monkeys. These results indicate that when administered intravenously, OU-1308 was as potent as PGF2 alpha in terms of uterine contractile activity in pregnant rats and monkeys.

[Effects of 16,16-dimethyl-trans-delta 2-PGE1 methyl ester (ONO-802) on uterine motility (author's transl)].

In rats on day 20 of pregnancy, ONO-802 exerted a uterine contractile effect when 0.2 microgram/kg was given i.v. and also with an intrauterine administration of 0.5 ng, s.c. administration of 1 microgram/kg and vaginal administration of 10 microgram/kg. The contractile activity of ONO-802 was 100-1000 times more potent than that of PGE2 alpha and was long-acting. Intravenous PGE1 or PGE2 induced a transient contraction followed by inhibition of spontaneous uterine motility in nonpregnant rats and in those with an early pregnancy. In nonpregnant hamsters, a relaxation was seen. ONO-802, similar to PGF2 alpha, showed a contractile effect regardless of states of nonpregnancy and pregnancy, both rats and hamsters. IN rats, a contractile threshold dose of ONO-802 was similar from day 15 to day 21 of pregnancy. Oxytocin produced an acute decrease in this threshold dose on phentolamine or methysergide, but was inhibited by papaverine, dibutyryl cAMP, salbutamol and verapamil. These results suggest that uterine contraction induced by ONO-802 differs from the contractions induced by PGE1, PGE2, and oxytocin, that ONO-802 does not induce endogenous PGs, and that the effect of ONO-802 is not mediated by the autonomic nervous system.

[Effects of 16,16-Dimethyl-trans-delta 2-PGE1 methyl ester (ONO-802) on uterine, placental and ovarian circulation (author's transl)].

Doses of the drugs which produce uterine contraction were given intravenously to various species of animals. Placental and ovarian circulations were measured by the thermocouple method and/or microangiography. Uterine arterial blood flow (UABF) was measured by the electromagnetic flowmeter. Both placental blood flow (PBF) and UABF decreased with the administration of ONO-802, PGE1 or PGF2 alpha to to rabbits and dogs. Oxytocin and noradrenaline also decreased PBF in rabbits. ONO-802 or PGE1 lowered arterial blood pressure (BP) in rats, rabbits and dogs. PGE2 alpha elevated BP in rats and dogs and lowered it in rabbits. Oxytocin produced no changes in PB while noradrenaline elevated BP in rabbits. Ovarian blood flow in pregnant rabbits was reduced by ONO-802, PGE1 or PGF2 alpha. Little influence was seen with oxytocin. Regarding the luteal microvasculature in pregnant rats, ONO-802, PGF2 alpha and noradrenaline exhibited vasoconstricting effects. PGF2 alpha induced abortion and decreased plasma progesterone levels. These results suggest that the inhibitory effects of ONO-802 on the uterine and placental circulation are strongly influenced by the uterine contractile effect and that the inhibitory effect on the ovarian circulation in rats is one of pharmacological effects not concerned with the abortifacient or luteolytic effect.