Examination of various biomarkers measuring genotoxic endpoints from Barcelona airport personnel

Three different biomarkers: sister-chromatid exchanges (SCE), micronuclei (MN), and the Comet assay, were used to evaluate different kinds of genetic damage in peripheral blood lymphocytes from 34 male workers at Barcelona airport, exposed to low levels of hydrocarbons and jet fuel derivatives. The control group consisted of 11 unexposed men. We also investigated the ras p21 protein levels in plasma, in order to evaluate whether the ras gene could serve as a suitable potential marker of carcinogenic pollution in occupationally exposed cohorts. SCE and MN analyses failed to detect any statistically significant increase in the airport workers when compared with the controls, and in fact, the frequency of binucleated cells with MN in the exposed group was significantly lower than that obtained in the control. However, slight but significant differences in the mean comet length and genetic damage index were observed between the exposed and control groups when using the Comet assay. There were no statistically significant differences between both groups in p21 plasma levels. Smoking was shown to affect significantly both SCE and high frequency cells (HFC) in the exposed group.     

Cytogenetic study of workers exposed to chromium compounds

The frequency of sister chromatid exchanges (SCEs), high SCE frequency cells (HFCs), and genetic polymorphism of genotypes glutathione S-transferase (GST) M1 and T1 were analyzed in peripheral lymphocytes of 35 workers occupationally exposed to chromium (Cr) and 35 matched control group. Results showed that workers exposed to Cr showed 6.07 SCE/cell, as compared to 4.76 SCE/cell for the control group (p<0.01). Smokers showed a statistically significant higher frequency of SCE than non-smokers in both groups. The work duration of Cr workers was an important factor. Workers exposed for more than 5 years showed a significantly higher level of SCEs (p<0.05). Workers exposed to Cr for 5 or more years had higher HFC rates (51.4%) than those exposed for less than 5 years (22.9%), with an odds ratio of 4.5 times than those exposed for less than 5 years. In HFC analysis, Cr workers who smoked showed a higher level of HFC (60%) than the control group (5.7%) and also had a higher odds ratio (60.4) compared with the control group. Among non-smokers, the odds ratio was 9.0. Another objective of this study is to investigate the relationship between SCE and genetic polymorphisms of GST M1 and T1 in Cr workers. The results showed that the incidence of GSTM1 null genotype was 60% in the control group and 77.1% in Cr workers, and percentages of GSTT1 deletion were 42.9% and 62.9% in control and exposed individuals, respectively. There was a slightly increased frequency of SCE among Cr workers with GSTM1 null genotype as opposed to non-null genotype individuals. A similar result was seen among the control group; however, there were no statistically significant differences. In conclusion, the current study found the positive induction of SCE in workers who smoked or/and were exposed to Cr. However, different GST genotypes did not influence the level of cytogenetic damage between groups. Despite slight variation in numbers, they all appear to be not different.     

Genetic and pathogenic diversity of Neofusicoccum parvum in New Zealand vineyards

Genetic diversity of 50 isolates of Neofusicoccum parvum, the predominant species of the Botryosphaeriaceae recovered from grapevines displaying symptoms of dieback and decline in New Zealand, was compared to that of isolates from Australia, South Africa, and California. The eight universally primed polymerase chain reaction (UP-PCR) primers distinguished 56 genotypes, with only four clonal pairs found. Seven main groups were identified in a neighbour-joining (NJ) tree with isolates from different regions and vineyards of New Zealand, Australia, and California distributed in different groups, indicating a high level of intra and intervineyard genetic variation. All of the South African isolates were positioned in a separate UP-PCR group, indicating that these isolates were less related to the other N. parvum isolates. When compared to fungi that reproduce sexually the genetic diversity and Shannon diversity indices were low (0.076-0.249; 0.109-0.367, respectively), genetic identity levels were high (0.76-0.95), and genetic distance levels were low (0.04-0.27). The large number of genotypes and the low number of clones in the New Zealand N. parvum populations may be explained by parasexual recombination as anastomosis was observed between nonself pairings. Pathogenicity tests using isolates from different UP-PCR groups inoculated onto either green shoots or 1-y-old grapevines detected virulence diversity, indicating intra and intervineyard variation between isolates, however, no correlation was detected between UP-PCR group and virulence.     

Examination of various biomarkers measuring genotoxic endpoints from Barcelona airport personnel

Three different biomarkers: sister-chromatid exchanges (SCE), micronuclei (MN), and the Comet assay, were used to evaluate different kinds of genetic damage in peripheral blood lymphocytes from 34 male workers at Barcelona airport, exposed to low levels of hydrocarbons and jet fuel derivatives. The control group consisted of 11 unexposed men. We also investigated the ras p21 protein levels in plasma, in order to evaluate whether the ras gene could serve as a suitable potential marker of carcinogenic pollution in occupationally exposed cohorts. SCE and MN analyses failed to detect any statistically significant increase in the airport workers when compared with the controls, and in fact, the frequency of binucleated cells with MN in the exposed group was significantly lower than that obtained in the control. However, slight but significant differences in the mean comet length and genetic damage index were observed between the exposed and control groups when using the Comet assay. There were no statistically significant differences between both groups in p21 plasma levels. Smoking was shown to affect significantly both SCE and high frequency cells (HFC) in the exposed group.     

Elevated chromosome translocation frequencies in New Zealand nuclear test veterans

In 1957/58 the British Government conducted a series of nuclear tests in the mid-Pacific codenamed Operation Grapple, which involved several naval vessels from Britain and New Zealand. Two New Zealand frigates with 551 personnel onboard were stationed at various distances between 20 and 150 nautical miles from ground zero. In the present study we applied the cytomolecular technique mFISH (multicolour fluorescent in situ hybridisation) to investigate a potential link between chromosome abnormalities and possible past radiation exposure in New Zealand nuclear test veterans who participated in Operation Grapple. Compared to age matched controls, the veterans showed significantly higher (P < 0.0001) frequencies of chromosomal abnormalities (275 translocations and 12 dicentrics in 9,360 cells vs. 96 translocations and 1 dicentric in 9,548 cells in the controls), in addition to a significant excess of CCRs (complex chromosomal rearrangements) in the veterans. A Kolmogorov-Smirnoff test showed that the distributions of translocations for the two groups were significantly different.     

Cytogenetic analysis in workers occupationally exposed to nickel carbonyl

Chromosomal aberration and sister-chromatid exchange (SCE) base-line frequencies and SCE frequencies induced by 10 ng/ml mitomycin C (MMC) were analysed in cultured peripheral lymphocytes of 65 workers occupationally exposed to nickel carbonyl Ni(CO)4. The subjects were divided into 4 groups: (1) control; (2) exposed to nickel carbonyl (= exposed); (3) cigarette smokers; (4) smoking-exposed. The results show that there are no significant differences in chromosomal aberration frequencies, breaks or gaps, between the various groups. However, the SCE base-line frequency of the smoking-exposed group, with an average of 7.7/cell, was significantly higher than that of the control group, with an average of 6.5/cell (P less than 0.01), and also than that of the exposed group with an average of 5.9/cell (P less than 0.01). Similarly, the SCE frequency induced by 10 ng/ml MMC in the smoking-exposed group which averaged 15.5/cell was significantly higher than that of the control group (average of 13.2/cell (P less than 0.05], and also than that of the exposed group with an average of 12.3/cell (P less than 0.01). Under our experimental conditions, it may be that the level of exposure was not high enough to elicit an increase in chromosomal aberrations and SCE frequencies in the non-smoker exposed group. The fact that an increase in SCE frequencies was only found in the smoking-exposed group implies that the two factors, smoking and exposure to nickel carbonyl, are jointly responsible for the result.     

Genetic structure of an introduced paper wasp,Polistes chinensis antennalis (Hymenoptera,Vespidae) in New Zealand

Several eusocial wasps are prominent invaders to remote islands. The paper wasp Polistes chinensis antennalis is native to East Asia, was introduced to New Zealand in 1979 and has expanded its distribution there. This provides an excellent opportunity to examine the impacts of an initial bottleneck and subsequent expansion on genetic structure. We analysed and compared the genetic population structures of the native (Japan and South Korea) and invasive New Zealand populations. Although 94% of individuals had shared haplotypes detected across both populations, the remaining 6% had private haplotypes identified in only one of the three countries. The genetic variation at microsatellite loci was lower in New Zealand than in native countries, and the genetic structure in New Zealand was clearly distinct from that in its native range. Higher frequencies of diploid‐male‐ and triploid‐female‐producing colonies were detected in New Zealand than in the native countries, showing the reduction in genetic variation via a genetic bottleneck. At least two independent introductions were suggested, and the putative source regions for New Zealand were assigned as Kanto (central island) and Kyushu (south island) in Japan. Serial founder events following the initial introduction were also indicated. The estimated dispersal distance between mother and daughter in New Zealand was twice that in Japan. Thus, the introduction history of P. chinensis antennalis in New Zealand is probably the result of at least two independent introductions, passing through a bottleneck during introduction, followed by population expansion from the point of introduction.     

SCE increases after an accidental acute inhalation exposure to EtO and recovery to normal after 2 years

Sister-chromatid exchanges (SCEs) were studied in 3 workers accidentally exposed for about half an hour to high levels of ethylene oxide (more than 700 ppm) during a sterilizing process. The 3 workers had clinical symptoms of exposure and were tested for SCE frequencies 5 days and 2 years after the accident: 2 had also been tested 6 months earlier. All 3 showed a similar increase in SCEs after the accident, to a mean of 13.8 SCEs/cell compared with 8.6 for a control group. The incidence of 'high-frequency cells' was markedly elevated but decreased over 2 years, when the mean SCE frequencies had also returned to pre-accident levels.     

Long term depleted uranium exposure in Gulf War I veterans does not cause elevated numbers of micronuclei in peripheral blood lymphocytes

Depleted uranium (DU) is a high density heavy metal that has been used in military munitions since the 1991 Gulf War. DU is weakly radioactive and chemically toxic. Long term exposure can cause adverse health effects. This study assessed genotoxic effects in DU exposed Gulf War I veterans as a function of uranium (U) body burden. Levels of urine U were used to categorize the cohort into low and high exposure groups. Exposure to DU occurred during friendly fire incidents in 1991 involving DU munitions resulting in inhalation and ingestion exposure to small particles of DU and soft tissue DU fragments from traumatic injuries. All of these Veterans are enrolled in a long term health surveillance program at the Baltimore Veterans Administration Medical Center. Blood was drawn from 35 exposed male veterans aged 36-59 years, then cultured and evaluated for micronuclei (MN) using the cytokinesis block method. The participants were divided into two exposure groups, low and high, based on their mean urine uranium (uU) concentrations. Poisson regression analyses with mean urine U concentrations, current smoking, X-rays in the past year and donor age as dependent variables revealed no significant relationships with MN frequencies. Our results indicate that on-going systemic exposure to DU occurring in Gulf War I Veterans with DU embedded fragments does not induce significant increases in MN in peripheral blood lymphocytes compared to MN frequencies in Veterans with normal U body burdens.     

Genetic differentiation of starling (Sturnus vulgaris: Aves) populations in New Zealand and Great Britain

Several hundred starlings (Sturnus vulgaris) were introduced to New Zealand from Great Britain during1860–1880. Allozymic variation at 24 loci was analysed in winter populations sampled at six localities in each country. New Zealand samples had fewer alleles per locus but the same mean heterozygosity (3% per locus) and proportion of polymorphic loci as did British samples. Winter populations in Britain contain European migrants and were genetically homogeneous. Paradoxically, genetic distances among derived New Zealand populations, and between New Zealand and Great Britain were much greater, similar in magnitude to those observed among allopatric populations in other avian species. The geographical pattern of genetic variation in New Zealand suggests that reproductive isolation of populations and random drift have contributed to the development of population differentiation.     

Metabolite ratio of toluene-exposed rotogravure printing plant workers reflects individual mutagenic risk by sister chromatid exchanges

The study involved a group of 42 printing plant workers and a control group of 45 blood donors. At the working places, the ambient air-toluene concentration amounted from 141 to 328 mg/m(3). Sister chromatid exchanges (SCEs) were significantly elevated by three units in the exposed group. In this group, the concentration of urinary toluene metabolites was also considerably increased-hippuric acid was four times higher and the o-cresol and p-cresol fractions were twice as high. Results of toluene monitoring of ambient air- or blood-toluene concentrations did not show any relationships with individual SCE. While these SCE values revealed only a weak relationship with the corresponding hippuric acid data, a significant correlation with the cresols, which are known to be more genotoxic than hippuric acid, appeared in highly exposed workers. An attempt was made to consider the individual metabolic balance of toluene excretion products. For that reason individual cresol to hippuric acid ratios were calculated and related to corresponding SCE values. In all investigated subpopulations of the exposed group, this ratio correlated with SCE at a level of high significance. This strong interrelationship is a powerful argument for the genotoxic behavior of toluene. Furthermore, the individual metabolic balance, as a consequence of genetic polymorphism, should be considered in the discussion about genetic risk of toluene.     

二磷酸盐促进假体周围骨溶解后假体翻修骨整合的实验研究

目的:评价阿仑膦酸钠对新西兰大白兔假体周围发生骨溶解后,再进行对新西兰大白兔假体翻修骨整合的影响。方法:选取雄性新西兰大白兔30只,随机分成3组(正常组,实验组,对照组,每组10只)。正常组在胫骨松质骨区域植入钛合金假体,实验组和对照组分别植入钛合金假体和钛颗粒,饲养8周后,三组统一进行假体翻修。实验组用阿仑磷酸钠治疗8周后取材,对照组和正常翻修组也分别进行取材。通过假体推出力学实验、硬组织切片观察,评价阿仑磷酸钠对假体周围翻修后骨整合的影响。结果:假体推出力学实验结果显示,实验组假体最大推出载荷明显大于对照组(P0.01)。硬组织学切片通过苦味酸--品红染色,利用图像分析仪器统计假体周围骨整合的面积实验组假体周围骨量明显优于对照组假体周围骨量(P0.05)。结论:二磷酸盐-阿仑磷酸钠可以提高假体翻修后假体周围骨整合。     

Sister chromatid exchanges in patients with precancerous and cancerous lesions of cervix uteri

Summary The frequency of sister chromatid exchanges (SCEs) was studied in leucocytes from 46 patients with cervical carcinoma, 89 precancerous lesions, and 43 age-matched control women. The frequency of SCEs was found to be 10.15 ±2.49 in cancer, 8.83±2.15 in precancerous lesions, and 7.55±2.24 in controls. The analyses of SCE data revealed a highly significant (P<0.001) increase in precancerous and cancerous lesions compared to controls. The intra-chromosomal distribution of SCEs revealed a random increase in various chromosomal groups in patients with cancer and dysplasia compared to controls. The mean SCE level among various groups of precancerous lesions according to severity of pathological condition did not show significant differences. However, 70.8% of dysplasia cases revealed SCE levels higher than the average in controls. The increased frequencies of SCEs in the majority of cancer patients and a few, precancerous lesions indicate that individuals with high SCE levels may be at a high risk of developing cancer. Thus the usefulness of SCE levels as a preclinical marker to identify the high risk group of dysplasias needs to be ascertained by follow-up studies; these are in progress.     

Sister-chromatid exchanges in association with occupational exposure to ethylene oxide

Sister-chromatid exchange (SCE) frequencies in employees potentially exposed to ethylene oxide (ETO) were compared with those in unexposed control groups. Three worksites where the previous environmental control of ETO was known to have differed were chosen. Within these worksites, subjects were categorized into high potential exposed, low potential exposed and control groups. An additional community control group was obtained. Blood samples for chromosome studies of peripheral lymphocytes were drawn at several time points over a period of 24 months. The effects on SCE of age, sex, smoking habits and reader variation were considered. Worksites I, II and III, respectively, represented increasing levels of exposure. At Worksite III large differences among groups persisted over 24 months. At Worksite II, the SCEs in the high potential exposed workers were higher than those in the other groups. At no time was the low potential exposed group at Worksite II statistically significantly higher in mean SCE than the worksite controls. No consistent differences among groups were noted in Worksite I.     

Invasion success of the bumblebee, Bombus terrestris, despite a drastic genetic bottleneck

In early 1992, the European bumblebee, Bombus terrestris, was first seen in Tasmania and currently has spread to most of the island. Here, we report on the genetic structure, using micro-satellites, of the invading population from samples collected in the years 1998-2000, a few years after the first sighting of the species in its new area. The data show that the Tasmanian population has a very low genetic diversity, with less than half of the allelic richness (Richness=2.89 alleles; H(exp)=0.591) and lower levels of heterozygosity as compared to populations in New Zealand (4.24 alleles; H(exp)=0.729) and Europe (5.08 alleles; H(exp)=0.826). In addition, the genetic data suggest that the invasion must have happened once, probably around late 1991, and was the result of very few, perhaps only two, individuals arriving in Tasmania. Furthermore, these founders came from the New Zealand population. Today, the population in the south of Tasmania seems to act as a source population from which individuals migrate into other parts of the state. A similar source-sink structure seems also the case for New Zealand. The data show that B. terrestris is a highly invasive species capable of establishing itself even after a dramatic genetic bottleneck. B. terrestris may be an invasive species due to the haplo-diploid sex determination system, which exposes recessive, deleterious mutations to selection. Offspring of such purged lines may then be able to tolerate high levels of inbreeding.     

Few genetic differences between Victorian and Western Australian blue penguins,Eudyptula minor

Abstract

Blue penguins, Eudyptula minor, breeding on Penguin Island, Western Australia are considerably larger than other blue penguins in Australia. If genetic isolation is the cause, it may have implications for the conservation status of some blue penguin populations. We compared the sequences of two mitochondrial gene regions (cytochrome‐b and the control region) from Western Australian blue penguins with other populations of blue penguins from Australia and New Zealand. We found few differences between sequences from Western Australia, Phillip Island, Victoria and Otago, New Zealand, although all three differed considerably from other New Zealand blue penguins. Sequences for the control region from the Western Australian blue penguins and 30 more birds breeding at various Australasian sites provided further support for two major clades within Eudyptula; an Australian clade (including Otago) and a New Zealand clade.     

Population genetic structure and taxonomy of the common dolphin (Delphinus sp.) at its southernmost range limit: New Zealand waters

New Zealand is the southernmost limit of the common dolphin's (genus Delphinus) distribution in the Pacific Ocean. In this area, common dolphins occur in both coastal and oceanic habitats, exhibit seasonal and resident occurrence, and present high morphological variability. Here we investigated the population structure and the taxonomic identity of common dolphins (Delphinus sp.) within New Zealand waters using 14 microsatellite loci, 577 bp of the mtDNA control region, and 1,120 bp of the mtDNA cytochrome b gene across 90 individuals. We found high genetic variability and evidence of population expansion. Phylogenetic analyses conducted to clarify the taxonomic status of New Zealand common dolphins did not show any clustering reflecting geographic origin or morphotypes. The microsatellite analysis showed genetic differentiation between Coastal and Oceanic putative populations, while mtDNA revealed significant genetic differentiation only between the Hauraki Gulf and other putative groups. Our results suggest that differences in habitat choice and possible female site fidelity may play a role in shaping population structure of New Zealand common dolphins.     

Performance differences among ex situ native-provenance collections of Pinus radiata D. Don. 1: potential for infusion into breeding populations in Australia and New Zealand

Growth and form traits from a series of three provenance trials of Pinus radiata D. Don planted in New Zealand and Australia were analysed at age 9 years from planting. The trials included selections from three mainland California natural populations—A?o Nuevo, Monterey and Cambria. Monterey and Cambria performed better than A?o Nuevo at two New Zealand sites, but Monterey and A?o Nuevo were almost identical in growth, whereas Cambria grew less vigorously at the Australian site. We detected significant provenance differences for diameter at breast height (DBH) growth and stem straightness across countries (p < 0.001). Estimated heritability for DBH ranged from 0.19 to 0.26 within sites, while heritability estimates for stem straightness and branching frequency ranged from 0.10 to 0.24. Estimated type B genetic correlations for DBH were always higher between the two trials in New Zealand trials than between pairs of trials in New Zealand and the Australian site. The genetic coefficient of variation (CV_A) for DBH was around 8–10% compared to ca. 5% for the current breeding population. These results suggested that there is appreciable genetic variation in the native populations, and infusion of these materials would increase the genetic variation in current breeding populations. Ten unrelated parents ranked above control seedlots from the older open-pollinated seed orchard stock for DBH growth and would be potential candidates for infusion. The promising performance of the Cambria material is an important result because the genetic base of the present Australian and New Zealand plantations is principally derived from A?o Nuevo and Monterey.     

A novel locus regulates both retroviral glycoprotein 70 and anti-glycoprotein 70 antibody production in New Zealand mice when crossed with BALB/c

Lupus-prone New Zealand Black and New Zealand White mice produce high serum levels of the endogenous retroviral envelope protein gp70 and develop an Ab response to this autoantigen as part of their autoimmune disease. Linkage analysis of two crosses involving New Zealand and BALB/c mice mapped these traits to a group of overlapping loci, including a novel locus on proximal chromosome 12. This locus was linked with serum gp70 and the autoimmune response against it. The linkage of serum gp70 levels to a previously described locus on distal chromosome 4 was also confirmed. Sequence analysis of a candidate gene on distal chromosome 4, Fv1, provided support that this gene may be associated with the control of serum gp70 levels in both New Zealand Black and New Zealand White mice. Linkage data and statistical analysis confirmed a close correlation between gp70 Ag and anti-gp70 Ab levels, and together gave support to the concept that a threshold level of gp70 is required for the production of anti-gp70 Abs. Serum levels of anti-gp70 Abs were closely correlated with the presence of renal disease, more so than anti-dsDNA Abs. Understanding the genetic basis of this complex autoantigen-autoantibody system will provide insight into the pathogenesis of lupus in mice, which may have implications for human disease.