Cloning of a chub metallothionein cDNA and development of competitive RT-PCR of chub metallothionein mRNA as a potential biomarker of heavy metal exposure

Metallothionein has been assayed in a range of aquatic animal tissues as an indicator of metal exposure. We sequenced chub (Leuciscus cephalus) metallothionein cDNA which showed over 90% homology to common carp, goldfish and stone loach and 77% homology to rainbow trout sequences for metallothionein. We then used the extended primer method to develop an accurate quantitative competitive RT-PCR assay for metallothionein mRNA. RT-PCR was used to measure metallothionein mRNA in feral chub from a range of field sites, with different levels of heavy metal pollution, in the West Midlands, UK. Measurements were complemented by analysis of liver and gill metallothionein protein by capillary electrophoresis. There was no significant difference in the metallothionein protein levels between fish of different rivers and there was no evidence of elevation of mRNA at the sites of highest metal exposure. The level of metal exposure (e.g. zinc, nickel and cadmium each ranging between 15 and 28 microg/l ) at the pH (7.5-8.5) of these rivers appears insufficient to elevate hepatic or gill metallothionein in chub. A lack of elevation of hepatic metallothionein mRNA in chub exposed to zinc, copper and manganese for 24 h and 10 days in the laboratory also suggests a non-responsiveness of this species.     

用16S rRNA基因的内切酶图谱快速鉴别几种对虾病原菌

坎普氏弧菌是青岛海洋大学生物系微生物实验室于１９８９－１９９０年自对虾养殖场中国对虾红腿病心脏及血淋巴中分离并鉴定的菌株，副溶血菌和溶藻胶弧菌两菌株于１９９４年９月得自中国科学院微生物研究所，为建立快速、准确的中国对虾病原菌的诊断技术，根据几种细菌的１６ＳｒＲＮＡ基因的序列，设计并合成该基因的多聚酶反应的引物ＰＬ１和ＰＬ２。并用该对引物分别从坎普氏弧菌、副溶血弧菌和溶藻胶弧菌的ＤＮＡ要品中扩增出分     

Diagnosis of iridovirus in large yellow croaker by PCR

A rapid and sensitive PCR-based method for the detection of the large yellow croaker iridovirus(LYCIV) is described,which involves the amplification of a 295 bp fragment of the LYCIV ATPase gene from DNA isolated from naturally infected fish spleen.Sequencing of LYCIV ATPase gene fragment showed it shared 100% nucleotide sequence homology with the corresponding region of the ATPase gene of red sea bream iridovirus(RSIV) and sea bass iridovirus(SBIV),suggesting that LYCIV was homologous with RSIV and SBIV at least in part of the gemone.The specificity and sensitivity of the PCR procedure were tested on the iridovirus-infected fishes,the expected fragment was detected from spleen DNA samples of infected fishes,whereas no fragments were amplified from healthy fish spleen DNA,white spot syndrome baculoviruses(WSBV) DNA and pseudorabies virus(PRV) DNA.Detection limit of this method was 10^-7 ng positive plasmid DNA containing target sequence,equal to about 100 virions.In the infected experiment,first positive detection(1/4) appeared at Day 3 post-infection,all fish(4/4) tested positive at Day 7,however obvious symptoms were observed at Day 8,so LYCIV infection could be detected prior to the appearance of obvious symptoms.These results indicate that this PCR method could be used for early,rapid and specific detection of LYCIV infection.     

Three‐dimensional simulation of tsunami run‐up around conical island

At the circular Babi Island in the Flores tsunami (1992) and pear shaped island in the Okushiri event (1993), unexpectedly large tsunami run‐up heights in the lee of conic islands were observed. The flume and basin physical model studies were conducted in the Coastal Hydraulic Laboratory, Engineering Research and Development Center, U.S. Army Corps of Engineers to provide a better understanding of the physical phenomena and verify numerical models used in predicting tsunami wave run‐up on beaches, islands, and vertical walls. Reasonably accurate comparison of run‐up height of solitary waves on a circular island has been obtained between laboratory experimental results and two‐dimensional computation model results. In this study we apply three‐dimensional RANS model to simulate wave run‐up on conical island. In the run‐up computation we obtain that 3D calculations are in very good comparison with laboratory and 2D numerical results. A close examination of the three‐dimensional velocity distribution around conical island to compare with depth‐integrated model is performed. It is shown that the velocity distribution along the vertical coordinate is not uniform: and velocity field is weaker in the bottom layer and higher on the sea surface. The maximum difference (about 40%) appears at the time when solitary wave reached the circular island.     

Rare Earth Deposits of North America

Rare earth elements (REE) have been mined in North America since 1885, when placer monazite was produced in the southeast USA. Since the 1960s, however, most North American REE have come from a carbonatite deposit at Mountain Pass, California, and most of the world’s REE came from this source between 1965 and 1995. After 1998, Mountain Pass REE sales declined substantially due to competition from China and to environmental constraints. REE are presently not mined at Mountain Pass, and shipments were made from stockpiles in recent years. Chevron Mining, however, restarted extraction of selected REE at Mountain Pass in 2007. In 1987, Mountain Pass reserves were calculated at 29 Mt of ore with 8.9% rare earth oxide based on a 5% cut‐off grade. Current reserves are in excess of 20 Mt at similar grade. The ore mineral is bastnasite, and the ore has high light REE/heavy REE (LREE/HREE). The carbonatite is a moderately dipping, tabular 1.4‐Ga intrusive body associated with ultrapotassic alkaline plutons of similar age. The chemistry and ultrapotassic alkaline association of the Mountain Pass deposit suggest a different source than that of most other carbonatites. Elsewhere in the western USA, carbonatites have been proposed as possible REE sources. Large but low‐grade LREE resources are in carbonatite in Colorado and Wyoming. Carbonatite complexes in Canada contain only minor REE resources. Other types of hard‐rock REE deposits in the USA include small iron‐REE deposits in Missouri and New York, and vein deposits in Idaho. Phosphorite and fluorite deposits in the USA also contain minor REE resources. The most recently discovered REE deposit in North America is the Hoidas Lake vein deposit, Saskatchewan, a small but incompletely evaluated resource. Neogene North American placer monazite resources, both marine and continental, are small or in environmentally sensitive areas, and thus unlikely to be mined. Paleoplacer deposits also contain minor resources. Possible future uranium mining of Precambrian conglomerates in the Elliott Lake–Blind River district, Canada, could yield by‐product HREE and Y. REE deposits occur in peralkaline syenitic and granitic rocks in several places in North America. These deposits are typically enriched in HREE, Y, and Zr. Some also have associated Be, Nb, and Ta. The largest such deposits are at Thor Lake and Strange Lake in Canada. A eudialyte syenite deposit at Pajarito Mountain in New Mexico is also probably large, but of lower grade. Similar deposits occur at Kipawa Lake and Lackner Lake in Canada. Future uses of some REE commodities are expected to increase, and growth is likely for REE in new technologies. World reserves, however, are probably sufficient to meet international demand for most REE commodities well into the 21st century. Recent experience shows that Chinese producers are capable of large amounts of REE production, keeping prices low. Most refined REE prices are now at approximately 50% of the 1980s price levels, but there has been recent upward price movement for some REE compounds following Chinese restriction of exports. Because of its grade, size, and relatively simple metallurgy, the Mountain Pass deposit remains North America’s best source of LREE. The future of REE production at Mountain Pass is mostly dependent on REE price levels and on domestic REE marketing potential. The development of new REE deposits in North America is unlikely in the near future. Undeveloped deposits with the most potential are probably large, low‐grade deposits in peralkaline igneous rocks. Competition with established Chinese HREE and Y sources and a developing Australian deposit will be a factor.     

Zircon M257 ‐ a Homogeneous Natural Reference Material for the Ion Microprobe U‐Pb Analysis of Zircon

We introduce and propose zircon M257 as a future reference material for the determination of zircon U‐Pb ages by means of secondary ion mass spectrometry. This light brownish, flawless, cut gemstone specimen from Sri Lanka weighed 5.14 g (25.7 carats). Zircon M257 has TIMS‐determined, mean isotopic ratios (2s uncertainties) of 0.09100 ± 0.00003 for ²⁰⁶pb/²³⁸U and 0.7392 ± 0.0003 for ²⁰⁷pb/²³⁵U. Its ²⁰⁶pb/²³⁸U age is 561.3 ± 0.3 Ma (unweighted mean, uncertainty quoted at the 95% confidence level); the U‐Pb system is concordant within uncertainty of decay constants. Zircon M257 contains ～ 840 μg g^?1 U (Th/U ～ 0.27). The material exhibits remarkably low heterogeneity, with a virtual absence of any internal textures even in cathodoluminescence images. The uniform, moderate degree of radiation damage (estimated from the expansion of unit‐cell parameters, broadening of Raman spectral parameters and density) corresponds well, within the “Sri Lankan trends”, with actinide concentrations, U‐Pb age, and the calculated alpha fluence of 1.66 × 10¹⁸ g^?1. This, and a (U+Th)/He age of 419 ± 9 Ma (2s), enables us to exclude any unusual thermal history or heat treatment, which could potentially have affected the retention of radiogenic Pb. The oxygen isotope ratio of this zircon is 13.9%o VSMOW suggesting a metamorphic genesis in a marble or calc‐silicate skarn.     

Unequal Competition Among Chains of Supercenters: Kmart,Target, and Wal‐Mart

The three largest discount chains, Kmart, Target, and Wal‐Mart, have adopted the supercenter format for expansion. This article examines the locations of and the competition among these supercenters. Wal‐Mart has the greatest number of supercenters and is driving the expansion of the supercenter format. Both Kmart and Target have clustered their supercenters in a small number of metropolitan markets. In contrast to Wal‐Mart and Kmart, Target has focused on a middle‐class rather than a blue‐collar clientele. Now Kmart is struggling following a declaration of bankruptcy. Wal‐Mart has largely defeated its supercenter competitors and now is confronting the major grocery chains for grocery sales.     

Tests on low‐ductility RC frames under high‐ and low‐frequency excitations

The response of low‐ductility reinforced concrete (RC) frames, designed typically for a non‐seismic region, subjected to two frequencies of base excitations is studied. Five half‐scaled, two‐bay, two‐storey, RC frames, each approximately 5 m wide by 3.3 m high, were subjected to both horizontal and/or vertical base excitations with a frequency of 40 Hz as well as a lower frequency of about 4 Hz (close to the fundamental frequency) using a shake table. The imposed acceleration amplitude ranged from 0.2 to 1.2g. The test results showed that the response characteristics of the structures differed under high‐ and low‐frequency excitations. The frames were able to sustain high‐frequency excitations without damage but were inadequate for low‐frequency excitations, even though the frames exhibited some ductility. Linear‐elastic time‐history analysis can predict reasonably well the structural response under high‐frequency excitations. As the frames were not designed for seismic loads, the reinforcement detailing may not have been adequate, based on the crack pattern observed. The effect of vertical excitation can cause significant additional forces in the columns and moment reversals in the beams. The ‘strong‐column, weak‐beam’ approach for lateral load RC frame design is supported by experimental observations. Copyright © 2001 John Wiley & Sons, Ltd.     

Seismic response of self‐centring hysteretic SDOF systems

The seismic response of single‐degree‐of‐freedom (SDOF) systems incorporating flag‐shaped hysteretic structural behaviour, with self‐centring capability, is investigated numerically. For a SDOF system with a given initial period and strength level, the flag‐shaped hysteretic behaviour is fully defined by a post‐yielding stiffness parameter and an energy‐dissipation parameter. A comprehensive parametric study was conducted to determine the influence of these parameters on SDOF structural response, in terms of displacement ductility, absolute acceleration and absorbed energy. This parametric study was conducted using an ensemble of 20 historical earthquake records corresponding to ordinary ground motions having a probability of exceedence of 10% in 50 years, in California. The responses of the flag‐shaped hysteretic SDOF systems are compared against the responses of similar bilinear elasto‐plastic hysteretic SDOF systems. In this study the elasto‐plastic hysteretic SDOF systems are assigned parameters representative of steel moment resisting frames (MRFs) with post‐Northridge welded beam‐to‐column connections. In turn, the flag‐shaped hysteretic SDOF systems are representative of steel MRFs with newly proposed post‐tensioned energy‐dissipating connections. Building structures with initial periods ranging from 0.1 to 2.0s and having various strength levels are considered. It is shown that a flag‐shaped hysteretic SDOF system of equal or lesser strength can always be found to match or better the response of an elasto‐plastic hysteretic SDOF system in terms of displacement ductility and without incurring any residual drift from the seismic event. Copyright © 2002 John Wiley & Sons, Ltd.