A chimeric recombinant crustacean hyperglycemic hormone from Litopenaeus schmitti (Burkenroad) obtained as C‐terminus fusion protein boost hemolymph glucose concentration in Litopenaeus vannamei (Boone)

To date, no hormonal treatments are available for control of shrimp reproduction and only eyestalk ablation is of practical use. The crustacean hyperglycemic hormone (CHH) is the most abundant neuropeptide of the eyestalk CHH family. It plays an important role in the regulation of hemolymph glucose levels, as its principal function, but it is also implicated in additional physiological processes such as moulting and reproduction. In the present study, the cDNA encoding Litopenaeus schmitti (Burkenroad) mature CHH was cloned into the Escherichia coli pTYB2 expression vector. Using this strategy we have obtained, for the first time, the recombinant CHH from L. schmitti with its C‐terminus fused to an intein tag. The expected fused protein of about 63 KDa was expressed in E. coli forming inclusion bodies. It was purified in a soluble form by electroelution following molecular size fractionation in sodium dodecil sulphate polyacrylamide gel electrophoresis. The ability of the chimeric CHH protein to elevate glucose levels in the hemolymph of the eyestalk‐ablated Litopenaeus vannamei (Boone) shrimps indicates that its biological activity as hyperglycemic protein is preserved. The results provide an alternative tool to obtain soluble recombinant proteins from the CHH family of neuropeptides to get a better understanding of shrimp endocrinology.     

Problems in Paradise: Taiwanese Immigrants to Auckland,New Zealand

Migration from Asia to New Zealand is now a topic of heated public debate within New Zealand. This study traces the experiences of Taiwanese migrants to Auckland over the past decade and documents their adaption and revised expectations. By the end of June 1995, over 21,000 Taiwanese had been granted permanent residence in New Zealand on the basis of their skills, qualifications and work experience. Although expecting a fall in income as the necessary ‘price’ to pay for an improved quality of life, as well as some language difficulties, few immigrants were prepared for the magnitude of the drop in income, the hardship it would bring and the heavy reliance this would place on their remaining investments in Taiwan. The net result has been unemployment and underemployment of immigrants many of whom are now resigned to net gains through education coming not to them but to their children. As a result of the inability to secure adequate employment, many Taiwanese immigrants have relocated one or all of the family members back to Taiwan. The introduction in October 1995 of stricter English standards and the flatter, less academic basis to point earning qualifications, validation of job offers, recognition of New Zealand work experience together with the spouses’ human capital as well as a clearer commitment to New Zealand should all go some way to addressing many of the difficulties the history of recent Taiwanese immigration has uncovered.     

Induction of spawning in Pacific white shrimp Litopenaeus vannamei (Boone, 1931) by injection of its molt inhibiting hormone isoform II produced in E. coli

The optimization of reproductive parameters in shrimp farming continues to be a challenge for most producing countries. Although the crustacean neuropeptides have been studied extensively in the last two decades, the functions of most of these neuropeptides remained putative. Among them, molt‐inhibiting hormone isoform II (MIH II) has shown an important role in vitellogenesis. In this study, the cDNA encoding mature MIH II peptide was isolated by RT‐PCR from the L. vannamei eyestalk. The cDNA was cloned into pET28a bacterial expression vector. Recombinant MIH II was obtained in the form of insoluble inclusion bodies and purified to ~88% purity. Two doses of rMIH II and a negative control group were assayed in vivo. The stages of ovarian maturation and spawning were recorded during 72 hr post‐injection. The results showed that ovarian maturation occurred approximately in 9% and 33% of females injected with rMIH II at the doses of 300 and 600 ng/gbw respectively. Neither maturation nor spawning was detected in the negative control group. Females injected with 600 ng/gbw, which showed vitellogenic stages III and IV, spawned. These preliminary results argue that the hormone rMIH II could be a promising candidate to induce spawning in L. vannamei shrimp.     

Evaluation of Different Inoculation Methods for Screening of Sclerotinia rot and Phytophthora blight in Perilla Germplasm

Journal of Crop Science and Biotechnology - Sclerotinia rot and Phytophthora blight are devastating diseases of perilla that can cause huge loss of perilla production. Screening disease resistant...     

Global mapping of the yeast genetic interaction network

A genetic interaction network containing approximately 1000 genes and approximately 4000 interactions was mapped by crossing mutations in 132 different query genes into a set of approximately 4700 viable gene yeast deletion mutants and scoring the double mutant progeny for fitness defects. Network connectivity was predictive of function because interactions often occurred among functionally related genes, and similar patterns of interactions tended to identify components of the same pathway. The genetic network exhibited dense local neighborhoods; therefore, the position of a gene on a partially mapped network is predictive of other genetic interactions. Because digenic interactions are common in yeast, similar networks may underlie the complex genetics associated with inherited phenotypes in other organisms.     

Whole-genome shotgun assembly and analysis of the genome of Fugu rubripes

The compact genome of Fugu rubripes has been sequenced to over 95% coverage, and more than 80% of the assembly is in multigene-sized scaffolds. In this 365-megabase vertebrate genome, repetitive DNA accounts for less than one-sixth of the sequence, and gene loci occupy about one-third of the genome. As with the human genome, gene loci are not evenly distributed, but are clustered into sparse and dense regions. Some "giant" genes were observed that had average coding sequence sizes but were spread over genomic lengths significantly larger than those of their human orthologs. Although three-quarters of predicted human proteins have a strong match to Fugu, approximately a quarter of the human proteins had highly diverged from or had no pufferfish homologs, highlighting the extent of protein evolution in the 450 million years since teleosts and mammals diverged. Conserved linkages between Fugu and human genes indicate the preservation of chromosomal segments from the common vertebrate ancestor, but with considerable scrambling of gene order.     

Functional targeting of DNA damage to a nuclear pore-associated SUMO-dependent ubiquitin ligase

Recent findings suggest important roles for nuclear organization in gene expression. In contrast, little is known about how nuclear organization contributes to genome stability. Epistasis analysis (E-MAP) using DNA repair factors in yeast indicated a functional relationship between a nuclear pore subcomplex and Slx5/Slx8, a small ubiquitin-like modifier (SUMO)-dependent ubiquitin ligase, which we show physically interact. Real-time imaging and chromatin immunoprecipitation confirmed stable recruitment of damaged DNA to nuclear pores. Relocation required the Nup84 complex and Mec1/Tel1 kinases. Spontaneous gene conversion can be enhanced in a Slx8- and Nup84-dependent manner by tethering donor sites at the nuclear periphery. This suggests that strand breaks are shunted to nuclear pores for a repair pathway controlled by a conserved SUMO-dependent E3 ligase.     

Stability of hydrolytic enzyme activity and microbial phosphorus during storage of tropical rain forest soils

Storage can markedly influence microbial and biochemical properties in soils, yet recommendations for sample storage are based on studies of temperate soils that regularly experience extended cold periods. We assessed the influence of storage conditions on microbial phosphorus and the activity of four hydrolytic enzymes (phosphomonoesterase, phosphodiesterase, β-glucosidase, and N-acetyl-β-d-glucosaminidase) in three lowland tropical forest soils from the Republic of Panama that experience a constant warm temperature. The soils spanned a strong rainfall gradient and contained contrasting physical and chemical properties (pH 3.6-5.9; total carbon 26-50 g C kg⁻¹; clay 33-62%; total phosphorus 0.30-0.60 g P kg⁻¹). Storage treatments were: (i) room temperature (22 °C in the dark), (ii) refrigerated (4 °C in the dark), (iii) air-dried (10 d, 22 °C), and (iv) frozen (−35 °C). There were significant changes in enzyme activities and microbial phosphorus during refrigerated and room temperature storage, although changes were relatively small during the first two weeks. An initial marked decline in enzyme activities for one soil analyzed within 2 h of sampling was attributed to a flush of activity caused by sampling and soil preparation (sieving, etc.). For longer-term storage (>2 weeks), ambient laboratory temperature appeared preferable to freezing and cold storage, because one month of storage caused a marked decline in enzyme activities and microbial phosphorus in one soil. Freezing preserved the activities of some enzymes in some soils at rates comparable to cold or room temperature storage, but caused a marked decline in microbial phosphorus in two soils. Air-drying caused a marked decline in microbial phosphorus and the activity of all enzymes. We therefore conclude that enzyme assays and microbial phosphorus should be determined in tropical forest soils after no more than two weeks storage in the dark at ambient laboratory temperature.     

Nematode community composition and diversity associated with a topoclimatic transect in a rain shadow desert

The effect of a topoclimatic gradient on soil nematode communities was investigated in the Israeli Judean Desert. Four locations along a 35 km gradient, from an elevation of 650 m above sea level with 620 mm rainfall to a –60 m relative to sea level with a rainfall below 110 m, were studied: Givat Yearim, Maale Adumim, Mishor Adumim and Kalia. Monthly soil samples were collected between January 1994 and December 1995. Thirteen nematode families and 17 genera were observed. Cephalobus, Heterocephalobus, Aphelenchoides, Tylenchus, Dorylaimus and Eudorylaimus were found to be the dominant genera. Ecological measurements of soil nematode community structure, diversity, and maturity indices were assessed, and comparisons between four locations and four seasons were made. Significant differences in the total numbers of nematodes were found between the locations (p < 0.01) and seasons (p < 0.05), where Givat Yearim > Maale Adumim > Mishor Adumim > Kalia, winter > spring > autumn > summer. Bacterivores were found to be the most abundant trophic group across locations and seasons, with a mean relative abundance of 55.0%. The densities of bacterivores, plant parasites and omnivores-predators during the winter season exhibited similar trends at all four locations, with Givat Yearim > Maale Adumim > Mishor Adumim > Kalia. None of the ecological indices were significantly different between locations. However, the maturity index (MI), trophic diversity (TD) and Simpson diversity (SI) exhibited significant differences between seasons.     

Doppler sonography of uterine blood flow at mid‐oestrus during different degree of clinical endometritis in dairy cows

The present study was aimed at analysing the mid‐oestrous uterine blood flow parameters during varying degree of endometritis in dairy cows. Degree of clinical endometritis was adjudged on the basis of visual examination of uterine discharge collected from cows at mid‐oestrus, that is mild (slightly turbid; n = 13), moderate (turbid with pus flakes; n = 14) and severe (milky; n = 13). Pulsatility and resistance indices (PI and RI), time average mean velocity (TAMEAN), time average maximum velocity (TAMAX), diameter of the artery, volume of blood flow, Doppler pulse duration (DPD) and systolic upstroke/acceleration time (AT) were measured to study the spectral waveforms at mid‐oestrus. Significantly higher (p < .01) PI and RI were found in cows diagnosed with mild degree in comparison with cows diagnosed with moderate and severe degrees of clinical endometritis. There was significantly higher (p < .01) velocity (TAMEAN and TAMAX), volume of blood flow (BFV‐TAMEAN, BFV‐TAMAX) and DPD in both the middle uterine arteries during moderate and severe degrees of clinical endometritis as compared to mild endometritis. However, significantly higher (p < .01) AT was recorded in cows diagnosed with mild degree as compared to moderate and severe degrees of clinical endometritis. Pearson's correlation analysis has shown that RI was positively correlated with PI and AT in all the groups under study (mild degree, r = .72 and .49; moderate degree, r = .54 and .38; severe degree, r = .90 and .42; p < .05). However, there found significantly negative correlation (p < .05) with other parameters in all the cows irrespective of degree of inflammation. Therefore, it may be deducted that assessment of uterine inflammation can be done with a non‐invasive technique known as Doppler sonography which can be useful in adjudging the hemodynamic changes inside uterus and future fertility of dairy cows.