Genetic differentiation of <Emphasis Type="Italic">Ganaspis brasiliensis</Emphasis> (Hymenoptera: Figitidae) from East and Southeast Asia

Ganaspis brasiliensis (Ihering) (Hymenoptera: Figitidae: Eucoilinae) is a Drosophila parasitoid that has often been misidentified as G. xanthopoda (Ashmead) in recent studies. This study aims to clarify genetic differentiation of G. brasiliensis based on the nucleotide sequences of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene and three nuclear DNA regions, the inter-transcribed spacers 1 and 2 (ITS1 and ITS2) and putative 60S ribosomal protein L37 (RpL37), as well as crossing experiments. Four lineages are recognized in individuals assigned as G. basiliensis by morphology, (1) individuals occurring in Japan and probably South Korea, (2) individuals from a small subtropical island of Japan, Iriomote-jima, (3) individuals from temperate lowlands of Japan and high altitude areas of Southeast Asia, and (4) individuals occurring widely in Asia, America, Hawaii and Africa. The first lineage is a specialist of Drosophila suzukii (Matsumura), a pest of fresh fruit, and also the fourth lineage has a capacity to parasitize this pest species. The first, third and fourth lineages occur sympatrically at least in Tokyo. The third and fourth lineages differed in mate choice and host use to some extent, but post-mating isolation between them was almost absent.     

Molecular cloning of a cDNA and assignment of the C-terminal of sarcotoxin IA, a potent antibacterial protein of Sarcophaga peregrina.

A previous paper described the complete amino acid sequences of sarcotoxins IA, IB and IC, which are a group of potent antibacterial proteins with almost identical primary structures produced by Sarcophaga peregrina (fleshfly) larvae [Okada & Natori (1985) J. Biol. Chem. 260, 7174-7177]. The present paper describes the cDNA cloning and complete nucleotide sequencing of a cDNA clone for sarcotoxin IA. The C-terminal amino acid residue of sarcotoxin IA deduced from the nucleotide sequence was glycine, whereas it was found to be arginine by amino acid sequencing of purified sarcotoxin IA. Analysis of the elution profiles on h.p.l.c. of the synthetic derivatives of sarcotoxin IA showed that the C-terminal amino acid residue of authentic sarcotoxin IA is amidated arginine, which is probably produced by enzymic cleavage of terminal glycine.     

Structure, regulation and function of PKB/AKT--a major therapeutic target

Protein phosphorylation and dephosphorylation play a major role in intracellular signal transduction activated by extracellular stimuli. Protein kinase B (PKB/Akt) is a central player in the signal transduction pathways activated in response to growth factors or insulin and is thought to contribute to several cellular functions including nutrient metabolism, cell growth and apoptosis. Recently, several significant publications have described novel mechanisms used to regulate PKB. Since the alteration of PKB activity is associated with several human diseases, including cancer and diabetes, understanding PKB regulation is an important task if we are to develop successful therapeutics.     

Cloning of non-human primates: the road "less traveled by"

Early studies on cloning of non-human primates by nuclear transfer utilized embryonic blastomeres from preimplantation embryos which resulted in the reproducible birth of live offspring. Soon after, the focus shifted to employing somatic cells as a source of donor nuclei (somatic cell nuclear transfer, SCNT). However, initial efforts were plagued with inefficient nuclear reprogramming and poor embryonic development when standard SCNT methods were utilized. Implementation of several key SCNT modifications was critical to overcome these problems. In particular, a non-invasive method of visualizing the metaphase chromosomes during enucleation was developed to preserve the reprogramming capacity of monkey oocytes. These modifications dramatically improved the efficiency of SCNT, yielding high blastocyst development in vitro. To date, SCNT has been successfully used to derive pluripotent embryonic stem cells (ESCs) from adult monkey skin fibroblasts. These remarkable advances have the potential for development of human autologous ESCs and cures for many human diseases. Reproductive cloning of nonhuman primates by SCNT has not been achieved yet. We have been able to establish several pregnancies with SCNT embryos which, so far, did not progress to term. In this review, we summarize the approaches, obstacles and accomplishments of SCNT in a non-human primate model.     

Rapid generation of knockdown transgenic mice by silencing lentiviral vectors

Lentiviral vectors are potent gene delivery vehicles that enable stable expression of transgenes in both dividing and postmitotic cells, including preimplantation embryos. We have developed lentiviral vectors carrying silencing cassettes consisting of an RNA polymerase III promoter expressing short hairpin RNAs. Transgenic mice can be generated rapidly by transduction of early embryos with lentiviral silencing vectors, resulting in mice with downregulated target genes. We describe two alternative early embryo transduction protocols (removal of zona pellucida and subzonal microinjection). These methodologies offer the possibility of large-scale generation of knockdown transgenic mice for functional genomic studies and enable the production of transgenic mice in 7 weeks.     

Production of Rubratoxin B by Penicillium purpurogenum Stoll

Culture filtrates and chloroform extracts from Penicillium purpurogenum isolated from foodstuffs were subjected to preliminary survey for toxic effects on HeLa cells and mice. The toxic metabolite was isolated in a crystalline form from P. purpurogenum and was identified as rubratoxin B. Pathological findings observed in HeLa cells and mice treated with the metabolite are briefly described.     

Abundance,growth and grazing loss rates of picophytoplankton in Barguzin Bay,Lake Baikal

The abundance, growth, and grazing loss rates of picophytoplankton were investigated in August 2002 in Barguzin Bay, Lake Baikal. Water samples for incubation were taken once at a near-shore station and twice at an offshore station. Contributions of picophytoplankton to total phytoplankton were high (56.9–83.9%) at the offshore station and low (5.8–6.8%) at the near-shore station. The picophytoplankton community in the offshore station comprised mainly phycoerythrin (PE)-rich cyanobacteria, with eukaryotic picophytoplankton being less abundant. In contrast, as well as PE-rich cyanobacteria and eukaryotic picophytoplankton, phycocyanin (PC)-rich cyanobacteria were found in the near-shore station. At the offshore station, growth and grazing loss rates on 25 August were 0.56 and 0.43 day⁻¹, respectively, and on 29 August, 0.69 and 0.83 day⁻¹, respectively. At the near-shore station, growth and grazing loss rates were 1.61 and 0.70 day⁻¹, respectively. These results show that there is a difference in the abundance, composition, and ecological role in the microbial food web of picophytoplankton between the near-shore and the offshore areas in Barguzin Bay.