Inositol 1,4,5-trisphosphate (IP₃) plays an important role in the release of Cα²+ from intracellular stores into the cytoplasm in a variety of cell types. IP₃ translocation dynamics have been studied in response to many types of cell signals. However, the dynamics of cytosolic IP₃ in salivary acinar cells are unclear. A green fluorescent protein (GFP)-tagged pleckstrin homology domain (PHD) was constructed and introduced into a phospholipase C δ1 (PLC δ1) transgenic mouse, and then the salivary acinar cells were isolated. GFP-PHD was heterogeneously localized at the plasma membrane and intracellular organelles in submandibular gland and parotid gland cells. Application of trypsin, a G protein-coupled receptor activator, to the two types of cells caused an increase in GFP fluorescence in the cell cytoplasm. The observed time course of trypsin-evoked IP₃movement in acinar cells was independent of cell polarity, and the fluorescent label showed an immediate increase throughout the cells. These results suggest that GFP-PHD in many tissues of transgenic mice, including non-cultured primary cells, can be used as a model for examination of IP₃intracellular dynamics.

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0～10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500～600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro‐matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM‐199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1 kV/cm for 30 μs in 0.3 M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium‐3 (PZM‐3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM‐3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine‐porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat‐porcine and porcine‐bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

Human embryonic stem (ES) cells retain the capacity for self‐renewal, are pluripotent and differentiate into the three embryonic germ layer cells. The regulatory transcription factors Oct4, Nanog and Sox2 play an important role in maintaining the pluripotency of human ES cells. The aim of this research was to identify unknown genes upregulated in human ES cells along with Oct4, Nanog, and Sox2. This study characterizes an unknown gene, named chromosome 1 open reading frame 31 (C1orf31) mapping to chromosome 1q42.2. The product of C1orf31 is the hypothetical protein LOC388753 having a cytochrome c oxidase subunit VIb (COX6b) motif. In order to compare expression levels of C1orf‐ 31 in human ES cells, human embryoid body cells, vascular angiogenic progenitor cells (VAPCs), cord‐blood endothelial progenitor cells (CB‐EPCs) and somatic cell lines, we performed RT‐PCR analysis. Interestingly, C1orf31 was highly expressed in human ES cells, cancer cell lines and SV40‐immortalized cells. It has a similar expression pattern to the Oct4 gene in human ES cells and cancer cells. Also, the expression level of C1orf31 was shown to be upregulated in the S phase and early G2 phase of synchronized HeLa cells, leading us to purpose that it may be involved in the S/G2 transition process. For these reasons, we assume that C1orf31 may play a role in on differentiation of human ES cells and carcinogenesis.

A full-length sequence of a thrombin inhibitor (designated as hemalin) from the midgut of pathenogenetic Haemaphysalis longicornis has been identified. Sequence analysis shows that this gene belongs to a Kunitz-type family, containing two Kunitz domains with high homology to boophilin, the thrombin inhibitor from Rhipicephalus (Boophilus) microplus. The recombinant protein expressed in insect cells delays bovine plasma clotting time and inhibits both thrombin-induced fibrinogen clotting and platelet aggregation. A 20-kDa protein was detected from the midgut lysate with antiserum against recombinant hemalin. The gene expresses at all stages of the tick except for the egg stage and mainly in the midgut of the female adult tick. Real-time PCR analysis shows that this gene has a distinctly high expression level in the rapid bloodsucking period of the larvae, nymphs, and adults. Disruption of the hemalin gene led to a 2-day extension of the tick blood feeding period, and 27.7% of the ticks did not successfully complete the blood feeding. These findings indicate that the newly indentified thrombin inhibitor from the midgut of H. longicornis might play an important role in tick blood feeding.

Pine wilt is the most important disease of pine trees in Korea, Japan and China. The pathogen causing this disease, the pinewood nematode (Bursaphelenchus xylophylus), is transmitted vectored by adults of some cerambycid beetle species and the Japanese pine sawyer, Monochamus alternatus, is the major vector species in Korea. Although chemical insecticides have been used to kill vector insect and thus prevent transmission of the pathogen, the efficacy is not good. In Japan, to control this insect, an entomopathogenic fungus was studied and developed as an insecticide. This is thought to be the convenient and effective method to control M. alternatus. Recently, there are several reports about the pinewood nematode is vectored by also the pine sawyer, M. saltuarius, in Korea. The objective of this study, therefore, was to isolate and identify entomopathogenic fungi from M. saltuarius cadaver to control it. We collected the cadaver of M. saltuarius and then screened several fungi colonies. The pathogenicity of each fungus was tested using oak longicorn beetle, Moechotypa diphysis, as substitutive insect. M. diphysis is also serious pest to various trees in forest. As the result, only one of them showed high pathogenicity against M. diphysis. Selected fungus was identified by microscopic examination and DNA analysis. Pathogenicity was also evaluated to M. saltuarius.

Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cuneaand its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

To understand geographic genetic variation of the species and relationships among populations of the bumble bee, Bombus ardens, that is utilized as green house pollinator we expanded our investigation by sequencing somewhat longer mitochondrial DNA (mtDNA) fragment, covering some uninvestigated regions within the species distribution, and analyzing the sequence data in terms of population genetic structure. For the purpose of study, a portion of mitochondrial COI gene, corresponding to "DNA Barcode" region (658 bp) was sequenced from 160 individuals of B. ardens collected over 15 localities in Korea. The sequence data revealed overall relatively low genetic diversity within species, with a maximum sequence divergence of 0.3%. Geographically, one haplotype (BARBA01) was found in all localities surveyed, with the frequency of 91% (145 among 160 individuals), whereas other haplotypes were found in a locality mostly as a single individual, suggesting that haplotype distribution can be summarized as coexistence of widespread, one dominant haplotype and regionally restricted, other haplotypes. Overall, very high rate of per generation female migration (Nm = 4.6 ~ infinite) and very low level of geographic substitution (FST = 0 ~ 0.099) among localities were characteristic. Although some populations were genetically subdivided from the remaining localities in the hierarchical analysis, there was regional polarity on this subdivision. Taken together with gene flow estimates, the nature of genetic divergence of the bumble bee populations is characterized as one that possessing low genetic diversity, high gene flow, and wide spread of one dominant haplotype, consistent with the previous finding. To have further detailed information of this valuable genetic resource, further longer and variable molecular portion is under investigating.

A recent study on Neuroptera brought us an attention to a newly found group, Coniopterygidae, dustywings. As we reported for the first time this year, this family has not been taxonomically reported in South Korea while it has been reported in North Korea before. In fact, this is known to be found in Japan and China, which means this probably have been around us for a long time. However, we found there was one species of which the name was once mentioned in a paper in 1978. It was reported by Kim, H.S. et al. in 1978 in a study of citrus red mite and its natural enemies (Kim et al., 1978). Although the spelling of the species was wrong even as a synonym, the species was found to be a natural enemy of citrus red mite, Panonychus citri in Jeju-do. We here report this taxonomically undescribed species for the first time in South Korea. The species is superficially similar to white flies but, unlike white flies, it is on our side as a natural enemy.

Lycorma delicatula, once mistakenly reported its occurrence in Korea, is now suddenly common in western Korea, due to its recent arrival from China and their subsequent settlement. A history of name changes in two fulgorid species, Lycorma delicatula and Limois emelianovi is reviewed. We propose to use 꽃매미 instead of its temporary name, 주홍날개꽃매미 for Lycorma delicatula, and, based on the ICZN code 32.5.1, to use Limois emelianovi instead of Limois emeljanovi for 희조꽃매미.

The absolute age of lapilli tuff in sedimentary formation that contains dinosaur fossils in the Boseong area, Korea was determined radiometrically against volcanic rocks below and above the fossil-bearing horizons. The sanidine in the lapilli tuff below the fossil-bearing horizon (Seonso formation) has an 40Ar-39Ar age of 81.l±1.4Ma. The Pilbong tuff above Seonso formation has an 40Ar-39Ar age of 81.0±2.4Ma. An andesite dyke intruding all sedimentary units yields an 40Ar-39Ar age of 42.4±2.5Ma. Thus 81 Ma age can be regarded as the best estimate for the age of the Seonso Formation and the associated the dinosaur eggs. This age correlates well with dinosaur fossil finds in the Haenam and Koseong regions of Korea. The occurrence of dinosaur eggs and clutches attests to the existence of dinosaurs in southern Korea at least inCampanian times.