In recent years, the interest towards the relationship between incretins and bone has been increasing. Previous studies have suggested that glucagon-like peptide-1 (GLP-1) and its receptor agonists exert beneficial anabolic influence on skeletal metabolism, such as promoting proliferation and differentiation of osteoblasts via entero-osseous-axis. However, little is known regarding the effects of GLP-1 on osteoblast apoptosis and the underlying mechanisms involved. Thus, in the present study, we investigated the effects of liraglutide, a glucagon-like peptide-1 receptor agonist, on apoptosis of murine MC3T3-E1 osteoblastic cells. We confirmed the presence of GLP-1 receptor (GLP-1R) in MC3T3-E1 cells. Our data demonstrated that liraglutide inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as detected by Annexin V/PI and Hoechst 33258 staining and ELISA assays. Moreover, liraglutide upregulated Bcl-2 expression and downregulated Bax expression and caspase-3 activity at intermediate concentration (100 nM) for maximum effect. Further study suggested that liraglutide stimulated the phosphorylation of AKT and enhanced cAMP level, along with decreased phosphorylation of $GSK3{\beta}$, increased ${\beta}-catenin$ phosphorylation at Ser675 site and upregulated nuclear ${\beta}-catenin$ content and transcriptional activity. Pretreatment of cells with the PI3K inhibitor LY294002, PKA inhibitor H89, and siRNAs GLP-1R, ${\beta}-catenin$ abrogated the liraglutide-induced activation of cAMP, AKT, ${\beta}-catenin$, respectively. In conclusion, these findings illustrate that activation of GLP-1 receptor by liraglutide inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation through $cAMP/PKA/{\beta}-catenin$ and $PI3K/Akt/GSK3{\beta}$ signaling pathways.
bone grafting is indicated in the case of bony defects and is classified into autograft, allograft, and xenograft. Synthetic bone graft is contrasted with these three categories in that it has a different donor source. Autograft is most prominent as it is known as a gold standard of all grafting procedures. Its principles and practices are well established via accumulated informations and clinical experiences, which imposes no regulations or restrictions in its clinical use. On the other hand, other bone graft procedures are under tight control for the safety and effectiveness of each product. Food and Drug Administration of the United States has a system in which the information on the approvals and clearances of bone graft materials on their internet homepage. All the bone graft materials that are under the regulations of the United States are classified into the category of medical devices, which includes allogenic bone, xenogenic bone, and synthetic bone graft materials. Each bone graft material has its own indication and the FDA approvals and clearances of medical devices contain the item of "intended use" to specify the indications of each bone graft materials. US dentists, as users of the specific bone graft materials, are provided with adequate information on the approved materials they are to utilize. As an user of these materials, Korean dentists are less provided with the information on the bone graft materials they want to use. Medical providers of the bone graft materials have to be able to provide their users with the essential information such as the intended use of the regulatory approval. Dentists must also be active in gathering informations on the material of their interest, and the system must be built in which both of the medical providers and users of bone graft materials can be satisfied in providing and getting the information, respectively.
In order to detect the way of absorption and metaboism of the urea it is sprayed on the surface of the leaves of sunflower. The sunflowers used in this study are grown in different conditions such that the one in nittogen aboundant and the other in nitrogen deficient soil, respectively. The urea-N, ammonia-N, amide-N, and 80% alcohol soluble-N in the leaves were quantitatively determined. All of the nitrogenous components measured are generally tended to increased with rising the concentration of urea except only amide-N at 24 hours after sprayed, and these were highly significances. It seemed that hydrolizing of urea into ammonia and carbon dixide and the assimilation of ammonia into other organic nitrogenous constituents were rapid in the young leaves than in the mature. It is interest that the amide content, in the young leaves and nitrogen defieient one were enhanced with the increasing concentration of urea, although in the mature leaves it did not show any change in the urea treatment. It is presumed that the assimilation rate of ammonia and the urease activity were lower in the matture leaves than in the young and nitrogen deficient leaves. No significance at 5% level showed all of the nitrogenous components except total nitrogen between nitrogen abundant and deficent leaves. Urea content was a high peak at first 12 hours, ammonia at 48 hours, and amide and alcohol soluble nitrogen at 96 hours, whence decrease4d the content of these constituents gradually. The total nitrogen content is not incrased obviously by only one time of urea spray in this study. When the concentration of urea was relatively high there appeared the wilting spots on t도 edge of leaves. As a whole, it seemed that sprayed urea was rapidly absorbed and taken part in nitrogen metabolism within relatively short period.
Most of the endogenous retroviral genes integrated into the primate genome after the split of New World monkeys in the Oligocene era, approximately 33 million years ago. Because they can change the structure of adjacent genes and move between and within chromosomes they may play important roles in evolutionas well as in many kinds of disease and the creation of genetic polymorphism. Comparative analysis of HERVs (human endogenous retroviruses) and their LTR (long terminal repeat) elements in the primate genomes will help us to understand the possible impact of HERV elements in the evolution and phylogeny of primates. For example, HERV-K LTR and SINE-R elements have been identified that have been subject to recent change in the course of primate evolution. They are specific elements to the human genome and could be related to biological function. The HERV-M element is related to the superfamily of HERV-K and is integrated into the periphilin gene as the truncated form, 5'LTR-gag-pol-3'LTR. PCR and RT-PCR approaches indicated that the insertion of various retrotransposable elements in a common ancestor genome may make different transcript variants in different primate species. Examination of the HERV-W elementrevealed that env fragments were detected on human chromosomes 1, 3-7, 12, 14, 17, 20, and X, whilst the pol fragments were detected on human chromosomes 2-8, 10-15, 20, 21, X, and Y. Bioinformatic blast search showed that almost full-length of the HERV-W family was identified on human chromosomes 1-8, 11-15, 17, 18, 21, and X. Expression analysis of HERV-W genes (gag, pol, and env) in human tissues by RT-PCR indicated that gag and pol were expressed in specific tissues, whilst env was constituitively expressed in all tissues examined. DNA sequence based phylogenetic analysis indicated that the gag, pol and env genes have evolved independently during primate evolution. It will thus be of considerable interest to expand the current HERV gene information of various primates and disease tissues.
Streptomyces coelicolor is a gram-positive soil bacterium which is well known for the production of several antibiotics used in various biotechnological applications. But numerous proteins from its genome are considered hypothetical proteins. Therefore, the present study aimed to reveal the functions of a hypothetical protein from the genome of S. coelicolor. Several bioinformatics tools were employed to predict the structure and function of this protein. Sequence similarity was searched through the available bioinformatics databases to find out the homologous protein. The secondary and tertiary structure were predicted and further validated with quality assessment tools. Furthermore, the active site and the interacting proteins were also explored with the utilization of CASTp and STRING server. The hypothetical protein showed the important biological activity having with two functional domain including POD-like_MBL-fold and rhodanese homology domain. The functional annotation exposed that the selected hypothetical protein could show the hydrolase activity. Furthermore, protein-protein interactions of selected hypothetical protein revealed several functional partners those have the significant role for the bacterial survival. At last, the current study depicts that the annotated hypothetical protein is linked with hydrolase activity which might be of great interest to the further research in bacterial genetics.
Park, Mi-Hyun;Lee, Hye-Ja;Bok, Jeong;Kim, Cheol-Hwan;Hong, Seong-Tshool;Park, Chan;Kimm, Ku-Chan;Oh, Berm-Seok;Lee, Jong-Young
BMB Reports
/
제39권4호
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pp.418-425
/
2006
A human bacterial artificial chromosome (BAC) library was constructed with high molecular weight DNA extracted from the blood of a male Korean. This Korean BAC library contains 100,224 clones of insert size ranging from 70 to 150 kb, with an average size of 86 kb, corresponding to a 2.9-fold redundancy of the genome. The average insert size was determined from 288 randomly selected BAC clones that were well distributed among all the chromosomes. We developed a pooling system and three-step PCR screen for the Korean BAC library to isolate desired BAC clones, and we confirmed its utility using primer pairs designed for one of the clones. The Korean BAC library and screening pools will allow PCR-based screening of the Korean genome for any gene of interest. We also determined the allele types of HLA-DRA and HLA-DRB3 of clone KB55453, located in the HLA class II region on chromosome 6p21.3. The HLA-DRA and DRB3 genes in this clone were identified as the DRA*010202 and DRB3*01010201 types, respectively. The haplotype found in this library will provide useful information in future human disease studies.
Park, Hyo-Jun;Kwon, Chang-Seob;Woo, Joo-Yong;Lee, Gil-Je;Kim, Young-Jin;Paek, Kyung-Hee
The Plant Pathology Journal
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제27권2호
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pp.170-182
/
2011
Plants possess multiple resistance mechanisms that protect themselves against pathogen attack. To identify unknown components of the defense machinery in Arabidopsis, gene-expression changes were monitored in Arabidopsis thaliana under 18 different biotic or abiotic conditions using a DNA microarray representing approximately 25% of all Arabidopsis thaliana genes (www.genevestigator.com). Seventeen genes which are early responsive to salicylic acid (SA) treatment as well as pathogen infection were selected and their T-DNA insertion mutants were obtained from SALK institute. To elucidate the role of each gene in defense response, bacterial pathogen Pseudomonas syringae pv. tomato (Pst) DC3000 was inoculated onto individual T-DNA insertion mutants. Four mutants exhibited decreased resistance and five mutants displayed significantly enhanced resistance against Pst DC3000-infection as measured by change in symptom development as compared to wild-type plants. Among them, member of uridin diphosphate (UDP)-glycosyltransferase (UGT) was of particular interest, since a UGT mutant (At1g05680) showed enhanced resistance to Pst-infection in Arabidopsis. In systemic acquired resistance (SAR) assay, this mutant showed enhanced activation of SAR. Also, the enhanced SAR correlated with increased expression of defense-related gene, AtPR1. These results emphasize that the glycosylation of UGT74E2 is a part of the SA-mediated disease-resistance mechanism.
Nile tilapia (Oreochromis niloticus) are mouth-brooders so that the females holding eggs in their mouth sacrifice their somatic growth for reproduction. For this reason, artificial control of reproduction for the culture of this species has been of interest. Manipulation of photoperiod is an emerging technique for such purpose, but little information is available to establish appropriate photoperiod regime. To obtain necessary basic information, sexually mature females were individually accommodated to glass aquarium, and the spawning activity of these females were monitored for two years under natural photoperiod regime. Female tilapia spawned most frequently on March, April and May when the day length gradually increased from 11 hours to 14 hours and least frequently on September, October, November and December when the day length gradually decreased from 13 hours to less than 10 hours in the first year. The decrease of spawning frequency as day length decreased was also observed in the second year, although the increase of spawning frequency as day length increased was less clear. Spawning of female tilapia was less active when the night was dark due to the disappearance of moonlight (Dark Phase), compared to the Phase of Getting Lighter, Light Phase and Phase of Getting Darker. Results from this study suggest that long day length, particularly increasing phase, is favoured for active spawning of Nile tilapia, and that this species, as a tropical fish species, may utilize changing lunar phases as a secondary environmental cue for reproduction.
Puberty is the developmental period which animals obtain the ability of reproducing sexually for the first time in life. In commercially important aquaculture fish species, the onset of puberty is a matter of major interest due to controlling of sexual maturation to improve broodstock management. To investigate pubertal characteristics of female longtooth grouper (Epinephelus bruneus), specimens were classified into three groups by the bodyweight, including 1, 2, and 3 kg group. Thereafter, we focused on ovarian development and level changes of endocrine regulation factors (GnRH, GTHs, steroid hormone). In the non-breeding season (April), the levels of endocrine regulation factors showed increasing trends in accordance with bodyweight gaining; nevertheless, the oocytes were growth phase belongs to almost peri-nucleous stages in all groups. In the breeding season (June), the levels of endocrine regulation factors were fluctuated that decreases in levels of sbGnRH and $FSH{\beta}$ mRNA expressions along with serum $E_2$ concentrations in 3 kg of group. However, $LH{\beta}$ mRNA expression levels sustained increasing trends by the bodyweight. Moreover, the oocytes developed that 2 kg and 3 kg groups obtained plentiful vitellogenic oocytes while 1 kg group was still composed with greater part of pre-vitellogenic oocytes. Especially, the oocytes of 3 kg group reached over 450 ${\mu}m$ of diameters that indicating possibility to enter the final maturations. These results suggest that the progress of pubertal development in female E. bruneus could be classify into three phases via bodyweight, including pre-puberty (1 kg), early-puberty (2 kg) and puberty (3 kg).
Map-based cloning to find genes of interest, marker-assisted selection (MAS), and marker-assisted breeding (MAB) all require good genetic maps with high reproducible markers. For map construction as well as chromosome assignment, development of single copy PCR-based markers and map integration process are necessary. In this study, the 132 markers (57 STS from BAC-end sequences, 13 STS from RFLP, and 62 SSR) were newly developed as single copy type PCR-based markers. They were used together with 1830 markers previously developed in our lab to construct an integrated map with the Joinmap 3.0 program. This integrated map contained 169 SSR, 354 RFLP, 23 STS from BAC-end sequences, 6 STS from RFLP, 152 AFLP, 51 WRKY, and 99 rRAMP markers on 12 chromosomes. The integrated map contained four genetic maps of two interspecific (Capsicum annuum 'TF68' and C. chinense 'Habanero') and two intraspecific (C. annuum 'CM334' and C. annuum 'Chilsungcho') populations of peppers. This constructed integrated map consisted of 805 markers (map distance of 1858 cM) in interspecific populations and 745 markers (map distance of 1892 cM) in intraspecific populations. The used pepper STS were first developed from end sequences of BAC clones from Capsicum annuum 'CM334'. This integrated map will provide useful information for construction of future pepper genetic maps and for assignment of linkage groups to pepper chromosomes.
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