In general, the mean silkmoth lifespan is around 8 days for female and 5 days for male. But, the duration of J037 strain's lifespan is remarkably long in both sexes. On the contrary, the Daizo(sdi) strain has a remarkably short lifetime. The differences in adult lifetime among various silkworm strains has been suggested that the adult lifetime may be genetically controlled. In this experiment, using J037 and Daizo strains we investigated genetic factors related to the adult lifetime of silkworm. We constructed the full-length cDNA library from the adult male of the J037 strain. A total of 2,688 clones were randomly selected, and we performed a differential display hybridization with cDNA probes generated from J037 and Daizo adult males. In conclusion, 193 clones were identified as differential expressed genes, and 154 unique genes were generated after the assembly of 193 clones. Of the 154 unique genes, the most abundant genes were cytochrome oxidase subunit-1 gene(9 times) and unknown(clone ID; 1-50) gene(5 times). The functional groups of these unique genes with matches in the AmiGo database were constructed according to their putative molecular functions. Among thirteen functional categories, the largest group was unclassified protein(24%). In addition, we analyzed the nucleotide and deduced amino acid sequences of the most highly occurred gene(1-50, EF434397), which consisted of 240 amino acids. However, it is confirmed yet that these genes really have an affected on the silkworms longevity. Further studies on these molecules biological roles will give us well-fined information about mechanisms of insect aging and/or scenesence.
It has been wel known that phospholipase C(PLC) plays an important role in the intracellular signaling in a variety of cell types. However, involvement of PLC in mouse oocyte maturation and preimplantation embryo development remains unknown. The present study examined the expression patterns of the mouse PLC \beta 1 and \gamma 1 during oocyte maturatio and preimplantation embryo development study examined the expression patterns of the mouse PLC \beta 1 and \gamma 1 during oocyte maturation and preimplantation embryo development by the competitive reverse transcription-polymerase chain reaction (RT-PCR method). PLC \gamma 1 mRNA (0.1 fg) was readily detected in germinal vesicle (GV)-stage oocyte and its level was reduced as meiotic resumption proceeded. PLC-\beta 1 mRNA (<0.1 fg) as detected at low level at GV-stage oocytes and scarcely detected at germinal vescle breakdown (GVBD)-stage oocytes. After fertilization, both PLC \beta 1 and \gamma 1 mRNA levels began to increase at morula-stage embryos (0.2 fg) and were more prominent in blastocyst-stage embryos(1 fg). to elucidate the possible involvement of PLC via protein kinase C(PKC) pathway during oocyte maturation and preimplantation embryo development , the effects of sphingosine (PKC inhibitor), sn-$diC_{8}$(PKC activator) anc U73122 (PLC ingibitor) were examined. Treatment of GV-stage oocytes with sphingosine (20 \mu M) facilitated the meiotic resuption by 10-20 over the control within 1 h as judged by GVBD, whereas U73122 failed to show any significant effect. U73122 (10 \mu M) effectively blocked the compaction of morula, while sn-$diC_{8}$(50 \mu M). In summary, the present study shows that the mouse PLC \beta 1 and \gamma 1 are expressed in a developmental stage-specific manner and PLC-PKC pathway may be involved in early preimplantation embryo development.
Park, Jun Mo;Yoo, Hye Jung;Kim, Han Chul;Han, Geul Soon
The Korean Journal of Nuclear Medicine Technology
/
v.16
no.2
/
pp.139-148
/
2012
Purpose : Since standard solution is the one that knows its exact concentration, the curve of the dissolution has been determined according to the amount of the solution, compared to the amount of the unknown sample. Therefore, the antigen that makes up standard materials should be made in a pure form. The configuration of the standard substance solution in the kit we use is a freeze-dried material, or made and comes as a liquid. Lyophilized reference material is used after dissolving in usually D.W. (Distilled Water), and if the antigen to use is too sensitive, reagents should be freeze-dried. Furthermore, when freeze-dried reference has to be frozen again after being dissolved, it should be kept under $-20^{\circ}C$ until the expiration date according to the reports. Since it is not expressed in the experiment if it is safe or stable to reuse the solution which was dissolved a few times, thus, this time it is tested and evaluated that the changes of the standard solution by freezing and melting several times, and its results and the effectiveness of it were compared to the solution which was kept in a fridge. Materials and Methods : Among Vitro diagnostic kits on the market made by radioimmunoassay, parathyroid hormone (PTH), adrenocorticotropic hormone (ACTH), luteinizing hormone (LH) are made of freeze-dried standard solution and all composed of the same Lot.NO. These hormones melted in D.W. and were separated into three groups. In the first group, melting and freezing were repeated, and in the second group, The solution only for one time use was put into a test tube after melting and freeze it. The third group was kept in the refrigerator. This experiment has been conducted from January to February in 2012. January to 2012. PH test was employed because ph is prone to changing depending on the change of protein. Each group of the standard solution, cpm (counter per minute), and the patient relative concentration values were compared by date, and Through the correlation coefficient and Paired t-test, the significant level of each group was analyzed. Results : ACTH, PTH, LH pH values were too subtle denaturation rather than numerical changes in the protein. In addition, when the standard solution of ACTH, PTH, LH was refrigerated, after 3 days and 7 days, there was a significant difference observed between the solution being kept in a refrigerator and a freezer within a significance level. Conclusion : Standard solution should be kept in a freezer, and being kept in a fridge, it is recommended to use the solution as soon as possible.
A 371 agricultural households from 26 different communities in South Korea was subjected on a study of food taboos in January of 1966. To the pregnant women, those to whom a high protein diet is particurally important, as many as 14 different kinds of foods, mostly portein rich foods, were avoided to eat. It is believed that if duck is eaten while pregnant her baby may walk like a duck in later life. Some mother have a strong aversion to the rabbit meat that her unborn baby must be a harelip. It is feared to eat chicken, shark or carp by the pregnant mother for her baby may get a gooseflesh appearance, or fish scale-like skin in later life. It is thought that if mother eats soup made of meat borns, especially chicken bones, a disfigured baby may be born. Some area informed that if mother eats crab meat her future baby will always bubble. To the child-bearing mothers 13 different kinds of foods were avoided to eat. Some believe that if raddish kimchi, soybean curd, squash are eaten while dilivery that mother may get dental decay or to lose all her teeth. Other think that highly spiced raddish kimchi cause delivery difficult. To the lactating mothers 7 different items of foods were not recommended to eat. It is a common belief that eating green vegetables, especially fresh lettuce, are restricted that her baby may stool greenish. It is said that eating ginsen-chicken soup, or ginsen tea during lactating reduces breast milk secretion. To the weaning babies 7 different kinds of foods were prohibited to fee. Eggs are not eaten because mothers think her babies will start to talk very late. Eight different items of foods in cases of gastro-intestinal diseases, 5 items for liver disease, 7 items for high blood pressure as well as for paralysis were respectively restricted. It is said that meats including pork, beef, and chicken are neither desirable for the patients of high blood pressure nor those of paralysis. To the measles children 10 varieties of foods were restricted. Especially soybean products and meats were not encouraged to use for avoiding asecond attack of measles. For the common cold 8 different kinds of foods were aversed and men think that eating of soup of undria delays a recovery. For the tuberculosis 4 kinds of foods were prohibited to eat. It is said that wine, red pepper and ginsen will stimulate lung bleeding. Many mothers had a strong aversion to fermented shrimp and fish in case of style. and 5 different items of foods were restricted. In case of menstration not so many foods were restricted as other cases, but meat soup is not eaten in this condition in some areas. Majority of food taboos in Korean villages are neither based on tribal nor religious factors. But no one knows how, since what ages, from where, these food taboos have been transmitted and spread over the country. This survey found a great variety of food taboos, aversions, traditional beliefs and prohibitions latent unknown reseasons, or non-scientific conceptions, or completely different ideas from the modern medical aspect, or somewhat fallacious and superstitious beliefs. For the vascular disease contrasting approach were found between modern the oritical therapy and popular remedy among the rural populations who largely depend on the eastern medication. Further scientific study on either side should be done to lead the patient proper way. Many restricted foods such as rabbit, duck, chicken and fish are best resources of protein rich foods which are available in the village. Emphasis should be laid upon breaking down fallacious and supersititious food taboos through the extended nutrition education activities in order to improve food habit and good eating pattern for healthier and stronger generations of Korea.
The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.
Kim, Yong Hyun;Yoon, Hyoung Kyu;Kim, Chi Hong;Ahn, Joong Hyun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Park, Sung Hak;Song, Jeong Sup;Cho, Kyung Sook
Tuberculosis and Respiratory Diseases
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v.58
no.6
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pp.590-599
/
2005
Object : Cigarette smoking is a major cause of mucus hypersecretion, which is a pathophysiological feature of many inflammatory airway diseases. Mucins, which are an important part of the airway mucus, are synthesized from the Muc gene in airway epithelial cells. However, the signaling pathways for cigarette smoke-induced mucin synthesis are unknown. The aim of this study was to determine the signal pathway for smoking induced Muc5ac gene expression. Methods : A549 cells were cultured and transiently transfected with the Muc5ac promoter fragment. These cells were stimulated with 5% cigarette smoke extract (CSE) alone or with CSE after a pretreatment with various signal transduction pathway inhibitors (AG1478, PD98059 and SB203580). The Muc5ac promoter activity was examined using the luciferase reporter system, and the level of phosphorylated EGFR, ERK1/2, p38 MAPK and JNK were all examined using Western blot analysis. Muc5ac mRNA expression was also examined using reverse transcriptase polymerase chain reactions (RT-PCR). Results : 1. The peak level of luciferase activity of the Muc5ac promoter was observed at 5% concentration and after 3 hours of incubation with the CSE. The level of EGFR phosphorylation and the luciferase activity of the transfected cells caused by the CSE were significantly suppressed by AG1478 or PD98059 (P<0.01). 2. CSE phosphorylated ERK1/2 or p38 MAPK but not JNK. The Muc5ac mRNA expression level was increased by the CSE but that was suppressed by PD98059 or AG1478. 3. The CSE-induced phosphorylation of ERK1/2 was blocked by PD98059 and that of p38 MAPK was blocked by either PD98059 or SB203580. Either PD98059 or SB203580 suppressed the luciferase activity of the transfected cells (P<0.0001). Conclusion : The Muc5ac mRNA expression level was increased by the CSE. The increased CSE-induced transcriptional activity was mediated via EGF receptor activation, which led to ERK1/2 and p38 MAPK phosphorylation.
Mutations in Fused in Sarcoma (FUS) have been identified in patients with amyotrophic lateral sclerosis (ALS) or Frontotemporal Dementia (FTD). Pathological FUS is mis-localized to cytosol and forms aggregates associated with stress granules (SG), while FUS is normally localized to nucleus. However, it is largely unknown how pathological FUS forms SG-aggregates and which domains are responsible for this process. In this study, we examined cellular localization and aggregation of ALS-linked FUS missense mutants (P525L, R521C, R521H, R521G), analyzed the domains responsible for cytosolic FUS aggregation in HEK293T cells, and confirmed this in cultured mouse neurons. To do this, we firstly generated missense mutants of FUS and then examined their cellular localization. We found that P525L was mostly mis-localized to cytosol and formed FUS-positive SG aggregates while R521C, R521H, or R521G was localized to both nucleus and cytosol. To further characterize the domains required for aggregate formation of cytosolic FUS, we generated different domain-deletion mutants using FUS-∆17 which has a deletion of nuclear localization signal. Interestingly, cytosolic FUS without SYGQ and RGG1 domain or cytosolic FUS without RGG2-ZnF-RGG3 domain did not form FUS-positive SG aggregates, while cytosolic FUS without RRM domain generated more aggregates compared to FUS-∆17. Taken together, these data suggest that SYGQ-RGG1 or RGG2-ZnF-RGG3 domain contributes to formation of cytosolic aggregate, while RRM domain might interfere with FUS aggregation. Therefore, our studies will provide important insight for understanding cellular pathogenesis of neurodegeneration associated with FUS aggregate as well as finding therapeutic targets for ALS or FTD.
Rainbow trout were reared in a stainless steel aquarium from Nov. 11, 1977 to June 12, 1978, and the following results were obtained : 1. The volume of water was about $400\iota$ in a aquarium measuring $1m\;(Length)\times1m\;(Width)\times67cm(Height)$ and water depth 40 cm. Water was supplied for about 16 hours daily at a rate $3\iota/min$ and was drained through the conical settling part in the middle of the aquarium bottom. Filter tank was about $23cm(W)\times23cm(L)\times40cm(D)$ and contained pebbles 30 cm in depth. Water recirculation rate was at)out $1,030\iota/hr$, or 2.6 turn-over per hour. 2. During the first period (77 days), the trout grew from 88.3g to 229g in average, the total weight attaining 30.7kg. The food coefficient was 1.249, average daily increment 243.3g, average daily growth rate 1.245%, and the mortality was 2 smallest fish weighing 53 g, owing to unknown reason. During the second period (135 days), the trout grew from 239g to 555g in average, the total weight attaining 57.2 kg. The food coefficient was 1.447, average daily increment 279.8g, average daily growth rate $0.65\%$ and the mortality was 31 fish weighing 11,255 g, owing partly to miss-handling and partly to disease. 3. The feed consisting of fully domestic materials was prepared in this laboratory, and the feed conversion was not inferior to high protein commercial feed available in foreign countries. 4. The result of whole period for 212 days was 56.5 kg in gross increment, and based on this result, when $1\iota/min$ full day inflowing water available, the net production will become 28.25 kg. So, if a 5000kg production is planned, $180\iota/min$ or about $10.8m^3/hr$ be reauired, and the production in value frill become 15million won at local price at the expense of about 5.3 million won. From the result of this experiment, rainbow trout is feasible for commercial production in Korea with relatively small amount of well water and simplified water recirculation system.
Vitellogenin(Vg) is a sex specific serum protein present in sexually maturing female blood of oviparous vertebrates. Estrogen($E_2$) is a main inducer of hepatic Vg synthesis. We investigated the effects of androgen and growth hormone(GH) on regulation of Vg and estrogen receptor(ER) genes in Japanese eel. Immature eels($200{\sim}250\;g$) were given a single injection of $E_2(5{\sim}5,000\;{\mu}g/kg\;bw)$ alone, or in combination with eel recombinant GH(eGH, $1{\sim}10\;{\mu}g/kg$) or methyltestosterone(MT, $1{\sim}5\;mg/kg$) and sacrificed 10 days after the hormone treatments. Expression levels of ER and Vg genes from the liver were determined by means of reverse transcription and polymerase chain reaction(RT-PCR). Administration of $E_2$ stimulated Vg gene expression in a dose dependent manner. Levels of Vg mRNA after the injection of $E_2(500\;{\mu}g/kg)$ with MT(5mg/kg) or eGH($10\;{\mu}g/kg$) were much higher than in that of $E_2$ alone($500\;{\mu}g/kg$). Whereas, injection of either vehicle, eGH ($10\;{\mu}g/kg$) or MT(5mg/kg) alone did not induce the expression of Vg gene in the liver. ER mRNA was detected from the fish treated with vehicle alone. $E_2$ injection($5{\sim}500\;{\mu}g/kg\;bw$) increased this ER expression but dose dependent response was not clear. Addition of MT(5mg/kg) or eGH($10\;{\mu}g/kg$) did not affect $E_2-stimulated$ ER mRNA expression. This study confirms the necessity of $E_2$ as the primary factor for Vg gene expression and requirement of additional hormones such as MT or GH for the full expression of Vg mRNA, and suggests that the additive effect of MT or GH on Vg gene expression would be mediated by some unknown factors other than ER.
Tak, Byung-Yun;Chung, Myung-Sook;Joo, Kyoung-Hwan;Rim, Han-Jong
Journal of agricultural medicine and community health
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v.16
no.2
/
pp.134-140
/
1991
The amino acid constituents of Paragonimus westermani were very imperfectively known. Lee(1964) detected 9 amino acids in the tissue hydrolysates of P. westermani, and 13 amino acids were detected from the cyst content and body fluid constituents of P. ohirai, But, the quantity of amino acids in P. westermani is still unknown. In the present investigation 18 amino acids, the fundamental constituents of proteins, were quantitatively studied by high performance liquid chromatography. The results obtained were as follows : A total of 18 amino acids were recovered in protein hydrolysates of P. westermani obtained from cat. ; They were cystein, aspartic acid, glutamic acid, serine. glycine, histidine. arginine, threonine, alanine, proline, tyrosine, valine, methionine, isoleucine, leucine, phenylalanine, tryptophan and lysin. Among them, glutamic acid was the most abundant form and tryptophan, cystein, methionine, and histidine constitute minor portion of hydrolysates. Compared to the normal P. westermani, the volume of hydrolysates obtained from the praziquantel(PZQ) treated worm-0.lug PZQ/ml saline for 6 hours incubation, and $3{\times}25$mg/kg bwt${\times}$2days in vivo treatment was generally increased except tryptophan. A total of 17 free amino acids were identified and the volume was 174.18 umol/1 gm wet weight P. westermaini. Among them, glycine and alanine constitute 28% of total volume. No significant differences were observed in the material obtained from worm treated with PZQ. However, slight increase of serine, arginine and the slight diminution of glutamic acid and proline was observed.
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