An experiment was conducted to elucidate the effect of graded levels of vitamin $D_3$ in White Leghorn (WL) layer breeders on egg production, shell quality, hatchability of eggs and juvenile performance of offspring during their late laying period (72-88 wk). White Leghorn breeder females were randomly divided into 5 groups of 50 each and were housed in individual California cages in an open-side housing system. Considering birds in five cages as a replicate, 10 such replicates were randomly allotted to each treatment. A basal diet was formulated containing all the nutrients as recommended for WL layers except vitamin $D_3$, which served as control. Another, four diets were formulated by supplementing graded levels of feed grade crystalline cholecalciferol to the basal diet that contained 300, 600, 1,200 and 2,400 ICU of vitamin $D_3$ per kg. Each diet was offered ad libitum to one of the above five treatment groups. The egg production, egg weight, daily feed consumption and the feed intake per dozen eggs or kg egg mass of the birds fed diet without any supplemental vitamin $D_3$ was comparable with those of supplemental groups. Similarly, the level of vitamin in the diet did not have any effect on any of the above parameters. However, the specific gravity of eggs laid by the birds fed the diet without supplemental vitamin $D_3$ was comparable with either 600 or 2,400 ICU supplemental groups but significantly higher when compared to the 300 and 1,200 ICU groups. The egg -shell breaking strength was significantly lowered in the 600 ICU supplemental groups as compared to the strength of other dietary groups. The Haugh unit, egg shell weight, shell thickness, tibia breaking strength, bone ash and calcium content were not influenced by vitamin $D_3$ concentration in the diet. Serum Ca concentration was influenced by vitamin $D_3$ level in the diet. The serum Ca concentration of birds fed either control or the vitamin supplemented diet up to 1200 ICU/kg diet was comparable. However, increasing the concentration of vitamin $D_3$ to 2,400 ICU/kg diet significantly enhanced the concentration of Ca in the serum, which was significantly higher compared to other dietary groups. The serum concentration of P and protein, however, was not influenced by level of vitamin $D_3$ in the diet. Neither fertility nor hatchability was influenced by vitamin $D_3$ concentration in the diet. Feeding a vitamin $D_3$ deficient diet or supplementation of vitamin to hens did not have any influence on their progeny chicks. It can be concluded that dietary supplementation of vitamin $D_3$ may not be essential for optimum production, shell quality, hatchability, and juvenile performance of WL breeders during 72 to 88 weeks of age.
Seven different drying methods were tested in peony roots of Euisung cultivar, harvested in February, in three year's old plant. The roots were selected in length and diameter and half of the samples were removed cork-layers to compare the effects of cork-layer in processing of drying. The 3$0^{\circ}C$ heat drying without cork-layer reduced in days to drying by five days compared to those of the with cork-layers at the same temperature. The 5$0^{\circ}C$ heat drying after cork-layers removed was the most effective in days to drying. In quality of skin color of the 5$0^{\circ}C$ heating was worse to compare with the lower drying temperature. In the drying at room temperature and the heat drying at lower temperature, the paeoniflorin content in drying after cork-layers removed were higher than that of the drying with cork-layers. However, in the boiling water treatment, the paeoniflorin contents in drying after cork-layers removed were lower than those of with cork-layers. In heat drying, paeoniflorin content showed a decreasing tendency to increase of drying temperature. Total sugars in the peony roots showed a decreasing tendency according to the drying temperature increasing, but starch concentration showed a increasing tendency at the same condition. Concentrations of crude protein, crude fiber and crude ash were showed no differences in various drying methods and the materials with or without cork-layers. Relationships between the paeoniflorin and total sugars, and the paeoniflorin and starch were different significantly by the materials of cork-layers removed or not. The 30~4$0^{\circ}C$ heat drying without cork-layers was the most advisable condition for drying in paeoniflorin concentration, days to drying and skin color after drying.
Ultrastructure of gametes in the three-spine stickleback, Gasterosteus aculeatus aculeatus was observed, utilizing light, scanning and transmission electron microscopes. The egg of three-spine stickleback is spherical and demersal type. The eggs are highly adhesived to each other but not to substrates. There are many oil droplets in vitelline membrane. The outer surface of egg envelope is arranged by mushroom-like structures and pore canals. The egg have a micropyle, sperm entry site, in the area of the animal pole. The egg envelope consists of three layers, an outer layer with high electron density, a middle layer consisting two layers and an inner layer consisting of 16 to 20 layers. In the fertilized egg envelope, the molecular weights of these components ranged from 14 kDa to 205 kDa. The molecular weights of nam protein bands are 19.4 kDa, 36.7 KDa, 39.4 kDa, 42.9 kDa, 46.1 kDa and 53.0 kDa. The head of spermatozoa is spherical shape and the acrosome is absent. The mitochondria in midpiece are arranged from one to three layers and separated from the axoneme by the cytoplasmic canal. The tail has two lateral fins and the axoneme is of the 9+2 structure.
MMBR system has been suggested as a promising system to resolve harvesting problems induced from low settling efficiency of microalgae. And recently, a lot of research on reducing fouling at the MMBR system has investigated focused on EPS in many cases. EPS of microalgae mainly consists of polysaccharides and protein components, and is produced through photosynthesis and nitrogen-carbon metabolic pathways. Especially, P-EPS is one of major compounds which occur membrane fouling phenomenon, as its hydrophobic protein components cause floc formation and cake layer accumulation. And it is already known that almost every microalgae can metabolize P-EPS or Chl-a when nitrogen sources as a substrate is insufficient or exhausted situation. With the above backgrounds, uptake rates of P-EPS or Chl-a by Scenedesmus quadricauda according to the type of carbon source and nitrogen concentration were evaluated in order to verify correlation between carbon source vs P-EPS production, and indeed Scenedesmus quadricauda uses P-EPS or Chl-a when the amounts of nitrogen sourc es in the feed is not satisfied. As a result, it was shown that P-EPS and Chl-a production were increased proportional to nitrogen concentration under organic carbon condition. And especially, the amo unts of P-EPS and Chl-a in the cell were diminished with the nitrogen source becomes insufficient or exhausted. Because P-EPS accelerates fouling at the MMBR system, P-EPS degradation by Scenedesmus quadricauda in order to get nitrogen source may contribute to reducing fouling. About a affects of N-consumed Chl-a to the MMBR fouling, more survey is needed. On the contrary, considering the purpose of MMBR system of this study, i.e. harvesting useful high value microalgae efficiently feeding adequate industrial process wastewater, it seems like difficult to maintain satisfied metabolic activity and to harvest with high yield rate using nitrogen-poor MMBR feed.
Yun-Jin Lee;Chang-Hoon Woo;Young-Jun Kim;Hyeon-Ji Kim;Hee-Duk An
Journal of Korean Medicine Rehabilitation
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v.33
no.3
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pp.47-65
/
2023
Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.
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 C1orf31 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.
The herbicide butachlor [N-(butoxymethyl)-2-chloro-N-(2,6-di-methylphenyl) acetamide] is widely used by farmers as a tool for weed management of transplanted rice(Oryza sativa L.) in Taiwan. The herbicide did not stop germination of rice and weed seeds, but strongly inhibited the subsequent growth of young shoots and roots. The inhibition was also strong on established seedlings. However, they could recover to normal growth after the herbicide effect disappeared. Butachlor greatly decreased the endogenous indole-3-acetic acid (IAA) but increased the endogenous abscisic acid (ABA) contents of rice seedlings. Addition of lAA into growth medium (Hoagland's solution) partly relieved growth inhibition. Pretreatment of both gibberellic acid ($GA_3$) and IAA 24 hours before butachlor treatment almost completely alleviated the butachlor-interfere with GA and/or IAA metabolism or their action resulting in the growth inhibition of rice. Butachlor was readily absorbed by rice roots. During 24 hours of uptake experiment, 32% of the applied herbicide was absorbed. Pretreatment of the herbicide for 2 days did ncx affect the absorption. Of the absorbed herbicide, 80% remained in roots, only 20% transported into shoots, and more than 50% was metabolized to water soluble substances. Thin-layer chromatographic (TLC) analysis indicated that the Rf value of the most abundant metabolite was butachlor-glutathione conjugate. Rice, barnyardgrass (Echinochloa crus-galli (L.) Beauv.), and monochoria (Monochoria vaginalis Presl) seedlings contained relatively high level of non-protein thiols, while the glutathione S-transferase (GST) activity was found highest in rice, barnyardgrass the next, monochoria the lowest. The difference in GST activity among these species might be related to their sensitivity to butachlor.
By the activation of ovary hormone, many morphological changes occur in the epithelial cell lines and muscle cells in rat uterus. These two cells in uterus are important to the implantation of embryo, maintaining pregnancy and starting parturition. One important change associated with the morphological change of these two cells in uterus is the change on prostaglandin(PG) metabolism. Its presence and synthesis in endometriurn and myometrium in uterus affects estrous cycle and the start of embryo implantation in uterus. It also performs as an important modulator in parturition. So the abnormally weak expression of PG causes difficulty during labor and over-expression causes pre-term labor. PG biosynthesis starts from either free or liberated arachidonic acids from membrane phospholipid by phospholipase. Such arachidonic acids are converted into PG catalyzed by Cyclooxygenase. Under normal physiological condition, Cyclooxygenase-1(COX-1) having 602 units of amino acids controls the synthesis of PG. It acts as a local hormone regulating vasomodulation of blood flow, flexible muscle movement, increasing the blood permeability and contributing the protective role in preserving integrity of the stomach lining and Cyclooxygenase-2 (COX-2) is induced by the inflammation, pregnancy and increased its expression until parturition. Lipid metabolite like PG is located in uterine and expression of COX-2 increased with pregnancy. Increased expression of COX proteins in epithelial cells and myometrial cells are told to increase the muscle contractility in uterus but decreased right after the labor in rat. It is a good sign indicating that COX proteins are deeply related to the start of labor. Currently, Several studies report the use of PG and COX-2 inhibitor as medication for controlled abortion or to prevent pre-term labor but they entail various side-effects. Our study proposed to suggest use of acupuncture as an another mediator to control abortion or pre-term labor without causing unnecessary side-effects by those medicines. Two acupuncture sites, LI-4 & SP-6 were selected due to their known efficacy. From the immunohistochemical staining of COX-2, normal expression of COX-2 protein in nonpregnant SD rat's uterus revealed that COX-2 protein was primarily detected in the lumina epithelial lining and in the epithelial cell lining contacting the stromal cells. High resolution optical microscopic scanning revealed distinguishable staining in the myometrial mucosa. LI-4 acupuncture administered nonpregnant rat's uterus showed strong expression for COX-2 in endometrium contacted with lumina epithelial lining of rat uterus and in myometrial mucosa. Stromal cells showed more staining than untreated nonpregnant rat's uterus and stronger staining in stromal cells contacting myometrial layer compared to untreated nonpregnant rat's uterus. SP-6 acupuncture administered nonpregnant rat's uterus showed weak expression for COX-2 in myometrial layers and stromal cells but no staining was visible in lumina epitheliai and glandular epithelial cells. Few stromal cells and myometrial mucosa were positively stained for COX-2. Pregnant SD rat's uterus was also immunostained for COX-2 expression after 18 days of pregnancy. Unlike to untreated nonpregnant rat's uterus, luminal epithelial cells were not positively stained for COX-2 but stronger staining for COX-2 was revealed in stromal cells. LI-4 acupunctured SD rat's uterus had very strong expression of COX-2 in luminal epithelial lining. Few stromal cells showed stronger positive COX-2 staining and myometrial layers also showed more expression than untreated pregnant rat. SP-6 acupuncture administered pregnant SD rat's uterus showed positive expression of COX-2 in epithelial cells of luminal mucosa layer but weaker than that of LI-4 acupuncture treatment's case. However, strong positive staining was revealed in stromal mucosa and myometrial layers. Virgin SD rat's uterus motility index during LI-4 acupuncture was 66.52 % (Prob〉T = 0.0197) compared to its motility before the acupuncture treatment but the motility index was slighdy elevated up to 79.58 % (Prob〉T = 0.1175) after the acupuncture. During the SP-6 acupuncture treatment for 30 minutes, uterus motility index was 90.52 % (Prob〉T = 0.1832) showing lesser decrement but consequently reached similar motility index decreasal to 79.95 % (Prob〉T = 0.0215) after the acupuncture treatment as LI-4 showed. LI-4 acupuncture tend to be a quick treatment to reducing the uterus motility in a virgin rat but eventually both two acupuncture administration created very similar reduction of uterus motility seeing the index after the both acupunctures. The uterus movement monitored during the LI-4 acupuncture administered for 30 minutes, Pregnant SD rat showed decreased motility down to 77.90 % (Prob〉 T = 0.0076) compared to uterus motility before the acupuncture and it continuously decreased down to 71.81 %(Prob〉T = 0.0214) after the removal of needle. The statistical analysis using paired t-test showed significance difference for both two motility indexs at =0.05. SP-6 acupuncture administered to pregnant SD rat also had similar pattern of decreasing uterus motility index down to 74.70 % (Prob〉T = 0.1730) during the initial 30 minutes acupuncture administration and it was continuously lowered to 71.52 % (Prob〉T = 0.0155) after the acupuncture. The paired t-test resuit for SP-6 suggest prompt response of uterus motility index to the SP-6 acupuncture treatment but consequently reached same level of inducing the motility reduction as LI-4 at =0.05 level.
Park Hyo-Jung;Park Ha-Joo;Uhm Ki-Nam;Kim Hyung-Kwoun
Microbiology and Biotechnology Letters
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v.34
no.3
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pp.204-210
/
2006
Ethyl (S)-4-chloro-3-hydroxybutyrate is a useful intermediate for the synthesis of Atorvastatin, a chiral drug to hypercholesterolemia. In this research, two 4-chloro-3-hydroxybutyro-nitrile-degrading strains were isolated from soil sample. They were identified as Rhodococcus erythropolis strains by 16S rRNA analysis. The nitrile-degrading enzyme(s) were suggested to be nitrile hydratase and amidase rather than nitrilase from the result of thin layer chromatography analysis. The corresponding genes were obtained by PCR cloning method. The predicted protein sequences had identities more than 96% with nitrile hydratase ${\alpha}-subunit$, nitrile hydratase ${\beta}-subunit$, and amidase of R. erythropolis. The 4-chloro-3-hydroxybutyronitrile-hydrolyzing activities in both strains were increased dramatically by ${\varepsilon}-caprolactam$ which was known as good inducer for nitrile hydratase. Both intact cells and cell-free extract could hydrolyze the nitrile compound. So, the intact cell and the enzymes could be used as potential biocatalyst for the production of 4-chloro-3-hydroxybutyric acid.
Jo, Gyunghee;Jeong, Mun Sik;Wi, Jimin;Kim, Doo Hyun;Kim, Sangkyu;Kim, Dain;Yoon, Jun-Yeol;Chae, Heesu;Kim, Kyun-Hwan;Hong, Hyo Jeong
Journal of Microbiology and Biotechnology
/
v.28
no.8
/
pp.1376-1383
/
2018
The hepatitis B virus (HBV) envelope contains small (S), middle (M), and large (L) proteins. PreS1 of the L protein contains a receptor-binding motif crucial for HBV infection. This motif is highly conserved among 10 HBV genotypes (A-J), making it a potential target for the prevention of HBV infection. In this study, we successfully generated a neutralizing human monoclonal antibody (mAb), 1A8 (IgG1), that recognizes the receptor-binding motif of preS1 using a phage-displayed human synthetic Fab library. Analysis of the antigen-binding activity of 1A8 for different genotypes indicated that it can specifically bind to the preS1 of major HBV genotypes (A-D). Based on Bio-Layer interferometry, the affinity ($K_D$) of 1A8 for the preS1 of genotype C was 3.55 nM. 1A8 immunoprecipitated the hepatitis B virions of genotypes C and D. In an in vitro neutralization assay using HepG2 cells overexpressing the cellular receptor sodium taurocholate cotransporting polypeptide, 1A8 effectively neutralized HBV infection with genotype D. Taken together, the results suggest that 1A8 may neutralize the four HBV genotypes. Considering that genotypes A-D are most prevalent, 1A8 may be a neutralizing human mAb with promising potential in the prevention and treatment of HBV infection.
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