• The Korean Journal of Zoology
• /
• v.16 no.1
• /
• pp.25-41
• /
• 1973

Ultrastructural changes in the cells of the corpora allata of the pine moth, Dendrolimus spectabilis Butler, were studied by electron microscope to know the structural correlates of hormone production by the gland during the larval-pupal-adult transformations. Mitochondria are in active phases from the overwintered to the last instar larvae and from the pupae just after pupation to the 20-day old pupae, while they are in inactive phases from the making cocoon stage to the prepupae just before pupation. The peripheral allatum cells have electron dense granules in the intracellular vacuoles of smooth-surfaced endoplasmic reticulum in the larval life, particularly in the overwintered larvae and in the early adults but the swollen smooth-surfaced intracytoplasmic vacuoles made by expansion of an end of the tubular rough endoplasmic reticulum, some of which contain fibrous proteins, are observed in addition to the vacuoles in the intercellular spaces in which the vacuoles grow by fusing each other from the mature larvae to the prepupae, both of them disappearing during just before pupation. After pupation the cytolasmic vacuoles develop again in the allatum cells so that they seem to begin the secretory activity. The fact that the neurosecretory granules stored within the axons terminated in the corpus allatum are visible only from the 20-day old pupa about two days before abult emergence to the 5-day old adult means that the secretion from the allatum cells is under the control of the brain from the late pupal stage, while the secretion during from the larval to the early pupal life has no relation with the brain, because such granules are not observed within the axons. It is, therefore, suggested that at least two kinds of hormone are released with the ages as far as concerned with the production and secretion mechanisms of the allatum hormone: juvenile hormone is released until the last instar larvae without any direct stimlation of the brain and gonadotropic hormone is secreted from the late pupa to the adult by getting brain's stimulation and that the secretory phases observed from the mature larvae to prepupae are presumably concerned with the biosynthesis of protein owing to the ecdysone and those from the early pupal stage in uncontrolled condition of the brain with the prothoracotropic activity.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
• /
• 2001.06a
• /
• pp.1031-1031
• /
• 2001

The mammary gland is made up of remarkably sensitive tissue, which has the capability of producing a large volume of secretion, milk, under normal or healthy conditions. When bacteria enter the gland and establish an infection (mastitis), inflammation is initiated accompanied by an influx of white cells from the blood stream, by altered secretory function, and changes in the volume and composition of secretion. Cell numbers in milk are closely associated with inflammation and udder health. These somatic cell counts (SCC) are accepted as the international standard measurement of milk quality in dairy and for mastitis diagnosis. NIR Spectra of unhomogenized composite milk samples from 14 cows (healthy and mastitic), 7days after parturition and during the next 30 days of lactation were measured. Different multivariate analysis techniques were used to diagnose the disease at very early stage and determine how the spectral properties of milk vary with its composition and animal health. PLS model for prediction of somatic cell count (SCC) based on NIR milk spectra was made. The best accuracy of determination for the 1100-2500nm range was found using smoothed absorbance data and 10 PLS factors. The standard error of prediction for independent validation set of samples was 0.382, correlation coefficient 0.854 and the variation coefficient 7.63%. It has been found that SCC determination by NIR milk spectra was indirect and based on the related changes in milk composition. From the spectral changes, we learned that when mastitis occurred, the most significant factors that simultaneously influenced milk spectra were alteration of milk proteins and changes in ionic concentration of milk. It was consistent with the results we obtained further when applied 2DCOS. Two-dimensional correlation analysis of NIR milk spectra was done to assess the changes in milk composition, which occur when somatic cell count (SCC) levels vary. The synchronous correlation map revealed that when SCC increases, protein levels increase while water and lactose levels decrease. Results from the analysis of the asynchronous plot indicated that changes in water and fat absorptions occur before other milk components. In addition, the technique was used to assess the changes in milk during a period when SCC levels do not vary appreciably. Results indicated that milk components are in equilibrium and no appreciable change in a given component was seen with respect to another. This was found in both healthy and mastitic animals. However, milk components were found to vary with SCC content regardless of the range considered. This important finding demonstrates that 2-D correlation analysis may be used to track even subtle changes in milk composition in individual cows. To find out the right threshold for SCC when used for mastitis diagnosis at cow level, classification of milk samples was performed using soft independent modeling of class analogy (SIMCA) and different spectral data pretreatment. Two levels of SCC - 200 000 cells/$m\ell$ and 300 000 cells/$m\ell$, respectively, were set up and compared as thresholds to discriminate between healthy and mastitic cows. The best detection accuracy was found with 200 000 cells/$m\ell$ as threshold for mastitis and smoothed absorbance data: - 98% of the milk samples in the calibration set and 87% of the samples in the independent test set were correctly classified. When the spectral information was studied it was found that the successful mastitis diagnosis was based on reviling the spectral changes related to the corresponding changes in milk composition. NIRS combined with different ways of spectral data ruining can provide faster and nondestructive alternative to current methods for mastitis diagnosis and a new inside into disease understanding at molecular level.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.6
• /
• pp.878-885
• /
• 2017

Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.45 no.2
• /
• pp.181-187
• /
• 2016

Algae is a potential resource with various biological activities. In this study, the anti-inflammatory effect of Grateloupia imbricata Holmes ethanol extract (GIHEE) from red algae was investigated in LPS-induced RAW 264.7 cells. As a result, reduced secretion of pro-inflammatory cytokines [tumor necrosis factors-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6] and nitric oxide (NO) was observed in a dose-dependent manner. Expression of nuclear factor-kappaB (NF-${\kappa}B$) as well as inducible NO synthase and cyclooxygenase-2 proteins was reduced by GIHEE, suggesting that the anti-inflammatory activity of GIHEE is related to suppression of NF-${\kappa}B$ signaling pathways. In addition, GIHEE reduced phosphorylation of mitogen-activated protein kinases. These results suggest that GIHEE can be used as a potential anti-inflammatory therapeutic.

• Journal of Embryo Transfer
• /
• v.17 no.3
• /
• pp.171-177
• /
• 2002

Glucocorticoid is activating mammary gland cells for lactating animals, resulting in increasing abilities of the milk synthesis. Expression of the prolactin receptor(PRL-R) in mammary gland cells was closely associated with milk production. To increase lactation ability for the Korean Native Goats at mid-lactation period. 0.05, 0.1. and 0.2 g of hydrocortisone was administrated with 5 $m\ell$ of saline. and injected into vein. For the control, 5 $m\ell$ of saline was administrated in to vein. After 24 H, the mammary gland tissue was collected, and mRNA expression rates were investigated for the alpha-casein and PRL-R using competitive PCR(polymerase chain reaction). There was no significant differences between treatment and control groups for the mRNA expression rate of PRL-R in mammary gland cells after 24 h of administration of hydrocortisone. The rate of mRNA expression for the alpha-casein was increased 37%, 630%, and 380% at 0.05, 0.1, and 0.2 g of hydrocortisone administration groups, respectively, comparing with control group. The results suggested that PR L-R mRNA expression of mammary gland cell by administration of hydrocortison was not significant, but increase of the alpha-casein mRNA expression my be differences of expression of functional proteins in the cell and expression patterns of protein secretion time to out of the cell. This study showed increase of alpha-casein mRNA expression by administration of hydrocortisone at mid-lactation period of Korean native goat.

• Parasites, Hosts and Diseases
• /
• v.56 no.5
• /
• pp.409-418
• /
• 2018

Acanthamoeba spp. are free-living protozoa that are opportunistic pathogens for humans. Cysteine proteases of Acanthamoeba have been partially characterized, but their biochemical and functional properties are not clearly understood yet. In this study, we isolated a gene encoding cysteine protease of A. castellanii (AcCP) and its biochemical and functional properties were analyzed. Sequence analysis of AcCP suggests that this enzyme is a typical cathepsin L family cysteine protease, which shares similar structural characteristics with other cathepsin L-like enzymes. The recombinant AcCP showed enzymatic activity in acidic conditions with an optimum at pH 4.0. The recombinant enzyme effectively hydrolyzed human proteins including hemoglobin, albumin, immunoglobuins A and G, and fibronectin at acidic pH. AcCP mainly localized in lysosomal compartment and its expression was observed in both trophozoites and cysts. AcCP was also identified in cultured medium of A. castellanii. Considering to lysosomal localization, secretion or release by trophozoites and continuous expression in trophozoites and cysts, the enzyme could be a multifunctional enzyme that plays important biological functions for nutrition, development and pathogenicity of A. castellanii. These results also imply that AcCP can be a promising target for development of chemotherapeutic drug for Acanthamoeba infections.

• Microbiology and Biotechnology Letters
• /
• v.26 no.5
• /
• pp.406-412
• /
• 1998

The endoglucanase gene, egl6, of Trichoderma sp. was connected with the yeast ADH1 promoter, and the resultant plasmid, pVT-C4, was introduced into three S. cerevisiae host strains (YNN27, 2805, and SEY2102). Among each 80 transformants, the cell growth and expression level of endoglucanase were compared in test-tube cultivation, and three respective transformants for each host cells showing the highest expression level and cell growth were selected. When three recombinant yeast cells were batchwise cultivated for 48 hr in flask, the total activities of endoglucanase expressed were about 1140 unit/l with 2805/pVT-C4, 1020 unit/l with SEY2102/pVT-C4, and 590 unit/l with YNN27/pVT-C4. Irrespective of host strain, about 80% of the expressed endoglucanase was detected in the extracellular medium. In addition, it was also found that the recombinant enzyme was secreted into the culture medium as two major forms of lightly and heavily glycosylated proteins.

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.47-53
• /
• 2010

Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

• Journal of Life Science
• /
• v.19 no.3
• /
• pp.343-348
• /
• 2009

Magnaporthe oryzae, a major cause of rice blast, is one of the most destructive plant fungal pathogens. Secretion of reactive oxygen species (ROS) during the infection phase of plant pathogenic fungus plays a key role in the defense mechanism of a plant. ROS causes oxidative damage and functional modification to the proteins in a pathogenic fungus. Methionine, especially, is a major target of ROS, which oxidizes it to methionine sulfoxide. To survive from the attack of ROS, plant pathogenic fungus has antioxidative systems - one example would be methionine sulfoxide reductase B (MSRB), which reverses the oxidative alteration of methionine to methionine sulfoxide. In the present study, identification and molecular characterization of the MSRB gene in M. oryzae KJ201 were investigated. The MSRB gene was amplified by PCR from the M. oryzae KJ201 genomic DNA. The copy number of MSRB in the genome of M. oryzae KJ201 was identified by Southern blot analysis, which revealed that the gene exists as a single copy. To study the molecular function of an MSRB gene, the expression level of the MSRB gene was assayed with hydrogen peroxide treatment by Northern blot analysis and RT-PCR. The expression of the MSRB gene was increased by treatment of hydrogen peroxide, without significant correlation to hydrogen peroxide concentrations. These results indicate that the MSRB gene in M. oryzae KJ201 could contribute to protection against plant defense compounds such as ROS and offer a novel strategy for the control of rice blast.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.1
• /
• pp.135-141
• /
• 1997

The early studies demonstrated that the relative amount of FSH was important for stimulating normal ovarian activity and demonstrated the existence of a threshold level for FSH, above which follicular growth was activated. It was found that only a modest increase in circulating FSH level above the threshold (between 10 and 30%) was required to stimulate folliculogenesis. In addition, FSH is primary responsible for initiating estradiol production through the activation of the aromatase enzyme system in granulosa cells, follicular secretion and growth. LH on the other hand, plays a supportive role in ovarian steroidogenesis, stimulating the ovarian thecal cells to produce androgen, the precursor for estradiol synthesis. But there is now an increasing number of reports in the literature demonstrating an adverse effect of LH on fertility and miscarriage in infertile and fertile women. So HP-FSH is the drug of a highly purified FSH preparation which has a higher specific activity and far fewer impurities than FSH. This study was performed to evaluate the efficacy and safety of HP-FSH administered (SC; subcutaneous) versus FSH(IM; intramuscular) for ovulation induction. 20 candidates patients for ovulation induction were participated. All patients underwent pituitary desensitizing with a long gonadotropin-releasing hormone (GnRH) agonist protocol and ovulation induction was started with HP-FSH SC (10 patients; group I) or FSH IM (10 patients; group II). After ovulation, outcome of ovulation induction and local reaction of injection site were compared. There were no difference of outcome of ovulation in two groups except pregnancy rate/embryo transfer. Group I had a higher pregnancy rate/ embryo transfer than Group II (44.4% Vs 28.6%). Pain, redness, tenderness, bruising and itching when the injection received on the first 5 days of treated (50 SC and 50 IM injections) were assessed. There were no significant difference (P>0.05) in the incidence of tenderness, bruising and itching between the IM and SC injection. But IM injection (FSH) had a tendency of higher above incidence. The number of reports of pain, redness were significantly increased in IM injection group (P<0.05). These results indicate that SC administration of HP-FSH has been shown to be as effect for superovulation as traditional gonadotropins, with an improved safety profile due to the removal of extaneous proteins.