Cysteinylglycinase was partially purified from Proteus mirabilis by consecutive procedure. The specific activity was increased about 16-fold to that of cell-free extract. The enzyme was found rather unstable on ammonium sulfate precipitation ann the precipitated enzyme protein became partially insoluble during dialysis. The precipitated enzyme was found to be solubilized by treatment of 4% Triton X-100 effectiviely, The optimum temperature and pH of the enzyme activity were 35$^{\circ}C$ and 7.3, respectively. After heat treatment of the enzyme at 5$0^{\circ}C$ for 30 min, it lost the activity to 70%. The enzyme was stable at pH 7.0-8.0. The molecular weight of the cysteinylglycinase was found to be about 190,000 by Sephadex G-150 gel filtration. The enzyme was activated by the addition of Mn$^{2+}$ and $Mg^{2+}$ ions. The maximal activation was obtained in preincubation with $Mg^{2+}$ ion for 30 min. The enzyme catalyzed the hydrolysis of various dipeptides and tripeptides. The Km and Vmax values for cysteinylglycine were 1.60 mM and 0.24 m unit/ mg, respectively.
In order to achieve high-level expression of interferon-${\alpha}1$ (IFN-${\alpha}1$) during fed-batch fermentation of recombinant E. coli, effects of oxygen supply and induction temperature on the expression of recombinant proteins were evaluated. Supplementation of oxygen and its transfer into cells is one of the most important parameters involved in the design and operation of mixing-sparging equipment for bioreactors. Generally, higher oxygen supply stimulates cell growth of aerobic microorganism and consequently the amount of products is increased. In this study, the optimum aeration strategy for the higher production of IFN-${\alpha}1$ during fed-batch fermentation of recombinant E. coli was surveyed. The growth of the cells was also monitored with four different concentrations of dissolved oxygen (DO; limiting, 20%, 35%, 50%) conditions. The DO was controlled by varying aeration rates of air and pure oxygen. Oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR) were evaluated and compared for the enhanced growth and induction of the cells and IFN-${\alpha}1$, respectively. We confirmed that increased DO by additional oxygen supply, up to 35%, can improve the production of IFN-${\alpha}1$ during the fermentation.
Kim, Jee-Yeon;Kim, Sung-Eun;Kim, Jae-Eun;Lee, Jong-Chan;Yoon, Je-Yong
Journal of Korean Society of Environmental Engineers
/
v.27
no.7
/
pp.771-776
/
2005
In recent days, there is much interest in the biocidal activity of silver since silver is known to be safe and effective as disinfectant and biocidal material against coliforms and viruses. In particular, nano silted silver particles which can be used as effective biocidal material received more attention. Accordingly, it is important to investigate antimicrobial activity and mechanism of nano sized silver particles prepared in a cost-effective manner. In this study, nano sized silver particles were prepared via photoreduction of a silver salt ($AgNO_3$) in the bulk phase of $PEO_{20}-PPO_{70}-PEO_{20}$ (Pluronic 123) block copolymer The antimicrobial efficacy of silver nano particles against E. coli was investigated and compared with that of silver ion as the concentration of silver nano particles, pH ($5.6{\sim}8.2$), temperature ($4^{\circ}C{\sim}35^{\circ}C$) varied in aqueous system. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) was used to examine the nature of damaged microorganism with nano sized silver particles and silver ion. This study showed that antimicrobial efficacy of silver nano particles was approximately one twentieth than that of silver ion. It was more biocidal at higher pH in contrast with silver ion. In addition, nano silver particles was demonstrated to disrupt the outer membrane of E. coli, subsequently causing their aggregation. On the other hand, silver ion diffused into the cell damaging the cytoplasmic membrane without disrupting the outer membrane of E. coli.
Ciliates have the possibility of a new live food in marine finfish culture because of their wide range of body size, thin tell wall, show motility, and fast reproduction rate. In this research, six species of ciliates were isolated from south coast and salt pond in Korea. The fitness of these species as a live food was evaluated in terms of size, motility, suspensibility and cell density. As the result, Euplotes sp. (K-1) was found suitable to be a new live food which might substitute rotifers, Brachionus plintilis and B. rotundiformis in fish larvae culture. The modified $F{\emptyset}yn's$ Erdschreiberd media, MErds-2 with the addition of glycine, glucose and yeast extract increased six times higher growth rate of Euplotes sp. (K-1) than the basic F$\emptyset$yn's Erdschreiberd media. The optimum water temperature, pH and light intensity for this ciliates were $22.5^{\circ}C$, 8 and 2,000 lux, respectively, and its culture environmental range was relatively wide, On the other hand, this ciliate fed baker's yeast, Saccharomyces cererisiae grew up to 1,240 inds./mL with the inocula of 100 inds./mL within 7 days. The results of the study showed that Euplotes sp. (K-1) has a potential to be utilized as a new live food in fish larvae culture.
Journal of the Society of Cosmetic Scientists of Korea
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v.44
no.1
/
pp.73-79
/
2018
The primary cilium protrudes from the cell body like a bio-antenna that has many receptors, channels and signaling molecules to sense and response to external stimuli. The external environment such as ultraviolet irradiation, temperature, humidity, gravity and shear stress always influences skin. Skin responds to external stimuli and differentiates by making melanin, collagen and horny layer. Ciliogenesis participates in developmental processes of skin, such as keratinocyte differentiation and hair formation. And it was reported that skin pigmentation was inhibited when ciliogenesis was induced by sonic hedgehog-smoothened-GLI2 signaling. When skin is exposed to ultraviolet irradiation, alpha-melanocyte stimulating hormones (${\alpha}$-MSH) increase melanin synthesis through activation of the cAMP pathway in melanocytes. We observed that ${\alpha}$-MSH and cAMP production inducers inhibited ciliogenesis of melanocytes. Therefore, we thought that regulation of ciliogenesis is potential candidate target for the development of agents to treat undesirable hyperpigmentation of skin. As a result, we found out that an ethanol extract of Glycyrrhiza glabra (EGG) root and 3,4,5-trimethoxy cinnamate thymol ester (TCTE, Melasolv) significantly inhibit melanin synthesis of normal human melanocyte by inducing primary cilium formation. This study proposed new theory to regulate skin pigmentation and cosmetic components for skin whitening.
the basic model of chemical reactor using glow discharge, we used cathode discharge cell with vacant cavity in the middle. Currently glow discharge is widely studied as a radiation source or atomization device in atomic spectroscopy and remarkable technological achievements are made through the graft with other analysis devices such as microanalysis and steel analysis.1 Additionally, as the characteristics of basic glow discharge and radiation have been reviewed many times, those results could be used in this experiment.2-3 In 1993, an article regarding the treatment of poisonous gas in the air using low temperature plasma was published. According to this article, if DC Glow Discharge is used under continuous atmospheric flow, poisonous gases such as SO2 and NO can be removed.4 Based on those findings, we designed highly efficient reactor where stable air plasma is composed and all air flow pass the negative glow area passing through the tube. It was observed that the cathode tube type glow discharge developed in this study would be economical, easy to use and could be used as radiation source as well.
The influence of phenolsulfunate concentrations on electroplating characteristics and electrochemical behaviors was investigated with a viewpoint of electrolyte aging using the circulation cell and potentiostate And comparison of tinplate coating appearance such as glossiness and Image clarify has been also studied with varying of phenolsulfonic acid (PSA) solutions. As the aging of electrolyte proceeded, the limiting current density was moved to a lower current density region by the limitation of mass transfer, and higher phenolsulfunate concentrations resulted in the narrower optimum current density range and deterioration of coating surface of tinplates. The difference of the limiting current density was not remarkable with increasing electrolyte temperature. Thus the electrolyte aging was attributed to the limitation of thermally-activated process such as mass transfer of reducible ions. It has also been considered that the accumulation of phenolsulfonate suppressed normal electrotinning reaction by reducing the mobility of stannous ions, taking into account of the smaller effect of electrolyte aging. Experiments showed similar polarization behavior between the electrolyte of high phenolsufonate solution and the aged one, which comes to conclude that the accumulation of phenolsulfonate is one of the major causes of electrolyte aging.
Kim, Sang-Hee;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
International Journal of Oral Biology
/
v.31
no.4
/
pp.141-148
/
2006
The effects of adenosine triphosphate(ATP) on salivary glands have been recognized since 1982. The presence of purinergic recepetors(P2Rs) that mediate the effects of ATP in various tissues, including parotid and submandibular salivary gland, has been supported by the cloning of receptor cDNAs and the expression of the receptor proteins. P2Rs have many subtypes, and the activation of these receptor subtypes increase intracellular $Ca^{2+}$, a key ion in the regulation of the secretion in the salivary gland. The apical pores of taste buds in circumvallate and foliate papillae are surrounded by the saliva from von Ebner salivary gland(vEG). Thus, it is important how the secretion of vEG is controlled. This study was designed to elucidate the roles of P2Rs on salivary secretion of vEG. Male Sprague-Dawley rats (about 200 g) were used for this experiment. vEG-rich tissues were obtained from dissecting $500-1,000\;{\mu}m$ thick posterior tongue slices under stereomicroscope view. P2Rs mRNA in vEG acinar cells were identified with RT-PCR. To observe the change in intracellular $Ca^{2+}$ activity, we employed $Ca^{2+}-ion$ specific fluorescence analysis with fura-2. Single acinar cells and cell clusters were isolated by a sequential trypsin/collagenase treatment and were loaded with $10\;{\mu}M$ fura -2 AM for 60 minutes at room temperature. Several agonists and antagonists were used to test a receptor specificity. RT-PCR revealed that the mRNAs of $P2X_4$, $P2Y_1$, $P2Y_2$ and $P2Y_3$ are expressed in vEG acinar cells. The intracellular calcium activity was increased in response to $10\;{\mu}M$ ATP, a P2Rs agonist, and 2-MeSATP, a $P2Y_1$ and $P2Y_2R$ agonist. However, $300\;{\mu}M\;{\alpha}{\beta}-MeATP$, a $P2X_1$ and $P2X_3R$ agonist, did not elicit the response. The responses elicited by $10\;{\mu}M$ ATP and UTP, a $P2Y_2R$ agonists, were maintained when extracellular calcium was removed. $10\;{\mu}M$ suramin, a P2XR antagonist, and reactive blue 2, a P2YR antagonist, partially blocked ATP-induced response. However, when extracellular calciums were removed, suramin did not abolish the responses elicited by ATP. These results suggest that P2Rs play an important role in salivary secretion of vEG acinar cells and the effects of ATP on vEG salivary secretion may be mediated by $P2X_4$, $P2Y_1$, $P2Y_2$, and/or $P2Y_3$.
Background: Pneumonia is commonly seen in outpatient clinics. it is widely known as the most common cause of death from infectious disease. Pneumonia has been diagnosed by its typical symptoms, chest X-ray and blood tests. However, both chest X-rays and blood tests have limitations in diagnosis. Thus primary care clinicians usually have been constrained due to a lack of adequate diagnostic tools. Vibration response imaging (VRI) is a newly emerging diagnostic modality, and its procedure is non-invasive, radiation-free, and easy to handle. This study was designed to evaluate the diagnostic usefulness of the VRI test among pneumonia patients and to consider its correlation with other conventional tests such as Chest X-ray, laboratory tests and clinical symptoms. Methods: VRI was performed in 46 patients diagnosed with pneumonia in Konkuk University Medical Center. VRI was assessed in a private and quiet room twice: before and after the treatment. Sensors for VRI were placed on a patient's back at regular intervals; they detected pulmonary vibration energy produced when respiration occurred and presented as specific images. Any modifications either in chest X-ray, C-reactive protein (CRP), white blood cell count (WBC) or body temperature were compared with changes in VRI image during a given time course. Results: VRI, chest X-ray and CRP scores were significantly improved after treatment. Correlation between VRI and other tests was not clearly indicated among all patients. But relatively severe pneumonia patients showed correlations between VRI and chest X-ray, as well as between VRI and CRP. Conclusion: This study demonstrates that VRI can be safely applied to patients with pneumonia.
Human being can't live without nature, then the changes of nature affect human body. It means that human body has corresponding changes to the KI(vital energy) of nature. There is a stream of changes in human body which circulate mysteriously and punctually by the laws of nature. If this stream of changes fits into human's life style, it would be most effective. It has a certain mode continuously. So if a person has a habit fitting into it, he will get the healthiest body. Then the researcher tries to explain the changes in human body by the time, mainly focused on within 24 hours. it is showing not only the oriental view, but also the western's. The researcher can find the coincidence as followings. At In-Si(3-5 am), the body function and the body temperature get to the bottom, therefore it's good for him to wake up and to run the vital energy. At Sa-Si(9-11 am), the patience on pain anxiety and the psychic concentration get to the top, he'd better start the work. At O-Si(11am-1pm), the heart energy has a vital move, then the blood concentration of Hb(hemoglobin) gets to the top. At Mi-Si(1-3 pm), the muscle strength, the squeeze, and the breathing rate increase. The reflex nerve sensitivity gets to the top. Creativity, observation, and working efficiency go high, so it's time to work hard. At Hae-Si(9pm-1am), the body function falls, sleeping is needed. At Chuck-Si(1-3 am), the cell spontaneity gets to the top, immune lymphocyte moves actively, and the blood concentration of growth hormone gets to the top. These are liver's work. In west, there has been active studies on how to reduce the side effect by using a person's bio-rhythm based on the 'time treatment', and how to reorganize the bio-rhythm by using the machine and the age resistance based on the 'bio-watch'. Though the 'time treatment' means something, the artificial resistance on bio-rhythm seems to give bad effects to human body. If a person lives by regimen of oriental medicine, he will maintain the healthiest body. Regimen is that human body follows the laws of nature, and moves its mysterious, Punctual and periodical changes.
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