• 제목/요약/키워드: In-vitro techniques

검색결과 347건 처리시간 0.03초

한방방제의 제제학적 연구(1) - 추출방법에 따른 녹용사군자탕의 비교 - (Studies on Pharmaceutical Quality of Oriental Medicinal Preparations (I) - Studies on Decoction of Nokyong-Sagunja-Tang -)

  • 최혁재;김은진;김성수;김남재
    • 생약학회지
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    • 제37권3호
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    • pp.143-150
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    • 2006
  • Decoction of oriental medicinal preparation is prepared in various manners, and changes of chemical constituents might be occurred depending on the processing techniques. The present study was undertaken to investigate the phγsio-chemical and pharmacological equivalence between two extraction methods of Nokyong-sagunja-Tang. Samples were Prepared as follows ; Sample-I was prepared by simultaneously extracting Sagunja-Tang and velvet antler in one vessel. Sample-lI was prepared by adding velvet antler exact to the water extract of Sagunja-Tang. Both sanples showed similar results of physiochemical parameters such as pH, yield, TLC and HPLC chromatogram, and contents of ginsenoside $Rb_1$ and glycyrrhizin. Also, there were little different between two samples in pharmacological effects such as DPPH free radical scavenging effect, and inhibitory effects on xanthine oxidase, hyaluronidase, trypsin, TBA-Rs formation and hemolysis in vitro. And both samples showed no significant difference in antifatigue activities in mice. These results suggest that there might be little difference between two extraction process when velvet antler added to Sagunia-Tang.

한국재래흑염소 수정란의 이식;형질전환 흑염소 생산을 위한 수정란의 채취와 이식 (Embryo transfer in Korean Native Black Goat;Embryo recovery and transfer for the production of transgenic goat)

  • 신상태
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2000년도 춘계심포지움
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    • pp.64-75
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    • 2000
  • During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period (1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor (hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

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Mequitazine의 Muscarine수용체에 대한 작용 (Effect of Mequitazine on the Muscarinic Receptors)

  • 이신웅;장태수
    • Biomolecules & Therapeutics
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    • 제3권3호
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    • pp.192-198
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    • 1995
  • The affinity of mequitazine, a non-sedating antihistamine, for muscarinic receptors was evaluated in the guinea-pig ventricle and ileum by in vitro binding techniques and functional studies. In binding studies, [$^3$H]quinuclidinyl benzilate (QNB) identified a single class of muscarinic receptors with similar apparent $K_{D}$ value of about 100 pM in two tissues. Mequitazine inhibited [$^3$H]QNB binding to muscarinic receptors competitively. Analysis of the mequitazine inhibition curve of [$^3$H]QNB binding to ventricular microsome and ileal homogenate indicated the presence of a single homogeneous binding site with Ki value of 25 nM and 18 nM, respectively. In functional studies, mequitazine caused parallel rightward shifts of concentration-response curves for carbachol and histamine in the isolated guinea-pig ileum. The slope values obtained from Schild plot analysis for the antagonistic action of mequitazine on muscarinic and histamine $H_1$-receptors were not significantly different from unity. The p $A_2$values of mequitazine for muscarinic and histamine $H_1$-receptors were about 7.6 ( $K_{M}$= 25.1 nM) and 8.88 ( $K_{H}$= 1.32 nM), respectively. These results indicate that the muscarinic receptor blocking action of mequitazine is 15 times less potent than the $H_1$receptor blocking action, but high concentration of this drug may cause the peripheral muscarinic receptor blocking effect.t.t.t.

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한국재래흑염소 수정란의 이식: 형질전환 흑염소 생산을 위한 수정란의 채취와 이식 (Embryo transfer in Korean Native Black Goat: Embryo recovery and transfer for the production of transgenic goat)

  • 신상태
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2000년도 수정란의 위생적 처리·검사 및 특수가축의 수정란이식 기술 심포지움
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    • pp.64-75
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    • 2000
  • During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

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Diffusion-based determination of protein homodimerization on reconstituted membrane surfaces

  • Jepson, Tyler A.;Chung, Jean K.
    • BMB Reports
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    • 제54권3호
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    • pp.157-163
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    • 2021
  • The transient interactions between cellular components, particularly on membrane surfaces, are critical in the proper function of many biochemical reactions. For example, many signaling pathways involve dimerization, oligomerization, or other types of clustering of signaling proteins as a key step in the signaling cascade. However, it is often experimentally challenging to directly observe and characterize the molecular mechanisms such interactions-the greatest difficulty lies in the fact that living cells have an unknown number of background processes that may or may not participate in the molecular process of interest, and as a consequence, it is usually impossible to definitively correlate an observation to a well-defined cellular mechanism. One of the experimental methods that can quantitatively capture these interactions is through membrane reconstitution, whereby a lipid bilayer is fabricated to mimic the membrane environment, and the biological components of interest are systematically introduced, without unknown background processes. This configuration allows the extensive use of fluorescence techniques, particularly fluorescence fluctuation spectroscopy and single-molecule fluorescence microscopy. In this review, we describe how the equilibrium diffusion of two proteins, K-Ras4B and the PH domain of Bruton's tyrosine kinase (Btk), on fluid lipid membranes can be used to determine the kinetics of homodimerization reactions.

In Vitro Evaluation of Antioxidant Potential of Date Palm Collected in Algeria using Electrochemical and Spectrophotometrical Techniques

  • Bensaci, Cheyma;Ghiaba, Zineb;Dakmouche, Messaouda;Belfar, Assia;Belguidoum, Mahdi;Bentebba, Fatima Zohra;Saidi, Mokhtar;Hadjadj, Mohamed
    • Korean Chemical Engineering Research
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    • 제59권2호
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    • pp.153-158
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    • 2021
  • In this study, we will determined the total phenolic content (TPC) and the antioxidant activity of the methanolic extract (ME) of date palm (Phoenix dactylifera. L) fruits (DPF) of four native cultivars from Algeria: Ghars (Gh), Chtaya (Cht), DeglaBeïda (DB) and Tinissine (Tns). The TPC of ME of DPF was measured by using Folin-Ciocalteu. Thereafter, the antioxidant capacity of various extracts was determined using DPPH test, reducing power and superoxide anion test. These results showed that dates had strongly scavenging activity on DPPH. The value of IC50 for DPPH radical test was 0.077 mg/ml in Cht. Also, Cht cultivar showed the best-reducing power, which was significantly higher than the other varieties. The less IC50 value in cyclic voltammetry method (CV), which meets the highest effective antioxidant, was 0.006 mg/ml in methanolic extract of Cht.

Enhancement of Efficiency for Polymerase Chain Reaction Using Nanoparticle-Coated Graphene Oxide

  • 주민영;백승훈;김은주;;박찬영;박태정
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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    • pp.375.1-375.1
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    • 2016
  • Polymerase chain reaction (PCR) has revolutionized genetics and become one of the most popular techniques in modern biological and medical sciences. It can be used not only as an in vitro DNA amplification method but also used in many bioassay applications. The PCR can be used to exponentially produce a large number of DNA copies from a small quantity of DNA molecules in a few hours. However, as unwanted DNA fragments are also often manufactured, the amplification efficiency of PCR is decreased. To overcome this limitation, several nanomaterials have been employed to increase the specificity of the PCR reaction. Recently, graphene has attracted a great interest for its excellent electron transfer, thermal and biocompatibility. Especially, gold nanoparticle-coated graphene oxide (GO/AuNPs) led to enhance electron and thermal transfer rate and low-charge transfer resistance. Therefore, we report the development of a demonstration for the PCR efficiency using a large-scale production of the GO and combination of gold nanoparticles. Because a thermal conductivity is an important factor for improving the PCR efficiency in different DNA polymerases and different size samples. When PCR use GO/AuNPs, the result of transmission electron microscopy and real-time quantitative PCR (qPCR) showed an enhanced PCR efficiency. We have demonstrated that GO/AuNPs would be simply outperformed for enhancing the specificity and efficiency of DNA amplification procedure.

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The in vivo significance of in vitro test procedures for the evaluation of drug products

  • Pernarowski, M.
    • 약학회지
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    • 제16권3호
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    • pp.113-120
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    • 1972
  • The last decade of this centry is now the accepted birth date of that sub-discipline of pharmacy that is now called 'biopharmceutics'. Wagner defines biopharmaceutics 'as the study of the influence of fomulation on the therapeutic activity of a drug product.' More specifically, he states that biopharmaceutics encompasses the study of the relationship between the nature and intensity of the biological effects observed in animals or man and the following factors: 1. The nature of the form of the drug (ester, salt, complex, etc). 2. The physical state, particle size, and surface area. 3. Presence or absence of adjuvants with the drug. 4. The type of dosage form in which the drug is administered. 5. The pharmaceutical process (es) used to make the dosage form. The philosophy inherent in this definition has revolutionized our thinking with respect to product development, quality control, and to the practice of pharmacy itself. Althoughthe the emphasis herein will be on quality control, the interrelationship between this and the other areas of pharmacy will be evident. The principles of quality control dictate that a wide variety of techniques be used to evaluate the quality of a dosage form. Since quality must be built into a dosage form, the pharmaceutical scientist begins the process at the research stage, continues it during the production stage, and ends it by applying the tests and procedures established by parmacopeial commissions. These stages are usually separate and distinct and, because of this, product quality has become synonymous with compliance with pharmacopeial specifications.

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Development of Polymeric Blend Microspheres from Chitosan-Hydroxypropylmethyl Cellulose for Controlled Release of an Anti-Cancer Drug

  • Reddy, Lakshmi C. Narayana;Reddy, Rama Subba P.;Rao, Krishna K.S.V.;Subha, M.C.S.;Rao, Chowdoji K.
    • 대한화학회지
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    • 제57권4호
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    • pp.439-446
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    • 2013
  • Chitosan (CS) and hydroxypropylmethyl cellulose (HPMC) blend microspheres were prepared by water-in-oil emulsion technique and were loaded with an anti-cancer drug 5-fluorouracil (5-FU). CS-HPMC microspheres were characterized by Fourier transform infrared spectroscopy to confirm the cross-linking reaction. Scanning electron microscopy (SEM) was also used to assess the surface morphology of particles prepared. The quantity of release of 5-FU from the microspheres have been studied in terms of blend composition and amount of cross-linking agent. Differential scanning calorimetry and X-ray diffraction techniques indicated a uniform distribution of 5-FU particles in microspheres, whereas SEM suggested the spherical structure of the microspheres with slight rough surface. The in vitro drug release indicated that the particle size and release kinetics depend upon blend composition, amount of cross-linking agent used and amount of 5-FU present in the microspheres.

미세조작 및 PCR 기법을 이용한 성판별 수정란의 생산 및 동결 (Production and Cryopreservation of Sexed Embryos after Micromanipulative Biopsy and PCR)

  • 이홍준;서승운;김기동;이상호
    • 한국수정란이식학회지
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    • 제15권2호
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    • pp.175-180
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    • 2000
  • The possible use of micromanipulative biopsy and PCR of the biopsied embryonic cells was tested to produce sexed bovine embryos in practical terms. By micromanipulation and PCR techniques, higher survival rate and accurate sexing of demi-embryos were btained. Bovine oocytes matured and fertilized in vitro were co-cultured with bovine oviductal epithelial cell (BOEC) monolayer in USU-6 medium supplemented with 15% FBS, and the embryos of 37% (327/885) were developed to blastocysts. Among 111 blastocysts produced by invitro, only 7 (6.3%) embryos were found unable to determine their sex, probably due to the loss of cells, since no PCR product was found from those cells. All the remaining 104 (93.7%) demi-embryos survived micromanipulation and demonstrated male-specific product or bovine-specific product alone suggesting that correct sexing of the sample. Forty-three point one percent(25/58) of manipulated and cryopreserved demi-embryos after thawing were survived. Final verification of the sexed embryos is necessary to make sure the same sex in fetus and newborn calf upon embryo transfer. The established sexing method on a large number of bovine embryos from previous and this study suggests that this a could be used practically in the field.

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