韩国社会保健事业发展状况研究

该论文主要分析了韩国保健事业的发展现状和韩国人的健康状况。随着近代化的推进，韩国的保健事业有了长足的发展。保健事业的规模不断的扩大，担任保健事业的医疗工作人员的数量日益增多，保健事业的服务水平也逐渐提高，而公共领域的保健事业的发展封改善国民的健康状况发挥了实质性的作用。其中变化最明显的是，作为上个世纪二十年代第一大死亡原因的传染病，因医学技术的发展和保健水平的发展而逐渐排除在十大死亡原因之外。另外，保健部门在自然水普及率的提高、吸烟率的下降等方面卓有成效的工作改善了韩国人健康状态。不过，从平均健康寿命和精神压力等方面看，韩国人的健康状态仍存在不少问题。     

Mechanism of a Decrease in Potency for the Recombinant Influenza A Virus Hemagglutinin H3 Antigen During Storage

The recombinant hemagglutinin (rHA)-based influenza vaccine Flublok® has recently been approved in the United States as an alternative to the traditional egg-derived flu vaccines. Flublok is a purified vaccine with a hemagglutinin content that is threefold higher than standard inactivated influenza vaccines. When rHA derived from an H3N2 influenza virus was expressed, purified, and stored for 1 month, a rapid loss of in vitro potency (~50%) was observed as measured by the single radial immunodiffusion (SRID) assay. A comprehensive characterization of the rHA protein antigen was pursued to identify the potential causes and mechanisms of this potency loss. In addition, the biophysical and chemical stability of the rHA in different formulations and storage conditions was evaluated over time. Results demonstrate that the potency loss over time did not correlate with trends in changes to the higher order structure or hydrodynamic size of the rHA. The most likely mechanism for the early loss of potency was disulfide-mediated cross-linking of rHA, as the formation of non-native disulfide-linked multimers over time correlated well with the observed potency loss. Furthermore, a loss of free thiol content, particularly in specific cysteine residues in the antigen's C-terminus, was correlated with potency loss measured by SRID.     

A Sensitive Method for Quantitation of Rifabutin and Its Desacetyl Metabolite in Human Biological Fluids by High-Performance Liquid Chromatography (HPLC)

Sensitive HPLC-UV methodology has been developed and validated for quantitating rifabutin, an antimycobacterial, and its 25-desacetyl metabolite, LM-565, in human plasma and urine. The HPLC separation for both plasma and urine samples was performed on an ODS, 5-µm, reverse-phase column (25 cm × 4.6-cm ID) using a mobile phase of acetonitrile/0.05 M potassium phosphate, pH 4.2, with triethylamine, (38:61.5:0.5, v/v), at a flow rate of 1.0 ml/min. The separation eluate was monitored by absorbance at 275 nm. Plasma samples (1 ml) were spiked with an internal standard (medazepam), buffered at pH 7.4 and extracted with 80:20 (v/v) hexane:ethyl acetate, and then back extracted with acidified water (0.05 M H3PO4). Linearity was established between 5.0–800 and 2.5–400 ng/ml for rifabutin and LM-565, respectively. Intraday imprecision for rifabutin and LM-565 plasma quality controls prepared at 7.3 and 3.2 ng/ml, respectively, was <15% relative standard deviation (RSD). Absolute recovery for parent drug and metabolite, from plasma, was >90% throughout the respective dynamic ranges and >70% for medazepam. Urine samples (1 ml) were acidified with 50 µl of 3.6 M H2SO4 and diluted with 0.1 M ammonium acetate. Linearity was established between 100 and 5000 ng/ml for both rifabutin and LM-565. Intraday imprecision for a urine control at 200 ng/ml was 12% RSD for either component. The method is currently being used to support Phase I kinetics program for rifabutin in prophylaxis of MAC infection of AIDS patients. Application of this method to a bioavailability assessment is presented.     

Characterization of Esterase and Alcohol Dehydrogenase Activity in Skin. Metabolism of Retinyl Palmitate to Retinol (Vitamin A) During Percutaneous Absorption

Retinyl palmitate, a widely used ingredient in cosmetic products, is promoted for its beneficial effects on the appearance of skin. Previous studies suggest that enzymes are available in skin to metabolize this ingredient during skin absorption. Esterase activity hydrolyzes retinyl palmitate to retinol (vitamin A), which is oxidized in many tissues to retinoic acid primarily by alcohol dehydrogenase. The activities of esterase and alcohol dehydrogenase were characterized in hairless guinea pig skin by using flow-through diffusion cells and radiolabeled model compounds (methyl salicylate and benzyl alcohol) previously shown to be metabolized by these enzymes. Methyl salicylate was hydrolyzed by esterase to a greater extent in viable skin than in nonviable skin. Glycine conjugation of salicylic acid and benzoic acid occurred only in viable skin. The metabolism of methyl salicylate and benzyl alcohol occurred to a greater extent in male guinea pig skin than in female guinea pig skin. The percutaneous absorption of both radiolabeled compounds was similar in viable and nonviable skin. About 30 and 18% of topically applied retinyl palmitate were absorbed from an acetone vehicle by hairless guinea pig skin and human skin, respectively. Less than 1% of the applied dose of this lipophilic compound diffused from skin into the receptor fluid. Retinol was the only detectable metabolite of retinyl palmitate in both hairless guinea pig and human skin. In human skin, 44% of the absorbed retinyl palmitate was hydrolyzed to retinol. The use of retinyl palmitate in cosmetic formulations may result in significant delivery of retinol into the skin.James Boehnlein: Results submitted as partial fulfillment of requirements for the M.S. in Pharmaceutical Science degree (Cosmetic Science),     

Structure-Based Grafting,Mutation, and Optimization of Peptide Inhibitors to Fit in the Active Pocket of Human Secreted Phospholipase A2: Find New Use of Old Peptide Agents with Anti-Inflammatory Activity

Phospholipase A₂ (PLA₂) is a key enzyme in the production of diverse mediators of inflammatory conditions, which possesses an open active pocket that is physicochemically compatible with a variety of small-molecule substrates and peptide inhibitors. Although various peptides and peptide analogues have been identified to have inhibitory activity against PLA₂ originated from animals and plants, only very few were designed for human secreted PLA₂ (hsPLA₂), an attractive target of inflammatory arthritis. Considering that the catalytic domains of PLA₂ family members across different species are highly conserved in primary sequence, advanced structure, and biological function, in this study, we proposed a synthetic pipeline to implement structure-based grafting, mutation, and optimization of peptide ligands from the snake PLA₂–peptide complex crystal structures into the active pocket of apo hsPLA₂ structure to computationally generate a large number of potential peptide inhibitors for hsPLA₂, and the hsPLA₂ inhibitory potency of few highly promising candidates arising from the theoretical analysis was determined. As might be expected, three peptides FLSFK, FLVYK, and FISYR showed relatively high inhibitory capability against hsPLA₂, and other three ALSYK, LVFYA, and KGAILGFM were also modestly potent as they can suppress the enzymatic activity with observable doses. Further, the designed peptide FLVYK with highest potency was carried out with structure-guided modification based on its atomic interactions with hsPLA₂ using the computationally modeled structure data, consequently resulting in a dual-point mutant E L I YK with significantly increased activity.     

Evaluation of Biological Activity of Turkish Plants. Rapid Screening for the Antimicrobial,Antioxidant, and Acetylcholinesterase Inhibitory Potential by TLC Bioautographic Methods

Using thin-layer chromatography (TLC) bioautography, a total of 58 extracts from various organs (aerial parts, leaves, flowers, fruits, roots) of 16 Turkish plants were tested for their antibacterial, antifungal, acetylcholinesterase inhibitory, antioxidant, and radical scavenging activities. The hexane, CHCl₃/CH₂Cl₂, water, and total MeOH extracts were used. No activity was observed against two Gram-negative bacteria (Escherichia coli and Pseudomonas aureginosa) and the yeast Candida albicans. However, 23 plant extracts, mostly the CHCl₃/CH₂Cl₂ and H₂O-solubles, inhibited the growth of all five Gram-positive bacteria tested, Micrococcus luteus, Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Staphylococcus epidermidis. Of the active extracts, the CHCl₃-soluble of the roots of Putoria calabrica (L. fil) DC (Rubiaceae) displayed the highest antibacterial potential. The majority of the CHCl₃/CH₂Cl₂ crude extracts also appeared to inhibit acetylcholinesterase on TLC plates at 100 µg/spot concentration. Particularly active samples were the middle polarity extracts (CHCl₃/CH₂Cl₂) of the leaves of Rhododendron smirnovii Trautv., R. ponticum L., and R. ungernii Trautv. (Ericaceae). β-Carotene, β-carotene/linoleic acid mixture, and 2,2-diphenyl-l-pieryhydrazyl (DPPH) solutions sprayed onto TLC plates were used for detecting antioxidant and radical scavenging properties of the crude extracts. Antioxidant and radical scavenging activities were found to be predominant in highly polar extracts. The water-solubles of all Rhododendron (Ericaceae) and Phlomis (Lamiaceae) species presented the most significant activity.     

A Sensitive and Specific Procedure for Quantitation of ADR-529 in Biological Fluids by High-Performance Liquid Chromatography (HPLC) with Column Switching and Amperometric Detection

An HPLC method using electrochemical detection (ED) has been validated for the determination of ADR-529 in plasma and urine using ICRF-192 as an internal standard (IS). Prior to storage and quantitation, both plasma and urine samples require acid stabilization. Acidified plasma samples were prepared for HPLC using a two column solid-phase extraction (SPE). An aliquot of buffered plasma (i.e., pH 6-7) was first deproteinated and desalted on a C-18 SPE column. The analytes were then eluted onto a C-8 SPE column where retention and selective cleanup were achieved in the cation-exchange mode via silanol interactions. Acidified urine samples were diluted in acetonitrile prior to injection. The HPLC system for plasma and urine samples employed two narrow-bore silica columns used in the weak cation-exchange mode and separated by a switching valve. To prohibit late-eluting peaks from passivating the glassy carbon working electrode, a heart-cut containing ADR-529 and the IS was vented from the first silica column to the second using an automated switching valve. Amperometric detection at an oxidation potential of +1050 mV vs a Ag/AgNO₃ reference electrode was used. Linearity was validated between 5 and 500 µg/ml in plasma and between 2 and 100 µg/ml in urine. Imprecision and percentage bias were typically <10% for both plasma and urine controls throughout their respective dynamic ranges. The absolute recoveries for ADR-529 and the IS from plasma were >95%. This method is being successfully applied to the pharmacokinetic/dynamic evaluation of ADR-529 in animals and humans.     

Biological bases for the integration of appetitive and consummatory grooming behaviors in the cat: a review.

Cats with pontile lesions, frontal neocortical lesions, and thyroidectomized cats display a dissociation of the appetitive and consummatory components of grooming behavior following tactile stimulation of the body surface, an abnormal behavior which waxes and wanes with the seasons of the year. Tryptophan hydroxylase activity and serotonin levels were significantly decreased in the superior colliculi (but not other brain regions) in cats with pontile lesions or frontal neocortical lesions, but not in thyroidectomized cats. Systemic administration of 5-hydroxytryptophan or monoamine oxidase inhibition plus tryptophan administration abolishes the abnormal grooming behavior in each group of cats, and microinjections of 5-hydroxytryptophan or serotonin into the superior colliculi has the same effect, indicating that the change in a serotonergic system is a critical aspect of the abnormal behavior in cats with lesions and that a serotonergic system may also be involved in the genesis of the abnormal grooming behavior in thyroidectomized cats. Functional inactivation of the serotonergic system by p-chlorophenylalanine, LSD, or serotonin receptor blockade does not induce the abnormal grooming behavior in normal cats, indicating that other factors are involved. Cats with lesions and thyroidectomized cats display a rhythmic dysfunction in the excretion of glucocorticoids and glucocorticoid administration abolishes the abnormal grooming behavior, suggesting that glucocorticoids are the other critical factor. Adrenalectomized cats do not display the abnormal grooming behavior, but when adrenalectomized cats are treated with p-chlorophenylalanine, the abnormal behavior appears. Thus, a serotonergic system in the superior colliculi, operating at some level of glucocorticoid function, is involved in the integration of appetitive and consummatory grooming behaviors.     

神经网络用于苯乙胺衍生物的QSAR研究

将神经网络应用于定量构效关系（ＱＳＡＲ）中。采用改进的反向传播算法探讨了肾上腺素能阻断剂Ｎ，Ｎ－二甲基－２－溴苯乙胺取代衍生物的生物活性与取代基疏水参数（Σπ）和电子参数（Σσ）之间的关系。获得了精密的拟合及准确的预测（最大误差小于１０％），优于多无线性回归法。作为一种有效的化学计量学工具（Ｃｈｅｍｏｍｅｔｒｉｃｓ），神经网络具有良好的预测效果及较强的非线性处理功能，可望在ＱＳＡＲ及药物制剂中发挥重要作用。     

微波辅助提取印楝叶多酚及其体外抗氧化活性研究

目的 筛选微波辅助提取印楝叶总多酚的工艺条件,并测定印楝叶总多酚的体外抗氧化活性。方法 通过单因素和响应曲面法设计实验,考察萃取温度、萃取时间、乙醇体积分数3个响应变量以及变量之间交互作用对印楝叶多酚提取效果的影响,对提取工艺进行优化。以清除2,2-联苯基-1-苦基肼基(DPPH·)自由基、羟自由基(·OH)及还原力测定实验评价印楝叶多酚的抗氧化能力。结果 微波辅助印楝叶多酚提取的最佳工艺条件为萃取温度62 ℃,萃取时间40 min,乙醇体积分数48%;响应变量影响顺序为萃取温度>乙醇体积分数>萃取时间;验证实验得到印楝叶中多酚提取得率平均为2.96%,与理论值相对误差为1.33%。印楝叶多酚清除DPPH·自由基的半数抑制质量浓度(IC₅₀)约为2.6 mg·L^-1,·OH自由基的IC₅₀约为15 mg·L^-1,在较低浓度时印楝叶多酚具有较强还原能力。结论 印楝叶多酚在低浓度时表现出较维生素C更强的抗氧化活性。     

高效液相色谱法用于西洋参与白参的分析和鉴别

采用高效液相色谱法测定了１０个不同地区栽培的西洋参与白参中人参皂苷 Ｒｂ１的含量,并利用相对保留时间和相对含量对两者进行定性鉴别     

A Stability-indicating High Performance Liquid Chromatographic Assay for the Simultaneous Determination of Atenolol and Lercanidipine Hydrochloride in Tablets

A simple, rapid, precise and accurate isocratic reversed phase stability indicating HPLC method was developed and validated for the simultaneous determination of atenolol and lercanidipine hydrochloride in commercial tablets. The chromatographic separation was achieved on phenomenex Gemini C18 (250×4.6 mm, 5 μm) column using a mobile phase consisting of acetonitrile and buffer (20 mM potassium dihydrogen phosphate pH 3.5) in the ratio of (55:45, v/v) at a flow rate of 1.0 ml/min and UV detection at 235 nm. The linearity of the proposed method was investigated in the range of 40-160 μg/ml (r(2)=0.9995) for atenolol and 8-32 μg/ml (r(2)=0.9993) for lercanidipine. Degradation products produced as a result of stress studies did not interfere with the detection of atenolol and lercanidipine and the assay can thus be considered stability-indicating.     

Liquid crystalline phases of monoolein and water for topical delivery of cyclosporin A: characterization and study of in vitro and in vivo delivery.

Reverse cubic and hexagonal phases of monoolein have been studied as drug delivery systems. The present study was aimed at investigating whether these systems enhance the cutaneous penetration of cyclosporin A (CysA) in vitro (using porcine ear skin) and in vivo (using hairless mice). Different mesophases were obtained depending on CysA concentration. CysA at 4% allowed the formation of reverse cubic and hexagonal phases in a temperature range of 25-40 degrees C. At 8%, CysA induced the formation of other phases, which might be due to an interaction between the polar groups of the peptide and monoolein. In vitro, the cubic phase increased the penetration of CysA in the stratum corneum (SC) and epidermis plus dermis ([E+D]) at 12 h post-application. The reverse hexagonal phase increased CysA penetration in [E+D] at 6 h and percutaneous delivery at 7.5 h post-application. In vivo, both liquid crystalline phases increased CysA skin penetration. Topical application of these systems, though, induced skin irritation after a 3-day exposure. These results demonstrate that liquid crystalline systems of monoolein are effective in optimizing the delivery of peptides to the skin. The skin irritation observed after topical application of cubic and hexagonal phases should be minimized for their safe use as topical delivery systems.     

In Vitro Diffusion Cell Design and Validation. I. A Stability-Indicating High-Performance Liquid Chromatographic Assay for Betamethasone 17-Valerate in Purified Isopropyl Myristate Receptor Phase

Pharmaceutical Research -     

分散液相微萃取-HPLC测定五味子中五味子甲素和五味子酯甲

目的 采用有机液体分散液相微萃取对五味子药材提取液进行纯化富集,建立分散液相微萃取结合HPLC测定五味子甲素和五味子酯甲的含量。方法 分散液相微萃取以60 μL四氯化碳为萃取剂,供相pH为1,萃取4 min,以转速1500 r.min^-1离心2 min。在上述优化条件下,对五味子药材提取液进行萃取后,以C₁₈柱为固定相、甲醇-0.6%磷酸溶液（69：31）为流动相、检测波长254 nm进行色谱分析。结果 分散液相微萃取对五味子甲素和五味子酯甲的富集倍数分别为5112和1118,线性范围在0.10~19 μg·mL^-1之间,回收率分别为100.1%和94.4%。结论 本法灵敏度高、重现性好,简便、快速、环保,可用于五味子药材中五味子甲素和五味子酯甲的含量测定。     

A Novel Experimental and Theoretical Method for Estimating Albumin-Mediated Hepatic Uptake Based on the Albumin Binding Fraction in Plasma and Human PK Prediction Using a Physiologically-Based Pharmacokinetic Approach

Recently, protein-facilitated uptake has been suggested to be an important factor in the precise prediction of the pharmacokinetic (PK) profiles of drugs. In our previous study, a physiologically-based pharmacokinetic (PBPK) approach considering the mechanism of albumin-mediated hepatic uptake was developed for predicting human PK profiles. It was assumed that drugs affected by albumin-mediated hepatic uptake would bind only to albumin, which means that there would be over-estimation of the contribution of protein-facilitated uptake for a drug that could bind to multiple proteins. In this study, we developed a method that can evaluate the albumin binding fraction in plasma considering the affinity for other proteins. Based on the albumin binding fraction, the contribution of albumin-mediated hepatic uptake was theoretically estimated, and then the human PK profiles were predicted by our proposed PBPK approach incorporating this mechanism. As a result, the predicted human PK profiles agreed well with the observed ones, and the absolute average fold error of PK parameters was almost within a 1.5-fold error on average. These findings show the importance of considering protein-facilitated uptake and also suggest that our proposed PBPK approach can be useful in scientific discussions with regulatory authorities.     

Endocrine-Disrupting Chemicals: Prepubertal Exposures and Effects on Sexual Maturation and Thyroid Activity in the Female Rat. A Focus on the EDSTAC Recommendations

In 1996, the US Environmental Protection Agency was given a mandate by Congress to develop a screening program that would evaluate whether variously identified compounds could affect human health by mimicking or interfering with normal endocrine regulatory functions. Toward this end, the Agency chartered the Endocrine Disruptor Screening and Testing Advisory Committee in October of that year that would serve to recommend a series of in vitro and in vivo protocols designed to provide a comprehensive assessment of a chemical's potential endocrine-disrupting activity. A number of these protocols have undergone subsequent modification by EPA, and this review focuses specifically on the revised in vivo screening procedure recommended under the title Research Protocol for Assessment of Pubertal Development and Thyroid Function in Juvenile Female Rats. Background literature has been provided that summarizes what is currently known about pubertal development in the female rat and the influence of various forms of pharmaceutical and toxicological insult on this process and on thyroid activity. Finally, a section is included that discusses technical issues that should be considered if the specified pubertal endpoints are to be measured and successfully evaluated.