• Archives of Pharmacal Research
• /
• 제19권6호
• /
• pp.486-489
• /
• 1996

A simple, rapid, specific and sensitive method for the detection of serum hepatitis C virus (HCV) RNA using the reverse transcription-polymerase chain reaction (RT-PCR) technique without conventional RNA extraction was developed. HCV template RNA from serum was obtained by boiling the serum at $95^{\circ}C$ for 2 min, cooling rapidly in ice and removing the proteins by cetrifugation. RT-PCR amplifications including the reverse transcription and first PCR amplification were performed in one vessel containing both of reverse transcriptase and Taq DNA polymerase. The detection of HCV RNA from $10^{-3}{\mu}l$. serum was possible with this method. The suitability of this method for clinical analysis was evaluated by assaying HCV RNA in 225 patient samples including anti-HCV antibody negatives (13 samples) and positives (212 samples) by enzyme-linked immunosorbent assay test (ELISA). Detections of HCV RNA with this method were in 4 of 13 anti-HCV antibody negative samples (30.8%) and 95 of 212 positive samples (44.8%). The present method can be completed in 1 hr and has a wide range of application for the clinical utilities to determine the viral RNAS.

• Archives of Pharmacal Research
• /
• 제19권6호
• /
• pp.469-474
• /
• 1996

A rapid and sensitive method has been developed to detect hepatitis B virus DNA (HBV) by nested polymerase chain reaction (PCR) technique with primers specific for the surface and core regions in capillary thermal cycler within 80 min. The lower limit for detection by present PCR method is $10^{-5}$ pg of recombinant HBV DNA which is equivalent to that determined by one round of PCR amplification and Southern blot hybridization analysis. When boiled HBV positive serum was serially diluted 10-fold, HBV DNA was successfully determined in $1{\mu}l-10^{-3}$ of serum. HBV DNA was detected by present method in 69 clinical samples including HBsAg positives and negatives by enzyme-linked immunosorbent assay (ELISA). When serum samples were amplified by nested PCR using surface and core region primers, HBV DNAs were detected in 37 of 69 samples (53.6%) and 18 of 69 samples (26.1%), respectively. These results can inform the infectious state of HBsAg positive pateints. A simple and rapid nested PCR protocol by using boiled serum as DNA template has been described for the clinical utility to determine HBV DNA in human serum.

• Journal of Nutrition and Health
• /
• 제17권1호
• /
• pp.31-40
• /
• 1984

혐기적 조건밑에서 $FeCl_{3}$에 의해 촉진된 TBA 반응을 이용하여 혈청중 과산화물 측정을 위한 새로운미량 분석법을 설정하였다. 즉. 혈청단백침전물이 현탁된 TBA반응흔합액에 혈청 $10{\mu}l$당 $1.0{\mu}mole$의 $FeCl_{3}$를 첨가한 다음 질소기류밑에서 50분간 끊는 물에 중탕한다. 이 분석법의 반응 예민도는 현행 일반 TBA법보다 40여배나 증대되었으며 따라서 미량의 시료$(10{\sim}20{\mu}l)$로도 TBA반응생성물의 농도 측정이 비색법으로도 가능하였다. 그리고 TBA반응생성물의 butanol 추출액을 $(NH_{4})_{2}SO_{4}$염석으로 부분말수시킴으로써 예민성 및 재현성이 개선될 수 있었다. 이 분석법을 이용하여 측정한 건강인들의 혈청 TBA같은 높은 연령군일수록 증가하는 경향을 보였다.

• 대한본초학회지
• /
• 제24권4호
• /
• pp.159-164
• /
• 2009

Objectives : The purpose of this study is to evaluate antioxidant activities and reducing serum alcohol concentration of extract of Lentinus edodes on the alcohol administered rats. Methods : Antioxidant effect was measured by total phenolic compound and DPPH-radical scavenging activity of extract of Lentinus edodes in vitro. Blood alcohol concentration, aldehyde concentration, malondialdehyde concentration, glutathion concentration were measured in vivo. Results : The extract of Lentinus edodes increased DPPH-radical scavenging activity dose-dependently. The water extract with boiling water showed lower antioxidant activity and phenolic content than 70% ethanol extract in vitro. Blood alcohol concentration was significantly reduced by pre-treatment of ethanol extract of Lentinus edodes. The effect was more significant than commercial product used as a positive control. Conclusions : This study suggest that Lentinus edodes can be a potential nature resource for the management of ethanol toxicity although the mechanism of action involved in the treatment remains to be explored.

• 미생물학회지
• /
• 제21권1호
• /
• pp.15-26
• /
• 1983

Commercial deerhorns were extracted in boiling water, the deerhorn extracts were per os introduced into rabbits, and then the effects of the extracts on the antibody productions aginst Escherichia coli antigens were investigated for 4 weeks. The experimental rabbits were divided into 4 groups ; control, only deerhorn, only antigen, and antigen plus deerhorn treated groups. The effects of the treatments were measured by counting the number of blood cells, weighing, and immunoelectrophoresis. The rabbits' body weights gained up to 185% in the deerhorn group, and the other groups gained nearly 120%. The numbers of red blood cells in the antigen plus deerhorn group increased somewhat. However, the numbers of leucocytes gradually increased after one week in the antigen group, and at 4 weeks increased up to 290%. In the antigen plus deerhorn group the numbers of leucocytes increased suddenly up to 189% at one week, but after one week the numbers recovered to normal state. Strangely in the deerhorn group the numbers decreased up to 40%. The amounts of serum globulin increased in the control after one week, but maintained about 130%. In the deerhorn group the amounts increased like the control, but after 4 weeks increased up to 175%. In the antigen group the amounts were not changed until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum .gamma.-globulin decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum ${\gamma}-globulin$ decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 3 weeks, but after 4 week the amounts slightly increased up to 110%. However, the amounts in the deerhorn plus antigen group did not change until 2 weeks, but after 2 weeks abruptly increased up to 174%. The recognized immunoglobulins were IgG and IgM, and the enhanced immunoglobulin was IgG.

• 한국식품영양학회지
• /
• 제7권2호
• /
• pp.108-113
• /
• 1994

This study was carried out to evaluate the effect of Cnidi rhizoma (CR) water extract on fat accumulation In fatted rats induced by the oral high fat administration for six weeks. To accomplish this evaluation, the serum and liver tissue have been examined for enzyme activity, cortisol and insulin level. The change of liver or tissue have been observed by the light microscope. GOT GPT and LDH activities were lower than the control group. Insulin and cortisol were higher than the control group, due to the fat accumulation. The liver of the control group observed by the tight microscope appeared to the fatty liver, but CR group showed some improvement of the fatty liver Based on the above results, it was shown that it is possible to improve fat accumulation induced by high fat dietary through using the oral administration of Cnidi rhizoma water extract.

• Natural Product Sciences
• /
• 제19권4호
• /
• pp.303-310
• /
• 2013

Velvet deer antler (VDA) is well known oriental medicine claimed to have tonic activities as improving bone mineral density (BMD), immune-enhancing, rejuvenating and many other medicinal activities. Ossified deer antler (ODA) is bony product produced by over-calcification of deer antler due to late harvesting. The extraction efficiency of ODA by conventional boiling in water must be very poor due to bony nature, hence the reputations for the medicinal efficacies of ODA has been highly under-evaluated compared to that of VDA without any experimental evidences. Employing our new efficient water extraction process ($135^{\circ}C$), the extracts of ODA and VDA were analysed to compare the contents of bioactive components and the potencies of pharmacological activities. The results showed that; 1) The $135^{\circ}C$ extraction (autoclaving) of ODA gave highly increased amount of biomass, 120% more than the conventional extraction by 100-boiling, whereas the same treatment for VDA showed only 15% increased amount of biomass. 2) Feeding the ODA- or VDA-extracts to oophorectomized rats showed very potent BMD-recovering activity. 3) During the ossification of deer antler, the total collagen content was found to be increased by addition of type-1 to pre-existing type-2 collagen, but not replacement of type-2 to type-1 collagen. High titer of peptide hormones like growth hormone and IGF-1 were detected in the ODA- and VDA-extracts and also in the serum of ODA- or VDA-treated oophorectomized animals dose-dependently. Present experimental data will give a conclusion that folkloric poor reputations on ODA must be concerned only with poor extraction efficiency of conventional $100^{\circ}C$ water extraction and not based on the composition of bioactive substances of ODA.

• 한국식품과학회지
• /
• 제32권4호
• /
• pp.959-963
• /
• 2000

콩은 우유, 계란과 더불어 우리나라에서 과민성 알레르기를 일으키는 대표적인 식품중의 하나로 손꼽힌다. 이에 본 연구에서는 콩에 알레르기 반응을 일으키는 환아의 혈청과 콩 단백질을 토끼에 면역하여 얻은 다클론 항체를 이용하여 immunoblotting 및 ELISA 방법을 통하여 가열 처리한 콩과의 반응성을 조사 함으로서 가열처리가 콩의 항원성 내지는 알레르기성에 미치는 영향을 조사하였다. 물중탕으로 1 시간까지 가열처리 후 SDS-PAGE에 의해 분리된 콩 단백질은 열처리에 따른 콩 단백질 band의 변화를 관찰할 수 없었다. 또한 다클론 항체와의 반응성 확인을 위한 immunoblot 방법에서도 열처리에 따른 반응성의 차이는 나타나지 않았으며 콩 알레르기 환자의 혈청 역시 가열 처리에 따른 콩과의 반응성의 차이가 ELISA법을 이용한 측정에서 관찰되지 않았다. 이상의 결과로 부터 콩 알레르겐의 항원성 및 알레르기성은 가열처리에 의해 변화되지 않음을 확인할 수 있었다.

• 한국식품영양과학회지
• /
• 제28권2호
• /
• pp.277-284
• /
• 1999

To confirm the food quality of conventionally processed fish extracts, protein quality of boiled crucian carp(Carassius carassius) and bastard halibut(Paralichthys olivaceus) extracts and their residues were evaluated. For the both fish extracts, some of the essential amino acids were lowered significantly but two times more proline and glycine were detected in extracts than those in raw fish meats. Boiling(100oC, 5 hours) caused 1.8(crucian carp)~2.4(bastard halibut) times more total free amino acid contents in fish extracts as compared to those in original fish meats. Taurine, glutamic acid, proline, lysine, and ammonia were the predominant free amino acids released in fish extracts. In vitro digestibility of boiled fish extracts were lower at a level of 4~6% than those of raw fish meats. Fish extraction residue had a higher in vitro digestibility and had a 60% lower level of TI than that of original fish meats. 18(bastard halibut)~ 24%(crucian carp) of available lysine was reduced in boiled fish extracts but a remarkable variation was not noted between extracts and residues. PERs and NPRs of fish extracts were significantly lower than those of casein, while those values of extraction residue were slightly higher as compared to those of control(ANRC casein). In vivo apparent digestibility exhibited a similar trend to in vitro digestibility. Hematological properties in serum of rat fed with fish extracts and residue were not changed significantly but the serum cholesterol concentration were reduced in rats fed fish extraction residue comparing with those of control. These results suggest that body weight loss due to fish extracts may not affect physiological changes.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권9호
• /
• pp.1254-1260
• /
• 2003

The purpose of this study firstly was conducted to establish a radioimmunoassay (RIA) of estrone sulfate ($E_1S$), secondly to monitor the reproductive status of dairy cows using their urine samples. Urine and blood samples were collected in series within a day from four pregnant Holstein-friesian cows to evaluate the relationship between $E_1S$ levels in blood and urine with or without urinary creatinine basis. The urine was then collected biweekly from three cows in estrous and those artificially inseminated; collection from pregnant cows was made on a monthly basis. Results indicated that sensitivity for the $E_1S$ RIA was 5 pg/tube and the recovery rate was 100%. The daily urinary creatinine concentrations fluctuated within a day, but changes were slighter in midday, whereas the changes of concentrations of $E_1S$ in urine were relatively smaller. The concentrations of serum $E_1S$ during the estrous cycle were undetectable due to the limitation of assay, but the urinary $E_1S$ level could be measured with no obvious changes during the cycle. The urinary $E_1S$ levels increased remarkably around 7.7 to 8.3 ng/ml, 80 to 100 days after pregnancy but the serum $E_1S$ levels did not elevate until 120 to 150 days. The level of $E_1S$ increased gradually during pregnancy and eventually reached its peak before parturition at around 40 ng/ml and finally decreased to its basal level 2 days postparturition. During pregnancy, $E_1S$ concentrations of urine increased earlier than those in blood. The correlation coefficients between urinary and serum $E_1S$ concentration during pregnancy and postparturm were higher than those adjusted with creatinine (creatinine ratio). The concentrations of $E_1S$ in urine could be maintained unchanged for 8 days storing the samples in room temperature, which was extended to 8 days when the samples were pretreated by boiling for 30 minutes or treated with autoclave. In conclusion urinary $E_1S$ concentrations can be used directly for monitoring the pregnant status and fetal viability of dairy cows and can assist accurate confirmation of pregnancy in cows at least 80 to 100 days after insemination much earlier than by serum $E_1S$.