• The Korean Journal of Nuclear Medicine Technology
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• v.14 no.2
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• pp.190-196
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• 2010

Purpose: This study was undertaken to investigate the feasibility of using automated liquid-handling systems equipped with reusable fixed tips in Radioimmunoassays and to demonstrate that the use of an automated pipetting instrument can decrease in the typical daily workload. Materials and Methods: The precisions of the automated pipettor and the manual pipettor were determined gravimetrically (n=30). A total of 30 specimens for HBs Ag were repeatedly pipetted (three replicates) with the automated pipettor and then retested. PSA samples were simultaneously pipetted with either the automated pipettor or the manual pipettor and then analyzed (n=40). Sample carryover test assessed for CA19-9, AFP and HCG when the automated pipettor was used. Pipetting speed of the automated pipettor and the manual pipettor were compared by evaluation of each workload. Results: The precisions coefficients of variation (CV) were 2.1% for the automated pipettor and 1.6% for manual pipettor. The mean cpm and CV for each group of replicates were 41,203 cpm and 3.7% for HBs Ag positive specimens, and 99 cpm and 7.9% for HBs Ag negative specimens, respectively. PSA results showed no significant differences between automated pipettor and manual pipettor (p=0.15, r=0.999). Carryover for CA19-9, AFP and HCG analytes was <0.1 ppm or below the assay limit of detection. Pipetting speed was significantly improved by using the automated instrument. Conclusion: There was no evidence that the use of an automated pipettor adversely affected any of the performance characteristics of the assay. Indeed, routine use of the Tecan automated pipettor has resulted in a decrease in the typical daily workload.

• Journal of Korean Neurosurgical Society
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• v.63 no.1
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• pp.56-68
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• 2020

Objective : To analyze the incidence and treatment trends of hemorrhagic stroke (HS), according to HS subtypes, using nationwide data in Korea from January 2008 to December 2016. Methods : We used data from the national health-claim database provided by the National Health Insurance Service for 2008-2016 using the International Classification of Diseases. The crude incidence and age-standardized incidence of each disease associated with HS, which included intracranial aneurysm (IA), hypertensive intracerebral hemorrhage (ICH), and arteriovenous malformation (AVM), were calculated and additional analysis was conducted according to age and sex. Changes in trends in treatment methods and number of treatments were analyzed for each cerebrovascular disease using the Korean Classification of Diseases procedure codes. Results : In 2016, the total number of newly diagnosed adult patients with HS was 24169, showing a decrease by 7.0% since 2008; the age-standardized incidence of HS was 46.2 per 100000 person-years. The age-standardized incidence of unruptured IA (UIA) in adults was 71.4 per 100000 person-years-increased by 2.6-fold since 2008-while that of ruptured IA (RIA) was 12.6 per 100000 person-years, which had decreased at a rate of 20.3% since 2008. The number of coil embolization (CE) for UIA increased by 3.4-fold over 9 years and exceeded that of clipping since 2008. With respect to RIA, CE increased by 2.0-fold over 9 years and exceeded that of clipping from 2014. As for spontaneous ICH in adults, the age-standardized incidence was 31.3 per 100000 person-years in 2016-decreased by 34.7% since 2008-and 14.6% of patients diagnosed with ICH were treated in 2016, which was not significantly different from the proportion of patients treated since 2008. The age-standardized incidence of unruptured AVM (UAVM) was 2.0 per 100000 person-year in 2016, while that of ruptured AVM (RAVM) was 2.4 per 100000 person-years in 2016, showing a decreasing rate of 17.2% from 2008. The total number of treated patients with AVM declined since 2014. Conclusion : In Korea, age-related cerebral vascular diseases, such as RIA, ICH, and RAVM, demonstrated a declining trend in age-standardized incidence; meanwhile, UIA and UAVM demonstrated an increased trend in both crude incidence and age-standardized incidence for 9 years. The increase in the elderly population, management of hypertension, and development of diagnostic and endovascular techniques appear to have influenced this trend.

• Development and Reproduction
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• v.4 no.2
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• pp.237-242
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• 2000

Growth hormone releasing hormone (GHRH), the major hypothalamic stimulus of GH secretion from the anterior pituitary gland, has been found to be present in several extrahypothalamic sites including placenta testis, ovary and anterior pituitary gland. The present study was performed to elucidate the role of pituitary GHRH on proliferation of cells derived from rat anterior pituitary gland. The GHRH content of pituitary tissue, cultured pituitary cells, and the conditioned media was evaluated by radioimmunoassay (RIA). Primary cultures of pituitary cells derived from adult rats were prepared by enzymatic dispersion. Significant amounts of GHRH-like molecules were detected in both pituitary tissue and cell cultures by GHRH RIA. Competition curves with increasing amounts of tissue extracts and conditioned media were parallel with those of standard peptide, indicating that the pituitary GHRH-like material is similar to authentic GHRH. To analyze specific cell types responsible for producing GHRH in anteroior pituitary, cell fractionation technique combined with GHRH RIA was performed. In cell fractionation experiment, the highest level of GHRH content was found in gonadotrope enriched-fraction and followed by somatotrope-, lactotrope- and thyrotrope-fraction. Treatment of pituitary cells with GHRH resulted in a dose-dependent increase in [$^3$H] thymidine incorporation. The mitogenic effect of GHRH could be mediated by typical oncogenic activation since the GHRH induced transient increase in c-fos mRNA levels with peak response at 30 minutes. The present study demonstrated that i) the pituitary GHRH expressed in the rat anterior pituitary gland can be secreted, ii) among the various cell types, gonadotropes and somatotorpes are the major GHRH source, and iii) the GHRH treatment increased the [$^3$H] thymidine incorporation and c-fos transcriptional activity in the pituitary cell culture. These findings suggested that GHRH could participated in the paracrine and/or autocrine regulation of cell proliferation, as well as promoting growth hormone secretion.

• Development and Reproduction
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• v.4 no.2
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• pp.231-236
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• 2000

Recent studies have clearly shown that the expression of genes for gonadotropin-releasing hormone (GnRH) and its receptor in the rat reproductive organs including ovary, testis, placenta uterus and mammary gland. Moreover, luteinizing hormone (LH) classically known to be a main target product of GnRH in anterior pituitary has been found in rat gonads. These findings suggested the presence of local circuit composed of GnRH and LH in the rat gonads. The present study was undertaken to elucidate whether the genes for LH and its receptor are expressed in rat mammary gland. Expression of LH and its receptor genes in the rat mammary gland was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) and specific LH radioimmunoassay (RIA). The LH${\beta}$ transcripts in the mammary gland from cycling rats contained the pituitary type of LH${\beta}$ exons 1~3 encoding the entire LH${\beta}$ polypeptide but lacked the rat testis-specific LH${\beta}$ exon(s). Presence of ${\alpha}$ -subunit transcripts in the rat mammary gland were determined by RT-PCR. The cDNA fragments encoding exons 2~7 of rat LH receptor transcripts were amplified in both rat ovary and mammary gland samples. We could detect the GnRH expression in mammary gland from cycling virgin rats, and this result disagreed with previous report that mammary GnRH expression is occured in lactating rats only. Considerable amounts of immunoreactive LH molecules with good RIA parallelism in standard curve were detected in crude extracts from the rat mammary gland, indicating that the immunoreactive LH materials in the gland might be identical to authentic pituitary LH. To our knowledge, the present study demonstrated for the first time the expression of LH subunits and LH receptor in the rat mammary gland. Our findings suggested that the mammary gland might be the novel source and target of LH and the mammary LH could be act as a local regulator with auto-and/or paracrine manner under the regulation of local GnRH.

• Journal of Life Science
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• v.8 no.6
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• pp.695-701
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• 1998

This study was carried out to investigate levels of the components of microsomal mixed function oxidase (MFO) system and activities of the hepatic microsomal cytochrome P45O (P45O)-dependent monooxygenases of grass snake (Natrix tigrina Lateralis) and to compare with those of rat. The levels of P45O and cytochrome b$_{5}$, (b$_{5}$) of snake were much lower than those in rat. NADPH-cytochrome c reductase activity in the snake was also only 40% of that in the rat. Activities of 7-ethoxycoumarin 0-deethylase (ECOD) and benzphetamine N-demethylase (BPDM) of snake hepatic microsomes, when compared with those of rat, were markedly low. But, aryl hydrocarbon hydroxylase (AHH) and testosterone hydroxylase (TSH) activities were nearly the same or higher than those of the rat. Of the P45O-dependent TSHs measured, 7$\alpha$-hydroxylase activity was the highest in snake, whereas, 6$\beta$-hydroxylase activity was the highest in rat. However, stereoselectivity of the enzyme from the snake to C2 and C6 positions of testoste-rone was the same as rat. The result of radioimmunoassay (RIA) for the identification of five P45O isozymes with MAbs shows that relatively high content of ethanol-inducible P45O isozyme, CYP2El, exists in the rat, whereas MC-inducible P45O isozyme, CYP2A1/1A2, does in the snake. From the analyses of SDS-PAGE and RIA of partially pu-rified P45O, we suggest the possibility of the presence of a certain P45O isozyme(s) in hepatic microsomes of snake different from those of rat.

• The Korean Journal of Nuclear Medicine Technology
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• v.21 no.2
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• pp.28-30
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• 2017

Purpose Anti-Glutamic acid decarboxylase antibody test (GAD Ab) has been used as a predictor of type 1 diabetes. GAD Ab has also been shown to be highly potent in cerebrospinal fluid (CSF) of patients with suspected diabetic peripheral neuropathy. Recently, it has been known that clinical significance of GAD Ab using CSF is useful for the neurological disorders. However, the reference value of anti-GAD Ab has been provided only for serum. In this experiment, we estimated the reference value of anti-GAD antibody for CSF in neurological patients. Materials and Methods A total of 211 neurological patients were enrolled. Serum and CSF were analyzed by radioimmunoassay (RIA) using commercial RIA anti-GAD Ab kit (RSR, London, United Kingdom). Normal saline was used as the normal CSF control because CSF is most similar to 0.9% normal saline. Results The mean value of normal CSF control was 1.97 U/mL, and two standard deviations (2SD) was 1.44 U/mL. Based on this data, the expected reference range of CSF could be estimated from 0.54 U/mL to 3.40 U/mL Conclusion The reference range of normal CSF control using normal saline obtained with Hoffmann's method. However, there will be a need to validate the CSF reference values using human normal CSF.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.4
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• pp.460-466
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• 2004

This study was undertaken with the aim to establish a reliable radioimmunoassay (RIA) system for urinary pregnanediol glucuronide (PdG) and to employ it for monitoring the reproductive status of dairy cows. Urine and blood samples were collected from the Holstein cows both pregnant and non-pregnant. The samples were then investigated for evaluating the relationship between progesterone ($P_{4}$) in blood and PdG in urine adjusted with or without urinary creatinine basis. Biweekly urine collection was employed for three cows in estrous and those artificially inseminated, while urine from pregnant cows was collected on a monthly basis. P_{4}$ and PdG levels were measured by enzymeimmunoassay (EIA) and RIA techniques, respectively. Our results indicated the sensitivity of PdG for RIA being 35 pg/tube and the recovery rate of 100%. Urinary creatinine concentrations also fluctuated within a day, but change at midday was not noteworthy. Regardless of the time of urination the change in concentrations of PdG was relatively smaller and did not vary significantly. The urinary PdG concentration showed periodic changes as that with serum P_{4}$ levels during the cow's estrus cycle. The correlation coefficient rose when creatinine level in urine was adjusted but the change was also not significant. The concentrations of PdG during the luteal phase were detected between 8.2 and 17.4 ng/ml, three to five times higher than that in the follicular phase. The concentration of PdG from pregnant cows (21 days after conception) was three to four times higher than in the nonpregnant cows. Our finding suggests that the determination of urinary PdG could be reliably employed for early pregnancy detection. The urinary PdG level continued to raise until 30 days pre-partum while the concentration reached its peak at 30 ng/ml, after which it started to fall 18 to 30 days before parturition and finally fell to its nadir value one week after parturition. As the correlation coefficient between the urinary PdG and serum P_{4}$ was higher than that corrected by urinary creatinine it can be suggested that the adjustment is not needed. The concentrations of urinary PdG could be maintained stably for 2 days in urine samples stored at room temperature and extended to 8 days when the samples were pretreated by boiling for 30 minutes. In conclusion urinary PdG concentration even without the need for creatinine basis adjustment can be used directly for monitoring the reproductive status of dairy cows.

• Development and Reproduction
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• v.5 no.1
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• pp.59-71
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• 2001

Since the first radioimmunoassay (RIA) was developed in 1970s, many conventional RIAs and non-isotopic immunoassays (NIA) had been developed in which the degree of precision, accuracy, specificity and practicability have progressively increased. Recently ultrasensitive assay method at femtogram to determine testosterone in serum, saliva and feces is required for the study of sexual dysfunctions in male and female, monitoring the psychological stress and conditions, aging process such as menopause and partial androgen deficiency in aging male, the hormonal changes of small experimental animals etc. This review discussed the recentd evelopments of steroid assay methods, based upon the testosterone assay results of authors far 20 years, and the problems associated the assay set-up, the characterizations and applications of the established procedures, and desifls of assay, reliablity criteria, and the practical aspects of assay set-up and application, based upon the data of the authors. The present study demonstrates the general problems methods to be consider in order to set up the highly sensitive assay methods and to increase the assay quality and the necessity of assay quality control program. To improve the assay quality of each laboratory and to compare the assay results in homeland, the national QC programs should be organized.

• Korean Journal of Animal Reproduction
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• v.9 no.2
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• pp.133-139
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• 1985

An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.

• 대한핵의학회:학술대회논문집
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• 1999.11a
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• pp.26.2-26.2
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• 1999