• IMMUNE NETWORK
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• v.11 no.6
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• pp.364-367
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• 2011

Background: Prostaglandins (PGs) play pathogenic and protective roles in inflammatory diseases. The novel concept of PGs as immune modulators is being documented by several investigators. By establishing an in vitro experimental model containing human follicular dendritic cell-like cells, HK cells, we reported that HK cells produce prostaglandin $E_2$ ($PGE_2$) and prostaglandin $I_2$ ($PGI_2$) and that these PGs regulate biological functions of T and B cells. Methods: To investigate the respective contribution of cyclooxygenase-1 (COX-1) and COX-2 to $PGE_2$ and $PGI_2$ production in HK cells, we performed siRNA technology to knock down COX enzymes and examined the effect on PG production. Results: Both $PGE_2$ and $PGI_2$ productions were almost completely inhibited by the depletion of COX-2. In contrast, COX-1 knockdown did not significantly affect PG production induced by lipopolysaccharide (LPS). Conclusion: The current results suggest that mPGES-1 and PGIS are coupled with COX-2 but not with COX-1 in human follicular dendritic cell (FDC) and may help understand the potential effects of selective COX inhibitors on the humoral immunity.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.257-265
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• 1996

Previously, we demonstrated that estradiol (E$_2$) was produced by medium sized follicles of Rana nigromaculata and Rana dybowskii in vitro. Present experiments were carried out to determine when the growing follicles have obtained the ability to produce E$_2$. Follicles in different growth stages were isolated and cultured for 6 hours in the presence or absence of frog pituitary homogenates (FPH, 0.1 pituitary/2m1) or various steroid precursors (200 ng/2m1). levels of progesterone (P$_4$), 17 -hydroxyprogesterone (17$\alpha$-OHP), androstenedione (AD), testosterone Cr) or E$_2$ in the medium were measured by RIA. The smallest follicles failed to produce steroids, whereas the smaller follicles produced considerable amounts of steroids (211 pg/follicle), and the medium sized follicles produced a large amounts of steroids (1653 pg/follicle) in response to FPH. Addition of pregnenolone (P5) resulted in a marked increase in P$_4$ but not in other steroids by the smallest follicles whereas the treatment resulted in a marked increase in P$_4$, 17$\alpha$-OHP, AD, T and E$_2$by the smaller and medium follicles. When the amounts of steroids are calculated on the basis of unit surface area (pg/mm$_2$), the ability of the smallest follicles to produce P$_4$ from P5 was similar to those of smaller and medium sized follicles. However, the smallest follicles failed to metabolize P$_4$ to other steroids whereas the smaller and medium follicles did. Taken together, the data suggest that the smallest follicles do not response to FPH in terms of steroid production but they have capacity to convert P5 to P$_4$ and that the smaller follicles have potential to produce E$_2$ although much less efficient than medium sized follicles.

• Journal of Pharmaceutical Investigation
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• v.29 no.4
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• pp.337-341
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• 1999

We have studied the stability and transdennal flux of prostaglandin $E_1\;(PGE_1)$ from various donor solutions through hairless mouse skin. Stability in HEPES buffer or in propylene glycol (PG) solution where enhancer (oleic acid (OA), propylene glycol monolaurate (PGML), transcutol (TC), ethanol (EtOH))s dissolved was investigated. $$PGE_1 was not stable in HEPES buffer. The concentration of $$PGE_1 decreased continuously for 7 days, and the degradation rate constant was $0.0028\;h^{-1}$, assuming first order reaction. The effect of current or penetration enhancer on the degradation was minimal. Percutaneous transport from HEPES buffer by passive or iontophoretic delivery without enhancer was close to nil. When OA or PGML was used together with PG, both passive and iontophoretic flux increased. PGML showed better enhancing effect than OA. Flux by cathodal delivery was about 2 times larger than that by passive delivery. Flux by anodal delivery was lower than that by passive delivery. TC and EtOH also increased the transdermal flux, but the effect was not as good as that observed when OA or PGML was used. These stability and flux data provide important information on how to formulate the patch, which will be the next step of this work, and on the polarity of current to use during iontophoresis.

• Journal of People, Plants, and Environment
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• v.21 no.6
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• pp.485-492
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• 2018

This study aimed to develop and validate highly sensitive determination method of a prostaglandin ($PGE_1$, OP 1206) in human plasma by LC-MS/MS using column switching. Plasma stored at $-30^{\circ}C$ and treated with methanol effectively inhibited interferences synthesized post-sampling. Samples were added with internal standard and were separated by reversed-phase HPLC with a cycle time of 30min. The method was selective for OP 1206 and the regression models, based on internal standard, were linear across the concentration range 0.5-50 pg/mL with the limit of quantification of 0.5 pg/mL (limit of quantitation, LOQ) for OP 1206. The calibration curve of OP 1206 standards spiked in five individual plasma samples was linear ($r^2=0.9999$). Accuracy and precision at the concentrations of 0.5, 1.5, 5.0 and 40 pg/mL, and at the lower LOQ of 0.5 pg/mL were excellent at 20%. OP120 < 6 was stable in plasma samples for at least 24 hours at room temperature, 24 hours frozen at $-70^{\circ}C$, 24 hours in an auto sampler at $6^{\circ}C$, and for two freeze/unfreezing cycles. The validated determination method successfully quantified the concentrations of OP 1206 in plasma samples from simulated administrating a single $5{\mu}g$ OP 1206 formulation. Thus, this novel LC-MS/MS technique for drug separation, detection and quantitation is expected to become the standard highly-sensitive detection method in bioanalysis and to be applied to many low dose pharmaceutical products.

• International Journal of Oral Biology
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• v.35 no.3
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• pp.113-119
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• 2010

Porphyromonas (P.) gingivalis lipopolysaccharide (Pg LPS) is the major pathogenic component of periodontal disease. In this study, we have attempted to determine the expression profiles of the signal transduction pathway genes induced by Pg LPS in comparison with Escherichia (E.) coli LPS (Ec LPS). DC2.4 cells were treated for two hours with $1\;{\mu}g/ml$ of Pg LPS or $0.5\;{\mu}g/ml$ of Ec LPS. The total RNA from these cells was then isolated and reverse-transcribed. Gene expression profiles were then analyzed with a signal transduction pathway finder GEArray Q series kit and significant changes in expression were confirmed by real-time PCR. The microarray results indicated that several genes, including Tnfrsf10b, Vcam1, Scyb9, Trim25, Klk6, and Stra6 were upregulated in the DC2.4 cells in response to Pg LPS treatment, but were downregulated or unaffected by Ec LPS. Realtime PCR revealed that the expression of Trim25, Scyb9 and Tnfrsf10b was increased over the untreated control. Notably, Trim25 and Tnfrsf10b were more strongly induced by Pg LPS than by Ec LPS. These results provide greater insight into the signal transduction pathways that are altered by P. gingivalis LPS.

• Journal of Electrochemical Science and Technology
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• v.7 no.1
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• pp.68-75
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• 2016

In this study a new, simple, precise, accurate and economic electrochemical method was developed and validated for the voltammetric determination of zolpidem (ZP) using disposable pencil graphite (PG) electrode. The anodic oxidation of ZP on the surface of the PG electrode was examined in a britton robinson (BR) buffer. Square wave and cyclic voltammetry were used as electrochemical techniques in the potential range of 0-1.2 V in the pH 8 BR buffer. In cyclic voltammetry studies, the diffusion coefficient of ZP oxidation was found to be 3.6×10^-6 cm² s^-1. On the other hand, the ZP has shown a well-defined irreversible anodic peak at 0.98 V in the square wave voltammetry mode. The PG electrode, primarily being graphite which has a large active surface area gives rise to increasing peak current with respect to ZP electrooxidation. PG electrode showed an electrocatalytic effect in anodic oxidation of ZP. A linear relationship between catalytic current response and ZP concentration was obtained over a concentration range of 10-30 μM with R.S.D. values ranging from 0.29-3.89. Limits of detection and quantitation were found to be 1 and 3 μM, respectively. Finally, the PG electrode was successfully used to determine ZP in standard and tablet dosage forms with a mean recovery of 100.69 %.

• Tuberculosis and Respiratory Diseases
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• v.49 no.2
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• pp.217-224
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• 2000

Background : It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-$\alpha$ and IL-1$\beta$. However, there is an alteration in the macrophages' responsiveness when they are challenged with repeated bouts of endotoxin, termed "endotoxin tolerance" which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. Methods : Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE II score. Peripheral blood monocytes were isolated from the patients and diluted to $1{\times}10^5$ well. After stimulation with endotoxin (LPS of E. coli O114 : B4, 100 ng/ml), they were incubated at $37^{\circ}C$ in 5% $CO_2$ incubator for 24 hours. Supernatant was collected for the measurement of TNF-$\alpha$ and IL-1$\beta$ with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. Results : The APACHE II score (mean$\pm$SD) of the patients at the time of blood sampling was 12.2$\pm$5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods (10 cases), gram positive cocci (6 cases) with two cases of mixed infection. Serum TNF-$\alpha$ could be measured in 4 cases with 29.9$\pm$27.7 pg/ml. Serum IL-1$\beta$was measurable in only one patient. The TNF-$\alpha$ level of supernatant of cultured peripheral blood monocytes was 2,703$\pm$2,066 pg/ml in patients and 2,102$\pm$1914 pg/ml in controls. The IL-1$\beta$level of supernatant was 884$\pm$1,050 pg/ml in patients and 575$\pm$558 pg/ml in controls. There was no difference of TNF-$\alpha$ and IL-1$\beta$ level between patients and controls. Conclusion : We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1566-1573
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• 2006

The study was conducted on 30 true acyclic Sahiwal cows (15 cows, ${\geq}90$ days postpartum; 15 postpubertal heifers, ${\geq}30$ months of age) and a similar 20 untreated controls (10 cows, 10 heifers). An 'Eazi' breed Controlled Internal Drug Release (CIDR) device (containing 1.38 g progesterone) was inserted intravaginally for 7 days (days 0 to 7) followed by 500 IU eCG i.m. at CIDR removal in all the treated animals. Heifers also received 5 mg oestradiol valerate i.m at CIDR insertion. The reproductive performance of these animals was recorded in terms of oestrus induction response, conception and pregnancy rates. Plasma progesterone ($P_4$) and oestradiol-$17{\beta}$ ($E_2$) profiles of 4 representative animals from each treatment group before, during and after CIDR treatment were also monitored. An oestrus induction response of 100% was observed in treated cows and heifers. The majority of cows (53.3%) and heifers (60%) were induced to oestrus within 24-36 and 36-48 h, respectively after CIDR withdrawal; with mean intervals of $44{\pm}3.18$ and $48{\pm}2.35h$, respectively. The conception rate at induced oestrus was higher in cows (40%) than heifers (20%). The final pregnancy rates after 2 subsequent oestruses were 80 and 60% in cows and heifers, respectively (overall 70% for all treated animals). In comparison, only 10% of control animals (2 cows only, 2/20) showed oestrus and become pregnant (10%) during theentire study period. The pretreatment (day 0) mean plasma P4 levels were statistically (p>0.05) similar in cows and heifers ($0.40{\pm}0.04$ and $0.49{\pm}0.11ng/ml$, respectively). The peak $P_4$ levels were observed on day 1 in cows ($13.94{\pm}1.41ng/ml$) and day 2 in heifers ($19.15{\pm}3.30ng/ml$) with a progressive decline up to the day of CIDR withdrawal ($3.35{\pm}0.92$ and $8.79{\pm}1.71ng/ml$, respectively). Mean $P_4$ levels on day 9 and 10 in cows and heifers did not differ significantly from their respective day 0 values and the lowest values were recorded on day 10 both in cows and heifers ($0.13{\pm}0.03$ and $0.14{\pm}0.02ng/ml$, respectively). Wide variations in individual pretreatment $E_2$ levels were observed both in the cows (range = 4-26, mean = $13.00{\pm}4.65pg/ml$) and heifers (range = 10-14, mean = $11.50{\pm}0.96pg/ml$). Thereafter also, $E_2$ levels in cows showed variation and reached a peak level ($53.50{\pm}2.99pg/ml$) on day 8. In heifers, peak mean $E_2$ level ($111.25{\pm}39.81pg/ml$) was recorded on day 1, followed by a non-significant decline on day 2, a significant fall on day 6 and a non-significant increase on day 9 and 10. However, mean $E_2$ levels on days 7 (p<0.05), 8 and 9 (p<0.01) were significantly higher in cows compared to heifers. The post-CIDR withdrawal mean highest $P_4$ and lowest $E_2$ levels coincided with the period when the majority of animals were induced to oestrus. CIDR and eCG treatment resulted in effective induction of oestrus with satisfactory pregnancy rates in true acyclic Sahiwal cows and heifers.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.1
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• pp.51-59
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• 1996

Controlled Ovarian hyperstimulation(COH) is generally used to obtain synchronous high quality oocytes in in vitro fertilization-embryo transfer(IVF-ET). Many investigators have studied the relationship between serum hormone levels and outcomes of IVF-ET because there is no accurate estimation method of oocyte quality. Early premature luteinization of follicles before oocyte retrieval is the most troublesome problem in COH for IVF-ET. Gonadotropin-releasing hormone agonists(GnRH-a) are used as adjuncts with gonadotropins for COH in patients undergoing in IVF. The possible benefits of GnRH-a pretreatment include improving oocyte quality, allowing a more synchronous cohort of follicles to be recruited, and preventing premature lueinization hormone surges. In COH of IVF cycles, we investigated whether an elevated progesterone(P4) level on the day of human chorionic gonadotropin(hCG) administration indicates premature luteinization and is associated with a lower fertilization rate. Many investigators have studied that the lower fertilization rates seen in patients with elevated P4 levels might result from an adverse effect of P4 on the oocytes. We hypothesizes that serum P4 levels around the day of hCG may be helpful prediction of out come in IVF-ET cycles. Success rates after COH of IVF-ET cycles are dependent upon many variable factors. Follicular factors including the number of follicles, follicular diameters and especially serum estradiol(E2) levels as an indirect measurement of follicular function and guality have been thought to influence the outcomes of IVF-ET. To assess whether serum P4 and E2 levels affect the fertilization and pregnancy rate, we reviewed the stimulation cycles of 113 patients (119 cycles) undergoing IVF-ET with short protocol with GnRH-a, from March 1993 to August 1994 retrospectively. The serum P4 and E2 levels were compared on the day of hCG in the pregnant group, 45 patients(47 cycles) and in the non-pregnant group, 68 patients (72 cycles) respectively. The serum E2 level in non-pregnant group was $1367{\pm}875.8$ pg/ml which was significantly lower than that of pregnant group, $1643{\pm}987.9$ pg/ml( p< 0.01 ). And the serum P4 level in non-pregnant group was $2.1{\pm}1.4$ ng/ml which was significantly higher than that of pregnant group, $1.0{\pm}0.7$ ng/ml( p< 0.001 ). The fertilization rate was $61.3{\pm}21.3%$ in pregnant group which was higher than that of non-pregnant group, $41.1{\pm}20.2%$ (p< 0.01). We suggest that the serum levels of P4 and E2 on the day of hCG administration are additional parameters that predict the outcomes of IVF-ET cycles.

• Korean Journal of Animal Reproduction
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• v.14 no.3
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• pp.175-182
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• 1990

Behavioural estrus and short estrous cycles were observed and serum concentrations of estradiol-17$\beta$(E2) before and after of estrous were measured following superovulation treatments in 30 pluriparous Korean native goats. The goats were divided into 2 groups. Fifteen goats were injected IM with 1,000IU PMSG on Day 12 of the estrous cycle followed by 10mg PGF2$\alpha$ 48h later(P4＋PMSG), and the other 15 goats were injected IM with 10mg progesterone(P4)in oil once daily for 10d beginning at any days of estrous cycle followed by 1,000IU PMSG and 10mg PGF2$\alpha$ at the 8th day of progesterone treatment(P4＋PMSG group). After injection of PGF2$\alpha$, onset of standing estrus occurred in 12 of 15 goats(80.0%) at 50.0$\pm$7.7h and in 11 of 15 goats(73.3%) at 135.6$\pm$10.1h in PMSG and group and P4＋PMSG group, respectively. The mean interval from PGF2$\alpha$ injection to first estrus was significantly(P<0.01) earlier in PMSG group than in P4＋PMSG group. This result indicate that the delayed infusion of P4 in P4＋PMSG group caused the later exhibition of their estrous behaviors. However, duration fo frist estrus(31.5$\pm$2.6h vs 26.2$\pm$2.3h), length of estrous cycle(14.1$\pm$3.3d vs 16.6$\pm$3.8d) and percentage of short estrous cycle(50.0% vs 45.5%) were not different between PMSG and P4＋PMSG group. The mean concentration of serum E2 in 4 goats showing normal estrous cycle in P4＋PMSG group(PP-NEC) was higher than in 6 goats showing normal(P-NEC) or in 6 goats showing short estrous cycle(P-SEC) in PMSG group. The peak level of serum E2 was observed at the time of onset of standing estrus in PP-NEC(67.6pg/ml), 6h earlier in P-NEC(53.1pg/ml) and 6h later in P-SEC(52.3pg/ml) than the onset of standing estrus. The profiles of serum concentration of E2 during the period of peri-estrus was similar in the goats of PMSG or P4＋PMSG and also in the goats showing the subsequent estrous cycle of normal or short length.