A 1 : 1 mixture of acriflavine (ACF; CAS 8063-24-9) and guanosine is currently being evaluated as a possible antitumor agent in preclinical studies. Guanosine is known to potentiate the anticancer activity of some compounds. However, the distributions of trypaflavine (TRF) or proflavine (PRF) have not been investigated in mammals. We, therefore, investigated the distribution of TRF and PRF after i.m. administration of the combination mixture (ACF and guanosine) at a dose of 30 mg/kg ACF in rats. to analyze TRF and PRF levels in biological samples, we used an HPLC-based method. The calibration curves for TRF and PRF in the samples were linear over the concenration range of $0.05{\sim}200\;{\mu}g/ml$. The intra- and inter-day assay accuracies of this method were within ${\pm}15\%$ of norminal values and the precision did not exceed $15\%$ of relative standard diviation. The lower limits of quantitation were 50 ng/ml for both TRF and PRF. The distribution of TRF or PRF was determined by 48 h after i.m. administration of the combination mixture at a dose of 30 mg/kg ACF. TRF and PRF were distributed as the following order; kidney>lung>liver>small intestine>muscle. Of the various tissues, TRF and PRF were mainly distributed to the kidney and lung. The concentrations of TRF or PRF in the tissues 24 h after i.m. administration decreased to undetectable levels. The concentrations of TRF or PRF in the blood cells were comparable to those for the plasma. However, the concentrations of TRF or PRF in the both plasma and blood cells 12 h after i.m. administration were not detected. The number of the platelets in the 1 ml of the blood was calculated to be $0.183{\times}10^8/ml$ of blood. The PRF concentration in platelets was higher than that of TRF at initial times after i.m. administration of the combination mixture. However, both the TRF and PRF concentrations in the plateles 24 h after i.m. administration of the combination mixture were below the quantifiable limit. In conclusion, the concentrations of TRF or PRF in the various tissues, plasma, blood cells, and plateles decreased to undetectable levels 24 h after i.m. administration of the combination mixture at a dose of 30 mg/kg ACF.
Background : Endothelin(ET) is a very potent vasoconstrictive peptide produced by endothelial cells of pulmonary artery. The endothelin level was increased in plasma of primary pulmonary hypertension and acute pulmonary thromboembolism and it was suggested that the endothelin might do a critical role in the cardiopulmonary dysfunction in these two conditions. But the exact mechanism of increase of ET has not been known. In these two conditions, platelet activation and thrombosis are the main pathophysiologic findings. So there is a possibility that the platelet might stimulate endothelin secretion from endothelial cells. Therefore, we performed this study to evaluate the role of platelet and its mediators on endothelin production in bovine pulmonary artery endothelial(BPAE) cells. Method : Bovine pulmonary artery endothelial cells, ATCC certified cell line 209, were cultured and treated with human platelets($10^6{\sim}10^8/ml$), thrombin (0.1~10u/ml), TGF-${\beta}1$(1~100uM), serotonin(1~100uM), and endotoxin(1ug/ml) in a final volume of 500ul for 18 hours. Levels of ir(immunoreactive)-ET in each conditioned medium were measured by a radioimmunoassay specific for ET. Result : The increase of ir-ET levels was platelet number and time dependent over 18 hours. When washed human platelets were added($10^8/ml$), the ir-ET levels were significantly higher than that of control(p<0.05) at 8 and 18 hours after culture. Subthreshold concentration of platelets($10^7/ml$) coincubated with endotoxin(1ug/ml) or subthreshold dose of thrombin(0.1u/ml) stimulated ir-ET secretion from BPAE cells significantly(p<0.05) compared with control. Thrombin(1ug/ml, 10ug/ml) and TGF-${\beta}1$(100pM, 1000pM) significantly increased ir-ET secretion from BP AE cells(p<0.05) compared with control, but serotoin(1~100uM) and endotoxin(1ug/ml) did not stimulate the ir-ET secretion. Conclusions : Platelets stimulate endothelin secretion from bovine pulmonary artery endothelial cells. The mechanism of increase of endothelin secretion seems to be a stimulation by platelet itself or by mediators, such as TGF-${\beta}1$, secreted from activated platelets. And, in this study, the priming effect of platelets on endothelin secretion from BPAE cells could be another possibility.
We investigated the antiplatelet effect of a newly synthesized guanidine derivative KR-32558, a sodium-hydrogen exchanger-1 (NHE-1) inhibitor, together with the elucidation of the possible mechanisms of action. KR-32558 concentration -dependently inhibited the aggregation of washed rabbit platelets induced by collagen (10 ${\mu}g/ml$) with an $IC_{50}$ value of 85.9 ${\mu}M$, but with much weaker potency against aggregation induced by thapsigargin (0.5 ${\mu}M$) or A23187 (5 ${\mu}M$). And had no effect on platelet aggregation induced by arachidonic acid (100 ${\mu}M$), thrombin (0.05 U/ml) and U46619 (1 ${\mu}M$) up to 100 ${\mu}M$. KR-32558 completely inhibited the $[Ca^{2+}]_i$ mobilization induced by collagen at concentration of 100${\mu}iM$. Taken together, these observation suggest that KR-32558 selectively inhibited collagen-mediated platelet aggregation by blocking the cytoplasmic calcium mobilization in addition to NHE-1 inhibition.
Sweet potato has low regeneration capacity, which is a serious obstacle for the fruitful production of transgenic plants. Simple and rapid regeneration method from storage root explants of purple sweet potato (Ipomoea batatas L.) was investigated. The embryogenic callus was observed from 4 cultivars and its highest rate was induced at 1 μM 2,4-D after 5 weeks of culture. Result revealed that a low concentration of 2,4-D and low light intensity was important factors for embryogenic callus formation. After subculture on medium with 5 μM ABA for 4 days, subsequently, occurred the regeneration of shoots within 4 weeks when these embryogenic callus was transferred onto the MS hormone free medium. Regenerated shoots were developed into platelets, and grown normal plants in the greenhouse. We developed a simple and quickly protocol to regenerate plantlets in storage root explants of purple sweet potato. This regeneration system will facilitate tissue culture and gene transfer research of purple sweet potato.
These studies were conducted about the effects of Ferric oxide in the aspect of the basic pharmacology through experimental animals with the investigation of its clinical effects on oriental medical literatures of Ferric oxide. The results were as follows: 1. The analgenic effects induced by acetic acid were significant. 2. The anticonvulsive effects induced by ECT unit were significant. 3. The sedative effects in Rotor rod method were significant. 4. The number of platelets was increased but not significantly. 5. Prothrombin time was decreased significantly. 6. Fibrinogen was increased but not significantly. 7. The degree of concentration of FDP was decreased significantly. According to the above results, it is considered that Ferric oxide retains effectiveness on the treatment of cerebral stroke.
Kim, Min Kyoung;Song, Haengseok;Lyu, Sang Woo;Lee, Woo Sik
Clinical and Experimental Reproductive Medicine
/
v.49
no.3
/
pp.168-174
/
2022
Refractory thin endometrium and recurrent implantation failure are among the most challenging infertility-related factors hindering successful pregnancy. Several adjuvant therapies have been investigated to increase endometrial thickness and the pregnancy rate, but the treatment effect is still minimal, and for many patients, these treatment methods can be quite costly and difficult to approach. Platelet-rich plasma (PRP) is an autologous concentration of platelets in plasma and has recently been elucidated as a better treatment option for these patients. PRP is rich in cytokines and growth factors, which are suggested to exert a regenerative effect at the level of the injured tissue. Another advantage of PRP is that it is easily obtained from the patient's own blood. We aimed to review the recent findings of PRP therapy used for patients with refractory thin endometrium and recurrent implantation failure.
Gilson Khang;Park, Myoung-Kyu;Jong M. Rhee;Lee, Sang-Jin;Lee, Hai-Bang;Yasuhiko Iwasaki;Nobuo Nakabayashi;Kazuhiko Ishihara
Macromolecular Research
/
v.9
no.2
/
pp.107-115
/
2001
Poly(L-lactide-co-glycolide)(PLGA) was blended with poly[$\omega$-methacryloyloxyethyl phospho-rylcholine-co-ethylhexylmethacrylate (PMEH)] (PLGA/PMEH) to endow with new functionality i.e., to improve the cell-, tissue- and blood-compatibility. The characteristics of surface properties were investigated by measurement of contact angle goniometer, Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and electron spectroscopy for chemical analysis (ESCA). NIH/3T3 fibroblast and bovine aortic endothelial cell were cultured on control and PLGA/PMEH surfaces for the evaluation of ceil attachment and proliferation in terms of surface functionality such as the concentration of phosphoryl-choline. Also, the behavior of platelet adhesion on PLGA/PMEH was observed in terms of the surface functionality. The contact angles on control and PLGA/PMEH surfaces decreased with increasing PMEH content from 75$^{\circ}$ to about 43$^{\circ}$. It was observed from the FTIR-ATR spectra that phosphorylcholine groups are gradually increased with increasing blended amount of MPC. The experimental P percent values from ESCA analysis were more 3.28∼7.4 times than that of the theoretical P percent for each blend films. These results clearly indicated that the MPC units were concentrated on the surface of PLGA/PMEH blend. The control and PLGA/PMEH films with 0.5 to 10.0 wt% concentration of PMEH were used to evaluate cell adhesion and growth in terms of phosphorylcholine functionality and wettability. Cell adhesion and growth on PLGA/PMEH surfaces were less active than those of control and both cell number decreased with increasing PMEH contents without the effect of surface wettability. It can be explained that the fibronectin adsorption decreased with an increase in the surface density of phosphorylcholine functional group. One can conclude the amount of the protein adsorption and the adhesion number of cells can be controlled and nonspecifically reduced by the introduction with phosphorylcholine group. Morphology of the adhered platelets on the PLGA/PMEH surface showed lower activating than control and the number of adhered platelets on the PLGA/PMEH sample decreased with increasing the phosphorylcholine contents. The amount of fibrinogen adsorbed on the PLGA/PMEH surface demonstrated that the phospholipid polar group played an important role in reducing protein adsorption on the surface. In conclusion, this surface modification technique might be effectively used PLGA film and scaffolds for controlling the adhesion and growth of cell and tissue, furthermore, blood compatibility of the PLGA was improved by blending of the MPC polymer for the application of tissue engineering fields.
OA-8159, a new Phosphodiesterase (PDE) 5 inhibitor, has exhibited potent erectogenic potential in a penile erection test in rats and anesthetized dogs. In this study, we investigated the mechanism of its erectogenic activity by measuring the activity of OA-8159 against a various PDE isozymes and assessing cGMP and cAMP formation in a rabbit corpus cavernosum in vitro. DA-8159 inhibited the PDE 5 activity in rabbit and human platelets, which the $IC_{50}$ was 5.84$\pm$1.70 nM and 8.25$\pm$2.90 nM, respectively. The $IC_{50}$ of DA-8159 on PDE 1, PDE2, PDE 3 and PDE 6 were 870$\pm$57.4 nM, $101\pm$5 $\mu$M, 52.0$\pm$3.53 $\mu$M and 53.3$\pm$2.47 nM, respectively. This suggests that DA-8159 is a potent, highly selective, competitive inhibitor of PDE 5-catalyzed cGMP hydrolysis. The rates of cGMP hydrolysis catalyzed by human platelets-derived PDE 5 as a function of the cGMP concentration (5~100 nM) and two-fixed DA-8159 concentration (11.3 and 18.8 nM) were investigated in order to characterize the mode of PDE 5 inhibition by DA-8159. DA-8159 increased the apparent 4K_{m}$ value for cGMP hydrolysis but had no effect on the apparent $V_{max}$, indicating a competitive mode of inhibition. DA-8159 increased the cGMP concentrations in the rabbit corpus cavernosum dose dependently. In the presence of sodium nitroprusside (SNP), DA-8159 significantly sti\mulated the accu\mulation of cGMP when compared to the control level. This indicated that the enhancement of a penile erection by DA-8159 involved the relaxation of the cavernosal smooth \muscle by NO-sti\mulated cGMP accu\mulation. In conclusion, DA-8159 is a selective inhibitor of PDE 5-catalyzed cGMP hydrolysis and the enhancement of a penile erection by DA-8159 is mediated by the relaxation of the cavernosal smooth \muscle by the NO-sti\mulated cGMP accu\mulation.
Objective & Methods: The purpose of this study is to investigate the anti-oxidative and immuneregulative effects of electro-acupouncture(EA) at SP6(Sameumgyo) in aged rats. The author performed several experimental items including blood cell counts, blood chemistry, measurement of various oxidants and antioxidants in liver and spleen, analysis of various cytokines in spleen. The results are as follows. Results: 1. EA at SP6 significantly reduced the number of platelets in blood. 2. EA at SP6 significantly reduced NO concentration and significantly increased catalase activity in liver. 3. EA at SP6 significantly reduced NO concentration and significantly increased SOD activity, catalase activity and glutathione concentration in spleen. 4. EA at SP6 restored the increase of IL-4, IL-6 and the decrease of IFN-$\gamma$ in aged rat spleen. Conclusion: According to these results, it is postulated that EA at SP6 has an antioxidative effect through increasing the activities of antioxidative enzymes and inhibiting production of oxidized substances, as well as an immune regulative effect in aging process. In consequence, it is presumed that EA at SP6 may have an anti-aging effect.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.10
no.1
/
pp.1-38
/
1997
This study was performed to investigate the effects of Kamigyemyoungsan and each compositive herbs on the intravascular coagulation and subcutaneous hematoma. The intravascular coagulation was induced by the injection of endotoxin into the caudal vein of the rats. These rats were treated with endotoxin after administering orally extracts of Kamigyemyoungsan and each compositive herbs. Then the number of platelet, prothrombin time, the concentration of fibrinogen and FDP(fibrinogen degration product), hematocrit and RBC and WBC were measured. The subcutaneous hematoma was induced by the subcutaneous injection of the autologous whole blood to produce clotting in situ in rats. Then the extracts of Kamigyemyoungsan and each compositive herbs were administered orally. The lesions were then dissected and observed. The results were obtained as follows ; 1. The number of platelets of the trial groups was increased as compared with the control group, and revealed significance in sample Ⅰ. 2. The concentration of fibrinogen of the trial groups was increased as compared with the control group, and revealed significance in sample Ⅰ, sampleⅡ and sample Ⅴ. 3. The prothrombin time was shortened significantly in the trial groups, except sampleⅡ and sampleⅣ as compared with the control group. 4. The concentration of FDP decreased in the trial groups, and revealed significance in sample Ⅰ and sample Ⅵ as compared with the control group. 5. The hematocrit significantly increased in sample Ⅰ, sampleⅢ and sample Ⅵ as compared with the control group. 6. The number of RBC significantly increased in sample Ⅵ only as compared with the control group. 7. The number of WBC significantly increased in the trial groups, except sampleⅣ and sample Ⅵ as compared with the control group. 8. Histologically, the lesions in the trial groups showed significantly thinner fibroblastic neomembrane than the control group, except sampleⅢ. According to the above results, it is considered that Kamigyemyoungsan and each compositive herbs have inhibitive effects on the thrombosis and the fibroblastic neomembrane development. Therefore, it seems to be applicable to the diseases related to thrombosis and hematoma.
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