Development of Two Korean IACUC Guidance Documents to Foster Implementation of the Three Rs.

In Korea, Institutional Animal Care and Use Committees (IACUCs) have been legally required to apply the Three Rs principles (i.e. replacement, reduction and refinement) and undertake the ethical review of animal study protocols, since 2008. According to Korean law, each IACUC is required to appoint at least one lay member recommended by a non-governmental animal protection organisation, who participates in the ethical review process as part of this role. Despite the importance of the Three Rs and the ethical review process, limited information and practical resources are available for IACUC members in the Korean language, particularly for lay members who are inexperienced in animal experimentation. In January 2020, the Animal and Plant Quarantine Agency announced the funding for a six-month research project to develop guidance to assist IACUC members in carrying out effective and efficient protocol reviews in line with Korean legislative requirements. This funding was awarded for the production of two IACUC guidance documents - 'Guide for Animal Study Protocols' and 'Guide for the IACUC Lay Member' - which were published in December 2020. These guidance documents aim to foster the implementation of the Three Rs and provide practical resources for IACUC members, researchers and other relevant personnel. This paper describes the framework for animal use in Korea and the overall production of these two IACUC Guidance Documents.

Induction of liver transplant immune tolerance in an outbred rat strain model using tacrolimus.

BACKGROUND: Orthotopic liver transplantation is the only option for patients with end-stage liver disease and hepatocellular carcinoma. Post-transplant immunosuppressive therapy is important to prevent graft failure. We investigated the effectiveness of tacrolimus (FK506) and their mechanisms for liver transplant immune tolerance in an outbred rat LT model. RESULTS: To investigate the therapeutic effect of the FK506 on outbred rat LT model, FK506 and postoperative therapy were administered subcutaneously once or twice daily to transplanted rats. Histopathological and immunohistochemical analyses were conducted for all groups. The regulation of inflammatory cytokine signaling in the spleen was analyzed by flow cytometry. FK506 attenuated allograft rejection and increased survival in rat orthotopic liver transplantation models. The FK506-treated group had reduced serum levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase. Furthermore, FK506 decreased the expression of inflammatory cytokines and the activation of pathogenic Th1 and Th17 cells in the liver. CONCLUSIONS: Taken together, we revealed that FK506 ameliorated strong allograft rejection in outbred liver transplantation model by anti-inflammatory effect and inhibitory peroperty of pathogenic T cells.

Assessment of Risk Factors Affecting Refractive Outcomes after Phacovitrectomy for Epiretinal Membrane

Purpose@#To investigate factors associated with refractive outcomes after phacovitrectomy for epiretinal membrane (ERM). @*Methods@#Retrospective review of patients undergoing phacovitrectomy for ERM was done. The main outcome measure was predictive refraction error (PE), defined as observed refraction error – target refraction error, calculated by the SRK/T, Haigis, and SRK II formulae. PE was measured at postoperative 1, 3, and 6 months. Simple and multiple linear regression analysis were used to evaluate factors associated with PE. @*Results@#A total of 53 eyes of 53 patients were included. The mean PEs at postoperative 1, 3, and 6 months were all negative, implying myopic shift in all patients regardless of the intraocular lens formula used. Haigis formula showed the least myopic shift among the three formulae (p = 0.001, Friedman test). There was no significant difference in PE depending on preoperative central macular thickness (CMT) in subgroup analysis. On stepwise multiple linear regression analysis, ERM etiology (β = 0.759, p = 0.004, SRK/T formula; β = 0.733, p = 0.008, Haigis formula; β = 0.933, p < 0.001, SRK II formula), preoperative anterior chamber depth (β = –0.662, p = 0.013, Haigis formula; β = –0.747, p = 0.003, SRK II formula), and decrease of CMT (β = –0.003, p = 0.025, SRK/T formula) were significantly associated with PE at postoperative 6 months. @*Conclusions@#Myopic shift in PE was observed after combined phacovitrectomy for epiretinal membrane. ERM etiology, preoperative anterior chamber depth, and decrease of CMT were significantly associated with PE at postoperative 6 months. There was no difference in PE after surgery between the two groups defined by CMT (≥500 and <500 μm).

Comparison of Optical Coherence Tomography Biomarkers between Bevacizumab Good Responders and Nonresponders Who were Switched to Dexamethasone Implant in Diabetic Macular Edema

Purpose@#To compare volumetric optical coherence tomography (OCT) biomarkers in bevacizumab responsive and bevacizumab refractory diabetic macular edema (DME) patients switched to the dexamethasone implant to ultimately identify possible prognostic indicators. @*Methods@#Retrospective analysis of DME patients treated with bevacizumab were done. Patients were divided into those who showed response to bevacizumab (bevacizumab only group) and others who were switched to the dexamethasone implant due to lack of response to bevacizumab (switching group). Volumetric OCT biomarkers such as central macular thickness (CMT), inner and outer cystoid macular edema (CME) volume, serous retinal detachment (SRD) volume, retinal volume (CME + SRD volume) within the 6-mm Early Treatment of Diabetic Retinopathy Study circle were calculated. OCT biomarkers were followed up throughout treatment. @*Results@#Among total of 144 eyes, 113 patients were included in the bevacizumab only group and 31 patients were included in the switching group. Compared to the bevacizumab only group, the switching group showed higher baseline CMT (558.00 ± 209.60 µm vs. 454.96 ± 125.88 µm, p = 0.003), larger inner CME (6.02 ± 1.43 mm3 vs. 5.12 ± 0.87 mm3, p = 0.004) and SRD volume (0.32 ± 0.40 mm3 vs. 0.11 ± 0.09 mm3, p = 0.015) and higher proportion of patients with SRD (58.06% vs. 31.86%, p = 0.008). In the switching group, CMT, inner CME and SRD volume all showed significant reduction after switching to the dexamethasone implant. @*Conclusions@#DME with large SRD and inner nuclear layer edema volume may be more effectively treated with the dexamethasone implant than bevacizumab.

Endotracheal intubation in rabbits using a video laryngoscope with a modified blade.

Rabbits are being increasingly used as companion animals, and in research; thus, the need for proper veterinary care for rabbits has increased. Surgical access is more challenging in rabbits under inhalation anesthesia compared to other animals, such as dogs and cats. Rabbits have a very narrow and deep oral cavity, large incisors, and a large tongue. Moreover, their temporomandibular joint has limited mobility, making it more difficult to approach the larynx. Various methods have been proposed to overcome this difficulty. The video laryngoscope was introduced in 1999 and is useful when airway intubation is unsuccessful using a conventional laryngoscope. We postulated that a video laryngoscope with a modified size 1 Macintosh blade (McGrath MAC Video Laryngoscope, Medtronic, USA) would facilitate the intubation of New Zealand White rabbits. Sixteen specific-pathogen-free male New Zealand White rabbits weighing 3.45-4.70 kg were studied. All rabbits were intubated using the video laryngoscope. Typically, a 3.0 mm endotracheal tube was used for rabbits weighing < 4 kg, while a 3.5 mm tube was used in those weighing > 4 kg. During surgery, anesthesia was well maintained, and there were no major abnormalities in the animals' conditions. No rabbit developed breathing difficulties or anorexia after recovering from anesthesia. We established an intubation method using a video laryngoscope with a modified blade and stylet in the supine (ventrodorsal) position and successfully applied it in 16 rabbits. It is useful for training novices and for treating rabbits in veterinary hospitals with few staff members and animal research facilities where there are insufficient human resources.

Temporary Haptic Externalization and Four-point Fixation of Intraocular Lens in Scleral Fixation to Enhance Stability

PURPOSE: To report the results of a new technique for intraocular lens scleral fixation of temporary haptic externalization and four-point fixation for enhanced stability. METHODS: Two 10–0 polypropylene strands were fixed at two points 2 mm apart on each haptic of a conventional three-piece intraocular lens, using our previously reported method of temporary haptic externalization after injector implantation. Postoperative refractive outcome and stability were evaluated. RESULTS: Although the intraocular lens was fixed at a total of four points, no intraoperative difficulties were observed during the process. Patients showed successful fixation of the intraocular lens postoperatively. The fixed intraocular lens showed good centering and minimal tilting. When compared to the two-point fixation technique, postoperative astigmatism was significantly smaller in the four-point fixation group (1.80 ± 0.84 vs. 1.00 ± 0.0.50, p = 0.033). Lens-induced astigmatism calculated from subtraction of corneal astigmatism from total astigmatism was also significantly smaller in the four-point fixation group (2.23 ± 0.98 vs.1.17 ± 0.0.70, p = 0.043). No postoperative complications were identified during a mean follow-up period of 14.8 months (range, 10 to 19 months). CONCLUSIONS: By fixing the intraocular lens at two points on each side of the haptics, this method minimizes the tilting of the intraocular lens and thus decreases postoperative lens-induced astigmatism. Also, the possibility of intraocular lens dislocation in the long term might be decreased by this two-point fixation technique.

A Novel Fundus Image Reading Tool for Efficient Generation of a Multi-dimensional Categorical Image Database for Machine Learning Algorithm Training

BACKGROUND: We described a novel multi-step retinal fundus image reading system for providing high-quality large data for machine learning algorithms, and assessed the grader variability in the large-scale dataset generated with this system. METHODS: A 5-step retinal fundus image reading tool was developed that rates image quality, presence of abnormality, findings with location information, diagnoses, and clinical significance. Each image was evaluated by 3 different graders. Agreements among graders for each decision were evaluated. RESULTS: The 234,242 readings of 79,458 images were collected from 55 licensed ophthalmologists during 6 months. The 34,364 images were graded as abnormal by at-least one rater. Of these, all three raters agreed in 46.6% in abnormality, while 69.9% of the images were rated as abnormal by two or more raters. Agreement rate of at-least two raters on a certain finding was 26.7%–65.2%, and complete agreement rate of all-three raters was 5.7%–43.3%. As for diagnoses, agreement of at-least two raters was 35.6%–65.6%, and complete agreement rate was 11.0%–40.0%. Agreement of findings and diagnoses were higher when restricted to images with prior complete agreement on abnormality. Retinal/glaucoma specialists showed higher agreements on findings and diagnoses of their corresponding subspecialties. CONCLUSION: This novel reading tool for retinal fundus images generated a large-scale dataset with high level of information, which can be utilized in future development of machine learning-based algorithms for automated identification of abnormal conditions and clinical decision supporting system. These results emphasize the importance of addressing grader variability in algorithm developments.

Acepromazine inhibits hERG potassium ion channels expressed in human embryonic kidney 293 cells.

The effects of acepromazine on human ether-à-go-go-related gene (hERG) potassium channels were investigated using whole-cell voltage-clamp technique in human embryonic kidney (HEK293) cells transfected with hERG. The hERG currents were recorded with or without acepromazine, and the steady-state and peak tail currents were analyzed for the evaluating the drug effects. Acepromazine inhibited the hERG currents in a concentration-dependent manner with an IC50 value of 1.5 µM and Hill coefficient of 1.1. Acepromazine blocked hERG currents in a voltage-dependent manner between -40 and +10 mV. Before and after application of acepromazine, the half activation potentials of hERG currents changed to hyperpolarizing direction. Acepromazine blocked both the steady-state hERG currents by depolarizing pulse and the peak tail currents by repolarizing pulse; however, the extent of blocking by acepromazine in the repolarizing pulse was more profound than that in the depolarizing pulse, indicating that acepromazine has a high affinity for the open state of the channels, with a relatively lower affinity for the closed state of hERG channels. A fast application of acepromazine during the tail currents inhibited the open state of hERG channels in a concentration-dependent. The steady-state inactivation of hERG currents shifted to the hyperpolarized direction by acepromazine. These results suggest that acepromazine inhibits the hERG channels probably by an open- and inactivated-channel blocking mechanism. Regarding to the fact that the hERG channels are the potential target of drug-induced long QT syndrome, our results suggest that acepromazine can possibly induce a cardiac arrhythmia through the inhibition of hERG channels.

Oral Administration of Cilostazol Increases Ocular Blood Flow in Patients with Diabetic Retinopathy

PURPOSE: To investigate the effect of cilostazol on ocular hemodynamics and to determine whether the administration of cilostazol increases the ocular blood flow in patients with diabetic retinopathy. METHODS: This prospective observational study investigated the effect of orally administered cilostazol on diabetic retinopathy. Before and after administration for 1 week, pulsatile ocular blood flow (POBF) and retrobulbar hemodynamics were measured using a POBF analyzer and transcranial Doppler imaging, respectively. Visual acuity, intraocular pressure, and blood pressure were also evaluated before and after treatment. RESULTS: Twenty-five eyes of 25 patients were included in this study. POBF increased significantly (16.8 ± 4.6 µL/sec vs. 19.6 ± 6.2 µL/sec, p < 0.001) after administration of cilostazol, while no significant change was identified in visual acuity, intraocular pressure, and blood pressure. Mean flow velocity in the ophthalmic artery as measured with transcranial Doppler imaging also increased significantly after medication (23.5 ± 5.6 cm/sec vs. 26.0 ± 6.9 cm/sec, p = 0.001). The change in POBF directly correlated with the change in mean flow velocity (r = 0.419, p = 0.007). CONCLUSIONS: Cilostazol was effective in increasing ocular blood flow in patients with diabetic retinopathy, possibly by modulating retrobulbar circulation.

Comparison of Choroidal Hyperpermeability Change after Photodynamic Therapy and Ranibizumab for Chronic Central Serous Chorioretinopathy

PURPOSE: To compare changes in choroidal hyperpermeability after half-energy photodynamic therapy (PDT) and intravitreal ranibizumab in the treatment of chronic central serous chorioretinopathy (CSC). METHODS: Post-hoc analysis was performed in a randomized, controlled trial comparing half-energy PDT versus intravitreal ranibizumab for chronic CSC; during the experiments, the other treatment was available for salvage treatment if the original was unsuccessful at 3 months. A commercially available image analysis program (Adobe(R) Photoshop(R) CS6 [Adobe Systems, Inc., San Jose, CA]) was used for quantification of change in choriodal hyperpermeability on indocyanine green angiography after half-energy PDT or three consecutive intravitreal injections of ranibizumab. Post-treatment images were subtracted from pre-treatment images after adjustments were made to create images depicting the change in choroidal hyperpermeability with treatment. Integrated gray scale values per area in this image were used for analysis of change in choroidal hyperpermeability. RESULTS: The calculated change in choroidal hyperpermeability was significantly greater in the half-energy PDT group (17.36 +/- 8.74) than in the ranibizumab group (6.78 +/- 5.03) (p < 0.001). All eyes in the half-energy PDT group showed complete resolution of subretinal fluid, and no significant difference in change of choroidal hyperpermeability was found in eyes that received half-energy PDT as primary or salvage treatment. In the ranibizumab-treated group, subretinal fluid resolution was accomplished in 5 eyes, and these eyes showed a significantly larger decrease in choroidal hyperpermeability when compared with eyes showing poor response (10.31 +/- 4.00 vs. 2.74 +/- 2.16, p = 0.005). In the successfully treated eyes with ranibizumab, there was no significant difference in choroidal hypopermeability change when compared to half-energy PDT (p = 0.124). CONCLUSIONS: Using our novel method of analysis of change in choroidal hyperpermeability following treatment for chronic CSC, greater change was found in eyes with good response, and the superior outcome of half-energy PDT over ranibizumab may be attributed to greater influence on choroidal hyperpermeability.

Chlorogenic Acid Supplementation Improves Multifocal Electroretinography in Patients with Retinitis Pigmentosa

To evaluate the effect of chlorogenic acid supplementation in patients with retinitis pigmentosa, we evaluated objective change in visual function with multifocal electroretinography, along with visual acuity, visual field, standard electroretinography, and contrast sensitivity. Eighteen patients diagnosed with retinitis pigmentosa were enrolled in this prospective, non-comparative, single-arm study. Multifocal electroretinography, best-corrected visual acuity in Early Treatment Diabetic Retinopathy Study letters, total point score on visual field examination with Humphrey Field Analyzer II, electroretinography, and contrast sensitivity were measured and repeated after 3 months supplementation with chlorogenic acid. The amplitude of ring 5 was significantly higher on multifocal electroretinography after 3 months of chlorogenic acid supplementation (7.2 +/- 9.5 vs 8.3 +/- 10.8 nV/deg2, mean +/- standard deviation, P = 0.022). There were no significant changes in the best-corrected visual acuity, total point score on Humphrey Field Analyzer, 30 Hz flicker amplitude on standard electroretinography, or contrast sensitivity. Chlorogenic acid may have a beneficial effect on the peripheral area at the margins of retinal degeneration, and should be considered as an anti-oxidant for the management of retinitis pigmentosa.

Anti-VEGF-refractory Exudative Age-related Macular Degeneration: Differential Response According to Features on Optical Coherence Tomography

PURPOSE: To describe optical coherence tomography (OCT) characteristics of neovascular age-related macular degeneration (AMD) patients refractory to intravitreal anti-vascular endothelial growth factor (VEGF) injections (ranibizumab, bevacizumab) and their responses to alternative anti-VEGF agents or photodynamic therapy (PDT). METHODS: A retrospective review of 267 neovascular AMD patients treated with intravitreal anti-VEGF injections. RESULTS: Twenty patients (7.5%) were refractory to anti-VEGF injections (stationary or increased retinal exudation despite three or more monthly injections). They were grouped into either the extensive intraretinal fluid group (IRF group, 9 patients) or the subretinal fluid only group (SRF group, 11 patients) according to OCT findings. In the IRF group, response rates to subsequent treatment were 0% (0 / 7) for bevacizumab, 50% (3 / 6) for ranibizumab and 50% (3 / 6) for PDT +/- anti-VEGF. Three out of four bevacizumab-refractory patients showed response to ranibizumab as a secondary treatment. In the SRF group, response rates were lower with 0% (0 / 7) for bevacizumab, 22.2% (2 / 9) for ranibizumab and 28.6% (2 / 7) for PDT +/- anti-VEGF. One out of four bevacizumab-refractory patients responded to ranibizumab. The visual outcome was worse in the IRF group (median 20 / 1,000) than in the SRF group (median 20 / 100). CONCLUSIONS: In anti-VEGF-refractory neovascular AMD, patients with extensive IRF refractory to bevacizumab can be responsive to ranibizumab while patients with SRF may be refractory to both, suggesting a different pathophysiology and intraocular pharmacokinetics.

Chlorogenic Acid Decreases Retinal Vascular Hyperpermeability in Diabetic Rat Model

To evaluate the effect of chlorogenic acid (CGA), a polyphenol abundant in coffee, on retinal vascular leakage in the rat model of diabetic retinopathy, Sprague-Dawley rats were divided into four groups: controls, streptozotocin-induced diabetic rats, and diabetic rats treated with 10 and 20 mg/kg chlorogenic acid intraperitoneally daily for 14 days, respectively. Blood-retinal barrier (BRB) breakdown was evaluated using FITC-dextran. Vascular endothelial growth factor (VEGF) distribution and expression level was evaluated with immunohistochemistry and Western blot analysis. Expression of tight junction proteins, occludin and claudin-5, and zonula occludens protein, ZO-1 was also evaluated with immunohistochemistry and Western blot analysis. BRB breakdown and increased vascular leakage was found in diabetic rats, with increased VEGF expression and down-regulation of occludin, claudin-5, and ZO-1. CGA treatment effectively preserved the expression of occludin, and decreased VEGF levels, leading to less BRB breakdown and less vascular leakage. CGA may have a preventive role in BRB breakdown in diabetic retinopathy by preserving tight junction protein levels and low VEGF levels.

Evaluation of Corneal Biomechanical Properties Following Penetrating Keratoplasty Using the Ocular Response Analyzer

PURPOSE: To evaluate corneal biomechanical properties in eyes that had previously undergone penetrating keratoplasty (PK) using the ocular response analyzer (ORA). METHODS: We recruited 26 patients who had received unilateral PK. Corneal hysteresis (CH), corneal resistance factor (CRF), Goldmann-correlated intraocular pressure (IOPg), and cornea-compensated intraocular pressure (IOPcc) were measured with the ORA and were compared to the measurements from the contralateral eyes that did not undergo PK. RESULTS: The CH was 8.95+/-2.59 mmHg in eyes that underwent PK and 9.78+/-1.45 mmHg in the contralateral eyes that did not undergo PK (p=0.077). The CRF was 10.26+/-2.64 mmHg in post-PK eyes and 9.75+/-1.45 mmHg in the contralateral eyes (p=0.509), and the CH-CRF was significantly smaller in post-PK eyes (-1.31+/-2.32 mmHg in post-PK eyes vs. 0.03+/-0.88 mmHg in fellow eyes, p=0.016). The IOPg and IOPcc were significantly higher in the PK group than they were in the control group. The IOPcc's were 20.81+/-7.81 mmHg and 16.27+/-2.49 mmHg in post-PK and control eyes, respectively (p=0.011); and the IOPg's were 19.22+/-7.34 mmHg and 15.07+/-3.03 mmHg in post-PK and control eyes, respectively (p=0.019). The IOPcc-g's were 1.59+/-2.81 mmHg and 1.21+/-1.30 mmHg in post-PK and control eyes, respectively (p=0.412), and the central corneal thickness (CCT)'s were 489.11+/-90.60 microm and 556.24+/-42.84 microm in post-PK and control eyes, respectively (p=0.068). CONCLUSIONS: Following PK, CH tended to decrease while CRF tended to increase, significantly decreasing CH-CRF. A significantly higher intraocular pressure and a thinner CCT following PK may have contributed to the observed changes in these corneal biomechanical parameters.

Eye-Preserving Therapy in Retinoblastoma: Prolonged Primary Chemotherapy Alone or Combined with Local Therapy

PURPOSE: To evaluate the efficacy of primary chemotherapy combined with local therapy in the treatment of retinoblastomas not treatable with a single therapeutic method. METHODS: We performed a retrospective chart review of 227 patients diagnosed with retinoblastoma. Sixty-five eyes in 52 patients had tumors not treatable with a single therapeutic method and received primary chemotherapy combined with local therapy as needed. RESULTS: Tumor control and eye salvage was achieved in 34 of the 65 eyes; the probability of ocular survival was 46.56% using the Kaplan-Meier method. Forty-three of the 65 eyes were group D or E tumors, in which tumor control and eye salvage was achieved in 16 eyes. Twenty eyes were treated with chemotherapy only, while 28 eyes received one additional modality of local therapy, and 17 eyes received two modalities of local therapy. Of the eyes treated with chemotherapy only, tumor control was achieved in 5 eyes. CONCLUSIONS: Primary chemotherapy combined with local therapy can be effective and safe in the treatment of retinoblastomas otherwise untreatable with other therapeutic methods, such as group D and E retinoblastomas. More vigorous treatment with more local therapeutic methods combined may yield even better results.

A Case of Mitochondrial Myopathy With Cardiac Involvement

Mitochondrial myopathy is a disease caused by structural, biochemical or genetic disturbance of the mitochondria and this affects many organs, and it may also involve the cardiac muscles. We experienced a case of myocardial involvement in a 21 years old male patient with mitochondrial myopathy.

Prolactinoma presenting as giant cystic pituitary mass / 대한내과학회지

No abstract available.

Exploring the Nature of Caring in Hwa-Byung: Transpersonal Caring-Healing Model / 간호학회지

Hwa-Byung(HB) has been categorized as a Korean culture-bound syndrome that is prevalent in married women of low socioeconomic status. PURPOSE: The Purpose of the study was to search for the essence of HB and a caring-healing process of HB. Then the research result discusses whether the Transpersonal Caring-Healing Model has been congruent with it. METHOD: Case examples resulted from in-depth telephone counseling over a period of time at the Women's Hot Line with a client who is a housewife with HB. The counseling content was analyzed through Giorgi's method of descriptive phenomenology. RESULT: The core meaning of the essence of HB was 'injustice'; and essential themes were 'lack of reciprocity', 'infidelity' 'suppressed aggression and powerlessness' and 'need for recognition'. The core meaning of the essence of the caring-healing process was 'caring-healing experience(maintaining a trust relationship)'; and essential themes were 'active listening', 'empathy' and 'forming a therapeutic relationship'. According to Watson, 'active listening', 'empathy' and 'forming a therapeutic relationship' were identified as intentionality, intersubjectivity and transpersonal. CONCLUSION: Transpersonal caring can release inner power and strength and help the person to gain a sense of inner harmony. Transpersonal caring is as important to healing as are conventional treatment approaches and even more powerful in the long run.

Differential inhibition of endothelial cell proliferation and migration by urokinase subdomains: amino-terminal fragment and kringle domain

The serine protease urokinase-type plasminogen activator (uPA) is implicated in pericellular proteolysis in a variety of physiological and pathological processes including angiogenesis and tumor metastasis. The kringle domain of uPA (UK1) has proven to be an anti-angiogenic molecule with unknown mechanism and amino terminal fragment of uPA (u-ATF) with additional growth factor-like domain can be used for blocking interaction of uPA and uPA receptor. Here, we compared anti-angiogenic activities of these two molecules in vitro and in vivo. The recombinant u-ATF from E. coli and refolded in vitro was found to bind to uPAR with high affinity, whereas E. coli-derived UK1 showed no binding by Biacore analysis. In contrast to UK1 having potent inhibitory effect, u-ATF exhibited low inhibitory effect on bovine capillary endothelial cell growth (ED(50)>320 nM). Furthermore, u-ATF inhibition of VEGF-induced migration of human umbilical vein endothelial cell was far less sensitive (IC(50)= 600 nM) than those observed with UK1, and angiogenesis inhibition was marginal in chorioallantoic membrane. These results suggest that kringle domain alone is sufficient for potent anti- angiogenic activity and additional growth factor-like domain diverts this molecule in undergoing different mechanism such as inhibition of uPA/uPAR interaction rather than undergoing distinct anti- angiogenic mechanism driven by kringle domain.