Age-related changes in the electrophoresis pattern of serum proteins in Korean indigenous calves.

This study aimed to investigate changes in the serum protein profiles of healthy newborn Korean calves and provide a reference index for these changes during growth. The serum protein composition of 15 healthy bovine calves aged 1 week to 4 months was analyzed using electrophoresis. The albumin to globulin (A/G) ratio increased until calves reached 2 months (1.3 ± 0.2) and then decreased until they reached 4 months (1.1 ± 0.1). Albumin concentrations increased up to month 2 (3.4 ± 0.1) and decreased until month 4 (3.2 ± 0.2). α-1 globulin concentrations decreased up to week 4 (0.4 ± 0.1) and then increased until month 2 (0.6 ± 0.1). γ globulin decreased until month 2 (0.7 ± 0.2) and then increased until month 4 (1.1 ± 0.3). These age-related changes in protein concentrations indicate the importance of considering age in calf blood tests. The γ globulin concentration was at its highest in the first week, due to colostrum intake, and decreased to reach its lowest at 2 months, when passive maternal immunity is also at its lowest. Therefore, provision of sufficient passive immunity through the colostrum in neonatal calves might enable healthy growth.

Cette étude visait à examiner les changements dans les profils de protéines sériques de veaux coréens nouveau-nés et de fournir un index de référence pour ces changements durant la croissance. La composition des protéines sériques de 15 veaux en santé âgés de 1 semaine à 4 mois a été analysée par électrophorèse. Le ratio albumine:globuline (A/G) a augmenté jusqu'à ce que les veaux atteignent l'âge de 2 mois (1,3 ± 0,2) et diminuait par la suite jusqu'à ce qu'ils aient 4 mois (1,1 ± 0,1). Les concentrations d'albumine augmentèrent jusqu'au mois 2 (3,4 ± 0,1) et diminuèrent ensuite jusqu'au mois 4 (3,2 ± 0,2). Les concentrations de α-1 globuline ont diminué jusqu'à la semaine 4 (0,4 ± 0,1) et ont par la suite augmenté jusqu'au mois 2 (0,6 ± 0,1). Les concentrations de γ globuline ont diminué jusqu'au mois 2 (0,7 ± 0,2) et ont par la suite augmenté jusqu'au mois 4 (1,1 ± 0,3). Ces changements dans les concentrations de protéines en fonction de l'âge démontrent l'importance de considérer l'âge lors des tests sanguins chez les veaux. La concentration de γ globuline était à son maximum durant la première semaine, due à l'ingestion de colostrum, et diminua par la suite pour atteindre un minimum à 2 mois, alors que l'immunité maternelle passive est à son plus bas. Ainsi, le fait de fournir suffisamment d'immunité passive via le colostrum chez les veaux nouveau-nés peut permettre une saine croissance.(Traduit par Docteur Serge Messier).

A novel apoptosis probe, cyclic ApoPep-1, for in vivo imaging with multimodal applications in chronic inflammatory arthritis.

Apoptosis plays an essential role in the pathophysiologic processes of rheumatoid arthritis. A molecular probe that allows spatiotemporal observation of apoptosis in vitro, in vivo, and ex vivo concomitantly would be useful to monitoring or predicting pathophysiologic stages. In this study we investigated whether cyclic apoptosis-targeting peptide-1 (CApoPep-1) can be used as an apoptosis imaging probe in inflammatory arthritis. We tested the utility of CApoPep-1 for detecting apoptotic immune cells in vitro and ex vivo using flow cytometry and immunofluorescence. The feasibility of visualizing and quantifying apoptosis using this probe was evaluated in a murine collagen-induced arthritis (CIA) model, especially after treatment. CApoPep-1 peptide may successfully replace Annexin V for in vitro and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay for ex vivo in the measurement of apoptotic cells, thus function as a sensitive probe enough to be used clinically. In vivo imaging in CIA mice revealed that CApoPep-1 had 42.9 times higher fluorescence intensity than Annexin V for apoptosis quantification. Furthermore, it may be used as an imaging probe for early detection of apoptotic response in situ after treatment. The CApoPep-1 signal was mostly co-localized with the TUNEL signal (69.6% of TUNEL+ cells) in defined cell populations in joint tissues of CIA mice. These results demonstrate that CApoPep-1 is sufficiently sensitive to be used as an apoptosis imaging probe for multipurpose applications which could detect the same target across in vitro, in vivo, to ex vivo in inflammatory arthritis.

Changes in serum protein electrophoresis profiles and acute phase proteins in calves with diarrhea.

Calf diarrhea leads to substantial economic losses in the livestock industry worldwide due to medical treatment costs, retarded growth performance, and even death. The objective of this study was to investigate changes in serum protein profiles and acute phase proteins in calves with diarrhea and identify the association between these changes and diarrhea. A total of 185 Korean beef calves were used and divided into 3 groups by age: 1 to 10 days (n = 46), 11 to 20 days (n = 65), and 21 to 30 days (n = 74). Blood and fecal samples were collected from each calf. Serum concentrations of total protein, protein fractions (albumin, α1-globulin, α2-globulin, ß-globulin, and γ-globulin), haptoglobin (Hp), and serum amyloid A (SAA) were analyzed. Compared to calves without diarrhea, calves with diarrhea had significantly lower albumin concentrations at 11 to 20 days and 21 to 30 days of age (P = 0.017 and P = 0.000, respectively) and significantly higher α1-globulin fractions at 21 to 30 days of age (P = 0.01). Interestingly, α2-globulin fractions were significantly higher in diarrheic calves in all age groups, whereas γ-globulin fractions were significantly lower in calves with diarrhea aged 1 to 10 days, compared with normal animals. In calves with diarrhea, the concentration of Hp was significantly higher, whereas SAA levels were not different between normal and diarrheic calves. In addition, a positive correlation was found between α2-globulin and Hp (P = 0.0004). Taken together, these results provide useful information about the use of serum protein profiles and Hp as prognostic and diagnostic markers for animal health status.

La diarrhée des veaux entraîne des pertes économiques substantielles dans l'industrie de l'élevage dans le monde entier en raison des coûts des traitements médicaux, du retard de croissance et même de la mort. L'objectif de cette étude était d'étudier les changements dans les profils des protéines sériques et les protéines de la phase aiguë chez les veaux souffrant de diarrhée et d'identifier l'association entre ces changements et la diarrhée. Un total de 185 veaux de boucherie coréens ont été utilisés et répartis en trois groupes par âge : 1 à 10 jours (n = 46), 11 à 20 jours (n = 65) et 21 à 30 jours (n = 74). Des échantillons de sang et de matières fécales ont été prélevés sur chaque veau. Les concentrations sériques de protéines totales, les fractions protéiques (albumine, α1-globuline, α2-globuline, ß-globuline et γ-globuline), d'haptoglobine (Hp) et d'amyloïde sérique A (SAA) ont été analysées. Par rapport aux veaux sans diarrhée, les veaux souffrant de diarrhée avaient des concentrations d'albumine significativement plus faibles à 11 à 20 jours et 21 à 30 jours d'âge (P = 0,017 et P = 0,000, respectivement) et des fractions d'α1-globuline significativement plus élevées à 21 à 30 jours d'âge (P = 0,01). Il est intéressant de noter que les fractions d'α2-globuline étaient significativement plus élevées chez les veaux diarrhéiques de tous les groupes d'âge, tandis que les fractions de γ-globuline étaient significativement plus faibles chez les veaux souffrant de diarrhée âgés de 1 à 10 jours, par rapport aux animaux témoins. Chez les veaux souffrant de diarrhée, la concentration de Hp était significativement plus élevée, tandis que les niveaux de SAA n'étaient pas différents entre les veaux normaux et diarrhéiques. De plus, une corrélation positive a été trouvée entre l'α2-globuline et Hp (P = 0,0004). Pris ensemble, ces résultats fournissent des informations utiles sur l'utilisation des profils de protéines sériques et de Hp comme marqueurs pronostiques et diagnostiques de l'état de santé des animaux.(Traduit par Docteur Serge Messier).

Alterations in serum protein electrophoresis profiles during the acute phase response in dogs with acute pancreatitis.

The quantification of serum proteins is a useful tool for diagnosing and monitoring various diseases that involve changes in the concentrations of these proteins. As canine acute pancreatitis (AP) accompanies the systemic inflammatory response syndrome, serum proteins such as C-reactive protein (CRP) have been used as inflammatory markers for dogs with AP. The goal of this study was to investigate the overall profiles of serum proteins by serum protein electrophoresis (SPE) and to determine the concentration of acute phase proteins (APPs) in dogs with AP in order to better understand serum protein profiles as diagnostic markers in these dogs. Decreased levels of albumin and increased levels of alpha-2 globulin were observed in dogs with AP by SPE. Among APPs, elevated concentrations of CRP, serum amyloid A (SAA), and haptoglobin were detected. The concentration of SAA was positively correlated with that of CRP, which suggests that SAA could be a sensitive marker of inflammation in dogs with AP, similar to CRP.

La quantification des protéines sériques est un outil utile pour diagnostiquer et suivre différentes pathologies qui impliquent des changements dans les concentrations de ces protéines. Comme la pancréatite aiguë (AP) accompagne le syndrome de réponse inflammatoire systémique, les protéines sériques telles que la protéine C-réactive (CRP) ont été utilisées comme marqueurs d'inflammation chez les chiens avec AP. L'objectif de la présente étude était d'examiner les profils globaux des protéines sériques par électrophorèse des protéines sériques (SPE) et de déterminer les concentrations des protéines de phase aiguë (APPs) chez les chiens avec AP afin de mieux comprendre les profils de protéines sériques comme marqueurs diagnostiques chez ces chiens. Des niveaux diminués d'albumine et des niveaux augmentés de globuline alpha-2 furent observés par SPE chez des chiens avec AP. Parmi les APPs, des concentrations élevées de CRP, d'amyloïde sérique A (SAA), et d'haptoglobine furent détectées. La concentration de SAA était corrélée positivement avec celle de CRP, ce qui suggère que SAA pourrait être un marqueur sensible d'inflammation chez les chiens avec AP, de manière similaire à la CRP.(Traduit par Docteur Serge Messier).

Acute phase response in bovine coronavirus positive post-weaned calves with diarrhea.

Bovine coronavirus (BCoV) is associated with severe diarrhea in calves, winter dysentery in adult cattle, and respiratory diseases in cattle of all ages. This study aimed to investigate the relationship between white blood cell counts and haptoglobin (Hp) and serum amyloid A (SAA) levels in post-weaned calves with diarrhea caused by BCoV and those that recovered from diarrhea. Blood and fecal samples were collected twice from the same animals; 17 post-weaned calves with diarrhea (first) and 15 post-weaned calves that recovered from diarrhea (second). Real-time polymerase chain reaction revealed that all 17 fecal samples from post-weaned calves with diarrhea and one out of 15 from diarrhea-recovered calves were positive for BCoV and negative for Cryptosporidium spp., Escherichia coli K99, Salmonella spp., bovine rotavirus, and bovine viral diarrhea virus. No Eimeria oocysts were detected using the flotation method. In comparison with post-weaned calves with diarrhea, in diarrhea-recovered calves, the lymphocyte count was significantly higher (P = 0.018), and the monocyte count was significantly lower (P = 0.001); however, the number of monocytes was still high. Post-weaned calves with diarrhea had a significantly higher Hp concentration (P < 0.001) compared with diarrhea-recovered calves. The results indicated that increased Hp concentration and monocytosis but not SAA may be associated with diarrhea caused by BCoV. The present study suggests that the monitoring of Hp concentration and monocyte count is useful in the diagnosis of post-weaned calves with diarrhea caused by BCoV in this field.

Stepwise inhibition of T cell recruitment at post-capillary venules by orally active desulfated heparins in inflammatory arthritis.

Identification of the structure-function relationship of heparin, particularly between 2-O-, 6-O-, and N-sulfation and its anticoagulant or anti-inflammatory activities, is critical in order to evaluate the biological effects of heparin, especially in conjunction with modifications for oral formulation. In this study, we demonstrated that removal of 2-O, 6-O, or N-desulfation and their hydrophobic modifications have differential effects on the blocking of interactions between sLeX and P-and L-selectins, with highest inhibition by 6-O desulfation, which was consistent with their in vivo therapeutic efficacies on CIA mice. The 6-O desulfation of lower molecular weight heparin (LMWH) retained the ability of LMWH to interfere with T cell adhesion via selectin-sLeX interactions. Furthermore, 6DSHbD coated on the apical surface of inflamed endothelium directly blocked the adhesive interactions of circulating T cells, which was confirmed in vivo by suppressing T cell adhesion at post-capillary venular endothelium. Thus, in series with our previous study demonstrating inhibition of transendothelial migration, oral delivery of low anticoagulant LMWH to venular endothelium of inflamed joint tissues ameliorated arthritis by the stepwise inhibition of T cell recruitment and provides a rationale for the development of modified oral heparins as innovative agents for the treatment of chronic inflammatory arthritis.

Endogenous inspired biomineral-installed hyaluronan nanoparticles as pH-responsive carrier of methotrexate for rheumatoid arthritis.

Methotrexate (MTX), an anchor drug for rheumatoid arthritis (RA), has been suffered from refractoriness and high toxicity limiting effective dosage. To mitigate these challenges, the ability to selectively deliver MTX to arthritis tissue is a much sought-after modality for the treatment of RA. In this study, we prepared mineralized nanoparticles (MP-HANPs), composed of PEGylated hyaluronic acid (P-HA) as the hydrophilic shell, 5ß-cholanic acid as the hydrophobic core, and calcium phosphate (CaP) as the pH-responsive mineral. Owing to the presence of CaP as the diffusion barrier, mineralized HANPs revealed the pH-responsiveness of release kinetics of MTX across neutral to acidic conditions. HANPs were internalized via receptor-mediated endocytosis in macrophages which involved molecular redundancy among major hyaladherins, including CD44, stabilin-2, and RHAMM. Following endocytosis, MP-HANPs loaded with doxorubicin revealed pH-dependent demineralization followed by dramatic acceleration of drug release into the cytosol compared to other HANPs. Furthermore, an in vivo study showed a significantly high paw-to-liver ratio of fluorescent intensity after systemic administration of MP-HANP-Cy5.5, indicating improved biodistribution of nanoparticles into arthritic paws in collagen-induced arthritis mice. Treatment with MTX-loaded MP-HANPs ameliorated inflammatory arthritis with remarkable safety at high dose of MTX. We highlight the distinct advantages of combining key benefits of biomineralization and PEGylation with HA-based nanoparticles for arthritis-selective targeting, thus suggesting MP-HANPs as a promising carrier of MTX for treatment of RA.

Robust Therapeutic Efficacy of Matrix Metalloproteinase-2-Cleavable Fas-1-RGD Peptide Complex in Chronic Inflammatory Arthritis.

OBJECTIVE: Therapeutic agents that are transformable via introducing cleavable linkage by locally enriched MMP-2 within inflamed synovium would enhance therapeutic efficacy on chronic inflammatory arthritis. Transforming growth factor-ß-inducible gene-h3 (ßig-h3), which consists of four fas-1 domains and an Arg-Gly-Asp (RGD) motif, intensifies inflammatory processes by facilitating adhesion and migration of fibroblast-like synoviocyte in the pathogenesis of rheumatoid arthritis (RA). The aim of this study was to investigate whether a MMP-2-cleavable peptide complex consisting of a fas-1 domain and an RGD peptide blocks the interaction between ßig-h3 and resident cells and leads to the amelioration of inflammatory arthritis. METHODS: We designed ßig-h3-derivatives, including the fourth fas-1 domain truncated for H1 and H2 sequences of mouse (MFK00) and MMP-2-cleavable peptide complex (MFK902). MMP-2 selectivity was examined by treatment with a series of proteases. MFK902 efficacy was determined by the adhesion and migration assay with NIH3T3 cells in vitro and collagen-induced arthritis (CIA) model using male DBA/1J mice in vivo. The mice were treated intraperitoneally with MFK902 at different dosages. RESULTS: MFK902 was specifically cleaved by active MMP-2 in a concentration-dependent manner, and ßig-h3-mediated adhesion and migration were more effectively inhibited by MFK902, compared with RGD or MFK00 peptides. The arthritis activity of murine CIA, measured by clinical arthritis index and incidence of arthritic paws, was significantly ameliorated after treatment with all dosages of MFK902 (1, 10, and 30 mg/kg). MFK902 ameliorated histopathologic deterioration and reduced the expression of inflammatory mediators simultaneously with improvement of clinical features. In addition, a favorable safety profile of MFK902 was demonstrated in vivo. CONCLUSION: The present study revealed that MMP-2-cleavable peptide complex based on ßig-h3 structure is a potent and safe therapeutic agent for chronic inflammatory arthritis, thus providing reliable evidence for a MMP-2-cleavable mechanism as a tissue-targeted strategy for treatment of RA.

Anthocyanin Profile and Antioxidant Activity of Various Berries Cultivated in Korea.

This study examined the anthocyanin composition and antioxidant activity of various berries cultivated in Korea: blueberry, crowberry, Korean black raspberry, mulberry, and strawberry. The anthocyanins in berries were identified by high-performance liquid chromatography (HPLC) analysis, and each component was quantitatively analyzed. Furthermore, the antioxidant activity of berries was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation decolorization, oxygen radical absorbance capacity (ORAC), and ferric ion reducing antioxidant (FRAP) assays. The results revealed that the total content of anthocyanins in crowberry was 35.1 mg/g of extract, which was higher than that in the other four major berry species (1.9-27.7 mg/g of extract). Nineteen anthocyanins were identified in the various berries. The major anthocyanins of crowberry were cyanidin-3-galactoside and delphinidin-3-galactoside, and those from Korean black raspberry were cyanidin-3-rutinoside and cyanidin-3-sambubioside-5-rhamnoside. These two berries also had relatively strong antioxidant activity accompanied by high total polyphenol contents. Thus, consumption of crowberry and Korean black raspberry may be beneficial in reducing the risk of developing lifestyle-related chronic diseases because of their strong antioxidant activity.

Great toenail dystrophy: a single-center experience and review of the literature.

BACKGROUND: There have been a few reported cases of congenital great toenail dystrophy (GTND), described as a congenital malalignment of the great toenails. However, acquired GTDN is rare, and has not been documented extensively. This study aimed to describe the clinical features of 21 patients with acquired GTND. METHODS: Twenty-one patients with acquired GTND who visited Yeouido St. Mary's Hospital between June 2005 and August 2012 were retrospectively reviewed. RESULTS: The mean patient age was 43.1 years (range, 17 to 88 years), and the cohort predominantly comprised women (18/21). In our experience, all acquired GTND patients presented with yellow or yellow-brownish chromonychia, onychotrophy, and onycholysis. Conservative treatment with tape methods and grinding, as well as nail extraction, was provided and yielded little improvement in any case. CONCLUSION: This study provides initial data on the nail changes affecting the great toenail, such as yellowish chromonychia, onychomadesis, and onycholysis. These data may help physicians to distinguish various nail disorders, including onychomycosis, congenital malalignment of the great toenails, and yellow nail syndrome.

Anaplastic lymphoma kinase-positive anaplastic large cell lymphoma arising in a patient with hypersensitivity to mosquito bites.

Hypersensitivity to mosquito bites is defined as the appearance of intense skin reactive lesions and systemic symptoms subsequent to mosquito bites. Most cases of hypersensitivity to mosquito bites reported thus far have been associated with chronic Epstein-Barr virus infection or natural killer cell leukemia/lymphoma. In this study, we describe the case of an 18-year-old Korean boy who had hypersensitivity to mosquito bites associated with primary systemic anaplastic lymphoma kinase-positive anaplastic large cell lymphoma. After a mosquito bite, the patient developed a progressive cutaneous nodule on his left lower leg and regional lymphadenopathy in the left inguinal area. The histopathological and immunohistochemical findings suggested anaplastic lymphoma kinase-positive anaplastic large cell lymphoma. Positron emission tomography-computed tomography revealed increased fluorodeoxyglucose uptake in the left T4 vertebrae, left external iliac lymph nodes, left inguinal lymph nodes, and lateral subcutaneous region of the left lower leg. According to the clinical, histopathological, and immunohistochemical findings, as well as the imaging data, the patient was diagnosed with primary systemic anaplastic lymphoma kinase-positive anaplastic large cell lymphoma. Consequently, the patient received a total of 6 cycles of cyclophosphamide + doxorubicin + vincristine + prednisolone chemotherapy at 3-week intervals, after which the lesions regressed.

Intracellular delivery of desulfated heparin with bile acid conjugation alleviates T cell-mediated inflammatory arthritis via inhibition of RhoA-dependent transcellular diapedesis.

Heparin has a potential regulatory role in inflammatory diseases. However, the anticoagulant activity and poor oral bioavailability of heparin limit its use as an anti-inflammatory agent. Conjugation of bis-deoxycholic acid to 6-O-desulfated low molecular weight heparin (6DSHbD) was efficiently internalized by activated endothelial cells via a 2-step model, in which heparin attaches to adhesion molecules that facilitate accessibility of the bile acid conjugate to membrane transporters. The critical role of P-selectin during endothelial cell uptake of 6DSHbD by arthritic tissue was confirmed in p-selectin(-/-) arthritic mice. Intracellular 6DSHbD inhibited transcellular diapedesis of T cells through activated endothelial cells and impaired both the formation of ICAM-1-rich docking structures at the T cell contact surface and subsequent cytoskeletal rearrangement. Furthermore, 6DSHbD blocked activation of RhoA-GTPase and phosphorylation of ezrin/radixin/moesin induced by ICAM-1 cross-linking on activated endothelial cells, thereby impairing lymphocyte transcellular transmigration. After oral administration 6DSHbD was preferentially delivered to inflamed joint tissue, particularly in and around post-capillary venular endothelium and inhibited effector T cell homing to arthritic joints. Aggravation of collagen-induced arthritis conferred by the transfer of effector T cells was suppressed by oral 6DSHbD. Thus, intracellular heparin exerts anti-inflammatory effects through the inhibition of RhoA-dependent transendothelial recruitment of T cells and may have applications in the treatment of chronic inflammatory arthritis.

Characteristic study on the chemical components of Korean curved ginseng products.

Dried ginseng (DG) is in fact the representing ginseng product in the worldwide market. Although it is made in various packages depending on the processing method, size and age of DG, basic scientific data reporting the chemical components are limited. In this study, 4-year-old curved ginseng (CG), one of the domestic DG products, was selected for further investigation. Eighty-six samples of 30 and 50 piece-grade CG, which are the most widely distributed in the market, were collected for 5 yr. Their major components, such as moisture, total sugar, acidic polysaccharides, total phenolic compounds, and saponins, were analyzed to figure out the standard quality characteristics. The moisture content of all CG samples was less than 15%. The total water-soluble sugar contents were 22.9% to 47.8% and 23.2% to 49.5% in the 30 and 50 piece-grade CG, respectively. The acidic polysaccharide contents were 3.6% to 6.7% and 2.9% to 6.9% in the 30 and 50 piece-grade CG, respectively. The total phenolic compound content was 0.4% to 0.5% in CG, regardless of the piece-grade. The crude saponin content, which represents the active component of ginseng, was over 2% in all samples. In 30 piece-grade CG samples, the contents of major ginsenosides, Rb1, Rf, and Rg1, were 2.2 to 4.7 mg/g, 0.4 to 1.3 mg/g, and 1.6 to 4.0 mg/g, respectively. The ginsenoside contents in 50 piece-grade CG samples were 2.1 to 3.9 mg/g (Rb1), 0.5 to 1.2 mg/g (Rf), and 1.3 to 3.4 mg/g (Rg1). Overall, since there were relatively high standard deviation and coefficient of variation in all the chemical component contents that were assessed, we found some difficulties in showing the CG standard chemical component characteristics by average, standard deviation, and other statistical analysis factors.

Comparison of single-port, two-port and four-port laparoscopic surgery for cyst enucleation in benign ovarian cysts.

OBJECTIVE: To compare the perioperative outcomes of three laparoscopic approaches for performing ovarian cyst enucleation. METHODS: A total of 148 patients underwent laparoscopic cyst enucleation at the CHA Gangnam Medical Center between September 2010 and May 2011. We reviewed retrospectively the medical records including patient demographics, operative outcomes and complications. RESULTS: We assigned the 148 patients into three groups: single-port (group A: 40), 2-port (group B: 30) and 4-port (group C: 78). There were no statistically significant differences in patient characteristics. The operation times were 90.4 ± 43.6, 74.7 ± 22.0 and 63.8 ± 30.5 min, and the estimated blood loss was 179.3 ± 253.9, 73 ± 75.2 and 89.9 ± 106.7 ml, respectively. Mean operation time was longer (p < 0.001) and estimated blood loss was higher (p = 0.005) in group A than in the other groups. There was no statistical difference in perioperative complications among the three groups. In group A, additional port insertion rate was higher than in groups B and C (p < 0.001). CONCLUSION: Single-port surgery required longer operation time, had a higher estimated blood loss and used additional ports more frequently during the operation than the other groups. However, 2-port surgery had no significant differences from 4-port surgery in the surgical outcomes. Therefore, 2-port surgery can be an alternative surgical option for 4-port surgery in ovarian cyst enucleation.

A matrix metalloproteinase 1-cleavable composite peptide derived from transforming growth factor ß-inducible gene h3 potently inhibits collagen-induced arthritis.

OBJECTIVE: Transforming growth factor ß-inducible gene h3 (ßIG-H3), which is abundantly expressed in rheumatoid synovium, and matrix metalloproteinases (MMP) play important roles in the pathogenesis of rheumatoid arthritis (RA). The aim of this study was to determine the therapeutic efficacy of ßIG-H3-derived peptides using MMP-1-dependent target tissue delivery in chronic inflammatory arthritis. METHODS: Peptides developed from ßIG-H3 derivatives, including the second and fourth YH peptides, the fourth fas-1 domain, the fourth fas-1 domain truncated for H1 and H2 sequences (dhfas-1), and an MMP-1- cleavable composite peptide (MFK24), were cloned. We confirmed the specificity of MFK24 cleavage by immunoblot analysis after treatment with different proteases. RESULTS: The YH18 peptide in the fourth fas-1 domain of ßIG-H3 was weakly effective in suppressing arthritis severity in mice with collagen-induced arthritis (CIA). Treatment with higher-dose dhfas-1 (30 mg/kg) showed remarkable efficacy, whereas treatment with a lower dose (10 mg/kg) resulted in only partial improvement. MFK24, a composite peptide consisting of dhfas-1 and RGD peptide linked by MMP-1 substrate, was cleaved specifically by MMP-1. The adhesion and migration of NIH3T3 cells mediated by ßIG-H3 were inhibited by MFK24 at a low concentration. MFK24 suppressed the adhesion of NIH3T3 cells more efficiently compared with murine dhfas-1 (MFK00) or RGD, either alone or in combination. The therapeutic efficacy of MFK24 in mice with CIA was remarkably enhanced, with consistently reduced expression of inflammatory mediators within joint tissue. CONCLUSION: This proof-of-concept study showed that an MMP-cleavable composite peptide, based on ßIG-H3 derivatives, had markedly improved therapeutic efficacy in chronic inflammatory arthritis, implicating a new expandable strategy for enhancement of the efficacy of 2 different active molecules in RA.

Orally active desulfated low molecular weight heparin and deoxycholic acid conjugate, 6ODS-LHbD, suppresses neovascularization and bone destruction in arthritis.

The regulation of angiogenesis is an interesting area to consider for novel therapeutic approaches to rheumatoid arthritis (RA). Chemically modified heparins have been developed as possible candidates for angiogenesis inhibitor; however, they have a major clinical drawback in exhibiting poor oral bioavailability. Here, orally absorbable O-desulfated low molecular weight heparin (ODS-LMWH) derivatives were newly synthesized by conjugating 2-O- or 6-O-desulfated LMWH with deoxycholic acid (DOCA) or bisDOCA (a dimer of DOCA), and their physicochemical properties, antiangiogenic potency and pharmacokinetic profiles were assessed. After selecting the best candidate among those derivatives, its therapeutic efficacy on arthritis was investigated in a murine collagen antibody-induced arthritis (CAIA) model. ODS-LMWH derivatives significantly inhibited the capillary-like tube formation of human umbilical vein endothelial cells (HUVECs) and basic fibroblast growth factor (bFGF)-induced angiogenesis in the Matrigel plug assay. Among all the compounds, 6ODS-LHbD showed the highest oral bioavailability in rats (19.3%). In the CAIA mouse model, 6ODS-LHbD (10 mg/kg, p.o., S.I.D.) significantly inhibited neovascularization in the joint, the increase of hind-paw thickness, and the structural damage in the bone. Therefore, 6ODS-LHbD would be a promising candidate for an orally active drug for the treatment of RA.