Effects of oral glucosamine hydrochloride and mucopolysaccharide protein in a rabbit model of osteoarthritis.

AIM: The aim was to study whether oral glucosamine hydrochloride (GlcN.HCl) or mucopolysaccharide protein (MucoP) has a structure-modifying effect on an anterior cruciate ligament transection (ACLT) rabbit model of osteoarthritis (OA). METHODS: OA was surgically induced in the right knees of rabbits by transection of the ACLT. The left knees served as a sham-operated control. The animals were divided into four groups (n = 6 each): negative control (phosphate buffered saline, orally), positive control (oral celecoxib 10 mg/kg body weight/day), GlcN.HCl (oral 100 mg/kg/day) and MucoP (oral 100 mg/kg/day). Experimental animals were sacrificed after 8 weeks of treatment and the distal femur was removed for macroscopic examination, histological assessment, and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay of the OA rabbits. RESULTS: On gross morphology, severe lesions were observed in articular cartilage in the negative control group. In the GlcN.HCl and MucoP treatment groups, fibrillations and cartilaginous lesions were significantly (P < 0.05) decreased compared to the negative control group. In particular, degenerative changes in cartilage and chondrocyte cellularity were significantly reduced (P < 0.05) in the positive control (celecoxib) group, GlcN.HCl treatment group and MucoP treatment group compared with the negative control group. TUNEL assay showed that apoptotic chondrocytes were significantly suppressed in the celecoxib group. Similar significant (P < 0.05) results were seen in the GlcN.HCl group and MucoP group but apoptosis of chondrocytes were high in the negative control group. CONCLUSION: These data suggest that the protective effects of GlcN.HCl and MucoP may play a useful role in the clinical treatment of OA.

Correcting Severe Varus Deformity Using Trial Components During Total Knee Arthroplasty.

BACKGROUND: Extensive medial soft tissue release may be necessary to correct severe varus deformity during total knee arthroplasty (TKA). However, this procedure may result in instability. Here, we describe a novel soft tissue balancing technique, which can minimize medial release in severe varus deformity during TKA. METHODS: Fifty knees (40 patients) with hip-knee-ankle angle of more than 20° of varus were corrected using this technique (group 1). After achieving flexion gap balancing by needle puncturing and spreading of the superficial medial collateral ligament, extension gap balancing was obtained by gradual extension with the trial components in place. After group 1 was set, a one-to-one patient-matched control group who had mild varus deformity was selected by propensity score matching (50 knees, 48 patients, group 2). At postoperative 1 year, mediolateral laxity was compared between the 2 groups using the stress radiographs. Clinical outcomes were also compared using the Knee Society Score and Western Ontario and McMaster Universities Osteoarthritis Index score. RESULTS: There were no differences in mean medial and lateral laxities between groups 1 and 2 at 1 year after the operation (medial laxity: 2.3° ± 1.4° and 2.7° ± 1.3°, respectively, P = .310) (lateral laxity: 3.6° ± 1.7° and 3.2° ± 2.0°, respectively, P = .459). There were no significant differences in postoperative clinical scores and knee alignment. CONCLUSION: Our technique of obtaining extension gap balancing using trial components led to safe and effective balancing by avoiding unnecessary extensive release in severe varus deformity during TKA.

EXEL-7647 inhibits mutant forms of ErbB2 associated with lapatinib resistance and neoplastic transformation.

PURPOSE: Mutations associated with resistance to kinase inhibition are an important mechanism of intrinsic or acquired loss of clinical efficacy for kinase-targeted therapeutics. We report the prospective discovery of ErbB2 mutations that confer resistance to the small-molecule inhibitor lapatinib. EXPERIMENTAL DESIGN: We did in vitro screening using a randomly mutagenized ErbB2 expression library in Ba/F3 cells, which were dependent on ErbB2 activity for survival and growth. RESULTS: Lapatinib resistance screens identified mutations at 16 different ErbB2 amino acid residues, with 12 mutated amino acids mapping to the kinase domain. Mutations conferring the greatest lapatinib resistance cluster in the NH2-terminal kinase lobe and hinge region. Structural computer modeling studies suggest that lapatinib resistance is caused by multiple mechanisms; including direct steric interference and restriction of conformational flexibility (the inactive state required for lapatinib binding is energetically unfavorable). ErbB2 T798I imparts the strongest lapatinib resistance effect and is analogous to the epidermal growth factor receptor T790M, ABL T315I, and cKIT T670I gatekeeper mutations that are associated with clinical drug resistance. ErbB2 mutants associated with lapatinib resistance transformed NIH-3T3 cells, including L755S and T733I mutations known to occur in human breast and gastric carcinomas, supporting a direct mechanism for lapatinib resistance in ErbB2-driven human cancers. The epidermal growth factor receptor/ErbB2/vascular endothelial growth factor receptor inhibitor EXEL-7647 was found to inhibit almost all lapatinib resistance-associated mutations. Furthermore, no ErbB2 mutations were found to be associated with EXEL-7647 resistance and lapatinib sensitivity. CONCLUSIONS: Taken together, these data suggest potential target-based mechanisms of resistance to lapatinib and suggest that EXEL-7647 may be able to circumvent these effects.

Pharmacological abrogation of S-phase checkpoint enhances the anti-tumor activity of gemcitabine in vivo.

Chk1 and Chk2 kinases are critically involved in modulating DNA damage checkpoints. In particular, Chk1, a key activator of the S-phase DNA damage response, may be involved in resistance to genotoxic therapies that target DNA synthesis. We studied the in vitro and in vivo effects of EXEL-9844 (XL844), a potent, orally available, and specific inhibitor of Chk1 and Chk2, in combination with gemcitabine. In clonogenic assays using multiple cell lines in vitro, EXEL-9844 had only minor effects as a single agent but substantially enhanced gemcitabine-induced cell killing. Correspondingly, in PANC-1 cells, EXEL-9844 increased gemcitabine-induced H2AX phosphorylation, blocked Cdc25A phosphorylation, and induced premature mitotic entry. In a PANC-1 xenograft model, EXEL-9844 significantly enhanced gemcitabine antitumor activity but had limited effect as a single agent. Together, these data show that cell cycle checkpoint inhibitors may have significant clinical utility in potentiating the activity of gemcitabine.