Gi-Young Park

Glucosamine Hydrochloride Specifically Inhibits COX-2 by Preventing COX-2 N-Glycosylation and by Increasing COX-2 Protein Turnover in a Proteasome-dependent Manner *

COX-2 and its products, including prostaglandin E2, are involved in many inflammatory processes. Glucosamine (GS) is an amino monosaccharide and has been widely used for alternative regimen of (osteo) arthritis. However, the mechanism of action of GS on COX-2 expression remains unclear. Here we describe a new action mechanism of glucosamine hydrochloride (GS-HCl) to tackle endogenous and agonistdriven COX-2 at protein level. GS-HCl (but not GS sulfate, N-acetyl GS, or galactosamine HCl) resulted in a shift in the molecular mass of COX-2 from 72–74 to 66–70 kDa and concomitant inhibition of prostaglandin E2 production in a concentration-dependent manner in interleukin (IL)-1β-treated A549 human lung epithelial cells. Remarkably, GS-HCl-mediated decrease in COX-2 molecular mass was associated with inhibition of COX-2 N-glycosylation during translation, as assessed by the effect of tunicamycin, the protein N-glycosylation inhibitor, or of cycloheximide, the translation inhibitor, on COX-2 modification. Specifically, the effect of low concentration of GS-HCl (1 mm) or of tunicamycin (0.1 μg/ml) to produce the aglycosylated COX-2 was rescued by the proteasomal inhibitor MG132 but not by the lysosomal or caspase inhibitors. However, the proteasomal inhibitors did not show an effect at 5 mm GS-HCl, which produced the aglycosylated or completely deglycosylated form of COX-2. Notably, GS-HCl (5 mm) also facilitated degradation of the higher molecular species of COX-2 in IL-1β-treated A549 cells that was retarded by MG132. GS-HCl (5 mm) was also able to decrease the molecular mass of endogenous and IL-1β- or tumor necrosis factor-α-driven COX-2 in different human cell lines, including Hep2 (bronchial) and H292 (laryngeal). However, GS-HCl did not affect COX-1 protein expression. These results demonstrate for the first time that GS-HCl inhibits COX-2 activity by preventing COX-2 co-translational N-glycosylation and by facilitating COX-2 protein turnover during translation in a proteasome-dependent manner.

Sonoelastographic Evaluation of Medial Gastrocnemius Muscles Intrinsic Stiffness After Rehabilitation Therapy With Botulinum Toxin A Injection in Spastic Cerebral Palsy

OBJECTIVE To investigate intrinsic stiffness changes using real-time sonoelastography (RTS) in the medial gastrocnemius muscle (GCM) after rehabilitation therapy with botulinum toxin type A (BTA) injection in spastic cerebral palsy (CP). DESIGN Prospective study using ultrasonography and RTS. SETTING An inpatient rehabilitation clinic. PARTICIPANTS Children (N=17) with spastic CP (mean age, 57±22y, age range, 26-110mo). INTERVENTION Rehabilitation therapy and intramuscular injection of BTA in both medial and lateral GCMs. MAIN OUTCOME MEASURES RTS was obtained on the medial GCM, and the elastic pattern of the medial GCM was graded from RTS 1 (purple to green: soft) to RTS 4 (red: stiff) on the basis of color-scaled RTS. RTS score, color histogram, Modified Ashworth Scale (MAS) score of the ankle plantar flexor muscles, and Gross Motor Function Measure (GMFM) score were obtained before intervention and 4 weeks after intervention. The correlations among RTS score, GMFM, and MAS score were determined. Intrarater reliability was also evaluated. RESULTS Before and at 4 weeks after intervention, the mean RTS score decreased from 3.4 to 1.5 (P<.05), median red pixel intensity decreased from 112.5 to 101.3 (P<.05), median blue pixel intensity increased from 82.6 to 90.4 (P<.05), mean MAS score of the ankle decreased from 2.7 to 1.3 (P<.05), and mean GMFM score increased from 54.55% to 62.32%. Significant correlations were observed between the RTS score and the MAS score. Intrarater reliability was high. CONCLUSIONS Our results suggest that more information about the change of spastic muscle in CP after rehabilitation treatment with BTA may be gained by estimating muscle stiffness using RTS combined with clinical scale measurements.

Platelet-rich plasma limits the nerve injury caused by 10% dextrose in the rabbit median nerve

Introduction: We evaluated the effect of platelet‐rich plasma (PRP) injection in a rabbit model of dextrose‐induced median nerve injury. Methods: New Zealand white rabbits (n = 15) were divided randomly into 3 groups. Three different regimens (group 1: 0.1 ml saline; group 2: 10% dextrose with PRP; group 3: 10% dextrose with saline) were injected within the carpal tunnel. Electrophysiological and histological findings were evaluated 12 weeks after the injection. Results: The mean median motor latency in group 3 was significantly longer than that in groups 1 and 2. The cross‐sectional area of the median nerve and subsynovial connective tissue thickness in group 3 were significantly larger than those in groups 1 and 2. Conclusions: PRP injection may be effective in controlling median nerve injury, as demonstrated by improvement in electrophysiological and histological findings 12 weeks after dextrose injection. Muscle Nerve 49: 56–60, 2014

Regeneration of Full-Thickness Rotator Cuff Tendon Tear After Ultrasound-Guided Injection With Umbilical Cord Blood-Derived Mesenchymal Stem Cells in a Rabbit Model

Rotator cuff tendon tear is one of the most common causes of chronic shoulder pain and disability. In this study, we investigated the therapeutic effects of ultrasound‐guided human umbilical cord blood (UCB)‐derived mesenchymal stem cell (MSC) injection to regenerate a full‐thickness subscapularis tendon tear in a rabbit model by evaluating the gross morphology and histology of the injected tendon and motion analysis of the rabbits activity. At 4 weeks after ultrasound‐guided UCB‐derived MSC injection, 7 of the 10 full‐thickness subscapularis tendon tears were only partial‐thickness tears, and 3 remained full‐thickness tendon tears. The tendon tear size and walking capacity at 4 weeks after UCB‐derived MSC injection under ultrasound guidance were significantly improved compared with the same parameters immediately after tendon tear. UCB‐derived MSC injection under ultrasound guidance without surgical repair or bioscaffold resulted in the partial healing of full‐thickness rotator cuff tendon tears in a rabbit model. Histology revealed that UCB‐derived MSCs induced regeneration of rotator cuff tendon tear and that the regenerated tissue was predominantly composed of type I collagens. In this study, ultrasound‐guided injection of human UCB‐derived MSCs contributed to regeneration of the full‐thickness rotator cuff tendon tear without surgical repair. The results demonstrate the effectiveness of local injection of MSCs into the rotator cuff tendon.

Liquid chromatography-tandem mass spectrometry determination of baclofen in various biological samples and application to a pharmacokinetic study.

Baclofen is a structural analogue of γ-aminobutyric acid (GABA) that has been used for the treatment of spasticity since 1977. This study describes a simple and sensitive LC/MS/MS assay for the quantification of baclofen in rat plasma, urine, as well as various tissue samples. The assay utilized a simple protein precipitation and achieved lower limit of quantification (LLOQ) of 0.25ng/mL for rat plasma and brain samples and 2ng/mL for rat urine, liver and kidney samples. The assay was validated to demonstrate the specificity, linearity, recovery, LLOQ, accuracy, precision, and stability by using matrix matched quality control samples. There is no endogenous or exogenous peaks interfering with the analytes and matrix effects were minimized by optimized separation condition. The assay was linear over a concentration range of 0.25-500ng/mL for rat plasma and brain tissue, and 2-5000ng/mL for rat urine, kidney and liver with correlation coefficients >0.999. The mean intra- and inter-day assay accuracies were 94.6-104.6 and 96.0-103.6%, respectively. The mean intra- and inter-day precisions were 5.71 and 5.70%, respectively. The developed assay was successfully applied to a pharmacokinetic study and examined urinary excretion and tissue distribution of baclofen in rats following intravenous and oral administration.

Structural changes in the acromioclavicular joint measured by ultrasonography during provocative tests.

The acromioclavicular joint (ACJ) is a common source of pain and disability. Several provocative tests are used to diagnose ACJ lesions. The aim of this study was to evaluate anatomic changes of the ACJ using dynamic ultrasonography during provocative tests. Eighty ACJs of 40 healthy volunteers were subjected to dynamic ultrasonographic examinations. Examinations were performed in a resting position and during passive external rotation, cross body adduction, and active compression testing. Deep joint space distance and capsule displacement were measured ultrasonographically. Mean deep joint space distances during cross body adduction (6.4 ± 1.6 mm) and active compression (6.6 ± 1.8 mm) testing were significantly smaller than during passive external rotation testing (7.7 ± 1.7 mm, P < 0.01) or rest (8.1 ± 1.8 mm, P < 0.01). Mean capsule displacements during active compression (3.5 ± 0.8 mm) and cross body adduction (3.4 ± 0.7 mm) testing were significantly greater than those during passive external rotation testing (2.9 ± 0.7 mm, P < 0.01) or rest (2.5 ± 0.7 mm, P < 0.01). Dynamic ultrasonography revealed that the cross body adduction and active compression tests applied more stress to the ACJ than the passive external rotation test, which suggests that these two tests are likely to be more useful provocative tests for the diagnosis of ACJ lesions. Furthermore, the dynamic properties of ultrasonography could be used to estimate the suitability and validity of provocative tests in other musculoskeletal diseases. Clin. Anat. 22:580–585, 2009.

Ultrasonographic measurement of the axillary recess thickness in an asymptomatic shoulder

Purpose The purpose of this study was to measure the axillary recess (AR) thickness in an asymptomatic shoulder by using ultrasonography (US) and to analyze the factors affecting it. Methods We recruited 141 patients (52 males; 89 females; age, 57.7±9.9 years) with unilateral shoulder pain and performed US on the unaffected shoulder. Two physiatrists measured the AR thickness of the unaffected shoulder independently. All patients were examined in an upright sitting position with 90° shoulder abduction. The ultrasonographic transducer was placed longitudinally on the mid-axillary line and along the long axis of the humeral shaft. The factors affecting the AR thickness values were analyzed, and intra-class correlation coefficients were used for assessing the reproducibility of each measurement. Results The intrarater reliability values for the two physiatrists were 0.98 and 0.96, respectively. The inter-rater reliability of the mean AR thickness measurements was 0.91. The mean AR thickness in all subjects, males, and females was 2.8±0.6 mm, 3.1±0.6 mm, and 2.6±0.5 mm (P<0.01), respectively. No difference between the left and the right sides (males, P=0.086; females, P=0.535) or between the dominant and the non-dominant sides (males, P=0.173; female, P=0.244) was found. The AR thickness correlated positively with the height (r=0.313, P<0.01) and the weight (r=0.319, P<0.01). However, it did not correlate with the body mass index (r=0.152, P=0.077) or the age (r=-0.056, P=0.515). Conclusion US measurements of the AR thickness in asymptomatic shoulders demonstrated excellent intrarater and inter-rater reliabilities. The AR thickness showed anatomical variation with sex, height, and weight.

Comparison of Treatment Effects Between Children With Spastic Cerebral Palsy Under and Over Five Years After Botulinum Toxin Type A Injection

Objective To evaluate whether age influences a change in the spasticity of the ankle plantar flexor after botulinum toxin type A (BTA) injection in children with spastic cerebral palsy (CP). Methods Sixteen children with spastic CP were enrolled in the study. Seven children (group 1) were under 5 years of age, and nine (group 2) were over 5 years of age. They all received BTA injection in the gastrocnemius muscle (GCM) under ultrasound guidance. Passive range of motion (PROM) of ankle dorsiflexion, Modified Ashworth Scale (MAS) of the ankle plantar flexor, Gross Motor Function Measure (GMFM) and median red pixel intensity (RPI) of the medial GCM on real-time sonoelastography were measured at baseline (pre-injection) and 1-, 3-, and 6-month post-injection. Results In both groups, the mean PROM, MAS, and RPI were significantly improved after injection until 6-month post-injection. The change of PROM of ankle dorsiflexion in group 1 was significantly greater than that in group 2, until 6-month post-injection. The change in the MAS and GMFM between baseline and 6-month post-injection in group 1 was greater than that in group 2. The changes in the median RPI between baseline and 3- and 6-month post-injections were greater in group 1 than in group 2. Conclusion Our pilot study demonstrated the different changes in spasticity of the ankle plantar flexor after BTA injection based on age. Therefore, age may be considered when establishing a treatment plan using BTA injection for children with spastic CP.

Salivagram after gland injection of botulinum neurotoxin A in patients with cerebral infarction and cerebral palsy.

Pulmonary aspiration in patients with brain lesions may result from dysfunction of theswallowing process or gastroesophageal reflux. Inadequate protection of the airway fromoral secretions such as saliva may also induce aspiration. Chronic pulmonary aspiration ofsaliva can lead to substantial respiratory morbidity, including unexplained lung disease orrecurrent pneumonia, and is an important issue in the rehabilitation unit. In particular,patientswithneurologicdiseasewhoareinalong-termbedriddenstatecanbeatgreaterriskfor development of chronic salivary aspiration and pulmonary complications [1,2].Incontrast to aspiration during the swallowing process, continuous secretion of saliva and itsaspiration are difficult to control, even through modification of food consistency orcessation of oral feeding.Useofaradionuclidesalivagramtodetectpassiveaspirationofsalivawasintroducedby Heyman [3] in 1989. After sublingual instillation of a small amount of radiolabeledcolloid, serial images are evaluated for the presence of radiotracer uptake in thetracheobronchial tree, which is suggestive of aspiration. This test may be useful fordetection of aspiration of saliva and for patients with limitations of oral feeding or poorcooperation. For these reasons, a radionuclide salivagram has been used in infants orchildren with neurologic diseases, for example, cerebral palsy, who have recurrentpulmonary infection [4-6]. A radionuclide salivagram can be performed under morephysiological conditions without the challenge of an oral bolus for swallowing and isless invasive than a videofluoroscopic swallow study or a fiberoptic-endoscopic evalu-ation of swallowing[4].TherehasbeenanincreaseintheuseofbotulinumneurotoxinA(BoNT-A)injectionintosalivary glands for the treatment of drooling in patients with neurologic disease, includingcerebral palsy [7], Parkinson disease [8], and amyotrophic lateral sclerosis [9,10]. Variousobjective or subjective methods have been used to evaluate treatment response to BoNT-Ainjection.However,mostofthemethodsfocusonreductionofsialorrheaordroolingitself.FewstudiesassesspulmonaryaspirationofsalivaaftersalivaryglandinjectionwithBoNT-A[11]. We hypothesized that a positive radionuclide salivagram could be strong evidence ofaspirationofsalivaandhelpfultoassesstheindicationformoreintensivetreatment,suchasglandinjectionwithbotulinumtoxinorsurgicalexcision.Inourexperiencewith2patients,saliva aspiration resolved after administration of BoNT-A into salivary glands, as demon-strated on a follow-up radionuclide salivagram.

The novel anti-adipogenic effect and mechanisms of action of SGI-1776, a Pim-specific inhibitor, in 3T3-L1 adipocytes

The proviral integration site for moloney murine leukemia virus (Pim) kinases, consisting of Pim-1, Pim-2 and Pim-3, belongs to a family of serine/threonine kinases that are involved in controlling cell growth and differentiation. Pim kinases are emerging as important mediators of adipocyte differentiation. SGI-1776, an inhibitor of Pim kinases, is widely used to assess the physiological roles of Pim kinases, particularly cell functions. In the present study, we examined the effects of SGI-1776 on adipogenesis. The anti‑adipogenic effect of SGI‑1776 was measured by Oil Red O staining and AdipoRed assays. The effect of SGI‑1776 on the growth of 3T3‑L1 adipocytes was determined by cell count analysis. The effects of SGI‑1776 on the protein and mRNA expression of adipogenesis-related proteins and adipokines in 3T3‑L1 adipocytes were also evaluated by western blot analysis and RT‑PCR, respectively. Notably, SGI-1776 markedly inhibited lipid accumulation during the differentiation of 3T3-L1 preadipocytes into adipocytes. On a mechanistic level, SGI-1776 inhibited not only the expression of CCAAT/enhancer-binding protein-α (C/EBP-α), peroxisome proliferator-activated receptor-γ (PPAR-γ) and fatty acid synthase (FAS), but also the phosphorylation of signal transducer and activator of transcription-3 (STAT-3). Moreover, SGI-1776 decreased the expression of adipokines, including the expression of leptin and regulated on activation, normal T cell expressed and secreted (RANTES) during adipocyte differentiation. These findings demonstrate that SGI-1776 inhibits adipogenesis by downregulating the expression and/or phosphorylation levels of C/EBP-α, PPAR-γ, FAS and STAT-3.