Evaluating Functional Outcomes of Botulinum Toxin Type A Injection Combined with Occupational Therapy in the Upper Limbs of Children with Cerebral Palsy: A 9-Month Follow-Up from the Perspectives of Both Child and Caregiver

<div><p>Objective</p><p>To assess the effectiveness of combining botulinum toxin type A (BoNT-A) with functional occupational therapy (OT) at 9-month follow-up in children with cerebral palsy (CP) with bilateral upper limb impairments from the perspectives of both child and caregiver.</p><p>Methods</p><p>Twelve children with CP and their caregivers were assessed across 5 time points over 9 months based on the ICF after BoNT-A injection and functional OT in this open-label study.</p><p>Results</p><p>Significant differences were found across the 5 time points (<i>p</i> < .05) for both grasp and visual-motor integration with small effects (effect sizes = 0.12–0.24) and the self-care capability and performance of social function (<i>p <</i> .05). However, based on the effect sizes (0.02–0.14), no significant effects were found at the 4 post-test time points. Small effects were found on the psychological domain (effect sizes = 0.25–0.37) and environmental domains (effect size = 0.27) at follow-ups.</p><p>Conclusion</p><p>Combining a BoNT-A injection with OT not only reduced the muscle tone and increased ROM but also improved the upper limb function and self-care capability in children with CP. More importantly, these effects persisted for up to 9 months. Functional OT extends the effectiveness of a BoNT-A injection.</p></div

The spasticity distribution of muscles for the botulinum toxin type A injection for each child.

<p>The spasticity distribution of muscles for the botulinum toxin type A injection for each child.</p

Characteristics of the participants (N = 12).

<p>Characteristics of the participants (N = 12).</p

The dosage of botulinum toxin type A (Botox, onabotulinumtoxinA) injection in each muscle for each child.

<p>The dosage of botulinum toxin type A (Botox, onabotulinumtoxinA) injection in each muscle for each child.</p

ROM and spasticity changes from baseline after botulinum toxin type A injection.

<p>ROM and spasticity changes from baseline after botulinum toxin type A injection.</p

KMUP-1 Suppresses RANKL-Induced Osteoclastogenesis and Prevents Ovariectomy-Induced Bone Loss: Roles of MAPKs, Akt, NF-κB and Calcium/Calcineurin/NFATc1 Pathways

<div><p>Background</p><p>KMUP-1 is a xanthine derivative with inhibitory activities on the phosphodiesterase (PDE) 3,4 and 5 isoenzymes to suppress the degradation of cyclic AMP and cyclic GMP. However, the effects of KMUP-1 on osteoclast differentiation are still unclear. In this study, we investigated whether KMUP-1 inhibits osteoclastogenesis induced by RANKL in RAW 264.7 cells and bone loss induced by ovariectomy in mice, and the underlying mechanisms.</p><p>Principal Findings</p><p><i>In vitro</i>, KMUP-1 inhibited RANKL-induced TRAP activity, the formation of multinucleated osteoclasts and resorption-pit formation. It also inhibited key mediators of osteoclastogenesis including IL-1β, IL-6, TNF-α and HMGB1. In addition, KMUP-1 inhibited RANKL-induced activation of signaling molecules (Akt, MAPKs, calcium and NF-κB), mRNA expression of osteoclastogensis-associated genes (TRAP, MMP-9, Fra-1, and cathepsin K) and transcription factors (c-Fos and NFATc1). Furthermore, most inhibitory effects of KMUP-1 on RANKL-mediated signal activations were reversed by a protein kinase A inhibitor (H89) and a protein kinase G inhibitor (KT5823). <i>In vivo</i>, KMUP-1 prevented loss of bone mineral content, preserved serum alkaline phosphate and reduced serum osteocalcin in ovariectomized mice.</p><p>Conclusions</p><p>KMUP-1 inhibits RANKL-induced osteoclastogenesis <i>in vitro</i> and protects against ovariectomy-induced bone loss <i>in vivo</i>. These effects are mediated, at least in part, by cAMP and cGMP pathways. Therefore, KMUP-1 may have a role in pharmacologic therapy of osteoporosis.</p></div

Effects of KMUP-1 on RANKL-induced pit formation in mature osteocalsts.

<p>(<b>A</b>) Mature osteoclasts were treated with RANKL (10 ng/ml) and KMUP-1 <b>(</b>1–10<b> </b>µM) for 48 h. Pit formation on the disc was observed by optical microscopy. (<b>B</b>) Pit areas were quantified using Image Pro Plus analyzer Version 4.6 (Media Cybernetics Inc., MD). Each value represents the mean ± S.E.M. of three independent experiments, with triplicate determinations in each experiment. *<i>P</i><0.05, **<i>P</i><0.01 compared with RANKL alone.</p

Effects of KMUP-1 on RANKL-induced activations of c-Fos and the calcium/calcineurin/NFATc1 pathway.

<p>(<b>A</b>) RAW264.7 cells were cultured for 24 h with RANKL (10 ng/ml) and KMUP-1 (0–10 µM). Cell lysates were then analyzed by Western blotting with antibodies against c-Fos, NFATc1, calcineurin and actin. (<b>B</b>–<b>D</b>) The expressions of these proteins were quantified by densitometry. (<b>E</b>) In RAW264.7 cells, RANKL did not induce Ca²⁺ oscillation. Each color indicates an individual cell in the same field. (<b>F, G)</b> In osteoclasts, KMUP-1 inhibited Ca²⁺ oscillation evoked by RANKL. Pretreatment with KMUP-1 (10 µM), similar to calcium chelator BAPTA (10 µM), significantly reduced the amplitude of oscillation induced by RANKL. Each value represents the mean ± S.E.M. of three independent experiments, with triplicate determinations in each experiment. ^##<i>P</i><0.01 compared with control; *<i>P</i><0.05, **<i>P</i><0.01 compared with RANKL alone.</p

Effects of KMUP-1 on RANKL-induced activations of MAPKs and Akt pathways.

<p>(<b>A</b>) Time course analysis of RANKL-induced phosphorylation of MAPKs and Akt showed that their activations were maximized at 15 min or 30 min, respectively. (<b>B</b>–<b>E</b>) RAW264.7 cells were pretreated with KMUP-1 for 24 h followed by stimulation with RANKL (10 ng/ml) for 15 min (MAPKs) or 30 min (Akt). The cell lysates were analyzed by Western blotting. Each value represents the mean ± S.E.M. of three independent experiments, with triplicate determinations in each experiment. ^##<i>P</i><0.01 compared with control; *<i>P</i><0.05, **<i>P</i><0.01 compared with RANKL alone.</p

Effects of KMUP-1 on changes in histomorphometric and biochemical markers of bone turnover in OVX mice.

<p>Histomorphometric data and serum biochemical markers were compared in sham-operated, OVX mice, and OVX+KMUP-1 mice. (<b>A</b>) Tb.N, trabecular number, (<b>B</b>) Tb.Th, trabecular thickness, (<b>C</b>) Tb.Sp, trabecular separation, (<b>D</b>) ALP, serum alkaline phosphatase and (<b>E</b>) osteocalcin. All values are expressed as mean ± S.E.M. ^#<i>P</i><0.05 compared with the Sham group;^*<i>P</i><0.05, ^**<i>P</i><0.01 compared with the OVX group.</p