Myofascial Structural Integration Therapy on Gross Motor Function and Gait of Young Children with Spastic Cerebral Palsy: A Randomized Controlled Trial.

Though the cause of motor abnormalities in cerebral palsy is injury to the brain, structural changes in muscle and fascia may add to stiffness and reduced function. This study examined whether myofascial structural integration therapy, a complementary treatment that manipulates muscle and fascia, would improve gross motor function and gait in children <4 years with cerebral palsy. Participants (N = 29) were enrolled in a randomized controlled trial (NCT01815814, https://goo.gl/TGxvwd) or Open Label Extension. The main outcome was the Gross Motor Function Measure-66 assessed at 3-month intervals. Gait (n = 8) was assessed using the GAITRite(®) electronic walkway. Parents completed a survey at study conclusion. Comparing Treatment (n = 15) and Waitlist-Control groups (n = 9), we found a significant main effect of time but no effect of group or time × group interaction. The pooled sample (n = 27) showed a main effect of time, but no significantly greater change after treatment than between other assessments. Foot length on the affected side increased significantly after treatment, likely indicating improvement in the children's ability to approach a heel strike. Parent surveys indicated satisfaction and improvements in the children's quality of movement. MSI did not increase the rate of motor skill development, but was associated with improvement in gait quality.

Gait changes following myofascial structural integration (Rolfing) observed in 2 children with cerebral palsy.

Children with spastic cerebral palsy experience difficulty with ambulation. Structural changes in muscle and fascia may play a role in abnormal gait. Myofascial structural integration (Rolfing) is a manual therapy that manipulates muscle and soft tissues to loosen fascia layers, reposition muscles, and facilitate alignment. This study aimed to document (1) gait characteristics of 2 children with cerebral palsy and (2) effects of myofascial structural integration on their gait. Children received 3 months of weekly therapy sessions by an experienced practitioner. Gait parameters were recorded at baseline and after treatment using an electronic walkway. Children with cerebral palsy demonstrated abnormal velocity and cadence, decreased step length and single support times, and increased double support time. After treatment, both children demonstrated improvement for 3 months in cadence and double support time. The objective gait analyses demonstrated temporary improvements after myofascial structural integration in children with spastic cerebral palsy.

Activity-dependent release of tissue plasminogen activator from the dendritic spines of hippocampal neurons revealed by live-cell imaging.

Tissue plasminogen activator (tPA) has been implicated in a variety of important cellular functions, including learning-related synaptic plasticity and potentiating N-methyl-D-aspartate (NMDA) receptor-dependent signaling. These findings suggest that tPA may localize to, and undergo activity-dependent secretion from, synapses; however, conclusive data supporting these hypotheses have remained elusive. To elucidate these issues, we studied the distribution, dynamics, and depolarization-induced secretion of tPA in hippocampal neurons, using fluorescent chimeras of tPA. We found that tPA resides in dense-core granules (DCGs) that traffic to postsynaptic dendritic spines and that can remain in spines for extended periods. We also found that depolarization induced by high potassium levels elicits a slow, partial exocytotic release of tPA from DCGs in spines that is dependent on extracellular Ca(+2) concentrations. This slow, partial release demonstrates that exocytosis occurs via a mechanism, such as fuse-pinch-linger, that allows partial release and reuse of DCG cargo and suggests a mechanism that hippocampal neurons may rely upon to avoid depleting tPA at active synapses. Our results also demonstrate release of tPA at a site that facilitates interaction with NMDA-type glutamate receptors, and they provide direct confirmation of fundamental hypotheses about tPA localization and release that bear on its neuromodulatory functions, for example, in learning and memory.

Mitochondrial DNA integrity is not dependent on DNA polymerase-beta activity.

Mutations in mitochondrial DNA (mtDNA) are involved in a variety of pathologies, including cancer and neurodegenerative diseases, as well as in aging. mtDNA mutations result predominantly from damage by reactive oxygen species (ROS) that is not repaired prior to replication. Repair of ROS-damaged bases occurs mainly via base excision repair (BER) in mitochondria and nuclei. In nuclear BER, the two penultimate steps are carried out by DNA polymerase-beta (Polbeta), which exhibits both 5'-deoxyribose-5-phosphate (5'-dRP) lyase and DNA polymerase activities. In mitochondria, DNA polymerase-gamma (Polgamma) is believed to be the sole polymerase and is therefore assumed to function in mitochondrial BER. However, a recent report suggested the presence of Polbeta or a "Polbeta-like" enzyme in bovine mitochondria. Consequently, in the present work, we tested the hypothesis that Polbeta is present and functions in mammalian mitochondria. Initially we identified two DNA polymerase activities, one corresponding to Polgamma and the other to Polbeta, in mitochondrial preparations obtained by differential centrifugation and discontinuous sucrose density gradient centrifugation. However, upon further fractionation in linear Percoll gradients, we were able to separate Polbeta from mitochondria and to show that intact mitochondria, identified by electron microscopy, lacked Polbeta activity. In a functional test for the presence of Polbeta function in mitochondria, we used a new assay for detection of random (i.e., non-clonal) mutations in single mtDNA molecules. We did not detect enhanced mutation frequency in mtDNA from Polbeta null cells. In contrast, mtDNA from cells harboring mutations in the Polgamma exonuclease domain that abolish proofreading displayed a >or=17-fold increase in mutation frequency. We conclude that Polbeta is not an essential component of the machinery that maintains mtDNA integrity.