Collagenase treatment decreases muscle stiffness in cerebral palsy: A preclinical ex vivo biomechanical analysis of hip adductor muscle fiber bundles.

AIM: To determine the dose-response relationship of collagenase Clostridium histolyticum (CCH) on collagen content and the change in muscle fiber bundle stiffness after ex vivo treatment of adductor longus biopsies with CCH in children with cerebral palsy (CP). METHOD: Biopsy samples of adductor longus from children with CP (classified in Gross Motor Function Classification System levels IV and V) were treated with 0 U/mL, 200 U/mL, 350 U/mL, or 500 U/mL CCH; percentage collagen reduction was measured to determine the dose-response. Peak and steady-state stresses were determined at 1%, 2.5%, 5%, and 7.5% strain increments; Young's modulus was calculated. RESULTS: Eleven patients were enrolled (nine males, two females, mean age at surgery 6 years 5 months; range: 2-16 years). A linear CCH dose-response relationship was determined. Peak and steady-state stress generation increased linearly at 5.9/2.3mN/mm2 , 12.4/5.3mN/mm2 , 22.2/9.7mN/mm2 , and 33.3/15.5mN/mm2 at each percentage strain increment respectively. After CCH treatment, peak and steady-state stress generation decreased to 3.2/1.2mN/mm2 , 6.5/2.9mN/mm2 , 12.2/5.7mN/mm2 , and 15.4/7.7mN/mm2 respectively (p < 0.004). Young's modulus decreased from 205 kPa to 100 kPa after CCH (p = 0.003). INTERPRETATION: This preclinical ex vivo study provides proof of concept for the use of collagenase to decrease muscle stiffness in individuals with CP.

DNA Methylation Analysis Reveals Distinct Patterns in Satellite Cell-Derived Myogenic Progenitor Cells of Subjects with Spastic Cerebral Palsy.

Spastic type cerebral palsy (CP) is a complex neuromuscular disorder that involves altered skeletal muscle microanatomy and growth, but little is known about the mechanisms contributing to muscle pathophysiology and dysfunction. Traditional genomic approaches have provided limited insight regarding disease onset and severity, but recent epigenomic studies indicate that DNA methylation patterns can be altered in CP. Here, we examined whether a diagnosis of spastic CP is associated with intrinsic DNA methylation differences in myoblasts and myotubes derived from muscle resident stem cell populations (satellite cells; SCs). Twelve subjects were enrolled (6 CP; 6 control) with informed consent/assent. Skeletal muscle biopsies were obtained during orthopedic surgeries, and SCs were isolated and cultured to establish patient-specific myoblast cell lines capable of proliferation and differentiation in culture. DNA methylation analyses indicated significant differences at 525 individual CpG sites in proliferating SC-derived myoblasts (MB) and 1774 CpG sites in differentiating SC-derived myotubes (MT). Of these, 79 CpG sites were common in both culture types. The distribution of differentially methylated 1 Mbp chromosomal segments indicated distinct regional hypo- and hyper-methylation patterns, and significant enrichment of differentially methylated sites on chromosomes 12, 13, 14, 15, 18, and 20. Average methylation load across 2000 bp regions flanking transcriptional start sites was significantly different in 3 genes in MBs, and 10 genes in MTs. SC derived MBs isolated from study participants with spastic CP exhibited fundamental differences in DNA methylation compared to controls at multiple levels of organization that may reveal new targets for studies of mechanisms contributing to muscle dysregulation in spastic CP.

Transcriptional analysis of muscle tissue and isolated satellite cells in spastic cerebral palsy.

AIM: To analyze transcriptomes from muscle tissue and cells to improve our understanding of differences in gene expression and molecular function in cerebral palsy (CP) muscle. METHOD: In this case-control study, eight participants with CP (five males, three females; mean [SD] age 14y 2mo [1y 8mo]) and 11 comparison individuals (eight males, three females; mean [SD] age 14y 0mo [2y 6mo]) were enrolled after informed consent/assent and skeletal muscle was obtained during surgery. RNA was extracted from tissue and from primary satellite cells grown to form myotubes in vitro. RNA sequencing data were analyzed using validated informatics pipelines. RESULTS: Analysis identified expression of 6308 genes in the tissue samples and 7459 in the cultured cells. Significant differential expression between CP and control was identified in 87 genes in the tissue and 90 genes in isolated satellite cell-derived myotube cultures. INTERPRETATION: Both tissue and cell analyses identified differential expression of genes associated with muscle development and multiple pathways of interest. What this paper adds Expression differences were found in muscle tissue and in isolated muscle cells. There was low variability in expression among cells isolated from different muscles. Expression differences suggest complex functional alterations in spastic cerebral palsy.

Experimental Investigation on 3D Graphene-CNT Hybrid Foams with Different Interactions.

Due to the exceptional properties of graphene, numerous possibilities for real applications in various fields have been provided. However, it is a challenge to fabricate bulk graphene materials with properties arising from the nature of individual graphene sheets, and which assemble into monolithic three-dimensional structures. If 3D structured graphene foam were made instead of 2D structured graphene, it is expected that it would be a facile fabrication, with relatively low cost with the possibility of scale-up, and would maintain the intrinsic properties of graphene. To solve the weaknesses of 2D structured graphene, this study aimed to fabricate a 3D graphene-carbon nanotubes (CNT) hybrid foam. In this study, CNT was used to reinforce the graphene foams. In addition, two different surfactants, known as sodium dodecylbenzene sulphonate (SDBS) and cetyltrimethylammonium bromide (CTAB), were applied to help CNT dispersion. The π⁻π interaction was induced by SDBS/CNT, while ionic interaction was derived from CTAB/CNT. To confirm the charge effect with different surfactants, SEM, Zeta-potential, FT-IR, Raman spectroscopy, and compression tests were performed. When using a cationic surfactant, CTAB, compressive modulus, and strength increased due to the formation of relatively strong ionic bonding.

Hierarchical Porous Chitosan Sponges as Robust and Recyclable Adsorbents for Anionic Dye Adsorption.

Biomass waste treatment and detrimental dye adsorption are two of the crucial environmental issues nowadays. In this study, we investigate to simultaneously resolve the aforementioned issues by synthesizing chitosan sponges as adsorbents toward rose bengal (RB) dye adsorption. Through a temperature-controlled freeze-casting process, robust and recyclable chitosan sponges are fabricated with hierarchical porosities resulted from the control of concentrations of chitosan solutions. Tested as the adsorbents for RB, to the best of our knowledge, the as-prepared chitosan sponge in this work reports the highest adsorption capacity of RB (601.5 mg/g) ever. The adsorption mechanism, isotherm, kinetics, and thermodynamics are comprehensively studied by employing statistical analysis. Importantly and desirably, the sponge type of chitosan adsorbents exceedingly facilitates the retrieving and elution of chitosan sponges for recyclable uses. Therefore, the chitosan sponge adsorbent is demonstrated to possess dramatically squeezable capability with durability for 10,000 cycles and recyclable adsorption for at least 10 cycles, which provides an efficient and economical way for both biomass treatment and water purification.

Tensile properties of millimeter-long multi-walled carbon nanotubes.

There have been a number of theoretical and experimental studies on tensile properties of carbon nanotubes (CNT), reporting the Young's modulus of the individual CNT up to 1 TPa. Although CNT shows the promise to be used as reinforcement in a high modulus/strength composite material, it exhibits quite disappointing in terms of modulus or strength. Along with recent advance in CNT growth technique, we will be able to directly measure tensile properties of millimeter-long MWCNTs. This study firstly tackles the direct measurement of the tensile properties of millimeter-long MWCNTs that can be used as reinforcement in a composite system. A carefully designed tensile testing technique for the MWCNTs is developed, which allows us to obtain more accurate and reliable measured values. The average tensile strength and Young's modulus of the CNTs investigated in this study are measured to be 0.85 GPa and 34.65 GPa, respectively. Also, this work statistically investigates the effect of the CNT dimensions including length, diameter and volume on the tensile properties. To the best of our knowledge, this is the very first report on the tensile properties of macroscopically long and continuous CNTs.

Development and validation of a scale to measure symptoms of chronic graft-versus-host disease.

Chronic GVHD (cGVHD) affects 30% to 90% of surviving allogeneic transplant recipients. Thus far, no quality-of-life instruments have been developed to measure the effect of this common complication of allogeneic transplantation on patients' functioning and well-being. Using a prospective cohort of 107 patients with active cGVHD who completed the symptom scale at enrollment and at intervals of 3 and 6 months, we developed a 30-item symptom scale with 7 subscales to capture the cGVHD-specific symptom burden. The symptom scale correlated highly with patients' self-assessed mild, moderate, and severe cGVHD manifestations in cross-sectional analysis. Reliability (Cronbach's alpha = 0.79-0.90), test-retest (r2 = 0.28-0.93), and convergent and discriminant validity compared to the Medical Outcomes Study Short Form 36 (SF-36) and Functional Assessment of Chronic Illness Therapy with BMT subscale (FACT-BMT) were assessed and found to be adequate. Longitudinal assessments showed that changes in overall health status correlated best with changes in quality of life as measured by the SF-36 and FACT-BMT. In contrast, changes in cGVHD severity were best detected by changes in the symptom scale. We recommend that either the SF-36 or the FACT-BMT be combined with a cGVHD-specific symptom scale to measure the impact of cGVHD on patients' quality of life and that this endpoint be included in clinical trials testing cGVHD interventions. The cGVHD symptom scale is a short, simple, and valid measure of cGVHD manifestations and can be used to follow complication-specific symptoms using patient self-administered questionnaires.