Feasibility and safety of Robot Suit HAL treatment for adolescents and adults with cerebral palsy.

To investigate whether Robot Suit HAL treatment (HAL-T) is safe and feasible for gait disorders in adolescents and adults with cerebral palsy (CP). We tested HAL-T in adolescents and adults with bilateral spastic CP (four men, four women; mean age: 18.2 years). Three participants were classified as level III under the Gross Motor Function Classification System (GMFCS), and five were classified as level IV. The participants underwent HAL-T twice per week for 4 weeks. The outcome measures, which were assessed before and after HAL-T, included comfortable gait speed (CGS), step length (SL), cadence, and GMFCS level. Adverse events were noted. All participants completed the HAL-T sessions despite some mild adverse events occurring. The mean increases in CGS, SL, and cadence were 0.19 ±â€¯0.14 m/s (p = 0.006), 0.09 ±â€¯0.08 m (p = 0.020), and 18.0 ±â€¯15.9 steps/min (p = 0.015), respectively. HAL-T is safe and feasible for gait disorders in patients with CP. HAL-T can significantly improve CGS, SL, and cadence and may be effective for improving walking ability in adolescents and adults with CP.

Overexpression of a Gene Involved in Phytic Acid Biosynthesis Substantially Increases Phytic Acid and Total Phosphorus in Rice Seeds.

The manipulation of seed phosphorus is important for seedling growth and environmental P sustainability in agriculture. The mechanism of regulating P content in seed, however, is poorly understood. To study regulation of total P, we focused on phytic acid (inositol hexakisphosphate; InsP6) biosynthesis-related genes, as InsP6 is a major storage form of P in seeds. The rice (Oryza sativa L.) low phytic acid mutant lpa1-1 has been identified as a homolog of archael 2-phosphoglycerate kinase. The homolog might act as an inositol monophosphate kinase, which catalyzes a key step in InsP6 biosynthesis. Overexpression of the homolog in transgenic rice resulted in a significant increase in total P content in seed, due to increases in InsP6 and inorganic phosphates. On the other hand, overexpression of genes that catalyze the first and last steps of InsP6 biosynthesis could not increase total P levels. From the experiments using developing seeds, it is suggested that the activation of InsP6 biosynthesis in both very early and very late periods of seed development increases the influx of P from vegetative organs into seeds. This is the first report from a study attempting to elevate the P levels of seed through a transgenic approach.

Synthesis of zeolites from paper sludge ash and their ability to simultaneously remove NH4+ and PO43-.

The objective of this study was to investigate the synthesis of zeolites from paper sludge ash, and to evaluate their applications for water purification to simultaneously remove NH4+ and PO43- in aqueous solution. Paper sludge ash had a low abundance of Si and a significant Ca content, due to the presence of calcite that was used as paper filler. Na2SiO3 was added to the NaOH solution to increase Si content in order to synthesize zeolites with high cation exchange capacity. The original ash without addition of Si yielded hydroxysodalite with a cation exchange capacity of ca. 50 cmol/kg. Addition of Si to the solution yielded Na-P1 (zeolite-P) with a higher cation exchange capacity (ca. 120 cmol/kg). The observed concentrations of Si and Al in the solution during the reaction explain crystallizations of these two phases. The product with Na-P1 could simultaneously remove NH4+ and PO43- from the solution between pH 4 and 9.

Selective enhancer of tumor vascular permeability for optimization of cancer chemotherapy.

Clinical approach using tumor necrosis factor-alpha (TNF-alpha) as selective destruction against tumor endothelial cells and selective enhancer of tumor vascular permeability for effective accumulation of antitumor chemotherapeutic agents has attracted attention. However, the clinical application of TNF-alpha as a systemic antitumor agent has been limited because of toxic side-effects. To systemically use TNF-alpha as an antitumor agent and the selective enhancer of tumor vascular permeability, we assessed the usefulness of PEGylated TNF-alpha (PEG-TNF-alpha). PEG-TNF-alpha at a dose of 1000 JRU showed marked hemorrhagic necrosis in S-180 tumors without side-effects due to selective destruction of tumor vasculature, whereas wild-type TNF-alpha at a dose of 10,000 JRU showed a little hemorrhagic necrosis with severe side-effects. PEG-TNF-alpha induced the enhancement of tumor vascular permeability. The permeability was increased at 1 h, after an i.v. injection of PEG-TNF-alpha and returned to the basal level at 2 h. In addition, high molecular weight of PEG (molecular weight; 500K) accumulated in tumor tissue as well as low molecular weight of PEG (molecular weight; 12K). On the other hand, PEG-TNF-alpha didn't affect the permeability of normal tissue and inflammation site. This data suggested that PEG-TNF-alpha was useful agent as selective enhancer of tumor vascular permeability with safe.

Site-specific PEGylation of a lysine-deficient TNF-alpha with full bioactivity.

Addition of polyethylene glycol to protein (PEGylation) to improve stability and other characteristics is mostly nonspecific and may occur at all lysine residues, some of which may be within or near an active site. Resultant PEGylated proteins are heterogeneous and can show markedly lower bioactivity. We attempted to develop a strategy for site-specific mono-PEGylation using tumor necrosis factor-alpha (TNF-alpha). We prepared phage libraries expressing TNF-alpha mutants in which all the lysine residues were replaced with other amino acids. A fully bioactive lysine-deficient mutant TNF-alpha (mTNF-alpha-Lys(-)) was isolated by panning against TNF-alpha-neutralizing antibody despite reports that some lysine residues were essential for its bioactivity. mTNF-alpha-Lys(-) was site-specifically mono-PEGylated at its N terminus. This mono-PEGylated mTNF-alpha-Lys(-), with superior molecular uniformity, showed higher bioactivity in vitro and greater antitumor therapeutic potency than randomly mono-PEGylated wild-type TNF-alpha. These results suggest the usefulness of the phage display system for creating functional mutant proteins and of our site-specific PEGylation approach.