Levels of glycosaminoglycans in the cerebrospinal fluid of healthy young adults, surrogate-normal children, and Hunter syndrome patients with and without cognitive impairment.

In mucopolysaccharidoses (MPS), glycosaminoglycans (GAG) accumulate in tissues. In MPS II, approximately two-thirds of patients are cognitively impaired. We investigated levels of GAG in cerebrospinal fluid (CSF) in different populations from four clinical studies (including NCT00920647 and NCT01449240). Data indicate that MPS II patients with cognitive impairment have elevated levels of CSF GAG, whereas those with the attenuated phenotype typically have levels falling between those of the cognitively affected patients and healthy controls

A phase I/II study of intrathecal idursulfase-IT in children with severe mucopolysaccharidosis II

Approximately two-thirds of patients with the lysosomal storage disease mucopolysaccharidosis II have progressive cognitive impairment. Intravenous (i.v.) enzyme replacement therapy does not affect cognitive impairment because recombinant iduronate-2-sulfatase (idursulfase) does not penetrate the blood-brain barrier at therapeutic concentrations. We examined the safety of idursulfase formulated for intrathecal administration (idursulfase-IT) via intrathecal drug delivery device (IDDD). A secondary endpoint was change in concentration of glycosaminoglycans in cerebrospinal fluid. Sixteen cognitively impaired males with mucopolysaccharidosis II who were previously treated with weekly i.v. idursulfase 0.5 mg/kg for ≥6 months were enrolled. Patients were randomized to no treatment or 10-mg, 30-mg, or 1-mg idursulfase-IT monthly for 6 months (four patients per group) while continuing i.v. idursulfase weekly. No serious adverse events related to idursulfase-IT were observed. Surgical revision/removal of the IDDD was required in 6 of 12 patients. Twelve total doses were administrated by lumbar puncture. Mean cerebrospinal fluid glycosaminoglycan concentration was reduced by approximately 90% in the 10-mg and 30-mg groups and approximately 80% in the 1-mg group after 6 months. These preliminary data support further development of investigational idursulfase-IT in MPS II patients with the severe phenotype who have progressed only to a mild-to-moderate level of cognitive impairment.Genet Med advance online publication 02 April 2015Genetics in Medicine (2015); doi:10.1038/gim.2015.36

A signal peptide peptidase is required for ER-symbiosome proximal association and protein secretion

Abstract During legume-rhizobia symbiosis, differentiation of the symbiosome (engulfed intracellular rhizobia) is necessary for successful nitrogen fixation. To control symbiosome differentiation, host cell subcellular components, e.g., ER (endoplasmic reticulum), must adapt robustly to ensure large-scale host protein secretion to the new organelle. However, the key components controlling the adaption of ER in nodule cells remain elusive. We report that Medicago BID1, a nodule-specific signal peptide peptidase (SPP), is central to ER structural dynamics and host protein secretion. In bid1, symbiosome differentiation is blocked. BID1 localizes specifically to the ER membrane and expresses exclusively in nodule cells with symbiosomes. In the wild type ER forms proximal association structures with symbiosomes, but not in bid1. Consequently, in bid1 excessive ER stress responses are induced and ER-to-symbiosome protein secretion is impaired. In summary, a nodule-specific SPP is necessary for ER-symbiosome proximal association, host protein secretion, and symbiosome differentiation

Morphological Characteristics of Postembryonic Early Development of the Thamnaconus septentrionalis

The morphological characteristics, ecological habits, and physiological structures of fish vary rapidly in the early growth and development stage. Understanding the changes in the postembryonic early development morphology of organs or tissues could provide basic information for the classification or identification of fish and the investigation or utilization of early resources. Morphological characteristic observation is one of the most commonly used methods. The existing studies on the early development of a certain trait in the late larval and juvenile stage are unclear. Most of the digital images are side views, resulting in a lack of dorsal views, ventral views, or local images, which is not conducive to the understanding of larval and juvenile. In this study, the variations in observable, measurable, and countable traits in postembryonic early development of Thamnaconus septentrionalis were explored using optical microscopy and stereomicroscopy. The observable traits included the developmental changes of yolk sac, scales, and notochord curvature. The measurable traits included the growth changes of total length, body length, body height, and oral fissure. The countable characters included the number of fins in the second dorsal fin, anal fin, caudal fin, and the number of pterygiophore. The results showed that at (23.0±1.0) ℃, the newly hatched larvae moved up and down in the water, with a phenomenon of convergent clustering under light irradiation; the pectoral fin appeared at 1 d, and the heart rate was approximately 50 beats/min; the optic capsule was completely black, the maxillar appeared, the oral fissure extended below the eyes, the yolk sac disappeared, and the first dorsal fin base appeared at 4 d; the girdle was single and colorless and began from the front edge of the belly, the oil globule disappeared, and the pelvic fin base appeared at 7 d; the first dorsal fin base and pelvic fin base broke membrane at 8 d; the first dorsal fin and pelvic fin were significantly longer than before, transparent rounded scales were observed at 9 d; swimming ability was improved, indicated by hovering back and forth gnawing pool wall, transversally conical barbs appeared at the base of the first dorsal fin at 11 d; the swim bladder and postcleithrum on both sides of the belly were clearly visible at 12 d; the second dorsal fin and anal fin pterygiophore appeared at 14 d, the second dorsal fin pterygiophore spans seven sarcomeres and the anal fin pterygiophore spans six sarcomeres. There are 10~12 melanin masses in the second dorsal fin and 5~6 in the anal fin, the conical teeth appeared on the upper and lower jaws at 16 d; the second dorsal fin and anal fin were differentiated at 19 d; the swim bladder fills about 25% of the abdominal cavity; notochord curves upward clearly and the stalk of caudal fin came out at 20 d; the fin strips of the second dorsal fin, anal fin, and caudal fin were well-developed at 24 d; the open mouth was round and changed from swallowing to sucking at 30 d, and the rainbow cells were colorful and the body surface near the gill was the most dense; the abdominal cavity has a strong metallic texture, the hard spine of the pelvic fin was degraded, close to the girdle at 39 d; 4~5 black bands were arranged regularly on the body, and the lateral line was consistent with the vertebral column at 50 d. The R2 values of the curve equations of measurable traits were all greater than 0.90. The height of the first dorsal fin increased exponentially 4~30 d. The visual sac increased linearly from 0~11 d to 11~20 d, and the increment of 0~11 d was less than that of 11~20 d, while the increment of 21~30 d was exponential. The oral fissure in 4~13 d and 22~30 d increased linearly, while that in 13~22 d decreased linearly. The total length, body length, and body height increased linearly before the inflection point and exponentially after the inflection point, which was concentrated in 19~23 d. The number of fin pterygiophore of the second dorsal fin was 31~36, while that in the anal fin was 33~35. The number of pectoral fins and caudal fins were 15~18 and 12, respectively. In conclusion, metamorphosis occurs in the postembryonic early development of T. septentrionalis, showing an allometric growth pattern, with a slow linear growth in the rotifer-feeding stage and a fast exponential growth in the artemia-feeding stage. The changes in the first dorsal fin and pelvic fin were correlated with age in days. The changes in the second dorsal and anal fins were correlated with the total length. This study enriches the early biological theory of T. septentrionalis and provides basic data for the investigation or utilization of its seedling breeding and resources. The partial enlarged detail is helpful for scholars to further understand the larval or juvenile fish and perform deep research

Whole Body and CNS Biodistribution of rhHNS in Cynomolgus Monkeys after Intrathecal Lumbar Administration: Treatment Implications for Patients with MPS IIIA

Mucopolysaccharidosis III type A (MPS IIIA; Sanfilippo syndrome), a genetic lysosomal disorder causing a deficiency of heparan N-sulfatase (HNS), leads to progressive cognitive decline from an early age. An effective enzyme replacement therapy (ERT) for MPS IIIA requires central nervous system (CNS) biodistribution. Recombinant human heparan N-sulfatase (rhHNS), an investigatory ERT for MPS IIIA, has been formulated for intrathecal (IT) administration since intravenous (IV) administration cannot cross the blood brain barrier (BBB) in sufficient amounts to have a therapeutic effect. In this study, systemic and CNS distribution of rhHNS in cynomolgus monkeys following IV and IT administration was evaluated by quantitation of rhHNS in serum, cerebral spinal fluid (CSF) and various tissues, and positron emission tomography (PET) imaging of live animals. Following IV administration, rhHNS levels were low to non-detectable in the CSF, and systemic clearance was rapid (≤2 h). With IT administration, rhHNS was observable in CNS tissues in ≤1 h, with varying Tmax (1–24 h). Appreciable systemic distribution was observed up to 7 days. This provides evidence that in this animal model, intrathecal administration of rhHNS delivers the replacement enzyme to therapeutically relevant tissues for the treatment of Sanfilippo Syndrome type A. Penetration into grey matter and cortex was 3–4 times greater than concentrations in white matter and deeper parenchymal regions, suggesting some limitations of this ERT strategy

Subcritical water extraction of betulinic acid from birch bark

An alternative and green method for extraction of betulinic acid (BA) from birch bark is here proposed as Subcritical Water Extraction (SWE). Using response surface methodology (RSM), optimization was conducted on reaction parameters for maximum extraction yield of BA. Optimal modifying conditions are as follows: extraction time of 27.37 min, extraction temperature at 184.52 degrees C and a solvent/solid ratio of 59.60 mL/g. Under optimal conditions, the maximum BA yield was 28.03 mg/10 g birch bark. Extracts by different methods (leaching extraction, reflux extraction, ultrasonic extraction, and SWE) were comparatively and quantitatively analyzed, using High Performance Liquid Chromatography (HPLC). Results show BA yields by SWE are higher than those obtained using ethanol, a traditional extraction method, plus the extract contained fewer impurities. Furthermore, the extraction processes for all four of the above methods were modeled and reviewed individually. (C) 2015 Elsevier B.V. All rights reserved