Glycosphingolipids (GSLs) from brain homogenates were extracted with chloroform:methanol (1:2, v/v) overnight at 4°C and further purified using solid-phase C18 columns (Telos, Kinesis, UK). GSLs were dried down under nitrogen and treated with either Cerezyme® (Genzyme, Cambridge, MA) to obtain glucose from GlcCer, or ceramide glycanase (prepared in house from the medicinal leech Hirudo medicinalis/verbena) to obtain oligosaccharides from other GSLs. Liberated glucose and free glycans were then fluorescently-labelled with anthranillic acid (2AA). Excess 2AA label was removed using DPA-6S SPE columns (Supelco, PA, USA). Purified 2AA-labelled glucose and 2AA-labelled glycans were separated and quantified by normal phase high-performance liquid chromatography (NP-HPLC) 34 (link). The NP-HPLC system consisted of a Waters Alliance 2695 separations module and an in-line Waters 2475 multi λ-fluorescence detector set at Ex λ360nm and Em λ425nm. The solid phase used was a 4.6 x 250mm TSK gel-Amide 80 column (Anachem, Luton, UK). Results were normalized to protein content determined using bicinchoninic acid (BCA) assay.
Cerezyme
Manufactured by Sanofi
Sourced in United States
Cerezyme is a laboratory product used for the treatment of Gaucher disease. It is an enzyme replacement therapy that provides the missing enzyme, glucocerebrosidase, to individuals with Gaucher disease. The core function of Cerezyme is to help manage the symptoms and complications associated with Gaucher disease.
Automatically generated - may contain errors
Lab products found in correlation
Alliance 2695 separations module, by Waters Corporation (3 mentions)
2475 multi λ fluorescence detector, by Waters Corporation (2 mentions)
Dpa 6s spe columns, by Merck Group (2 mentions)
Ambroxol, by Merck Group (1 mentions)
Isofagomine, by LGC (1 mentions)
4 methylumbelliferyl β d glucopyranoside, by Merck Group (1 mentions)
Triton x 100, by Nacalai Tesque (1 mentions)
F 4500 fluorescence spectrophotometer, by Hitachi (1 mentions)
8 protocols using cerezyme
1
Glycosphingolipid Quantification from Brain
2
Unilateral Putamen Delivery of Cerezyme in Parkinson's Disease
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This open label, prospective, proof-of-concept, phase I trial (clinicaltrials.gov identifier: NCT04370665) was designed to study the safety and feasibility of delivering beta-glucocerebrosidase (Cerezyme; Genzyme, Cambridge, MA, USA) to the putamen unilaterally in patients with PD. The treatment side was chosen based on the relative lateralized severity of the patient’s symptoms. The procedure was repeated once every two weeks for a total of 3 treatment sessions per patient. Bi-weekly scheduling was based on the recommended dosage on the drug label [42 ]. The study was approved by our institutional Research Ethics Board (project ID: 368–2018). All patients and their primary caregivers provided informed consent prior to study enrolment. Candidates were excluded from the trial if they were contraindicated to MRI or MR/ultrasound contrast agents, were at an increased risk of cerebral bleeding, or had active intracranial, cardiovascular, pulmonary, or renal disease. The inclusion and exclusion criteria, as well as primary outcomes, were reported previously [24 (link)]. This report focuses on technical details of FUS treatments.
3
Quantification of Glycosphingolipids in Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GlcCer and downstream GSLs were analyzed essentially as described by Neville and coworkers (62 (link)). Lumbar spinal cord and soleus muscle were homogenized in water using an Ultraturax T25 probe homogenizer (IKA, Germany). Lipids from tissue homogenates were extracted with chloroform and methanol. The GSLs were then further purified using solid-phase C18 columns (Telos, Kinesis, UK). After elution, the GSL fractions were split in half, dried down under nitrogen at 42°C and treated with either Cerezyme® (Genzyme, Cambridge, MA) to obtain glucose from GlcCer or ceramide glycanase (prepared in house from the medicinal leech Hirudo medicinalis/verbena) to obtain oligosaccharides from other GSLs. The liberated glucose and free glycans were then fluorescently-labeled with anthranillic acid (2-AA). To remove excess free label, samples were separated in 6S SPE columns (Supelco, PA, USA). Purified 2AA-labeled glucose and 2AA-labeled oligosaccharides were separated and qualified by normal-phase high-performance liquid chromatography (NP-HPLC) as previously described (62 (link)). The solid phase used was a 4.6 × 250 mm TSK gel-Amide 80 column (Anachem, Luton, UK). The HPLC system consisted of a Waters Alliance 2695 separations module and an in-line Waters 2475 multi λ_fluorescence detector set at Ex λ_360 nm and Em λ_425 nm.
4
Glycosphingolipid Quantification from Brain
5
Imiglucerase Supply Constraint Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This was an observational, retrospective, multicenter, national, non-comparative study.
All patients with GD1 or GD3, treated with imiglucerase (Cerezyme® [Ceredase® before 1996], Genzyme-a SANOFI company) for at least 6 months at the time of their therapeutic scheme modification, which was due to the imiglucerase supply constraint, could potentially be included in this study. To this aim, all French physicians treating at least one patient with GD at the time of the study implementation were proposed to participate as study investigators.
The patient (or legal guardian) was informed and agreed to participation. The study was approved by the French Healthcare Research Data Processing Advisory Board, the French National Data Protection Commission, and the Institutional Review Board of Paris North Hospitals. It was conducted in accordance with the French Good Epidemiological Practice guidelines.
All patients with GD1 or GD3, treated with imiglucerase (Cerezyme® [Ceredase® before 1996], Genzyme-a SANOFI company) for at least 6 months at the time of their therapeutic scheme modification, which was due to the imiglucerase supply constraint, could potentially be included in this study. To this aim, all French physicians treating at least one patient with GD at the time of the study implementation were proposed to participate as study investigators.
The patient (or legal guardian) was informed and agreed to participation. The study was approved by the French Healthcare Research Data Processing Advisory Board, the French National Data Protection Commission, and the Institutional Review Board of Paris North Hospitals. It was conducted in accordance with the French Good Epidemiological Practice guidelines.
6
Gaucher Disease hiPSC Macrophage Treatment
7
Fluorometric Assay for β-Glucocerebrosidase Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The inhibitory activity against β-glucocerebrosidase was measured using Cerezyme (Genzyme; Boston, MA, USA) as an enzyme source and 4-methylumbelliferyl-β-d -glucopyranoside (Sigma-Aldrich Co., St. Louis, MO, USA) as the substrate. The reaction mixture consisted of 100 mM McIlvaine buffer (pH 5.2), 0.25% sodium taurocholate, 0.1% Triton X-100 (Nacalai Tesque Inc., Kyoto, Japan), and the appropriate amount of enzyme. The reaction mixture was preincubated at 0 °C for 45 min; then 3 mM substrate solution was added, followed by incubation at 37 °C for 30 min. The reaction was stopped upon addition of 1.6 mL of a 400 mM glycine–NaOH solution (pH 10.6). The released 4-methylumbelliferone was measured (excitation wavelength = 362 nm; emission wavelength = 450 nm) using a F-4500 fluorescence spectrophotometer (Hitachi, Tokyo, Japan).
8
Velaglucerase Enzyme Replacement Therapy for GD1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 29 GD1 patients (18 female and 11 male) were included in this study. All patients were under enzyme replacement therapy (ERT) with velaglucerase (VPRIV, Shire), although some of them had previously received imiglucerase (Cerezyme, Genzyme). Average time of ERT with velaglucerase was 3.5 years (range: 1-5
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!