ABSTRACT
We have previously obtained compelling proof-of-principle evidence for COX2 gene therapy for fracture repair using integrating retroviral vectors. For this therapy to be suitable for patient uses, a suitable vector with high safety profile must be used. Accordingly, this study sought to evaluate the feasibility of AAV as the vector for this COX2 gene therapy, because AAV raises less safety issues than the retroviral vectors used previously. However, an appropriate AAV serotype is required to provide early increase in and adequate level of COX2 expression that is needed for fracture repair. Herein, we reported that AAV-DJ, an artificial AAV pseudoserotype, is highly effective in delivering COX2 gene to fracture sites in a mouse femoral fracture model. Compared with AAV-2, the use of AAV-DJ led to ~5-fold increase in infectivity in mesenchymal stem cells (MSCs) and provided an earlier and significantly higher level of transgene expression at the fracture site. Injection of this vector at a dose of 7.5 × 10(11) genomic copies led to high COX2 level at the fracture site on day 3 after injections and significantly promoted fracture union at 21 days, as analyzed by radiography and µ-CT. The therapeutic effect appears to involve enhanced osteoblastic differentiation of MSCs and remodeling of callus tissues to laminar bone. This interpretation is supported by the enhanced expression of several key genes participating in the fracture repair process. In conclusion, AAV-DJ is a promising serotype for the AAV-based COX2 gene therapy of fracture repair in humans.
Subject(s)
Cyclooxygenase 2/metabolism , Dependovirus/metabolism , Fracture Healing , Tibia/injuries , Transgenes , Animals , Disease Models, Animal , Genetic Therapy , Genetic Vectors/administration & dosage , Male , Mice, Inbred C57BLABSTRACT
Multiple drug resistance is a common problem in the treatment of epilepsy, and approximately 30% of patients continue to have seizures despite all therapeutic interventions. Among various classes of drug transporters, genetic variants of P-glycoprotein (P-gp) encoded by the ABCB1 (ATP-binding cassette subfamily B member 1) gene have been associated with drug-refractory epilepsy. Our aim was to investigate the effect of the 1236C>T(rs1128503), 2677G>T/A(rs2032582), and 3435C>T(rs1045642) single-nucleotide polymorphisms of ABCB1 (or MDR1) on drug resistance in north Indian patients with epilepsy. Genotyping was performed in 101 control subjects and 325 patients with epilepsy, of whom 94 were drug resistant and 231 drug responsive. Therapeutic drug monitoring for phenytoin, carbamazepine, phenobarbital, and valproate was also performed to confirm compliance in 20% of the patients. Genotype and haplotype frequencies of these polymorphisms did not differ between drug-resistant and drug-responsive patients. Our results demonstrate ABCB1 polymorphisms are not associated with drug resistance in north Indian epileptic patients.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Anticonvulsants/therapeutic use , Epilepsy/epidemiology , Epilepsy/genetics , Polymorphism, Genetic/genetics , ATP Binding Cassette Transporter, Subfamily B , Adult , Alleles , DNA/genetics , Drug Resistance/genetics , Epilepsy/drug therapy , Female , Gene Frequency , Genetic Variation , Genotype , Haplotypes , Humans , India/epidemiology , Male , Polymorphism, Single Nucleotide , Reverse Transcriptase Polymerase Chain Reaction , Sex Factors , Young AdultABSTRACT
The synthesis of fifteen new 1-aryl-2-amino-3-(4-arylthiazol-2-yl)/(benzothiazol-2-yl)gua nidines is described. They were screened for their antimicrobial susceptibility by the standard disc diffusion method of the World Health Organization (WHO) and the activities compared with that of standard strain of Escherichia coli NCTC 10418. The sensitive aminoguanidines were further subjected to the minimum inhibitory concentration (MIC) test.
Subject(s)
Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/pharmacology , Guanidines/chemical synthesis , Guanidines/pharmacology , Anti-Bacterial Agents/chemistry , Bacteria/drug effects , Guanidines/chemistry , Magnetic Resonance Spectroscopy , Microbial Sensitivity Tests , Molecular StructureABSTRACT
A series of p-nitrophenyl substituted semicarbazones has been synthesised and their anticonvulsant activity was screened against MES, scPTZ and scSTY. 4(4'-Nitrophenyl)-o-nitrobenzaldehyde semicarbazone has been found to be the most active in all these tests. The studies revealed that a primary amino function is not essential for anticonvulsant activity in the semicarbazone series of compounds. Presumably these compounds could further act on glycine receptors.
Subject(s)
Anticonvulsants/chemical synthesis , Semicarbazones/chemical synthesis , Animals , Anticonvulsants/pharmacology , Chemical Phenomena , Chemistry, Physical , Convulsants , Electroshock , Magnetic Resonance Spectroscopy , Mice , Pentylenetetrazole , Postural Balance/drug effects , Seizures/chemically induced , Seizures/prevention & control , Semicarbazones/pharmacology , Spectrophotometry, Infrared , StrychnineABSTRACT
Some new 2-carbamoylmethylthio- and 2-(omega-dimethylamino-alkyl)thio-3-aryl-7-chloro-4(3H)-quinazolinones have been prepared from 2-thio-3-aryl-7-chloro-4(3H)-quinazolinones as the key intermediate. Five of the synthetic compounds were screened for their CNS activities on mice and found to be either CNS depressants or stimulants.
Subject(s)
Central Nervous System Agents/chemical synthesis , Quinazolines/chemical synthesis , Animals , Mice , Quinazolines/pharmacology , Structure-Activity RelationshipABSTRACT
Synthesis is described of eleven new 2-(N-substituted or N,N-disubstituted aminoacetamido)-4- or -4,5-substituted thiazoles (I). Their hydrochlorides were screened for local anaesthetic activity on frogs by the sciatic plexus method and the activity compared with that of procaine.
