Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol

Purpose: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larger cohort of primary open angle glaucoma (POAG) patients which included late-onset patients. We also aimed to set up a microarray based protocol for mutation screening with an intent of using the protocol in a future population level screening program. Methods: Sixty three POAG patients, nine affected family members, and thirty three previously genotyped primary congenital glaucoma (PCG) patients were included in the study. Clinical examination included slit lamp biomicroscopy, IOP measurement, gonioscopic evaluation, fundus examination, and measurement of perimetry. G61E, R368H, R390H, and R469W were screened by a protocol that included multiplexed allele specific amplification in the presence of a protease (PrASE), use of sequence tagged primers, and hybridization to generic arrays on microarray slides. The entire coding sequences of CYP1B1 and myocilin (MYOC) genes were sequenced in all individuals assessed by the microarray assay to carry a mutation. Intragenic single nucleotide polymorphism (SNP) haplotpes were determined for mutated alleles. Results: Genotypes assessed by the array-based PrASE methodology were in 100 concordance with sequencing results. Seven mutation carrying POAG patients (11.1) were identified, and their distribution was quite skewed between the juvenile-onset individuals (5/21) as compared to late-onset cases (2/42). Four of the seven mutation carrying Iranian patients harbored two mutated alleles. CYP1B1 mutated alleles in Iranian PCG and POAG patients shared common haplotypes. MYOC mutations were not observed in any of the patients. Conclusions: The PrASE approach allowed reliable simultaneous genotyping of many individuals. It can be an appropriate tool for screening common mutations in large sample sizes. The results suggest that CYP1B1 is implicated in POAG among Iranians, notably in the juvenile-onset form. Contrary to POAG patients studied in other populations, many mutation harboring Iranian patients carry two mutated alleles. We propose an explanation for this observation. © 2008 Molecular Vision

Nanotechnology and bio-functionalisation for peripheral nerve regeneration

There is a high clinical demand for new smart biomaterials, which stimulate neuronal cell proliferation, migration and increase cell-material interaction to facilitate nerve regeneration across these critical-sized defects. This article briefly reviews several up-to-date published studies using Arginine-Glycine-Aspartic acid peptide sequence, nanocomposite based on polyhedral oligomeric silsesquioxane nanoparticle and nanofibrous scaffolds as promising strategies to enhance peripheral nerve regeneration by influencing cellular behaviour such as attachment, spreading and proliferation. The aim is to establish the potent manipulations, which are simple and easy to employ in the clinical conditions for nerve regeneration and repair

The Effect of Low-Dose Tacrolimus on Recurrent Laryngeal Nerve Regeneration in Pigs: A Pilot Study

Objective:The functional neurotrophic effects of systemic tacrolimus in a sub-immunosuppressive regimen in the minipig model of laryngeal reinnervation were investigated. Methods:Right recurrent laryngeal nerve transection and phrenic-abductor branch of recurrent laryngeal nerve anastomosis were performed in two minipigs that were administered low-dose oral tacrolimus (0.125 mg/kg) for one month. Vocal cord abduction at four months was rated by two blinded expert assessors on a four-point Likert scale from poor (1) to complete (4), and compared with five fully matched historical control animals that received the same surgery but no other interventions. Results:Right vocal cord abduction was more complete in tacrolimus-treated animals than controls (mean abduction score 2.3 vs. 1.5, p=0.019). Conclusion:Tacrolimus may have an important role in laryngeal reinnervation following allograft transplantation, and low-dose regimens may have applications in cranial and peripheral nerve injuries or in the reinnervation of tissue-engineered laryngeal constructs, but further studies are required. [Arch Clin Exp Surg 2014; 3(2.000): 78-86

The Effect of Low-Dose Tacrolimus on Recurrent Laryngeal Nerve Regeneration in Pigs: A Pilot Study

Objective:The functional neurotrophic effects of systemic tacrolimus in a sub-immunosuppressive regimen in the minipig model of laryngeal reinnervation were investigated. Methods:Right recurrent laryngeal nerve transection and phrenic-abductor branch of recurrent laryngeal nerve anastomosis were performed in two minipigs that were administered low-dose oral tacrolimus (0.125 mg/kg) for one month. Vocal cord abduction at four months was rated by two blinded expert assessors on a four-point Likert scale from poor (1) to complete (4), and compared with five fully matched historical control animals that received the same surgery but no other interventions. Results:Right vocal cord abduction was more complete in tacrolimus-treated animals than controls (mean abduction score 2.3 vs. 1.5, p=0.019). Conclusion:Tacrolimus may have an important role in laryngeal reinnervation following allograft transplantation, and low-dose regimens may have applications in cranial and peripheral nerve injuries or in the reinnervation of tissue-engineered laryngeal constructs, but further studies are required. [Arch Clin Exp Surg 2014; 3(2.000): 78-86