Mohammad Farhadi

GJB2 MUTATIONS: PASSAGE THROUGH IRAN

Hereditary hearing loss (HHL) is a very common disorder. When inherited in an autosomal recessive manner, it typically presents as an isolated finding. Interestingly and unexpectedly, in spite of extreme heterogeneity, mutations in one gene, GJB2, are the most common cause of congenital severe‐to‐profound deafness in many different populations. In this study, we assessed the contributions made by GJB2 mutations and chromosome 13 g.1777179_2085947del (the deletion more commonly known as del (GJB6‐D13S1830) that includes a portion of GJB6 and is hereafter called Δ(GJB6‐D13S1830)) to the autosomal recessive non‐syndromic deafness (ARNSD) genetic load in Iran. Probands from 664 different nuclear families were investigated. GJB2‐related deafness was found in 111 families (16.7%). The carrier frequency of the 35delG mutation showed a geographic variation that is supported by studies in neighboring countries. Δ(GJB6‐D13S1830) was not found. Our prevalence data for GJB2‐related deafness reveal a geographic pattern that mirrors the south‐to‐north European gradient and supports a founder effect in southeastern Europe.

A mutation in HOXA2 is responsible for autosomal-recessive microtia in an Iranian family.

Microtia, a congenital deformity manifesting as an abnormally shaped or absent external ear, occurs in one out of 8,000-10,000 births. We ascertained a consanguineous Iranian family segregating with autosomal-recessive bilateral microtia, mixed symmetrical severe to profound hearing impairment, and partial cleft palate. Genome-wide linkage analysis localized the responsible gene to chromosome 7p14.3-p15.3 with a maximum multi-point LOD score of 4.17. In this region, homeobox genes from the HOXA cluster were the most interesting candidates. Subsequent DNA sequence analysis of the HOXA1 and HOXA2 homeobox genes from the candidate region identified an interesting HOXA2 homeodomain variant: a change in a highly conserved amino acid (p.Q186K). The variant was not found in 231 Iranian and 109 Belgian control samples. The critical contribution of HoxA2 for auditory-system development has already been shown in mouse models. We built a homology model to predict the effect of this mutation on the structure and DNA-binding activity of the homeodomain by using the program Modeler 8v2. In the model of the mutant homeodomain, the position of the mutant lysine side chain is consistently farther away from a nearby phosphate group; this altered position results in the loss of a hydrogen bond and affects the DNA-binding activity.

Biological activity of camel milk casein following enzymatic digestion

The aim of this study was to investigate the effects of enzymatic hydrolysis with digestive enzymes of camel whole casein and beta-casein (β-CN) on their antioxidant and Angiotensin Converting Enzyme (ACE)-inhibitory properties. Peptides in each hydrolysate were fractionated with ultra-filtration membranes. The antioxidant activity was determined using a Trolox equivalent antioxidant capacity (TEAC) scale. After enzymatic hydrolysis, both antioxidant and ACE-inhibitory activities of camel whole casein and camel β-CN were enhanced. Camel whole casein and β-CN showed significant ACE-inhibitory activities after hydrolysis with pepsin alone and after pepsinolysis followed by trypsinolysis and chymotrypsinolysis. Camel β-CN showed high antioxidant activity after hydrolysis with chymotrypsin. The results of this study suggest that when camel milk is consumed and digested, the produced peptides start to act as natural antioxidants and ACE-inhibitors.

Corticosteroid‐releasing cochlear implant: A novel hybrid of biomaterial and drug delivery system

In this study, drug-eluting cochlear implant (CI) electrodes were prepared, and the amount of drug released was determined. Dexamethasone (DEX) (0.25-2% w/w, the weight percent of the final cured polymer) was used as a bioactive agent to suppress postsurgical inflammations upon mixing with a two-part nonrestricted pourable medical-grade silicone elastomer. Batch reproducibility analysis was performed on three consecutive batches. Drug release experiments were accomplished in normal saline medium, where DEX was analyzed via a validated HPLC method. The drug loading percentage and the device surface area were the most dominant parameters explored to monitor the drug release behavior from CI coatings. Total cumulative amount of DEX released from various loaded samples was in the order of 2 > 1 > 0.5 > 0.35 > 0.25% w/w, but the cumulative percentage of drug released showed a reverse order. The DEX dosages between 0.1 and 1 microg were released from samples of smallest to highest loadings during the initial 24 h, and dosages <1-5 microg were released from similar samples of various loadings at the first patency 2 weeks. The extent of crosslinking was only effective on release profile at lower drug loadings of 0.25% w/w relative to 0.5%. It was also found that release profile was not affected by postcuring. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010.

Dexamethasone eluting cochlear implant: Histological study in animal model

Abstract Objective New cochlear implant (CI) designs and developments in implantation techniques have revolutionized the management of hearing loss. However, cochlear implantation still has some disadvantages, such as its potential to initiate an inflammatory response that may lead to further hair cell damage. Recent topics of investigation have been the effect of glucocorticoids on inflammatory tissue response reduction, glucocorticoid dosage levels, and drug-delivery methods. In the present study, dexamethasone delivery via a drug-eluting CI was evaluated histologically through assessing inflammatory cell infiltration. Methods and materials Thirty healthy, adult male guinea pigs were included and randomly assigned to one of three surgical groups that underwent cochleostomy of the basal turn. The experimental group (Group 1) of 12 animals were implanted with a dexamethasone-loaded silicone elastomer shaped like a CI electrode. The primary control group (Group 2) of 12 animals were implanted with a simple CI (non-eluting). A second control group (Group 3) of six animals underwent cochleostomy only. Inflammatory responses were compared between groups by evaluating inflammatory cell infiltration in inner-ear specimens at days 3 and 13. Results The Mann–Whitney test revealed reduction in most of the inflammatory indices in Group 1 compared with Group 2. This was significant for fibrocyte, macrophage, and giant cell infiltration at day 3 as well as lymphocyte, macrophage infiltration, and capillary formation at day 13. Conclusion This study showed some attenuation in inflammatory response following insertion of a dexamethasone-eluting CI, suggesting that it could be a route for local drug delivery into the cochlea.

Functional brain abnormalities localized in 55 chronic tinnitus patients: fusion of SPECT coincidence imaging and MRI

Tinnitus is often defined as the perception of sounds or noise in the absence of any external auditory stimuli. The pathophysiology of subjective idiopathic tinnitus remains unclear. The aim of this study was to investigate the functional brain activities and possible involved cerebral areas in subjective idiopathic tinnitus patients by means of single photon emission computerized tomography (SPECT) coincidence imaging, which was fused with magnetic resonance imaging (MRI). In this cross-sectional study, 56 patients (1 subject excluded) with subjective tinnitus and 8 healthy controls were enrolled. After intravenous injection of 5 mCi F18-FDG (fluorodeoxyglucose), all subjects underwent a brain SPECT coincidence scan, which was then superimposed on their MRIs. In the eight regions of interest (middle temporal, inferotemporal, medial temporal, lateral temporal, temporoparietal, frontal, frontoparietal, and parietal areas), the more pronounced values were represented in medial temporal, inferotemporal, and temporoparietal areas, which showed more important proportion of associative auditory cortices in functional attributions of tinnitus than primary auditory cortex. Brain coincidence SPECT scan, when fused on MRI is a valuable technique in the assessment of patients with tinnitus and could show the significant role of different regions of central nervous system in functional attributions of tinnitus.

Characterising the spectrum of autosomal recessive hereditary hearing loss in Iran

Background Countries with culturally accepted consanguinity provide a unique resource for the study of rare recessively inherited genetic diseases. Although hereditary hearing loss (HHL) is not uncommon, it is genetically heterogeneous, with over 85 genes causally implicated in non-syndromic hearing loss (NSHL). This heterogeneity makes many gene-specific types of NSHL exceedingly rare. We sought to define the spectrum of autosomal recessive HHL in Iran by investigating both common and rarely diagnosed deafness-causing genes. Design Using a custom targeted genomic enrichment (TGE) panel, we simultaneously interrogated all known genetic causes of NSHL in a cohort of 302 GJB2-negative Iranian families. Results We established a genetic diagnosis for 67% of probands and their families, with over half of all diagnoses attributable to variants in five genes: SLC26A4, MYO15A, MYO7A, CDH23 and PCDH15. As a reflection of the power of consanguinity mapping, 26 genes were identified as causative for NSHL in the Iranian population for the first time. In total, 179 deafness-causing variants were identified in 40 genes in 201 probands, including 110 novel single nucleotide or small insertion–deletion variants and three novel CNV. Several variants represent founder mutations. Conclusion This study attests to the power of TGE and massively parallel sequencing as a diagnostic tool for the evaluation of hearing loss in Iran, and expands on our understanding of the genetics of HHL in this country. Families negative for variants in the genes represented on this panel represent an excellent cohort for novel gene discovery.

Inhibition study on insulin fibrillation and cytotoxicity by paclitaxel.

Alzheimer, a neurodegenerative disease, and a large variety of pathologic conditions are associated with a form of protein aggregation known as amyloid fibrils. Since fibrils and prefibrillar intermediates are cytotoxic, numerous attempts have been made to inhibit fibrillation process as a therapeutic strategy. Peptides, surfactants and aromatic small molecules have been used as fibrillation inhibitors. Here we studied the effects of paclitaxel, a polyphenol with a high tendency for interaction with proteins, on fibrillation of insulin as a model protein. The effects of paclitaxel on insulin fibrillation were determined by Thioflavin T fluorescence, Congo red absorbance, circular dichroism and atomic force microscopy. These studies indicated that paclitaxel considerably hindered nucleation, and therefore, fibrillation of insulin in a dose-dependant manner. The isothermal titration calorimetry studies showed that the interaction between paclitaxel and insulin was spontaneous. In addition, the van der Waals interactions and hydrogen bonds were prominent forces contributing to this interaction. Computational results using molecular dynamic simulations and docking studies revealed that paclitaxel diminished the polarity of insulin dimer and electrostatic interactions by increasing the hydrophobicity of its dimer state. Furthermore, paclitaxel reduced disrupting effects of insulin fibrils on PC12 cells neurite outgrowth and complexity, and enhanced their survival.

Hemoglobin fructation promotes heme degradation through the generation of endogenous reactive oxygen species

Protein glycation is a cascade of nonenzymatic reactions between reducing sugars and amino groups of proteins. It is referred to as fructation when the reducing monosaccharide is fructose. Some potential mechanisms have been suggested for the generation of reactive oxygen species (ROS) by protein glycation reactions in the presence of glucose. In this state, glucose autoxidation, ketoamine, and oxidative advance glycation end products (AGEs) formation are considered as major sources of ROS and perhaps heme degradation during hemoglobin glycation. However, whether fructose mediated glycation produces ROS and heme degradation is unknown. Here we report that ROS (H2O2) production occurred during hemoglobin fructation in vitro using chemiluminescence methods. The enhanced heme exposure and degradation were determined using UV-Vis and fluorescence spectrophotometry. Following accumulation of ROS, heme degradation products were accumulated reaching a plateau along with the detected ROS. Thus, fructose may make a significant contribution to the production of ROS, glycation of proteins, and heme degradation during diabetes.

A scientific paradigm for targeted nanophotothermolysis; the potential for nanosurgery of cancer

The application of gold nanoparticles (AuNPs) in nanophotothermolysis as a great photosensitizer is expanding, and this subject is a challenging area for cancer therapy. Recent technological advances in nanoscale manufacturing and synthesis promise the development of highly beneficial and innovative methods for the targeting of cancer. However, there is an obstacle to conducting effective laser-based nanosurgery because AuNPs are activated by visible or near infrared wavelengths, and the penetration of a laser beam inside the body is limited by some absorbents, such as melanin, water, and blood molecules. Considering everything stated above, we have suggested the application of a folate-conjugated AuNP as an effective agent for targeted nanophotothermolysis and the application of an optical fiber to transport the laser light from the source to the target tissue inside the body. Thus, a new method of nanosurgery in which a surgeon is able to perform surgery at the cellular or even at the subcellular level may be possible.