Four novel ARSA gene mutations with pathogenic impacts on metachromatic leukodystrophy: a bioinformatics approach to predict pathogenic mutations

Metachromatic leukodystrophy (MLD) disorder is a rare lysosomal storage disorder that leads to severe neurological symptoms and an early death. MLD occurs due to the deficiency of enzyme arylsulfatase A (ARSA) in leukocytes, and patients with MLD excrete sulfatide in their urine. In this study, the ARSA gene in 12 non-consanguineous MLD patients and 40 healthy individuals was examined using polymerase chain reaction sequencing. Furthermore, the structural and functional effects of new mutations on ARSA were analyzed using SIFT (sorting intolerant from tolerant), I-Mutant 2, and PolyPhen bioinformatics software. Here, 4 new pathogenic homozygous mutations c.585G>T, c.661T>A, c.849C>G, and c.911A>G were detected. The consequence of this study has extended the genotypic spectrum of MLD patients, paving way to a more effective method for carrier detection and genetic counseling

Two novel tyrosinase (TYR) gene mutations with pathogenic impact on oculocutaneous albinism type 1 (OCA1).

Oculocutaneous albinism (OCA) is a heterogeneous group of autosomal recessive disorders resulting from mutations of the tyrosinase (TYR) gene and presents with either complete or partial absence of pigment in the skin, hair and eyes due to a defect in an enzyme involved in the production of melanin. In this study, mutations in the TYR gene of 30 unrelated Iranian OCA1 patients and 100 healthy individuals were examined using PCR-sequencing. Additionally, in order to predict the possible effects of new mutations on the structure and function of tyrosinase, these mutations were analyzed by SIFT, PolyPhen and I-Mutant 2 software. Here, two new pathogenic p.C89S and p.H180R mutations were detected in two OCA1 patients. Moreover, the R402Q and S192Y variants, which are common non-pathogenic polymorphisms, were detected in 17.5% and 35% of the patients, respectively. The outcome of this study has extended the genotypic spectrum of OCA1 patients, which paves the way for more efficient carrier detection and genetic counseling

Effect of Mozart Music on Hippocampal Content of BDNF in Postnatal Rats

Introduction: It has shown that listening to Mozart music can potentiate spatial tasks in human; and reduce seizure attacks in epileptic patients. A few studies have reported the effects of prenatal plus postpartum exposure of mice to the Mozart music on brain-drived neurotrophic factor (BDNF) in the hippocampus. Here we investigated the effect of postpartum exposure to The Mozart music on BDNF concentration in the hippocampus of rat.Methods: Thirty male one day old newborn Wistar rats divided randomly in two equal experimental and control groups. Experimental group exposed to slow rhythm Mozart music (Mozart Sonata for two pianos KV 448, 6 hour per day; sound pressure levels, between 80 and 100 dB) for 60 successive days. The control group was kept in separate room with housing conditions like experimental group except music exposure. After 60 days the rats were euthanized and hippocampuses extracted; then the content of BDNF protein was measured using ELISA sandwich method. Results: Data analysis revealed that rats exposed to Mozart Sonata music had significantly increased BDNF content in the hippocampus as compared to control rats (P±0.01). The concentrations of BDNF were 86.30±2.26 and 94.60 ±6.22 ng/g wet weight in control and music exposure groups respectively.Discussion: Exposure to the Mozart music early in life can increase the BDNF concentration in the hippocampus in rats

Four Novel p.N385K, p.V36A, c.1033–1034insT and c.1417–1418delCT Mutations in the Sphingomyelin Phosphodiesterase 1 (SMPD1) Gene in Patients with Types A and B Niemann-Pick Disease (NPD)

Background: Types A and B Niemann-Pick disease (NPD) are autosomal-recessive lysosomal storage disorders caused by the deficient activity of acid sphingomyelinase due to mutations in the sphingomyelin phosphodiesterase 1 (SMPD1) gene. Methods: In order to determine the prevalence and distribution of SMPD1 gene mutations, the genomic DNA of 15 unrelated Iranian patients with types A and B NPD was examined using PCR, DNA sequencing and bioinformatics analysis. Results: Of 8 patients with the p.G508R mutation, 5 patients were homozygous, while the other 3 were heterozygous. One patient was heterozygous for both the p.N385K and p.G508R mutations. Another patient was heterozygous for both the p.A487V and p.G508R mutations. Two patients (one homozygous and one heterozygous) showed the p.V36A mutation. One patient was homozygous for the c.1033–1034insT mutation. One patient was homozygous for the c.573delT mutation, and 1 patient was homozygous for the c.1417–1418delCT mutation. Additionally, bioinformatics analysis indicated that two new p.V36A and p.N385K mutations decreased the acid sphingomyelinase (ASM) protein stability, which might be evidence to suggest the pathogenicity of these mutations. Conclusion: with detection of these new mutations, the genotypic spectrum of types A and B NPD is extended, facilitating the definition of disease-related mutations. However, more research is essential to confirm the pathogenic effect of these mutations

Intravenous Injection of Human Umbilical Cord Matrix Stem Cell (Wharton Jelly Stem Cell) Provides Functional Recovery in a Rat Model of Traumatic Brain Injury

Objective: This study was designed to examine the effects of human umbilical cord matrixstem cell (hUCMSC) administration in rats for 6 weeks after traumatic brain injury (TBI).Materials and Methods: Adult male Wistar rats (n = 30) were injured with controlled corticalimpact device and divided into three groups. The treatment group (n = 10) was injectedwith 2 × 106 hUCMSC intravenously, the vehicle group (n=10) received phosphate bufferedsaline (PBS) whereas the control group (n = 10) receive nothing. All injections wereperformed one day after injury into the tail veins of the rats. All cells were labelled withBrdu before injection. Evaluation of the neurological function of the rats was performedbefore and after injury using Neurological Severity Scores (NSS). The rats were sacrificed6 weeks after TBI and brain sections were stained using Brdu immunohistochemistry.Results: Statistically significant improvement in functional outcome was observed in thetreatment group compared with the control group (p < 0.01). This benefit was visible 1 weekafter TBI and persisted for six weeks (end of trial). Histological analysis showed that hUCMSCwere present in the lesion boundary zone at 6 weeks in all cell injected animals.Conclusion: Rats injected with hUCMSC after TBI survive for at least six weeks and showfunctional improvemnt

DNA sequencing result from Exon 1 of the <i>TYR</i> gene showing c.539 A>G mutation.

<p><b>A</b>: Normal Sequence from control and unaffected father. <b>B</b>: Sequence from patient 19 and an unaffected mother showing the c.539 A>G heterozygous mutation.</p

Statistical and Bioinformatics Analysis of two novel pathogenic mutations.

<p>Fisher's exact test using the SPSS was used for statistical analysis of novel mutations. A <i>p</i>-value of <0.05 is considered statistically significant. Novel mutations were analyzed by three computational methods PolyPhen 2 (benign/damaging), SIFT (tolerated/deleterious), and I-Mutant 2.0 (increase stability/decrease stability) for Bioinformatics analysis in order to predict the functional impact of novel amino acid changes.</p><p><i>p</i>-value: statistically significant (<i>p</i><0.05).</p><p>PolyPhen Prediction Score: benign ≤0.5; probably damaging (0.5<).</p><p>SIFT Prediction Score: deleterious (≤0.05); tolerated (≥0.05).</p><p>I-Mutant 2.0 Prediction: sign of DDG: decrease stability or increase stability.</p><p>Statistical and Bioinformatics Analysis of two novel pathogenic mutations.</p