Genetic study of the PAH locus in the Iranian population: familial gene mutations and minihaplotypes

Phenylketonuria (PKU), one of the most common inborn errors of amino acid metabolism, is caused by mutations in the phenylalanine hydroxylase (PAH) gene (PAH). PKU has wide allelic heterogeneity, and over 600 different disease-causing mutations in PAH have been detected to date. Up to now, there have been no reports on the minihaplotype (VNTR/STR) analysis of PAH locus in the Iranian population. The aims of the present study were to determine PAH mutations and minihaplotypes in Iranian families with PAH deficiency and to investigate the correlation between them. A total of 81 Iranian families with PAH deficiency were examined using PCR-sequencing of all 13 PAH exons and their flanking intron regions to identify sequence variations. Fragment analysis of the PAH minihaplotypes was performed by capillary electrophoresis for 59 families. In our study, 33 different mutations were found accounting for 95% of the total mutant alleles. The majority of these mutations (72%) were distributed across exons 7, 11, 2 and their flanking intronic regions. Mutation c.1066-11G > A was the most common with a frequency of 20.37%. The less frequent mutations, p.Arg261Gln (8%), p.Arg243Ter (7.4%), p.Leu48Ser (7.4%), p.Lys363Asnfs*37 (6.79%), c.969 + 5G > A (6.17%), p.Pro281Leu (5.56), c.168 + 5G > C (5.56), and p.Arg261Ter (4.94) together comprised about 52% of all mutant alleles. In this study, a total of seventeen PAH gene minihaplotypes were detected, six of which associated exclusively with particular mutations. Our findings indicate a broad PAH mutation spectrum in the Iranian population, which is consistent with previous studies reporting a wide range of PAH mutations, most likely due to ethnic heterogeneity. High prevalence of c.1066-11G > A mutation linked to minihaplotype 7/250 among both Iranian and Mediterranean populations is indicative of historical and geographical links between them. Also, strong association between particular mutations and minihaplotypes could be useful for prenatal diagnosis (PND) and preimplantation genetic diagnosis (PGD) in affected families.

     

Effects of patellar taping on knee pain,functional disability,and patellar alignments in patients with patellofemoral pain syndrome: A randomized clinical trial

Question

What are the effects of patellar taping on pain, functional disability and patellar alignments in Patellofemoral Pain Syndrome (PFPS)?

Synthetic ionophores as non-resistant antibiotic adjuvants

Antimicrobial resistance is a world-wide health care crisis. New antimicrobials must both exhibit potency and thwart the ability of bacteria to develop resistance to them. We report the use of synthetic ionophores as a new approach to developing non-resistant antimicrobials and adjuvants. Most studies involving amphiphilic antimicrobials have focused on either developing synthetic amphiphiles that show ion transport, or developing non-cytotoxic analogs of such peptidic amphiphiles as colistin. We have rationally designed, prepared, and evaluated crown ether-based synthetic ionophores (‘hydraphiles’) that show selective ion transport through bilayer membranes and are toxic to bacteria. We report here that hydraphiles exhibit a broad range of antimicrobial properties and that they function as adjuvants in concert with FDA-approved antibiotics against multi-drug resistant (MDR) bacteria. Studies described herein demonstrate that benzyl C₁₄ hydraphile (BC₁₄H) shows high efficacy as an antimicrobial. BC₁₄H, at sub-MIC concentrations, forms aggregates of ∼200 nm that interact with the surface of bacteria. Surface-active BC₁₄H then localizes in the bacterial membranes, which increases their permeability. As a result, antibiotic influx into the bacterial cytosol increases in the presence of BC_nHs. Efflux pump inhibition and accumulation of substrate was also observed, likely due to disruption of the cation gradient. As a result, BC₁₄H recovers the activity of norfloxacin by 128-fold against resistant Staphylococcus aureus. BC₁₄H shows extremely low resistance development and is less cytotoxic than colistin. Overall, synthetic ionophores represent a new scaffold for developing efficient and non-resistant antimicrobial-adjuvants.

Antimicrobial resistance is a world-wide health care crisis.     

Amyloid-Beta: A Crucial Factor in Alzheimer's Disease

Alzheimer''s disease (AD) is the most prevalent form of dementia which affects people older than 60 years of age. In AD, the dysregulation of the amyloid-beta (Aβ) level leads to the appearance of senile plaques which contain Aβ depositions. Aβ is a complex biological molecule which interacts with many types of receptors and/or forms insoluble assemblies and, eventually, its nonphysiological depositions alternate with the normal neuronal conditions. In this situation, AD signs appear and the patients experience marked cognitional disabilities. In general, intellect, social skills, personality, and memory are influenced by this disease and, in the long run, it leads to a reduction in quality of life and life expectancy. Due to the pivotal role of Aβ in the pathobiology of AD, a great deal of effort has been made to reveal its exact role in neuronal dysfunctions and to finding efficacious therapeutic strategies against its adverse neuronal outcomes. Hence, the determination of its different molecular assemblies and the mechanisms underlying its pathological effects are of interest. In the present paper, some of the well-established structural forms of Aβ, its interactions with various receptors and possible molecular and cellular mechanisms underlying its neurotoxicity are discussed. In addition, several Aβ-based rodent models of AD are reviewed.Key Words: Alzheimerߣs disease, Amyloid-beta, Pathobiology     

Conjugated Linoleic Acid Stimulates Apoptosis in RH and Tehran Strains of Toxoplasma gondii,in Vitro

Background:

The aim of this study was to evaluate the effects of conjugated linoleic acid (CLA) on apoptosis of tachyzoites of T. gondii, RH strain (type I) and the cyst-forming Tehran strain (type II) in vitro.

Methods:

Toxoplasma strains were injected into the peritoneal cavity of BALB/c mice. The Tehran strain forms cysts in the brain of mice. Bradyzoites within the cysts are reactivated to proliferative tachyzoites, by dexamethasone. Tachyzoites were aspirated from the peritoneum of infected mice, and the percentage of viable parasites was estimated with trypan blue staining. Tachyzoites were inoculated into HeLa cells cultivated in DMEM medium. Different concentrations of CLA were evaluated on T. gondii in HeLa cells by the tetrazolium (MTT) colorimetric assay. Differentiation between apoptosis and cell death was determined by flow cytometry using Annexin V and propidium iodide (PI) double staining. The statistical analysis performed by GraphPad Prism version 6.00.

Results:

CLA induces apoptosis in virulent (RH) and avirulent (Tehran) strains of T. gondii. The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.

Conclusion:

Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzoites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.     

Comparison between Combination Therapy of Oral Terbinafine and Cryotherapy versus Systemic Meglumine Antimoniate and Cryotherapy in Cutaneous Leishmaniasis: A Randomized Clinical Trial

Background:

Leishmaniasis is a parasitic infection that may lead to a variety of manifestations. In Iran, cutaneous leishmaniasis (CL) has a high prevalence. There are many treatment modalities for CL. The use of oral terbinafine in the treatment of CL has recently been considered. The aim of this study was to compare combination of oral terbinafine plus cryotherapy versus systemic meglumine antimoniate plus cryotherapy in CL.

Methods:

Patients with proven direct smear for CL were divided randomly in 2 groups of 40 cases. For the first group systemic glucantime prescribed (IM, 15 mg/kg/day) for 3 weeks. For the second group oral terbinafine as two folds of usual dose in the treatment of fungal diseases prescribed [125 mg/day for body weight (BW) <20 kg, 250 mg/day for BW 20–40 kg, 500 mg/day for BW>40 kg] for 4 weeks. Both groups received cryotherapy every 2 weeks for 4 weeks. The patients were followed monthly for 3 months after the treatment.

Results:

Partial (HR= 0.55, CI 95%= 0.3–1.1) and complete (HR= 0.53, CI 95%= 0.3–0.98) clinical improvement in terbinafine group was much slower than glucantime group, although at the end of treatment protocols no significant difference between groups were statistically observed (P=0.27).

Conclusion:

Considering more convenient suitable route of administration and approximately comparable results, it seems that terbinafine can be used as an alternative treatment, especially in the case of allergy or resistance to systemic glucantime.     

A comparison of performance of anti-cyclic citrullinated peptide 2 and citrullinated protein antibodies in the diagnosis of rheumatoid arthritis in Iranian patients

Antibodies to citrullinated proteins and rheumatoid factor (RF) are widely used in patients with rheumatoid arthritis (RA) and the antibodies to citrullinated proteins appear to be the most specific markers of the disease. The objective was to compare the diagnostic performance of the anti-cyclic citrullinated peptide 2 (anti-CCP2) and citrullinated protein Antibodies (CPA) with RF in the diagnosis of RA. Serum samples of 139 patients with RA and 131 patients with other rheumatic diseases were checked for anti-CCP2, CPA uses citrullinated recombinant rat filaggrin as the antigen assay, and RF. The specificity, sensitivity, and receiver operating characteristic (ROC) of tests were then compared. The sensitivity of anti-CCP2, CPA, and RF were 82.7, 83.5, and 61.5%, respectively. The specificities of the tests were 91.2, 78.6, and 90.5%, respectively. The area under ROC curves for the tests were 0.925, 0.890, and 0.847, respectively. Exclusion of overlaps was associated with improved specificity for CPA but no change in the specificity of RF and anti-CCP. The sensitivity of anti-CCP2, CPA, and RF were 66.7, 77.8, and 51.9% for patients with early RA, respectively. The findings of the present study indicate that anti-CCP2 might be of a better diagnostic value for the diagnosis of RA. They also showed that CPA and in the second place anti-CCP2 were useful in the diagnosis of early RA.