Gaucher Disease: New Expanded Classification Emphasizing Neurological Features

 Gaucher disease (GD) is a rare inherited metabolic disorder and themost common lysosomal storage disorder, caused by a deficiency inglucocerebrosidase enzyme activity. It has been classified accordingto the neurological manifestations into three types: type 1, withoutneuropathic findings, type 2 with acute infantile neuropathic signsand type 3 or chronic neuropathic form. However, report of newvariants has led to the expansion of phenotype as a clinical phenotypeof GD considered as a continuum of phenotypes. Therefore, it seemsthat a new classification is needed to cover new forms of the disease. Keywords: Gaucher disease; Neurological manifestations; Phenotypes

Compressive Strength of Mineral Trioxide Aggregate with and without Disodium Hydrogen Phosphate at Different Mixing Ratios

Introduction: Mineral Trioxide Aggregate (MTA) is a substance with favorable physical-mechanical properties. Disodium hydrogen phosphate(DHP) is sometimes added to MTA to reduce its setting time. Therefore, this study was conducted to evaluate the effect of various ratios of liquid to powder of white MTA (WMTA) and addition of DHP on its compressive strength. Methods and Materials: One hundred and twenty samples were prepared with a two-piece stainless steel mold with a height of 6 mm and a diameter of 4 mm in order to evaluate the compressive strength where WMTA was used in 60 samples and DHP in white MTA composition (DHPWMTA) was used in other 60 samples. The compressive strength of WMTA and DHPWMTA was measured in various ratios of liquid to powder including 50, 60 and 70% and at 24 h and 21 days (n=10). Univariate Analysis of Variance test with SPSS 16 software were used to determine the difference between groups. The level of significance was set at 0.05. Results: The maximum and minimum compressive strength of WMTA groups were 63.25±1.96 (50% ratio and 21 days) and 37.79±1.28 (70% ratio and 24 h), respectively. The maximum and minimum compressive strength of DHPWMTA groups were 63.96±1.40 (60% ratio and 21 days) and 37.37±1.62 (70% ratio and 24 h), respectively. The effect of each of factors (type of material, powder to liquid ratio and time) alone were significant on the compressive strength (P<0.05). However, the interactive effect of three factors (type of material, powder to liquid ratio and time) were not statistically significant on compressive strength (P>0.05). Conclusion: Adding 2.5 wt% of DHP to white MTA increased samples compressive strength. Compressive strength in liquid to powder ratios of 50 and 60% compare to 70% and at 21 days compared to 24 h was high.Keywords: Compressive Strength; Disodium Hydrogen Phosphate; Mineral Trioxide Aggregat

Chitotriosidase Activity and Gene Polymorphism in Iranian Patients with Gaucher Disease and Sibling Carriers

How to Cite This Article: Mozafari H, Taghikhani M, Khatami Sh, Alaei MR, Vaisi-Raygani A, Rahimi Z. Chitotriosidase Activity and Gene Polymorphism in Iranian Patients with Gaucher Disease and Sibling Carriers. Iran J Child Neurol. Autumn 2016; 10(4):62-70.AbstractObjectiveChitotriosidase (CT) activity is a useful biomarker for diagnosis and monitoring of Gaucher disease (GD). Its application is limited by some variants in the CT gene. Two main polymorphisms are 24 bp duplication and G102S led to reduce CT activity. The aim of this study was to determine these variants influencing on plasma CT activity. Materials & MethodsBlood samples were collected from 33 patients with GD, 15 sibling carriers and 105 healthy individuals serving as controls. CT activity was measured using 4-methylumbelliferyl-β-D-N,N′,N″triacetylchitotrioside substrate in plasma samples. The CT genotypes of 24 bp duplication and G102S variants were determined using PCR and PCR-RFLP. ResultsUntreated GD patients had a significantly higher CT activity compared to treated patients (P = 0.021). In addition, chitotriosidase activity in carriers was higher rather than controls. Allele frequencies of 24 bp duplication in GD patients, sibling carriers and controls were 0.21, 0.266 and 0.29 and for G102S were 0.318, 0.366 and 0.219, respectively. Different G102S genotypes had not significant effect on CT activity. Chitotriosidase activity has a positive correlation with age in normal group, carriers, and negative correlation with hemoglobin in GD patients. Using cut-off level of 80.75 nmol/ml/h, sensitivity and specificity of CT activity were 93.9% and 100%, respectively. ConclusionChitotriosidase activity is a suitable biomarker for diagnosis and monitoring of GD. Determination of 24 bp duplication is helpful for more accurate monitoring the GD patient’s therapy. However, it seems that, specifying of the G102S polymorphism is not required for Iranian GD patients. References1. Bennett LL, Mohan D. Gaucher disease and its treatment  options. Ann Pharmacother 2013;47(9):1182-93.2. Shrestha B, Devgan A, Sharma M. Gaucher’s disease: rare presentation of a rare disease. J Child Neurol 2013;28(10):1296-8.3. Kanneganti M, Kamba A, Mizoguchi E. Role of chitotriosidase (chitinase 1) under normal and disease conditions. J Epithel Biol Pharmacol 2012;5:1-9.4. Adly AA, Ismail EA, Ibraheem TM. Macrophagederived soluble CD163 level in young patients with Gaucher disease: relation to phenotypes, disease severity and complications. Int Immunopharmacol 2015;24(2):416-22.5. Irún P, Alfonso P, Aznarez S, Giraldo P, Pocovi M.Chitotriosidase variants in patients with Gaucher disease.  Implications for diagnosis and therapeutic monitoring. Clin Biochem 2013;46(18):1804-7.6. Grace ME, Balwani M, Nazarenko I, Prakash- Cheng A, Desnick RJ. Type 1 Gaucher disease: null and hypomorphic novel chitotriosidase mutationsimplications for diagnosis and therapeutic monitoring. Hum Mutat 2007;28(9):866-73.7. Woo KH, Lee BH, Heo SH, Kim JM, Kim GH, Kim YM, et al. Allele frequency of a 24 bp duplication in exon 10 of the CHIT1 gene in the general Korean population and in Korean patients with Gaucher disease. J Hum Genet 2014;59(5):276-9.8. Wajner A, Michelin K, Burin MG, Pires RF, Pereira ML, Giugliani R, et al. Comparison between the biochemical properties of plasma chitotriosidase from normal individuals and from patients with Gaucher disease, GM1-gangliosidosis, Krabbe disease and heterozygotes for Gaucher disease. Clin Biochem 2007;40(5-6):365-9.9. Rosén C, Andersson CH, Andreasson U, Molinuevo JL, Bjerke M, Rami L, et al. Increased Levels of Chitotriosidase and YKL-40 in Cerebrospinal Fluid from Patients with Alzheimer’s Disease. Dement Geriatr Cogn Dis Extra 2014;31;4(2):297-304.10. Malaguarnera L. Chitotriosidase: the yin and yang. Cell Mol Life Sci 2006;63(24):3018-29.11. Pagliardini V, Pagliardini S, Corrado L, Lucenti A, Panigati L, Bersano E, et al. Chitotriosidase and lysosomal enzymes as potential biomarkers of disease progression in myotrophic lateral sclerosis: A survey clinic-based study. J Neurol Sci 2015;15;348(1-2):245-50.12. Fusetti F, von Moeller H, Houston D, Rozeboom HJ, Dijkstra BW, Boot RG, et al. Structure of human chitotriosidase. Implications for specific inhibitor design and function of mammalian chitinase-like lectins. J Biol Chem 2002;277:25537–25544.13. Sista RS, Wang T, Wu N, Graham C, Eckhardt A, Bali D, et al. Rapid assays for Gaucher and Hurler diseases in dried blood spots using digital microfluidics. Mol Genet Metab 2013;109(2): 218–220.14. Hollak CE, van Weely S, van Oers MH, Aerts JM. Marked elevation of plasma chitotriosidase activity. A novel hallmark of Gaucher disease. J Clin Invest 1994;93(3):1288–1292.15. Old JM, Higgs DR. Gene analysis. In: Weatherall DJ, editor. Methods in hematology. The thalassemias. Vol. 6. London: Churchill Livingstone; 1983. pp.74 – 101.16. Sinha S, Singh J, Jindal SK, Birbian N, Singla N. Association of 24 bp duplication of human CHIT1 gene with asthma in a heterozygous population of north India: a case-control study. Lung 2014;192(5):685-91.17. Manno N, Sherratt S, Boaretto F, Coico FM, Camus CE, Campos CJ, et al. High prevalence of chitotriosidase  deficiency in Peruvian Amerindians exposed to chitinbearing food and enteroparasites. Carbohydr Polym 2014;26;113:607-14.18. Adelino TE, Martins GG, Gomes AA, Torres AA, Silva DA, Xavier VD, et al. Biochemical and Molecular Chitotriosidase Profiles in Patients with Gaucher Disease Type 1 in Minas Gerais, Brazil: New Mutation in CHIT1 Gene. JIMD Rep 2013;9:85-91.19. van Dussen L, Hendriks EJ, Groener JE, Boot RG, Hollak CE, Aerts JM. Value of plasma chitotriosidase to assess non-neuronopathic Gaucher disease severity and progression in the era of enzyme replacement therapy. J Inherit Metab Dis 2014;37(6):991-1001.20. Weinreb NJ, Aggio MC, Andersson HC, Andria G, Charrow J, Clarke JT, et al. Gaucher disease type 1: revised recommendations on evaluations and monitoring for adult patients. Semin Hematol 2004;41:15–22.21. Czartoryska B, Tylki-Szymańska A, Górska D. Serum chitotriosidase activity in Gaucher patients on enzyme replacement therapy (ERT). Clin Biochem 1998;3(5):417-20.22. Arndt S1, Hobbs A, Sinclaire I, Lane AB. Chitotriosidase deficiency: a mutation update in an african population. JIMD Rep 2013;10:11-6.23. Lee P, Waalen J, Crain K, Smargon A, Beutler E. Human chitotriosidase polymorphisms G354R and A442V associated with reduced enzyme activity. Blood Cells Mol Dis 2007;39(3):353-60.24. Chien YH, Chen JH, Hwu WL. Plasma chitotriosidase activity and malaria. Clin Chim Acta 2005 ;353(1-2):215 25. Bussink AP, Verhoek M, Vreede J, Ghauharalivan der Vlugt K, Donker-Koopman WE, Sprenger RR, et al. Common G102S polymorphism in chitotriosidase differentially affects activity towards 4-methylumbelliferyl substrates. FEBS J 2009;276(19):5678-88.26. Aerts JM, Kallemeijn WW, Wegdam W, Joao Ferraz M, van Breemen MJ, Dekker N, et al. Biomarkers in the diagnosis of lysosomal storage disorders: proteins, lipids, and inhibodies. J Inherit Metab Dis 2011;34(3):605-19.27. Giraldo P, Cenarro A, Alfonso P, Pérez-Calvo JI, Rubio- Félix D, Giralt M, et al. Chitotriosidase genotype and plasma activity in patients type 1 Gaucher’s disease and their relatives (carriers and non carriers). Haematologica 2001;86(9):977-84.28. Pocovi M, Cenarro A, Civeira F, Torralba MA, Perez- Calvo JI, Mozas P, et al. Beta-glucocerebrosidase gene locus as a link for Gaucher’s disease and familial hypoalpha- lipoproteinaemia. Lancet 1998;351(9120):1919-23.29. Fluiter K, van der Westhuijzen DR, van Berkel TJ. In vitro regulation of scavenger receptor BI and the selective uptake of high density lipoprotein choles-teryl esters in rat liver parenchymal and Kupffer cells. J Biol Chem 1998; 273:8434-8.30. Ries M, Schaefer E, Lührs T, Mani L, Kuhn J, Vanier MT, et al. Critical assessment of chitotriosidase analysis in the rational laboratory diagnosis of children with Gaucher disease and Niemann-Pick disease type A/B and C. J Inherit Metab Dis 2006;29:647–652.31. Kurt I, Abasli D, Cihan M, Serdar MA, Olgun A, Saruhan E, et al. Chitotriosidase Levels in Healthy Elderly Subjects. Ann N Y Acad Sci 2007;1100:185-8.32. Tamanaha P, D’Almeida V, Calegare BF, Tomita LY, Bittencourt LR, Tufik S. 24 bp duplication of CHIT1 gene and determinants of human chitotriosidase activity among participants of EPISONO, a population-based cross-sectional study, São Paulo, Brazil. Clin Biochem 2013;46(12):1084-8.33. Dodelson de Kremer R, Paschini de Capra A, Angaroni CJ, Giner de Ayala A. Plasma chitotriosidase activity in Argentinian patients with Gaucher disease, various lysosomal diseases and other inherited metabolic disorders. Medicina (B Aires). 1997;57(6):677-84.34. Goldim MP, Garcia Cda S, de Castilhos CD, Daitx VV, Mezzalira J, Breier AC, et al. Screening of high-risk Gaucher disease patients in Brazil using miniaturized dried blood spots and leukocyte techniques. Gene 2012;508(2):197-8.

Association between The SIRT1 and SIRT3 Levels and Gene Polymorphisms with Infertility in War Zones of Kermanshah Province, Iran: A Case-Control Study

Objective: War toxin, mustard gas, alkylating agent results in male infertility via inducing reactive oxygen species (ROS) production and DNA mutagenesis. SIRT1 and SIRT3 are multifunctional enzymes that involve in the DNA repair, oxidative stress responses. This study aim is to assess the correlation between serum levels of SIRT1, SIRT3 and both rs3758391T>C and rs185277566C>G gene polymorphisms with infertility in the war zones of Kermanshah province, Iran.Materials and Methods: In this case-control study based on the semen analysis, samples were divided into two groups infertile (n=100) and fertile (n=100). High-performance liquid chromatography (HPLC) method was used to determine the malondialdehyde level, and also a sperm chromatin dispersion (SCD) test was used to evaluate the DNA fragmentation rate. Using the colorimetric assays, superoxide dismutase (SOD) activity was measured. SIRT1 and SIRT3 protein levels were determined by using ELISA. The genetic variants of SIRT1 rs3758391T>C, and SIRT3 rs185277566C>G, were detected by polymerase chain reaction-restriction fragment length (PCR-RFLP) technique.Results: Malondialdehyde (MDA) level and the percentage of DNA fragmentation were higher in infertile samples, but serum levels of SIRT1 and SIRT3, and SOD activity was lower in infertile compared to fertile samples (PC polymorphism, and CG+GG genotypes and the G allele from SIRT3 rs185277566C>G polymorphism could increase risk of infertility (P<0.05).Conclusion: The results of this study suggest that war toxins through the impact on genotypes, decreasing levels of SIRT1 and SIRT3 and increasing levels of oxidative stress, lead to defects in the concentration, motility and morphology of sperms and thus, infertility in men

Analysis of glucocerebrosidase gene mutations in Iranian patients with Gaucher disease: Identification of 6 novel mutations

Abstract Objective Gaucher disease (GD) is the most common autosomal recessive disorder of glycolipid storage. It results from mutations in the glucocerebrosidase (GBA) gene and leads to GBA deficiency. Different mutations are associated with different phenotypes in the three major types of GD. Materials &amp; Methods The spectrum of mutations in GBA gene in 26 unrelated patients with GD from different Iranian populations was determined by DNA sequencing, polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP), and amplification-refractory mutation system (ARMS) methods. An in silico analysis was also performedfor novel mutations Results Six new mutations were identified in this study. The newly detected mutations that could be theoretically harmful included p.I200T (c.599T&gt;C), p.H312D (c.934C&gt;G), p.L325S (c.974T&gt;C), p.L393V (c.1177C&gt;G), p.S439G (c.1315A&gt;G), and p.M455R (c.1365G&gt;A). Also, p.L483P, p.N409S, p.W420X, p.E379K, p.R398Q, p.N227S,p.R202Q, and p.D448H mutations were identified in the patients. Besides, two new complex mutations, namely, p.S439G/p.S439G+p. E379K/- and p.R202Q/p.R202Q+p.N227S/p.N227S, were detected. The most common GBA mutation in the population was p.L483P with an allele frequency of 32.7%, followed by p.N409S (19.2%). ConclusionThe present study detected six new mutations of GBA gene among GD patients. Two mutations (p.L483P andp.N409S) were especially common among Iranians; this finding can be used in implementing screening programs and understanding the molecular basis of GD

Zeolites for theranostic applications

Theranostic platforms bring about a revolution in disease management. During recent years, theranostic nanoparticles have been utilized for imaging and therapy simultaneously. Zeolites, because of their porous structure and tunable properties, which can be modified with various materials, can be used as a delivery agent. The porous structure of a zeolite enables it to be loaded and unloaded with various molecules such as therapeutic agents, photosensitizers, biological macromolecules, MRI contrast agents, radiopharmaceuticals, near-infrared (NIR) fluorophores, and microbubbles. Furthermore, theranostic zeolite nanocarriers can be further modified with targeting ligands, which is highly interesting for targeted cancer therapies