فهرست مطالب

  • Volume:7 Issue:2, 2018
  • تاریخ انتشار: 1397/10/04
  • تعداد عناوین: 15
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  • Armin Mokhtariye , Lida Hagh, Nazari , Abdol, Reza Varasteh , Fatemeh Keyfi * Pages 119-128
    Lysosomal storage disorders (LSD) are a class of metabolic disturbance in which manifested by the accumulation of large molecules (complex lipids, glycoproteins, glycosaminoglycans, etc.) in lysosomes. LSDs have a wide range of clinical symptoms that may contain organ dysfunction, neurological and skeletal disorders. The first stage of diagnosis is clinically suspected by a physician. Next stage is enzyme activity assays including Fluorometry and MS/MS methods. These methods usually placed in newborn program screening. The second laboratory diagnostic stage is molecular examination (RFLP-PCR and ARMS-PCR, Mutations Scanning Methods, DNA sequencing, MLPA and NGS methods) that is confirmation of the enzyme assays. In this article, routine diagnostic methods for LSDs were discussed. The gold standard for enzyme activity assay and molecular diagnosis is TMS and NGS, respectively.
    Keywords: Diagnostic methods, Enzyme activity, Lysosomal storage disease, Molecular assay
  • Mojgan Mohammadi , Shahriar Dabiri , Hamid Reza Mollaei , Samira Rezaee Jouzdani , Maryam Amizadeh , Jamshid Esmailzadeh , Mohammad Reza Baneshi , Aliasghar Arabi Mianroodi * Pages 129-135
    Background
    Recent studies have shown interleukin 4 (IL-4) and 5 lipoxygenase (5-LO) to play an important role in development of nasal polyposis. Investigation into the genetic factors associated with allergic and non-allergic nasal polyposis has been examined for more than fifteen years. Despite these efforts, the genetic factors underlying the development of nasal polyposis have yet to be clearly understood. The current study examined the relationship between C-590T promoter polymorphisms of the IL-4 gene and the presence of nasal polyps. Additionally, we examined the levels of 5-LO expression in nasal polyp tissue and its association with the IL-4 promoter gene polymorphisms.
    Methods
    A total of 320 subjects were enrolled in the study, of which 256 were healthy controls and 64 were patients with nasal polyps. The Real-Time PCR HRM-based method was used to determine the genotypes of IL-4 C-590T. The expression of 5-LO within the 64 samples of nasal polyp tissue was determined by immunohistochemical staining to examine the association of 5-LO with the IL-4 C-590T genotype.
    Results
    Genetic analysis showed a significant difference in the frequencies of the IL-4 polymorphisms at C-590T in patients with nasal polyps as compared with controls (p<0.001). No significant difference was seen in the expression of 5-LO among genotypes in patients with nasal polyps (p=0.139).
    Conclusions
    The results suggest that the inheritance of TT and CT genotypes at the IL-4 C-590T promoter gene is associated with nasal polyps however, there is no association between the expression of 5-LO in nasal polyp tissues and IL-4 C-590T genotypes in patients with nasal polyps
    Keywords: Gene polymorphism, IL-4, IL-4 C-590T, Nasal polyposis, 5-LO
  • Milad Gholami , Sahar Bayat , Saba Manoochehrabadi , Hossein Pashaiefar , Mir Davood Omrani , Hasan Jalaeikhoo , Vahid Reza Yassaee , Mohammad Reza Ebrahimpour , Farkhondeh Behjati , Reza Mirfakhraie * Pages 136-141
    Background
    The pathogenicity of acute myeloid leukemia (AML) is highly influenced by genetic alterations, such as chromosomal abnormalities. Additionally, aberrations in the mechanisms involved in gene expression have been identified to have a role in the development of AML. Contradictory evidence has been reported concerning the expression of the CEBPA gene in AML patients. Additionally, investigation into the expression of the CEBPA-AS gene has yet to be explored in AML patients. The aim of the present study was to investigate the relationship between the expression of the CEBPA and CEBPA-AS genes and AML in Iranian patients.
    Methods
    Using quantitative real-time PCR, the expression of the CEBPA and CEBPA-AS genes was examined in the peripheral blood samples of 58 patients with de novo adult AML, and in 20 healthy controls.
    Results
    Overall, CEBPA expression analysis showed a significant up-regulation in AML patients compared with healthy controls. Interestingly, a significant up-regulation of CEBPA was detected in the male AML patients. Significant CEBPA over-expression was observed in M0 (p-value=0.0001), M3 (p-value= 0.012) and M4 (p-value= 0.000) FAB subtypes. Our data has also demonstrated that CEBPA expression is up-regulated in favorable (p-value= 0.006) and adverse (p-value= 0.042) cytogenetic risk groups. In addition, the expression of CEBPA was significantly increased in AML patients with an abnormal karyotype. Ectopic expression of CEBPA-AS was detected in seven of the AML patients.
    Conclusions
    Our study provides evidence for the up-regulation of CEBPA and the ectopic expression of CEBPA-AS in AML patients, suggesting that these two genes may play an important role in the pathogenesis of AML. The role of CEBPA and CEBPA-AS in AML patients should be further explored. This will offer potential opportunities for the development of novel treatment strategies
    Keywords: Acute Myeloid Leukemia, CEBPA, CEBPA-AS, de novo, Expression
  • Farnaz Mohajertehran *, Hossein Ayatollahi, Amir Hossein Jafarian, Kamran Khazaeni, Mohammad Soukhtanloo, Mohammad Taghi Shakeri, Nooshin Mohtasham Pages 142-149
    Background
    Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignancy that is associated with high morbidity and mortality. Salivary lactate dehydrogenase (LDH concentration), as an expression of cellular necrosis, may be a special marker of lesions that occur with changes in the integrity of the oral mucosa. This study was performed to determine the accuracy of salivary LDH as a clinical marker for HNSCC detection and to investigate the relationship between salivary LDH levels and tissue tumor detection.
    Methods
    The case group consisted of 44 HNSCC patients and the control group consisted of 44 healthy subjects. The stage and grade of HNSCC were determined, and the LDH levels in collected saliva samples were measured in all subjects. The expression of LDH in tumors and healthy tissue margins was evaluated via immunohistochemistry.
    Results
    The expression of LDH in the saliva of patients with HNSCC is significantly higher than that in the saliva of the healthy control group. The expression of salivary LDH in patients with oral squamous cell carcinoma (OSCC) is significantly higher than that in the other patients and healthy individuals in the control group. The levels of salivary LDH in patients with SCC of the tongue and lower oral cavity were significantly higher than those in other patients affected with SCC in other parts of the head and neck (P<0.01).
    Conclusions
    As this enzyme increases simultaneously in both tumoral tissues and saliva, it can serve as a useful diagnostic marker for the early diagnosis and prediction of HNSCC.
    Keywords: Biomarker, Early diagnosis, Head, neck squamous cell carcinoma (HNSCC), Lactate dehydrogenase (LDH), Saliva
  • Vahid Yaghoubi Naei*, Mojtaba Sankian , Malihe Moghadam , Narges Farshidi , Seyed Hasan Ayati , Fatemeh Hamid , Abdol, Reza Varasteh Pages 150-155
    Background
    Gamma irradiation is a form of processing with an array of applications in medical sciences such as microbial decontamination, viruses inactivation, cervical carcinoma and breast cancer treatment. One of the ways in which gamma irradiation has the potential to be used is in reducing the allergenicity of food allergens.
    Methods
    In the present study, pistachios were irradiated with either a 1, 10, or 100 kGy dose of gamma irradiation. The binding rate of mice and human antibodies to the allergens of the pistachio extracts were examined via Western blot analysis.
    Results
    Our findings show an inverse dose-response relationship between the binding rate of antibodies to the pistachio allergens and the gamma irradiation dose. Despite these promising findings, the results of our sensory evaluation indicate that gamma irradiation causes undesirable changes to the sensory characteristics of pistachios, especially at the dose of 100 kGy.
    Conclusions
    Gamma irradiation appears to be an effective method in reducing the allergenicity of pistachios. Thus, this form of processing has the potential to prevent adverse allergic reactions to the major pistachio allergens in sensitized subjects. However, further research must be dedicated to examining the dose sufficient in reducing allergencity, while maintaining adequate sensory quality for satisfactory consumption
    Keywords: Allergenicity, Gamma irradiation, Pistachio
  • Davood Mansury Mr, Kiarash Ghazvini , Saeid Amel Jamehdar , Ali Badiee Dr, Mohsen Tafaghodi , Amin Reza Nikpoor , Yousef Amini , Mahmoud Reza Jaafari * Pages 156-166
    Background
    Due to the ineffectiveness of the BCG vaccine, especially in adult pulmonary tuberculosis (TB), and variable efficacies against childhood forms of TB, developing an effective TB vaccine is a major priority in controlling this disease. The aim of this study was to evaluate the immunogenicity of a DOTAP liposome formulation containing a fusion protein (FP) containing Mycobacterium tuberculosis HspX, PPE44, and EsxV.
    Methods
    The FP was expressed in E. coli BL21 cells and confirmed by SDS-PAGE and Western blots. The FP was then encapsulated in various liposomal formulations. Afterwards, liposomal size, zeta potential, and encapsulation efficiency were evaluated. Mice were subcutaneously vaccinated on days 0, 14, and 28 with liposomes containing the FP. Two weeks after the last injection, IFN-γ, IL-4, IL-17, and IL-12 in spleen cell culture supernatants, and IgG2a, IgG1, and IgG2b titers in sera were measured.
    Results
    The FP concentration was 1mg/ml. The encapsulation efficiency of the liposomes varied from 69% in Lip (DOTAP/TDB/CHOL/FP) to 80% in Lip (DOTAP/CHOL/FP). The greatest IFN-γ and IL-12 levels were observed in BCG-primed mice that were boosted with Lip (DOTAP/CHOL/FP). In addition, IL-17 production was significantly greater in all groups than controls except in those that received histidine buffer and FP. The IgG2a/IgG1 ratios were greater in the Lip (DOTAP/TDB/CHOL/FP), Lip (DOTAP/CHOL/FP), Lip (DOTAP/CHOL), and BCG-primed and Lip (DOTAP/CHOL/FP)-boosted groups than in the other groups, indicating a cellular immune response.
    Conclusions
    The liposomes containing DOTAP combined with the fusion protein induced a Th1 response. The mice that first received BCG and then Lip (DOTAP/CHOL/FP), produced the most IFN-γ and IL-12, indicating a strong Th1 response
    Keywords: DOTAP, Vaccines, Fusion Protein, Liposomes, Mycobacterium tuberculosis, Trehalose 6, 6’-Dibehenate
  • Akbar Derakhshan , Jalil Tavakkol Afshari , Javad Sadeghi Allah Abadi*, Amin Reza Nikpoor , Ramin Daneshvar , Saeed Shokoohi Rad , Mohammad, Reza Ansari, Astaneh Pages 167-173
    Background
    Glaucoma is a common cause of irreversible blindness. Transforming growth factor beta-1(TGF-β1) is the main isoform of TGF-β superfamily in the eye. Overexpression of TGF-β1 is shown to be related with the glaucoma. Studies have shown that the presence of mutant T allele of TGF-β1 -509C>T polymorphism (rs1800469) is associated with increased gene expression. So, in present study, association of the TGF-β1-509C>T gene polymorphism and primary open angle glaucoma (POAG) in patients from north east of Iran was investigated.
    Methods
    A case-control study was conducted on 112 POAG patients and 112 control participants. TGF-β1-509C>T genotyping was done by PCR-restriction fragment length polymorphism (PCR-RFLP) method using Bsu36I restriction enzyme. Moreover, cup to disk ratio(CDR), intraocular pressure (IOP) and visual acuity (VA) were measured. The obtained results were statistically analyzed.
    Results
    The highest frequency of genotype in the control group was related to CC genotype (44.6%), but the heterozygous CT genotype (45.6%) was observed as the highest frequency of genotypes in patient group (P value: 0.022, OR for TT genotype: 2.54 CI95% for OR: 1.22, 5.27). Also, the frequency of the T mutant allele showed a significant difference between case and control groups (P value: 0.005, OR: 1.73 CI95% for OR: 1.18, 2.53).
    Conclusions
    In conclusion, a significant association was seen between TGF-β1 -509C>T gene polymorphism and POAG disease and inheritance of mutant T allele is considered to be a risk factor for glaucoma in patients living in North Eastern part of Iran
    Keywords: Glaucoma, PCR, TGF-beta1, Polymorphism
  • Shobha Ullas Kamath , Abhishek Chaturvedi*, Devesh Bhaskar Yerrapragada , Nagendra Kundapura , Navaneeth Amin , Virupaksha Devaramane Pages 174-180
    Background
    Depression is a common and widespread mood disorder, which affects an emotional level that varies widely in its intensity. Biochemical parameter alterations have been observed in different depression types. In the present study, we examined acetylcholinesterase (AChE), paraoxonase 1 (PON1), and copper levels in moderately-depressed patients and healthy controls to ascertain whether the measurement of red blood cell (RBC) AChE, and plasma PON1 and copper could be used to evaluate moderate depression.
    Methods
    This case control study was performed in the Department of Biochemistry, Kasturba Medical College, Manipal Academy of Higher Education, Manipal, Karnataka, India. Patients who met ICD 10 diagnostic criteria were considered as cases. Goldberg’s General Health Questionnaire 28 (GHQ-28) was used to select controls. Four ml of blood was collected from 24 cases and 20 controls aged 35-70 years and used to determine RBC AChE, and plasma PON1 and copper levels.
    Results
    Red blood cell AChE, and plasma PON1 and copper levels were significantly greater in patients with moderate depression than in controls. Further, a receiver operating characteristic curve for validity of the biochemical parameters in plasma from patients with moderate depression indicated sensitivity and specificity above 85% for copper and PON1.
    Conclusions
    Red blood cell AChE, plasma PON1, and copper levels may have roles in the pathogenesis of depressive disorders
    Keywords: Acetylcholinesterase, Copper, Moderate Depression, PON1, Stress
  • Mostafa Shahmoradi , Peyman Faridifar , Reza Shapouri , Seyed Fazlollah Mousavi*, Mahnaz Ezzedin , Bahman Mirzaei Pages 181-188
     
    Background
    Among hospitalized patients, Staphylococcus aureus (S. aureus) infections pose a serious health threat. The present study investigated the frequency of biofilm forming genes among clinical isolates S. aureus and their susceptibility to antibiotics.
    Methods
    The clinical samples were analyzed via standard biochemical assays for identifying different bacterium, which was then confirmed using the multiplex colony PCR method. Those samples identified as S. aureus were examined for the presence of the cna, fnbA, fnbB and pvl genes. The antibiotic susceptibility of the S. aureus isolates was then tested.
    Results
    Using the standard biochemical assay approach, 54 S. aureus strains were identified. However, when using the multiplex PCR method 50 S. aureus strains were identified among the clinical samples. The in vitro biofilm formation assays determined 3 (6%) strains of S. aureus to be strong biofilm forming, 15 (30%) of the isolates were determined to be moderate biofilm forming and, 32 (64%) were determined to be weak biofilm forming. Among the isolated strains, the specific frequencies of the biofilm forming genes were determined to be 31 (62%) for cna, 35 (70%) for fnbA, 13 (26%) for fnbB and 1 (2%) for pvl. In 11 (22%) of the isolated strains fnbA, fnbB and cna genes were all present. All strains were determined to be penicillin, amoxicillin and clavulanic acid resistant.
    Conclusions
    Due to the increase of the antibiotic resistance in biofilm producing S. aureus strains, rapid identification of antibiotic resistance can help to eliminate the infection effectively
    Keywords: Biofilm, Multiplex colony PCR, Pertussis toxin, Spreading factors, Staphylococcus aureus
  • Hasan Mansouritorghabeh , Mojtaba Sankian , Abdol, Reza Varasteh , Malihe Moghadam , Bahman Khameneh , Farahzad JabbariAzad *, Reza FaridHosseini Pages 189-195
    Background
    The quality of extracts used in the skin prick test directly influences the interpretation of the test. Accordingly, the outcomes and effectiveness of immunotherapy for the management of IgE-mediated allergies depend on the quality of the extracts used. Excipients, which are pharmacologically inert ingredients, are intentionally added to the active ingredients. The aim of this study was to address optimum excipients for stability Platanus (P.) orientalis extract.
    Methods
    In this study the excipients examined were l-lysine (20 mM), l-cysteine (20 mM), albumin (0.5%), sorbitol (2%), sucrose (750 mM), trehalose (20 mM), D-mannitol (2% w/v), urea (100 mM) and Tween-20 (0.1%). Their effects on P. orientalis extract stability were analyzed using an inhibition enzyme linked immune assay at 37 ᵒC.
    Results
    A mixture of lysine (20 mM), trehalose (20 mM), and D-mannitol (2% w/v) conferred the greatest stability on the P. orientalis extract.
    Conclusions
    The P. orientalis extract stability was increased by a mixture of lysine (20 mM), trehalose (20 mM), and D-mannitol
    Keywords: Lysine, Mannitol, Platanus orientalis, Skin prick test, Trehalose
  • Ali Torkashvand, Fariborz Bahrami, Minoo Adib, Soheila Ajdary* Pages 196-203
    Background
    Lactic acid bacteria such as Lactococcus (L.) lactis are powerful tools that can function as live delivery vectors and heterologous protein expression hosts in development of novel vaccines. Pertussis toxin (PT) and filamentous hemagglutinin (FHA) are important virulence factors of Bordetella (B.) pertussis and constitute the major components of commercially available acellular pertussis (aP) vaccines. The purpose of the present study was to express F1S1 fusion protein, consisted of the N-terminal region of S1 subunit from PT and FHA type 1 immunodominant domain by L. lactis and to evaluate its immunogenicity.
    Methods
    The fusion gene composed of sequences encoding the F1S1 and the signal peptide of usp45 fragments (SECF1S1) was codon optimized for protein production in L. lactis and was synthesized and inserted in-frame inside pNZ8149 plasmid. The resulting pNZ8149-SECF1S1 construct was introduced by electroporation into L. lactis cells (LL-F1S1). BALB/c mice were subcutaneously immunized with LL-F1S1 or commercial DTaP vaccine. The immune responses were investigated.
    Results
    The LL-F1S1-immunized mice produced significant levels of specific IFN-g compared to their respective controls and DTaP-immunized mice. The F1S1- specific IgG antibody response was lower in LLF1S1- immunized mice while the IgG2a/IgG1 ratio was higher in this group compared to the DTaP-immunized mice. Moreover, anti-F1S1 IgA antibodies were only detected in the lung homogenates of the LL-F1S1- immunized mice, suggesting the induction of a mucosal immune response.
    Conclusions
    These results indicate the feasibility of expression of F1S1 fusion protein in L. lactis. This recombinant bacterium could induce mucosal and Th1-type systemic immune responses following subcutaneous administration.
    Keywords: Bordetella pertussis, FHA, Lactococcus lactis, Pertussis toxin
  • Mahmoud Amini, Abdolali Pakdaman, Shima Shapoori, Ghasem Mosayebi * Pages 204-209
    Background
    Acute cholecystitis is defined as gallbladder inflammation caused by obstruction of the cystic duct. The pro-inflammatory cytokine, high mobility group box-1 (HMGB1), has been found to hold critical roles in the pathogenesis of several different inflammatory diseases. This study aimed to determine the relationship between HMGB1 and acute cholecystitis, and examine the potential for this cytokine as a biomarker for clinical diagnosis.
    Methods
    The serum of 23 patients with severe acute cholecystitis, 45 patients with mild acute cholecystitis and 35 healthy subjects was collected and isolated from peripheral blood. The serum levels of HMGB1, CRP, amylase, lipase and the number of white blood cells were measured prior to the patient’s cholecystectomy and 48 hours following the procedure.
    Results
    A significant increase in the levels of HMGB1 were observed in both patient groups with mild or severe acute cholecystitis compared with normal group. ROC analysis determined a cut-off point of 2.34 for HMGB1 serum levels to discriminate between the normal group and acute cholecystitis patients with a sensitivity of 79.41% and a specificity of 54.3%. The area under the ROC curve was 0.71. Furthermore, a positive correlation was observed between CRP and HMGB1 levels and no significant difference in the levels of amylase and lipase was observed between groups.
    Conclusions
    These findings suggest a potential role for HMGB1 as an effective biomarker in improving the diagnostic accuracy of acute cholecystitis when used in conjunction with the standard diagnostic tests.
    Keywords: Cholecystitis, C- reactive protein, High mobility group box-1, Inflammation
  • Akbar Derakhshan, Javad Sadeghi Allah Abadi *, Jalil Tavakkol Afshari, Amin Reza Nikpoor, Ramin Daneshvar, Saeed Shokoohi Rad, Mohammad Reza Ansari Astaneh, Sadegh Ghafarian Pages 210-216
    Background
    Glaucoma is one of the main causes of irreversible blindness. The most common type of glaucoma is primary open angle glaucoma (POAG). TGF-β2, the main TGF-β isoform in the eye, is critical for extracellular matrix production and angiogenesis. Genetic studies have shown that TGF-β2 gene (TGFB2) polymorphisms affect its expression in the eye. The aim of this study was to investigate the presence of the TGFB2 rs991967 polymorphism in POAG, and the effect of this polymorphism on clinical characteristics in POAG patients.
    Methods
    This case-control study was conducted on 112 control participants and 112 POAG patients referred to Khatam-Al-Anbia Eye Hospital, Mashhad, Iran. The TGFB2 rs991967 polymorphism was genotyped by the PCR-restriction fragment length polymorphism (PCR-RFLP) method. The genotyping results and clinical findings were analyzed using SPSS version 16.
    Results
    The most common genotype was AA, observed in 54.5% of the patients (P < 0.0001, OR 12.2, CI 95% for OR: 5.25 to 28.31). Moreover, the highest and lowest frequencies of the mutant A allele were seen in the patient and control groups with percentages of 73 and 40%, respectively. This difference was significant (P < 0.0001, OR: 3.9, CI 95% for OR: 2.6 to 5.9). No significant association was seen between the frequencies of the TGFB2 rs991967 polymorphism genotypes and clinical characteristics in POAG patients.
    Conclusions
    The TGFB2 rs991967 polymorphism has a direct and significant association with POAG and significantly increases the risk of developing POAG.
    Keywords: PCR, Polymorphism, Primary Open Angle Glaucoma, TGF-beta2
  • Hoda Derakhshanian, Abolghassem Djazayery, Mohammad Hassan Javanbakht, Mohammad Reza Eshraghian, Abbas Mirshafiey, Mahnaz Zarei, Ehsan Alvandi, Ehsan Djalali, Mahmoud Djalali * Pages 217-222
    Background
    Diabetic nephropathy is one of the most important microvascular complications and a major cause of morbidity and mortality in diabetic patients. This study was designed to investigate the effect of vitamin D on the expression of three key genes involved in the development of diabetic nephropathy.
    Methods
    Twenty-four male Sprague–Dawley rats were randomly divided into three groups. The first group served as control and the other two groups received intraperitoneal injections of 45 mg/kg STZ to develop diabetes. The groups were treated for four weeks either with placebo or two vitamin D injections of 20,000 IU/kg. Serum glucose, insulin, and HbA1c levels, and AGE cellular receptor (RAGE), aldose reductase (AR) and glutamine: fructose-6-phosphate aminotransferase (GFAT) gene expression were assessed in kidney tissue at the end of the experiment.
    Results
    Vitamin D treatment resulted in a significant increase in insulin concentration, which could improve hyperglycaemia in diabetic rats. Serum HbA1c decreased slightly but insignificantly following the vitamin D injections. In addition, expression of GFAT, a key regulatory enzyme in the hexosamine pathway, was significantly reduced following vitamin D administration.
    Conclusions
    Vitamin D may reduce diabetic nephropathy not only by improving blood glucose and insulin levels, but also by modulating hexosamine pathways in kidney.
    Keywords: Diabetes Mellitus, Hexosamine pathway, Nephropathy, Vitamin D
  • Majid Hoseini *, Mehdi Sahmani, Farshad Foroughi, Yousef Khazaei Monfared, Mehdi Azad Pages 223-229
    Background
    Hypercoagulable states (HS) can result from several different inherited and acquired disease conditions that cause abnormalities in the genes, proteins and cellular factors involved in the coagulation cascade. Novel insight into the molecular mechanisms involved in the coagulation pathways can provide a framework to develop improved therapeutics to treat patients with coagulation disorders. Therefore, investigating the genetic abnormalities present in patients with coagulation disorders can offer critical insight into disease pathogenesis. Our study aimed to assess the promoter methylation patterns of the phosphatase and tensin homologue (PTEN) gene as a potential underlying factor involved in HS.
    Methods
    To measure the differences between the mRNA expression of PTEN in HS patients and healthy individuals we used qRT-PCR. Following bisulfite conversion, the promoter methylation status was analyzed using methylation specific PCR. The two-tailed student t-test was used to analyze the quantitative data. The data was considered statistically significant with a p value <0.05.
    Results
    Our findings reveal PTEN to be down-regulated by 30% in the blood samples of HS patients when compared to healthy controls. The MSP data showed the PTEN promoter region to be un-methylated in both patients and healthy individuals.
    Conclusions
    Since no differences in the methylation patterns of the PTEN gene was found between HS patients and controls, this suggests that DNA methylation of the PTEN promoter may not be a significant contributing epigenetic modification involved in the development HS. However, MSP may not be able to detect subtle changes in DNA methylation status. Thus, using an alternative high resolution technique may more accurately indicate differences in the PTEN promoter methylation status in HS patients.
    Keywords: Hyperquagulable State, Promoter methylation, PTEN