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Articles by P. Ghadam
Total Records ( 9 ) for P. Ghadam
  S. Sadrai , P. Ghadam , R. Sharifian , E. Nematipour , Z. Kianmehr and Sh. Shahriari
  In this study, we investigated pharmacokinetic of warfarin in sensitive and control patients by HPLC. The analysis performed by HPLC consisted of a column (Perfectsil Target, 5 μm, 125x4.0 mm) and an isocratic mobile phase of methanol: acetonitrile: phosphate buffer (pH: 3.5) (7:55:38, v/v), flow rate: 1 mL min-1 and UV detection at 270 nm. The assay was linear in warfarin concentration ranges of 0.1-10 μg mL-1 (r2 = 0.9975) and with Relative Standard Deviation (RSD) of <8% for inter-day and <6% for intra-day. The mean warfarin blood concentration of 54 patients was 1.2 ± 0.6 μg mL-1.
  P. Ghadam , F. Sadeghian , R. Sharifian , S. Sadrai , B. Kazemi and E. Nematipour
  The vitamin K epoxide reductase subunit 1 (VKORC1) has been identified recently. It is a component of the enzyme vitamin K epoxide reductase that is the therapeutic target site of warfarin. In order to investigate the relationship between VKORC1 gene and warfarin dose response, we studied this gene in an Iranian warfarin resistant patient who receive more than 100 mg warfarin per day. The results showed that although warfarin concentration in his plasma was extremely higher than therapeutic level (22.8 mg L-1) but no mutation(s) found in the exons of VKORC1 gene. Other genes may be contributed in resistance to warfarin in this patient.
  P. Ghadam , A. Abadi , A. Asadzadeh , N. Safari , A. Shabani and M. Sharifian
  In this study, we found that allelic polymorphism in Hp gene acts as a major determinant of susceptibility for the development of diabetic cardiovascular complications. We studied this gene in 122 Iranian diabetic cardiovascular patients. The results showed that distribution of the Hp phenotypes were found to be in Hardy-Weinberg equilibrium. By using the chi square test we determined the association between Hp allele and diabetic cardiovascular diseases (χ2 = 52.98 p<0.001) and this finding was independent of gender (χ2 = 0.39 p>0.05). The chance of having allele two in diabetic patients with cardiovascular disease was 22.31 (95% CI, 6.18 to 80.5) times more than those with allele one (p<0.0001).
  S. Sadrai , P. Ghadam , R. Sharifian and F. Sadeghian
  A simple and rapid HPLC method with UV detecting system has been used in determination of warfarin level in plasma of Iranian patients who received different doses of this drug. Six resistance (10-70 mg day-1) and 5 sensitive patients (0.5-2.5 mg day-1) were selected for this study. Range of warfarin level in plasma was between 0.93 and 22.8. After determination of warfarin level in warfarin sensitive and especially, warfarin resistance patients, we are going to find a relationship between this level and pharmacokinetic or pharmacogenetic factors. In the separate study which was done in our laboratory on the gene that is possibly responsible for warfarin resistance we did not find any mutation in our patient with high warfarin concentration in their blood.
  M. Darvish , S.A. Ebrahimi and P. Ghadam
  Phenylketonuria (PKU) and Maple Syrup Urine Disease (MSUD) are two inborn metabolic diseases which are carried by autosomal recessive genes in man. These genetic errors result in accumulation of phenylalanine (in PKU) or valine, leucine and isoluecin (in MSUD). At high concentrations, amongst other problems, these amino acids cause mental retardation. However if detected early after birth, using special diets and other forms of therapy, mental abnormalities can be prevented. As a result in many countries screening of infants for MSUD and PKU, by measuring plasma amino acids has become a routine neonatal test. Capillary Electrophoresis (CE) assays have a number of advantages over the traditional chromatography techniques (such as GC or HPLC). These include low cost, high speed of analysis and high resolution. These characteristics, make CE an ideal method for the screening of inborn errors of metabolism. We developed a CE assay based on pre-column derivatisation of amino acids with phenylisothiocyanate. This conjugate has strong absorbance at 254 nm. CE was carried out using a Spectraphoresis 1000 instrument, fitted with 40 cm of a 25 μm capillary, at 17°C. A running voltage of 18KV was used to separate the amino acid mixture in an electrophoretic buffer containing 45 mM imidazole, 6 mM borate and 208 mM SDS, fixed at pH 9 with 2-N-morpholino ethane sulfonic acid. The assay was calibrated using various concentrations of amino acid standards. LOD, LOQ, recovery, inter-day and intra-day variations of the assay were determined. Also, levels of the 4 amino acids in normal and abnormal plasma were determined and compared with HPLC.
  P. Ghadam , N. Shariatian , M.A. Amoozegar , A. Rabbani and S.H. Shahriari
  Histone-Like Proteins (HLPs) in bacteria are small basic proteins that contribute to the control of gene expression, recombination, DNA replication and compressing the bacterial DNA in the nucleoid. Among the HLPs, HU protein as a dimer plays an important role in binding to DNA and bending it. In this study, we showed that a 9.5-10 kDa protein with the same electrophoretic mobility as HU exists in Halobacillus karajensis which is a novel gram positive moderate halophile bacterium that was recently isolated from surface saline soil of the Karaj Region, Iran. The genes encoding HU protein were also assayed during this study by Polymerase Chain Reaction.
  P. Ghadam , S. Gharavi , F. Yarian , M.R. Soudi , B. Kazemi and M. Bandehpour
  Among some Bacillus species, a protein highly homologous to HU, classified HB and coded by hbs gene. According to the recent studies, the sequence of hbs gene just in one strain of Bacillus subtilis exists in gene bank (ATCC 23857). In this study, DNA from Bacillus subtilis ATCC 6633 was extracted and investigated by PCR. The PCR product was sequenced and shown to differ in just one nucleotide from B. subtilis ATCC 23857. Hence, it was chosen as reference and for the first time, used for non-radioactive labeled probe preparation. The PCR product in Bacillus subtilis with ATCC 6633 was labeled using non-radioactive DIG-labeled nucleotides and conditions of probe preparation and hybridization were optimized and checked it by Southern blotting.
  P. Ghadam , R. Sharifian , Z. Jomeh Farsangi , Z. Kianmehr and M. Lak
  For the first time in this study, the pharmacogenetic effects of CYP2C9 polymorphism on warfarin sensitivity in some Iranian patients who are on warfarin treatment were shown. The study group consisted of clinically sensitive patients(21 patients) and the control group (37 adult patients). For detection of CYP2C9*2 and CYP2C9*3 variants, a protocol based on restriction fragment length polymorphism based polymerase chain reaction with Eco47I and KpnI was used. In clinically sensitive patients about 81% and in normal response patients about 24.3% carried variant genotypes.
  Z. Kianmehr , P. Ghadam , S. Sadrai , B. Kazemi and R.A. Sharifian
  VKORC1 is a component of the enzyme that is the therapeutic target site of warfarin. In order to investigate the relationship between VKORC1 gene and warfarin dose response, we studied this gene in 22 clinically sensitive patients and 36 clinically normal patients as control group that in previous studies their blood warfarin levels were determined by HPLC and genotyped for CYP2C9. In the majority of these patients, the results had shown that the clinical phenotype was according to the CYP2C9 genotype, but there were sensitive patients with normal CYP2C9 genotype. So in this study for investigation of another reason of sensitivity to warfarin in these patients, VKORC1 was chosen. In order to determine the impact of VKORC1 1173C>T and 3730G>A polymorphisms in warfarin dose variability, a protocol of RFLP based PCR with HinfI and SsiI was used. Polymorphisms of VKORC1 were effective factors in sensitivity to warfarin in the majority of them. However, in some of sensitive patients, there was inconformity of clinical phenotypes with CYP2C9 and VKORC1 genotypes. In conclusion, our findings suggest that CYP2C9 and VKORC1 are not the only effective factors in sensitivity to warfarin and more studies are necessary for investigation of sensitivity reasons in these patients.
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