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Golestani R, Moazzeni S. M, Puorfathollah A.a, Hamidpour L, Sharafi M, Attarchi Z, Hadji-Beygi B, Tavasoli F, Esmaili J, Reyvandi S,
Volume 3, Issue 3 (Autumn 2006)
Abstract

Abstract

Background and Objectives

Providing fetal calf serum (FCS) alternatives as cell culture supplements is an important field of research to compensate for the FCS supply shortage.This study focused on preparation of fetal calf serum alternatives and their effects on growth and secretion of hybridoma cell lines.

Materials and Methods

Outdated human platelet units undergo extraction for its growth factors to be obtained. Human AB blood group plasma was also converted to serum and its growth effect was compared to FCS, hypoxanthine-thymidine (HT) and RPMI1640 as cell culture media and supplement. Cell growth indices were preliminary counting of cells , confluency as surface area of plates filled with cells, and titration of monoclonal anti-A and anti-B blood group antibodies collected from cultured mouse hybridoma cells. Statistical analysis including one sample t-test, logarithmic multiple regression curve fit, and factor analysis was done by SPSS v12 software.

Results

The four nutritional supplements of (1) human serum AB (AB), (2) human platelet extract (PLT), (3) equal mixture of AB & PLT (ABP), and (4) fetal calf serum as cell culture were examined on mouse hybridoma anti-A and anti-B monoclonal antibody producer cell lines for cell growth indices and compared with the same indices on RPMI1640 media. The growth-stimulating effects in descending order of values were (1) ABP5% , (2) FCS10% , (3) ABP10% , (4) AB5% , (5) AB10% , (6) PLT5% , (7) ABP20% , (8) PLT10% , (9) PLT20%, and (10)HT but AB20% inhibited growth of mentioned hybridoma cell lines. The titer of anti-A and anti-B monoclonal antibodies produced by cultured hybridoma on 5 and 10 percent concentration of AB , PLT and ABP compared to FCS5-10% at descending order were (1) PLT5%, (2) PLT10% , ABP5% , ABP10% , AB10%, and (3) AB5%.

Conclusions

In general FCS had the following effects on curves of cell growth: (1) the highest increase on slope of multiplication (ascending) phase, (2) the highest increase on slope of death (descending) phase, and (3) the lowest duration of stationary phase. Then, FCS can be appropriate for growth of cells at initial low cell count. Human serum AB, human platelet extract, and equal mixture of both at optimum concentrations (these supplements at high concentrations killed cells) compared to FCS showed (1) decreased slope of multiplication phase, (2) decreased slope of death phase, and (3) increased duration of stationary phase. Thus, AB and PLT may be suitable for continuous cell culture systems in which cell survival during longer times is required. Factor analysis was introduced as a model to evaluate kinetics of cell growth at different supplements.

Key words : Human serum, Cell culture, Growth kinetic, Blood groups, Factor analysis


Ladan Hosseini Gohari, Isa Noormohammadi, Ali Akbar Sharafi Tafresi Moghadam, Leila Mostaan, Aneti Drousiotou,
Volume 3, Issue 4 (Winter 2007)
Abstract

 

  Abstract

 Background and Objectives

 For a successful prevention program, globin chain synthesis, as a complementary test beside DNA analysis, is necessary. So, it is important that each laboratory establishes its own reference range for the classification of thalassemia syndromes by globin chain synthesis. Globin chain synthesis is a relatively complex test introduced in the study of thalassemia syndromes as a reference method. The technique is also useful for variant chain identification.This study aims to stablish the method of globin chain synthesis to determine a / b chain ratio in healthy individuals.

 

 Materials and Methods

 In this study globin chain analysis was performed on 30 healthy laboratory personnels with normal HbA2 and normal hematological indices. In this method a reticulocyte-rich sample is incubated with a mixture of amino acids, one of which (leucine) radioactively labelled. After washing the excess radioactivity and precipitating the globin, different chains are separated by cation exchange chromatography.

 

 Results

 The mean a / b ratio was 1.045 ± 0.12 (mean ± 1SD) in healthy subjects. Our findings were in agreement with those of the other investigators in the world.

 

 Conclusions

 In any screening program, diagnostic problems will arise that can not be solved without biosynthetic studies. The Clegg and Weatherall method has been proved to be very reliable and reproducible, but time-consuming (requiring four days). New methods like reverse phase HPLC are now available for chain separation. Therefore, according to the procedures each laboratory should determine its own reference range.

  

 Key words : a globin, b globin, Hemoglobin, Normal range, Ion exchange chromatography

 


A. Talebian, A.r. Shafaei, M. Sharafi, S. Rivandi, L. Jalili, T. Fattaheian,
Volume 7, Issue 1 (Spring 2010)
Abstract

 

 

  Abstract

 Background and Objectives

 ABO blood group system antibodies agglutinate red blood cell suspension in physiologic serum directly without any reaction booster. In the present study the potency of IBRF-manufactured anti-A and anti-B blood grouping reagents by the hemagglutination method in test tube was reassessed against the new WHO international minimum potency standards.

 

 Materials and Methods

 In this experimental study, starting concentrations defined in WHO standards were used in titration. Doubled dilution series of WHO standards (from starting concentrations) and IBRF blood grouping reagents (from neat) were prepared by using buffered saline containing 2% BSA as diluent. One volume of each starting concentration together with one volume of prepared dilutions were mixed with one volume of a 2% suspension of A1, A2, A2B, and B cells in glass test tubes, respectively. After appropriate incubation and centrifugation of the tests according to specified criteria, the reactions were graded macroscopically.

 

  Results

 The results showed that the IBRF anti-A and anti-B blood grouping reagents comply with the minimum WHO standard dilution. Consequently, IBRF anti-A and anti-B blood grouping reagents were shown to be safe for screening and diagnostic purposes.

 

 Conclusions

 The quality of blood grouping reagents is clearly an important factor for safe blood transfusion. Routine titration tests are not reliable methods for evaluation of those reagents. Fortunately, this problem would be solved using the above standards. This recommended method is provided to help assist manufacturers in pursuing new product license applications and making amendments in existing ones. 

  

 Key words : Reagents, Agglutination, Standards

 


A. Safaei, Dr. F. Zaker, H. Sharafi, Dr. M. Hashemi, Dr. P. Yaghmaei, M. Abdolahzadeh,
Volume 8, Issue 3 (Autumn 2011)
Abstract

  Abstract

 Background and Objectives

 Many studies have demonstrated that polymorphisms of NQO1 including C465T and C609T are associated with increased risk of acute myeloid leukemia(AML). Our aims are to assess incidence of these polymorphisms in Tehran patients and study the influence of low activity of NQO1 in AML.

 

 Materials and Methods

 In this case-control study, we used PCR and RFLP analyses to study the prevalence of C609T NQO1in 140 patients, and C465T NQO1 in 124 patients there was also a control group of 80 being age-sex matched. We calculated odd ratio with SPSS 16 to examine if these polymorphisms are associated with AML.

  

 Results

 No significant association between the two common polymorphisms of NQO1 and risk of AML was observed. C609T odd ratio for TT genotype versus CC was obtained to be 0.91 (CI 95% = 0.51-1.63) and for CT versus CC it was 1.06 (CI 95% = 0.57-1.95). C465T odd ratio for TT genotype versus CC was calculated to be 0.22 (CI 95% = 0.009-5.56) and for CT versus CC it came out to be 3.01(CI 95% = 0.63-14.32).

  

 Conclusions

 Our findings suggest that the NQO1 C609T and C465T gene variants do not have a major influence on the susceptibility to adult AML.



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