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FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ

Year 2006, Volume: 15 Issue: 2, 122 - 130, 01.06.2006

Abstract

İnsan periferal kan T-lenfositlerinin mitoza yönlendirilmesi ve kromozom analizlerinde birim hacimdeki büyüme ortamına eklenen fitohemaglütinin (FHE) miktarı bir laboratuardan diğerine göre önemli ölçüde değişmektedir. Bunun nedeni bilinmemekte ve literatürde de bu konuda yeterli bir bilgiye ulaşılamamaktadır. Alışılmış periferal kan kültürü (72 saat) ve kromozom hazırlama işlemi, yaşları 0-8 yıl arasında değişen Down Sendrom (DS)’lu hasta (N=30) ve sağlıklı kontrollerinden (N=24) alınan kan örneklerinin, aynı fakat konsantrasyonu gittikçe artan şekilde: 0.37 ml, 0.75 ml, 1.48 ml ve 2.21 ml FHE /100 ml’lik ortam’lara ekilmesi ile yapılmışlardır. FHE’nin dört farklı konsantrasyonunu içeren dört farklı ortamdan elde edilen her bir preparatta Mitotik indeks (Mİ)’ler, birey ve konsantrasyon başına 5000- 6000 uyarılmış lenfositler sayılarak yüzde (%) olarak belirlendi. Hem DS’lu, hem de kontrol gruplarının genel ortalamaları alındı. Bireyler arasında önemli farklılıklar görülmesine karşın, kültür ortamında FHE konsantrasyonunun artış yönüne doğru (%0.37, %0.75, %1.48, ve %2.21) lenfositlerdeki Mİ’lerin genel ortalamaları karşılıklı olarak; 30 DSlu’da %4.20±1.73, %4.57±1.31, %4.69±1.19 ve %4.19±1.11 iken 24 kontrolde %4.37±1.67, %5.06±1.60, %5.01±1.47 ve %4.98±1.43 olarak bulundu. Sonuçta, Biological İndustries (İsrail) ürünleri kullanıldığında, % 1-1.5 hacim/hacim FHE, hem DS’lu, hem de kontrollerin kan kültürleri için optimum (hem etkin hem de tutumlu) bulundu

References

  • Morimoto K, Kaneko T, Lijima K, Koizumi A. Proliferative kinetics and chromosome damage in trisomy 21 lymphocyte cultures exposed to γ- rays and bleomycin. Cancer Res 1984, 44:1499-1504.
  • Park E, Alberti J, Mehta P, Dalton A, Sersen E, et al. Partial impairment of immune func- tions in peripheral blood leukocytes from aged men with Down syndrome. Clin Immunol 2000, 95:62-69.
  • Anderson D, Jenkinson PC, Dewdney RS, Francis AJ, Godbert P et al. Chromosome aberration, mitogen induced blastogenesis and proliferative rate index in periferal lympho- cytes from 106 control individuals of the U.K. population. Mut Res 1988, 204:407-420.
  • Ross G, Dixon JB, Veevers A. Variation in the lymphocyte transformation assay slope analy- sis of cell dose-response curves. J Immunol Methods 1987, 98:189-193.
  • Tajima M, Fujinaga T, Okamoto Y, Otomo K, Koike T. Relatinship between mitogen receptor in peripheral blood lymphocytes and blasto- genic response to mitogen. Res Veter Sci 1990, 48:1-5.
  • Barta O, Barta V, Domermuth CH, Pierson FW. Optimum conditions for the turkey lym- phocyte transformation test. Avian Dis 1992a, 36:386-394.
  • Barta O, Barta V, Pierson FW. Optimum con- ditions for the chiken lymphocyte transforma- tion test. Avian Dis 1992b, 36:945-955.
  • İmamoglu N, Demirtas H, Donmez-Altuntas H, Hamurcu Z, İlten creases on metaphase chromosomes of Down syndrome lymphocytes, in concordance with the mitogen concentration in the culture me- dium. Cytometry Part B-Clin Cytometry 2005, 66B:36-39.
  • İmamoglu N, Demirtas H, İlten A. NOR Ex- pression increases in interphase lymphocytes of Down syndrome babies/children as AgNORs surface, according to the mitogen concentra- tion in the culture medium. Micron 2006, 37:129–133.
  • Nagel JE, Chopra RK, Chrest FJ, McCoy MT, Schneider EL, et al. Decreased proliferation, interleukin 2 synthesis and interleukin 2 recep- tor expression are accompanied by decreased mRNA expression in phytohemagglutinin- stimulated cells from elderly donors. J Clin Invest 1988, 81:1096-1102.
  • Farrant J, Clark JC, Lee H, Knight SC, O’Brien J. Condition for measuring DNA syn- thesis in PHA stimulated lymphocytes in 20 microliters hanging drops with various cell concentrations and period of culture. J İmmu- nol Meth 1980, 33:301-312.
  • İmamoğlu N, Demirtaş H, Dönmez H, Hamur- cu Z. Down sendromlu lenfositlerinde Kültür ortamına göre aktif NOR sayısı değişimi. VI. Ulusal Tıbbi Biyoloji Kongresi Özet Kitabı, ss: 173. 2-5 Kasım 2000, Pamukkale Üniversitesi, Denizli.

Effect of the Phytohemagglutinin Concentration on the Mitotik Indexes of Down Syndrome and Healthy Individuals’ Lymphocytes

Year 2006, Volume: 15 Issue: 2, 122 - 130, 01.06.2006

Abstract

The amount of phytohemagglutinin (PHA) amount added to the growing medium in unit volume of human peripheral blood T-lymphocytes induced mitosis and at chromosome analysis varies significantly from one laboratory to another. The reason for this variation is not known, and in literature adequate information on this subject is not available. Conventional peripheral blood culture (72h) and chromosome preparation procedure have been used except that blood samples from Down syndrome (DS) patients (N=30) and healthy controls (N=24), with the age range of 0-8 years, have been cultivated in the same but gradiently increasing PHA concentration containing: 0.37 ml, 0.75 ml, 1.48 ml and 2.21 ml of PHA / 100 ml of medium. Mitotic indexes (MIs) of each specimen obtained from four separate media with four different concentrations of PHA were determined as percentage (%) by screening from 5.000 to 6.000 stimulated lymphocytes per concentration per individual. Overall means MIs of both DS and control groups were taken. Although considerable interindividual variations were apparent, overall means of MIs in lymphocytes according to the gradient of PHA concentration (0.37%, 0.75%, 1.48%, and 2.21%) in culture medium were found as 4.20±1.73%, 4.57±1.31%, 4.69±1.19%, and 4.19±1.11% for 30 DS patients and 4.37±1.67%, 5.06±1.60%, 5.01±1.47%, and 4.98±1.43% for 24 healthy individuals, respectively. In conclusion, When the Biological Industries (Israel) products are used, 1-1.5 % volume/volume PHA was found optimum (both effective and economic) for the blood cultures of both DS patients and healthy controls

References

  • Morimoto K, Kaneko T, Lijima K, Koizumi A. Proliferative kinetics and chromosome damage in trisomy 21 lymphocyte cultures exposed to γ- rays and bleomycin. Cancer Res 1984, 44:1499-1504.
  • Park E, Alberti J, Mehta P, Dalton A, Sersen E, et al. Partial impairment of immune func- tions in peripheral blood leukocytes from aged men with Down syndrome. Clin Immunol 2000, 95:62-69.
  • Anderson D, Jenkinson PC, Dewdney RS, Francis AJ, Godbert P et al. Chromosome aberration, mitogen induced blastogenesis and proliferative rate index in periferal lympho- cytes from 106 control individuals of the U.K. population. Mut Res 1988, 204:407-420.
  • Ross G, Dixon JB, Veevers A. Variation in the lymphocyte transformation assay slope analy- sis of cell dose-response curves. J Immunol Methods 1987, 98:189-193.
  • Tajima M, Fujinaga T, Okamoto Y, Otomo K, Koike T. Relatinship between mitogen receptor in peripheral blood lymphocytes and blasto- genic response to mitogen. Res Veter Sci 1990, 48:1-5.
  • Barta O, Barta V, Domermuth CH, Pierson FW. Optimum conditions for the turkey lym- phocyte transformation test. Avian Dis 1992a, 36:386-394.
  • Barta O, Barta V, Pierson FW. Optimum con- ditions for the chiken lymphocyte transforma- tion test. Avian Dis 1992b, 36:945-955.
  • İmamoglu N, Demirtas H, Donmez-Altuntas H, Hamurcu Z, İlten creases on metaphase chromosomes of Down syndrome lymphocytes, in concordance with the mitogen concentration in the culture me- dium. Cytometry Part B-Clin Cytometry 2005, 66B:36-39.
  • İmamoglu N, Demirtas H, İlten A. NOR Ex- pression increases in interphase lymphocytes of Down syndrome babies/children as AgNORs surface, according to the mitogen concentra- tion in the culture medium. Micron 2006, 37:129–133.
  • Nagel JE, Chopra RK, Chrest FJ, McCoy MT, Schneider EL, et al. Decreased proliferation, interleukin 2 synthesis and interleukin 2 recep- tor expression are accompanied by decreased mRNA expression in phytohemagglutinin- stimulated cells from elderly donors. J Clin Invest 1988, 81:1096-1102.
  • Farrant J, Clark JC, Lee H, Knight SC, O’Brien J. Condition for measuring DNA syn- thesis in PHA stimulated lymphocytes in 20 microliters hanging drops with various cell concentrations and period of culture. J İmmu- nol Meth 1980, 33:301-312.
  • İmamoğlu N, Demirtaş H, Dönmez H, Hamur- cu Z. Down sendromlu lenfositlerinde Kültür ortamına göre aktif NOR sayısı değişimi. VI. Ulusal Tıbbi Biyoloji Kongresi Özet Kitabı, ss: 173. 2-5 Kasım 2000, Pamukkale Üniversitesi, Denizli.
There are 12 citations in total.

Details

Other ID JA47CU53RE
Journal Section Research Article
Authors

Nalan İmamoğlu This is me

Halil Demirtaş This is me

Publication Date June 1, 2006
Submission Date June 1, 2006
Published in Issue Year 2006 Volume: 15 Issue: 2

Cite

APA İmamoğlu, N., & Demirtaş, H. (2006). FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ. Sağlık Bilimleri Dergisi, 15(2), 122-130.
AMA İmamoğlu N, Demirtaş H. FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ. JHS. June 2006;15(2):122-130.
Chicago İmamoğlu, Nalan, and Halil Demirtaş. “FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ”. Sağlık Bilimleri Dergisi 15, no. 2 (June 2006): 122-30.
EndNote İmamoğlu N, Demirtaş H (June 1, 2006) FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ. Sağlık Bilimleri Dergisi 15 2 122–130.
IEEE N. İmamoğlu and H. Demirtaş, “FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ”, JHS, vol. 15, no. 2, pp. 122–130, 2006.
ISNAD İmamoğlu, Nalan - Demirtaş, Halil. “FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ”. Sağlık Bilimleri Dergisi 15/2 (June 2006), 122-130.
JAMA İmamoğlu N, Demirtaş H. FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ. JHS. 2006;15:122–130.
MLA İmamoğlu, Nalan and Halil Demirtaş. “FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ”. Sağlık Bilimleri Dergisi, vol. 15, no. 2, 2006, pp. 122-30.
Vancouver İmamoğlu N, Demirtaş H. FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ. JHS. 2006;15(2):122-30.