Peripheral blood analysis

De Ferrari M, Artuso M, Bonassi S, et al. 1991. Cytogenic biomonitoring of an Italian population exposed to pesticides Chromosome aberration and sister-chromatid exchange analysis in peripheral blood lymphocytes. Mutat Res 260 105-113. 

Byrn RA, Kiesshng AA (1998) Analysis of human immunodeficiency virus in semen indications of a genetically distinct virus reservoir. J Reprod Immunol 41(1-2) 161-176 Carr JM, Hocking H, Li P, Burrell CJ (1999) Rapid and efficient celL-to-ceU transmission of human immunodeficiency vims infection from monocyte-derived macrophages to peripheral blood lymphocytes. Virology 265(2) 319-329... 

Wetzel MA, Steele AD, Eisenstein TK, Adler MW, Henderson EE, Rogers TJ (2000) Mu-opioid induction of monocyte chemoattractant protein-1, RANTES, and IFN-gamma-inducible protein-10 expression in human peripheral blood mononuclear cells. J Immunol 165 6519-6524 Widmer U, Manogue KR, Cerami A, Sherry B (1993) Genomic cloning and promoter analysis of macrophage inflammatory protein (MIP)-2, MIP-1 alpha, and MIP-1 beta, members of the chemokine superfamily of proinflammatory cytokines. J Immunol 150 4996-5012 Ye RD (2001) Regulation of nuclear factor kappaB activation by G-protein-coupled receptors. [Review] [136 refs]. J Leukoc Biol 70 839-848... 

Anonymous. Allogeneic peripheral blood stem-cell compared with bone marrow transplantation in the management of hematologic malignancies An individual patient data meta-analysis of nine randomized trials. J Clin Oncol 2005 23 5074-5087. 

Another peculiarity of the study is that the use of a biological system has allowed the authors to hypothesize a possible mechanism of action of the leachate as a mixture, hypothesis that could have been drafted on the basis of the only knowledge derived by chemical analysis. Researchers suggest that leachate inhibits cell proliferation at low doses probably inducing a reversible cell cycle arrest that becomes irreversible at high doses, probably due to leachate-induced oxidative stress. This activity is mainly due to the chemical compounds extracted in the aqueous phase. Similar effects were noticed by previous investigations on other human cells (human peripheral blood lymphocytes and a human breast cancer cell line, MCF-7) [31, 32], supporting the hypothesis that cells that survive the initial insult from leachate constituents maintains the potential to proliferate until the effects on cell metabolism lead to death. 

Weaver, J.L. et al., Serial phenotypic analysis of mouse peripheral blood leukocytes, Toxicol. Mech. Methods, 12, 95, 2002. 

Metaphase Analysis. Metaphase analysis can be performed in any tissue with actively dividing cells, but bone marrow is the tissue most often examined. Cells are treated with a test compound and are arrested in metaphase by the administration of colcemid or colchicine at various sampling times after treatment. Preparations are examined for structural chromosome damage. Because the bone marrow has a good blood supply, the cells should be exposed to the test compound or its metabolites in the peripheral blood supply. Additionally, these cells are sensitive to S-dependent and S-independent mutagens (Topham et al., 1983). 

Evans, H.J. and O Riordan, M.L. (1975). Human peripheral blood lymphocytes for the analysis of chromosome aberrations in mutagen tests. Mutation Res. 31 135-148. 

Assay for human immunodeficiency virus type 1 (HIV-1) proviral DNA in peripheral blood monuclear cells can be performed by PCR followed by detection of PCR products by electrochemiluminescence-labeled oligonucleotide probe [Tris-bipyridine ruthenium (II) complex]. Since one of the PCR primers is biotin-labeled at the 5 end, facile capture of the PCR product-probe complex can be accomplished on streptavidin-conjugated magnetic particles, prior to analysis in an electrochemiluminescence analyzer (S3). 

Detection of chromosome fragments. Individuals with Turner s syndrome are, in some cases, mosaic for a portion of or for the entire Y chromosome (46,XY/45,X). Since such individuals may be at increased risk for gonadal tumors, Southern blot or PCR analysis has been used to detect the presence of Y chromosome segments in studies of DNA from peripheral blood samples. Similarly, the fetal sex as well as the presence of some aneuploid states (e.g., trisomy 18) can be determined by analysis of DNA from chorionic villi or amniotic fluid cells. 

Lombardi, V.R.M., Amado, L., Femandez-Novoa, L., Ftcheverrfa, 1., Seoane, S., Cacabelos, R. (2003) Flow cytometry analysis of CD28-/CD8-I- suppresor cell precursor and CD45RO-I-/ CD4-I- memory T lymphocytes in the peripheral blood of Alzheimer s disease patients. In New trends in Alzheimer- and Parkinson-related disorders, Hanin, 1., Fisher, A., Cacabelos, R. (eds.), Monduzzi Fditore, Bologna, pp. 57-61. 

Studies performed at RCRM have shown that hematopoietic and immune systems reconstitution after irradiation depends greatly on the functional abilities of the stem cells. Subset analysis and expression of CD34-i- antigens on bone marrow and peripheral blood cells were studied in Chernobyl accident clean-up workers including patients with leukemia and myelodysplasia and patients exposed to the natural levels of irradiation (table 2). 

Theunissen, K. Verfaillie, C.M. (2005) A multifactorial analysis of umbilical cord blood, adult bone marrow and mobilized peripheral blood progenitors using the improved ML IC assay. Experimental Hematology, 33,165-172. 

Immunotoxicity. There are currently no data on the effects of 2-hexanone on the human immune system via any route of exposure. Animal data included an inhalation study in which there was a 40% decrease in peripheral white blood cells in rats exposed to 2-hexanone (Katz et al. 1980). In addition, 2,5-hexanedione, a metabolite of 2-hexanone, was shown to adversely affect lymphoid organs of the immune system in rats and to cause impairment of immunity in mice (Upreti and Shanker 1987). Immunological assessments, including analysis of peripheral blood components and effects on lymphoid tissue, conducted as part of intermediate-or chronic-duration studies and skin sensitization tests would be useful in developing a dose-response relationship and assessing the potential risk to chronically exposed persons in the vicinity of hazardous waste sites or to exposed workers. 

Fig. 9. Flow analysis of apoptotic human peripheral blood lymphocytes using direct TUNEL assay. Human peripheral blood lymphocytes (1 x 10 ) treated (A) without or (B) with dexamethasone (0.1 ijlM) for 16 h were transferred to a 15-ml tube. Paraformaldehyde (2%) was added to cells with shaking and incubated for 10-15 min in ice with occasional shaking. Cells were washed with PBS with 1% BSA and 3-4 ml cold acetone was then added to cells. After 2-3 min incubation on ice with occasional shaking, cells were washed twice and TUNEL reaction mixture including enzyme TdT and fluorescein-labeled anti-dUTP antibody was added to cells (H). For the negative control group, only label solution without TdT was added to cells ( ). Cells without any addition of reaction or label solution were used for assessment of the autofluorescence ( ). The cell mixture was incubated 1 h at 37°C in the dark. The result of the apoptosis after flow analysis was expressed as a histogram using software CellQuest (Becton Dickinson). In Fig. 9A Ml (nonapoptotic cells), 86% M2 (apoptotic cells), 14%. In Fig. 9B Ml (nonapoptotic cells), 78% M2 (apoptotic cells), 22% (our unpublished data).

The specimen will be the basis for the analytic analysis. Is it RNA or DNA What is the origin of the tissue Amniocentesis Was it a spontaneous product of conception Were anatomic pathology slides or tissue blocks prepared Are cell lines involved Are these primary or immortalized Was a chorionic villus sampling procedure done Is the sample properly collected peripheral blood The answers to each of these questions should be noted, and considered part of the validation of a useful nucleic acid extraction method. A molecular diagnostics laboratory should adhere to the highest standards in providing services, and prior validation of applicable nucleic acid extraction procedures is a must to ensure high-quality service. 

Elias Z, Mur JM, Pierre F, et al. 1989. Chromosome aberrations in peripheral blood lymphocytes of welders and characterization of their exposure by biological samples analysis. J Occup Med 31 477-483. 

Fomenko, V.N. Katosova, L.D. (1973) The results of cytogenetic analysis of the peripheral blood in women workers in contact with chloroprene latex. In Kazan, Nauchnye Trudy Akusherkso-Ginekologich-eskaya Profpatologiya, pp. 33-37 (in Russian)... 

Darroudi, F. Natarajan, A.T. (1983) Cytogenetic analysis of human peripheral blood lymphocytes in vitro treated with resorcinol. Mutat. Res.. 124. 179-189... 

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