Characterization of a rat lymphocyte culture system for assessing sister chromatid exchange. II. Effects of 5-bromodeoxyuridine concentration, number of white blood cells in the inoculum, and inoculum volume

1982 ◽  
Vol 4 (5) ◽  
pp. 585-594 ◽  
Author(s):  
A. D. Kligerman ◽  
J. L. Wilmer ◽  
G. L. Erexson
Keyword(s):  
Sister Chromatid ◽  
Blood Cells ◽  
Sister Chromatid Exchange ◽  
Culture System ◽  
White Blood Cells ◽  
Lymphocyte Culture ◽  
Inoculum Volume

The Characterization of DYRK3 mRNA and the Utility of DYRK3 Inhibitors in a Carboplatin/Radiation Mouse Model of Anemia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2202-2202
Author(s):  
Connie L. Erickson-Miller ◽  
Ying Homan ◽  
Matthew Chomo ◽  
Elizabeth I. Valoret ◽  
Louis Elefante ◽  
...  
Keyword(s):  
Single Dose ◽  
Mouse Model ◽  
Blood Cells ◽  
Radiation Treatment ◽  
White Blood Cells ◽  
Negative Regulator ◽  
Mrna Levels ◽  
Erythroid Progenitors ◽  
Low Levels

Abstract DYRK3, a member of the dual-specificity tyrosine phosphorylation-regulated kinase family, is expressed at low levels in erythroid progenitors and is implicated as a negative regulator of erythropoiesis. An appropriate animal model of anemia was sought to investigate the potential utility of DYRK3 inhibitors as a therapy for the treatment of anemia. Treatment of mice with sub-lethal irradiation followed by a single dose of the chemotherapeutic, nucleoside analog carboplatin, is well known to induce severe anemia. In the first instance, an analysis of DYRK3 mRNA levels was performed to ascertain whether the anemia so induced results in increased DYRK3 transcription. In this carboplatin/radiation model, mice were exposed to sub-lethal gamma irradiation (500 rads) followed by a single dose of carboplatin (60 mg/kg). This treatment induced an approximately 50% decrease in hemoglobin with concomitant drops in other erythroid parameters (hematocrit and RBC) by days 15–17. The following parameters were examined in five animals per day: peripheral blood counts, marrow cell count, plasma Epo levels, marrow Ter119+/CD71+ expression and marrow DYRK3 mRNA. As expected, the hemoglobin, hematocrit and red blood cells decreased gradually to nadir on day 15. White blood cells decreased to very low levels within 2 days of carboplatin/radiation treatment and remained suppressed for 11 days. Platelets decreased to nadir at day 7, where they remain until day 10. Plasma Epo levels were low and abruptly increased at day 3–4. The absolute number of Ter119+/CD71+ cells immediately dropped from normal levels at day 1 and then increased at day 6 and then fluctuated between a 30- to 60- fold enhancement from day 8 through 21 when the study was complete. DYRK3 mRNA, as measured by quantitative PCR (Taqman), increased approximately 10-fold at day 7 and remained in that range until day 21. The number of erythroid progenitors measured by flow cytometry,Ter119+/CD71+ cells, and the level of DYRK3 mRNA remained elevated until the end of the experiment at day 21, at which point the hemoglobin had recovered to near normal levels. GSK626616, a potent, low molecular weight inhibitor of DYRK3 kinase activity (IC50 = 0.7 nM), was dosed once daily, i.p., for 17 days in this model. At day 15, the GSK626616-treated mice (0.03 mg/kg) demonstrated a statistically significant increase in hemoglobin, hematocrit, red blood cell and platelet counts compared to anemic animals treated with vehicle alone. In contrast to its effects in anemic mice, this compound demonstrated no increases in any blood parameters in normal mice over a similar timeframe and dosage regimen. This expected behavior is hypothesized to be due to the low level of DYRK3 mRNA in normal, non-anemic mice. The characterization of this carboplatin/radiation mouse model demonstrates that as the hemoglobin decreased, plasma Epo increased at day 3–4, followed by the increase in Ter119+/CD71+ cells at day 6. Following this surge in erythropoiesis, an increase in DYRK3 mRNA expression naturally follows. The subsequent improved erythropoiesis in animals treated with a DYRK3 inhibitor in this model, suggests that DYRK3 kinase mRNA levels could have utility as a biomarker in the identification of an anemic patient population that then may be a candidate for treatment with a DYRK3 inhibitor.

scholarly journals Reliable assessment of bone marrow and bone marrow concentrates using automated hematology analyzer

2019 ◽  
Vol 14 (7) ◽  
pp. 639-646 ◽  
Author(s):  
Venkata P Mantripragada ◽  
Nicolas S Piuzzi ◽  
Jaiben George ◽  
Wesley Bova ◽  
Mitchell Ng ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Red Blood Cells ◽  
Blood Cells ◽  
White Blood Cells ◽  
Limiting Factor ◽  
Cell Therapies ◽  
Reliable Assessment ◽  
Hematology Analyzer ◽  
Automated Hematology Analyzer

Aim: A limiting factor in advancement of bone marrow based cell therapies is the lack of characterization of cell products delivered to patients. Methods: Using an automated hematology analyzer that can be implemented in clinical setting, the composition of bone marrow aspirates (n = 17 patients) and bone marrow concentrates (n = 12 patients) were assessed. ICC estimates were calculated for measuring reliability. Results: Bone marrow aspirates assessment resulted in excellent reliability for determining white blood cells (ICC – 0.96; 95% CI: 0.92–0.99), red blood cells (ICC – 0.9; 95% CI: 0.77–0.96), platelets (ICC – 0.93; 95% CI: 0.85–0.97) composition. Bone marrow concentrate assessment resulted in excellent reliability for determining white blood cells (ICC – 0.97; 95% CI: 0.93–0.99), platelets (ICC – 0.95; 95% CI: 0.89–0.99) and moderate reliability for red blood cells (ICC – 0.66; 95% CI: 0.36–0.87) composition. Conclusion: Modern automated hematology analyzers could assist to better characterize the cell therapy products to provide reliable and consistent outcomes.

Characterization of novel CAPN3 isoforms in white blood cells: an alternative approach for limb-girdle muscular dystrophy 2A diagnosis

Neurogenetics ◽  
2008 ◽  
Vol 9 (3) ◽  
pp. 173-182 ◽  
Author(s):  
L. Blázquez ◽  
M. Azpitarte ◽  
A. Sáenz ◽  
M. Goicoechea ◽  
D. Otaegui ◽  
...  
Keyword(s):  
Muscular Dystrophy ◽  
Blood Cells ◽  
White Blood Cells ◽  
Limb Girdle Muscular Dystrophy ◽  
Alternative Approach

scholarly journals DEMONSTRATION AND CHARACTERIZATION OF TWO DISTINCT HUMAN LEUKOCYTIC PYROGENS

1974 ◽  
Vol 139 (6) ◽  
pp. 1369-1381 ◽  
Author(s):  
Charles A. Dinarello ◽  
Nathan P. Goldin ◽  
Sheldon M. Wolff
Keyword(s):  
Isoelectric Point ◽  
Blood Cells ◽  
White Blood Cells ◽  
Gel Filtration ◽  
Human Monocytes ◽  
First Time ◽  
Human White Blood Cells ◽  
Pyrogenic Activity

Human monocytes and neutrophils were separated from buffy coats of blood obtained from normal donors. Following incubation with heat-killed staphylococci, monocyte preparations contained 20 times more pyrogenic activity in the supernatant media than did supernates from an equal number of neutrophils. During purification of these pyrogens it was discovered that these cell preparations each produced a distinct and different pyrogen. The pyrogen obtained from neutrophils had a mol wt of 15,000 following Sephadex G-75 gel filtration, an isoelectric point of 6.9, and could be precipitated and recovered from 50% ethanol at –10°C. In contrast, the pyrogen derived from monocyte preparations had a mol wt of 38,000, an isoelectric point of 5.1, and was destroyed in cold ethanol. Both molecules were unaffected by viral neuraminidase but biologically destroyed at 80°C for 20 min and with trypsin at pH 8.0. The febrile peak produced by partially purified neutrophil pyrogen occurred at 40 min while that from monocytes was at 60 min. In addition, monocyte pyrogen produced more sustained fevers for the same peak elevation as neutrophil pyrogen. These studies demonstrate for the first time two chemically and biologically distinctive pyrogens derived from circulating human white blood cells and have important implications for our understanding of the pathogenesis of fever in man.

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