scholarly journals Inhibiting Effects of Human Plasma and Serum on Neutrophil Random Migration and Chemotaxis

Blood ◽  
1974 ◽  
Vol 44 (6) ◽  
pp. 843-848 ◽  
Author(s):  
H. U. Keller ◽  
M. W. Hess ◽  
H. Cottier
Keyword(s):  
Neutrophil Granulocytes ◽  
Neutrophil Chemotaxis ◽  
Chemotactic Migration ◽  
Experimental Conditions ◽  
Random Migration ◽  
The Past ◽  
Nitroblue Tetrazolium Reduction ◽  
Normal Human ◽  
Tetrazolium Reduction ◽  
Increased Susceptibility

Abstract Various authors have associated increased susceptibility to infectious diseases in certain patients to inhibitors of neutrophil chemotaxis demonstrable in serum or diluted plasma of these patients. The present experiments showed, however, that serum and diluted plasma but not undiluted plasma from normal human donors consistently inhibited chemotactic migration of autologous human neutrophil granulocytes. Therefore, the presence of such inhibitors in the circulating blood can only be assessed by the evaluation of undiluted plasma. The findings suggest that the experimental conditions which have been used in the past to demonstrate such inhibitors in the circulating blood of patients with increased susceptibility to infections are inadequate, and results need reexamination. The inhibitors affect random locomotion and chemotaxis of neutrophil granulocytes but not phagocytosis or the metabolic burst resulting in nitroblue—tetrazolium reduction. On the other hand, phagocytosis of Staphylococcus albus rendered neutrophils chemotactically unresponsive. The significance of so-called cellular defects of neutrophil chemotaxis in such patients is also considered.

scholarly journals Eosinophils do respond to fMLP

Blood ◽  
1987 ◽  
Vol 70 (2) ◽  
pp. 379-383
Author(s):  
M Yazdanbakhsh ◽  
CM Eckmann ◽  
L Koenderman ◽  
AJ Verhoeven ◽  
D Roos
Keyword(s):  
Oxygen Consumption ◽  
Maximal Response ◽  
Free Calcium ◽  
O2 Consumption ◽  
Blood Samples ◽  
Nitroblue Tetrazolium Reduction ◽  
Percoll Gradients ◽  
Normal Human ◽  
Tetrazolium Reduction ◽  
Normal Human Blood

Eosinophils were isolated from normal human blood by separation over Percoll gradients, which resulted in eosinophil suspensions of a purity higher than 95% and recoveries of about 65%. Normal human eosinophils were found to respond to formyl-methionyl-leucyl-phenylalanine (fMLP) at concentrations greater than 10(-7) mol/L with an increase in the concentration of intracellular free calcium, oxygen consumption, nitroblue tetrazolium reduction, and chemiluminescence. The maximal response of eosinophils to fMLP was lower than that of neutrophils isolated from the same blood samples and required at least ten times as much fMLP as was needed for neutrophils. Low fMLP concentrations (approximately 10(-8) mol/L), which in themselves did not stimulate O2 consumption by either eosinophils or neutrophils, primed these cells to respond to a suboptimal concentration of another stimulus. Purification of eosinophils after treatment of whole blood with fMLP showed that these eosinophils had lost their ability to respond to fMLP. We conclude that normal eosinophils do respond to fMLP and that therefore fMLP should not be used to isolate eosinophils.

scholarly journals The biochemical basis of nitroblue tetrazolium reduction in normal human and chronic granulomatous disease polymorphonuclear leukocytes

Blood ◽  
1976 ◽  
Vol 48 (2) ◽  
pp. 309-313 ◽  
Author(s):  
RL Baehner ◽  
LA Boxer ◽  
J Davis
Keyword(s):  
Chronic Granulomatous Disease ◽  
Polymorphonuclear Leukocytes ◽  
Nitroblue Tetrazolium ◽  
Granulomatous Disease ◽  
Biochemical Basis ◽  
Nitroblue Tetrazolium Reduction ◽  
Normal Human ◽  
Nbt Reduction ◽  
Human Polymorphonuclear Leukocytes ◽  
Tetrazolium Reduction

Normal human polymorphonuclear leukocytes (PMN) placed in anaerobic chambers reaching pO2's of less than 5 mm Hg fail to generate O2-, iodinate ingested particles, and stimulate glucose-1–14C oxidation through the hexose monophosphate shunt. The observation that anaerobic cells are incapable of generating O2- or reducing nitroblue tetrazolium (NBT) to formazan supports the idea that NBT reduction in phagocytizing PMN is due exclusively to oxygen-dependent O2- generating oxidase which is deficient in chronic granulomatous disease leukocytes, despite their hyperphagocytic capacity.

scholarly journals Functional and metabolic studies of polymorphonuclear leukocytes in the congenital Pelger-Huet anomaly

Blood ◽  
1980 ◽  
Vol 55 (3) ◽  
pp. 466-469 ◽  
Author(s):  
CA Johnson ◽  
DA Bass ◽  
AA Trillo ◽  
MS Snyder ◽  
LR DeChatelet
Keyword(s):  
Normal Control ◽  
Bactericidal Activity ◽  
Enzyme Activities ◽  
Polymorphonuclear Leukocytes ◽  
Hexose Monophosphate ◽  
Morphological Abnormality ◽  
Random Migration ◽  
Metabolic Defects ◽  
Nitroblue Tetrazolium Reduction ◽  
Tetrazolium Reduction

Abstract Polymorphonuclear leukocytes (PMNL) from two individuals with congenital Pelger-Huet anomaly (PHA) were examined to determine whether functional or metabolic defects accompanied the known morphological abnormality. No abnormalities of the PHA cells, as compared to normal control cells, were found when tested for quantitative leukocyte enzyme activities, nitroblue tetrazolium reduction, hexose monophosphate shunt activity, superoxide production, generation of chemiluminescence, or iodination. The PHA cells, as compared to normal PMNL, demonstrated normal chemotaxis and random migration, as well as bactericidal activity.

scholarly journals Variant of X-Linked Chronic Granulomatous Disease Revealed by a SevereBurkholderia cepaciaInvasive Infection in an Infant

2013 ◽  
Vol 2013 ◽  
pp. 1-5
Author(s):  
Saul Oswaldo Lugo Reyes ◽  
Nizar Mahlaoui ◽  
Carolina Prando ◽  
Lizbeth Blancas Galicia ◽  
Marjorie Hubeau ◽  
...  
Keyword(s):  
Chronic Granulomatous Disease ◽  
Burkholderia Cepacia ◽  
Recurrent Infection ◽  
Granulomatous Disease ◽  
Protein Subunits ◽  
Nitroblue Tetrazolium Reduction ◽  
Bacteria And Fungi ◽  
High Index Of Suspicion ◽  
Tetrazolium Reduction ◽  
Increased Susceptibility

Chronic granulomatous disease (CGD) is a primary immunodeficiency characterized by increased susceptibility to bacteria and fungi since early in life, caused by mutations in any of the five genes coding for protein subunits in NADPH oxidase. X-linked variant CGD can be missed during routine evaluation or present later in life due to hypomorphic mutations and a residual superoxide production. The case of a 10-month-old boy who died of pneumonia is reported. The isolation ofBurkholderia cepaciafrom his lung, together with a marginally low nitroblue tetrazolium reduction assay (NBT), made us suspect and pursue the molecular diagnosis of CGD. A postmortem genetic analysis finally demonstrated CGD caused by a hypomorphic missense mutation with normal gp91phoxexpression. In a patient being investigated for unusually severe or recurrent infection, a high index of suspicion of immunodeficiency must be maintained.

scholarly journals Functional and metabolic studies of polymorphonuclear leukocytes in the congenital Pelger-Huet anomaly

Blood ◽  
1980 ◽  
Vol 55 (3) ◽  
pp. 466-469
Author(s):  
CA Johnson ◽  
DA Bass ◽  
AA Trillo ◽  
MS Snyder ◽  
LR DeChatelet
Keyword(s):  
Normal Control ◽  
Bactericidal Activity ◽  
Enzyme Activities ◽  
Polymorphonuclear Leukocytes ◽  
Hexose Monophosphate ◽  
Morphological Abnormality ◽  
Random Migration ◽  
Metabolic Defects ◽  
Nitroblue Tetrazolium Reduction ◽  
Tetrazolium Reduction

Polymorphonuclear leukocytes (PMNL) from two individuals with congenital Pelger-Huet anomaly (PHA) were examined to determine whether functional or metabolic defects accompanied the known morphological abnormality. No abnormalities of the PHA cells, as compared to normal control cells, were found when tested for quantitative leukocyte enzyme activities, nitroblue tetrazolium reduction, hexose monophosphate shunt activity, superoxide production, generation of chemiluminescence, or iodination. The PHA cells, as compared to normal PMNL, demonstrated normal chemotaxis and random migration, as well as bactericidal activity.

scholarly journals The biochemical basis of nitroblue tetrazolium reduction in normal human and chronic granulomatous disease polymorphonuclear leukocytes

Blood ◽  
1976 ◽  
Vol 48 (2) ◽  
pp. 309-313 ◽  
Author(s):  
RL Baehner ◽  
LA Boxer ◽  
J Davis
Keyword(s):  
Chronic Granulomatous Disease ◽  
Polymorphonuclear Leukocytes ◽  
Nitroblue Tetrazolium ◽  
Granulomatous Disease ◽  
Biochemical Basis ◽  
Nitroblue Tetrazolium Reduction ◽  
Normal Human ◽  
Nbt Reduction ◽  
Human Polymorphonuclear Leukocytes ◽  
Tetrazolium Reduction

Abstract Normal human polymorphonuclear leukocytes (PMN) placed in anaerobic chambers reaching pO2's of less than 5 mm Hg fail to generate O2-, iodinate ingested particles, and stimulate glucose-1–14C oxidation through the hexose monophosphate shunt. The observation that anaerobic cells are incapable of generating O2- or reducing nitroblue tetrazolium (NBT) to formazan supports the idea that NBT reduction in phagocytizing PMN is due exclusively to oxygen-dependent O2- generating oxidase which is deficient in chronic granulomatous disease leukocytes, despite their hyperphagocytic capacity.

Eosinophils do respond to fMLP

Blood ◽  
1987 ◽  
Vol 70 (2) ◽  
pp. 379-383 ◽  
Author(s):  
M Yazdanbakhsh ◽  
CM Eckmann ◽  
L Koenderman ◽  
AJ Verhoeven ◽  
D Roos
Keyword(s):  
Oxygen Consumption ◽  
Maximal Response ◽  
Free Calcium ◽  
O2 Consumption ◽  
Blood Samples ◽  
Nitroblue Tetrazolium Reduction ◽  
Percoll Gradients ◽  
Normal Human ◽  
Tetrazolium Reduction ◽  
Normal Human Blood

Abstract Eosinophils were isolated from normal human blood by separation over Percoll gradients, which resulted in eosinophil suspensions of a purity higher than 95% and recoveries of about 65%. Normal human eosinophils were found to respond to formyl-methionyl-leucyl-phenylalanine (fMLP) at concentrations greater than 10(-7) mol/L with an increase in the concentration of intracellular free calcium, oxygen consumption, nitroblue tetrazolium reduction, and chemiluminescence. The maximal response of eosinophils to fMLP was lower than that of neutrophils isolated from the same blood samples and required at least ten times as much fMLP as was needed for neutrophils. Low fMLP concentrations (approximately 10(-8) mol/L), which in themselves did not stimulate O2 consumption by either eosinophils or neutrophils, primed these cells to respond to a suboptimal concentration of another stimulus. Purification of eosinophils after treatment of whole blood with fMLP showed that these eosinophils had lost their ability to respond to fMLP. We conclude that normal eosinophils do respond to fMLP and that therefore fMLP should not be used to isolate eosinophils.

scholarly journals SpheroidPicker for automated 3D cell culture manipulation using deep learning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Istvan Grexa ◽  
Akos Diosdi ◽  
Maria Harmati ◽  
Andras Kriston ◽  
Nikita Moshkov ◽  
...  
Keyword(s):  
Cell Culture ◽  
Precision Medicine ◽  
High Throughput Screening ◽  
Standard Sample ◽  
Ex Vivo ◽  
Low Cost ◽  
3D Cell Culture ◽  
Syringe Pump ◽  
Experimental Conditions ◽  
The Past

AbstractRecent statistics report that more than 3.7 million new cases of cancer occur in Europe yearly, and the disease accounts for approximately 20% of all deaths. High-throughput screening of cancer cell cultures has dominated the search for novel, effective anticancer therapies in the past decades. Recently, functional assays with patient-derived ex vivo 3D cell culture have gained importance for drug discovery and precision medicine. We recently evaluated the major advancements and needs for the 3D cell culture screening, and concluded that strictly standardized and robust sample preparation is the most desired development. Here we propose an artificial intelligence-guided low-cost 3D cell culture delivery system. It consists of a light microscope, a micromanipulator, a syringe pump, and a controller computer. The system performs morphology-based feature analysis on spheroids and can select uniform sized or shaped spheroids to transfer them between various sample holders. It can select the samples from standard sample holders, including Petri dishes and microwell plates, and then transfer them to a variety of holders up to 384 well plates. The device performs reliable semi- and fully automated spheroid transfer. This results in highly controlled experimental conditions and eliminates non-trivial side effects of sample variability that is a key aspect towards next-generation precision medicine.

Neutrophil chemotaxis, random migration, and adherence in patients with hyperthyroidism

1989 ◽  
Vol 121 (6) ◽  
pp. 817-820 ◽  
Author(s):  
B. Wolach ◽  
B. Lebanon ◽  
A. Jedeikin ◽  
M. S. Shapiro ◽  
L. Shenkman
Keyword(s):  
Thyroid Function ◽  
Chemotactic Activity ◽  
Neutrophil Chemotaxis ◽  
Phagocytic Cells ◽  
Random Migration ◽  
Significant Difference ◽  
Neutrophil Adherence ◽  
Hyperthyroid Patients ◽  
Normal Controls ◽  
Neutrophil Chemotactic Activity

Abstract. We have examined neutrophil adherence, chemotactic activity, and random migration in 35 hyperthyroid patients with Graves' disease and 106 normal volunteers. No statistically significant differences were found between granulocyte adherence of 17 hyperthyroid subjects (67 ± 15.6%) and 81 healthy volunteers (63.1 ± 17%). In 3 thyrotoxic patients, impaired neutrophil adherence was found, which resolved when thyroid function returned to normal. The neutrophil chemotactic activity of 32 normal controls was 107.5 ± 21.4 cells, and the random migration 36 ± 15.5 cells. No statistically significant difference was demonstrated in 13 hyperthyroid patients who had a neutrophil chemotactic activity of 102 ± 14.6 cells and a random migration of 31.2 ± 13.2 cells. Defective chemotactic activity and random migration was found in 2 patients. Neutrophil functions returned to normal in one of the two subjects who were re-evaluated when thyroid function recovered. In summary, 14% of hyperthyroid patients had impaired leukocyte functions. However, severe pyogenic infections are quite rare in hyperthyroid patients, indicating that the observed alterations in function of phagocytic cells are not clinically important.

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