Abstract
The previous slide-glass type system could simultaneously detect reactive and highly reactive oxygen species, i.e., superoxide radicals (O2−·) and hypochlorite ions (OCl−) elicited from leucocytes in sample blood, but had some drawbacks, i.e., signal noise from air-flow stirring, potential biohazard risks, etc. because of open samples placed on a slide glass. We overcame these drawbacks by adopting a fluidic-chip container in a new system, which resulted in higher sensitivity and more stable measurements. Using the new system, we conducted a pilot study on nominally healthy volunteers to find whether or not the monitored activities of leukocytes can distinguish more or less unhealthy conditions from healthy ones. At first, healthy volunteers of both genders and of various ages showed that the fluctuation magnitudes (%) of O2−· and OCl− were nearly similar to each other and to that of the neutrophil count fluctuation. These parameters sometimes exceeded the healthy fluctuation range. By comparing these large fluctuations with the data of an inflammation marker C-reactive protein (CRP), the neutrophil count fluctuation and the timings/symptoms of abnormalities found in questionnaire, we could gain information suggesting the factors causing the large fluctuations. The new system could detect bodily abnormalities earlier than CRP or self-aware symptoms.
Highlights
The previous slide-glass type system could simultaneously detect reactive and highly reactive oxygen species, i.e., superoxide radicals (O2−·) and hypochlorite ions (OCl−) elicited from leucocytes in sample blood, but had some drawbacks, i.e., signal noise from air-flow stirring, potential biohazard risks, etc. because of open samples placed on a slide glass
With the new fluidic-chip based system, we could obtain larger and more robust O 2−· and OCl− signals than those obtained with the previous glass slide-type system (CFL-P2200)
In “Results”, we have shown the following two important outcomes: (1) a new simultaneous CL/FL monitoring system (CFL-H2200/2400) was confirmed to be optimized for stable blood sample measurements with higher sensitivity for a very long period of time (Figs. 1, 2, Fig. S1). (2) We were able to clarify the fluctuating ranges of O2−· and OCl− production in a healthy state and the relationship between the changes in physical condition and their production, which provides insight into the question of whether or not the leukocyte activity can distinguish between more or less unhealthy states and healthy ones
Summary
The previous slide-glass type system could simultaneously detect reactive and highly reactive oxygen species, i.e., superoxide radicals (O2−·) and hypochlorite ions (OCl−) elicited from leucocytes in sample blood, but had some drawbacks, i.e., signal noise from air-flow stirring, potential biohazard risks, etc. because of open samples placed on a slide glass. We have recently developed CFL-P22009 and demonstrated that it can semi-automatically detect ROS and hROS, i.e., O 2−· and O Cl− elicited in a small amount of blood samples by using CL and FL detecting reagents. This newer system was recently applied to studying the correlative relationships between O2−·/OCl− and diseases, such as Alzheimer’s disease10, arteriosclerosis[11], and hypertension[12] with the rodent models. Fine-tuning of the air flows (amounts, timing and directions) requires some practice and training
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