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Rationale: An important element of antiviral defense in the pathophysiology of COVID-19 is the innate cell immunity including polymorphonuclear neutrophils prone to netotic transformation. Neutrophils can be not only a marker of acute infection, but, being a source of neutrophil extracellular traps (NET), can play a key role in the development of thrombotic complications leading to acute respiratory insufficiency in COVID-19.Aim: To determine the diagnostic and prognostic value of NET levels in patients with COVID-19.Materials and methods: We monitored NET levels in peripheral blood of 34 patients with COVID-19 (mean age, 67 ± 15.8 years), admitted to MONIKI hospital. The control group consisted of 54 healthy volunteers (mean age, 52 ± 11.5 years). Whole blood samples of 2 μL each were used for the preparation of monolayer smears (Giemsa stain) and calculation of at least 200 cell structures including native intact and transformed neutrophils (MECOS-C2 microscope, Medical computer systems).Results: Patients with COVID-19 had higher NET levels, compared to those in healthy controls: 14.5% (2.9–28.6%) vs. 5.0% (1.8–11.9%, p < 0.0001). The patients who were on non-invasive respiratory support (23.5%) had a NET level of 12% (8.1– 22.3%), whereas those on invasive mechanical ventilation (17.6%) had a 1.5-fold higher NET level of 17.9% (12.3–28.2%) (p < 0.05). In the patients who died (11.8% of the cases), the NET level amounted to 19% (16.5–26%, p < 0.05). Monitoring of blood NET levels was performed in 9 patients from the day of admittance to the day of their transfer to the intensive care unit / discharge / death. It was shown that a decrease of NET levels mirrors an improvement of the patient’s clinical condition and efficacy of his/hers treatment. On the opposite, an increase of NET levels can indicate a deterioration and risk of unfavorable course.Conclusion: We have identified some pathophysiological mechanisms in COVID-19, related to the neutrophil compartment. Patients with coronavirus infection are characterized by high NET levels which is at least 3-fold higher than that in healthy volunteers. This indicates an abnormality in immune host defense and development of an inadequate inflammatory response. An increase of NET in whole blood smears of more than 16% can be a criterion of an unfavorable prognosis of the disease course and the risk of death.

The paper presents a review of current techniques of the interference microscopy of living functioning cells that allow for assessment of the real-time changes of subcellular microstructures with 2D and 3D reconstructions of the images and multifactorial data analysis. It has been shown that new diagnostic information can be obtained within the research methodology of the so-called interactive dialogue with the cell to identify changes in the morphofunctional state of living cells through registration of their realtime response to environmental fluctuations. We describe physical basis of laser interferometry, problems and specifics of interpretation of cell phase pictures. As an illustration to the practical use of the technique in the clinic we present the results of studies on morphofunctional state of peripheral blood platelets in patients with oral malignancies (OM) and various stages of the tumor. Analysis of circulating platelets with a realtime technique of laser interferometry allowed for identification of compensated activation of the platelet compartment of hemostasis in these patients, as well as an association between the severity of cancer and changes of dimensional cell parameters (an increase of the mean diameter and area of circulating platelets by 23.4, 26.8 and 30.3%, and by 30.2, 32.5 and 38.1% in the patients with OM grades II, III and IV, respectively (p < 0,05 for all comparisons)). The necessary condition for a wider implementation of new diagnostic technologies is a critical evaluation of the efficacy of their use in routine practice, identification of their advantages, disadvantages and differences between current and proposed diagnostic platforms.

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