Comprehensive Assessment of Mid-Regional Proadrenomedullin, Procalcitonin, Neuron-Specific Enolase and Protein S100 for Predicting Pediatric Severe Trauma Outcomes.

The development of multiple organ failure and septic complications increases the cumulative risk of mortality in children with severe injury. Clinically available biochemical markers have shown promise in assessing the severity and predicting the development of complications and outcomes in such cases. This study aimed to determine informative criteria for assessing the severity and outcome prediction of severe injury in children based on levels of mid-regional proadrenomedullin (MR-proADM) procalcitonin (PCT), neuron-specific enolase (NSE), and protein S100. Biomarker levels were measured in 52 children with severe injury (ISS ≥ 16) on the 1st, 3rd, 7th, and 14th days after admission to the ICU. The children were divided into groups based on their favorable (n = 44) or unfavorable (n = 8) outcomes according to the Severe Injury Outcome Scale, as well as their favorable (n = 35) or unfavorable (n = 15) outcomes according to the Glasgow Coma Outcome Scale (GOS). The study also evaluated the significance of biomarker levels in predicting septic complications (with SC (n = 16) and without SC (n = 36)) and diagnosing and stratifying multiple organ failure (with MOF (n = 8) and without MOF (n = 44)). A comprehensive assessment of MR-proADM and PCT provided the highest diagnostic and prognostic efficacy for early diagnosis, risk stratification of multiple organ failure, and outcome prediction in severe injury cases involving children. Additionally, the inclusion of the S100 protein in the study allowed for further assessment of brain damage in cases of traumatic brain injury (TBI), contributing to the overall prognostic model.

Quantitative Analysis of Latex Beads Phagocytosis by Human Macrophages Using Imaging Flow Cytometry with Extended Depth of Field (EDF).

The existing methods of quantitative analysis of phagocytosis are characterized by a number of limitations. The usual method of manually counting phagocytosed objects on photographs obtained by confocal microscopy is very labor-intensive and time-consuming. As well, the resolution of conventional flow cytometry does not allow the fluorescence detection of a large number of phagocytosis objects. Thus, there is a need to combine the rapid analysis by flow cytometry and the visualization capability by confocal microscopy. This is possible due to imaging flow cytometry. However, until now, no protocols have allowed one to quantify phagocytosis at its high intensity. The present paper presents the developed and tested algorithm for assessing the level of phagocytic activity using flow cytometry with visualization and IDEAS software.

Cell-Molecular Interactions of Nano- and Microparticles in Dental Implantology.

The role of metallic nano- and microparticles in the development of inflammation has not yet been investigated. Soft tissue biopsy specimens of the bone bed taken during surgical revisions, as well as supernatants obtained from the surface of the orthopedic structures and dental implants (control), were examined. Investigations were performed using X-ray microtomography, X-ray fluorescence analysis, and scanning electron microscopy. Histological studies of the bone bed tissues were performed. Nanoscale and microscale metallic particles were identified as participants in the inflammatory process in tissues. Supernatants containing nanoscale particles were obtained from the surfaces of 20 units of new dental implants. Early and late apoptosis and necrosis of immunocompetent cells after co-culture and induction by lipopolysaccharide and human venous blood serum were studied in an experiment with staging on the THP-1 (human monocytic) cell line using visualizing cytometry. As a result, it was found that nano- and microparticles emitted from the surface of the oxide layer of medical devices impregnated soft tissue biopsy specimens. By using different methods to analyze the cell-molecule interactions of nano- and microparticles both from a clinical perspective and an experimental research perspective, the possibility of forming a chronic immunopathological endogenous inflammatory process with an autoimmune component in the tissues was revealed.

A link between mitotic defects and mitotic catastrophe: detection and cell fate.

Although the phenomenon of mitotic catastrophe was first described more than 80 years ago, only recently has this term been used to explain a mechanism of cell death linked to delayed mitosis. Several mechanisms have been suggested for mitotic catastrophe development and cell fate. Depending on molecular perturbations, mitotic catastrophe can end in three types of cell death, namely apoptosis, necrosis, or autophagy. Moreover, mitotic catastrophe can be associated with different types of cell aging, the development of which negatively affects tumor elimination and, consequently, reduces the therapeutic effect. The effective triggering of mitotic catastrophe in clinical practice requires induction of DNA damage as well as inhibition of the molecular pathways that regulate cell cycle arrest and DNA repair. Here we discuss various methods to detect mitotic catastrophe, the mechanisms of its development, and the attempts to use this phenomenon in cancer treatment.

Lipopolysaccharide From E. coli Increases Glutamate-Induced Disturbances of Calcium Homeostasis, the Functional State of Mitochondria, and the Death of Cultured Cortical Neurons.

Lipopolysaccharide (LPS), a fragment of the bacterial cell wall, specifically interacting with protein complexes on the cell surface, can induce the production of pro-inflammatory and apoptotic signaling molecules, leading to the damage and death of brain cells. Similar effects have been noted in stroke and traumatic brain injury, when the leading factor of death is glutamate (Glu) excitotoxicity too. But being an amphiphilic molecule with a significant hydrophobic moiety and a large hydrophilic region, LPS can also non-specifically bind to the plasma membrane, altering its properties. In the present work, we studied the effect of LPS from Escherichia coli alone and in combination with the hyperstimulation of Glu-receptors on the functional state of mitochondria and Ca2+ homeostasis, oxygen consumption and the cell survival in primary cultures from the rats brain cerebellum and cortex. In both types of cultures, LPS (0.1-10 µg/ml) did not change the intracellular free Ca2+ concentration ([Ca2+]i) in resting neurons but slowed down the median of the decrease in [Ca2+]i on 14% and recovery of the mitochondrial potential (ΔΨm) after Glu removal. LPS did not affect the basal oxygen consumption rate (OCR) of cortical neurons; however, it did decrease the acute OCR during Glu and LPS coapplication. Evaluation of the cell culture survival using vital dyes and the MTT assay showed that LPS (10 µg/ml) and Glu (33 µM) reduced jointly and separately the proportion of live cortical neurons, but there was no synergism or additive action. LPS-effects was dependent on the type of culture, that may be related to both the properties of neurons and the different ratio between neurons and glial cells in cultures. The rapid manifestation of these effects may be the consequence of the direct effect of LPS on the rheological properties of the cell membrane.

Effect of NSAIDs on Pupil Diameter and Expression of Aqueous Humor Cytokines in FLACS Versus Conventional Phacoemulsification.

PURPOSE: To compare the concentrations of interleukin (IL) (IL-1b, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70), interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and TNF-ß in the aqueous humor of patients undergoing femtosecond laser-assisted cataract surgery (FLACS) and corneal mechanical paracentesis treated with two different topical nonsteroidal anti-inflammatory drugs (NSAIDs): bromfenac and indomethacin. METHODS: In this prospective, randomized controlled, single-center study, aqueous humor samples were obtained immediately after performing the femtosecond laser procedure or at the start of conventional phacoemulsification. Preoperatively, the FLACS groups were administered (once daily and four times daily, respectively) either topical bromfenac 0.09% (12 eyes) or indomethacin 0.1% (12 eyes). The corneal paracentesis bromfenac and indomethacin groups received the same regimen of instillation of NSAIDs, respectively. Quantitative analysis of the expressed cytokines in the aqueous humor was performed using FlowCytomix FC 500 Pro 3.0 Software (Bender MedSystems GmbH, Vienna, Austria). RESULTS: The intraoperative pupil diameter was correlated with the expression of IL-6 after the femtosecond laser procedure in the FLACS indomethacin group (r = -0.53; P = .07). A significant difference in mean pupillary size was detected between the FLACS bromfenac and indomethacin groups at the aspiration/irrigation time point (0.53 ± 0.26 mm) and at the end of surgery (0.68 ± 0.37 mm). Progressive pupillary constriction was observed in the indomethacin and bromfenac groups. CONCLUSIONS: A smaller expression of IL-6 to the overall cytokine network value was observed in cases receiving preoperative bromfenac 0.09%, explaining improved maintenance of intraoperative mydriasis. [J Refract Surg. 2018;34(10):646-652.].