The Relationship between COVID-19 Severity in Children and Immunoregulatory Gene Polymorphism.

Coronavirus disease (COVID-19) and its outcomes remain one of the most challenging problems today. COVID-19 in children could be asymptomatic, but can result in a fatal outcome; therefore, predictions of the disease severity are important. The goal was to investigate the human genetic factors that could be associated with COVID-19 severity in children. Single-nucleotide polymorphisms of the following genes were studied: ACE2 (rs2074192), IFNAR2 (rs2236757), TYK2 (rs2304256), OAS1 (rs10774671), OAS3 (rs10735079), CD40 (rs4813003), FCGR2A (rs1801274) and CASP3 (rs113420705). In the case-control study were 30 children with mild or moderate course of the disease; 30 with severe COVID-19 symptoms and multisystem inflammatory syndrome in children (MIS-C) and 15 who were healthy, and who did not have SARS-CoV-2 (PCR negative, Ig G negative). The study revealed that ACE2 rs2074192 (allele T), IFNAR2 rs2236757 (allele A), OAS1 rs10774671 (allele A), CD40 rs4813003 (allele C), CASP3 rs113420705 (allele C) and male sex contribute to severe COVID-19 course and MIS-C in 85.6% of cases. The World Health Organization reported that new SARS-CoV-2 variants may cause previously unseen symptoms in children. Although the study has limitations due to cohort size, the findings can help provide a better understanding of SARS-CoV-2 infection and proactive pediatric patient management.

Blood ACE2 Protein Level Correlates with COVID-19 Severity.

ACE2's impact on the severity of COVID-19 is widely discussed but still controversial. To estimate its role in aspects of the main risk factors and comorbidities, we involved post-COVID-19 patients in Ternopil region (Ukraine). The recruitment period was from July 2020 to December 2021. Medical records, treatment modalities, and outcomes were recorded and analyzed. The serum human ACE2 protein was measured with Cusabio ELISA kits (Houston, TX, USA). Statistical analysis was performed with SPSS21.0 software (SPSS Inc., Chicago, IL, USA). The level of the ACE2 serum protein was significantly higher (p < 0.001) in patients with mild symptoms compared to a more severe course of the disease, and inversely had changed from 1 to 90 days after recovery. In patients with mild COVID-19, ACE2 levels significantly decreased over time, while among critical patients, it increased by 34.1 percent. Such results could be explained by ACE2 shedding from tissues into circulation. Loss of the membrane-bound form of the enzyme decreases the virus' entry into cells. Our studies did not identify a sex-related ACE2 serum level correlation. The most common comorbidities were hypertension, cardiovascular diseases, respiratory diseases, and diabetes mellitus. All abovementioned comorbidities except respiratory diseases contribute to the severity of the disease and correlate with ACE2 blood serum levels.

Vessel-on-a-Chip: A Powerful Tool for Investigating Endothelial COVID-19 Fingerprints.

Coronavirus disease (COVID-19) causes various vascular and blood-related reactions, including exacerbated responses. The role of endothelial cells in this acute response is remarkable and may remain important beyond the acute phase. As we move into a post-COVID-19 era (where most people have been or will be infected by the SARS-CoV-2 virus), it is crucial to define the vascular consequences of COVID-19, including the long-term effects on the cardiovascular system. Research is needed to determine whether chronic endothelial dysfunction following COVID-19 could lead to an increased risk of cardiovascular and thrombotic events. Endothelial dysfunction could also serve as a diagnostic and therapeutic target for post-COVID-19. This review covers these topics and examines the potential of emerging vessel-on-a-chip technology to address these needs. Vessel-on-a-chip would allow for the study of COVID-19 pathophysiology in endothelial cells, including the analysis of SARS-CoV-2 interactions with endothelial function, leukocyte recruitment, and platelet activation. "Personalization" could be implemented in the models through induced pluripotent stem cells, patient-specific characteristics, or genetic modified cells. Adaptation for massive testing under standardized protocols is now possible, so the chips could be incorporated for the personalized follow-up of the disease or its sequalae (long COVID) and for the research of new drugs against COVID-19.

Influence of chronic hyperhomocysteinemia on the features of bone metabolism in the case of lipopolysaccharide-induced periodontitis.

BACKGROUND: Periodontal disease is the second most common oral health problem after dental caries. This increasing prevalence makes it not only a health problem, but also a social issue. The pathogenesis of periodontal disease is associated with a number of adverse exogenous and endogenous factors, including hyperhomocysteinemia (HHcy). OBJECTIVES: This study aimed to determine the features of bone metabolism in rats with lipopolysaccharide (LPS)-induced periodontitis combined with chronic thiolactone HHcy. MATERIAL AND METHODS: Forty-eight white, non-linear, mature rats were divided into 4 groups: control (n = 12); LPS­induced periodontitis (n = 12); chronic thiolactone HHcy (n = 12); and periodontitis combined with HHcy (n = 12). The rats were sacrificed the day after the last LPS injection or the day after the last homocysteine (Hcy) thiolactone administration. Bone metabolism was determined based on the activity of alkaline phosphatase (ALP) and acid phosphatase (AP) in blood serum and periodontal homogenate. RESULTS: A decrease in ALP activity (by 40.1%; р = 0.001) and the mineralization index (MI) (3.5 times; р < 0.001) with an increase in AP activity (2.0 times; р < 0.001) was observed in the periodontal homogenate of rats with LPS­induced periodontitis. In the case of LPS­induced periodontitis combined with chronic thiolactone HHcy, more pronounced changes in the activity of phosphatases and in MI were established as compared to rats with LPS­induced periodontitis only. CONCLUSIONS: Chronic thiolactone HHcy enhances disturbances in bone metabolism in LPS­induced periodontitis. The osteotoxic effect of HHcy is associated with the activation of osteoclastogenesis and enhanced bone resorption. However, further research is required on the subject.

Immunoregulatory Intestinal Microbiota and COVID-19 in Patients with Type Two Diabetes: A Double-Edged Sword.

Coronavirus disease 2019, or COVID-19, is a major challenge facing scientists worldwide. Alongside the lungs, the system of organs comprising the GI tract is commonly targeted by COVID-19. The dysbiotic modulations in the intestine influence the disease severity, potentially due to the ability of the intestinal microbiota to modulate T lymphocyte functions, i.e., to suppress or activate T cell subpopulations. The interplay between the lungs and intestinal microbiota is named the gut-lung axis. One of the most usual comorbidities in COVID-19 patients is type 2 diabetes, which induces changes in intestinal microbiota, resulting in a pro-inflammatory immune response, and consequently, a more severe course of COVID-19. However, changes in the microbiota in this comorbid pathology remain unclear. Metformin is used as a medication to treat type 2 diabetes. The use of the type 2 diabetes drug metformin is a promising treatment for this comorbidity because, in addition to its hypoglycemic action, it can increase amount of intestinal bacteria that induce regulatory T cell response. This dual activity of metformin can reduce lung damage and improve the course of the COVID-19 disease.

"All that glitters is not gold": sialomegaly, xerostomia, and dry eye is not always Sjögren's syndrome.

Although dry mouth, dry eye, and swollen salivary glands are the hallmark manifestations of Sjögren's syndrome (pSS), a wide spectrum of other conditions should be considered for differential diagnosis. The diagnostic challenge is mainly encountered in patients presenting with dry eyes and/or dry mouth, who do not meet the full established classification criteria for pSS. Presented case-based review highlights the relationship between dry eye, parotid swelling, and psychiatric disorders. The obsessive-compulsive disorder may be separately be a cause of dryness symptoms even were not using any psychiatric drugs. The presented review widely discussed this problem and the aim is to shed new light on the interpretation of the dryness symptom and the necessity of individual patient assessment, excluding causes other than pSS before making a final diagnosis and making a decision on the treatment method.

Chitosan self-assembled polymeric nanoparticles for percutaneous delivery of betamethasone in contact dermatitis.

OBJECTIVE: The study was performed with an aim to investigate the efficiency of two treatment options in experimental nickel-induced contact dermatitis (CT), with either betamethasone or chitosan cross-linked nano-encapsulated betamethasone lanoline solutions (nano-betamethasone). METHODS: Male Wistar rats were used. The differences were compared based on lesion visual appearance, skinfold thickness, white blood cell count (WBC), erythrocyte sedimentation rate (ESR), blood serum prooxidant-antioxidant balance (thiobarbituric acid reactive substances, TBARS; supersoxide dismutase, SOD; catalase, KAT), blood cytokine profile (TNF-α, IL-1ß, IL-10, and IL-4), and histological examination of affected skin. RESULTS: All animals treated with nickel sulfate developed CT and systemic inflammatory response on day 12, which only slightly lessened, if left untreated, on day 20. The therapeutic effectiveness of nano-betamethasone was significantly far superior (p < 0.01) compared to betamethasone. Specifically, the visual appearance of lesion severity of betamethasone vs. nano-betamethasone ± SD was 1.82 ± 0.18 vs. 1.17 ± 0.24 points, skinfold thickness-2.68 ± 0.12 vs. 2.12 ± 0.10 mm, ESR-6.38 ± 0.27 vs. 5.12 ± 0.20 mm/h, WBC-8.47 ± 0.28 vs. 7.17 ± 0.24 109/L, TBARS-1.09 ± 0.04 vs. 0.94 ± 0.02 µmol/L, SOD-3.38 ± 0.26 vs. 4.12 ± 0.18 r.u./L, KAT-11.54 ± 0.14 vs. 10.02 ± 0.19 mkatal/L, respectively. The nano-betamethasone formulation was also more effective (p < 0.01) in increasing anti-inflammatory cytokines level, IL-10 (8.96 ± 0.32 vs. 7.54 ± 0.52 pg/mL) and IL-4 (13.16 ± 0.45 vs. 11.43 ± 0.58 pg/mL); and decreasing in pro-inflammatory TNF-α (20.94 ± 2.30 vs. 26.98 ± 1.16 pg/mL) and IL-1ß (19.35 ± 1.28 vs. 24.77 ± 1.75 pg/mL), respectively. These findings were also supported with histological examination. CONCLUSIONS: Nano-betamethasone may be considered as a more successful transcutaneous therapy for managing contact dermatitis compared to ointments consisting of betamethasone in traditional form.

Metformin Attenuates Postinfarction Myocardial Fibrosis and Inflammation in Mice.

Diabetes is a major risk factor for the development of cardiovascular disease with a higher incidence of myocardial infarction. This study explores the role of metformin, a first-line antihyperglycemic agent, in postinfarction fibrotic and inflammatory remodeling in mice. Three-month-old C57BI/6J mice were submitted to 30 min cardiac ischemia followed by reperfusion for 14 days. Intraperitoneal treatment with metformin (5 mg/kg) was initiated 15 min after the onset of reperfusion and maintained for 14 days. Real-time PCR was used to determine the levels of COL3A1, αSMA, CD68, TNF-α and IL-6. Increased collagen deposition and infiltration of macrophages in heart tissues are associated with upregulation of the inflammation-associated genes in mice after 14 days of reperfusion. Metformin treatment markedly reduced postinfarction fibrotic remodeling and CD68-positive cell population in mice. Moreover, metformin resulted in reduced expression of COL3A1, αSMA and CD68 after 14 days of reperfusion. Taken together, these results open new perspectives for the use of metformin as a drug that counteracts adverse myocardial fibroticand inflammatory remodeling after MI.

Does thyroid dysfunction influence inflammatory mediators in experimental periodontitis?

Objective. The aim of the present study was to investigate the presence of inflammatory mediators in rats with only periodontitis and periodontitis in a setting of hyper- and hypo-thyroidism and to analyze the correlative linkages between inflammatory mediators and thyroid hormones. Methods. White male 12-14 weeks old inbred rats (n=48) weighing 180-200 g were employed in the experiment. They were randomly divided into the following groups: Group I - control group, Group II - group with a model of periodontitis, Group III - group with a periodontitis in a setting of hyperthyroidism, and Group IV - group with periodontitis in a setting of hypothyroidism. The presence of tumor-necrosis factor-α (TNF-α) and interleukins IL-1ß and IL-10 in the periodontal homogenate supernatant was studied by a solid-phase enzyme-linked immunosorbent assay. Results. It was shown that experimental lipopolysaccharide (LPS)-induced periodontitis is accompanied by hyperproduction of pro-inflammatory cytokines (TNF-α, IL-1ß) and reduction of anti-inflammatory cytokines (IL-10), whereas TNF-α underwent to maximum changes. Thyroid dysfunction exacerbates cytokine imbalance and severity of inflammation in experimental LPS-induced periodontitis, especially pronounced at hyperthyroidism, as evidenced by the predominance of TNF-α and IL-1ß levels in the periodontal homogenate supernatant by 38.5% (р<0.01) and 75.6% (p<0.001), respectively, hyperthyroid over the euthyroid, and by 20.1% (p<0.05) and 24.1% (p<0.05), respectively, over the hypothyroid rats. Conclusions. Thyroid dysfunction, especially hyperthyroidism, may play an important role in the pro-inflammatory response in periodontitis. Hyperproduction of inflammatory mediators in thyroid dysfunction can induce a noticeable damage in the whole apparatus of the periodontium, thereby causing progression of periodontitis.

A Comparative Study of Connective Tissue Metabolism Indices in Experimental Comorbidity-Free Periodontitis and Periodontitis Combined with Thyroid Dysfunction.

Periodontal disease is a chronic bacterial infection characterized by persistent inflammation, connective tissue breakdown, and alveolar bone destruction. The current study aimed to compare the connective tissue metabolism indices in rats with comorbidity-free periodontitis and in animals with periodontitis in a setting of hyper-and hypothyroidism. 12-14-week-old inbred white male rats (n=48) were included in the experiment. They were randomly divided into the following groups: control, animals with a model of periodontitis, animals with periodontitis in a setting of hyperthyroidism, animals with periodontitis in a setting of hypothyroidism. Serum levels of free thyroxine, free triiodothyronine, and thyroid-stimulating hormone were assayed using ELISA kits manufactured by Vector Best (Russia) to confirm the hyper- and hypothyroid status. Collagenolytic activity, the content of glycosaminoglycans, free hydroxyproline, and fucose, unbound with proteins in blood serum were assayed using the spectrophotometric method. We have found the increasing of collagenolytic activity by 46.1% (р<0.001), the content of free hydroxyproline by 74.1% (р<0.001), the content of glycosaminoglycans by 1.8 times (р<0.001), the content of fucose, unbound with proteins by 2.8 times (р<0.001) in rats with periodontitis vs. the control group. The development of periodontitis in a setting of thyroid dysfunction leads to an even more significant increase in the destruction of connective tissue, which is confirmed by a significant increase in the content of studied indices vs. euthyroid animals, both in hyperthyroidism and hypothyroidism.

Nerve impulse transmission pathway-focused genes expression analysis in patients with primary hypothyroidism and autoimmune thyroiditis.

OBJECTIVE: Thyroid hormones have important actions in the adult brain. They regulate genes expression in myelination, differentiation of neuronal and glial cells, and neuronal viability and function. METHODS: We used the pathway-specific real-time PCR array (Neurotrophins and Receptors RT2 Profiler PCR Array, QIAGEN, Germany) to identify and verify nerve impulse transmission pathway-focused genes expression in peripheral white blood cells of patients with postoperative hypothyroidism, hypothyroidism as a result of autoimmune thyroiditis (AIT) and AIT with elevated serum an anti-thyroglobulin (anti-Tg) and anti-thyroid peroxidase (anti-TPO) antibodies. RESULTS: It was shown that patients with postoperative hypothyroidism and hypothyroidism resulting from AIT had significantly lower expression of BDNF and CBLN1. In patients with AIT with elevated serum anti-Tg and anti-TPO antibodies, the expression of GDNF was significantly down-regulated and the expression of PNOC was up-regulated. The expression levels of MEF2C and NTSR1 were decreased in the group of patients with postoperative hypothyroidism and AIT, correspondingly. CONCLUSIONS: The results of this study demonstrate that AIT and hypothyroidism can affect the expression of mRNA nerve impulse transmission genes in gene specific manner and that these changes in gene expressions can be playing a role in the development of neurological complications associated with thyroid pathology. Detection of the transcriptional activity of nerve impulse transmission genes in peripheral white blood cells can be used as an important minimally invasive prognostic marker of the risk for developing neurological complications comorbid with thyroid pathology.

A comparative study of oral microbiocenosis structure in experimental comorbidity-free periodontitis and in periodontitis combined with thyroid dysfunction.

Thyroid hormones regulate numerous metabolic processes. Therefore, any alteration in their synthesis or function has important health implications. However, limited data are available regarding the relationship between thyroid hormone imbalance and periodontal health. AIM: The aim of the study was to perform a comparative analysis of qualitative and quantitative structure of oral microbiocenosis in rats with comorbidity-free periodontitis and in animals with periodontitis in a setting of hyper- and hypothyroidism. MATERIALS AND METHODS: Inbred white male rats (n=48) were randomly divided into the following groups: I - control animals, II - animals with a model of periodontitis, III - rats with periodontitis in a setting of hyperthyroidism, IV - rats with periodontitis in a setting of hypothyroidism. Samples for microbiological investigations were taken from dental surfaces (on the border between hard tissue and gums in the interdental spaces). The isolated pure cultures were identified by their morphological, tinctorial, cultural and biochemical properties. RESULTS: The oral dysbiosis occurring in a setting of periodontitis in rats is chiefly characterized by increased quantity of coccal forms and by increased candidal inoculation; these organisms cumulatively inhibit the growth of normal microbial flora, such as Lactobacilli, bacteroids and Bifidobacteria. The periodontitis in a setting of thyroid dysfunction increases both the species variety and the quantitative counts of oral microbial flora, with predominance of such microbial organisms as Staph. aureus, E. coli, E. faecalis, Candida albicans and P. aeruginosa. Comparative assessment of intensity of oral microbial colonization in hyper- and hypothyroid animals with periodontitis has demonstrated significant changes only for the strains of S. aureus, yeast-like fungi and Candida albicans, which were predominant in hyperthyroid rats. CONCLUSIONS: Both hyperthyroidism and hypothyroidism exacerbate changes in the qualitative and quantitative structure of oral microbiocenosis in case of experimental periodontitis.

Line Immunoblot Assay for Tick-Borne Relapsing Fever and Findings in Patient Sera from Australia, Ukraine and the USA.

Tick-borne relapsing fever (TBRF) is caused by spirochete bacteria of the genus Borrelia termed relapsing fever Borreliae (RFB). TBRF shares symptoms with Lyme disease (LD) caused by related Lyme disease Borreliae (LDB). TBRF and LD are transmitted by ticks and occur in overlapping localities worldwide. Serological detection of antibodies used for laboratory confirmation of LD is not established for TBRF. A line immunoblot assay using recombinant proteins from different RFB species, termed TBRF IB, was developed and its diagnostic utility investigated. The TBRF IBs were able to differentiate between antibodies to RFB and LDB and had estimated sensitivity, specificity, and positive and negative predictive values of 70.5%, 99.5%, 97.3%, and 93.4%, respectively, based on results with reference sera from patients known to be positive and negative for TBRF. The use of TBRF IBs and analogous immunoblots for LD to test sera of patients from Australia, Ukraine, and the USA with LD symptoms revealed infection with TBRF alone, LD alone, and both TBRF and LD. Diagnosis by clinical criteria alone can, therefore, underestimate the incidence of TBRF. TBRF IBs will be useful for laboratory confirmation of TBRF and understanding its epidemiology worldwide.

Metformin Protects the Heart Against Hypertrophic and Apoptotic Remodeling After Myocardial Infarction.

Cardiovascular complications are the most prevalent cause of morbidity and mortality in diabetic patients. Metformin is currently the first-line blood glucose-lowering agent with potential relevance to cardiovascular diseases. However, the underpinning mechanisms of action remain elusive. Here, we report that metformin represses cardiac apoptosis at least in part through inhibition of Forkhead box O1 (FoxO1) pathway. In a mouse model of ischemia-reperfusion (I/R), treatment with metformin attenuated cardiac and hypertrophic remodeling after 14 days of post-reperfusion. Additionally, cardiac expression of brain-like natriuretic peptide (BNP) was significantly reduced in metformin-treated mice after 14 days of cardiac I/R. In cultured H9C2 cells, metformin counteracted hypertrophic and apoptotic responses to metabolic or hypoxic stress. FoxO1 silencing by siRNA abolished anti-apoptotic effect of metformin under hypoxic stress in H9C2 cells. Taken together, these results suggest that metformin protects the heart against hypertrophic and apoptotic remodeling after myocardial infarction.

Leptin-induced endothelial dysfunction in obesity.

Hyperleptinemia accompanying obesity affects endothelial nitric oxide (NO) and is a serious factor for vascular disorders. NO, superoxide (O(2)(-)), and peroxynitrite (ONOO(-)) nanosensors were placed near the surface (5+/-2 microm) of a single human umbilical vein endothelial cell (HUVEC) exposed to leptin or aortic endothelium of obese C57BL/6J mice, and concentrations of calcium ionophore (CaI)-stimulated NO, O(2)(-), ONOO(-) were recorded. Endothelial NO synthase (eNOS) expression and L-arginine concentrations in HUVEC and aortic endothelium were measured. Leptin did not directly stimulate NO, O(2)(-), or ONOO(-) release from HUVEC. However, a 12-h exposure of HUVEC to leptin increased eNOS expression and CaI-stimulated NO (625+/-30 vs. 500+/-24 nmol/l control) and dramatically increased cytotoxic O(2)(-) and ONOO(-) levels. The [NO]-to-[ONOO(-)] ratio ([NO]/[ONOO(-)]) decreased from 2.0+/-0.1 in normal to 1.30+/-0.1 in leptin-induced dysfunctional endothelium. In obese mice, a 2.5-fold increase in leptin concentration coincided with 100% increase in eNOS and about 30% decrease in intracellular L-arginine. The increased eNOS expression and a reduced l-arginine content led to eNOS uncoupling, a reduction in bioavailable NO (250+/-10 vs. 420+/-12 nmol/l control), and an elevated concentration of O(2)(-) (240%) and ONOO(-) (70%). L-Arginine and sepiapterin supplementation reversed eNOS uncoupling and partially restored [NO]/[ONOO(-)] balance in obese mice. In obesity, leptin increases eNOS expression and decreases intracellular l-arginine, resulting in eNOS an uncoupling and depletion of endothelial NO and an increase of cytotoxic ONOO(-). Hyperleptinemia triggers an endothelial NO/ONOO(-) imbalance characteristic of dysfunctional endothelium observed in other vascular disorders, i.e., atherosclerosis and diabetes.

Effect of beta-blockers on endothelial function during biological aging: a nanotechnological approach.

Biological aging is an independent risk factor for many cardiovascular diseases; some are treated with beta-blockers that may protect dysfunctional endothelium during aging by increasing NO, decreasing ONOO, and restoring NO/ONOO balance. A nanotechnological approach was used to simultaneously monitor NO and ONOO produced by a single aortic endothelial cell from Wistar-Kyoto rats of different ages. beta-blockers (metoprolol and atenolol) were administered 2 weeks before the animals were sacrificed. Nanosensors were placed near the endothelium, and calcium ionophore- stimulated NO and ONOO release was measured. Endothelial nitric oxide synthase (eNOS) undergoes uncoupling with aging, manifested by a decrease in NO (from 503 +/- 12 to 163 +/- 5 nmol/L) and a 3-fold increase in ONOO for 16-week-old and 110- week- old rats, respectively. Metoprolol reversed eNOS uncoupling, increased the production rate and concentration of NO, and increased an overall ratio of NO]/ONOO]. This effect was not observed with atenolol, but L-arginine, sepiapterin, and superoxide dismutase were beneficial.

Effect of aspirin on constitutive nitric oxide synthase and the biovailability of NO.

INTRODUCTION: Aspirin decreases the activity of iNOS and the formation of prostanoids. Constitutive nitric oxide synthase (cNOS) is present in endothelial cells, platelets, leukocytes and neurons, yet no data are available on the effect of aspirin on cNOS and the bioavailability of NO produced by this enzyme. MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVECs), rat adrenal gland pheochromocytoma cells (PC-12) and human platelets were incubated with different aspirin concentrations. The kinetics of NO, O2- and ONOO- release were measured simultaneously in single cells or platelet suspensions using tandem electrochemical nanosensors. The NO, O2- and ONOO- release from cells and platelets was stimulated with calcium ionophore and collagen, respectively. cNOS expression was estimated by Western blot analysis. RESULTS: Incubation of HUVECs and PC-12 with 10(-5) mol/l of aspirin increased cNOS expression by 70 +/- 7% and 50 +/- 5, respectively. However, the NO concentration increased only by 33% in HUVECs incubated with the same aspirin concentration. Incubation of HUVECs with aspirin also increased the O2- and ONOO- production. Therefore the bioavailability of NO increased only slightly in endothelium and did not reflect the increase in eNOS. This was in contrast to platelets, where maximal NO bioavailability almost doubled after incubation with aspirin. CONCLUSIONS: Aspirin did not have a significant effect on the NO bioavailability in endothelial cells. However, aspirin highly improved the NO production in platelets. The high NO production in platelets may counteract the effect of thromboxane, inhibit platelet aggregation, and compensate for the reduction of prostacycline concentration by aspirin.