The Covid-crisis and the consumption of alcohol-based disinfectants - data and key measures in a critical period

The Covid-19 syndemic shocked the healthcare systems on the old continent. In Europe, including Bulgaria, hospital institutions went through unprecedented difficulties due to the specific reorganization required to achieve an adequate response related to Covid-19. The aim of the study is to present hospital-level data from University Hospital "Sveta Marina" in Varna, regarding the usage of alcohol-based disinfectants (ABDs), comparing the consumption before and during the Covid-19 period. The key measures implemented by the medical institution during the critical period are also systematized.Material and Methods: A single-center retrospective observational study of data from the hospital information system related to the consumption of ABDs. Documentary method, descriptive statistical analysis, and graphical method were used.Results: The data from the current study shows that the overall hospital usage of ABDs has increased during the Covid period (2020-2021), from 8.82 ml per patient day (PD) in 2018 to 16.29 ml per PD. The quantities of ABDs used in the intensive care units have also increased during the Covid period, reaching 76.5 ml per PD for the Clinic of Anesthesiology and Intensive Care, 96.8 ml per PD for the Pediatric Intensive Care Unit, and values in the Intensive Care and Invasive Ventilation Unit have tripled, reaching 121 ml per PD in 2021.Conclusions: In a national context, this is the first study that provides openly data with the idea of provoking developments towards continuing medical and health personnel education for hand hygiene quality. Successful practices at the institutional hospital level are shared, which increase staff compliance

Ethanol induces IL-6 and TNF-α cytokine and iNOS and COX-2 enzyme gene expression in 3T3-L1 preadipocytes

t is known that alcohol influences on human health and its effects are widely studied. Adipose tissue (AT) may be an important target of ethanol action. Data about the effect of ethanol on cytokine and pro-inflammatory enzymes` gene expression in adipocytes are scarce. The aim of the present study was to establish its effect on transcription of inflammatory markers IL-6, TNF-α, COX-2 and iNOS in cultured 3T3-L1 cells. MTT test was performed in order to determine ethanol effect on cell viability. Expression of selected genes was measured using two step quantitative PCR. Results were analyzed using 2-ΔΔCt method. Exposure of 3T3-L1 cells to increasing ethanol concentrations (0.125%-0.625%) resulted in gradual cell viability decline. Expression of all the genes studied was up-regulated upon ethanol treatment. In cells treated with 0.5% ethanol, there was a 4.8- (p < 0.001), 6.3- (p<0.001), 5.5- (p<0.05) and 3.6- (p<0.05) fold increase of mRNA levels of IL-6, TNF-α, iNOS and COX-2, respectively, as compared with untreated controls. mRNA levels of the respective genes highly correlated with the applied ethanol concentration. In conclusion, our results show that ethanol treatment may increase the inflammatory potential of AT up-regulating expression of inflammatory cytokines IL-6 and TNF-α and inflammatory enzymes iNOS and COX-2 on transcription level.Scripta Scientifica Medica 2012; 44(2): 31-35

Changes in COX-2, iNOS and GCL gene expression in 3T3-L1 preadipocytes incubated in macrophage conditioned medium

PURPOSE: Obesity is recognized as a major risk factor for a number of diseases, including metabolic syndrome and type 2 diabetes mellitus. It is characterized with increased macrophage infiltration in adipose tissue. Macrophages secrete numerous inflammatory mediators and contribute to the increased inflammatory status of adipose tissue which is associated with increased reactive oxygen species generation and oxidative stress. In cell biology conditioned nutrient mediums (CMs) are used to assess the complex impact of the secretory products of a particular cell type on the behaviour of other cells. The aim of the present study was to examine the effect of CM obtained from J744A.1 macrophage cell culture on expression in 3T3-L1 preadipocytes of some enzymes related to antioxidant defense and the inflammatory response.MATERIAL AND METHODS: mRNA levels of inducible cyclooxygenase (COX-2), inducible NO synthase (iNOS) and glutamate-cysteine-ligase (GCL) were measured. 3T3-L1 cells were treated with increasing CM concentrations (10-30%) for 24 hours. Changes in gene expression levels were analyzed by real-time qPCR method. Calculations were performed using 2-ΔΔCt method.RESULTS: The higher CM concentration contributed to increased expression levels of the genes examined. Treatment with CM in concentrations of 10, 20 and 30% resulted in 2.3-, 2.6- and 2.7-fold increase in COX-2 expression (p<0.01), respectively. Both 10% and 20% CM up-regulated iNOS by 1.7 times (p<0.001). The highest CM concentration of 30% stimulated the enzyme by two times (p<0.01). GCL mRNA levels were by 2.5 times higher than these of the untreated controls when stimulated with 30% CM (p<0.05).CONCLUSION: CM from J744A.1 cell culture may induce inflammatory and oxidative stress-related response in 3T3-L1 preadipocytes thus demonstrating a possible link between macrophage infiltration and local inflammation in adipose tissue.Scripta Scientifica Medica 2013; 45(2): 32-35

Is circulating Gla-rich protein linked with coronary calcium and cardiovascular pathology in patients with atrial fibrillation or heart failure? A pilot study

Introduction: Nowadays Gla-rich protein (GRP) is recognized as a novel biomarker playing a pivotal role in the crosstalk between chronic inflammation and vascular calcification.Aim: The aim of this article is to study the link between circulating GRP, cardiovascular pathology, and the degree of arterial calcification evaluated by the coronary arterial calcium score (CACS) in a Bulgarian population sample. Materials and Methods: Adult participants (n = 81) of both genders were divided into: controls (n = 41)—subjects with estimated moderate-to-high risk without known cardiovascular diseases (CVDs) and a combined CVD group (n = 40)—patients with paroxysmal or persistent atrial fibrillation in sinus rhythm, and heart failure subjects with preserved ejection fraction. A structured interview was carried out for evaluation of the classical CVD risk factors. CACS was determined by multislice computed tomography. Routine laboratory parameters were extracted from medical records. Serum levels of total GRP, matrix Gla protein, and osteocalcin were estimated by commercial ELISA kits. Standard statistical methods (descriptive statistics, Student’s t-test and Spearman’s correlation) were applied. Statistical significance was considered at p<0.05. Results: Significantly lower GRP levels were established in patients with coronary calcium compared to those without calcium deposits. Clear tendency for decreased levels of GRP was observed in the combined CVD group vs controls. Circulating GRP significantly correlates with uncarboxylated matrix Gla protein. An association between serum GRP, CRP, and low-density lipoproteins (LDLs) was demonstrated. Conclusion: This study adds new information regarding the role of circulating GRP as a new player in calcification inhibition. Our findings illuminate the link between total circulating GRP, CVD pathology, and the degree of coronary calcification

Stress-tolerance tests and postprandial low-grade inflammation response

Both tumor necrosis factor alpha (TNF alpha) and interleukin 6 (IL-6) are considered as proinflammatory cytokines which display marked daily and postprandial variations. The low grade inflammation is known to contribute to the development of certain states such as type 2 diabetes, metabolic syndrome and related disorders. Oral glucose tolerance test (OGTT) has been applied to diagnose diabetes and is widely used to estimate the insulin sensitivity in the postprandial phase. Application of tolerance tests with high lipid content such as oral lipid tolerance test (OLTT), high-fat meal (HFM) and mixed meal tests are informative for the metabolic response to complex diets. Postprandial hyperlipidemia and hyperglycemia are important and residual risk factors especially in patients with diabetes mellitus and related metabolic disturbances. There are data suggesting that low grade inflammation could be mediated by hyperglycemia and dyslipidemia. The aim of the review is to summarize how different challenge tests influence the postprandial circulation levels of IL-6 and TNF alpha

Mechanisms of Cyanotoxin Toxicity—Carcinogenicity, Anticancer Potential, and Clinical Toxicology

Cyanoprokaryotes are distributed worldwide and they produce various bioactive compounds, including cyanotoxins. The major route of human exposure to cyanotoxins is the oral intake by using contaminated drinking water, by incidental intake of contaminated water during recreational and professional activities, and by consuming contaminated food or dietary supplements prepared from cyanobacteria. The prolonged chronic exposure to low concentrations of cyanotoxins provokes cell damage and may increase the risk for cancer development. Due to the variety of cyanotoxin chemical structures, different mechanisms of their toxic effects are known. At the same time, some of the natural compounds produced by cyanoprokaryotes have anticancer potential and are promising sources for the development of novel drugs. This chapter is dedicated to the target mechanisms behind the effects of the widely distributed cyanotoxins with an impact on human health, microcystins, nodularins, and cylindrospermopsin

The role of genetic factors and lifestyle relationship to obesity and metabolic syndrome in a sample of Bulgarian population//Проучване ролята на генетични фактори и начина на живот за изявата на затлъстяване и метаболитен синдром в извадка от българската популация

[EN] The dissertation work was dedicated to obesity – serious problem of high medical and social significance of our time widespread all over the world. Certain genetic risk factors for obesity and metabolic syndrome and related biochemical and clinical parameters were studied and evaluated. The allele frequencies of four genes by selected single nucleotide polymorphisms were investigated in a sample of Bulgarian population and compared to the same polymorphisms in different populations. A demographic and social characteristics of the respondents was assessed. A pilot study research for cost-effective and safe approaches for prevention and treatment of obesity and related metabolic disorders was provided. Therefore the anti-inflammatory activity of the medicinal plant Agrimonia eupatoria (agrimony) was studied. On the basis of the studies that scientific work has shown that obesity had polygenic nature. Carriage of a particular genotype may determine predisposition, but social factors determine the obesity phenotype. Higher education level, female sex, younger age and employment were positively correlated with healthy weight. It was found that an aqueous extract of Agrimonia eupatoria had increased antioxidant capacity of blood plasma in humans and had reduced the concentration of certain inflammatory proteins. Today it’s known that obesity is a condition accompanied by oxidative stress, and low-grade chronic inflammation. Hence it might be expected that the medicinal plant agrimony would have a beneficial effect in preventing and treating obesity-related metabolic disorders.[BG] Дисертационният труд е посветен на изключително актуален проблем с висока медицинска и социална значимост на нашето съвремие – затлъстяването. Разгледани са и са оценени някои рискови генетични фактори върху изявата на биохимични и клинични параметри на затлъстяване и метаболитен синдром. Направени са изследвания относно носителство на определен генотип, посредством избрани единични нуклеотидни полиморфизми на извадка от българската популация и са сравнени с носителството на същите полиморфизми в други популации. Направена е демографска и социална характеристика на изследваните лица. В настоящия дисертационен труд е извършено и пилотно проучване в търсене на разход-ефективни и безопасни подходи за фармако и фитотерапия за превенция и лечение на затлъстяване и свързани метаболитни нарушения, като е изследвана противовъзпалителната активност на билката камшик. На основата на извършените проучвания този научен труд показва, че затлъстяването има полигенен характер. Носителството на определен генотип може да определи предразположението, но социо-демографскитре фактори определят степента на затлъстяването. По-високaта степен на образование, женският пол, младата възраст и трудовата заетост корелират положително със здравословното тегло. Установи, че воден извлек от Agrimonia eupatoria L. увеличава антиоксидантния капацитет на кръвна плазма и намалява концентрацията на някои възпалителни фактори. Предвид факта, че затлъстяването е състояние, придружено от оксидативен стрес и нискостепенно хронично възпаление, то би могло да се очаква, че билката ще има благоприятен ефект за превенция и лечение на свързани със затлъстяване метаболитни нарушения

Uric acid effects on glutathione metabolism estimated by induction of glutamate-cysteine ligase, glutathione reductase and glutathione synthetase in mouse J744A.1 macrophage cell line

Introduction: Elevated plasma levels of uric acid (UA) are considered an independent risk factor for hypertension, diabetes, cardiovascular disease, endothelial and vascular damage, obesity, and metabolic syndrome. Even physiological concentrations of soluble UA have been proved to induce gene expression of macrophage-secreted inflammatory cytokines and stimulate production of reactive oxygen species in mature adipocytes. UA is also described as a powerful endogenous plasma antioxidant, which reveals a paradox of duality for this parameter. Aim: The aim of this study was to investigate the effect of UA on expression of antioxidant defense related enzymes in cultured J744A.1 macrophage cell line. Materials and methods: Mouse macrophage J744A.1 cells were treated with uric acid at increasing concentrations of 200 to 800 μM. Changes in expression levels of genes related to the metabolism of glutathione – glutamate-cysteine ligase, catalytic subunit (GCLc), glutathione peroxidase 1 (GPx1), glutathione reductase (GR) and glutathione synthetase (GS) were analyzed. Gene expression levels were calculated using the 2-ΔΔCt method. Results: When UA is applied in concentrations of 200 µM and 400 µM, cell viability did not change significantly. Higher, pathophysiological concentrations of 600 μM, 800 μM UA, and 1000 μM of UA caused significant decrease in cell viability to 95.81% (p<0.01), 76.22% (p<0.001), and 18.01% (p<0.001), respectively. UA treatment in concentrations of 200 μM, 400 μM, 500 μM, and 800 μM induced significant transcription levels of glutathione reductase – 8.14 (p<0.05), 7.15 (p<0.01), 22.07 (p<0.001), and 27.77 (p<0.01), respectively, and of glutathione synthetase – 13.71 (p<0.01), 13.05 (p<0.05), 18 (p<0.01), and 48.60 (p<0.01) folds, respectively. GCLc and GPx1 genes were transcriptionally activated by higher (500 μM and 800 μM) concentrations of UA. For these UA concentrations the measured levels of mRNA were 7.51 (p<0.05) and 12 fold (p<0.05) higher than the non-treated control for GCLc and 1.90 (p<0.05) and 1.93 (p<0.01) for GPx1. Significant difference in the GCLc expression was found between the 200 μM and 500 μM (p<0.05) and 800 μM (p<0.01) treated cells. mRNA levels were significantly different between 400 μM and 800 μM (p<0.05) for both GCLc and GR genes. Very strong correlation was found between GCLc and GR (0.974, p=0.005) and GS (0.935, p=0.020) expression and between GS and GR (0.886, p=0.045) expression levels. Conclusions: It appears that 500 μM and pathophysiological concentrations (800 μM) of UA induce antioxidant cell response in J744A.1 macrophages proved by the indicative elevation GCL, GPx1, GR, and GS transcription. GR and GS can be stimulated even by lower concentrations (200 μM and 400 μM) indicating that glutathione metabolism in macrophages is tightly regulated in order to keep adequate GSH levels

The role of Omega-3 PUFAs in treating the symptoms associated with schizophrenia

Introduction: Schizophrenia is a chronic and severe mental disorder characterized by disconnectiv­ity among distributed brain regions. A schizophrenic has trouble recognizing reality and deception notwithstanding when wakeful. Although schizophrenia is less common than other mental illnesses, the symptoms can be extremely disabling. As polyunsaturated fatty acids (PUFAs) are the major con­stituents of cell membrane phospholipids and take part in the receptors` signalling, they are consid­ered as essential for neurodevelopment and brain functions. The aim of this report is to briefly pres­ent the role of diets rich in omega-3 PUFAs in treating the symptoms associated with schizophrenia.Materials and methods: The presented review is a result of the gathering and analysing of informa­tion from more than 20 articles published in scientific databases - PubMed, Research&Reviews and ScienceDirect.Results: The findings in this study revealed that schizophrenia is a disease that typically begins in early adulthood. Schizophrenic people often have high levels of inflammatory cytokines but low lev­els of omega-3s. The latter play a powerful anti-inflammatory role believed to be responsible for some of their therapeutic effects on mental health. Many randomized controlled clinical trials report that supplementation with omega-3s could lead to significant improvement in symptoms and quality of life. Some of them reported positive effect on primary efficacy, as well as a possibility to prevent full-blown schizophrenia in youngsters having the earliest symptoms. However, other revealed at least some beneficial effect on secondary outcomes, while a single study found no evidence that omega-3s could be a suitable alternative to maintenance antipsychotic treatment in relapse of schizophrenia.Conclusion: Schizophrenia is a multidimensional disorder that prevents people from having a nor­mal life. However, the efficacy of omega-3s is not sufficiently proven yet, hence more scientific data is needed to clarify their role as a promising supplement relieving symptoms of schizophrenia

Changes in interleukin 6 and monocyte chemoattractant protein-1 expression levels in 3t3-l1 cells after exposure to agrimonia eupatoria l. extract and subsequent oxidative stimulation with tert-buthyl hydroperoxide reveal possible adaptogenic potential

INTRODUCTION: Synthesis and secretion of many inflammatory factors increase with oxidative stimu- lation and the activity of many transcription factors that regulate synthesis of inflammatory cytokines is in- fluenced by the redox condition in the  cell.AIM: The aim of the current study was to investigate the effects of aqueous-alcoholic agrimony extract on interleukin 6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) gene expression levels in cultured preadipocytes under induced oxidative stress  conditions.MATERIALS AND METHODS: To determine the possible adaptogenic potential of agrimony extract treatment on gene  expression,  preadipocytes were  incubated  in a  medium containing different  amounts  of agrimony extract. Tert-butyl hydroperoxide was used to provoke oxidative response in treated cells. Ex- pression of selected genes was measured using two step quantitative PCR. Results were analyzed using the 2-ΔΔCt   method.RESULTS: Incubation of preadypocytes with 2.5% agrimony extract resulted in a significant decrease in mRNA levels of MCP-1. Significant increase in IL-6 transcription levels was detected in the cells incubated with 1.25% and 2.5% agrimony extract. Pre-incubation of the cells with agrimony extract prevented subse- quent oxidative induced stimulation of MCP-1 gene expression. Oxidative provocation appeared to decrease the stimulatory activity of agrimony on IL-6 gene  expression