HMGB1/RAGE is identified as a ligand-receptor pair that plays an important role in tumorogenesis. HMGB1 and RAGE levels are higher in most human tumors and their overexpression is associated with tumor progression. The causes of breast cancer are still poorly understood. One reason might be the existence of subtypes within various cellular mechanisms as hormone-dependent and hormone -independent malignant processes. We investigated the effect of HMGB1 protein and its truncated form lacking the C terminus on the RAGE expression and cell motility of breast cancer cell lines; MCF7-noninvasive, MDA-MB-231-invasive and normal breast epithelial one MCF10. The results demonstrate that the effects of HMGB1 and HMGB1∆C through RAGE association are observed exclusively for the hormone independent MDA-MB-231 cell line. The mobility of MDA-MB-231 cells was stimulated only by the full length HMGB1. Our results suggest that HMGB1/RAGE signaling should be considered as an essential process for the development of hormone independent breast cancers with great invasive potential. The truncated form plays the role of a blocking molecule that ”locks” the receptor and inactivates it. This makes the tailless molecule a promising therapeutic agent that competes for the biologically active HMGB1 ligand and prevents the downstream signaling through RAGE.
Thermophilic microbes are present everywhere around us and their only known natural biotope is far away and most usually associated with geothermal energy. To answer this paradox, we explore the hypothesis that the phyllosphere (surface of leaves), due to its exposition to the sun, could well be a thermophilic habitat for microbes and thus a source of thermophilic microbes growing around 50°C – 60°C. To support this hypothesis, we reviewed the heat sources on earth and associated microbial habitats, as well as the difficult identification of thermophilic microbes. We further present an experiment to show the presence and activity of thermophilic bacteria in the phyllosphere. Leaves were collected from eleven tree species from five locations on three continents belonging to three different biomes. On fresh leaves, 16S rDNA sequencing reveals the presence of 0.2 to 7% of clearly identified thermophilic bacteria. Moreover, after incubation at 55°C under aerobic and anaerobic conditions, 16S rDNA sequencing reveals the presence of 4 to 99% of clearly identified thermophilic bacteria. The accumulation of observations provides coherence to our hypothesis and allows the emergence of a new vision of leaves as a thermophilic biotope. We then propose a life cycle of microbes belonging to the thermophilic biotope associated with leaf surfaces.
As an Immune checkpoint blockade therapy (ICB), nivolumab has demonstrated efficacy in Acute Myeloid Leukemia (AML) and various other malignancies. Nivolumab is used as an anti-programmed cell death 1 (PD-1) agent. The toxicities are observed in more than 10% of patients, because of its ability, anti-PD-1 will upregulate the activity of T-cells. Over-activated T-cells will cause immune-related adverse events such as Aplastic Anemia (AA). Here, we present a case of an over 60-years old male patient with AML, and the possibility for him to receive an allogeneic hematopoietic stem cell transplantation (allo-HSCT). The patient was treated with nivolumab and subsequently developed AA. As an additional consideration, we will also discuss whether allo-HSCT is transplantable when AA is performed during the treatment of AML.
Exposure to environmental pollutants can modulate many biological and molecular processes such as gene expression, gene repair mechanisms, hormone production and function and inflammation, resulting in adverse effects on human health including the occurrence and development of different types of cancer. Carcinogenesis is a complex and long process, taking place in multiple stages and is affected by multiple factors. Some environmental molecules are genotoxic, able to damage the DNA or to induce mutations and changes in gene expression acting as initiators of carcinogenesis. Other molecules called xenoestrogens can promote carcinogenesis by their mitogenic effects by possessing estrogenic-like activities and consequently acting as endocrine disruptors causing multiple alterations in cellular signal transduction pathways. In this review, we focus on recent research on environmental chemicals-driven molecular functions in human cancers. For this purpose, we will be discussing the case of two receptors in mediating environmental pollutants effects: the established nuclear receptor, the Aryl hydrocarbon receptor (AhR) and the emerging membrane receptor, G-protein coupled estrogen receptor 1 (GPER1).
Nowadays cardiac simulations are becoming increasingly sophisticated. This trend, part of the maturing field of computational medicine, has provided medical students and cardiologists alike with a new tool for education and research – their very own virtual “patient”. The CircAdapt biophysical model of the human heart and circulation (www.circadapt.org) allows the creation of a virtual “patient” for the study of the cardiovascular system and circulatory haemodynamics under diverse physiological and pathophysiological conditions in real time. The interactive CircAdapt model with its modular design based on established physical and physiologial principles allows dynamic monitoring of blood flow velocities, pressures and volumes in the heart and blood vessels, and across valves and shunts. As an educational tool, the CircAdapt model enables medical students and residents in cardiology, neonatology and intensive care medicine to analyze complex situations while improving their comprehension of cardiovascular physics and (patho)physiology. Moreover, the CircAdapt model has been successfully utilized as a research tool for cardiac resynchronization therapy as well as for various cardiovascular pathologies (e.g. pulmonary arterial hypertension, LBBB). All in all the CircAdapt perspective is as follows: bridge education and research - from classroom to bedside – to foster the future of clinical practice.
Objective: Human Herpes Virus (HHV) type 1 and 2 are cause of hidden pandemics in global scale, as well as sever clinical symptoms associated with active replication in the human host. As until now there are 11 license anti-herpes drugs. Most of them are based on acyclovir and his derivative. Their frequent usage leads to the selection of drug resistance strains and patients offen experience unwanted side effects. Natural products (for ins. plant extracts) are tolerated better by living organisms and their complex composition prevent appearance of resistant virions. The aim of our work is to study the effect of Stachys Thracica Dav extracts against Human Herpes Virus type 1, strain F and Human Herpes Virus type 2, strain BA. Materials and methods: The extracts are obtained from in vivo, in vitro and ex vitro cultivated plants, using methanol extraction. All tests are done in in vitro experimental settings. We use MDBK cell line, and also laboratory strain F of HHV – 1. The following methods were applied: MTT assay to determine cell survival, direct contact assay to test virucidal activity and modified MTT assay to determine effect against virus replication in cell culture. ‡ ‡ ‡ ‡ ‡ ‡,§ § §,‡ © Angelova P et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Results: Obtained data shows that two of the extracts – those from in vitro and ex vitro cultivated plants are with close MNC (maximal nontoxic concentration) (2mg/ml), and the third one is more toxic (MNC is 1mg/ml). The tested extracts do not influence the replication of HHV – 1 and HHV-2. The results about virucidal activity show that the extracts strongly inhibited extracellular virions of HHV-1, strain F (extracts from in vitro cultivated plants reach ~ 100% at 240 minutes of the contact), but the effect of the extracts on extracellular virons of HHV-2, strain BA is more slight. Conclusions: Tested extracts do not have effect against virus replication in cell culture but show strong virucidal activity against HHV-1 and slight activity against HHV-2.
This article aims to explore the antibacterial activity of thin films of TiO2 doped with Ag and Cu using two types of Gram-negative and Gram-positive test bacteria with clinical significance (Gracilicutes and Firmcutes bacteria). The thin films (thickness of about 60 nm) were deposited on glass substrates by radio frequency magnetron co-sputtering (r.f. power of 50 W) of TiO2 target with Ag and Cu pieces on its surface in an Ar atmosphere (0.8 Pa) without heating during the deposition. The total surface area of the Ag was 60 mm2 and that of the Cu was 100 mm2. Bacillus cereus, Staphylococcus epidermidis, Salmonella enterica, Escherichia coli and Pseudomonas sp. were used as test strains. The antibacterial actvity of the films was evaluated by the classical Koch's method and optical density measurements. The bactericidal effect was established at different time points between 30 min and 90 min for Pseudomonas sp. and S. enterica. The Firmicutes bacteria B. cereus and S. epidermidis were killed at the 4th and 8th hour of the treatment, respectively. The effect on E. coli was bacteriostatic until the 10th hour. The results were confirmed by assessment of the bacterial dehydrogenase activity. The studied thin films of TiO2 co-doped with Ag and Cu have a potential for application as antibacterial coatings.
Aim: to study the contribution of structura-functional hemoglobin (Hb) disorders into perinatal hypoxia (PH) pathogenesis and possibility of its indicators using for pharmacotherapy efficiency assessment.90 full-term newborns with PH and 30 healthy ones were examined by clinical methods and Hb spectroscopy. Further children were randomized on 2 groups, received standard therapy and additionally L-carnitine.The indices, indicating Hb affinity to oxygen (1355/1550)/ (1375/1580) and ability to bind them (1355/1550) were less in hypoxia-affected newborns and got reduced at increased severity of PH. The indices of Hb ability to reject ligands (1375/1580) and Hb conformative alterations (1375/1172) were higher in hypoxia-affected neonates and increased simultaneously to PH severity. It means that in severe PH oxygen binding to Hb becomes insufficient. We have established the correlation between Hb conformations and clinical features in newborns with PH. Additional use of L-carnitine promoted restoration of Hb comformative properties and improvement of neurologic status.Our dates confirm the contribution of disorders of Hb structural and functional properties into PH pathogenesis and suggest, that Hb spectroscopy indices could be used as novel criteria of hypoxia severity and pharmaco-therapy efficiency. ‡ § ‡ ‡ ‡ ‡ © Balykova L et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Significant amount of keratinous waste is generated every year, derived from various sources. The structure and complexity of a wool fibre as such, provides resistance and challenge for bacteria to degrade into compounds that can be metabolised. That is why cryogenic pretreatment was applied to improve the digestibility of four sheep breeds fibres. In addition to structural and molecular differences between the untreated, raw wool and treated samples, the effect of pretreatment on methane production for these raw fibres was investigated by anaerobic batch digestion at 37°C. As a response of cryogenic application, soluble protein content had increased in each pretreated sample in comparison with untreated samples, whereas the molecular structure of wool fibres remains intact. The degradation is additionally facilitated by structural changes in the morphological structure. Furthermore, treatment resulted in increased methane production for all treated wool fibres.
Severe burns and scalds almost always result in unsightly hypertrophic scarring. Among the important processes involved in scarring are fibroblast formation and transformation of fibroblasts into myofibroblasts. Myofibroblasts contain α-smooth muscle actin which has contractile properties and can lead to wound contraction and hypertrophic scarring. Phosphorylase kinase (PhK), expressed within 5 mins of injury, is among the earliest enzymes released after tissue damage. It is responsible for activation of NF-kB, which in turn activates over 200 different genes related to inflammation, fibroblastic proliferation, myofibroblast conversion, and eventual scar tissue formation. The sequence and approximate timing of events following injury include the following: activation of PhK (5 mins), followed by appearance of neutrophils (30 mins), macrophages (hours to days), fibroblasts (1 week) and myofibroblasts (2 weeks). Cytokines and growth factors secreted by macrophages include fibroblast growth factor (FGF) and transforming growth factors α and β (TGFα and TGFβ). Fibroblast growth factor is responsible for fibroblastic proliferation, and TGFβ1 for conversion of fibroblasts into myofibroblasts. After thermal injury, the use of topical curcumin, a non-competitive, selective PhK inhibitor that blocks PhK activity upstream of NF-kB activation, was found to be associated with more rapid and improved skin healing, as well as less severe or absent scarring.
Assessment of DNA damage is of primary concern when the pollution-related stress in living organisms has to be determined. The reason for this concern comes from the evidence that damages in DNA often lead to mutations which are potential threat for the sustainability of organisms and ecological systems. To monitor genotoxicity of the marine environment we have developed a special procedure for application of the method of Comet assay (CA) on the native populations of Mytilus galloprovincialis. Samples were collected from areas with different anthropogenic load along the Bulgarian South Black Sea coast. As the first organ to encounter marine pollutants the gills of the mussels turned-out to be the most appropriate tissue for obtaining a single-cell suspension. Comet assay tests were performed and DNA damage was quantified using several different methods including percentage of DNA in the tail, Comet head and tail area, Comet number, Tail moment, Comet shape, etc. The method allowed us to monitor and predict the genetic risk of marine environmental stressors, particularly persistent pollutants. It proved to be convenient for precise quantification of the on-site loads of genotoxic stress on coastal ecosystems.
Electron paramagnetic resonance (EPR) was used as a method for recording the content of the nitric oxide (NO) in hippocampal tissues of intact rats and rats after modelling of ischaemic and haemorrhagic stroke. Based on direct measurements of NO by EPR spectroscopy, it was shown that, within 5 hours after the onset of symptoms of ischaemic and haemorrhagic stroke, the formation of NO in the hippocampus was reduced by a factor of 2-3 and this reduction was maintained for a period of between 24 and 72 hours. The results show that a systemic character of a decrease in the intensity of NO production during the modelling of ischaemic events in the brain reflects the effects of central dysregulation of the functions at the level of the whole organism such that it is appropriate to consider implementing the correction of the vital systems of the body in a stroke. It has indicated that non-selective NO-synthase blocker L-NAME reduced the low level of NO production by a factor of 3 by its administration within 72 hours after post-ischaemic and haemorrhagic stroke. It was discovered however that L-NAME returns the level of NO production to baseline (control) by its administration within 5 hours after ischaemia.
Effective therapies for multiple sclerosis (MS) are still missing. This neurological disease affects more than 2.5 million people worldwide. To date, biological immunomodulatory drugs are effective and safe during short-term treatment, but they are suitable only for parenteral administration and they are expensive. Accordingly, academic and industrial environments are still focusing their efforts toward the development of new MS drugs. Considering that neurodegeneration is a contributory factor in the onset of MS, herein we will focus on the crucial role played by sigma 1 receptors (S1Rs) in MS. A pilot study was performed, evaluating the effect of the S1R agonist (R)-RC33 on rat dorsal root ganglia experimental model. The encouraging results support the potential of S1R agonists for MS treatment.
In this study, we present the results of the preliminary screening of a library of microorganisms, collected from different environmental settings. Approximately 300 strains of the culture collection were tested on solid medium for inhibition of growth of three tester species, namely Bacillus subtilis, Escherichia coli and Staphylococcus aureus. The selection of the tester species was made according to the following criteria: Staphylococcus aureus and Escherichia coli were chosen as representative of Gram+ and Gram- bacteria, respectively. Furthermore, these organisms are relevant for the global public health because the number of antibiotic resistant strains responsible for invasive diseases is steadily increasing (Centers for Disease Control and Prevention 2014, Lowy 2003); Bacillus subtilis, instead, was selected as Gram+ tester microorganism commonly found in soil samples. Thus, this well characterized bacterium can be used to gain insight on the effect of metabolites produced by microorganisms of the culture collection described in this study. One of the active strains, MES18, was classified as a Bacillus spp. by means of small-subunit rRNA gene sequencing. To identify the compound(s) responsible for this inhibitory activity, MES18 cells were grown in liquid medium at 30°C and samples were taken at different time points over a period of 12 days. The supernatants obtained from the fermentation media were subjected to fractionation by chromatography on reversed-phase column and all the eluted compounds were assayed for their ability to repress the growth of tester strains. This approach allowed us to identify the fractions containing the bioactive compound(s) and to establish that the production of these secondary metabolites reached a maximum during the idiophase of the cell culture, when cell growth and replication decline. Further analyses to identify the physical-chemical features of the compound(s) produced by this strain using HPLC coupled to mass-spectrometry are currently ongoing.
During the last two decades, perivascular adipose tissue (PVAT) has been revealed as an important regulator of vascular processes such as proliferation of smooth muscle cells, proand anti-oxidant reactions in the vascular wall, angiogenesis, inflammation, apoptosis of neutrophils, migration of monocytes and others.PVAT derived mediators either increase or decrease the amplitudes of the force of artery contraction measured using isometric small vessel myography.In healthy animals and humans predominates the relaxing effect while in diseases the contractile influence of PVAT is common.In aging and pathological conditions like atherosclerosis and diabetes, or with environmental factors like tobacco smoke and high-fat diet, the phenotype of perivascular adipocytes is changed from antiinflammatory to pro-inflammatory.This change is accompanied by a significant rearrangement of mediators released from PVAT.
There are some data about genes associated with certain traits of a person in the scientific literature, but these data are often contradictory. It is evident that complexes of genes that affect the manifestation of the trait can be more informative. We have investigated genotypes of 9000 people, using the PCR method, in order to determine their athletic abilities or predisposition to different diseases and pathologies. In particular, we tested about 3000 women with unknown causes of miscarriages for 14 genes associated with the pregnancy development. 1,5 years later we interviewed 700 women and have identified a high genetic risk of pregnancy loss due to hereditary thrombophilia. In most of the cases, doctors took into account our data and applied for these women treatment with anticoagulant drugs such as fragmin in the subsequent pregnancies. So 86.6% of pregnant women have successfully conceived and given birth, and we have received over 500 touching letters of gratitude. Thus, genetic testing allows to identify individual human properties in order to prevent some pathologies or to chose the most suitable kind of sport.
Osteoporosis causes bones to become weak and brittle. It is known that the alterations in bone metabolism associated to osteoporosis can impair bone healing around implants and affect their osseointegration. The main objective of this study was the development of new nanostructured implant materials based on ultrananocrystalline diamond (UNCD) coatings for enhancing osseointegration. The films were deposited on Ti substrates by microwave plasma CVD from 17% CH4/N2 gas mixtures and modified by O2 or NH3/N2 plasmas. The modifications rendered the H-terminated hydrophobic as-grown films hydrophilic. The interaction of endothelial (EA.hy926) and osteosarcoma (MG63) cells with differently modified UNCD surfaces was investigated by proteome analyses. It revealed the identification of over 19 000 proteins (extracellular and cytosolic). They correspond to 508 (Ti), 634 (UNCD), 651 (O-UNCD), and 668 (NH2-UNCD) protein groups. The interaction network analysis showed differences in the connectivity of inferred protein networks between the ECM niches, which suggests the presence of specific cell microenvironments on O- and NH2-terminated UNCD surfaces. Our results show that due to a favorable combination of surface chemical and topological properties the UNCD films, as-grown and especially after their plasma modifications, may serve as superior scaffolding for promoting the cell attachment and growth during osseointegration.
Thymidine kinase (TK) has been validated as a serum-derived, tumour-associated marker for a number of malignancies and estimation of TK activity in serum has proved useful for clinical diagnosis and monitoring of therapy. However, the use of this biomarker in the clinical practice is constrained by the lack of an automatable easy-to-perform assay. We have developed and validated a novel HPLC-based assay for measuring TK activity in biological samples. This assay is cheaper, easy to perform and does not depend on the use of expensive antibodies or isotopes. In addition, it has comparable sensitivity with the radioenzymatic assay used in the clinical practice. The assay has been evaluated with samples from breast cancer patients.
