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BACKGROUND: Combining pathogen reduction and automated separation of whole blood (WB), together with the use of improved additive solutions, may increase reproducibility and extend shelf-life of blood components. MATERIALS AND METHODS: Forty WB units were collected from volunteer donors and randomised 1:1 into two groups: 1) pathogen reduction with riboflavin and ultraviolet light (PRT); or 2) no treatment (Control). After two hours (h) at room temperature, all units underwent fully automated separation into red blood cell concentrate (RBCC), plasma and leukopack components. RBCCs were leukoreduced and stored in phosphate-adenine-glucose-guanosine-saline-mannitol (PAGGSM) solution while plasma units were shock frozen within 8 h of collection and stored at ≤ −25°C. RBCCs were sampled on day 1 and weekly thereafter until day 42, while plasma was sampled on days 1 and 30. The main study objective was to assess the in vitro quality of separated RBCCs using biochemical and haematological parameters. Plasma protein content after one cycle of freeze-thaw was also analysed. RESULTS: The quality of RBCCs was largely comparable between the PRT and Control groups, except for a significantly higher degree of haemolysis and extracellular potassium levels in the PRT group after 35 days of storage. While potassium concentration was significantly higher in the PRT group at all timepoints, the degree of haemolysis exceeded the accepted European threshold (i.e.

The aim of this study was to evaluate the nature and mechanisms of interaction between HIV peptide/dendrimer complexes (dendriplex) and artificial lipid membranes, such as large unilayered vesicles (LUV) and lipid monolayers in the air–water interface. Dendriplexes were combined as one of three HIV-derived peptides (Gp160, P24 and Nef) and one of two cationic phosphorus dendrimers (CPD-G3 and CPD-G4). LUVs were formed of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) or of a mixture of DMPC and dipalmitoyl-phosphatidylglycerol (DPPG). Interactions between dendriplexes and vesicles were characterized by dynamic light scattering (DLS), fluorescence anisotropy, differential scanning calorimetry (DSC) and Langmuir–Blodgett methods. The morphology of formed systems was examined by transmission electron microscopy (TEM). The results suggest that dendriplexes interact with both hydrophobic and hydrophilic regions of lipid bilayers. The interactions between dendriplexes and negatively charged lipids (DMPC–DPPG) were stronger than those between dendriplexes and liposomes composed of zwitterionic lipids (DMPC). The former were primarily of electrostatic nature due to the positive charge of dendriplexes and the negative charge of the membrane, whereas the latter can be attributed to disturbances in the hydrophobic domain of the membrane. Obtained results provide new information about mechanisms of interaction between lipid membranes and nanocomplexes formed with HIV-derived peptides and phosphorus dendrimers. These data could be important for the choosing the appropriate antigen delivery vehicle in the new vaccines against HIV infection.

Radiotherapy is among the most important methods for breast cancer treatment. However, this method’s effectiveness is limited by radioresistance. The aim of this study was to investigate whether the stilbene derivatives piceid, resveratrol, and piceatannol have a radiosensitising effect on breast cancer cells (MCF-7). The conducted research enabled us to determine which of the tested compounds has the greatest potential in sensitising cells to ionising radiation (IR). Among the stilbene derivatives, resveratrol significantly increased the effect of IR. Resveratrol and IR used in combination had a higher cytotoxic effect on MCF-7 cells than using piceatannol, piceid, or radiation alone. This was due to a significant decrease in the activity of antioxidant enzymes, which resulted in the accumulation of formed reactive oxygen species (ROS). The effect of resveratrol and IR enhanced the expression of apoptotic genes, such as Bax, p53, and caspase 8, leading to apoptosis.

Purpose: This work investigates the effect of resveratrol and melatonin on structural and functional changes of two enzymes, lactate dehydrogenase (LDH) and glyceraldehyde-3-phosphate dehydrogenase...

There is increasing evidence that supplementation with superfoods is one of the available solution to prevent or slow down the progression of neurodegenerative diseases. Especially polyphenols have been reported to exert strong neuroprotective properties. Taking into consideration our latest discovery, where we demonstrated that piceatannol inhibits glyceraldehyde-3-phosphate dehydrogenase (GAPDH) aggregation (in tube) we decided to explore its effect on hippocampal cells. We investigated how piceatannol influence on oxidatively modified GAPDH in hippocampal cells by using confocal microscopy, flow cytometry, western blot analysis, spectrophotometric assays. Additional we assessed effect of piceatannol on level of Bdnf and Sirt1 mRNA (RT-PCR) and the activity of GPx, GR, GST, Cat. We found that piceatannol significantly suppresses GAPDH nuclear translocation as well as protein aggregation induced by excessive stress. The piceatannol anti-aggregation activity and ability to counteract GAPDH nuclear translocation place this compound as a new drug candidate for in vivo tests.

A number of independent studies have shown the contribution of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the pathogenesis of several neurodegenerative disorders. Indeed, GAPDH aggregates have been found in many post-mortem samples of brains of patients diagnosed with Alzheimer's and Parkinson disease. Currently, it is accepted that GAPDH-mediated cell death pathways in the neurodegenerative processes are associated with apoptosis caused by GAPDH nuclear translocation and excessive aggregation under oxidative stress conditions. Also the role of GAPDH in neurodegenerative diseases is linked to it directly binding to specific amyloidogenic proteins and petides such as β-amyloid precursor protein, β-amyloid peptide and tau protein in Alzheimer's disease, huntingtin in Huntington's disease and α-synuclein in Parkinson disease. One of the latest studies indicated that GAPDH aggregates significantly accelerate amyloidogenesis of the β-amyloid peptide, which implies that aggregates of GAPDH may act as a specific aggregation "seed" in vitro. Previous detailed studies revealed that the active-site cysteine (Cys152) of GAPDH plays an essential role in the oxidative stress-induced aggregation of GAPDH associated with cell death. Furthermore, oxidative modification of this cysteine residue initiates the translocation of the enzyme to the nucleus, subsequently leading to apoptosis. The crystallographic structure of GAPDH shows that the Cys152 residue is located close to the surface of the molecule in a hydrophilic environment, which means that it can react with low molecular weight compounds such as hydroxynonenal or piceatannol. Therefore, it is highly possible that GAPDH may serve as a target for small molecule compounds with the potential to slow down or prevent the progression of neurodegenerative disorders. Recently appearing new evidence has highlighted the significance of low molecular weight compounds in counteracting the oxidation of GAPDH and consequently its aggregation and other unfavourable pathological processes. Hence, this review aims to present all recent findings concerning molecules that are able to interact with GAPDH and counteract its aggregation and translocation to the nucleus.

Background Nitric oxide is a well-known gaseous signaling molecule and protein modifying agent. However, at higher concentrations or during oxidative stress nitric oxide may exert some deleterious effects on protein structure and function. Here we investigated the influence of nitric oxide and products of its oxidation on two glycolytic enzymes: GAPDH and LDH under in vitro nitrosative stress conditions. Secondly, we applied natural antioxidants: melatonin and resveratrol to examine their effects on the enzymes under studied conditions. Methods By means of UV–VIS and fluorescence spectroscopy methods we compared nitric oxide mediated changes of enzyme activities, amount of free sulfhydryl groups (-SH) and bis-ANS probe binding. Finally, we predicted potential cysteine residues modified by nitric oxide in studied proteins using GPS-SNO software. Results Our results indicated that nitric oxide reversibly inactivates GAPDH but does not affect the activity of LDH. Nitric oxide dependent GAPDH activity decline was accompanied by the reduction of the amount of free –SH groups and GAPDH-bound bis-ANS fluorescence. Reduction of the number of free –SH groups and protein-bound bis-ANS fluorescence was also observed in LDH treated with NO. Applied antioxidants increased inactivation of GAPDH and structural changes of GAPDH and LDH. Conclusions Nitric oxide modifies function and structure of thiol-dependent enzyme such as GAPDH and structure of LDH which function do not rely on cysteine thiols. Both resveratrol and melatonin exerted prooxidative properties in studied conditions. General significance Extensively studied antioxidants: resveratrol and melatonin may function as a prooxidative species under in vitro nitrosative stress conditions.

The pathogenesis of neurodegenerative diseases involves oxidative stress-induced cell death. It’s well documented that polyphenolic compounds protect neurons against this type of cell damage. Among many polyphenols, stilbenoids derivatives are of particular interest given their potential for use in therapeutics and prevention of neurodegenerative disorders. Piceatannol, which is ubiquitous in variety of edible plants has broad spectrum of action including antioxidant, anti-inflammatory, anti-cancer, and neuroprotective activities. In the present study, the effects of piceatannol against oxidative stress-induced cell damage in neuroblastoma cells were evaluated. Our data indicate that protective effect of piceatannol in neuroblastoma cells under oxidative stress conditions is associated with increased activity and expression of glutathione peroxidase enzymes. Piceatannol also stimulates BDNF, which is involved in neurite outgrowth and maintenance of function in neuronal cells. Furthermore, piceatannol promotes cell survival by sustaining level of sirtuin 1 and seladin-1 mRNA on constant level.

The history of using performance-enhancing substances (PES) is long and it goes back to ancient times. At present, PES are employed at all levels of sport competition, starting from Olympic level contestants to individuals recreationally involved in various sports disciplines.e objective of the study was examining the views on doping in sports in a group of physicians, together with evaluating the frequency of their contacts with this phenomenon, in their professional activities.e investigation was carried out using a validated questionnaire developed by the authors. Questionnaire included 34 questions divided into 6 sections. In total, 257 individuals participated in the study. e percentage of answers was 75.81%.Among the responders, 96.50% believed that using PES by sports competitors represented unethical behavior. 42% participants declared that they met doping problem during their professional career. Almost one-third of the physicians (28.79%) declared that during their work, they consulted patient suffering from adverse side effects resulting from using PES.In Poland, physicians regard using PES as unethical behavior. They believe that a health care system professional should participate in counteracting doping in sports. Physicians - regardless of their specialty - are also exposed to PES-related problems in their professional work. In view of the above facts and the consistently increasing popularity of PES, extending the knowledge in this field among physicians seems to be of importance to allow for their offering better medical services to their patients.

Proteins, which have enzymatic activities play a fundamental role in the cell due to participation in most of biological processes. Oxidative-induced damage of enzymes often have marked effects on cellular processes, which in consequence determine cell functioning and survival. In this review, we focused on the radiation-induced inactivation of enzymes with particular emphasis on the inactivation of dehydrogenases. For a better understanding of this issue, the efficiency of products of water radiolysis (∙OH, O 2 ∙ − and H 2 O 2 ) in enzyme inactivation has been analysed. Reactions of reactive oxygen species (ROS) with amino acids present in the active site of enzymes appear to have the greatest impact on enzyme inactivation.

Background Overactive bladder syndrome is a condition where one or more of the symptoms such as pollakiuria, urgent need to urinate, nocturia and urinary incontinence is observed. Its prevalence ranges between 7 and 27 % in men and 9–43 % in women. The role of a pharmacist is to educate the patient on medications administration scheme, and drug interactions with particular food or food components. Aim of the review To assess a potential impact of food and fruit juice on the pharmacokinetic and therapeutic effects of medications used in treating overactive bladder syndrome. This information will enhance pharmaceutical care and is vital and helpful for pharmacists counseling their patients. Method In order to gather information on interactions of medications employed in bladder dysfunctions, the English language reports published in the PubMed, Embase, Cochrane and CINAHL database over the years 1996–2015 were studied. Additionally, other resources, namely drugs.com, Medscape, UpToDate, Micromedex, Medical Letter, as well as Stockley Drugs Interaction electronic publication were included in the study. The analysis also covered product data sheets for particular medicinal products. Results Meals and the consumption of grapefruit juice were found to exert a diversified effect on the pharmacokinetics of drugs employed in overactive bladder syndrome therapy. Neither tolterodine, nor mirabegron interact with food and citrus fruit juice, whereas darifenacin, fesoterodine, oxybutynin and solifenacin do interact with grapefruit and others citrus fruit juice. The effects of such interactions may potentially be negative to patients. Trospium absorption is significantly decreased by food. Conclusion For selected medicines used in treating bladder dysfunctions food and grapefruit juice consumption may significantly affect efficacy and safety of the therapy. All information on the topic is likely to enhance the quality of pharmaceutical care.

World Health Organization (WHO) defines adverse drug reaction (ADR) as “a response to a drug which is noxious and unintended, and which occurs at doses normally used in man for the prophylaxis, diagnosis, or therapy of disease, or for the modifications of physiological function”. ADRs are a serious problem of contemporary pharmacotherapy. Expenditures for treatment of ADRs in the United States may cost up to 30.1 billion dollars annually. Factors affecting the development of ADRs are: age, gender, body weight, polypharmacy. About 10% of ADRs is associated with gastrointestinal tract (GIT). ADR can affect every part of GIT. Xerostomia is the most common ADR occurring in oral cavity. ADRs affecting esophagus include irritation and inflammation of the mucosa. Approximately one-third of all cases of esophageal inflammation results from administration of non-steroid anti-inflammatory drugs (NSAIDs). The main cause of ulcerations involving stomach and small intestine are NSAIDs. Drug-induced diarrheas are the most common adverse effect accounting for approximately 7% of all observed cases of ADRs. They may be triggered by antibiotics, magnesium salts, laxatives and others. On the other hand, some groups of medications may induce constipation. These drugs comprise opioids, diuretics, calcium channel blockers, cholinolytics and others. Proton pump inhibitors, metformin, orlistat and colesevelam may lead to restricted absorption of certain vitamins and minerals. Physicians’ knowledge about most popular and well documented ADRs can improve patients’ safety and make pharmacotherapy more comfortable for them.

Introduction Increased oxidative stress has been implicated in the pathogenesis of Crohn disease (CD). Except for C‑reactive protein (CRP), good biological markers of CD activity are lacking. Objectives We aimed to investigate the diagnostic usefulness of selected markers of oxidative stress in the serum and saliva of patients with active and inactive CD. Patients and methods A total of 58 patients with confirmed CD (32 with active CD, 26 with inactive CD, and 26 healthy controls) were prospectively enrolled to the study. The markers examined were malondialdehyde (MDA), ferric reducing ability of plasma (FRAP), reduced glutathione (GSH), and catalase (CAT). Results MDA levels were higher in the serum and saliva of patients with active CD than in those with inactive CD and controls and were positively correlated with the Crohn's Disease Activity Index (r = 0.8, P

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is one of the key redox-sensitive proteins whose activity is largely affected by oxidative modifications at its highly reactive cysteine residue in the enzyme’s active site (Cys149). Prolonged exposure to oxidative stress may cause, inter alia, the formation of intermolecular disulfide bonds leading to accumulation of GAPDH aggregates and ultimately to cell death. Recently these anomalies have been linked with the pathogenesis of Alzheimer’s disease. Novel evidences indicate that low molecular compounds may be effective inhibitors potentially preventing the GAPDH translocation to the nucleus, and inhibiting or slowing down its aggregation and oligomerization. Therefore, we decided to establish the ability of naturally occurring compound, piceatannol, to interact with GAPDH and to reveal its effect on functional properties and selected parameters of the dehydrogenase structure. The obtained data revealed that piceatannol binds to GAPDH. The ITC analysis indicated that one molecule of the tetrameric enzyme may bind up to 8 molecules of polyphenol (7.3 ± 0.9). Potential binding sites of piceatannol to the GAPDH molecule were analyzed using the Ligand Fit algorithm. Conducted analysis detected 11 ligand binding positions. We indicated that piceatannol decreases GAPDH activity. Detailed analysis allowed us to presume that this effect is due to piceatannol ability to assemble a covalent binding with nucleophilic cysteine residue (Cys149) which is directly involved in the catalytic reaction. Consequently, our studies strongly indicate that piceatannol would be an exceptional inhibitor thanks to its ability to break the aforementioned pathologic disulfide linkage, and therefore to inhibit GAPDH aggregation. We demonstrated that by binding with GAPDH piceatannol blocks cysteine residue and counteracts its oxidative modifications, that induce oligomerization and GAPDH aggregation. This work is supported by a grant from the Faculty of Biology and Environmental Protection, University of Lodz (grant number: B1711000001504.02) and by National Science Centre, Poland (grant number 2017/25/N/NZ1/02849) and supported by subsidy for young scientists (Faculty of Biology and Environmental Protection, University of Lodz). Bionanopark Ltd. is a non-profit research institution providing employment for two of the authors [JG, JAB], whose facilities were used for part of the conducted studies. The funder provided support in the form of research materials, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

To determine the mechanism underlying oxidative modifications caused by radiation-induced reactive oxygen species (ROS) and elucidate their effect on the structure and function of yeast alcohol dehydrogenase (YADH), a zinc-containing protein.YADH was exposed to water radiolysis products in an air atmosphere. YADH oxidation products were determined by spectrophotometric and spectrofluorimetric methods. The extent to which oxidative modifications affected enzyme activity was also studied.Water radiolysis products oxidized thiol groups leading to the release of zinc ions and the destruction of tryptophan and tyrosine residues. Those processes were accompanied by alterations in protein structure such as increased surface hydrophobicity, greater tryptophan accessibility to acrylamide, and changes in the secondary structure. Structural modifications were correlated with lower enzyme activity.During the process of functional and structural changes in YADH exposed to reactive oxygen species, a key part is the oxidation of cysteine residues attached to zinc and the release of zinc ions from the molecule. It may be assumed that ROS induce similar changes in many other zinc-containing proteins.

The presented study analyzed potential binding sites of 3,5,4'-trihydroxystilbene (resveratrol, RSV) and its derivative, trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene (THMS) to glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The effects of stilbene analogs on the structure of GAPDH were determined by fluorescence spectroscopy and ζ potential measurements. To what extent the studied compounds affect the activity of the enzyme was also assessed. A computational ligand-docking study showed that there are 11 potential binding sites of RSV and 8 such sites of THMS in the GAPDH molecule. While resveratrol does not significantly affect the activity of the dehydrogenase upon binding to it, THMS leads to approximately 10% inactivation of this enzyme. THMS has no effect on GAPDH inactivation induced by the superoxide anion radical, in contrast to resveratrol, which increases dehydrogenase inactivation.

INTRODUCTION Insulin‑like growth factor 1 (IGF‑1) is involved in the modulation of immunity and inflammation. It also plays a role in regulating the migration of endothelial cells and production of vasoactive agents. OBJECTIVES This study assessed the concentrations of IGF‑1 and insulin‑like growth factor-binding protein 3 (IGFBP‑3) and their relationships to disease activity in patients with inflammatory bowel disease (IBD). PATIENTS AND METHODS A total of 129 adult patients with IBD (69 with Crohn disease [CD] and 60 with ulcerative colitis [UC]) were involved in the study. The control group consisted of 31 healthy volunteers. Biochemical serum analyses were performed and the associations of IGF‑1 and IGFBP‑3 with inflammatory markers and disease activity were assessed. RESULTS IGF‑1 levels were decreased in patients with active UC compared with those with nonactive UC (mean [SD], 78.3 [22.7] ng/ml and 96.2 [24.5] ng/ml, respectively; P = 0.02) and controls (94.5 [26.5] ng/ml; P = 0.03). The IGF‑1 level was lower in patients with active CD compared with those with nonactive CD (mean [SD], 79.2 [24.9] ng/ml and 110.1 [43.4] ng/ml, respectively; P

In the present study the effect of X-irradiation on the alcohol dehydrogenase (ADH) activity in the presence of nanoparticles of fullerenol C 60 (OH) ~30 under aerobic conditions was investigated in order to assess the potential radioprotective properties of fullerenol. Fullerenol at 75 mg/mL decreased the radiation yield of inactivation of ADH irradiated with fullerenol by 20% comparing to ADH irradiated without fullerenol. Under conditions used during irradiation, 50% of OH radicals could react with fullerenol and 50% could react with ADH. Thus, it can be assumed that protective effect of fullerenol on the radiation inactivation of ADH was mostly due to scavenging OH radicals by fullerenol. Moreover, fullerenol did not protect against post-irradiation damage as the G inact for ADH irradiated with fullerenol was still 20% lower than for ADH irradiated without fullerenol after 24 h from irradiation. Additionally, fullerenol at 75 mg/L had no influence on the activity of unirradiated ADH up to 24 h. We concluded that fullerenol C 60 (OH) ~30 protected ADH against radiation inactivation due to simple competition for the OH radicals and did not modify its activity by association with the protein as it was proved in our previous papers for erythrocyte membrane proteins.