Carbonyl Residues Research Articles

Light-driven molecular motors: imines as four-step or two-step unidirectional rotors.

Chiral N-alkyl imines undergo unidirectional rotation induced by light and heat, thus providing a new class of molecular motors. Depending on the conformational flexibility of the stator part (the carbonyl residue) and the nitrogen inversion barrier of the rotor part (the amine residue) in the molecule, the operation mode of the motor can be controlled as either a four- or a two-step cycling motion of the rotor part.

The anabolic androgenic steroid nandrolone decanoate disrupts redox homeostasis in liver, heart and kidney of male Wistar rats.

The abuse of anabolic androgenic steroids (AAS) may cause side effects in several tissues. Oxidative stress is linked to the pathophysiology of most of these alterations, being involved in fibrosis, cellular proliferation, tumorigenesis, amongst others. Thus, the aim of this study was to determine the impact of supraphysiological doses of nandrolone decanoate (DECA) on the redox balance of liver, heart and kidney. Wistar male rats were treated with intramuscular injections of vehicle or DECA (1 mg.100 g−1 body weight) once a week for 8 weeks. The activity and mRNA levels of NADPH Oxidase (NOX), and the activity of catalase, glutathione peroxidase (GPx) and total superoxide dismutase (SOD), as well as the reduced thiol and carbonyl residue proteins, were measured in liver, heart and kidney. DECA treatment increased NOX activity in heart and liver, but NOX2 mRNA levels were only increased in heart. Liver catalase and SOD activities were decreased in the DECA-treated group, but only catalase activity was decreased in the kidney. No differences were detected in GPx activity. Thiol residues were decreased in the liver and kidney of treated animals in comparison to the control group, while carbonyl residues were increased in the kidney after the treatment. Taken together, our results show that chronically administered DECA is able to disrupt the cellular redox balance, leading to an oxidative stress state.

Oxidative Stress in Chronic Obstructive Pulmonary Disease Alters Ferroxidase Activity of Ceruloplasmin

Aims: Oxidative stress is one of the major patho-physiologic hallmarks in the development of COPD. Ceruloplasmin, the major serum inhibitor of lipid peroxidation has been documented as a main extracellular antioxidant in serum and plays a role in preventing lung injury, and an abnormality in its oxidative inhibition could be involved in pathogenesis of COPD. This study aims to estimate levels of ceruloplasmin and its ferroxidase activity in COPD and compare with that in controls to explore their utility in COPD. Study Design: Comparison study. Place and Duration of Study: Department of Biochemistry, AFMC, Pune. Subjects for COPD were selected from the patients reporting with symptoms of COPD to respiratory OPD of Cardio Thoracic Centre, Pune, during Dec 2010 to Aug 2012. Methodology: Study comprised of two groups: Group I of 77 normal as controls (61 men, 16 women; age range 27-90 years) and Group II of 92 COPD patients (70 men, 22 Original Research Article British Journal of Medicine & Medical Research, 4(5): 1257-1268, 2014 1258 women; age range 45 97 years). Both the groups were further divided into smoker and nonsmoker groups. Ferroxidase activity of ceruloplasmin was estimated by indigenous patented kinetic method of Somani and Ambade while ceruloplasmin was estimated by immunotubidimetric method using commercially available kit. Results: Serum ceruloplasmin and ferroxidase activity were significantly higher in COPD patients as compared to normal controls. Mean ± SD in COPD versus controls respectively are ceruloplasmin: 45.84 ± 12.7 mg/dL versus 37 ± 9.7 mg/dL; ferroxidase: 1324.9 ± 278.53 IU/L versus 980.5 ± 202.3 IU/L, P 0.7) was found between ceruloplasmin and ferroxidase in controls, nonsmoker controls and smoker controls (r = 0.76, 0.71 and 0.79 respectively) while in COPD, COPD nonsmokers and COPD smokers, no correlation was found (r = 0.00, 0.29 and 0.09 respectively). Conclusion: There is alteration in the ferroxidase activity of ceruloplasmin in COPD. Future studies with quantification of carbonyl residues or other groups in ceruloplasmin molecule leading to altered oxidative or ferroxidase activity of ceruloplasmin may provide further evidence to support a role of oxidative stress in COPD.

Macromolecular Oxidation in Planktonic Population and Biofilms of Proteus mirabilis Exposed to Ciprofloxacin

Diverse chemical and physical agents can alter cellular functions associated with the oxidative metabolism, thus stimulating the production of reactive oxygen species (ROS). Proteins and lipids may be important targets of oxidation, and this may alter their functions in planktonic bacterial physiology. However, more research is necessary to determine the precise role of cellular stress and macromolecular oxidation in biofilms. The present study was designed to evaluate whether ciprofloxacin (CIP) could oxidize the lipids to malondialdehyde (MDA) and the proteins to carbonyl residues and to advanced oxidation protein products (AOPP) in planktonic populations and biofilms of Proteus mirabilis. Incubation with CIP generated an increase of lipid and protein oxidation in planktonic cells, with a greater effect found in sensitive strains than resistant ones. Biofilms showed higher basal levels of oxidized macromolecules than planktonic bacteria, but there was no significant enhancement of MDA, carbonyl, or AOPP with antibiotic. The results described in this article show the high basal levels of MDA, carbonyls, and AOPP, with aging and loss of proliferation of biofilms cells. The low response to the oxidative stress generated by CIP in biofilms helps to clarify the resistance to antibiotics of P. mirabilis when adhered to surfaces.

Lipid and protein oxidation products, antioxidant status and vascular complications in poorly controlled type 2 diabetes

The inter-relationships between glycaemic control, the progression of diabetes-related vascular complications, oxidative/antioxidative status and inflammation, have not been fully understood. We measured malondialdehyde (MDA) and carbonyl residues, C-reactive protein (CRP) and antioxidant systems by means of ferric reducing ability of plasma (FRAP) and superoxide dismutase (SOD) activity, in well controlled and poorly controlled type 2 diabetic patients without complications (NC) and in poorly controlled patients with microvascular (MicroVC) and with both micro- and macrovascular complications (Micro+Macro VC). All poorly controlled diabetic patients showed higher MDA and carbonyl residues compared to well controlled NC, as did those with Micro+Macro VC compared to poorly controlled NC. The higher CRP and SOD activity levels reached significance in Micro VC and Micro+Macro VC groups. FRAP decreased only in poorly controlled NC compared to well controlled NC (p<0.05). Glycated hamemoglobin (HbA1c) levels were positively correlated with MDA (p<0.05) and CRP (p<0.001) and inversely associated with FRAP (p<0.05) and SOD (p=0.06). An increase in MDA or carbonyl residues could be a marker of high risk for complications in patients with poorly controlled diabetes and they should be considered for monitoring the effectiveness of drug treatment.

The fatty acid oxidation product 15‐A3t‐Isoprostane is a potent inhibitor of NFκB transcription and macrophage transformation

Fatty acids such as eicosapentaenoic acid (EPA) have been shown to be beneficial for neurological function and human health. It is widely thought that oxidation products of EPA are responsible for biological activity, although the specific EPA peroxidation product(s) which exert these responses have not yet been identified. In this work we provide the first evidence that the synthesized representative cyclopentenone IsoP, 15-A(3t)-IsoP, serves as a potent inhibitor of lipopolysaccharide-stimulated macrophage activation. The anti-inflammatory activities of 15-A(3t)-IsoP were observed in response not only to lipopolysaccharide, but also to tumor necrosis factor alpha and IL-1b stimulation. Subsequently, this response blocked the ability of these compounds to stimulate nuclear factor kappa b (NFκB) activation and production of proinflammatory cytokines. The bioactivity of 15-A(3t)-IsoP was shown to be dependent upon an unsaturated carbonyl residue which transiently adducts to free thiols. Site directed mutagenesis of the redox sensitive C179 site of the Ikappa kinase beta subunit, blocked the biological activity of 15-A(3t)-IsoP and NFκB activation. The vasoprotective potential of 15-A(3t)-IsoP was underscored by the ability of this compound to block oxidized lipid accumulation, a critical step in foam cell transformation and atherosclerotic plaque formation. Taken together, these are the first data identifying the biological activity of a specific product of EPA peroxidation, which is formed in abundance in vivo. The clear mechanism linking 15-A(3t)-IsoP to redox control of NFκB transcription, and the compound's ability to block foam cell transformation suggest that 15-A(3t)-IsoP provides a unique and potent tool to provide vaso- and cytoprotection under conditions of oxidative stress.

Oxidation of mannosyl oligosaccharides by hydroxyl radicals as assessed by electrospray mass spectrometry

The hydroxyl radicals are widely implicated in oxidation of carbohydrates during biological and industrial processes being responsible for their structural modifications and causing functional damage. The identification of intermediate oxidation products is hampered by a lack of reliable sensible methods for their detection. In this study, the oxidation of two models of galactomannans (Man 3 and GalMan 2) has been studied in reaction with hydroxyl radical generated by Fenton reaction. The oxidation patterns were assessed using preparative ligand-exchange/size-exclusion chromatography (LEX/SEC) coupled with tandem electrospray mass spectrometry (ESI-MS/MS). This allowed the identification of derived oligosaccharides (OS) containing hexuronic, hexonic, pentonic and erythronic acid residues and neutral OS bearing hydroperoxy, hydrated carbonyl moieties and residues from pyranosyl ring cleavage. The depolymerization products have been also detected upon oxidation of oligomers. This study allowed developing a simple, effective ‘fingerprinting’ protocol for detecting the damage done to mannans by oxidative radicals.

Comparison of Macromolecular Oxidation by Reactive Oxygen Species in Three Bacterial Genera Exposed to Different Antibiotics

Proteins and lipids maybe important targets of oxidation and this may alter their functions. We evaluated whether ceftazidima (CAZ), piperacillin (PIP), chloramphenicol (CMP), and ciprofloxacin (CIP) could oxidize the macromolecules in the three bacterial genera Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. There was an increase in lipid peroxidation observed in these three species. However, this was lower in the Gram negative bacteria than in S. aureus. A reduction of the carbonyl residue in S. aureus with ciprofloxacin was observed whereas in Gram negative bacteria the antibiotics increased the carbonyl residue with respect to the control. Although the strains suffered a rise in advanced oxidation protein products (AOPP) in the presence of ciprofloxacin, the S. aureus strain had a smaller increase of AOPP than the other strains. The results described in this article provide data about the susceptibility of the three bacterial genera to the oxidative stress induced by the antibiotics studied.

Oxidative modification of albumin in the parenchymal lung tissue of current smokers with chronic obstructive pulmonary disease

BackgroundThere is accumulating evidence that oxidative stress plays an important role in the pathophysiology of chronic obstructive pulmonary disease (COPD). One current hypothesis is that the increased oxidant burden in these patients is not adequately counterbalanced by the lung antioxidant systems.ObjectiveTo determine the levels of oxidised human serum albumin (HSA) in COPD lung explants and the effect of oxidation on HSA degradation using an ex vivo lung explant model.MethodsParenchymal lung tissue was obtained from 38 patients (15F/23M) undergoing lung resection and stratified by smoking history and disease using the GOLD guidelines and the lower limit of normal for FEV1/FVC ratio. Lung tissue was homogenised and analysed by ELISA for total levels of HSA and carbonylated HSA. To determine oxidised HSA degradation lung tissue explants were incubated with either 200 μg/ml HSA or oxidised HSA and supernatants collected at 1, 2, 4, 6, and 24 h and analysed for HSA using ELISA and immunoblot.ResultsWhen stratified by disease, lung tissue from GOLD II (median = 38.2 μg/ml) and GOLD I (median = 48.4 μg/ml) patients had lower levels of HSA compared to patients with normal lung function (median = 71.9 μg/ml, P < 0.05). In addition the number of carbonyl residues, which is a measure of oxidation was elevated in GOLD I and II tissue compared to individuals with normal lung function (P < 0.05). When analysing smoking status current smokers had lower levels of HSA (median = 43.3 μg/ml, P < 0.05) compared to ex smokers (median = 71.9 μg/ml) and non-smokers (median = 71.2 μg/ml) and significantly greater number of carbonyl residues per HSA molecule (P < 0.05). When incubated with either HSA or oxidised HSA lung tissue explants rapidly degraded the oxidised HSA but not unmodified HSA (P < 0.05).ConclusionWe report on a reliable methodology for measuring levels of oxidised HSA in human lung tissue and cell culture supernatant. We propose that differences in the levels of oxidised HSA within lung tissue from COPD patients and current smokers provides further evidence for an oxidant/antioxidant imbalance and has important biological implications for the disease.

Benzyl N-[(S)-2-hy-droxy-1-({[(E)-2-hy-droxy-4-meth-oxy-benzyl-idene]hydrazin-yl}carbon-yl)eth-yl]carbamate.

The shape of the title compound, C19H21N3O6, is curved with the conformation about the imine bond [1.291 (3) Å] being E. While the hy­droxy-substituted benzene ring is almost coplanar with the hydrazinyl residue [N—N—C—C = 177.31 (18)°], an observation correlated with an intra­molecular O—H⋯N hydrogen bond leading to an S(6) ring, the remaining residues exhibit significant twists. The carbonyl residues are directed away from each other as are the amines. This allows for the formation of O—H⋯O and N—H⋯O hydrogen bonds in the crystal, which lead to two-dimensional supra­molecular arrays in the ac plane. Additional stabilization to the layers is afforded by C—H⋯π inter­actions.

Antioxidative mechanisms protect resistant strains of Staphylococcus aureus against ciprofloxacin oxidative damage

The aim of this investigation was to determine whether the antioxidant defences protect resistant strains of Staphylococcus aureus against ciprofloxacin oxidative damage. Reactive oxygen species (ROS) were determined by chemiluminescence and nitric oxide (NO) was assayed by Griess reaction. The accumulation of ciprofloxacin was examined by fluorometry and oxidation of protein, catalase, ferrous reduction antioxidant potency (FRAP), carbonyls and advanced oxidation protein products (AOPP), studied by spectrophotometry. Ciprofloxacin stimulated higher production of ROS and NO in the susceptible strains than in the resistant ones. There was higher accumulation of antibiotic in sensitive strains than in resistant ones, except for the most resistant strain, which accumulated an elevated amount of antibiotic. The FRAP/ciprofloxacin accumulation ratio of the antibiotic was lower in sensitive than in resistant strains. The most resistant strain exhibited the highest FRAP and presented a high catalase activity. There was oxidation of proteins in the presence of ciprofloxacin, with the carbonyl residues increasing in sensitive and resistant S. aureus. The degradation of carbonyls to AOPP in oxidized proteins was higher in the resistant than in sensitive strains. In conclusion, an increase in antioxidant capacity and a rapid oxidation of carbonyls to AOPP contributed to resistance to ciprofloxacin.

Prenylcoumarin with Rev-export inhibitory activity from Cnidii Monnieris Fructus

By use of the fission yeast expressing the model fusion protein comprised of GST, SV40 T antigen NLS, GFP, and Rev-NES in the bioassay, the prenylcoumarin osthol (1) was disclosed as the new Rev-export inhibitor from the MeOH extract of Cnidii Monnieris Fructus. Furthermore, 1 was also found to inhibit export the genuine Rev in HeLa cells by indirect fluorescent antibody technique. By the competitive experiment using the biotinylated probe 3, osthol (1) was revealed to inhibit nuclear export of Rev through a NES non-antagonistic mode. Structure–activity relationship analysis of several analogs of 1 clarified that both prenyl side chain and double bond adjacent to the lactone carbonyl residue play an important role in the Rev-export inhibitory potency of 1.

Bis[2-(ethoxycarbonylamino)ethanaminium] hexabromidostannate

In the title salt, (C5H13N2O2)2[SnBr6], the Sn atom (site symmetry ) exists in a slightly distorted octa­hedral geometry. The cation is non-planar as the terminal CH2NH3 + residue lies below the plane defined by the remaining non-H atoms. In the crystal, cations associate via N—H⋯O hydrogen bonds involving the ammonium and carbonyl residues, forming a 14-membered {⋯HNC2NCO}2 synthon. The cations and anions are arranged in alternating layers arranged along the a-axis direction, the major association between them being N—H⋯Br contacts.

Improvement of functional properties of acid‐precipitated soy protein by the attachment of dextran through Maillard reaction

SummaryThe functional acid‐precipitated soy protein (SAPP)–dextran conjugate was prepared by dry‐heated storage at 60 °C under 79% relative humidity (RH) for 5 days through Maillard reaction between the ε‐amino of lysine in soy proteins and the reducing‐end carbonyl residue in the dextran. The covalent attachment of dextran to SAPP was confirmed by SDS‐polyacrylamide gel electrophoresis and gel filtration chromatography. Functional properties of soy protein depend on the structural and aggregation characteristics of their major components (storage globulins 7S and 11S). The conjugate seemed to be predominantly formed by 7S, and the acidic subunits of 11S in soy protein. The emulsifying properties of the SAPP–dextran conjugate were about four times higher than those of SAPP. The solubility of the protein was not enhanced as a result of preheating, but rather it was not decreased when the conjugated protein was heated at 90 °C for 20 min due to the presence of the polysaccharide. The excellent emulsifying properties of SAPP–dextran conjugate were maintained even at pH 3.0 and were further improved at pH 10.0. The object of Maillard reaction is to guarantee the suitable reaction degree, and the resulting soluble conjugate can have excellent emulsifying properties.

Reactive Immunization Suppresses Advanced Glycation and Mitigates Diabetic Nephropathy

Agents that inhibit glycation end products by reducing the carbonyl load from glycation and glycoxidation are an emerging pharmacologic approach to treat complications of diabetes. We previously demonstrated that antibodies generated to the glycoprotein keyhole limpet hemocyanin (KLH) can cross-link with reactive carbonyl residues on protein conjugates. Here, we immunized streptozotocin-induced diabetic rats with KLH to assess the capacity of the elicited antibodies to intercept carbonyl residues on glycated proteins and to mitigate glycation-related pathology. Compared with diabetic rats immunized with adjuvant alone, KLH-immunized diabetic rats had decreased levels of glycated peptides in sera and demonstrated a reduction in albuminuria, proteinuria, deposition of glycation end products in the kidney, and histologic damage. In vitro, low molecular weight glycated peptides from rat serum reacted with anti-KLH antibodies at a faster rate than normal IgG and selectively modified the lambda chains. The reaction products contained peptide sequences from type I collagen alpha chain, albumin, and LDL receptor-related protein. These adduction reactions were inhibited by free KLH and by reduction of glycated peptides with borohydride. In summary, these results suggest that inherent reactivity of Ig light chains provides a natural mechanism for the removal of cytotoxic glycation products. This reactivity can be augmented by glycoprotein-specific reactive immunization, a potential biopharmaceutical approach to glycation-related pathology.

Lysyl Oxidase Oxidizes Cell Membrane Proteins and Enhances the Chemotactic Response of Vascular Smooth Muscle Cells

Lysyl oxidase (LOX) is a potent chemokine inducing the migration of varied cell types. Here we demonstrate that inhibition of LOX activity by beta-aminopropionitrile (BAPN) in cultured rat aortic smooth muscle cells (SMCs) reduced the chemotactic response and sensitivity of these cells toward LOX and toward PDGF-BB. The chemotactic activity of PDGF-BB was significantly enhanced in the presence of a non-chemotactic concentration of LOX. We considered the possibility that extracellular LOX may oxidize cell surface proteins, including the PDGF receptor-beta (PDGFR-beta), to affect PDGF-BB-induced chemotaxis. Plasma membranes purified from control SMC contained oxidized PDGFR-beta. The oxidation of this receptor and other membrane proteins was largely prevented in cells preincubated with BAPN. Addition of purified LOX to these cells restored the profile of oxidized proteins toward that of control cells. The high affinity and capacity for the binding of PDGF-BB by cells containing oxidized PDGFR-beta was diminished by approximately 2-fold when compared with cells in which oxidation by LOX was prevented by BAPN. Phosphorylated members of the PDGFR-beta-dependent signal transduction pathway, including PDGFR-beta, SHP2, AKT1, and ERK1/ERK2 (p44/42 MAPK), turned over faster in BAPN-treated than in control SMCs. LOX knock-out mouse embryonic fibroblasts mirrored the effect obtained with SMCs treated with BAPN. These novel findings suggest that LOX activity is essential to generate optimal chemotactic sensitivity of cells to chemoattractants by oxidizing specific cell surface proteins, such as PDGFR-beta.

Reactive oxidant species and oxidation of protein and heamoglobin as biomarkers of susceptibility to stress caused by chloramphenicol

Chemiluminescence assay was suitable for rapid determination of alterations caused by chloramphenicol (Ch) in blood. The new aspects observed indicated oxidative stress with: (a) biphasic response of reactive oxidant species (ROS) with different concentration of Ch; (b) increase of carbonyl residues and advanced oxidation protein products in blood as a consequence of oxidative stress; (c) oxidation of haemoglobin with rise of oxyhaemoglobin; protein and haemoglobin oxidation was more pronounced in the samples that exhibited high stimuli of ROS; and (d) the ferrous reduction antioxidant potential (FRAP) was the lowest in the maximum ROS-producer sample, which exhibited the lowest consumption of FRAP in response to Ch treatment. The present investigation could contribute to a rapid detection of persons with elevated susceptibility to Ch previously to its application in chemotherapy.

Nitroxides are more efficient inhibitors of oxidative damage to calf skin collagen than antioxidant vitamins

Reactive oxygen species generated upon UV-A exposure appear to play a major role in dermal connective tissue transformations including degradation of skin collagen. Here we investigate on oxidative damage to collagen achieved by exposure to (i) UV-A irradiation and to (ii) AAPH-derived radicals and on its possible prevention using synthetic and natural antioxidants. Oxidative damage was identified through SDS-PAGE, circular dichroism spectroscopy and quantification of protein carbonyl residues. Collagen (2 mg/ml) exposed to UV-A and to AAPH-derived radicals was degraded in a time- and dose-dependent manner. Upon UV-A exposure, maximum damage was observable at 730 kJ/m 2 UV-A, found to be equivalent to roughly 2 h of sunshine, while exposure to 5 mM AAPH for 2 h at 50 °C lead to maximum collagen degradation. In both cases, dose-dependent protection was achieved by incubation with μM concentrations of nitroxide radicals, where the extent of protection was shown to be dictated by their structural differences whereas the vitamins E and C proved less efficient inhibitors of collagen damage. These results suggest that nitroxide radicals may be able to prevent oxidative injury to dermal tissues in vivo alternatively to commonly used natural antioxidants.

Potassium as an Intrinsic Uncoupler of the Plasma Membrane H+-ATPase

The plant plasma membrane proton pump (H(+)-ATPase) is stimulated by potassium, but it has remained unclear whether potassium is actually transported by the pump or whether it serves other roles. We now show that K(+) is bound to the proton pump at a site involving Asp(617) in the cytoplasmic phosphorylation domain, from where it is unlikely to be transported. Binding of K(+) to this site can induce dephosphorylation of the phosphorylated E(1)P reaction cycle intermediate by a mechanism involving Glu(184) in the conserved TGES motif of the pump actuator domain. Our data identify K(+) as an intrinsic uncoupler of the proton pump and suggest a mechanism for control of the H(+)/ATP coupling ratio. K(+)-induced dephosphorylation of E(1)P may serve regulatory purposes and play a role in negative regulation of the transmembrane electrochemical gradient under cellular conditions where E(1)P is accumulating.

Oxidant and Carbonyl Stress-Related Apoptosis in End-Stage Kidney Disease: Impact of Membrane Flux

Apoptosis is a highly regulated process which mostly affects cell-mediated immunity. In this open-label, randomized, prospective clinical study, we determined the impact in 10 hemodialysis patients treated with high-, medium-, and low-flux membranes on spontaneous or plasma-induced apoptosis, on monocytes, as well as on oxidant and carbonyl stress. High- and medium-flux membranes significantly reduced patients’ plasma-dependent proapoptotic activity on U937 monocytic cell lines. Patients who had the highest levels of plasma-induced proapoptotic activity exhibited the highest plasma levels of advanced oxidation protein products (AOPPs) and carbonyls. Plasma carbonyl residues but not AOPPs were significantly lowered. Finally, a significant correlation could be drawn between the extent of plasma-induced proapoptotic activity and both plasma carbonyl and AOPP levels.