Thinking beyond Vaccination: Promising Add-On Strategies to Active Immunization and Vaccination in Pandemics-A Mini-Review.

There is little doubt that final victories over pandemics, such as COVID-19, are attributed to herd immunity, either through post-disease convalescence or active immunization of a high percentage of the world's population with vaccines, which demonstrate protection from infection and transmission and are available in large quantities at reasonable prices. However, it is assumable that humans with immune defects or immune suppression, e.g., as a consequence of allograft transplantation, cannot be immunized actively nor produce sufficient immune responses to prevent SARS-CoV-2 infections. These subjects desperately need other strategies, such as sophisticated protection measures and passive immunization. Hypertonic salt solutions attack vulnerable core areas of viruses; i.e., salt denatures surface proteins and thus prohibits virus penetration of somatic cells. It has to be ensured that somatic proteins are not affected by denaturation regarding this unspecific virus protection. Impregnating filtering facepieces with hypertonic salt solutions is a straightforward way to inactivate viruses and other potential pathogens. As a result of the contact of salt crystals on the filtering facepiece, these pathogens become denatured and inactivated almost quantitatively. Such a strategy could be easily applied to fight against the COVID-19 pandemic and other ones that may occur in the future. Another possible tool to fight the COVID-19 pandemic is passive immunization with antibodies against SARS-CoV-2, preferably from human origin. Such antibodies can be harvested from human patients' sera who have successfully survived their SARS-CoV-2 infection. The disadvantage of a rapid decrease in the immunoglobulin titer after the infection ends can be overcome by immortalizing antibody-producing B cells via fusion with, e.g., mouse myeloma cells. The resulting monoclonal antibodies are then of human origin and available in, at least theoretically, unlimited amounts. Finally, dry blood spots are a valuable tool for surveilling a population's immunity. The add-on strategies were selected as examples for immediate, medium and long-term assistance and therefore did not raise any claim to completeness.

Lipidomics Revealed Plasma Phospholipid Profile Differences between Deceased and Recovered COVID-19 Patients.

Thorough understanding of metabolic changes, including lipidome alteration, associated with the development of COVID-19 appears to be crucial, as new types of coronaviruses are still reported. In this study, we analyzed the differences in the plasma phospholipid profiles of the deceased COVID-19 patients, those who recovered and healthy people. Due to identified abnormalities in plasma phospholipid profiles, deceased patients were further divided into two subgroups (D1 and D2). Increased levels of phosphatidylethanolamines (PE), phosphatidylcholines (PC) and phosphatidylserines (PS) were found in the plasma of recovered patients and the majority of deceased patients (first subgroup D1) compared to the control group. However, abundances of all relevant PE, PC and PS species decreased dramatically in the plasma of the second subgroup (D2) of five deceased patients. These patients also had significantly decreased plasma COX-2 activity when compared to the control, in contrast to unchanged and increased COX-2 activity in the plasma of the other deceased patients and recovered patients, respectively. Moreover, these five deceased patients were characterized by abnormally low CRP levels and tremendous increase in LDH levels, which may be the result of other pathophysiological disorders, including disorders of the immune system, liver damage and haemolytic anemia. In addition, an observed trend to decrease the autoantibodies against oxidative modifications of low-density lipoprotein (oLAb) titer in all, especially in deceased patients, indicate systemic oxidative stress and altered immune system that may have prognostic value in COVID-19.

Occupational Health Aspects with Special Focus on Physiological Differences between Office and Metalworkers.

Physical workload adversely impacts inflammation, oxidative stress and mood in heavy workers. We compared these risk parameters between metalworkers (n = 20) and office workers (n = 30), including gender differences. Blood samples were analyzed with thirty parameters to overview endocrinology, inflammation, and psychological and oxidative stress. Despite an adequate antioxidative supply, oxidative stress occurred in metalworkers, as indicated by significantly increased peroxide and homocysteine (Hcy) levels. Moreover, increased concentrations were observed in this group regarding psychological stress and diet-related parameters. Sex-specific differences were determined for physical dimensions, dehydroepiandrosterone sulfate (DHEAS), Hcy, uric acid, triglycerides, osmolality, anti-Mullerian hormone (AMH) and testosterone. Age-associated differences were observed for DHEAS, glycosylated hemoglobin, adrenaline, AMH and testosterone. In male office workers, the body mass index was associated with increased LDL-HDL, cholesterol-HDL and homeostatic model assessment of insulin resistance (HOMA-IR). In conclusion, these results indicate increased oxidative stress and psychological stress in heavy workers independently of adequate antioxidant sustenance. The sedentary occupation of office workers, in turn, favored diseases of affluence. This might be particularly relevant for long-term occupied persons and older workers due to a hormonal shift coming along, given the risk for oxidative stress-related diseases such as cardiovascular disease, particularly in the case of males, based on their lifestyle habits.

Oxidized LDL Increase the Proinflammatory Profile of Human Visceral Adipocytes Produced by Hypoxia.

BACKGROUND: Little is known about the effects of hypoxia on scavenger receptors (SRs) levels in adipocytes. We analyzed the effect of morbid obesity and hypoxia on SRs and inflammation markers in human visceral adipocytes and whether ox-LDL modify the inflammatory profile produced by hypoxia. METHODS: We studied in 17 non-obese and 20 subjects with morbid obesity (MO) the mRNA expression of HIF-1α, SRs (LOX-1, MSR1, CL-P1 and CXCL16), IL6 and TNFα in visceral adipocytes and the effect of hypoxia with or without ox-LDL on visceral in vitro-differentiated adipocytes (VDA). RESULTS: HIF-1α, TNFα, IL6, LOX-1, MSR1 and CXCL16 expression in adipocytes was increased in MO when compared with those in non-obese subjects (p < 0.05). The expression of most of the inflammatory markers and SRs gene correlated with HIF-1α. In VDA, hypoxia increased TNFα, IL6, MSR1, CXCL16 and CL-P1 (p < 0.05) in non-obese subjects, and TNFα, IL6, MSR1 and CXCL16 (p < 0.05) in MO. Silencing HIF-1α prevented the increase of TNFα, IL6, LOX-1, MSR1, CL-P1 and CXCL16 expression (p < 0.05). The combination of hypoxia and ox-LDL produced higher TNFα expression (p = 0.041). CONCLUSIONS: Morbid obesity and hypoxia increased SRs and inflammatory markers in visceral adipocytes. In a hypoxic state, ox-LDL increased the proinflammatory response of visceral adipocytes to hypoxia.

Preliminary Findings on the Association of the Lipid Peroxidation Product 4-Hydroxynonenal with the Lethal Outcome of Aggressive COVID-19.

Major findings of the pilot study involving 21 critically ill patients during the week after admission to the critical care unit specialized for COVID-19 are presented. Fourteen patients have recovered, while seven passed away. There were no differences between them in respect to clinical or laboratory parameters monitored. However, protein adducts of the lipid peroxidation product 4-hydroxynonenal (HNE) were higher in the plasma of the deceased patients, while total antioxidant capacity was below the detection limit for the majority of sera samples in both groups. Moreover, levels of the HNE-protein adducts were constant in the plasma of the deceased patients, while in survivors, they have shown prominent and dynamic variations, suggesting that survivors had active oxidative stress response mechanisms reacting to COVID-19 aggression, which were not efficient in patients who died. Immunohistochemistry revealed the abundant presence of HNE-protein adducts in the lungs of deceased patients indicating that HNE is associated with the lethal outcome. It seems that HNE was spreading from the blood vessels more than being a consequence of pneumonia. Due to the limitations of the relatively small number of patients involved in this study, further research on HNE and antioxidants is needed. This might allow a better understanding of COVID-19 and options for utilizing antioxidants by personalized, integrative biomedicine approach to prevent the onset of HNE-mediated vitious circle of lipid peroxidation in patients with aggressive inflammatory diseases.

Coating with Hypertonic Saline Improves Virus Protection of Filtering Facepiece Manyfold-Benefit of Salt Impregnation in Times of Pandemic.

Recently, as is evident with the COVID-19 pandemic, virus-containing aerosols can rapidly spread worldwide. As a consequence, filtering facepieces (FFP) are essential tools to protect against airborne viral particles. Incorrect donning and doffing of masks and a lack of hand-hygiene cause contagion by the wearers' own hands. This study aimed to prove that hypertonic saline effectively reduces the infectious viral load on treated masks. Therefore, a hypertonic salt solution´s protective effect on surgical masks was investigated, specifically analyzing the infectivity of aerosolized Alphacoronavirus 1 in pigs (Transmissible Gastroenteritis Virus (TGEV)). Uncoated and hypertonic salt pre-coated FFPs were sprayed with TGEV. After drying, a defined part of the mask was rinsed with the medium, and the eluent was used for the infection of a porcine testicular cell line. Additionally, airborne microorganisms´ long-term infectivity of sodium-chloride in phosphate-buffered saline comprising 5% saccharose was investigated. In the results from an initial Median Tissue Culture Infectious Dose, infection rate of TGEV was minimally reduced by untreated FFP. In contrast, this could be reduced by a factor of 104 if FFPs were treated with hypertonic salt solutions. Airborne pathogens did not contaminate the growth medium if salt concentrations exceeded 5%. We conclude that hypertonic saline is a vital tool for anti-virus protection, exponentially improving the impact of FFPs.

The Onset of Systemic Oxidative Stress Associated with the Accumulation of Lipid Peroxidation Product Acrolein in the Skin of Patients with Small-Vessel Vasculitis.

Small-vessel vasculitis (SVV) is the inflammation of the vessel wall that can result in hemorrhage and/or ischemia. Among the histological findings in SVV are increased infiltrating neutrophils, which, due to their oxidative burst and myeloperoxidase activity, release excessive reactive oxygen species, triggering a chain reaction of lipid peroxidation and yielding reactive aldehydes such as acrolein. The implication of oxidative stress in the pathogenesis of SVV was studied, focusing on acrolein immunohistochemistry in the affected skin vessels and systemic stress response. Samples from SVV patients and healthy subjects were collected and analyzed for total serum peroxides, total antioxidant capacity, inflammatory and immunological parameters, as well as for the presence of acrolein-protein adducts in the skin tissue specimens. The obtained data showed that systemic redox homeostasis and iron metabolism are altered in SVV patients. Possible biomarkers in the evaluation of oxidative status, disease activity and prevalence were indicated. Furthermore, a strong correlation between the accumulation of acrolein-protein adducts in the skin and the progression of the disease was revealed. Thus, the results of this study demonstrate that SVV is not only associated with systemic oxidative stress but also with tissue-specific oxidative stress that promotes acrolein formation and protein modification correlating with the severity of cutaneous vasculitis.

Improved protection of filtering facepiece through inactivation of pathogens by hypertonic salt solutions - A possible COVID-19 prevention device.

The filtering facepiece operates through filtration without the ability to kill the viruses. If the filtration might be combined with antiviral agents simultaneously in the masks, this would be much more efficient during the use of these masks and against cross-infection after being discarded. For centuries, sodium chloride (NaCl) contributes to inhibiting pathogens on various occasions. If aerosol with infectious agents reaches the filtering face-piecé surface of the filtering face-piece, coated with hypertonic saline, they become attracted by hygroscopic salt crystals. Proteins and nucleic acids lose their structural integrity and become inactivated concerning their infectious properties. We provide further evidence for cell growth inhibition with hypertonic saline in yeast cells comprising a defending cell wall. Proliferation was inhibited in a concentration-dependent manner, i.e., above 50 g/L, yeast cell proliferation was completely blocked. At a NaCl concentration of 100 g/L, even decomposition of the original inoculated organisms was observed. Therefore, we conclude that hypertonic saline- coated filtering facepiece might strongly reduce the numbers of infectious particles on their surfaces and thus protect mask carriers efficiently from infections.

Oxidized LDL Modify the Human Adipocyte Phenotype to an Insulin Resistant, Proinflamatory and Proapoptotic Profile.

Little information exists in humans on the regulation that oxidized low-density lipoprotein (oxLDL) exerts on adipocyte metabolism, which is associated with obesity and type 2 diabetes. The aim was to analyze the oxLDL effects on adipocytokine secretion and scavenger receptors (SRs) and cell death markers in human visceral adipocytes. Human differentiated adipocytes from visceral adipose tissue from non-obese and morbidly obese subjects were incubated with increasing oxLDL concentrations. mRNA expression of SRs, markers of apoptosis and autophagy, secretion of adipocytokines, and glucose uptake were analyzed. In non-obese and in morbidly obese subjects, oxLDL produced a decrease in insulin-induced glucose uptake, a significant dose-dependent increase in tumor necrosis factor-α (TNF-α), IL-6, and adiponectin secretion, and a decrease in leptin secretion. OxLDL produced a significant increase of Lox-1 and a decrease in Cxcl16 and Cl-p1 expression. The expression of Bnip3 (marker of apoptosis, necrosis and autophagy) was significantly increased and Bcl2 (antiapoptotic marker) was decreased. OxLDL could sensitize adipocytes to a lower insulin-induced glucose uptake, a more proinflammatory phenotype, and could modify the gene expression involved in apoptosis, autophagy, necrosis, and mitophagy. OxLDL can upregulate Lox-1, and this could lead to a possible amplification of proinflammatory and proapoptotic effects of oxLDL.

[Bioavailability of natural versus synthetic B vitamins and their effects on metabolic processes]. / Bioverfügbarkeit eines natürlichen versus eines synthetischen Vitamin-B- Komplexes und deren Auswirkungen auf metabolische Prozesse.

BACKGROUND: Owing to the widespread use of vitamin supplements to prevent and compensate for deficiencies, the equivalence of natural versus synthetic vitamins with respect to their bioavailability and metabolic influence is discussed controversially. METHOD: Thirty healthy female (n=22) and male participants (n=8) were investigated in a randomized, double-blind, cross-over study over a supplementation period of 6 weeks for each condition. The participants received a daily dose of a complex of the 8 natural B vitamins (group N), determined by the natural composition of quinoa seedlings, resp. synthetic B vitamins (group S), both corresponding to about 2.5 times the Recommended Dietary Allowance (RDA) of the national nutrition board. The primary criterion under investigation was changes in the blood levels of the individual B vitamins. Secondary criteria were the influence of both B complexes on homocysteine, antioxidant status, polyphenols, peroxide loading and peroxidase activity. RESULTS: Compared to baseline values, serum levels of all B vitamins measured increased: Vitamins B1 (N +23%; S +27%), B2 (N +14%; S +13%), B6 (N +101%; S +101%), B9 (N +86%; S +153%) and B12 (N +16%) were elevated at the end of the first supplementation period (p < 0.05), while serum levels of vitamins B1, B9 and B12 remained elevated compared to baseline even after the 2-week washout phase. During the second supplementation period, the vitamin concentrations in group N, with the exception of vitamin B1, could be increased once again (p < 0.05). In contrast, in group S only for vitamins B2 and B12 substantial increases (p < 0.05) were found. The influence of B vitamins on metabolic parameters such as homocysteine and polyphenols, which were markedly reduced, was also clearly measurable; however, total antioxidant capacity and peroxidase activity increased. The peroxide concentration remained almost unchanged in both groups. CONCLUSION: This clinical pilot study showed comparable bioavailability for both natural and synthetic B vitamins, with a 2.5-fold concentration of the RDA. Both vitamin B preparations showed a clear influence on metabolic parameters, whereas that of the natural B vitamins tended to have a slightly stronger effect than the synthetic analogues.

A Micromethod for Polyphenol High-Throughput Screening Saves 90 Percent Reagents and Sample Volume.

There is ample evidence that polyphenols are important natural substances with pronounced antioxidative properties. This study aimed to develop a fast and reliable method to determine total polyphenol content (TPC) in foodstuffs and human samples. The microtitration format offers the advantage of low sample volumes in the microlitre range, facilitating high-throughput screening with 40 samples simultaneously. We accordingly adjusted the so-called Folin-Ciocalteu method to a microtitre format (polyphenols microtitre-PPm) with 90% reduction of reagents. The assay was standardized with gallic acid in the range between 0.1 and 3 mM, using a 20 µL sample volume. The intra-assay coefficient of variation (CV) was less than 5%, and inter-assay CV was in the range of 10%. Wavelength was measured at 766 nm after two hours of incubation. This micromethod correlates significantly with both the classical Folin-Ciocalteu method and High-Performance Thin-Layer Chromatography (HPTLC) (r2 = 0.9829). We further observed a significant correlation between PPm and total antioxidants (r2 = 0.918). The highest polyphenol concentrations were obtained for red, blue, and black fruits, vegetables, and juices. Extracts of red grapes could be harvested almost sugar free and might serve as a basis for polyphenol supplementation. Beer, flour, and bread contained polyphenol concentrations sufficient to meet the minimal daily requirement. We conclude that PPm is a sensitive and reliable method that detects polyphenols even in samples diluted 10-fold. The literature strongly recommends further investigations on the effects of polyphenol uptake on human and animal health.

A Randomized Pilot Trial to Evaluate the Bioavailability of Natural versus Synthetic Vitamin B Complexes in Healthy Humans and Their Effects on Homocysteine, Oxidative Stress, and Antioxidant Levels.

The vitamin B complex comprises 8 different water-soluble constituents that humans must sequester from the diet. This pilot study compared natural versus synthetic vitamin B complexes for their bioavailability, accumulation, and their impact on antioxidants, homocysteine levels, and oxidative stress. We conducted a double-blind randomized clinical trial with thirty healthy participants. They were randomly assigned to group N (natural) and group S (synthetic). Vitamin B was ingested daily for 6 weeks in the range of about 2.5 times above the recommended daily allowance. Blood samples were taken at baseline, 1.5 h, 4 h, 7 h (diurnal), 6 w (discontinuation of supplements), and 8 w (washout). Blood levels of thiamine (B1), riboflavin (B2), pyridoxine (B6), folic acid (B9), cobalamin (B12), homocysteine, total antioxidants, peroxidase activity, polyphenols, and total peroxides were determined. Compared to initial values, serum levels of each B vitamin increased at the end of the supplementation period: i.e., B1 (+23% N; +27% S), B2 (+14% N; +13% S), B6 (+101% N; +101% S), B9 (+86% N; +153% S), and B12 (+16% N) (p < 0.05). Homocysteine (-13% N) decreased, while peroxidase activity (+41% S) and antioxidant capacity increased (+26% N). Short-term effects were already observed after 1.5 h for B9 (+238% N; +246% S) and after 4 h for vitamin B2 (+7% N; +8% S), B6 (+59% N; +51% S), and peroxidase activity (+58% N; +58% S). During the washout period, serum levels of B vitamins decreased except for thiamine and peroxidase activity, which increased further. This clinical pilot study revealed comparable bioavailability for both natural and synthetic B vitamins but did not show statistically noticeable differences between groups despite some favourable tendencies within the natural vitamin group, i.e., sustained effects for cobalamin and endogenous peroxidase activity and a decrease in homocysteine and oxidative stress levels.

Antioxidative 1,4-Dihydropyridine Derivatives Modulate Oxidative Stress and Growth of Human Osteoblast-Like Cells In Vitro.

Oxidative stress has been implicated in pathophysiology of different human stress- and age-associated disorders, including osteoporosis for which antioxidants could be considered as therapeutic remedies as was suggested recently. The 1,4-dihydropyridine (DHP) derivatives are known for their pleiotropic activity, with some also acting as antioxidants. To find compounds with potential antioxidative activity, a group of 27 structurally diverse DHPs, as well as one pyridine compound, were studied. A group of 11 DHPs with 10-fold higher antioxidative potential than of uric acid, were further tested in cell model of human osteoblast-like cells. Short-term combined effects of DHPs and 50 µM H2O2 (1-h each), revealed better antioxidative potential of DHPs if administered before a stressor. Indirect 24-h effect of DHPs was evaluated in cells further exposed to mild oxidative stress conditions induced either by H2O2 or tert-butyl hydroperoxide (both 50 µM). Cell growth (viability and proliferation), generation of ROS and intracellular glutathione concentration were evaluated. The promotion of cell growth was highly dependent on the concentrations of DHPs used, type of stressor applied and treatment set-up. Thiocarbatone III-1, E2-134-1 III-4, Carbatone II-1, AV-153 IV-1, and Diethone I could be considered as therapeutic agents for osteoporosis although further research is needed to elucidate their bioactivity mechanisms, in particular in respect to signaling pathways involving 4-hydroxynoneal and related second messengers of free radicals.

Work Intensity, Low-Grade Inflammation, and Oxidative Status: A Comparison between Office and Slaughterhouse Workers.

Limited knowledge exists about the impact of physical workload on oxidative stress in different occupational categories. Thus, we aimed to investigate the oxidative and inflammatory status in employees with different physical workloads. We enrolled a total of 79 male subjects, 27 office workers (mean age 38.8 ± 9.1 years) and 52 heavy workers, in a slaughterhouse (mean age 40.8 ± 8.2 years). Fasting blood was drawn from an antecubital vein in the morning of the midweek before an 8-hour or 12-hour work shift. The antioxidative capacity was assessed measuring total antioxidant capacity (TAC), uric acid, total polyphenols (PPm), and endogenous peroxidase activity (EPA). Total peroxides (TOC), malondialdehyde (MDA), and myeloperoxidase (MPO) were analyzed as prooxidative biomarkers, and an oxidative stress index (OSI) was calculated. In addition, hsCRP, interleukin-6 (IL-6), MDA-LDL IgM antibodies, galectin-3, adrenocorticotropic hormone (ACTH), and the brain-derived neurotrophic factor (BDNF) were measured as biomarkers of chronic systemic inflammation and emotional stress. TOC (p = 0.032), TAC (p < 0.001), ACTH (p < 0.001), OSI (p = 0.011), and hsCRP (p = 0.019) were significantly increased in the heavy workers group, while EPA, BDNF (p < 0.001), and polyphenols (p = 0.004) were significantly higher in office workers. Comparison between 8 and 12 h shifts showed a worse psychological condition in heavy workers with increased levels for hsCRP (p = 0.001) and reduced concentration of BDNF (p = 0.012) compared to office workers. Oxidative stress and inflammation are induced in heavy workers and are particularly pronounced during long working hours, that is, 12-hour versus 8-hour shifts.

Cultivation and Immortalization of Human B-Cells Producing a Human Monoclonal IgM Antibody Binding to MDA-LDL: Further Evidence for Formation of Atherogenic MDA-LDL Adducts in Humans In Vivo.

Oxidatively modified low-density lipoprotein (oLDL) is firmly believed to play an important role in the initiation and development of atherosclerosis, and malonic dialdehyde (MDA) is one of the major lipid peroxidation breakdown products involved in this process. In recent decades, antibodies against MDA-LDL have been detected in human and animal sera. In our study, human B-cells from the peripheral blood of a healthy female donor were fused with the SP2/0 mouse myeloma cell line. Antibody-producing hybridomas were detected by MDA-LDL-IgG/IgM enzyme-linked immunosorbent assays (ELISA) and Cu++-oxidized LDL IgG/IgM (oLAb) ELISA. Cells with supernatants emitting positive signals for antibodies were then cloned and after sufficient multiplication frozen and stored under liquid nitrogen. Due to the loss of antibody-producing ability, we established an MDA-LDL-IgM-producing cell line by recloning. This allowed isolation and immortalization of several human B-cells. The human donor had not been immunized with MDA-modified proteins, thus obviously producing MDA-LDL antibodies in vivo. Furthermore, using these antibodies for in vitro experiments, we were able to demonstrate that MDA epitopes are among the epitopes generated during Cu++-LDL oxidation as well. Finally, these antibodies compete in ELISA and cell culture experiments with MDA as a challenging toxin or ligand.

Trace elements and oxidative stress in hypertensive disorders of pregnancy.

PURPOSE: Due to increased metabolic requests, pregnancy can be considered as metabolic stress, especially if associated with oxidative stress triggered by disbalance of pro/antioxidants. The aim of the study was to determine serum concentrations of the trace elements iron (Fe), zinc (Zn) and copper (Cu) important in growth regulation and pro/anti-oxidant homeostasis, in relation to the total serum oxidant capacity (TOC) and total serum antioxidant capacity (TAC) in pregnant women with preeclampsia (n = 30) or with gestational hypertension (n = 30) and in healthy pregnant women (n = 37) and non-pregnant women (n = 30) as control groups expecting common differences between all pregnant women and controls and between preeclampsia and the other pregnancies indicating specific disbalance of the oxidative stress and analyzed trace elements. METHODS: Serum Fe was determined by spectrophotometric method, Cu and Zn were determined by atomic absorption spectrometry, TOC was determined by Enzymatic ANTIOX-CAP assay and TAC by Peroxide-activity assay. RESULTS: Serum Cu and TOC were significantly higher while Zn was lower in all pregnant groups regardless of hypertensive disorders. Serum Fe and TAC concentrations were found to be significantly higher in pregnant women with preeclampsia compared to pregnant controls. CONCLUSION: Increase of TOC in all pregnant women our study points to latent oxidative stress in pregnancy. Fe might have a role in etiopathogenesis of preeclampsia while the increase of TAC in the very beginning of preeclampsia might represent a stressdefence mechanism of the body. It has still to be revealed whether significantly higher serum Fe levels are associated with preeclampsia as a cause or as a consequence of this disorder.

Lipidome analysis in multiple sclerosis reveals protein lipoxidative damage as a potential pathogenic mechanism.

Metabolomic and lipidomic analyses have been used for the profiling of neurodegenerative processes, both in targeted and untargeted approaches. In this work we have applied these techniques to the study of CSF samples of multiple sclerosis (MS) patients (n = 9), compared with samples of non-MS individuals (n = 9) using mass-spectrometry. We have used western-blot and analyzed cell culture to confirm pathogenic pathways suggested by mass-spectrometric measurements. The results of the untargeted approach of metabolomics and lipidomics suggest the existence of several metabolites and lipids discriminating both populations. Applying targeted lipidomic analyses focused to a pathogenic pathway in MS, oxidative stress, reveal that the lipid peroxidation marker 8-iso-prostaglandin F2α is increased in CSF from MS patients. Furthermore, as lipid peroxidation exerts its pathogenical effects through protein modification, we studied the incidence of protein lipoxidation, revealing specific increases in carboxymethylated, neuroketal and malondialdehyde-mediated protein modifications in proteins of CSF from MS patients, despite the absence of their precursors glyoxal and methylglyoxal. Finally, we report that the level of neuroketal-modified proteins correlated with a hitherto unknown increased amount of autoantibodies against lipid peroxidation-modified proteins in CSF, without compensation by signaling induced by lipid peroxidation via peroxisome proliferator-activated receptor γ (PPARγ). The results, despite the limitation of being obtained in a small population, strongly suggest that autoimmunity against in situ produced epitopes derived from lipid peroxidation can be a relevant pathogenic factor in MS.

Effects of bioreactive acrolein from automotive exhaust gases on human cells in vitro.

Acrolein is a toxic unsaturated aldehyde and widespread environmental pollutant produced during lipid peroxidation and also by burning of tobacco or liquid fuels. Inhalation or dermal exposure to acrolein could be toxic to organisms. This very reactive aldehyde has a strong affinity for binding to proteins thus forming pathogenic protein-adducts. In the present study we have analyzed formation of bioreactive acrolein-protein adducts in bovine serum albumin solution exposed to exhaust gases of mineral diesel fuel and of mineral diesel fuel supplemented with different amounts of a novel diesel fuel additive denoted Ecodiesel (produced by a genuine procedure of recycling of plant oils used for food preparation). The effects of acrolein-protein adducts were tested on human microvascular endothelial cells and on human osteosarcoma cells that are sensitive to bioactivities of lipid peroxidation products. The results have shown a reduction of the bioreactive acrolein in exhaust gases when mineral diesel was supplemented with 5-20% Ecodiesel. Moreover, acrolein-protein adducts obtained from mineral diesel supplemented with Ecodiesel were less toxic than those obtained from mineral diesel alone. Thus, we assume that supplementing mineral diesel fuel with Ecodiesel would be of benefit for the use of renewable energy, for environment and for human health due to reduced environmental pollution with bioreactive acrolein.

Ergometry induces systemic oxidative stress in healthy human subjects.

Oxidative stress is an important pathogenic factor of cancer and cardiovascular, metabolic and degenerative diseases. On the other hand, mild oxidative stress, as in case of physical exercise, can increase the antioxidant defense system. However, the mechanisms underlying such desirable effects of mild oxidative stress are not well understood, because the production of hydroxyl radical, the most aggressive oxygen free radical, was not yet evaluated under physiological circumstances. Therefore, in this study, we evaluated the overall production of hydroxyl radical using blood samples of ten healthy male students before and 1 h after ergometry. One h before exercise, they took salicylic acid (1g) orally so that hydroxyl radical was trapped with salicylic acid, yielding a measurable reaction product, 2,3-dihydroxybenzoic acid. Oxidative stress response to exercise was also evaluated in the volunteers without premedication by measuring serum peroxides and total antioxidant capacity of serum. These parameters of oxidative stress were then correlated with physical performance of the subjects. Ergometry caused an increase of the plasma hydroxyl radical level by 37.5% (p < 0.05), whereas the levels of total serum peroxides did not change significantly. Total serum antioxidant capacity, measured as uric acid equivalents, was higher after ergometry by 39.7% (p < 0.05), and was in positive correlation (r = 0.81) with anaerobic threshold, an indicator of physical condition. Hence, ergometry induces hydroxyl radical production and systemic oxidative stress response in the healthy subjects. Egometry could be used to study physiological oxidative stress response and to improve antioxidant defense capacities in humans.