Reaction mechanisms of flavins and flavoproteins from an electronic-structure perspective / Mecanismos de reação de flavinas e flavoproteínas por uma perspectiva de estrutura eletrônica

Proteins equipped with flavin adenine dinucleotides (FAD) or flavin mononucleotides (FMN) are named flavoproteins and constitute about 1% of all existing proteins. They catalyze redox, acid-base and photochemical reactions in a variety of biochemical phenomena that goes from energy metabolism to DNA repair and light sensing. The versatility observed in flavoproteins is ultimately a balance of flavin intrinsic properties modulated by a protein environment. This thesis aims to investigate how flavoproteins work by systematic evaluating flavin properties and reactivity. In particular, the mechanism of fumarate reduction by the flavoenzyme fumarate reductase Fcc3 was determined. Electronic-structure calculations were used for this task based on rigorous calibration with experimental data and error assessment. Flavin properties at chemical accuracy were obtained with single reference coupled-cluster CCSD(T) calculations at the complete basis set limit. Density functional theory was demonstrated an excellent alternative with lower computational costs and slightly less accuracy. Flavin protonation and tautomerism were shown to be important modulators of flavin properties and reactivity, with the possibility of various tautomers existing at neutral pH. Regarding flavin redox properties, an analysis based on multiconfigurational wave function weights was proposed for categorizing flavin redox reactions as hydride or hydrogen-atom transfers. This analysis is an upgrade over traditional partial charges methods and can be applied not only to flavin reactions but to any protoncoupled electron transfer. In the investigation of the enzymatic mechanism of fumarate reduction, the reaction was determined as a nucleophilic addition by hydride transfer with carbanion formation. Fumarate reductase employs electrostatic catalysis in contrast to previous proposals of substrate straining and general-acid catalysis. Also, hydride transfer was shown to be vibronically adiabatic with low tunneling contribution. These findings give new insights into the mechanisms of fumarate reductases and provide a framework for future computational studies of flavoproteins in general. The analyses and benchmark studies presented can be used to build better models of properties and reactivity of flavins and flavoproteins

Proteínas equipadas com dinucleotídeos de flavina-adenina (FAD) e mononucleotídeos de flavina (FMN) são chamadas flavoproteínas e constituem cerca de 1% de todas as proteínas existentes. Elas catalisam reações redox, ácido-base e fotoquímicas numa variedade de fenômenos bioquímicos que vão desde o metabolismo energético até reparo de DNA e captação de luz. A versatilidade observada em flavoproteínas é em última instância um balanço das propriedades intrínsecas de flavinas moduladas por um ambiente proteico. Esta tese busca investigar como flavoproteínas funcionam através de avaliações sistemáticas de propriedades e reatividade de flavinas. Em particular, o mecanismo de redução de fumarato pela flavoenzima fumarato redutase Fcc3 foi determinado. Cálculos de estrutura eletrônica foram usados para esta tarefa com base em rigorosa calibração com dados experimentais e avaliação de erros. As propriedades de flavinas foram determinadas com acurácia química com cálculos monoconfiguracionais de coupled-cluster CCSD(T) no limite de conjunto base completo. A teoria do funcional da densidade mostrou-se uma alternativa excelente com menor custo computacional e um pouco menos de acurácia. Protonação e tautomerismo de flavinas mostraram-se moduladores importantes de suas propriedades e reatividade, com a possibilidade de vários tautômeros existirem em pH neutro. Em relação às propriedades redox de flavinas, uma análise baseada nos pesos de funções de onda multiconfiguracionais foi proposta para categorizar as reações redox de flavinas como transferências de hidreto ou hidrogênio. Esta análise é uma melhoria em relação aos métodos tradicionais de cargas parciais e pode ser aplicada não apenas para reações de flavinas mas para qualquer transferência de próton acoplada a elétrons. Na investigação do mecanismo enzimático de redução de fumarato, a reação foi designada como uma adição nucleofílica por transferência de hidreto e formação de carbânion. A fumarato redutase usa catálise eletrostática diferentemente de prospostas anteriores envolvendo distorção do substrato e catálise ácida geral. Além disso, a transferência de hidreto mostrou-se vibronicamente adiabática com pouca contribuição de tunelamento. Estas descobertas abrem novas perspectivas sobre os mecanismos de fumarato redutases e fornecem uma base para estudos computacionais futuros sobre flavoproteínas em geral. As análises e estudos comparativos apresentados podem ser usados para construir melhores modelos para propriedades e reatividade de flavinas e flavoproteínas

Can static electric fields increase the activity of nitric oxide synthase and induce oxidative stress and damage of spleen?

With the rapid development of ultra-high-voltage (UHV) direct-current (DC) transmissions, the impact of static electric fields (SEF) in the vicinity of overhead UHV DC transmission lines on health has aroused much public concern. This study explored the effects of 56.3kV/m SEF on the spleen of mice. Results showed that SEF exposure of 21days significantly increased malonic dialdehyde content, superoxide dismutase activity, calcineurin activity, nitric oxide synthase (NOS) activity, and the mRNA expression levels of tumor necrosis factor-α (TNF-α) and nuclear factor-κB (NF-κB) in the spleen and caused the separation of nucleus and nuclear membrane, the disappearance of mitochondrial membrane, and the deficiency of mitochondrial cristae in splenic lymphocytes. By analysis and discussion, it was deduced that SEF could induce oxidative stress of the spleen by increasing the activity of NOS. Oxidative stress could further cause ultrastructural changes of splenic lymphocytes. Moreover, oxidative stress could cause the increase of the mRNA expression levels of TNF-α and NF-κB, which contributed to the occurrence of spleen inflammation.

Water-Soluble Organic Nanoparticles with Programable Intermolecular Charge Transfer for NIR-II Photothermal Anti-Bacterial Therapy.

Extensive recent efforts have been put on the design of high-performance organic near-infrared (NIR) photothermal agents (PTAs), especially over NIR-II bio-window (1000-1350 nm). So far, the development is mainly limited by the rarity of molecules with good NIR-II response. Here, we report organic nanoparticles of intermolecular charge-transfer complexes (CTCs) with easily programmable optical absorption. By employing different common donor and acceptor molecules to form CTC nanoparticles (CT NPs), absorption peaks of CT NPs can be controllably tuned from the NIR-I to NIR-II region. Notably, CT NPs formed with perylene and TCNQ have a considerably red-shifted absorption peak at 1040 nm and achieves a good photothermal conversion efficiency of 42 % under 1064 nm excitation. These nanoparticles were used for antibacterial application with effective activity towards both Gram-negative and Gram-positive bacteria. This work opens a new avenue into the development of efficient PTAs.

A narrative review of hydrogen-oxygen mixture for medical purpose and the inhaler thereof.

Recent development regarding mixture of H2 (concentration of ~66%) with O2 (concentration of ~34%) for medical purpose, such as treatment of coronavirus disease-19 (COVID-19) patients, is introduced. Furthermore, the design principles of a hydrogen inhaler which generates mixture of hydrogen (~66%) with oxygen (~34%) for medical purpose are proposed. With the installation of the liquid blocking module and flame arresters, the air pathway of the hydrogen inhaler is divided by multiple isolation zones to prevent any unexpected explosion propagating from one zone to the other. An integrated filtering/cycling module is utilized to purify the impurity, and cool down the temperature of the electrolytic module to reduce the risk of the explosion. Moreover, a nebulizer is provided to selectively atomize the water into vapor which is then mixed with the filtered hydrogen-oxygen mix gas, such that the static electricity of a substance hardly occurs to reduce the risk of the explosion. Furthermore, hydrogen concentration detector is installed to reduce the risk of hydrogen leakage. Result shows that the hydrogen inhaler implementing the aforesaid design rules could effectively inhibit the explosion, even ignition at the outset of the hydrogen inhaler which outputs hydrogen-oxygen gas (approximately 66% hydrogen: 34% oxygen).

Electrostatic hazards of charging of bedclothes and ignition in medical facilities.

We investigated the charge generated on bedclothes (cotton and polyester) during bedding exchange with different humidities and the ignitability of an alcohol-based hand sanitizer (72.3 mass% ethanol) due to static spark with different temperatures to identify the hazards of electrostatic shocks and ignitions occurring previously in medical facilities. The results indicated that charging of the polyester bedclothes may induce a human body potential of over about 10 kV, resulting in shocks even at a relative humidity of 50%, and a human body potential of higher than about 8 kV can cause a risk for the ignition of the hand sanitizer. The grounding of human bodies via footwear and flooring, therefore, is essential to avoid such hazards (or to reduce such risks).

Studies on effects of static electric field exposure on liver in mice.

With the development of ultra-high-voltage direct-current transmission, the intensity of static electric field (SEF) under transmission lines increased, which has aroused public attention on its potential health effects. In order to examine effects of SEF exposure on liver, institute of cancer research mice were exposed to SEF with intensities of 27.5 kV/m, 34.7 kV/m and 56.3 kV/m, respectively. In each intensity of SEF exposure, a corresponding sham exposure group was used. Several indices relating to liver function (aspartate aminotransferase (AST) and alanine aminotransferase (ALT)) and oxidative stress (superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA)) were tested after exposure of 7, 14, 21 and 35 days. Results showed that exposure to SEF with intensities of 27.5 kV/m and 34.7 kV/m for 35 days did not significantly influence any detected indices above. Under SEF exposure with intensity of 56.3 kV/m, the SOD activity in liver was significantly increased after exposure of 7 and 14 days. However, no significant increase was found in MDA content as well as the activities of AST and ALT between exposure group and sham exposure group during SEF exposure of 56.3 kV/m. It suggested that from three SEF intensities, only exposure to SEF with intensity of 56.3 kV/m (7 and 14 days) caused a temporary oxidative stress response in liver expressed by the increase in activity of SOD, but it did not produce oxidative damage. This biological effect may be related to the increase of mitochondrial membrane potential of hepatocytes caused by SEF exposure. When the membrane potential exceeds a threshold, Q cycle in mitochondria will be affected, which will result in an increase of superoxide anion concentration and ultimately an oxidative stress.

Duration-dependent effect of exposure to static electric field on learning and memory ability in mice.

With the rapid development of ultra-high-voltage direct-current (UHVDC) transmission, the strength of environmental static electric field (SEF) around UHVDC transmission lines increased substantially, which has aroused widely public attention on the potential health effects of SEF. In this study, the effect of SEF exposure on learning and memory ability was investigated. Institute of Cancer Research mice were exposed to 56.3 kV/m SEF for a short term (7 days) or long term (49 days). Behaviors in the Morris water maze (MWM) test, hippocampal neurotransmitter contents, and oxidative stress indicators were examined. Results showed that short-term SEF exposure significantly prolonged escape latency and decreased the number of platform-site crossovers, as well as decreased the time spent in the target quadrant in the MWM test. Meanwhile, serotonin level and the ratio of glutamate level to γ-aminobutyric acid level changed significantly. Besides, malondialdehyde content and glutathione peroxidase activity increased significantly, while superoxide dismutase activity decreased significantly. After long-term SEF exposure, all indices above showed no significant differences between the SEF and sham exposure groups. These data indicated that short-term exposure to 56.3 kV/m SEF could cause abnormal neurotransmitter levels and oxidative stress in the hippocampus, which led to the decline in learning and memory ability. Under the condition of long-term exposure, the SEF-induced disturbances in neurotransmitter contents and redox balance were offset by the compensatory responses of mice, and thus, the learning and memory ability returned to normal level. The temporary and reversible decline in learning and memory ability was only a common biological effect of SEF rather than a health hazard.

Systematic review of biological effects of exposure to static electric fields. Part II: Invertebrates and plants.

BACKGROUND: The construction of high-voltage direct current (HVDC) lines for the long-distance transport of energy is becoming increasingly popular. This has raised public concern about potential environmental impacts of the static electric fields (EF) produced under and near HVDC power lines. As the second part of a comprehensive literature analysis, the aim of this systematic review was to assess the effects of static EF exposure on biological functions in invertebrates and plants and to provide the basis for an environmental impact assessment of such exposures. METHODS: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) was used to guide the methodological conduct and reporting. RESULTS: Thirty-three studies - 14 invertebrate and 19 plant studies - met the eligibility criteria and were included in this review. The reported behavioral responses of insects and planarians upon exposure strongly suggest that invertebrates are able to perceive the presence of a static EF. Many other studies reported effects on physiological functions that were expressed as, for example, altered metabolic activity or delayed reproductive and developmental stages in invertebrates. In plants, leaf damage, alterations in germination rates, growth and yield, or variations in the concentration of essential elements, for example, have been reported. However, these physiological responses and changes in plant morphology appear to be secondary to surface stimulation by the static EF or caused by concomitant parameters of the electrostatic environment. Furthermore, all of the included studies suffered from methodological flaws, which lowered credibility in the results. CONCLUSION: At field levels encountered from natural sources or HVDC lines (< 35kV/m), the available data provide reliable evidence that static EF can trigger behavioral responses in invertebrates, but they do not provide evidence for adverse effects of static EF on other biological functions in invertebrates and plants. At far higher field levels (> 35kV/m), adverse effects on physiology and morphology, presumably caused by corona-action, appear to be more likely. Higher quality studies are needed to unravel the role of air ions, ozone, nitric oxide and corona current on alterations in physiological functions and morphology.

Does static electric field from ultra-high voltage direct-current transmission lines affect male reproductive capacity? Evidence from a laboratory study on male mice.

With the development of ultra-high-voltage direct-current (UHVDC) transmission technology and increase in transmission voltage, the issue of environmental static electric field (SEF) pollution is standing out and its possible health effects have caused much public attention. In this study, the effects of chronic exposure to SEF on reproductive capacity of male mice were investigated. Twenty Institute of Cancer Research (ICR) mice were exposed to SEF (56.3 ± 1.4 kV/m, 49 days) generated by a high-voltage device. Several biological end points related to spermatogenesis and testicular function were evaluated, including reproductive organ coefficients, sperm motility and morphology, serum testosterone level, and testicular histology. No significant differences were found between the SEF-exposed and sham-exposed groups at the end of the exposure period. However, further observation through transmission electron microscopy revealed cristae losses in mitochondria of spermatogenic cells after SEF exposure. Nevertheless, the mitochondria injury did not affect sperm motility, which might be explained from the perspective of energy supply. That is, most of the energy required for sperm movement is generated by glycolysis which occurs in the cytoplasm rather than oxidative phosphorylation which occurs in mitochondria. In conclusion, this study indicates that exposure to SEF (56.3 ± 1.4 kV/m, 49 days) has limited effects on male reproductive capacity.

Lipoatrofia semicircular: 3 casos clínicos / Semicircular lipoatrophy: three clinical cases

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New Minimum Relative Humidity Requirements Are Expected to Lead to More Medical Device Failures.

In 2010, the Addendum D to ASHRAE Standard 170, "Ventilation of healthcare facilities," lowered the minimum relative humidity (RH) requirement of anesthetizing locations (including operating rooms, operating/surgical cystoscopic rooms, delivery rooms (Caesarean), recovery rooms, critical and intensive care, newborn intensive care, treatment rooms, trauma rooms (crisis or shock), laser eye rooms, newborn nursery suites, and endoscopy rooms) from 30 % to 20 %. The new minimum limit was adopted based on the results of a review paper that suggested that lowering humidity levels will have little or no impact on providing a safe environment for patients, staff, or medical equipment. That review paper reached this conclusion by assuming that there were no medical device failures due to electrostatic discharge (ESD). However, in an examination of the FDA's MAUDE database of reported defects and recalls, we identified numerous medical device failures explicitly due to ESD. This paper presents technical reliability and safety concerns regarding the new guidelines and recommends that such changes should not be implemented and that the guidelines should be revoked.

Semicircular lipoatrophy: an electrostatic hypothesis.

BACKGROUND: Semicircular lipoatrophy (SL) is a benign pathology characterized by subcutaneous adipose tissue atrophy. OBJECTIVE: To determine causes of SL. METHODS: We performed research on 42 cases of SL who had developed symptoms of lipoatrophy with a technical study of the building and clinical study of the affected patients. RESULTS: Electromagnetic field and electrostatic charge measurements were performed. No signals different from typical radio, TV and other wireless communication devices were recorded. Electrostatic charges were recorded in floors, environments and on different furniture; they were positive, and all charges disappeared on contact with the worker's skin. Data about the workplace were collected, and clinical examinations were performed, including blood and biochemistry tests and 18 ultrasound skin tests. CONCLUSIONS: As no abnormal electromagnetic measurements were found and electrostatic changes and occupational behaviors showed some relevant data and as after changes in order to control the electrostatic environment all of them improved, we can conclude that electrostatic charges but not electromagnetic ones are the main cause of SL.

An in vitro model of a system of electrical potential compensation in extracorporeal circulation.

OBJECTIVES: Extracorporeal circulation (ECC) in patients undergoing cardiac surgery induces systemic immune-inflammatory reaction that results in increased postoperative morbidity. Many factors are responsible for the adverse response after ECC. The present in vitro study aimed to investigate electric charges (ECs) generated during ECC, to set a device compensating the ECs, and checking its effect on red blood cells (RBC). MATERIALS AND METHODS: The electrical signals of blood in ECC were collected by a custom developed low-noise electronic circuit, processed by a digital oscilloscope (DSO) and a dynamic signal analyzer (DSA). The compensation of ECs was performed using a compensation device, injecting a nulling charge into the blood circuit. The compensation effect of the ECs on RBCs was evaluated by scanning electron microscope (SEM). RESULTS: The electrical analysis performed using both the DSO and the DSA confirmed the EC formation during ECC. The notable electric signals recorded in standard ECC circuits substantially nulled once the compensation device was used, thus confirming efficient EC compensation. After two hours of ECC, the SEM non-blended test on human RBC samples highlighted morphological changes in acanthocytes of the normal biconcave-shaped RBC. CONCLUSIONS: The outcomes confirm the development of parasitic ECs during ECC and that a suppressor system may decrease the potential damage of ECs. Nevertheless, further studies are ongoing in order to investigate the complex mechanisms related to lymphocytes and platelet morphological and physiological chances during triboelectric charges in ECC.

Control of surface charges by radicals as a principle of antistatic polymers protecting electronic circuitry.

Even minute quantities of electric charge accumulating on polymer surfaces can cause shocks, explosions, and multibillion-dollar losses to electronic circuitry. This paper demonstrates that to remove static electricity, it is not at all necessary to "target" the charges themselves. Instead, the way to discharge a polymer is to remove radicals from its surface. These radicals colocalize with and stabilize the charges; when they are scavenged, the surfaces discharge rapidly. This radical-charge interplay allows for controlling static electricity by doping common polymers with small amounts of radical-scavenging molecules, including the familiar vitamin E. The effectiveness of this approach is demonstrated by rendering common polymers dust-mitigating and also by using them as coatings that prevent the failure of electronic circuitry.

Lipoatrofia semicircular, una visión general a una patología laboral emergente en Chile / Semicircular lipatrophy, an overview of emerging labor pathology in Chile

La Lipoatrofia Semicircular (LS) es una enfermedad idiopática, que se caracteriza por una atrofia reversible y localizada, ya sea total o parcial, del tejido adiposo subcutáneo y que se ubica en zonas de contacto de la piel de las extremidades con el inmobiliario. Se caracteriza por presentarse en brotes endémicos, cuyo único factor en común es el lugar de trabajo. Sus causas son desconocidas, pero en la actualidad se proponen factores asociados como: el contacto de la zona afectada con inmobiliario, presencia de radiación electromagnética, presencia de descargas electroestáticas y humedad ambiental relativa baja. Un modelo actual para explicar la patogenia de esta enfermedad involucra la participación de descargas electrostáticas, que probablemente activarían la liberación de factor de necrosis tumoral a (TNF-a) y desencadenaría la fagocitosis de adipocitos. Actualmente no tiene un tratamiento médico, siendo las medidas preventivas de mitigación de los factores previamente mencionados las que producen una reversión de la lesión. Por otro lado, la presencia de estos brotes generan un gran impacto mediático por lo que también debe ser abordada comunicacionalmente. La presente revisión pretende sintetizar la literatura sobre el tema para exponer el conocimiento actual y lograr una noción de esta nueva enfermedad, tras su primera presentación en brote en nuestro país.

Lipoatrophia semicircularis (LS) (also known as "Semicircular lipoa-trophy") is an idiopathic disease, which is characterized by a reversible and localized atrophy, either total or partial, of subcutaneous adipose tissue and located in areas of skin contact with the real estate tips. It is characterized by appearing in endemic outbreaks whose only common factor is the workplace. Its causes are unknown, but currently associated factors have been proposed, such as: the contact area with the office furniture, the presence of electromagnetic radiation, electrostatic discharge and presence of low relative humidity. A current model to explain the pathogenesis of this disease involves the participation of electrostatic discharges, which probably activate the release of tumor necrosis factor a (TNF-a) and it would trigger phagocytosis of adipocytes. Currently there is no medical treatment, being proactive mitigation measures of the before mentioned factors, those that produce a reversion of the injury. On the other hand, the presence of these buds generates great media impact so it must be also addressed from a communication standpoint. The present review aims to summarize the literature on the subject to present current knowledge and achieve a notion of this new disease, after its first appearance in outbreak in our country.

Development of an electrostatic model predicting copper toxicity to plants.

The focus of the present study was to investigate the mechanisms for the alleviation of Cu toxicity in plants by coexistent cations (e.g. Al(3+), Mn(2+), Ca(2+), Mg(2+), H(+), Na(+), and K(+)) and the development of an electrostatic model to predict 50% effect activities (EA50s) accurately. The alleviation of Cu(2+) toxicity was evaluated in several plants in terms of (i) the electrical potential at the outer surface of the plasma membrane (PM) (Ψ(0)(°)) and (ii) competition between cations for sites at the PM involved in the uptake or toxicity of Cu(2+), the latter of which is invoked by the Biotic Ligand Model (BLM) as the sole explanation for the alleviation of toxicity. The addition of coexistent cations into the bulk-phase medium reduces the negativity of Ψ(0)(°) and hence decreases the activity of Cu(2+) at the PM surface. Our analyses suggest that the alleviation of toxicity results primarily from electrostatic effects (i.e. changes in both the Cu(2+) activity at the PM surface and the electrical driving force across the PM), and that BLM-type competitive effects may be of lesser importance in plants. Although this does not exclude the possibility of competition, the data highlight the importance of electrostatic effects. An electrostatic model was developed to predict Cu(2+) toxicity thresholds (EA50s), and the quality of its predictive capacity suggests its potential utility in risk assessment of copper in natural waters and soils.

Blood pressure modulation following activation of mast cells by cationic cell penetrating peptides.

Short cell penetrating peptides (CPP) are widely used in vitro to transduce agents into cells. But their systemic effect has not been yet studied in detail. We studied the systemic effect of the cell penetrating peptides, penetratin, transportan and pro-rich, on rat hemodynamic functions. Intra-arterial monitoring of blood pressure showed that injection of the positively charged penetratin and transportan in a wide range of concentrations (2.5-320 µg/kg) caused highly significant transient decrease in the systolic and diastolic blood pressure in a dose dependent manner (p<0.01). Pretreatment with histamine receptors blockers or with cromolyn, a mast cell stabilizing agent, significantly attenuated this effect. Furthermore, in vitro incubation of these both peptides with mast cells line, LAD2, caused a massive mast cell degranulation. In vitro studies showed that these CPP in a wide range of concentrations were not cytotoxic without any effect on the survival of LAD2 mast cell line. In contrast, the less positively charged and proline-rich CPP, pro-rich, had no systemic effects with no effect on mast cell degranulation. Our results indicate that intravenously administrated positively charged CPP may have deleterious consequences due to their induced BP drop, mediated by mast cell activation. Therefore, the major effect of mast cell activation on BP should be considered in developing possible future drug therapies based on the injection of membrane-permeable and positively charged CPP. Nevertheless, lower levels of such CPP may be considered as a treatment of systemic high BP through moderate systemic mast cell activation.