利用电子自旋共振（ESR）技术研究一氧化氮自由基在心脑血管疾病和健康中的“双刃剑”作用

一氧化氮(NO)是一个简单的气体自由基,在心血管、免疫和神经系统是一个重要的信号分子．NO由不同形式的NO合酶,内皮型NO合酶(eNOS),神经型NO合酶(nNOS)和诱导型NO合酶(iNOS)在不同细胞中产生．NO在调节血压和血管稳态方面发挥重要的生理作用,但是,由iNOS产生过量的NO就会抑制心脏收缩、损伤线粒体甚至引起细胞凋亡．NO可以影响神经递质释放及突出体生成过程和可塑性,NO在脑的发育和维持调节脑回路、作为学习的信号分子发挥着重要作用．然而,越来越多的证据表明,如果NO产生过多,就可能损伤心血管和神经,甚至引起心脑血管还推行型疾病．该文综述了近年来利用ESR研究NO在心脑血管疾病和健康方面的“双刃剑”作用和天然抗氧化剂的保护作用的一些结果．     

Loss of molars early in life develops behavioral lateralization and impairs hippocampus-dependent recognition memory

Background

Using senescence-accelerated mouse prone 8 (SAMP8), we examined whether reduced mastication from a young age affects hippocampal-dependent cognitive function. We anesthetized male SAMP8 mice at 8 weeks of age and extracted all maxillary molar teeth of half the animals. The other animals were treated similarly, except that molar teeth were not extracted. At 12 and 24 weeks of age, their general behavior and their ability to recognize novel objects were tested using the open-field test (OFT) and the object-recognition test (ORT), respectively.

Results

The body weight of molarless mice was reduced significantly compared to that of molar-intact mice after the extraction and did not recover to the weight of age-matched molar-intact mice throughout the experimental period. At 12 weeks of age, molarless mice showed significantly greater locomotor activity in the OFT than molar-intact mice. However, the ability of molarless mice to discriminate a novel object in the ORT was impaired compared to that of molar-intact mice. The ability of both molarless and molar-intact SAMP8 mice to recognize objects was impaired at 24 weeks of age. These results suggest that molarless SAMP8 mice develop a deficit of cognitive function earlier than molar-intact SAMP8 mice. Interestingly, both at 12 and 24 weeks of age, molarless mice showed a lateralized preference of object location in the encoding session of the ORT, in which two identical objects were presented. Their lateralized preference of object location was positively correlated with the rightward turning-direction preference, which reached statistical significance at 24 weeks of age.

Conclusions

Loss of masticatory function in early life causes malnutrition and chronic stress and impairs the ability to recognize novel objects. Hyperactivation and lateralized rotational behavior are commonly observed with dysfunction of the dopaminergic system, therefore, reduced masticatory function may deplete the mesolimbic and mesocorticolimbic dopaminergic systems to impair the cognitive functions of selective attention and recognition memory in the prefrontal cortex and the hippocampus.     

Sleep,aging, and lifespan in <Emphasis Type="Italic">Drosophila</Emphasis>

Background  

Epidemiological studies in humans suggest that a decrease in daily sleep duration is associated with reduced lifespan, but this issue remains controversial. Other studies in humans also show that both sleep quantity and sleep quality decrease with age. Drosophila melanogaster is a useful model to study aging and sleep, and inheriting mutations affecting the potassium current Shaker results in flies that sleep less and have a shorter lifespan. However, whether the link between short sleep and reduced longevity exists also in wild-type flies is unknown. Similarly, it is unknown whether such a link depends on sleep amount per se, rather than on other factors such as waking activity. Also, sleep quality has been shown to decrease in old flies, but it remains unclear whether aging-related sleep fragmentation is a generalized phenomenon.     

Early iNOS impairment and late eNOS enhancement during reperfusion following 2.49 MHz continuous ultrasound exposure after ischemia

ObjectiveIschemia reperfusion (IR) injury, occurring during heart attacks, hemorrhagic shock, and bypass and transplant surgeries, impairs microcirculatory function and nitric oxide (NO) synthesis. We report the regulation of endothelial and inducible NO synthase (eNOS and iNOS) proteins as a consequence of the application of continuous mode diagnostic frequency ultrasound application following IR injury.MethodsAnimals were assigned to one of five groups for microcirculatory assessment or Western blot analysis (WB) as follows: (1) IR+iNOS inhibition (1400W); and (2) IR+1400W+ultrasound for microcirculatory assessment, (3) Control; (4) IR; and (5) IR+ultrasound for WB. Functional capillary density and microvascular diameter, flow velocity, and flow were monitored for microcirculatory assessment. Skin tissue samples were harvested for WB. 2.49 MHz continuous ultrasound was used for application.ResultsBoth the inhibition of iNOS alone and iNOS inhibition with ultrasound irradiation positively influenced the microcirculation of observed animals relative to baseline values. Ultrasound exposure resulted in a significant production of eNOS protein in skin tissue harvested 24 h into reperfusion (p < 0.01). iNOS levels from the same tissue of ultrasound exposed animals were found to be significantly decreased 0.5 h into reperfusion (p < 0.05).ConclusionProtection from lasting IR injury effects in the microcirculation, with continuous mode diagnostic frequency ultrasound, results from augmented eNOS protein levels during late reperfusion. Ultrasound inhibited iNOS protein production during early reperfusion may also confer protection from IR injury.     

Estrous cycle influences the expression of neuronal nitric oxide synthase in the hypothalamus and limbic system of female mice

Background  

Nitric oxide plays an important role in the regulation of male and female sexual behavior in rodents, and the expression of the nitric oxide synthase (NOS) is influenced by testosterone in the male rat, and by estrogens in the female. We have here quantitatively investigated the distribution of nNOS immunoreactive (ir) neurons in the limbic hypothalamic region of intact female mice sacrificed during different phases of estrous cycle.     

Evidence for oxidative stress in the developing cerebellum of the rat after chronic mild carbon monoxide exposure (0.0025% in air)

Background  

The present study was designed to test the hypothesis that chronic very mild prenatal carbon monoxide (CO) exposure (25 parts per million) subverts the normal development of the rat cerebellar cortex. Studies at this chronic low CO exposure over the earliest periods of mammalian development have not been performed to date. Pregnant rats were exposed chronically to CO from gestational day E5 to E20. In the postnatal period, rat pups were grouped as follows: Group A: prenatal exposure to CO only; group B: prenatal exposure to CO then exposed to CO from postnatal day 5 (P5) to P20; group C: postnatal exposure only, from P5 to P20, and group D, controls (air without CO). At P20, immunocytochemical analyses of oxidative stress markers, and structural and functional proteins were assessed in the cerebellar cortex of the four groups. Quantitative real time PCR assays were performed for inducible (iNOS), neuronal (nNOS), and endothelial (eNOS) nitric oxide synthases.     

Immunological impact of magnetic nanoparticles (Ferucarbotran) on murine peritoneal macrophages

Ferucarbotran, a clinically used superparamagnetic iron oxide, is widely developed as a magnetic resonance imaging (MRI) contrast agent and has the potential to improve the monitoring of macrophage recirculation in vivo. However, the biological effect of Ferucarbotran or magnetic nanoparticles (MNPs) on macrophage is not clearly understood yet. This study is aimed to examine the immunological impact of Ferucarbotran toward murine peritoneal macrophages. Cells treated with Ferucarbotran demonstrated a dose–responsive increase of granularity in the cytoplasm. After 24 h of incubation, viability and cytotoxicity in macrophages treated with 200 μg Fe/mL of Ferucarbotran were not affected. Macrophages loaded with Ferucarbotran above 100 μg Fe/mL showed a significant (p < 0.01) increase in cytokine (TNF-α, IL-1β, IL-6) secretion and mRNA expression, followed by nitric oxide (NO) secretion and iNOS mRNA expression. Chemotactic responses of Ferucarbotran-preloaded macrophages toward CX3CL1 were significantly (p < 0.05) lower than those of untreated macrophages. Taking together, Ferucarbotran at high dose (100 μg Fe/mL) could induce murine peritoneal macrophages activation in pro-inflammatory cytokine secretion and NO production.     

EPR Study of the Intensity of the Nitric Oxide Production in Rat Brain After Ischemic Stroke

Electron paramagnetic resonance of (DETC)₂–Fe²⁺–NO complexes has been used as a method to detect the formation of nitric oxide (NO) in the brain tissues of Wistar rats. The content of nitric oxide in the center of ischemia (left brain hemisphere), in the non-ischemic part of the left hemisphere, and also in the regions conditionally not affected by ischemia, the cerebral cortex of the right hemisphere and the cerebellum of rats was studied in 5, 9, 24 and 72 h after modeling an ischemic stroke. It is found that in the ischemic part of the left hemisphere in 5 h after modeling the ischemic stroke, the NO content in the spin trap and R-conformer compositions decreases by 5 times and 30%, respectively. This decrease in the NO content is found in 9 and 24 h after the stroke. In the cerebral cortex of the right hemisphere and non-ischemic parts of the cerebral cortex of the left hemisphere, the NO content in the composition of the spin trap in 5, 9 and 24 h after the stroke decreases by 40–50%. In the composition of the R-conformer it does not vary. In 72 h, the partial restoration of the NO content in all regions except for the ischemia zone is observed.     

A role for cryptochromes in sleep regulation

Background  

The cryptochrome 1 and 2 genes (cry1 and cry2) are necessary for the generation of circadian rhythms, as mice lacking both of these genes (cry1,2 ^-/-) lack circadian rhythms. We studied sleep in cry1,2 ^-/- mice under baseline conditions as well as under conditions of constant darkness and enforced wakefulness to determine whether cryptochromes influence sleep regulatory processes.     

A non-circadian role for clock-genes in sleep homeostasis:a strain comparison

Background  

We have previously reported that the expression of circadian clock-genes increases in the cerebral cortex after sleep deprivation (SD) and that the sleep rebound following SD is attenuated in mice deficient for one or more clock-genes. We hypothesized that besides generating circadian rhythms, clock-genes also play a role in the homeostatic regulation of sleep. Here we follow the time course of the forebrain changes in the expression of the clock-genes period (per)-1, per2, and of the clock-controlled gene albumin D-binding protein (dbp) during a 6 h SD and subsequent recovery sleep in three inbred strains of mice for which the homeostatic sleep rebound following SD differs. We reasoned that if clock genes are functionally implicated in sleep homeostasis then the SD-induced changes in gene expression should vary according to the genotypic differences in the sleep rebound.     

Everyday episodic memory in amnestic mild cognitive impairment: a preliminary investigation

Background

In humans, rapid eye movements (REM) density during REM sleep plays a prominent role in psychiatric diseases. Especially in depression, an increased REM density is a vulnerability marker for depression. In clinical practice and research measurement of REM density is highly standardized. In basic animal research, almost no tools are available to obtain and systematically evaluate eye movement data, although, this would create increased comparability between human and animal sleep studies.

Methods

We obtained standardized electroencephalographic (EEG), electromyographic (EMG) and electrooculographic (EOG) signals from freely behaving mice. EOG electrodes were bilaterally and chronically implanted with placement of the electrodes directly between the musculus rectus superior and musculus rectus lateralis. After recovery, EEG, EMG and EOG signals were obtained for four days. Subsequent to the implantation process, we developed and validated an Eye Movement scoring in Mice Algorithm (EMMA) to detect REM as singularities of the EOG signal, based on wavelet methodology.

Results

The distribution of wakefulness, non-REM (NREM) sleep and rapid eye movement (REM) sleep was typical of nocturnal rodents with small amounts of wakefulness and large amounts of NREM sleep during the light period and reversed proportions during the dark period. REM sleep was distributed correspondingly. REM density was significantly higher during REM sleep than NREM sleep. REM bursts were detected more often at the end of the dark period than the beginning of the light period. During REM sleep REM density showed an ultradian course, and during NREM sleep REM density peaked at the beginning of the dark period. Concerning individual eye movements, REM duration was longer and amplitude was lower during REM sleep than NREM sleep. The majority of single REM and REM bursts were associated with micro-arousals during NREM sleep, but not during REM sleep.

Conclusions

Sleep-stage specific distributions of REM in mice correspond to human REM density during sleep. REM density, now also assessable in animal models through our approach, is increased in humans after acute stress, during PTSD and in depression. This relationship can now be exploited to match animal models more closely to clinical situations, especially in animal models of depression.     

Dynamic changes in GABA<Subscript>A</Subscript>receptors on basal forebrain cholinergic neurons following sleep deprivation and recovery

Background  

The basal forebrain (BF) cholinergic neurons play an important role in cortical activation and arousal and are active in association with cortical activation of waking and inactive in association with cortical slow wave activity of sleep. In view of findings that GABA_A receptors (Rs) and inhibitory transmission undergo dynamic changes as a function of prior activity, we investigated whether the GABA_ARs on cholinergic cells might undergo such changes as a function of their prior activity during waking vs. sleep.     

Ion aging effects on the dissociative-attachment instability in CO2 lasers

Onset of the dissociative-attachment instability requires that the rate coefficient for electron detachment (k _d) from negative ions be below a critical value. The predominant negative and positive ions in a CO₂N₂He gas-discharge plasma are known to change with time. As secondary by-products form and the predominant negative-ion species changes from CO ₃ ^– to NO ₂ ^– , a decrease ink _d occurs destabilizing the discharge. Since NO ₂ ^– and NO ₃ ^– are largely unreactive with respect to associative detachment,k _d depends in a sensitive fashion on the concentration of certain minority negative ions (O^–, O ₂ ^– ) and neutrals (CO, O, N). The sufficient conditions for the dissociative-attachment instability are much less sensitive to changes in the electron-ion and ion-ion recombination rate coefficients resulting from the ion aging process.     

NO dissociation and N2 production in the presence of co-adsorbed SO2 and D2 on Pt(3 3 2)

NO dissociation and subsequent N₂ production in the presence of co-adsorbed S¹⁸O₂ and D₂ on the surface of stepped Pt(3 3 2) were studied using Fourier transform infra red reflection–absorption spectroscopy (FTIR-RAS) combined with thermal desorption spectroscopy (TDS). Reduction of NO by D (D₂ is adsorbed dissociatively on Pt surfaces) proceeds to a limited extent, because this reaction is rate-controlled by NO dissociation and the supply of D atoms at the higher surface temperatures at which NO dissociation becomes significant (350 K and higher). NO–D reaction is suppressed in the presence of S¹⁸O₂, depending significantly on the S¹⁸O₂ coverage and the competition between the reactions NO–D and S¹⁸O₂–D. When the supply of D₂ is limited, e.g., 0.1 L in this study, the presence of S¹⁸O₂ suppresses the NO–D reaction. With a sufficient supply of D₂, e.g., 0.4 L and higher, D-atom competing reactions do not play a role any more because the reactions of both NO and S¹⁸O₂ with D proceed only to a very limited extent. As such, generation of O atoms from S¹⁸O₂ dissociation is the main reaction that leads to the suppression in NO dissociation and consequently, N₂ production.It is also concluded that the presence of S¹⁸O₂ does not seriously poison the active sites on the Pt surface, providing that there is a sufficient D supply to remove O atoms from both NO dissociation and S¹⁸O₂ dissociation.     

Ultrafast dissociation processes in the NO dimer studied with time-resolved photoelectron imaging

Ultraviolet photodissociation of the NO dimer is studied with femtosecond time-resolved photoelectron imaging (TR-PEI) spectroscopy. Pump pulses in the range 200–235 nm are employed, while probe pulses are kept at 300 nm. The time dependencies of the observed photoelectron kinetic energies and photoelectron angular distributions support a picture in which valence state optically excited in the dimer evolves on a time scale of <1 ps to the dimer 3s Rydberg state. This dimer Rydberg state then undergoes fragmentation on a time scale of a few ps. In this study we focus on dissociation into an NO ground state fragment and an NO fragment in its 3s Rydberg A²Σ⁺ state. Every stage of this continuous process, viz. the dimer valence state, the dimer 3s Rydberg state, the separating NO(X) + NO(A) fragments, and the isolated NO(A) fragment is interrogated with TR-PEI.     

Numerical study on effects of hydrothermal aging to composite regeneration in CeO2-based catalyzed diesel particulate filter

The effects of hydrothermal aging on the performance of CeO₂-based catalyzed diesel particulate filter (CDPF) was numerically investigated in this study based on a zero-dimensional model using the plug flow reactor in which a chemical reaction kinetic mechanism was established and validated by the simulated gas environment experiment. The effects of regeneration temperature, O₂ concentration in the ultimate emission conditions, the ratio of NO₂ in NOx (α) and the ratio of NOx to soot (β) on catalyst deactivation temperature and soot oxidation rate were investigated with fresh and hydrothermal aging CeO₂-based CDPF. The results show that hydrothermal aging of CeO₂-based catalysts raises the regeneration temperature from 613 to 783 K and shifts the soot catalytic combustion reaction path from complete to incomplete oxidation. Soot oxidation rate of fresh catalyst first increases rapidly at 516 K and then starts to slow down gradually at 633 K, but for hydrothermal aging catalysts, are 601 K and 789 K, respectively. With O₂ concentration increased from 1.5 mol/m³ to 5.5 mol/m³, the catalyst deactivation of fresh and hydrothermal aging catalyst increased from 609 K to 602 K, 791 K to 818 K, respectively. The increase in α and β leads to an increase in soot oxidation rate and a decrease in regeneration temperature. The deactivation temperature of catalyst is increased in higher α (1.0) and lower β (0.1), which the highest is 821 K. Synergistic mechanisms of NOx, regeneration temperature, and hydrothermal aging effects on soot catalytic combustion in CeO₂-based CDPF are revealed deeply with the help of zero-dimensional model.     

Non-rapid eye movement sleep with low muscle tone as a marker of rapid eye movement sleep regulation

Background  

It was recently reported that epochs of non-REM sleep (NREMS) with low muscle tone represent a partial correlate of REM sleep (REMS). To further investigate this phenomenon, episodes of restricted night-time sleep (23:00 – 03.00h) and subsequent morning sleep (10:00 – 13:00h) were analysed.     

Elevated intracellular chloride level in albino visual cortex neurons is mediated by Na-K-Cl co-transporter

Background  

During development the switch from a depolarizing to a hyperpolarizing action of GABA is a consequence of a decrease of the Na⁺-K⁺-2Cl^- co-transporter (NKCC1, Cl^--uptake) and increase of the K⁺-Cl^- co-transporter (KCC2, Cl^--extrusion) expression. However albino visual cortex neurons don't show a corresponding decrease in intracellular chloride concentration during development of the visual system as compared to pigmented animals.     

Infrared laser-spectroscopic analysis of <Superscript>14</Superscript>NO and <Superscript>15</Superscript>NO in human breath

We report on monitoring of nitric oxide (NO) traces in human breath via infrared cavity leak-out spectroscopy. Using a CO sideband laser near 5 μm wavelength and an optical cavity with two high-reflectivity mirrors (R=99.98%), the minimum detectable absorption is 2×10⁻¹⁰ cm⁻¹ Hz^1/2. This allows for spectroscopic analysis of rare NO isotopologues with unprecedented sensitivity. Application to simultaneous online detection of ¹⁴NO and ¹⁵NO in breath samples collected in the nasal cavity is described for the first time. We achieved a noise-equivalent detection limit of 7 parts per trillion for nasal ¹⁵NO (integration time: 70 s).