Calpain activity in adult and aged human brain regions

We assayed calpain activity in 27 human brain regions from adult (43–65 years of age) and aged (66–83 years of age) postmortem tissue samples. Calpain I (M Ca-requiring) activity was 10% or less of the total activity; it was below detectable levels in a number of areas, and so data are are expressed as total (M+mM Ca-dependent) calpain activity. The distribution of the enzyme was regionally heterogeneous. Highest activity was found in the spinal cord, followed by the amygdala, and levels in mesencephalic areas and in cerebellar grey matter were also high. Levels in cerebellar white matter, tegmentum, pons, and putamen were low, and activity in cortical areas was also relatively low. Although in some areas activity seemed higher with aging, the differences were not statistically significant. We previously found that the regional distribution of cathepsin D in human and in rat brain is similar, this seems to be true for calpain activity as well. The increase of protease activity with age found in rat brain is not found in human areas, as was shown previously with cathepsin D, and in the present study with calpain.Special issue dedicated to Dr. Bernard W. Agranoff.     

Combined low-frequency brain oscillatory activity and behavior predict future errors in human motor skill

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A new experimental animal for the study of age-related changes in serotonergic neuronal activity in the brain cortex

Calcium uptake by the cortical synaptosomes in a rodent (Fischer rat) and an insectivore shrew (Suncus murinus) was detected as a parameter reflecting molecular dysfunction of the aging brain. The change in calcium uptake by the cortical synaptosomes in both species was concomitant which showed less than half the capacity at 24 months old animals compared with those at 8 months old. On the other hand, 5-hydroxytryptamine binding and imipramine binding to the membrane fraction of aging rat brain cortex was not altered in terms of binding capacity along with aging, while, in Suncus, the binding of both serotonergic ligands declined with aging. In order to elucidate decreased serotonergic activity in human demented aged brain, together with declining activity in neurotransmitting systems detectable as a function of calcium uptake by the cortical synaptosomes, Suncus may be an appropriate animal model for studying physiological aging processes in the mammalian brain cortex.Special Issue Dedicated to Dr. Abel Lajtha.     

Nitric oxide synthase activity and the level of nitrates/nitrites in brain regions during spontaneous morphine withdrawal in rats

Activity of nitric oxide synthase (NOS) and concentrations of nitrate/nitrites (NO _x ^? ) were measured in brain regions of rats during spontaneous morphine withdrawal, which was modeled in male Wistar rats. The animals were injected with the increasing intraperitoneal doses (10–100 mg/kg, twice a day) of morphine hydrochloride for 6 days. Thirty six hours after the last injection the severity of the spontaneous morphine withdrawal syndrome was determined by specific autonomic and locomotor indices The withdrawal was accompanied by the increase of both NOS activity and NO _x ^? levels in the midbrain and hippocampus, the decrease of these parameters in striatum and hypothalamus, and lack of changes in cerebral cortex and brain stem. In cerebellum NOS activity decreased whereas NO _x ^? concentrations remained unchanged. In the cerebral cortex, striatum, midbrain, and cerebellum activity of NOS and NO _x ^? concentrations correlated with the withdrawal syndrome severity and also with the specific signs of abstinence.     

Alteration of neural activity and neuroinflammatory factors in the insular cortex of mice with corneal neuropathic pain

Dry eye disease (DED) affects nearly 55% of people worldwide; several studies have proposed that central sensitization and neuroinflammation may contribute to the developing corneal neuropathic pain of DED, while the underlying mechanisms of this contribution remain to be investigated. Excision of extra orbital lacrimal glands established the dry eye model. Corneal hypersensitivity was examined through chemical and mechanical stimulation, and open field test measured the anxiety levels. Restingstate fMRI is a method of functional magnetic resonance imaging (rs-fMRI) was performed for anatomical involvement of the brain regions. The amplitude of low-frequency fluctuation (ALFF) determined brain activity. Immunofluorescence testing and Quantitative real-time polymerase chain reaction were also performed to further validate the findings. Compared with the Sham group, ALFF signals in the supplemental somatosensory area, secondary auditory cortex, agranular insular cortex, temporal association areas, and ectorhinal cortex brain areas were increased in the dry eye group. This change of ALFF in the insular cortex was linked with the increment in corneal hypersensitivity (p < 0.01), c-Fos (p < 0.001), brain-derived neurotrophic factor (p < 0.01), TNF-α, IL-6, and IL-1β (p < 0.05). In contrast, IL-10 levels (p < 0.05) decreased in the dry eye group. DED-induced corneal hypersensitivity and upregulation of inflammatory cytokines could be blocked by insular cortex injection of Tyrosine Kinase receptor B agonist cyclotraxin-B (p < 0.01) without affecting anxiety levels. Our study reveals that the functional activity of the brain associated with corneal neuropathic pain and neuroinflammation in the insular cortex might contribute to dry eye-related corneal neuropathic pain.     

Roles of glutamate and GABA receptors in setting the developmental timing of spontaneous synchronized activity in the developing mouse cortex

Spontaneous, synchronized electrical activity (SSA) plays important roles in nervous system development, but it is not clear what causes it to start and stop at the appropriate times. In previous work, we showed that when SSA in neonatal mouse cortex is blocked by TTX in cultured slices during its normal time of occurrence (E17-P3), it fails to stop at P3 as it does in control cultured slices, but instead persists through at least P10. This indicates that SSA is self-extinguishing. Here we use whole-cell recordings and [Ca2+]i imaging to compare control and TTX-treated slices to isolate the factors that normally extinguish SSA on schedule. In TTX-treated slices, SSA bursts average 4 s in duration, and have two components. The first, lasting about 1 s, is mediated by AMPA receptors; the second, which extends the burst to 4 s and is responsible for most of the action potential generation during the burst, is mediated by NMDA receptors. In later stage (P5-P9) control slices, after SSA has declined to about 4% of its peak frequency, bursts lack this long NMDA component. Blocking this NMDA component in P5-P9 TTX-treated slices reduces SSA frequency, but not to the low values found in control slices, implying that additional factors help extinguish SSA. GABA(A) inhibitors restore SSA in control slices, indicating that the emergence of GABA(A)-mediated inhibition is another major factor that helps terminate SSA.     

A method for detecting the effect of magnetic field on activity changes of neuronal populations of Morimus funereus (Coleoptera, Cerambycidae)

Modification of a new method for detecting changes in the activities of neuronal population and the nearest neuron is described. Preliminary measurements of the influence of a static magnetic field (2 mT) on neuronal population activity on eight individuals of an endangered insect species Morimus funereus are included. Five minutes exposure produced both excitatory (5/8) and inhibitory (3/8) effect on the activity of neuronal population of M. funereus antennal lobe. However, when the reversibility of induced effects was quantitatively analyzed, our results showed that they were prevailingly irreversible: (7/8) for the population, (6/8) for the nearest neuron.     

Inhibition of thymidine kinase activity by hydroxyurea during highly and less proliferative regions of rat brain

Hydroxyurea, when injected intraperitoneally, exerted marked inhibition on the activity of thymidine kinase in 5 day old postnatal cerebellum and 15 day old embryonic cerebrum. However, it failed to show any sustained inhibition on thymidine kinase activity in 5 day old postnatal cerebrum. In this case, the marginal decrease of thymidine kinase activity noticed during early intervals reversed back to more than normal value at a later time interval. These results along with our earlier findings are taken to indicate the differential action of this drug on thymidine kinase activity in rapidly and slowly proliferating regions of rat brain     

Postnatal changes of cathepsin D activity in rat liver and brain

Total and specific activity of cathepsin D (EC. 3.4.23.5) were measured in rat liver and brain from 1 to 98 days of age. The activity of cathepsin D in the liver of adult and newborn rats was the same while in the rat brain it was higher in adult than in newborn rats. In the liver maximum specific activity of cathepsin D occurred on the 10th postnatal day and minimum on the fourth day of age. In the brain maximum specific activity of the enzyme occurred on the 14th postnatal day. Total activity of cathepsin D increased after birth in rat liver and brain. These results are discussed in relation to the functional role of cathepsin D in the rat liver and the brain.     

Dual metabotropic glutamate receptor signaling enables coordination of astrocyte and neuron activity in developing sensory domains

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Metabolite of tryptophan promoting changes in EEG signal and the oxidative status of the brain

Tryptophan is an essential amino acid precursor of neurotransmitter serotonin and triptamine. During its metabolism, indole‐3‐acetic acid (IAA) is generated; this substance presents both antioxidant and prooxidant effects in different biological systems in addition to hipoglicemic effects. To date, electroencephalography (EEG) has been used to evaluate the temporal effect of several substances in neurotransmission. The goal of this study was to characterize the effect of IAA in the brain by analysing the EEG signal and evaluate the oxidative status by means of biochemical parameters. The EEG was acquired by using a noninvasive method, and the brain electric signal was analysed by advanced digital signal processing techniques to determinate the energy signal filtered in different band frequencies. Furthermore, the oxidative status of the brain was investigated by measuring the activity of antioxidant enzymes and lipid peroxidation as well as blood glucose rates of the animals treated with different doses of IAA. Our results showed the relationship of IAA administration with changes in EEG signals. The oxidative status of the brain was modified by IAA after 14 days of treatment. Copyright © 2014 John Wiley & Sons, Ltd.     

北方旱作区沟垄二元覆盖技术研究进展

北方旱作区降水稀少,水分不足是限制作物生长的主要因素,沟垄二元覆盖技术可进一步提高降水的利用效率和作物生产水平,在国内外已备受关注.为深刻理解中国北方旱作区沟垄二元覆盖技术模式及研究进展,本文就沟垄二元覆盖技术的概念、研究现状、配套农机具、技术模式及其土壤生态与作物效应进行了综述;在总结现有研究的基础上,归纳了典型旱作区沟垄二元覆盖技术模式的类型、作业操作及应用效果;科学评价不同沟垄二元覆盖技术下土壤与作物的生态效应;指出了目前沟垄二元覆盖技术研究中所存在的问题,并提出相应的建议.综合国内外沟垄集雨栽培技术的研究进展,提出沟垄二元覆盖模式及技术体系今后的重点研究方向:1)建立适合北方不同旱作类型区及不同作物的垄沟比;2)重视土壤水分与温度、肥力等其他因子耦合性研究;3)探索最佳环保型覆盖材料;4)加强沟垄二元覆盖模式及体系的技术评价、技术推广与配套机具设计等方面的研究.     

Effect of 6-Aminonicotinamide on the activity of hexokinase and lactate dehydrogenase isoenzymes in regions of the rat brain

Changes in the activity of hexokinase and lactate dehydrogenase isoenzymes in the three brain regions and heart were studied in the 6-Aminonicotinamide-treated rats. Drug administration decreased the particulate hexokinase and lactate dehydrogenase activity, but increased the soluble hexokinase     

A method for monitoring of oxygen saturation changes in brain tissue using diffuse reflectance spectroscopy

Continuous measurement of local brain oxygen saturation (SO₂) can be used to monitor the status of brain trauma patients in the neurocritical care unit. Currently, micro‐oxygen‐electrodes are considered as the “gold standard” in measuring cerebral oxygen pressure (pO₂), which is closely related to SO₂ through the oxygen dissociation curve (ODC) of hemoglobin, but with the drawback of slow in response time. The present study suggests estimation of SO₂ in brain tissue using diffuse reflectance spectroscopy (DRS) for finding an analytical relation between measured spectra and the SO₂ for different blood concentrations. The P₃ diffusion approximation is used to generate a set of spectra simulating brain tissue for various levels of blood concentrations in order to estimate SO₂. The algorithm is evaluated on optical phantoms mimicking white brain matter (blood volume of 0.5–2%) where pO₂ and temperature is controlled and on clinical data collected during brain surgery. The suggested method is capable of estimating the blood fraction and oxygen saturation changes from the spectroscopic signal and the hemoglobin absorption profile.

     

Suboptimal temperature-dependent changes in the brain development and activity in Galleria mellonella larvae

The development of Galleria mellonella larvae is strongly affected by suboptimal temperature (18°C). One-day-old last-instar larvae react to 18°C with the arrest of further development for several months described as facultative larval diapause. The aim of this study was to find what type of changes, if any, in the brain correlate with the larval diapause induced by suboptimal temperature. Morphological analysis demonstrated the gradual inhibition of brain development. Paraldehyde-fuchsin (PAF) staining revealed cyclicity in the activity of the medial neurosecretory cells (M-NSC) in the larval brain. SDS-PAGE was used to examine the brain proteins of larvae reared at 30°C and at 18°C. The rate of protein synthesis in the brain of the last instar larvae kept at 18°C, measured as l -[³⁵S]methionine incorporation during 2-h incubation in vitro, was only about 40% of the value characteristic for this tissue during normal development (at 30°C). Despite decrease in the rate of total protein synthesis, suboptimal temperature induced an increase in the level of two major brain proteins: 112 and 84 kDa. In SDS-PAGE analysis, these two proteins appear 21–28 days after transfer to the lower temperature. Whether these proteins are specific for induction of larval diapause of Galleria mellonella remains to be further investigated. Arch. Insect Biochem. Physiol. 38:66–73, 1998. © 1998 Wiley-Liss, Inc.     

Purification and characterization of G proteins from human brain: modification of GTPase activity upon phosphorylation

Summary Three G proteins from human brain membranes were purified to near homogeneity by conventional techniques including preparative electrophoresis. These G proteins were characterized by their ability to bind GTP, GDP and GTP analogs. Two of these proteins have molecular weights of 50,000 (G₅₀) and 36,000 (G₃₆), as determined on SDS-gels. G₃₆ was ADP-ribosylated by pertussis toxin. Thus, G₅₀ could represent a G_sα subunit, whereas G₃₆ could be G_iα or G_oα. G₅₀ was phosphorylated by cAMP dependent protein kinase and protein kinase C. G₃₆ was phosphorylated by a protein kinase independent of calcium and phospholipid, a proteolytic product of protein kinase C, analogous to protein kinase M. Phosphorylation of G₃₆ by this protein kinase induced a dramatic decrease in its GTPase activity. The third G protein, of molecular weight 22,000 probably belongs to the group of monomeric G proteins possessing functional similarities withras gene products. The regulation of G proteins involving calcium-dependent and independent pathways is delineated.     

Beneficial effect of daidzin in dry eye rat model through the suppression of inflammation and oxidative stress in the cornea

Present investigation evaluates the effect of daidzin in dry eye rat model through the suppression of inflammation and oxidative stress in the cornea. Briefly, electron spine resonance was used for the estimation of radical scavenging activity of daidzin and COX Fluorescent Activity Assay Kit was used for the estimation of PGS activity. Dry eye rat model was developed by removing the lacrimal gland and effect of daidzin was evaluated in dry eye rat model by estimating the fluorescein score, tear volume and expressions of heme oxigenase (HO-1), TNF α, Interlukin 6 (IL-6), matrix metallopeptidase 9 (MMP-9) and PGS-2. Result of the present study suggested that daidzin possess tyrosyl radical scavenging activity and thereby decreases the oxidative stress. Activity of PGS significantly increases in dry eye which was inhibited by daidzin treatment due to competitive inhibition of PGS. It also recovers the tear volume in dry eye rat model in which lacrimal gland was removed. Thus corneal erosion was improved by daidzin in dry eye rat model. Thus present study concludes that treatment with daidzin protects the cornea in dry eye rat model by suppression inflammation and oxidative stress.     

The effects of adrenalectomy and thermal stress on glutamic acid decarboxylase activity in different regions of the rat brain

Glutamic acid decarboxylase (GAD) enzyme activity was measured in synaptosomes prepared from the hypothalamus, the hippocampus, the striatum and the cerebral cortex of control, adrenalectomized and rat exposed to a thermal stress. Adrenalectomy caused a statistically significant decrease in the enzyme activity in the striatum, while it had no effect in the other three brain areas. On the other hand, exposure to the thermal stress resulted in a dramatic increase of GAD specific activity in all brain areas examined. This thermal stress-induced increase in enzyme activity was observed in both non-operated and adrenalectomized animals, which implies that it is not mediated by glucocorticoids.Abbreviations used GAD glutamic acid decarboxylase - GABA -aminobutyric acid - AET 2-aminoethylisourethonium bromide - ADX adrenalectomized - rpm revolutions per minute     

The relationship between flash evoked potentials and evoked amplitude modulation patterns of an applied UHF electromagnetic field in the rat

Pilot studies demonstrate evidence that the electrophysiological processes associated with flash stimulation of the central nervous system (CNS) of rats, as seen in the recordings of visual-evoked potentials, may also be detectable using an ultrahigh frequency electromagnetic field (UHFEMF). Patterns of amplitude modulation of an applied UHFEMF, when recorded and averaged, show strong correlations with simultaneously recorded evoked potentials. The data support the hypothesis that the UHFEMF amplitude is altered in a dynamic fashion by the tissue's electrophysiological processes that are involved with the generation of CNS electric fields.     

Flight-correlated activity changes in neurons of the lateral accessory lobes in the brain of the locust Schistocerca gregaria

The study investigates activity changes in neurons of the lateral accessory lobes in the brain of the locust Schistocerca gregaria during wind-elicited tethered flight. Neurons with ascending projections from the ventral nerve cord to the lateral accessory lobes showed flight-associated excitations which were modulated in the flight motor rhythm. Descending neurons with ramifications in the lateral accessory lobes were tonically excited corresponding to flight duration. The onset of wind-elicited responses in the descending neurons preceded the onset of flight motor activity by 22–60 milliseconds. Neurons connecting the lateral accessory lobes with the central body, the anterior optic tubercles, or other brain areas showed a variety of responses including activity changes during flight initiation and flight termination. Activity in many of these neurons was less tightly coupled to the flight situation and often returned to background levels before flight was terminated. Most of the recorded neurons responded, in addition, to stationary visual stimuli. The results suggest that the lateral accessory lobes in the locust brain are integrative links between the central body, visual pathways, and the ventral nerve cord. The possible involvement of these brain areas in flight control is discussed.