AKT/GSK-3ß signaling is altered through downregulation of mTOR during cerebral Ischemia/Reperfusion injury.

PURPOSE: Cellular responses following cerebral ischemia/reperfusion injury are critical to recovery and survival after ischemic stroke. Understanding of these cellular responses can help the design of therapies to protect brain tissue and promote recovery after stroke. One of these cellular responses may be mediated by the AKT (protein kinase B) signal transduction pathway. This study was aimed to investigate the cerebral ischemia-induced alterations of AKT signaling and the upstream molecular pathways. METHODS: We modeled cerebral ischemia by middle cerebral artery occlusion in 2-3-month-old male C57BL/6J mice and then analyze the brain samples by using quantitative Western blots and phosphorylation/activation-dependent kinase antibodies. Cerebral ischemia was confirmed by staining of brain slices with 1% 2,3,5-triphenyltetrazolium chloride (TTC) and Nissl, as well as neurological assessments of the mice 24 h after ischemia-reperfusion surgery. RESULTS: We found marked downregulation of AKT within 12 h of cerebral ischemia/reperfusion, which leads to overactivation of glycogen synthase kinase-3ß (GSK-3ß). Furthermore, we found that the downregulation of AKT was mediated by downregulation of mTORC2 (the complex 2 of the mechanistic target of rapamycin) instead of its common upstream kinases, phosphatidylinositol 3-kinase and phosphoinositide-dependent kinase-1. CONCLUSION: Our findings provide new insight into the cellular responses to ischemia/reperfusion brain injury and will help develop new treatments targeting the AKT signaling pathway for the treatment of ischemic stroke.

Sevoflurane-induced neuronal apoptosis in neonatal mice is prevented with intranasal administration of insulin.

Concerns about the potential neurotoxicity of general anesthesia to the developing brain have been increasing in recent years. Animal studies have shown that neonatal exposure to general anesthesia causes both acute neurotoxicity and behavioral abnormalities later in life. In the present study, we observed over-activation of neuronal apoptosis in the brain of neonatal mice after a single exposure to anesthesia with sevoflurane for 6 hours at the age of 7 days. More importantly, we found that insulin administered through intranasal delivery prior to anesthesia prevented anesthesia-induced over-activation of neuronal apoptosis. This study provides experimental evidence for a potential effective, yet simple, method to prevent anesthesia-induced neurotoxicity in children, especially in infants.

Intranasal Administration of Insulin Reduces Chronic Behavioral Abnormality and Neuronal Apoptosis Induced by General Anesthesia in Neonatal Mice.

Children, after multiple exposures to general anesthesia, appear to be at an increased risk of developing learning disabilities. Almost all general anesthetics-including sevoflurane, which is commonly used for children-are potentially neurotoxic to the developing brain. Anesthesia exposure during development might also be associated with behavioral deficiencies later in life. To date, there is no treatment to prevent anesthesia-induced neurotoxicity and behavioral changes. In this study, we anesthetized 7-day-old neonatal mice with sevoflurane for 3 h per day for three consecutive days and found that the anesthesia led to mild behavioral abnormalities later in life that were detectable by using the novel object recognition test, Morris water maze, and fear conditioning test. Biochemical and immunohistochemical studies indicate that anesthesia induced a decrease in brain levels of postsynaptic density 95 (PSD95), a postsynaptic marker, and marked activation of neuronal apoptosis in neonatal mice. Importantly, insulin administered through intranasal delivery prior to anesthesia was found to prevent the anesthesia-induced long-term behavioral abnormalities, reduction of PSD95, and activation of neuronal apoptosis. These findings suggest that intranasal insulin administration could be an effective approach to prevent the increased risk of neurotoxicity and chronic damage caused by anesthesia in the developing brain.

Remediation of petroleum contaminated soils by joint action of Pharbitis nil L. and its microbial community.

The plot-culture experiments were conducted for examining the feasibility of Pharbitis nil L. and its microbial community to remedy petroleum contaminated soils. The petroleum contaminated soil, containing 10% (w/w) of the total petroleum hydrocarbons (TPHs), was collected from the Shengli Oil Field, Dongying City, Shandong Province, China. The collected soil was applied and diluted to a series of petroleum contaminated soils (0.5%, 1.0%, 2.0% and 4.0%). Root length, microbial populations and numbers in the rhizosphere were also measured in this work. The results showed that there was significantly (p<0.05) greater degradation rate of TPHs in vegetated treatments, up to 27.63-67.42%, compared with the unvegetated controls (only 10.20-35.61%), after a 127-day incubation. Although various fractions of TPHs had an insignificant concentration difference due to the presence of the remediation plants, there was a much higher removal of saturated hydrocarbon compared with other components. The biomass of P. nil L. did not decrease significantly when the concentration of petroleum hydrocarbons in soil was ≤2.0%. The trends of microbial populations and numbers in the rhizosphere were similar to the biomass changes, with the exception that fungi at 0.5% petroleum contaminated soil had the largest microbial populations and numbers.

Promoted biodegradation and microbiological effects of petroleum hydrocarbons by Impatiens balsamina L. with strong endurance.

Phytoremediation is a promising green technology for cleanup of petroleum hydrocarbons (PHCs) in contaminated environment. Based on the objective of identifying special ornamental plants for the effective biodegradation of PHCs, the efficacy of Impatiens balsamina L. to phytoremedy petroleum contaminated soil from the Shengli Oil Field in Dongying City, Shandong Province, China, was further examined in a field plot-culture experiment under greenhouse conditions. After a 4-month culture period, the average degradation rate of total petroleum hydrocarbons (TPHs) by the plant was up to 18.13-65.03%, greatly higher than that (only 10.20-35.61%) in their corresponding controls by natural degradation. Among petroleum compositions saturated hydrocarbons had the highest degradation. The release of polar metabolic byproducts during phytoremediation of contaminated soils with ≥20,000 mg/kg of PHCs by I. balsamina may occur. Some growth indexes of I. balsamina indicated that the plant had a good tolerance to contaminated soils with ≤10,000 mg/kg of PHCs. Moreover rhizosphere bacteria and fungi became the dominant microbial population in soils with 5000 and 10,000 mg/kg of PHCs and were probably responsible for TPH degradation. Thus, I. balsamina L. could be a potential ornamental plant for effective phytoremediation of contaminated soils with ≤10,000 mg/kg of PHCs.

[Separation performance of 1-microm silica particles as stationary phase for pressurized electrochromatography].

Bare nonporous silica particles with diameter around 1 microm were prepared. A 20 cm section of a total length of 45 cm capillary (100 microm i. d.) was packed electrokinetically and the separation performance of basic compounds was investigated in the pressurized capillary electrochromatography (pCEC) system using 1 microm nonporous bare silica spheres as stationary phase and acetonitrile-water as mobile phase. The effects of the composition of the mobile phase, the concentration of the buffer, pH value, applied voltage and so on on the separation performance were investigated. The experimental results demonstrated that the bare silica column showed a typical reversed phase separation mechanism when it was used for separation of basic compounds. The separation performance changed slightly with changing buffer concentration. Since the degree of dissociation of the basic compounds depended greatly on the pH value, the interaction between the compounds and stationary phase changed with the changing of pH value, thereby changing the resolution of the compounds. The separation performance increased with the increase of applied voltage. A column efficiency of 35 000 for o-toluidine was obtained when a voltage of 1 kV was applied.

Silica spheres coated with C18-modified gold nanoparticles for capillary LC and pressurized CEC separations.

Nonporous monodispersed silica spheres of 1.3 microm were coated with gold nanoparticles (AuNPs) and subsequently coated with n-octadecanethiol. By transmission electron microscopy analysis, the average diameter of the AuNPs on the silica spheres was determined to be 12 nm. The chromatographic and electrochromatographic properties of self-assembled n-octadecanethiol AuNP-coated silica microspheres (C18-AuNPs-SiO2) were investigated using a group of nonpolar PAHs. The stationary phase appears to display a characteristic reversed-phase behavior. Higher separation efficiency and shorter separation times were obtained using pressurized CEC (pCEC) compared with capillary LC (CLC). A maximum column efficiency of about 2.5x10(5) plates per meter and less than 18 min separation time for benzene were obtained in pCEC while only 66 507 plates per meter and an analysis time of nearly 100 min were observed in CLC mode. A chemical stability test of the C18-AuNPs-SiO2 stationary phase under extremely high and low pH conditions demonstrated that it is stable at pH 12 and 1 for at least 60 h. The results confirm that C18-AuNPs-SiO2 possesses a high rigidity to withstand high packing pressures and can be used as a good stationary phase for CLC and pCEC.

Phytoremediation of petroleum contaminated soils by Mirabilis Jalapa L. in a greenhouse plot experiment.

Phytoremediation of soils contaminated by organic chemicals is a challenging problem in environmental science and engineering. On the basis of identifying remediation plants from ornamentals, the remediation capability of Mirabilis Jalapa L. to treat petroleum contaminated soil from the Shengli Oil Field in Dongying City, Shandong Province, China was further investigated using a field plot experiment carried out in a greenhouse. The results showed that the average efficiency of removing total petroleum hydrocarbons (TPHs) by M. jalapa over the 127-day culture period was high, up to 41.61-63.20%, when the removal rate by natural attenuation was only 19.75-37.92%. The maximum reduction occurred in the saturated hydrocarbon fraction compared with other components of petroleum contaminants. According to the qualitative and quantitative parameters including plant height, fresh weight, dry weight, root length, root weight and visual stress symptoms, it was indicated that M. jalapa had a peculiar tolerance to petroleum contamination and could effectively promote the degradation of TPHs when the concentration of petroleum hydrocarbons in soil was equal to and lower than 10,000 mg/kg. The population of living microorganisms in the planted soil could be also adaptive to

Toxic effects of crude-oil-contaminated soil in aquatic environment on Carassius auratus and their hepatic antioxidant defense system.

Under the indoor simulant conditions, toxic effects of crude-oil-contaminated soil which was put into aquatic environment on the young fishes Carassius auratus and their hepatic antioxidant system after a 20-d exposure were investigated. Results showed that the relationship between the mortality of C. auratus and the exposed doses could be divided into 3 phases: fishes exposed to the low dose groups (0.5-5.0 g/L) were dead due to the ingestion of crude-oil-contaminated soils in aquatic environment; at the medium dose groups (5.0-25.0 g/L) fishes were dead due to the penetration of toxic substances; at the high dose groups (25.0-50.0 g/L) fishes were dead due to environmental stress. The highest mortality and death speed were found in the 1.0 g/L dose group, and the death speed was sharply increased in the 50.0 g/L dose group in the late phase of exposure. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) and the content of malaondialdehyde (MDA) in the hepatic tissues of C. auratus were induced significantly. The activity of SOD was increased and then decreased. It was significantly inhibited in the 50.0 g/L dose group. The activity of CAT was highly induced, and restored to a level which is little more than the control when the exposed doses exceeded 10.0 g/L. The activity of GST was the most sensitive, it was significantly induced in all dose groups, and the highest elevation was up to 6 times in the 0.5 g/L dose group comparing with the control. The MDA content was significantly elevated in the 50.0 g/L dose group, and the changes of the MDA content were opposite with the changes of GST activity.