Plasma biochemical responses of the omnivorous crucian carp (Carassius auratus) to crude cyanobacterial extracts

Healthy crucian carp (Carassius auratus) were treated by intraperitoneal (i.p.) injection of crude cyanobacterial extracts at two doses, 50 and 200 mu g MC-LR equiv kg(-1) BW. High mortality (100%) was observed within 60 h post injection in the high-dose group. In the treated fish, activities of four plasma enzymes, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH), all showed substantial increases, with both dose and time-dependent effects. These increases of enzyme activity indicate severe impairment occurred in the liver of crucian carp over time. Plasma concentrations of energy-related biomolecules including glucose (GLU), cholesterol (CHO), triglyceride (TG), and total protein (TP) showed marked changes in the high-dose group, possibly a nutritional imbalance correlated with the liver injury caused by intraperitoneal exposure to crude cyanobacterial extracts.

Hematological and plasma biochemical responses of crucian carp (Carassius auratus) to intraperitoneal injection of extracted microcystins with the possible mechanisms of anemia

Alterations in hematological indices such as decreases in blood cell counts (RBC), hematocrit (Ht) and hemoglobin (Hb) concentrations are key symptoms of anemia. However, few experiments were conducted to examine changes in hematological indices of fish exposed to microcystins that are believed to be fatal to circulatory systems of vertebrates. An acute toxicological experiment was designed to study hematological changes of crucian carp injected intraperitoneally (i.p.) with extracted microcystins at two doses, 50 and 200 mu g MC-LReqkg(-1) body weight. After being i.p. injected with microcystins, the fish exhibited behavioral abnormity. There were significant decreases in RBC in the high-dose group, and in Ht and Hb concentrations in both dose groups, while erythrocte sedimentation rate (ESR) significantly increased, indicating the appearance of normocytic anemia. There were no prominent changes in the three red cell indices, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH,), and mean corpuscular hemoglobin concentration (MCHC). Increases in blood urea nitrogen (BUN) and creatinine (CR) in both dose groups suggest the occurrence of kidney impairment. Alteration in blood indices was reversible at the low dose group. Conclusively, anemia induced by kidney impairment was a key factor to cause abnormity of swimming behaviors and high mortality of crucian carp. (c) 2007 Elsevier Ltd. All rights reserved.

Reproductive effects of prenatal exposure to nonylphenol on zebrafish (Danio rerio)

Mature female and male zebrafish were separated and exposed to nonylphenol (NP) at 0.1, 1, 10, 50, 100 and 500 mu g/L, respectively, for 3 weeks. Gonadosomatic index (GSI) in both sexes and vitellogenin (VTG) induction in males was measured as the bioindicators for the impairment to the parents. The results indicated that 50 mu g/L of NP was the non-observed effect concentration (NOEC) for GSI and VTG induction. Afterwards, the 50 mu g/L NP exposed females and males, and the control females and males were cross-wise pair-bred in the control water for one week to examine the reproductive effects. The embryonic cathepsin D (CAT D) activity, eggshell thickness, fecundity, hatching rate and malformation (vertebral column flexure) rate of offspring were determined in the four pair-bred groups. While endpoints remained unchanged in the groups with exposed males, prenatal exposure of females to 50 mu g/L of NP resulted in the impairment of reproduction in groups with exposed females including inhibition of CAT D activity (P < 0.05), decrease of eggshell thickness (by 23.6%) and elevation of malformation rate (P < 0.001). These results suggested NP could induce reproductive damage to zebrafish at NOEC for parents. The results also imply that alterations of CAT D activity and eggshell thickness may be more sensitive biomarkers to indicate the reproductive effects caused by endocrine disrupting chemicals. (c) 2005 Elsevier Inc. All rights is reserved.

Highly soluble and stable recombinant holo-phycocyanin alpha subunit expressed in Escherichia coli

C-phycocyanin (Cpc) is one of the phycobiliproteins with highly fluorescent and various pharmacological activities Holo-Cpc-alpha Subunit (holo-CpcA) expressed in Escherichia colt resulted in low yield and tended to aggregate after purification in this Study, we constructed a new plasmid coding holo-CpcA fused with hexahistidine and maltose-binding protein tag, which designated as HMCpcA. to Improve Its Solubility and stability without the Impairment of its spectra anti fluorescent properties HMCpcA was significantly more stable over time and a wider range of pH as compared to holo-CpcA. In addition. both the solubility and yields of HMCpcA increase significantly We here provided an example to demonstrate that MBP could also Improve the stability of the protein it fused while it has been reported as a soluble fusion partner before. This novel fluorescent protein will facilitate the large-scale production and be potentially applicable for the development Of fluorescent probes, as well as antioxidant agents (C) 2009 Elsevier B V. All rights reserved

维持性血液透析患者认知功能特点研究

Objective: To study the episodic memory, semantic memory, cognitive planning ability and inhibition ability in MHD patients. Method: Neuropsychological research methods such as Action memory of verb-object phrase, Trail Making Test (A and B), Verbal Fluency Test, Go-No/Go test and Stroop Color Naming Task were used to investigate Episodic Memory 、Semantic Memory、Executive Function of 40 MHD and 40 NC. Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale(SDS), Social Support Scale, Life Satisfaction Scale, and biochemical examination were applied and their relationships with cognitive function were analized. The mean age and education level of MHD group and NC group have no significant difference. Result: 1.Action memory of verb-object phrase differed significantly between MHD group and NC group. 2.Two tests of Verbal Fluency differed significantly between MHD group and NC group. 3.Trail Making Test A, Trail Making Test B, the baseline condition of Go-No/Go Test and Stroop Color Naming Test differed significantly between MHD group and NC group. 4.There is no significant difference between MHD group and NC group on the correct rate of No/Go Test and the baseline condition. Both groups showed Stroop Effect in Go-No/Go test, but MHD group performed significantly worse. 5.In Stroop Color Naming Task Test, NC group showed Stroop Effect, significant Repeated Distraction Promotion Effect and significant Negative Priming Effect,while MHD group showed only Stroop Effect and no Repeated Distraction Promotion Effect and no Negative Priming Effect. There is significant difference in Stroop Effect between MHD group and NC group. Conclusion: 1.Comparing with NC group, episodic memory, semantic memory, cognitive planning ability, and inhibition ability of MHD group were impaired significantly. 2.The pathological aging of Executive Function in MHD group showed: executive Function should be a unitary system. 3.Cognitive impairment is negatively correlated with serum creatinine, blood pressure and anxiety score in MHD patients; and is related with hemoglobin, hematocrit, social support and life satisfaction. Keyword: maintenance hemodialysis, episodic memory, semantic memory, cognitive planning, inhibition ability.

2型糖尿病患者认知功能特点及相关影响因素研究

Objective: Type 2 diabetes patients’ performances of action memory , semantic memory and working memory and the related factors were explored. Methods: 60 Type 2 diabetes patients were compared with 60 age and gender and level of education matched non-diabetes controls. Mood were tested by SAS and SDS, MMSE was used to test the basic cognitive function, Trail Making Test A and B, Verbal fluency test, Go-No/Go test, and Stroop color-word test were used to investigate the executive function of Type 2 diabetes patients and normal controls (NC). Patients’ GLU, TG, TCH, HbA1c, insulin and Cp were tested and correlated with their action memory and working memory. Results: There was no difference between NC group and Type 2 diabetes patients in MMSE scores. There is depression and anxiety mood in Type 2 diabetes patients. Type 2 diabetes patients get lower score in action memory test. Comparing to NC group, Type 2 diabetes patients performed significantly worse in Trail Making Test A and B and verbal fluency test. In Stroop Test, NC group showed significant Stroop Effect and Repeated Distraction Promotion Effect and Negative Priming Effect. However, In Type 2 diabetes group, only the Stroop Effect appeared, but no Repeated Distraction Promotion Effect and Negative Priming Effect. There is no difference between Type 2 diabetes and NC in Stroop Effect. In Go-No/Go test, both of two groups showed significant Stroop Effect, however, there was no difference between them. And also there is no difference on error rate of all levels between them. The course of disease, GL, HbA1c, TG, TCH, INS and Cp affected action memory and working memory. Conclusion: Type 2 diabetes patients’ action memory, semantic memory and working memory were partially impaired. Controlling the levels of GLU, TG and TCH can delay these kinds of impairment.

鸡胚吗啡用药对学习记忆和成瘾易感性的影响及其受体机制

Prenatal morphine exposure affects neural development of fetus by impairing learning and memory, and increasing susceptibility to morphine abuse. Because nervous systems have different developmental characteristics during different developmental stages, administration of morphine at different stages also has different effects on learning, memory, and susceptibility to morphine. Due to the precise developmental processes of neurotransmitter systems in chick embryo’s brain, and unique superiority of chick embryo model, the purpose of the present studies was to explore critical periods correlated to the memory impairment and the increasing susceptibility to morphine, via one-trial passive avoidance and conditioned place preference as behavior models. Then the possible roles of mu and delta opioid receptors as the possible mechanism were analyzed. Experiment 1 showed that injecting low dose of morphine (1 mg/kg) during the period embryonic 5 to 8 significantly impaired the function of learning and memory, worse than any other periods of the same treatment. Experiment 2 showed that injecting low dose of morphine during the period embryonic 17 to 20 significantly increased the susceptibility to morphine in the new-born chicks. The affected chicks acquired the morphine conditioned place preference more quickly, and maintained it much longer. Experiment 3 showed that during E5-8, injecting delta receptor antagonist naltrindole reversed the learning and memory impairment caused by morphine while delta receptor agonist DPDPE impaired learning and partial memory function. On the other hand, mu opioid receptors had little effect. As for E17-20, given naloxonazine can reverse the increases of susceptibility to morphine, and the mu receptor agonist DAGO cause the increases of susceptibility to morphine. Delta receptors have no effect. The above results demonstrated that prenatal morphine expousure at different developmental periods of chick embryo caused different influences on memory and susceptibility to morphine. That is, E5-8 is the critical period correlate to memory impairment; and E17-20 is the critical period correlate to susceptibility to morphine. Delta receptors were critical in learning and memory impairment while mu receptors in susceptibility.

汉语深层失读及词汇－语义系统的认知神经心理学研究

Chinese deep dyslexia is an important type of reading disorder that attracted many research interests. In the current thesis, three studies concerning Chinese deep dyslexia were performed: (1) clinical dominating and incidental symptoms were collected and their relationships were analyzed, and error scores of different character and word types compared; to verify typical reading model of alphabetic readers, and to develope a novel model for reading Chinese scripts; (2) based on these results, further neuropsychological analysis on the basic rules of lexical-semantic system and semantic distance were employed; (3) rehabilitation scheme were shaped to verify our research results. With cognitive neuropsychological methods, this study was mainly focused on deep dyslexic patients with brain impairment. The results were compared with those of normal people on rapid reading. Computer emulation was also used to describe reading process of patients. Both group analysis and case study were carried out. This study for the first time systematically investigated the clinical symptoms of Chinese deep dyslexia. A novel model was developed with a hypothesis that the sublexical pathway is composed of two parallel pathways: the phonetic sublexical pathway and the semantic sublexical pathway. Two characteristics of Chinese deep dyslexia were found compared with alphabetic deep dyslexia: (1) having no distinct word class effect and imagination effect; (2) the organization of Chinese lexical-semantic system has correlation with construct regulation, imagination and splitability of characters. Evoking of semantic correlation is stronger than phonetic correlation.

左中部梭状回枕颞沟内皮质对汉字加工的必要性、选择性和作用性质

A number of functional neuroimaging studies with skilled readers consistently showed activation to visual words in the left mid-fusiform cortex in occipitotemporal sulcus (LMFC-OTS). Neuropsychological studies also showed that lesions at left ventral occipitotemporal areas result in impairment in visual word processing. Based on these empirical observations and some theoretical speculations, a few researchers postulated that the LMFC-OTS is responsible for instant parallel and holistic extraction of the abstract representation of letter strings, and labeled this piece of cortex as “visual word form area” (VWFA). Nonetheless, functional neuroimaging studies alone is basically a correlative rather than causal approach, and lesions in the previous studies were typically not constrained within LMFC-OTS but also involving other brain regions beyond this area. Given these limitations, it remains unanswered for three fundamental questions: is LMFC-OTS necessary for visual word processing? is this functionally selective for visual word processing while unnecessary for processing of non-visual word stimuli? what are its function properties in visual word processing? This thesis aimed to address these questions through a series of neuropsychological, anatomical and functional MRI experiments in four patients with different degrees of impairments in the left fusiform gyrus. Necessity: Detailed analysis of anatomical brain images revealed that the four patients had differential foci of brain infarction. Specifically, the LMFC-OTS was damaged in one patient, while it remained intact in the other three. Neuropsychological experiments showed that the patient with lesions in the LMFC-OTS had severe impairments in reading aloud and recognizing Chinese characters, i.e., pure alexia. The patient with intact LMFC-OTS but information from the left visual field (LVF) was blocked due to lesions in the splenium of corpus callosum, showed impairment in Chinese characters recognition when the stimuli were presented in the LVF but not in the RVF, i.e. left hemialexia. In contrast, the other two patients with intact LMFC-OTS had normal function in processing Chinese characters. The fMRI experiments demonstrated that there was no significant activation to Chinese characters in the LMFC-OTS of the pure alexic patient and of the patient with left hemialexia when the stimuli were presented in the LVF. On the other hand, this patient, when Chinese characters were presented in right visual field, and the other two with intact LMFC-OTS had activation in the LMFC-OTS. These results together point to the necessity of the LMFC-OTS for Chinese character processing. Selectivity: We tested selectivity of the LMFC-OTS for visual word processing through systematically examining the patients’ ability for processing visual vs. auditory words, and word vs. non-word visual stimuli, such as faces, objects and colors. Results showed that the pure alexic patients could normally process auditory words (expression, understanding and repetition of orally presented words) and non-word visual stimuli (faces, objects, colors and numbers). Although the patient showed some impairments in naming faces, objects and colors, his performance scores were only slightly lower or not significantly different relative to those of the patients with intact LMFC-OTS. These data provide compelling evidence that the LMFC-OTS is not requisite for processing non-visual word stimuli, thus has selectivity for visual word processing. Functional properties: With tasks involving multiple levels and aspects of word processing, including Chinese character reading, phonological judgment, semantic judgment, identity judgment of abstract visual word representation, lexical decision, perceptual judgment of visual word appearance, and dictation, copying, voluntary writing, etc., we attempted to reveal the most critical dysfunction caused by damage in the LMFC-OTS, thus to clarify the most essential function of this region. Results showed that in addition to dysfunctions in Chinese character reading, phonological and semantic judgment, the patient with lesions at LMFC-OTS failed to judge correctly whether two characters (including compound and simple characters) with different surface features (e.g., different fonts, printed vs. handwritten vs. calligraphy styles, simplified characters vs. traditional characters, different orientations of strokes or whole characters) had the same abstract representation. The patient initially showed severe impairments in processing both simple characters and compound characters. He could only copy a compound character in a stroke-by-stroke manner, but not by character-by-character or even by radical-by-radical manners. During the recovery process, namely five months later, the patient could complete the abstract representation tasks of simple characters, but showed no improvement for compound characters. However, he then could copy compound characters in a radical-by-radical manner. Furthermore, it seems that the recovery of copying paralleled to that of judgment of abstract representation. These observations indicate that lesions of the LMFC-OTS in the pure alexic patients caused several damage in the ability of extracting the abstract representation from lower level units to higher level units, and the patient had especial difficulty to extract the abstract representation of whole character from its secondary units (e.g., radicals or single characters) and this ability was resistant to recover from impairment. Therefore, the LMFC-OTS appears to be responsible for the multilevel (particularly higher levels) abstract representations of visual word form. Successful extraction seems independent on access to phonological and semantic information, given the alexic patient showed severe impairments in reading aloud and semantic processing on simple characters while maintenance of intact judgment on their abstract representation. However, it is also possible that the interaction between the abstract representation and its related information e.g. phonological and semantic information was damaged as well in this patient. Taken together, we conclude that: 1) the LMFC-OTS is necessary for Chinese character processing, 2) it is selective for Chinese character processing, and 3) its critical function is to extract multiple levels of abstract representation of visual word and possibly to transmit it to phonological and semantic systems.

汉字阅读中的字形、字音、字义及频率效应的研究

Unlike alphabetic languages, Chinese language is ideographical writing system. Each Chinese character is single-syllable and usually has a direct meaning. So Chinese characters are a kind of valuable experimental material used for research on reading and comparisons of the reading mechanism of different language. In this paper, the normal persons and the patients with semantic dementia were respectively scheduled for two parts of experimental studies on the orthographic, phonologic, semantic and frequency effects of reading of Chinese characters. The Stroop-like character-picture interference experimental paradigm was used to investigate the orthographic, phonologic, semantic and frequency effects of Chinese characters on picture naming when they were presented with pictures to normal persons. The results indicated that the orthographic facilitation effect, phonologic facilitation effect, and semantic interference effect occurred at different SOA values. The orthographic and phonologic facilitation effects were independent. It was for the first time shown that the interaction between orthographic variable and semantic variable occurred when the high-frequency Chinese characters were read. Phonologic representation was activated quicker than semantic representation, by comparison of their SOA. Generally, it means that there is reading without meaning in Chinese character among the normal persons. The orthographic, phonologic, semantic, frequency and concrete effects of Chinese characters were further investigated among the dementia patients with DAT(dementia of Alzheimer's type disease) or CVA or both. They all have an impaired semantic memory. The results showed that patients with dementia could read the names of the pictures aloud while they could not name them or match them with a right character correctly. This is reading impairment without meaning in Chinese among the dementia patients. Meanwhile, they had a selective reading impairment and more LARC(a legitimate alternative reading of components) mistakes especially when reading low-frequency irregular, low-frequency inconsistent and abstract Chinese characters. With the patients' semantic impairment developed, their ability to read the pictures names would remain whereas their ability to read low-frequency irregular and low-frequency inconsistency Chinese characters was reduced. These results indicated that low-frequency irregular Chinese characters can be read correctly only when it is supported by their semantic information. Based on the above results of reading without meaning and of reading of low-frequency irregular Chinese characters supported by their semantic information, it is reasonable to suggest that at least two routes are involved in the process of reading Chinese characters. They are direct phonologic route and indirect semantic route; moreover, the two routes are independent.

隔区损毁对大鼠空间认知能力的影响及其在Morris迷宫中搜索策略的比较

Separate groups of rats with bilateral or unilateral lesions in septum were tested for acquisition and retention of the Morris water maze spatial cognitive task. Some of the animals in each lesion group received preoperative training in the task. Other animals in each group received no preoperative training. The results indicate that although both lesions lead to a spatial cognitive impairment in both the acquisition and retention of the task, the animals with bilateral lesions were more severely impaired than were the animals with unilateral, as indicated by quantitative measure. Searching strategies were used as an index to eximine the qualitative difference in the animals swim be havior, we found that the unilateral damaged animals still tend to use "mapping" strategies to solve the task as in the case of control groups but the accuracy is lower. The searching strategies used by the bilateral damaged animals showed complex patterns. The acquisition group with bilateral tend to use random and paratic strategies, however, the retention group with bilateral had a tendacy to use paratic and taxic strategies. The difference between searching strategies and its dynamic, change possibly suggest that other cognitive processing systems play an role in the processing of information about the task.

Aphasia rehabilitation and the strange neglect of speed.

Timing data is infrequently reported in aphasiological literature and time taken is only a minor factor, where it is considered at all, in existing aphasia assessments. This is not surprising because reaction times are difficult to obtain manually, but it is a pity, because speed data should be indispensable in assessing the severity of language processing disorders and in evaluating the effects of treatment. This paper argues that reporting accuracy data without discussing speed of performance gives an incomplete and potentially misleading picture of any cognitive function. Moreover, in deciding how to treat, when to continue treatment and when to cease therapy, clinicians should have regard to both parameters: Speed and accuracy of performance. Crerar, Ellis and Dean (1996) reported a study in which the written sentence comprehension of 14 long-term agrammatic subjects was assessed and treated using a computer-based microworld. Some statistically significant and durable treatment effects were obtained after a short amount of focused therapy. Only accuracy data were reported in that (already long) paper, and interestingly, although it has been a widely read study, neither referees nor subsequent readers seemed to miss "the other side of the coin": How these participants compared with controls for their speed of processing and what effect treatment had on speed. This paper considers both aspects of the data and presents a tentative way of combining treatment effects on both accuracy and speed of performance in a single indicator. Looking at rehabilitation this way gives us a rather different perspective on which individuals benefited most from the intervention. It also demonstrates that while some subjects are capable of utilising metalinguistic skills to achieve normal accuracy scores even many years post-stroke, there is little prospect of reducing the time taken to within the normal range. Without considering speed of processing, the extent of this residual functional impairment can be overlooked.

Exploitation of Ebola Virus VP35 Protein to identify new drugs counteracting its type I IFN antagonism

Ebolaviruses (EBOVs) are among the most virulent and deadly pathogens ever known, causing fulminant haemorrhagic fevers in humans and non-human primates. The 2014 outbreak of Ebola virus disease (EVD) in West Africa has claimed more lives than all previous EVD outbreaks combined. The EBOV high mortality rates have been related to the virus-induced impairment of the host innate immunity reaction due to two virus-coded proteins, VP24 and VP35. EBOV VP35 is a multifunctional protein, it is essential for viral replication as a component of the viral RNA polymerase and it also participates in nucleocapsid assembly. Early during EBOV infection, alpha-beta interferon (IFN-α/β) production would be triggered upon recognition of viral dsRNA products by cytoplasmic retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs). However, this recognition is efficiently prevented by the double-stranded RNA (dsRNA) binding activity of the EBOV VP35 protein, which hides RLRs binding sites on the dsRNA phosphate backbone as well the 5’-triphosphate (5’-ppp) dsRNA ends to RIG-I recognition. In addition to dsRNA binding and sequestration, EBOV VP35 inhibits IFN-α/β production preventing the activation of the IFN regulatory factor 3 (IRF-3) by direct interaction with cellular proteins. Previous studies demonstrated that single amino acid changes in the VP35 dsRNA binding domain reduce EBOV virulence, indicating that VP35 is an attractive target for antiviral drugs development. Within this context, here we report the establishment of a novel method to characterize the EBOV VP35 inhibitory function of the dsRNA-dependent RIG-I-mediated IFN-β signaling pathway in a BLS2 cell culture setting. In such system, a plasmid containing the promoter region of IFN-β gene linked with a luciferase reporter gene was transfected, together with a EBOV VP35 mammalian expression plasmid, into the IFN-sensitive A549 cell line, and the IFN-induction was stimulated through dsRNA transfection. Through alanine scanning mutational studies with biochemical, cellular and computational methods we highlighted the importance of some VP35 residues involved in dsRNA end-capping binding, such as R312, K282 and R322, that may serve as target for the development of small-molecule inhibitors against EBOV. Furthermore, we identified a synthetic compound that increased IFN-induction only under antiviral response stimulation and subverted VP35 inhibition, proving to be very attractive for the development of an antiviral drug. In conclusion, our results provide the establishment of a new assay as a straightforward tool for the screening of antiviral compounds that target i) dsRNA-VP35 or cellular protein-VP35 interaction and ii) dsRNA-dependent RIG-I-mediated IFN signaling pathway, in order to potentiate the IFN response against VP35 inhibition, setting the bases for further drug development.

Neuroprotective and immunomodulatory properties of the new PPARγ agonist MDG548: an in vitro and in vivo study in PD models

Neuroinflammation is a key component of Parkinson’s disease (PD) neuropathology. Skewed microglia activation with pro-inflammatory prevailing over anti-inflammatory phenotypes may contribute to neurotoxicity via the production of cytokines and neurotoxic species. Therefore, microglia polarization has been proposed as a target for neuroprotection. The peroxisome proliferator-activated receptor gamma (PPARγ) is expressed in microglia and peripheral immune cells, where it is involved in macrophages polarization and in the control of inflammatory responses, by modulating gene transcription. Several studies have shown that PPARγ agonists are neuroprotective in experimental PD models in rodents and primates. however safety concerns have been raised about PPARγ agonists thiazolidinediones (TZD) currently available, prompting for the development of non-TZD compounds. Aim of this study was to characterize a novel PPARγ agonist non TZD, MDG548, for its potential neuroprotective effect in PD models and its immunomodulatory activity as the underlying mechanism of neuroprotection. The neuroprotective activity of MDG548 was assessed in vivo in the subacute MPTP model and in the chronic MPTP/probenecid (MPTPp) model of PD. MDG548 activity on microglia activation and phenotype was investigated in the substantia nigra pars compacta (SNc) via the evaluation of pro- (TNF-α and iNOS) and anti-inflammatory (CD206) molecules, with fluorescent immunohistochemistry. Moreover, cultured murine microglia MMGT12 were treated with MDG548 in association with the inflammagen LPS, pro- and anti-inflammatory molecules were measured in the medium by ELISA assay and phagocytosis was evaluated by fluorescent immunohistochemistry for CD68. MDG548 arrested dopaminergic cells degeneration in the SNc in both the subacute MPTP and the chronic MPTPp models of PD, and reverted MPTPp-induced motor impairment. Moreover, MDG548 reduced microglia activation, iNOS and TNF-α production, while induced CD206 in microglia. In cultured unstimulated microglia, LPS increased TNF-α production and CD68 expression, while decreased CD206 expression. MDG548 reverted LPS effect on TNF-α and CD206 restoring physiological levels, while strongly increased CD68 expression. Results suggest that the PPARγ agonist MDG548 is neuroprotective in experimental models of PD. MDG548 targets microglia polarization by correcting the imbalance between pro- over antiinflammatory molecules, offering a novel immunomodulatory approach to neuroprotection.

The effect of concentration and duration of normobaric oxygen in reducing caspase-3 and -9 expression in a rat-model of focal cerebral ischaemia.

The aim of this study was to determine the effect of different concentrations of normobaric oxygen (NBO) on neurological function and the expression of caspase-3 and -9 in a rat model of acute cerebral ischaemia. Sprague-Dawley rats (n=120) were randomly divided into four groups (n=30 per group), including 3 groups given NBO at concentrations of 33%, 45% or 61% and one control group given air (21% oxygen). After 2 h of ischaemic occlusion, each group was further subdivided into six subgroups (n=5) during reperfusion according to the duration (3, 6, 12, 24, 48 or 72 h) and concentration of NBO (33%, 45% or 61%) or air treatment. The Fluorescence Quantitative polymerase chain reaction (PCR) and immunohistochemistry were used to detect caspase-3 and -9 mRNA and protein relative expression respectively. The Neurologic Impairment Score (NIS) was significantly lower in rats given 61% NBO ≥3 h after reperfusion when compared to the control group (P<0.05, Mann–Whitney U). NBO significantly reduced caspase-3 and -9 mRNA and protein expression when compared to the control group at all NBO concentrations and time points (P<0.05, ANOVA). The expression of caspase-3 and -9 was lower in the group given 61% NBO compared any other group, and this difference was statistically significant when compared to the group given 33% NBO for ≥48 h and the control group (both P<0.05, ANOVA). These findings indicate that NBO may inhibit the apoptotic pathway by reducing caspase-3 and -9 expression, thereby promoting neurological functional recovery after stroke.