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Aim: the aim of the study was to determine how the addition of a DNA oligonucleotide cargo to 3-nm gold glyconanoparticles would affect tissue distribution. Methods: gold glyconanoparticles with 1-6 covalently bound oligonucleotides (40 nt dsDNA) were injected into rats and allowed to circulate for 10 min. Organs were harvested and gold quantitated by inductively coupled plasma mass spectrometry. Cellular localization of the nanocarriers was determined by electron microscopy. Results and conclusion: addition of DNA cargo to the nanocarriers prevented localization in the kidney but increased localization in liver hepatocytes and splenic macrophages. There was no significant change in heart, lung or brain. DNA increases the size and adds a strong negative charge to the nanoparticles, which radically affects tissue distribution., Sheffield Hospitals Trust

Most currently available antiepileptics are not fully effective in the prevention of seizures in absence epilepsy owing to the presence of blood-brain barrier (BBB). We aimed to test whether binding an antiepileptic drug, lacosamide (LCM), to glucose-coated gold nanoparticles (GNPs) enables efficient brain drug delivery to suppress the epileptic activity in WAG/Rij rats with absence epilepsy.In these animals, intracranial-EEG recording, behavioral test, in vivo imaging of LCM and LCM-GNP conjugate distribution in the brain, inductively coupled plasma mass spectrometry analysis, immunofluorescence staining of glucose transporter (Glut)- 1, glial fibrillary acidic protein (GFAP), and p-glycoprotein (P-gp) and electron microscopy were performed.Lacosamide-GNP conjugates decreased the amplitude and frequency of spike-wave-like discharges (SWDs) and alleviated the anxiety-like behavior as assessed by EEG and elevated plus-maze test, respectively (p 0.01). The in vivo imaging system results showed higher levels of fluorescein dye tagged to LCM-GNP in the brain during the 5-day injection period (p 0.01). Immunofluorescence staining displayed decreased P-gp, Glut-1, and GFAP expression by LCM-GNP conjugate treatment predominantly in the cerebral cortex suggesting a potential functionality of this brain region in the modulation of neuronal activity in our experimental setting (p 0.01).We suggest that the conjugation of LCM to GNPs may provide a novel approach for efficient brain drug delivery in light of the effectiveness of our strategy not only in suppressing the seizure activity but also in decreasing the need to use high dosages of the antiepileptics to reduce the frequently encountered side effects in drug-resistant epilepsy.

ObjectiveBipolar disorder (BD) patients have recently been shown to exhibit increased proinflammatory cytokine levels indicating the role of inflammation in this disease. As inflammatory responses often include complement level alterations and complement production is influenced by cytokines, we aimed to find out whether complement system is activated in BD in a time-dependent manner and complement factors are involved in BD pathogenesis.MethodsSerum C4, factor B, sC5b-9 and neuron-specific enolase levels were measured by enzyme-linked immunosorbent assay, whereas peripheral blood mononuclear cell messenger RNA (mRNA) expression levels of C1q, C4, factor B and CD55 were measured by real-time polymerase chain reaction in chronic BD patients (n=22), first episode BD patients (n=24) and healthy controls (n=19).ResultsSerum complement levels were significantly reduced in chronic BD patients as compared with first episode BD patients and healthy controls. Serum levels of complement factors showed significant inverse correlation with disease duration, severity of manic symptoms and serum neuron-specific enolase levels. In chronic BD patients, peripheral blood mononuclear cell mRNA expression levels of C1q, C4 and factor B were significantly elevated, whereas the mRNA expression level of the complement inhibitor CD55 was significantly reduced.ConclusionsOur results suggest that complement factor levels are reduced in BD presumably due to overconsumption of the complement system and complement production is increased at mRNA level possibly as a compensation measure. Complement factors might potentially be used as indicators of disease severity, neuronal loss and cognitive dysfunction.

The increase in the circulatory cytokine levels observed in patients with bipolar disorder (BD) may imply involvement of inflammation in the pathogenesis of mood disorders. However, the association between the inflammatory process and the stage and severity of illness is not well understood. In this study, our aim was to investigate the association between neuroinflammation and disease progression in the clinical course of BD.IL-6, TNF-α, IL-1 receptor antagonist (IL-1RA), neuronspecific enolase (NSE) and S100B were measured by ELISA in plasma samples of patients at early-stage BD (n=30), chronic BD (n=77) and healthy controls (n=30).Chronic BD patients showed significantly increased levels of all measured inflammatory markers as compared to early-stage BD patients and the healthy controls. IL-6 and IL-1RA levels correlated with NSE and/or S100B levels and TNF-α levels correlated with Montgomery- Asberg Depression Rating Scale scores and Clinical Global Impression Scale scores.Our results indicate that inflammation appears to be particularly associated with IL-1RA and IL-6 activity, progressing at later stages of BD and possibly associated with gliosis and neuronal loss.

To investigate the role of metabotrophic purinergic P2Y receptors in neuroblastoma cell survival, expression of P2 receptors by normal mouse (C57BL/6) brain and human neuroblastoma SH-SY5Y cells was investigated by Western blot and real time PCR studies. Viability of SH-SY5Y cells treated with purinergic receptor antagonists suramin and pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS) was evaluated by MTT assay and flow cytometry. In the brain samples of C57BL/6 mice, expressions of P2Y4 and P2X7 were significantly reduced, whereas that of P2Y1 was significantly elevated in an age-dependent manner. SH-SY5Y cell viability was significantly reduced and necrotic cell rates were mildly increased by 400 μM suramin and 100 μM PPADS treatment. Antagonist treatment downregulated P2Y1, P2Y2 and P2Y4 and upregulated P2Y6, P2Y12 and P2X7 mRNA levels in SH-SY5Y cells on the 24th hour. These alterations were abolished for all P2 receptors except P2Y1 in the 48th hour. P2Y receptors are expressed by both normal mouse brain and human neuroblastoma cells. Purinergic receptor antagonism interferes with neuroblastoma viability through elevation of necrotic cell death and modulation of P2 receptor expression. P2Y receptors might thus be useful targets for future anti-tumor treatment trials.

Objective The increase in the circulatory cytokine levels observed in patients with bipolar disorder (BD) may imply involvement of inflammation in the pathogenesis of mood disorders. However, the association between the inflammatory process and the stage and severity of illness is not well understood. In this study, our aim was to investigate the association between neuroinflammation and disease progression in the clinical course of BD. Method IL-6, TNF-α, IL-1 receptor antagonist (IL-1RA), neuronspecific enolase (NSE) and S100B were measured by ELISA in plasma samples of patients at early-stage BD (n=30), chronic BD (n=77) and healthy controls (n=30). Results Chronic BD patients showed significantly increased levels of all measured inflammatory markers as compared to early-stage BD patients and the healthy controls. IL-6 and IL-1RA levels correlated with NSE and/or S100B levels and TNF-α levels correlated with Montgomery- Asberg Depression Rating Scale scores and Clinical Global Impression Scale scores. Conclusion Our results indicate that inflammation appears to be particularly associated with IL-1RA and IL-6 activity, progressing at later stages of BD and possibly associated with gliosis and neuronal loss.

The blood-brain barrier (BBB) permeability primarily increases in cerebral venules during acute hypertension. Methyl-beta-cyclodextrin (M beta CD), a cholesterol-depleting agent, decreases the expression of caveolins disrupting caveolar structures. We aimed to determine the effects of M beta CD on the BBB permeability of angiotensin (ANG) II-induced hypertensive rats. Three minutes after M beta CD administration (5 mg/kg), acute hypertension was induced by ANG II (60 mu g/kg). Evans blue (EB) and horseradish peroxidase (HRP) tracers were used to assess BBB permeability. Immunohistochemistry for caveolin (Cav)-1 and tight junction protein claudin-5 was performed. EB dye content significantly increased in both cerebral cortices and left hippocampus in M beta CD (P < 0.05), right cerebral cortex and both hippocampi in ANG II (P < 0.05; P < 0.01), and both cerebral cortices and hippocampi in M beta CD plus ANG II groups compared with controls (P < 0.05; P < 0.01). A significant decrease in claudin-5 immunostaining intensity was observed in animals treated with M beta CD compared with controls (P < 0.05). Cav-1 immunostaining intensity increased in ANG II group (P < 0.05). Ultrastructurally, HRP reaction products were observed in endothelial cells of the microvessels in the hippocampus region in M beta CD group while the tracer was mainly localized in astrocytes and neurons in ANG II, and M beta CD plus ANG II groups. The endothelial cells of the venules in the cerebral cortex of the animals in the latter experimental groups also showed an abundance of caveolar vesicles devoid of HRP reaction products. Our results revealed that M beta CD did not provide overall protective effects on BBB integrity in acute hypertension and even led to BBB disruption in normotensive animals.

Background/Aim: Obstructive sleep apnea syndrome (OSAS) is associated with intermittent episodes of hypoxia, endothelial dysfunction and associated cardiovascular problems. Our aim was to investigate whether OSAS-related hypoxia alters the expression of rho-associated protein kinase (ROCK), a marker of chronic hypoxia and endothelial dysfunction. Materials and Methods: ROCK1 and ROCK2 levels were measured by immunoblotting in peripheral blood mononuclear cells (PBMC) of 47 OSAS patients and 17 healthy controls. Results: OSAS patients showed significantly lower PBMC ROCK1 and ROCK2 levels than healthy controls in the morning, but not in the evening. ROCK1/2 levels were correlated with blood triglyceride, visceral adiposity index, minimum oxygen saturation, C-reactive protein concentration, lymphocyte levels and sleep efficiency. Conclusion: Intermittent hypoxia induced by OSAS does not permanently alter ROCK protein expression levels. OSAS appears to be associated with endothelial dysfunction through inflammation and lipid metabolism pathways.

Background Intravenous (IV) immunoglobulin (Ig) treatment is known to alleviate behavioral deficits and increase survival in the experimentally induced model of sepsis. To delineate the mechanisms by which IVIg treatment prevents neuronal dysfunction, an array of immunological and apoptosis markers was investigated. Methods Sepsis was induced by cecal ligation perforation (CLP) in rats. The animals were divided into five groups: sham, control, CLP + saline, CLP + immunoglobulin G (IgG) (250 mg/kg, iv), and CLP + immunoglobulins enriched with immunoglobulin M (IgGAM) (250 mg/kg, iv). Blood and brain samples were taken in two sets of experiments to see the early (24 h) and late (10 days) effects of treatment. Total complement activity, complement 3 (C3), and soluble complement C5b-9 levels were measured in the sera of rats using ELISA-based methods. Cerebral complement, complement receptor, NF-κB, Bax, and Bcl-2 expressions were analyzed by western blot and/or RT-PCR methods. Immune cell infiltration and gliosis were examined by immunohistochemistry using CD3, CD4, CD8, CD11b, CD19, and glial fibrillary acidic protein antibodies. Apoptotic neuronal death was investigated by TUNEL staining. Results IVIgG and IgGAM administration significantly reduced systemic complement activity and cerebral C5a and C5a receptor expression. Likewise, both treatment methods reduced proapoptotic NF-κB and Bax expressions in the brain. IVIgG and IgGAM treatment induced considerable amelioration in glial cell proliferation and neuronal apoptosis which were increased in non-treated septic rats. Conclusions We suggest that IVIgG and IgGAM administration ameliorates neuronal dysfunction and behavioral deficits by reducing apoptotic cell death and glial cell proliferation. In both treatment methods, these beneficial effects might be mediated through reduction of anaphylatoxic C5a activity and subsequent inhibition of inflammation and apoptosis pathways. Electronic supplementary material The online version of this article (doi:10.1186/s40635-016-0114-1) contains supplementary material, which is available to authorized users.

IntroductionThere is growing evidence about neuroinflammation in the aetiopathogenesis of bipolar disorder. Early diagnosis and intervention strategies are thought to be excessively important lately.ObjectivesTo check neuroinflammation levels in early stage bipolar disorder and explore the associations with clinical variables.AimsWe aimed to evaluate inflammation and neurodegeneration findings in early stage bipolar disorder.MethodsSerum interleukin 1-receptor antagonist (IL-1Ra), interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-α), high sensitive C reactive protein (hs-CRP), S100B and neuron specific enolase (NSE) levels were assessed by enzyme-linked immunosorbent assays in a total of 30 patients with bipolar disorder in the early stage and compared with 30 matched healthy controls. The clinical symptoms were rated using Montgomery Asberg Depression Scale, Young Mania Rating Scale, Positive and Negative Syndrome Scale and Clinical Global Impression Scale.ResultsAmong the patients with bipolar disorder, 14 (% 46.6) were in a manic/hypomanic state and 12 (% 40) were in a euthymic state. Serum IL-6 levels were significantly higher (P = 0.018), TNF-α and S100B levels were significantly lower in the early stage group (P < 0.001 and P = 0.03, respectively). After repeated analysis with only drug-naive patients, the results showed no difference. There was a positive and significant correlation between TNF-α levels and CGI, MADRS scores (all P < 0.05); NSE levels and MADRS scores (P < 0.05).ConclusionsThis study supported the association of early stage bipolar disorder with inflammation and neurodegeneration. IL-6 may be a potential biomarker. Thus, early diagnosis and intervention may be crucial to prevent progressive neuroinflammation and neurodegeneration in early stages of disorder.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by progressive degeneration of the dopaminergic neurons in substantia nigra, presumably due to increased apoptosis and oxidative stress. To investigate whether PD-induced survival/apoptosis gene expression changes can serve as prognostic biomarkers of PD, we measured expression levels of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/Akt pathway factors and additional apoptotic and anti-apoptotic factors in peripheral blood mononuclear cells (PBMC) of PD patients (n=50) and healthy controls (n=50) by real time PCR. Expression levels of apoptotic factors phosphatase and tensin homolog (PTEN) and mitochondrial apoptosis-inducing factor 1 (AIFM1) were significantly decreased, anti-apoptotic factors DJ-1 and Akt-1 were significantly increased and anti-apoptotic Bcl-2 was significantly decreased in PD patients. Expression levels of AIFM1 were significantly correlated with Hoehn-Yahr scores. Moreover, PD patients with postural instability showed significantly reduced expression levels of anti-apoptotic DJ-1, Akt-1 and mTOR than PD patients without postural instability. Expression profiles of brain samples of mice with rotenone-induced PD model and PBMC samples of PD patients showed remarkable resemblance. Our results indicate that the anti-apoptotic PI3K/Akt pathway is over activated in PD, presumably as an effort to compensate for increased neuronal apoptosis and oxidative stress. By contrast, patients with postural instability show reduced anti-apoptotic factor expression suggesting that this compensating mechanism fails in patients with this particular motor symptom. PBMC expression levels of AIFM1 might serve as a biomarker of disability and disease progression in PD.