Therapeutic effects of dietary intervention on neuroinflammation and brain metabolism in a rat model of photothrombotic stroke

INTRODUCTION: A possible target for stroke management is modulation of neuroinflammation. Evidence suggests that food components may exert anti-inflammatory properties and thus may reduce stroke-induced brain damage. AIM: To investigate the efficacy of a diet, containing anti-inflammatory ingredients, as treatment for focal ischemic brain damage induced by photothrombotic stroke in the somatosensory cortex of rats. RESULTS: Brain lesions were surrounded by strong astrogliosis on both day 7 and day 21 after stroke and were accompanied by a trend toward globally decreased glucose metabolism on day 7. The investigational diet applied 2 weeks before the ischemia did not affect astrocyte activation on day 7, but reduced it at day 21. The investigational diet applied immediately after the ischemia, increased astrocyte activation on day 7 and completely reversed this effect on day 21. Moreover, postischemic intervention increased glucose metabolism in somatosensory cortex ipsilateral to the lesion on day 7. CONCLUSION: This study reveals potentially beneficial effects of a diet containing elevated amounts of anti-inflammatory nutrients on the recovery from ischemic brain damage. Therefore, dietary intervention can be considered as an adjuvant therapy for recovery from this brain pathology

High Content Analysis of Hippocampal Neuron-Astrocyte Co-cultures Shows a Positive Effect of Fortasyn Connect on Neuronal Survival and Postsynaptic Maturation

Neuronal and synaptic membranes are composed of a phospholipid bilayer. Supplementation with dietary precursors for phospholipid synthesis -docosahexaenoic acid (DHA), uridine and choline- has been shown to increase neurite outgrowth and synaptogenesis bothin vivoandin vitro. A role for multi-nutrient intervention with specific precursors and cofactors has recently emerged in early Alzheimer's disease, which is characterized by decreased synapse numbers in the hippocampus. Moreover, the medical food Souvenaid, containing the specific nutrient combination Fortasyn Connect (FC), improves memory performance in early Alzheimer's disease patients, possibly via maintaining brain connectivity. This suggests an effect of FC on synapses, but the underlying cellular mechanism is not fully understood. Therefore, we investigated the effect of FC (consisting of DHA, eicosapentaenoic acid (EPA), uridine, choline, phospholipids, folic acid, vitamins B12, B6, C and E, and selenium), on synaptogenesis by supplementing it to primary neuron-astrocyte co-cultures, a cellular model that mimics metabolic dependencies in the brain. We measured neuronal developmental processes using high content screening in an automated manner, including neuronal survival, neurite morphology, as well as the formation and maturation of synapses. Here, we show that FC supplementation resulted in increased numbers of neurons without affecting astrocyte number. Furthermore, FC increased postsynaptic PSD95 levels in both immature and mature synapses. These findings suggest that supplementation with FC to neuron-astrocyte co-cultures increased both neuronal survival and the maturation of postsynaptic terminals, which might aid the functional interpretation of FC-based intervention strategies in neurological diseases characterized by neuronal loss and impaired synaptic functioning

The development of automated patch clamp assays for canonical transient receptor potential channels TRPC3, 6, and 7

The canonical transient receptor potential channel subfamily (TRPC3, TRPC6, and TRPC7) contains Ca2+ permeable non-selective cation channels that are widely expressed in a variety of tissues. There is increasing evidence implicating TRPC channels, particularly TRPC3 and 6, in physiological and pathophysiological processes, eliciting interest in these channels as novel drug targets. Electrophysiology remains a benchmark technique for measuring ion channel function and accurately determining the pharmacological effects of compounds. In this report we describe the development of TRPC inhibitor assays on 2 automated planar patch clamp platforms - the IonWorks® Quattro™ and QPatch® systems. To enable activation of TRPC channels by carbachol, Chinese Hamster Ovary-K1 cells stably expressing the muscarinic M3 receptor were transduced with human TRPC3, TRPC6, or TRPC7 using BacMam viruses. TRPC3, 6, and 7 currents could be recorded on both platforms. However, the design of each platform limits which assay parameters can be recorded. Due to its continuous recording capabilities, the QPatch can capture both the activation and decay of the response. However, the transient nature of TRPC channels, the inability to reactivate and the large variation in peak currents limits the ability to develop assays for compound screening. The IonWorks Quattro, due to its discontinuous sampling, did not fully capture the peak of TRPC currents. However, due to the ability of the IonWorks Quattro to record from 64 cells per well, the variation from well to well was sufficiently reduced allowing for the development of medium-throughput screening assays. © Copyright 2014, Mary Ann Liebert, Inc. 2014

Anti-inflammatory effects of rice bran components

Neuroinflammation has been implicated in the pathology of various psychiatric and neurodegenerative disorders. Accumulating evidence suggests that food components can modulate inflammatory processes, and therefore it could be hypothesized that such nutrients might exhibit therapeutic efficacy against these brain diseases. Rice bran is often discarded as a waste product, although it contains a wide range of potentially useful substances. Several rice fiber components from rice bran have been described as having antiinflammatory properties. This review summarizes the evidence supporting a modulatory effect of rice fiber components on symptoms in several animal models for neuroinflammation. In vitro studies on immune cells and in vivo studies on nutritional intervention in animal models of central and peripheral inflammation are discussed in the context of the potential use of rice fiber components for prevention and treatment of brain diseases in which neuroinflammation is involved

GPR39-expressing cells.

<p>(<b>A</b>) a bright field image of cobblestone-like cells. Scale bar = 100 µm. (<b>B</b>) <i>Gpr39</i> mRNA levels were evaluated by Taqman gene expression analysis in both muscle tissues and cultured cells. After 7 days in culture, <i>Gpr39</i> levels were up-regulated 163.9 fold in the cultures (n = 3). (<b>C</b>) Cells were loaded with Fura2-AM. Shown are ratiometric images of Ca²⁺ signals before and after 100 µM Zn²⁺ challenge. [Ca²⁺]<i>_i</i> is indicated on a rainbow scale with blue representing low [Ca²⁺]<i>_i</i> and orange/red high [Ca²⁺]<i>_i</i>. (<b>D</b>) Ca²⁺ signals were elicited in cells cultured from wild-type mice, but not <i>Gpr39</i>^−/− mice. (<b>E</b>) Zn²⁺ induces Ca²⁺ signals in Ca²⁺ free bath solution. (<b>F</b>) Cells were pre-incubated with or without 1 µM thapsigargin to deplete intracellular Ca²⁺ stores. Thapsigargin treatment not only abolished Zn²⁺ elicited Ca²⁺ signals, but also resulted in a decrease in [Ca²⁺]_i after Zn²⁺ application, the mechanism of which is still unknown.</p

GPR39 activation is functionally linked with TMEM16A channel opening.

<p>(<b>A</b>) Time courses for the current measured at +80 mV from individual cells. Both Ionomycin (1 µM) and Zn²⁺ evoked currents in FLCs from WT mice (n = 11, top trace), while Zn²⁺ was ineffective in <i>Gpr39</i>^−/− FLCs (n = 6, bottom trace). The data are summarized in panel (<b>B</b>). (<b>C</b>) Knockdown of <i>Tmem16a</i> by siRNA in FLCs were verified by Taqman PCR (94% reduction in <i>Tmem16a</i> expression), whereas <i>Gpr39</i> expression were unaffected (n = 3). (<b>D</b>) Zn²⁺-induced Ca²⁺ responses were measured in FLCs. No difference was observed between control and <i>Tmem16a</i> siRNA transfected cells (n = 54 and 67). In contrast, the size of Zn²⁺-induced current was reduced to about 32% in transfected cells (n = 7 and 12).</p

Zn²⁺ induces a large current in FLCs.

<p>(<b>A</b>) Membrane potentials were recorded in current clamp configuration. Zn²⁺ induced rapid membrane depolarization in FLCs cultured from wild-type mice, but not <i>Gpr39</i>^−/− cells. (<b>B</b>) Bar chart summarizing Zn²⁺ induced membrane depolarization in FLCs as in panel A. p<0.01. (<b>C</b> and <b>D</b>) Zn²⁺ induced a large current in wild-type. Current was sampled at +80 and −80 mV. (<b>C</b>) shows a time-series plot for a typical Zn²⁺-induced current in FLCs (n = 21). (<b>D</b>) shows the corresponding I-V relationship. (<b>E</b> and <b>F</b>) are similar experiments using FLCs isolated from GPR39 knockout mice and show that no Zn²⁺-induced current was observed in <i>Gpr39</i>^−/− FLCs (n = 11).</p

Data_Sheet_1_High phenylalanine concentrations induce demyelination and microglial activation in mouse cerebellar organotypic slices.pdf

Phenylketonuria (PKU) is an inborn error of metabolism. Mutations in the enzyme phenylalanine hydroxylase (PAH)-encoding gene lead to a decreased metabolism of the amino acid phenylalanine (Phe). The deficiency in PAH increases Phe levels in blood and brain. Accumulation of Phe can lead to delayed development, psychiatric problems and cognitive impairment. White matter (WM) damage is a neuropathological hallmark of PKU and can be seen even in early detected and treated PKU patients. The mechanisms linking high Phe concentrations to WM abnormalities remain unclear. We tested the effects of high Phe concentrations on myelin in three in vitro models of increasing complexity: two simple cell culture models and one model that preserves local brain tissue architecture, a cerebellar organotypic slice culture prepared from postnatal day (P) 8 CD-1 mice. Various Phe concentrations (0.1–10 mM) and durations of exposure were tested. We found no toxic effect of high Phe in the cell culture models. On the contrary, the treatment promoted the maturation of oligodendrocytes, particularly at the highest, non-physiological Phe concentrations. Exposure of cerebellar organotypic slices to 2.4 mM Phe for 21 days in vitro (DIV), but not 7 or 10 DIV, resulted in a significant decrease in myelin basic protein (MBP), calbindin-stained neurites, and neurites co-stained with MBP. Following exposure to a toxic concentration of Phe, a switch to the control medium for 7 days did not lead to remyelination, while very active remyelination was seen in slices following demyelination with lysolecithin. An enhanced number of microglia, displaying an activated type morphology, was seen after exposure of the slices to 2.4 mM Phe for 10 or 21 DIV. The results suggest that prolonged exposure to high Phe concentrations can induce microglial activation preceding significant disruption of myelin.</p

The development of automated patch clamp assays for canonical transient receptor potential channels TRPC3, 6, and 7

The canonical transient receptor potential channel subfamily (TRPC3, TRPC6, and TRPC7) contains Ca2+ permeable non-selective cation channels that are widely expressed in a variety of tissues. There is increasing evidence implicating TRPC channels, particularly TRPC3 and 6, in physiological and pathophysiological processes, eliciting interest in these channels as novel drug targets. Electrophysiology remains a benchmark technique for measuring ion channel function and accurately determining the pharmacological effects of compounds. In this report we describe the development of TRPC inhibitor assays on 2 automated planar patch clamp platforms - the IonWorks® Quattro™ and QPatch® systems. To enable activation of TRPC channels by carbachol, Chinese Hamster Ovary-K1 cells stably expressing the muscarinic M3 receptor were transduced with human TRPC3, TRPC6, or TRPC7 using BacMam viruses. TRPC3, 6, and 7 currents could be recorded on both platforms. However, the design of each platform limits which assay parameters can be recorded. Due to its continuous recording capabilities, the QPatch can capture both the activation and decay of the response. However, the transient nature of TRPC channels, the inability to reactivate and the large variation in peak currents limits the ability to develop assays for compound screening. The IonWorks Quattro, due to its discontinuous sampling, did not fully capture the peak of TRPC currents. However, due to the ability of the IonWorks Quattro to record from 64 cells per well, the variation from well to well was sufficiently reduced allowing for the development of medium-throughput screening assays. © Copyright 2014, Mary Ann Liebert, Inc. 2014

The EAT–Lancet reference diet and cognitive function across the life course

The EAT–Lancet Commission devised a sustainable reference diet with the aim of reducing the incidence of non-communicable diseases and mortality globally while improving food system sustainability. The extent to which the reference diet supports cognitive function across the life course, however, has not yet been evaluated. This Review assesses the evidence for diet supporting cognitive function from childhood into old age. A comprehensive but non-exhaustive literature search was done, synthesising studies that investigated the effect of whole foods on cognition in healthy, community-dwelling human participants. We found that the current evidence base is weak with mixed conclusions and multiple methodological caveats, which precludes strong conclusions pertaining to the suitability of dietary recommendations for each food group per age group. Long-term intervention and prospective cohort studies are needed to reduce this knowledge deficit. Revising dietary recommendations with the aim of maintaining an adequate nutrient intake to sustain healthy cognitive function across the life course could be worthwhile. This Review outlines recommendations for future work to help improve the current knowledge deficit regarding dietary intake and cognitive function across the life course and its implications for dietary guidelines such as the EAT–Lancet Commission