A case-control comparison of acute-phase peripheral blood gene expression in participants diagnosed with minor ischaemic stroke or stroke mimics.

BACKGROUND: Past studies suggest that there are changes in peripheral blood cell gene expression in response to ischaemic stroke; however, the specific changes which occur during the acute phase are poorly characterised. The current study aimed to identify peripheral blood cell genes specifically associated with the early response to ischaemic stroke using whole blood samples collected from participants diagnosed with ischaemic stroke (n = 29) or stroke mimics (n = 27) following emergency presentation to hospital. Long non-coding RNA (lncRNA), mRNA and micro-RNA (miRNA) abundance was measured by RNA-seq, and the consensusDE package was used to identify genes which were differentially expressed between groups. A sensitivity analysis excluding two participants with metastatic disease was also conducted. RESULTS: The mean time from symptom onset to blood collection was 2.6 h. Most strokes were mild (median NIH stroke scale score 2.0). Ten mRNAs (all down-regulated in samples provided by patients experiencing ischaemic stroke) and 30 miRNAs (14 over-expressed and 16 under-expressed in participants with ischaemic stroke) were significantly different between groups in the whole cohort and sensitivity analyses. No significant over-representation of gene ontology categories by the differentially expressed genes was observed. Random forest analysis suggested a panel of differentially expressed genes (ADGRG7 and miRNAs 96, 532, 6766, 6798 and 6804) as potential ischaemic stroke biomarkers, although modelling analyses demonstrated that these genes had poor diagnostic performance. CONCLUSIONS: This study provides evidence suggesting that the early response to minor ischaemic stroke is predominantly reflected by changes in the expression of miRNAs in peripheral blood cells. Further work in independent cohorts particularly in patients with more severe stroke is needed to validate these findings and investigate their clinical relevance.

Serum angiopoietin-1 concentration does not distinguish patients with ischaemic stroke from those presenting to hospital with ischaemic stroke mimics.

BACKGROUND: A previous study found that circulating angiopoietin-1 (angpt-1) concentrations were significantly lower in patients who had a recent ischaemic stroke compared to healthy controls. The primary aim of this study was to assess whether serum angpt-1 could be used as a diagnostic test of ischemic stroke in patients presenting to hospital as an emergency. Exploratory analyses investigated the association of proteins functionally related to angpt-1 (angpt-2, Tie-2, matrix metalloproteinase-9 and vascular endothelial growth factors A, C and D) with ischaemic stroke diagnosis. METHODS: Patients presenting to Townsville University Hospital for emergency assessment of stroke-like symptoms were consecutively recruited and provided a blood sample. After assessment by a consultant neurologist, patients were grouped into those who did, or did not have ischaemic stroke. The potential for serum angpt-1 to diagnose ischaemic stroke was assessed using receiver operator characteristic (ROC) curves. Cross-sectional analyses appraised inter-group differences in the serum concentration of other proteins. RESULTS: One-hundred and twenty-six patients presenting to Townsville University Hospital for emergency assessment of stroke-like symptoms were recruited (median time from symptom onset to hospital presentation: 2.6 (inter-quartile range: 1.2-4.6) hours). Serum angpt-1 had poor ability to diagnose ischaemic stroke in analyses using the whole cohort, or in sensitivity analyses (area under the ROC curve 0.51 (95% CI: 0.41-0.62) and 0.52 (95% CI: 0.39-0.64), respectively). No associations of serum angpt-1 concentration with ischaemic stroke severity, symptom duration or aetiology were observed. Serum concentrations of the other assessed proteins did not differ between patient groups. CONCLUSIONS: Serum angpt-1 concentration is unlikely to be useful for emergency diagnosis of ischaemic stroke.

The Therapeutic Potential of Ketogenic Diet Throughout Life: Focus on Metabolic, Neurodevelopmental and Neurodegenerative Disorders.

This chapter reviews the efficacy of the ketogenic diet in a variety of neurodegenerative, neurodevelopmental and metabolic conditions throughout different stages of life. It describes conditions affecting children, metabolic disorders in adults and disorderrs affecting the elderly. We have focused on application of the ketogenic diet in clinical studies and in preclinical models and discuss the benefits and negative aspects of the diet. Finally, we highlight the need for further research in this area with a view of discovering novel mechanistic targets of the ketogenic diet, as a means of maximising the potential benefits/risks ratio.

Chronic Helminth Infection Perturbs the Gut-Brain Axis, Promotes Neuropathology, and Alters Behavior.

Helminth infections in children are associated with impaired cognitive development; however, the biological mechanisms for this remain unclear. Using a murine model of gastrointestinal helminth infection, we demonstrate that early-life exposure to helminths promotes local and systemic inflammatory responses and transient changes in the gastrointestinal microbiome. Behavioral and cognitive analyses performed 9-months postinfection revealed deficits in spatial recognition memory and an anxiety-like behavioral phenotype in worm-infected mice, which was associated with neuropathology and increased microglial activation within the brain. This study demonstrates a previously unrecognized mechanism through which helminth infections may influence cognitive function, via perturbations in the gut-immune-brain axis.

Ketogenic diet has a positive association with mental and emotional well-being in the general population.

OBJECTIVES: A ketogenic diet reduces pathologic stress and improves mood in neurodegenerative and neurodevelopmental disorders. However, the effects of a ketogenic diet for people from the general population have largely been unexplored. A ketogenic diet is increasingly used for weight loss. Research in healthy individuals primarily focuses on the physical implications of a ketogenic diet. It is important to understand the holistic effects of a ketogenic diet, not only the physiological but also the psychological effects, in non-clinical samples. The aim of this cross-sectional study with multiple cohorts was to investigate the association of a ketogenic diet with different aspects of mental health, including calmness, contentedness, alertness, cognitive and emotional stress, depression, anxiety, and loneliness, in a general healthy population. METHODS: Two online surveys were distributed: cohort 1 used Bond-Lader visual analog scales and Perceived Stress Scale (n = 147) and cohort 2 the Depression Anxiety Stress Scale and revised UCLA Loneliness Scale (n = 276). RESULTS: A ketogenic diet was associated with higher self-reported mental and emotional well-being behaviors, including calmness, contentedness, alertness, cognitive and emotional stress, depression, anxiety, and loneliness, compared with individuals on a non-specific diet in a general population. CONCLUSION: This research found that a ketogenic diet has potential psychological benefits in the general population.

Positive and negative schizotypy personality traits are lower in individuals on ketogenic diet in a non-clinical sample.

Schizotypal personality comprises traits such as odd beliefs, perceptual abnormalities, and social difficulties; these traits are distributed throughout the general population. While not meeting the clinical threshold for schizophrenia or schizotypal personality disorder, schizotypal personality traits still provide insights for understanding early clinical risk factors. Ketogenic diet reportedly reduces psychotic symptoms in preclinical and clinical studies. Therefore, we investigated whether ketogenic diet is associated with lower schizotypal traits in the general population. Participants following a ketogenic or other diet were recruited using opportunity sampling. Individuals completed a survey investigating general demographic, socioeconomic, health, diet and lifestyle questions, followed by the Schizotypal Personality Questionnaire - Brief Revised version (SPQ-BR). We found that individuals following a ketogenic diet (n = 118) had lower ideas of reference, magical thinking, suspiciousness, unusual perceptions, constricted affect, social anxiety scores, cognitive (positive) perceptual scores, interpersonal (negative) scores and total SPQ-BR compared to individuals on the other diets (n = 139). Magical thinking, constricted affect, social anxiety, cognitive perceptual, interpersonal scores and total SPQ-BR scores remained significant when we controlled for body mass index (BMI) and age. Disorganised features were not influenced by ketogenic diet. The longer individuals adhered to a ketogenic diet the lower their positive and negative schizotypy traits. These findings highlight that ketogenic diet is associated with lower non-clinical schizotypal personality traits. Our results suggest that ketogenic diet might have potential prophylactic properties for individuals at-risk for psychosis.

The use of integrated behavioural z-scoring in behavioural neuroscience - A perspective article.

Complex pathophysiology in psychiatric disorders results in difficulties interpreting pre-clinical data. Guilloux et al. (2011b), proposed an integrated behavioural z-scoring procedure to improve the predictive validity of animal models by converging evidence similarly used to diagnose mental health conditions in humans. Here, I set out to give a brief review of the current methodology and literature using integrated behavioural z-scoring. Secondly, I will discuss the benefits and downfalls of integrated behavioural z-scoring and its potential future applications. Integrated behavioural z-scoring is a methodology used most frequently within animal models of depression and anxiety. Here, I am suggesting broadening the application of integrated behavioural z-scoring beyond the field of depression and anxiety to a three-step methodology to obtain disease-specific behavioural z-scores (i.e Schizophrenia index, Alzheimer's disease index) to aid translatability and interpretation of data. Lastly, I suggest integrating not only behaviour but also biological variables to create converging psychological and physiological evidence to sustain face and construct validity, while improving predict validity.

Human nail cortisol as a retrospective biomarker of chronic stress: A systematic review.

Cortisol is the primary glucocorticoid produced by the activation of the hypothalamic pituitary adrenal (HPA) axis after a psychological or physiological stressor. The dysregulation of the HPA axis by chronic stress has been associated with psychiatric disorders. Although hair is currently the main validated source of chronic cortisol concentrations, cortisol is also bound to human nails, another keratinised matrix. Therefore, nail cortisol has the potential to be an alternative retrospective chronic measure of HPA activation. The aim of this systematic review was to assess the temporal resolution, methodological issues, HPA correlates, and target populations in nail cortisol investigations. A qualitative synthesis was performed to assess current literature exploring cortisol concentrations from human nails. A total of 18 eligible human studies extracted from Medline (PubMed and Ovid), ProQuest (PsycINFO), and Scopus found that immunoassays and mass spectrometry were the two primarily methods of analysis. However, methodological variability remained evident between studies. Nail cortisol correlated with saliva and hair in some studies and was investigated across multiple developmental periods. Finally, when applied as an outcome measure in health disorders, higher nail cortisol concentrations have been shown to be associated with acute coronary syndrome and depression. In conclusion, nail cortisol may serve as a retrospective biomarker of chronic stress; however, the ability to track how much cortisol is accumulating within nail clippings is complex and may represent a large timespan. Further, very few studies have reported effect sizes and investigated the effects of covariates, such as age, sex, ethnicity, and nail characteristics, which limits the validation of this measure. Further studies are required to validate the utility of nail cortisol as a biomarker of chronic stress across the human lifespan.

The Gut Microbiome in Psychosis From Mice to Men: A Systematic Review of Preclinical and Clinical Studies.

The gut microbiome is rapidly becoming the focus of interest as a possible factor involved in the pathophysiology of neuropsychiatric disorders. Recent understanding of the pathophysiology of schizophrenia emphasizes the role of systemic components, including immune/inflammatory and metabolic processes, which are influenced by and interacting with the gut microbiome. Here we systematically review the current literature on the gut microbiome in schizophrenia-spectrum disorders and in their animal models. We found that the gut microbiome is altered in psychosis compared to healthy controls. Furthermore, we identified potential factors related to psychosis, which may contribute to the gut microbiome alterations. However, further research is needed to establish the disease-specificity and potential causal relationships between changes of the microbiome and disease pathophysiology. This can open up the possibility of. manipulating the gut microbiome for improved symptom control and for the development of novel therapeutic approaches in schizophrenia and related psychotic disorders.

Ketogenic therapy in neurodegenerative and psychiatric disorders: From mice to men.

Ketogenic diet is a low carbohydrate and high fat diet that has been used for over 100 years in the management of childhood refractory epilepsy. More recently, ketogenic diet has been investigated for a number of metabolic, neurodegenerative and neurodevelopmental disorders. In this comprehensive review, we critically examine the potential therapeutic benefits of ketogenic diet and ketogenic agents on neurodegenerative and psychiatric disorders in humans and translationally valid animal models. The preclinical literature provides strong support for the efficacy of ketogenic diet in a variety of diverse animal models of neuropsychiatric disorders. However, the evidence from clinical studies, while encouraging, particularly in Alzheimer's disease, psychotic and autism spectrum disorders, is limited to case studies and small pilot trials. Firm conclusion on the efficacy of ketogenic diet in psychiatric disorders cannot be drawn due to the lack of randomised, controlled clinical trials. The potential mechanisms of action of ketogenic therapy in these disorders with diverse pathophysiology may include energy metabolism, oxidative stress and immune/inflammatory processes. In conclusion, while ketogenic diet and ketogenic substances hold promise pre-clinically in a variety of neurodegenerative and psychiatric disorders, further studies, particularly randomised controlled clinical trials, are warranted to better understand their clinical efficacy and potential side effects.

Protocol for the Use of the Ketogenic Diet in Preclinical and Clinical Practice.

Many age-related diseases are associated with metabolic abnormalities, and dietary interventions may have some benefit in alleviating symptoms or in delaying disease onset. Here, we review the commonly used best practices involved in applications of the ketogenic diet to facilitate its translation into clinical use. The findings reveal that better education of physicians is essential for applying the optimum diet and monitoring its effects in clinical practice. In addition, investigators should carefully consider potential confounding factors prior to commencing studies involving a ketogenic diet. Most importantly, current studies should improve their reporting on ketone levels as well as on the intake of both macro- and micronutrients. Finally, more detailed studies on the mechanism of action are necessary to help identify potential biomarkers for response prediction and monitoring, and to uncover new drug targets to aid the development of novel treatments.

Effects of beta-hydroxybutyrate administration on MK-801-induced schizophrenia-like behaviour in mice.

RATIONALE: Impaired cerebral glucose metabolism is a core pathological feature of schizophrenia. We recently demonstrated that a ketogenic diet, causing a shift from glycolysis to ketosis, normalized schizophrenia-like behaviours in an acute N-methyl-D-aspartate (NMDA) receptor antagonist model of the illness. Ketogenic diet produces the ketone body, ß-hydroxybutyrate (BHB), which may serve as an alternative fuel source in its own right without a strict dietary regime. OBJECTIVE: We hypothesized that chronic administration of BHB replicates the therapeutic effects of ketogenic diet in an acute NMDA receptor hypofunction model of schizophrenia in mice. METHODS: C57Bl/6 mice were either treated with acute doses of 2 mmol/kg, 10 mmol/kg, or 20 mmol/kg BHB or received daily intraperitoneal injections of 2 mmol/kg BHB or saline for 3 weeks. Behavioural testing assessed the effect of acute challenge with 0.2 mg/kg MK-801 or saline on open field behaviour, social interaction, and prepulse inhibition of startle (PPI). RESULTS: Acute BHB administration dose-dependently increased BHB plasma levels, whereas the 2 mmol/kg dose increased plasma glucose levels. The highest acute dose of BHB supressed spontaneous locomotor activity, MK-801-induced locomotor hyperactivity and MK-801-induced disruption of PPI. Chronic BHB treatment normalized MK-801-induced hyperlocomotion, reduction of sociability, and disruption of PPI. CONCLUSION: In conclusion, BHB may present a novel treatment option for patients with schizophrenia by providing an alternative fuel source to normalize impaired glucose metabolism in the brain.

Ketogenic diet prevents impaired prepulse inhibition of startle in an acute NMDA receptor hypofunction model of schizophrenia.

Recent transcriptomic, proteomic and metabolomics studies have highlighted an abnormal cerebral glucose and energy metabolism as one of the potential pathophysiological mechanisms of schizophrenia. This raises the possibility that a metabolically-based intervention might have therapeutic value in the management of schizophrenia, a notion supported by our recent results that a low carbohydrate/high-fat therapeutic ketogenic diet (KD) prevented a variety of behavioural abnormalities induced by pharmacological inhibition of NMDA glutamate receptors. Here we asked if the beneficial effects of KD can be generalised to impaired prepulse inhibition of startle (PPI), a translationally validated endophenotype of schizophrenia, in a pharmacological model in mice. Furthermore, we addressed the issue of whether the effect of KD is linked to the calorie-restricted state typical of the initial phase of KD. We fed male C57BL/6 mice a KD for 7 weeks and tested PPI at 3 and 7 weeks, in the presence and absence of a significant digestible energy deficit, respectively. We used an NMDA receptor hypo-function model of schizophrenia induced by acute injection of dizocilpine (MK-801). We found that KD effectively prevented MK-801-induced PPI impairments at both 3 and 7 weeks, irrespective of the presence or absence of digestible energy deficit. Furthermore, there was a lack of correlation between PPI and body weight changes. These results support the efficacy of the therapeutic KD in a translational model of schizophrenia and furthermore provide evidence against the role of calorie restriction in its mechanism of action.

The Elevated Plus Maze Test for Measuring Anxiety-Like Behavior in Rodents.

The elevated plus maze test is used to measure anxiety-like behavior in rodents. It can be used to gain insight into conditions such as posttraumatic stress disorder (PTSD) and other conditions marked by anxious behavior. It can also be used as a component in screening of novel compounds for anxiolytic properties. This model is based on aversion to open spaces, which is seen as the animal spending more time in the enclosed arms of the maze. This chapter describes the steps necessary for setting up and conducting the test, along with interpretation of the results.

The Forced Swim Test for Depression-Like Behavior in Rodents.

The forced swim test assesses learned helplessness, which is a feature of depression-like behavior in rodents. This test has also been used in testing the efficacy of existing and novel antidepressant drugs. It is based on the natural tendency of rodents to escape from water. Rodents are placed in a cylinder filled with water and the presumption is that those with a depression-like phenotype give up swimming earlier than those that are not depressed. Furthermore, antidepressant drugs reverse this effect. This chapter describes the basic setup and conduction of the test, along with interpretation of the results. It should be emphasized that this test should be conducted as part of a series of behavioral assessments in order to increase the accuracy of the results.

Object Burying Test for Assessment of Obsessive Compulsive Behaviors in Mice.

Obsessive-compulsive disorder (OCD) can occur in several psychiatric illnesses such as autism spectrum disorders (ASD) and it is more prevalent in children. This condition is characterized by repeated and apparently meaningless behaviors such as frequent hand washing, counting, tapping, and rocking. This can disrupt normal socialization and in some cases lead to self-harm. Therefore there is interest in developing more effective therapies for individuals suffering from these conditions. This chapter describes how to conduct the mouse marble burying test as a sensitive measure of compulsive behaviors.

The Nest Building Test in Mice for Assessment of General Well-Being.

This chapter presents a method for assessing general behavior, well-being, and sensorimotor gating. A detailed protocol is given for assessment of nest building performance in female mice using a strict scoring system. The test can be used for assessing moods and behaviors associated with psychiatric disorders such as depression and schizophrenia. It can also be useful for testing movement disorders such as Parkinson disease.

Free Dyadic Social Interaction Test in Mice.

Psychiatric disorders affect approximately one quarter of people worldwide at some point in their lifetime. This chapter provides a step-by-step guide to conduct behavioral tests in adult mice for investigations of social behavior, without the need for specific equipment. This test should allow the identification of key abnormalities in social interactions that can be followed up by targeted, more complex, behavioral analysis aimed at identification of new biomarkers and potential drug targets.

The Open Field Test for Measuring Locomotor Activity and Anxiety-Like Behavior.

The open field test is used in studies of the neurobiological basis of anxiety and screening for novel drug targets and anxiolytic compounds. This test uses a camera to measure movement of the test animal in the peripheral and central zones of a 42 × 42 × 42 cm polyvinyl chloride box. This chapter describes a protocol for carrying out the open-field test for assessment of locomotion and anxiety-like behavior in mice.

The Y-Maze for Assessment of Spatial Working and Reference Memory in Mice.

The Y-maze can be used to assess short term memory in mice. Spontaneous alternation, a measure of spatial working memory, can be assessed by allowing mice to explore all three arms of the maze and is driven by an innate curiosity of rodents to explore previously unvisited areas. A mouse with intact working memory, and hence intact prefrontal cortical functions, will remember the arms previously visited and show a tendency to enter a less recently visited arm. Spatial reference memory, which is underlined by the hippocampus, can also be tested by placing the test mice into the Y-maze with one arm closed off during training. After an inter-trial interval of for example 1 h, the mouse should remember which arm it has not explored previously and should visit this arm more often. This chapter describes the pre-test conditions, the materials required and the protocol for conducting and interpreting the results of these two related tests.