Aberrant oscillatory activity in neurofibromatosis type 1: an EEG study of resting state and working memory.

BACKGROUND: Neurofibromatosis type 1 (NF1) is a genetic neurodevelopmental disorder commonly associated with impaired cognitive function. Despite the well-explored functional roles of neural oscillations in neurotypical populations, only a limited number of studies have investigated oscillatory activity in the NF1 population. METHODS: We compared oscillatory spectral power and theta phase coherence in a paediatric sample with NF1 (N = 16; mean age: 13.03 years; female: n = 7) to an age/sex-matched typically developing control group (N = 16; mean age: 13.34 years; female: n = 7) using electroencephalography measured during rest and during working memory task performance. RESULTS: Relative to typically developing children, the NF1 group displayed higher resting state slow wave power and a lower peak alpha frequency. Moreover, higher theta power and frontoparietal theta phase coherence were observed in the NF1 group during working memory task performance, but these differences disappeared when controlling for baseline (resting state) activity. CONCLUSIONS: Overall, results suggest that NF1 is characterised by aberrant resting state oscillatory activity that may contribute towards the cognitive impairments experienced in this population. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03310996 (first posted: October 16, 2017).

Are mothers less likely to breastfeed in harsh environments? Physical environmental quality and breastfeeding in the Born in Bradford study.

We use the United Kingdom's Born in Bradford study to investigate whether women in lower quality environments are less likely to breastfeed. We use measures of physical environmental quality (water disinfectant by-products [DBPs], air pollution, passive cigarette smoke, and household condition) alongside socio-economic indicators, to explore in detail how different exposures influence breastfeeding. Drawing on evolutionary life history theory, we predict that lower environmental quality will be associated with lower odds of initiating, and higher hazards of stopping, breastfeeding. As low physical environmental quality may increase the risk of adverse birth outcomes, which may in turn affect breastfeeding chances, we also test for mediation by gestational age, birthweight, head circumference, and abdominal circumference. Our sample is composed of mothers who gave birth at the Bradford Royal Infirmary in West Yorkshire between March 2007 and December 2010 for whom breastfeeding initiation data was available. Analyses were stratified by the two largest ethnic groups: White British (n = 3,951) and Pakistani-origin (n = 4,411) mothers. After controlling for socio-economic position, Pakistani-origin mothers had lower chances of initiating and higher chances of stopping breastfeeding with increased water DBP exposure (e.g., OR for 0.03-0.61 vs. <0.02 µg/day dibromochloromethane exposure 0.70 [0.58-0.83], HR 1.16 [0.99-1.36]), greater air pollution exposure predicted lower chances of initiation for both ethnic groups (e.g., OR for 10 µg/m3 increase in nitrogen dioxide 0.81 [0.66-0.99] for White British mothers and 0.79 [0.67-0.94] for Pakistani-origin mothers) but also a reduced hazard of stopping breastfeeding for White British mothers (HR 0.65 [0.52-0.80]), and exposure to household damp/mould predicted higher chances of breastfeeding initiation amongst White British mothers (OR 1.66 [1.11-2.47]). We found no evidence that physical environmental quality effects on breastfeeding were mediated through birth outcomes amongst Pakistani-origin mothers and only weak evidence (p < 0.10) amongst White British mothers (exposure to passive cigarette smoke was associated with having lower birthweight infants who were in turn less likely to be breastfed whereas greater air pollution exposure was associated with longer gestations and in turn reduced hazards of stopping breastfeeding). Overall, our findings suggest that there is differential susceptibility to environmental exposures according to ethnicity. Although the water DBP results for Pakistani-origin mothers and air pollution-initiation results for both ethnic groups support our hypothesis that mothers exhibit reduced breastfeeding in poorer quality environments, several physical environmental quality indicators showed null or positive associations with breastfeeding outcomes. We consider physiological explanations for our findings and their implications for life history theory and public health policy.

Knockdown of both mitochondrial isocitrate dehydrogenase enzymes in pancreatic beta cells inhibits insulin secretion.

BACKGROUND: There are three isocitrate dehydrogenases (IDHs) in the pancreatic insulin cell; IDH1 (cytosolic) and IDH2 (mitochondrial) use NADP(H). IDH3 is mitochondrial, uses NAD(H) and was believed to be the IDH that supports the citric acid cycle. METHODS: With shRNAs targeting mRNAs for these enzymes we generated cell lines from INS-1 832/13 cells with severe (80%-90%) knockdown of the mitochondrial IDHs separately and together in the same cell line. RESULTS: With knockdown of both mitochondrial IDH's mRNA, enzyme activity and protein level, (but not with knockdown of only one mitochondrial IDH) glucose- and BCH (an allosteric activator of glutamate dehydrogenase)-plus-glutamine-stimulated insulin release were inhibited. Cellular levels of citrate, α-ketoglutarate, malate and ATP were altered in patterns consistent with blockage at the mitochondrial IDH reactions. We were able to generate only 50% knockdown of Idh1 mRNA in multiple cell lines (without inhibition of insulin release) possibly because greater knockdown of IDH1 was not compatible with cell line survival. CONCLUSIONS: The mitochondrial IDHs are redundant for insulin secretion. When both enzymes are severely knocked down, their low activities (possibly assisted by transport of IDH products and other metabolic intermediates from the cytosol into mitochondria) are sufficient for cell growth, but inadequate for insulin secretion when the requirement for intermediates is certainly more rapid. The results also indicate that IDH2 can support the citric acid cycle. GENERAL SIGNIFICANCE: As almost all mammalian cells possess substantial amounts of all three IDH enzymes, the biological principles suggested by these results are probably extrapolatable to many tissues.

Assessing mood in older adults: a conceptual review of methods and approaches.

BACKGROUND: Accurate measures of mood state are important for understanding and optimizing health and well-being in later life. A range of different mood assessment measures is available, reflecting the variety of ways in which mood has been conceptualized and the different purposes for which measures have been developed. METHODS: We undertook a conceptual review of the literature relating to mood and its assessment in older populations. RESULTS: Moods are subjective states of mind that are typically described and quantified using self-report measures. Moods can be conceptually differentiated from the related psychological concepts of emotion, well-being, quality of life, and depression. Quantitative tools for assessing mood state include single-item mood ratings, composite factor scales, and clinical depression assessments. Mood assessments may be administered retrospectively or contemporaneously to the mood state of interest. The method and temporal perspective used to assess mood state will impact on the nature and precision of the mood data that are collected, and the types of research questions that can be addressed. CONCLUSIONS: No single mood assessment technique can be considered optimal for all situations. Rather, both the type of tool and the temporal perspective taken must be selected according to the nature of the study design and the research question being addressed. More thorough and frank reporting of the rationale for, and limitations of, mood assessment techniques are also essential for continued development of mood research with older adults.

Differences between human and rodent pancreatic islets: low pyruvate carboxylase, atp citrate lyase, and pyruvate carboxylation and high glucose-stimulated acetoacetate in human pancreatic islets.

Anaplerosis, the net synthesis in mitochondria of citric acid cycle intermediates, and cataplerosis, their export to the cytosol, have been shown to be important for insulin secretion in rodent beta cells. However, human islets may be different. We observed that the enzyme activity, protein level, and relative mRNA level of the key anaplerotic enzyme pyruvate carboxylase (PC) were 80-90% lower in human pancreatic islets compared with islets of rats and mice and the rat insulinoma cell line INS-1 832/13. Activity and protein of ATP citrate lyase, which uses anaplerotic products in the cytosol, were 60-75% lower in human islets than in rodent islets or the cell line. In line with the lower PC, the percentage of glucose-derived pyruvate that entered mitochondrial metabolism via carboxylation in human islets was only 20-30% that in rat islets. This suggests human islets depend less on pyruvate carboxylation than rodent models that were used to establish the role of PC in insulin secretion. Human islets possessed high levels of succinyl-CoA:3-ketoacid-CoA transferase, an enzyme that forms acetoacetate in the mitochondria, and acetoacetyl-CoA synthetase, which uses acetoacetate to form acyl-CoAs in the cytosol. Glucose-stimulated human islets released insulin similarly to rat islets but formed much more acetoacetate. ß-Hydroxybutyrate augmented insulin secretion in human islets. This information supports previous data that indicate beta cells can use a pathway involving succinyl-CoA:3-ketoacid-CoA transferase and acetoacetyl-CoA synthetase to synthesize and use acetoacetate and suggests human islets may use this pathway more than PC and citrate to form cytosolic acyl-CoAs.

Differential effects of delirium on fluid and crystallized cognitive abilities.

Patients with delirium (acute confusional state) show extensive cognitive deficits. These deficits have typically been measured using tests of fluid cognition, which involve the active processing of mental representations. However, the effects of delirium on stored, crystallized dimensions of cognition, such as well-learnt word pronunciation knowledge, are not known. In this study 37 patients (aged 60-85 years) without delirium were recruited before undergoing cardiac surgery. Cognitive assessments were performed 0-8 days before surgery and again 2-9 days after surgery in order to determine the effects of post-operative delirium (POD) on fluid and crystallized aspects of cognition. Crystallized cognition was tested with the National Adult Reading Test (NART). Fluid cognition was tested with digit span, verbal fluency and Stroop tests. Nine patients (24%) developed delirium post-operatively. Patients with delirium showed significant post-operative deficits on most tests of fluid cognition, but no change in the NART measure of crystallized cognition (p=0.95). These results parallel recent findings in Alzheimer's dementia and suggest that, despite showing extensive deficits of fluid cognitive processing, crystallized cognition is preserved in delirium. The results also suggest that the NART may be a useful tool for assessing premorbid ability in patients with delirium.

Use of cells expressing gamma subunit variants to identify diverse mechanisms of AMPK activation.

A wide variety of agents activate AMPK, but in many cases the mechanisms remain unclear. We generated isogenic cell lines stably expressing AMPK complexes containing AMP-sensitive (wild-type, WT) or AMP-insensitive (R531G) gamma2 variants. Mitochondrial poisons such as oligomycin and dinitrophenol only activated AMPK in WT cells, as did AICAR, 2-deoxyglucose, hydrogen peroxide, metformin, phenformin, galegine, troglitazone, phenobarbital, resveratrol, and berberine. Excluding AICAR, all of these also inhibited cellular energy metabolism, shown by increases in ADP:ATP ratio and/or by decreases in cellular oxygen uptake measured using an extracellular flux analyzer. By contrast, A769662, the Ca(2+) ionophore, A23187, osmotic stress, and quercetin activated both variants to varying extents. A23187 and osmotic stress also increased cytoplasmic Ca(2+), and their effects were inhibited by STO609, a CaMKK inhibitor. Our approaches distinguish at least six different mechanisms for AMPK activation and confirm that the widely used antidiabetic drug metformin activates AMPK by inhibiting mitochondrial respiration.

Chronic reduction of the cytosolic or mitochondrial NAD(P)-malic enzyme does not affect insulin secretion in a rat insulinoma cell line.

The cytosolic malic enzyme (ME1) has been suggested to augment insulin secretion via the malate-pyruvate and/or citrate-pyruvate shuttles, through the production of NADPH or other metabolites. We used selectable vectors expressing short hairpin RNA (shRNA) to stably decrease Me1 mRNA levels by 80-86% and ME1 enzyme activity by 78-86% with either of two shRNAs in the INS-1 832/13 insulinoma cell line. Contrary to published short term ME1 knockdown experiments, our long term targeted cells showed normal insulin secretion in response to glucose or to glutamine plus 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid. We found no increase in the mRNAs and enzyme activities of the cytosolic isocitrate dehydrogenase or glucose-6-phosphate dehydrogenase, which also produce cytosolic NADPH. There was no compensatory induction of the mRNAs for the mitochondrial malic enzymes Me2 or Me3. Interferon pathway genes induced in preliminary small interfering RNA experiments were not induced in the long term shRNA experiments. We repeated our study with an improved vector containing Tol2 transposition sequences to produce a higher rate of stable transferents and shortened time to testing, but this did not alter the results. We similarly used stably expressed shRNA to reduce mitochondrial NAD(P)-malic enzyme (Me2) mRNA by up to 95%, with severely decreased ME2 protein and a 90% decrease in enzyme activity. Insulin release to glucose or glutamine plus 2-aminobicyclo[2,2,1]heptane-2-carboxylic acid remained normal. The maintenance of robust insulin secretion after lowering expression of either one of these malic enzymes is consistent with the redundancy of pathways of pyruvate cycling and/or cytosolic NADPH production in insulinoma cells.

Acetoacetate and beta-hydroxybutyrate in combination with other metabolites release insulin from INS-1 cells and provide clues about pathways in insulin secretion.

Mitochondrial anaplerosis is important for insulin secretion, but only some of the products of anaplerosis are known. We discovered novel effects of mitochondrial metabolites on insulin release in INS-1 832/13 cells that suggested pathways to some of these products. Acetoacetate, beta-hydroxybutyrate, alpha-ketoisocaproate (KIC), and monomethyl succinate (MMS) alone did not stimulate insulin release. Lactate released very little insulin. When acetoacetate, beta-hydroxybutyrate, or KIC were combined with MMS, or either ketone body was combined with lactate, insulin release was stimulated 10-fold to 20-fold the controls (almost as much as with glucose). Pyruvate was a potent stimulus of insulin release. In rat pancreatic islets, beta-hydroxybutyrate potentiated MMS- and glucose-induced insulin release. The pathways of their metabolism suggest that, in addition to producing ATP, the ketone bodies and KIC supply the acetate component and MMS supplies the oxaloacetate component of citrate. In line with this, citrate was increased by beta-hydroxybutyrate plus MMS in INS-1 cells and by beta-hydroxybutyrate plus succinate in mitochondria. The two ketone bodies and KIC can also be metabolized to acetoacetyl-CoA and acetyl-CoA, which are precursors of other short-chain acyl-CoAs (SC-CoAs). Measurements of SC-CoAs by LC-MS/MS in INS-1 cells confirmed that KIC, beta-hydroxybutyrate, glucose, and pyruvate increased the levels of acetyl-CoA, acetoacetyl-CoA, succinyl-CoA, hydroxymethylglutaryl-CoA, and malonyl-CoA. MMS increased incorporation of (14)C from beta-hydroxybutyrate into citrate, acid-precipitable material, and lipids, suggesting that the two molecules complement one another to increase anaplerosis. The results suggest that, besides citrate, some of the products of anaplerosis are SC-CoAs, which may be precursors of molecules involved in insulin secretion.