Aiginition Longitudinal Biomarker Investigation Of Neurodegeneration (ALBION): study design, cohort description, and preliminary data.

Objectives: Aiginition Longitudinal Biomarker Investigation Of Neurodegeneration (ALBION) is a longitudinal ongoing study initiated in 2018 that takes place in the Cognitive Disorders Clinic of Aiginition Hospital of the National and Kapodistrian University of Athens. Its aim is to address several research questions concerning the preclinical and prodromal stage of Alzheimer's disease and explore potential markers for early detection, prediction, and primary prevention of dementia. Methods: We here present the design and the preliminary baseline characteristics of ALBION. The sample of our study consists of people aged over 50 who are concerned about their memory but are cognitively normal (CN) or have mild cognitive deficits. Each participant undergoes an extensive assessment including several demographic, medical, social, environmental, clinical, nutritional, neuropsychological determinants and lifestyle activities. Furthermore, we are collecting data from portable devices, neuroimaging techniques and biological samples (blood, stools, CSF). All participants are assessed annually for a period of 10 years. Results: In total, 47 participants have completed the initial evaluation up to date and are divided in two groups, CN individuals (N = 26) and MCI patients (N = 21), based on their cognitive status. The participants are, on average, 64 years old, 46.3% of the sample is male with an average of 12.73 years of education. MCI patients report more comorbidities and have a lower score in the MMSE test. Regarding APOE status, 2 participants are ε4 homozygotes and 10 ε4 heterozygotes. CSF analyses (Aß42, Τ-tau, P-tau) revealed no differences between the two groups. Conclusion: The ALBION study offers an opportunity to explore preclinical dementia and identify new and tailored markers, particularly relating to lifestyle. Further investigation of these populations may provide a wider profile of the changes taking place in the preclinical phase of dementia, leading to potentially effective therapeutic and preventive strategies.

Incidence and predictors of post-stroke aphasia: the Arcadia Stroke Registry.

BACKGROUND AND PURPOSE: Aphasia is an important post-stroke sequela. We estimated the prevalence and main determinants of post-stroke aphasia in the prefecture of Arcadia, Greece. METHOD: Prospective population-based study of Arcadia residents diagnosed with first ever stroke. within a 2 year period. Associations of aphasia with potential predictors were analysed by logistic regression in: (i) the entire cohort and (ii) the sub-cohort of patients who were alive 28 days post-stroke. Multivariate regression models were adjusted for left hemispheric stroke localization and modified Rankin Scale 28 days post-stroke (MRS-1mo). RESULTS: Of 555 subjects, 126 (22.7%) suffered from aphasia. When only the 405 survivors were considered, 77 (19.0%) suffered from aphasia. In adjusted multivariate models of the entire cohort, factors conferring significantly higher risk for aphasia included female gender, diabetes mellitus (borderline significance) and heart disease. In adjusted multivariate models of survivors, only diabetes was associated with significantly higher risk for aphasia. CONCLUSIONS: Female gender, diabetes and heart disease were independent prospective predictors of aphasia. The study offers a quantitative estimate of the public health problem of post-stroke aphasia in Greece and suggests that the role of diabetes in post-stroke aphasia may be more important than previously appreciated.

Dietary lipids and geriatric depression scale score among elders: the EPIC-Greece cohort.

In a prospective epidemiological investigation aiming to identify dietary lipids potentially associated with affective state and depression, 610 healthy men and women aged 60 years or older, participating in the EPIC-Greece cohort and residing in the Attika region had dietary, sociodemographic, anthropometric, medical and lifestyle variables ascertained at enrollment. Six to 13 years later, affective state was evaluated through the 15-point geriatric depression scale (GDS) score along with cognitive function and medical variables. In multivariate linear regression analysis, while adjusting for potential confounders, GDS score was negatively associated with dietary intake of monounsaturated lipids (MUFA) and their main source, olive oil, and positively associated with intake of polyunsaturated lipids (PUFA) and one of their principal sources, seed oils. Intake of calories, total lipids, fish and seafood or saturated lipids did not exhibit significant association with GDS. Potential non-linearities were assessed by quantile multivariate regression analysis: The median GDS score was positively associated with PUFA and seed oils intake, while other lipid groups showed no appreciable associations. The 90th percentile of the GDS score (towards the high end) exhibited significant negative associations with MUFA and olive oil, weaker positive associations with PUFA and seed oils and no appreciable association with other lipid group dietary intakes. We conclude that among Attika elders, lower intake of seed oils and higher intake of olive oil prospectively predict a healthier affective state. Olive oil intake, in particular, predicts a lower chance of scoring in the highest part of the GDS.

Diet, physical activity and cognitive impairment among elders: the EPIC-Greece cohort (European Prospective Investigation into Cancer and Nutrition).

OBJECTIVES: To identify dietary and lifestyle variables that may affect cognitive function in the elderly. DESIGN: Population-based prospective cohort study. SETTING: General community residing in Athens and the surrounding Attica region of Greece. SUBJECTS: A total of 732 men and women, 60 years or older, participating in the EPIC-Greece cohort (European Prospective Investigation into Cancer and Nutrition) and residing in the Attica region had sociodemographic, anthropometric, medical, dietary and lifestyle variables ascertained at enrolment (1993-1999). Six to 13 years later, cognitive function was evaluated through the Mini-Mental State Examination (MMSE) score and affective state through the Geriatric Depression Scale (GDS). RESULTS: MMSE score was positively associated with years of formal education, height and physical activity and inversely with age, diabetes mellitus and GDS score (P < 0.05 for all). Among dietary variables, intake of PUFA was inversely associated with cognitive function and this association was largely accounted for by a similar association with seed oils. Adherence to the Mediterranean diet, as well as intake of olive oil, MUFA and SFA exhibited weakly positive but not significant associations. CONCLUSION: Physical activity and early life factors as reflected in height are significant predictors of cognitive function in the elderly. Seed oil consumption may adversely affect cognition, whereas other nutritional factors do not appear to have a quantitatively large effect.

Voltage-dependent calcium channels in ventromedial hypothalamic neurones of postnatal rats: modulation by oestradiol and phenylephrine.

Oestradiol actions in the hypothalamus play an important role in reproductive behaviour. Oestradiol treatment in vivo induces alpha(1b)-adrenoceptor mRNA and increases the density of alpha(1B)-adrenoceptor binding in the hypothalamus. Oestradiol is also known to modulate neuronal excitability, in some cases by modulating calcium channels. We assessed the effects of phenylephrine, an alpha(1)-adrenergic agonist, on low-voltage-activated (LVA) and high-voltage-activated (HVA) calcium channels in ventromedial hypothalamic (VMN) neurones from vehicle- and oestradiol-treated female rats. Whole-cell and gramicidin perforated-patch recordings were obtained, with barium as the charge carrier. In the absence of phenylephrine, oestradiol treatment increased the magnitude of LVA currents compared to controls, but had no effect on HVA currents. Phenylephrine enhanced HVA currents in a significantly greater proportion of neurones from oestradiol-treated rats (76%) than from vehicle-treated (41%) rats. The L-channel blocker nifedipine abolished this oestradiol effect on phenylephrine-enhanced HVA currents. Preincubating slices with the N-type channel blocker omega-conotoxin GVIA completely blocked the phenylephrine response, suggesting that the N-type channel is essential. Phenylephrine also stimulated LVA currents in approximately two-thirds of neurones in slices from both vehicle- and oestradiol-treated rats. Our data show that oestradiol increases LVA currents in the VMN. Oestradiol also amplifies alpha(1)-adrenergic signalling by increasing the proportion of neurones showing phenylephrine-stimulated HVA currents mediated by N- and L-type calcium channels. In this way, oestradiol may increase excitatory responses to arousing adrenergic inputs to VMN neurones governing oestradiol-dependent reproductive behaviour.

Rare association of multiple sclerosis and systemic lupus erythematosus.

Although both multiple sclerosis (MS) and systemic lupus erythematosus (SLE) are relatively common autoimmune disorders, especially in young women and often coexist in families, they are only rarely reported to coexist in a single patient. We here present a case of a young woman with a history of MS from many years who diagnosed as suffering as well from SLE.

Ca(2+)-dependent inactivation of NMDA receptors: fast kinetics and high Ca2+ sensitivity in rat dorsal horn neurons.

1. Ca(2+)-dependent inactivation (CDI) of NMDA receptors was studied using both cultured embryonic rat dorsal horn neurons and acutely dissociated postnatal rat dorsal horn neurons. The perforated patch recording method was employed in order to preserve intracellular Ca2+ buffers and other cellular constituents. In this way, the kinetics of intracellular Ca2+ concentration ([Ca2+]i) transients and other second messenger signalling systems were maintained in a relatively normal condition. 2. Continuous application of 30 microM NMDA to cultured dorsal horn neurons voltage clamped at -70 mV evoked currents that inactivated to about 40% of the peak value with time constants between 200 and 600 ms. CDI with similar kinetics was also observed in acutely dissociated postnatal rat dorsal horn neurons. 3. When NMDA was applied in a low (20 microM) Ca2+ bath or when dorsal horn neurons were held at +70 mV, inactivation was either very weak or absent. The peaks of NMDA currents were significantly suppressed when preceded by voltage steps to 0 mV or by evoked action potentials. The suppression was dependent on the presence of Ca2+ in the extracellular solution. Voltage steps to +100 mV were ineffective in suppressing NMDA responses. Therefore, the observed inactivation was caused by an increase in [Ca2+]i following Ca2+ entry through NMDA channels or through voltage gated Ca2+ channels. 4. Caffeine application reduced currents evoked by subsequent NMDA applications. This reduction was not dependent on the presence of extracellular Ca2+ but was abolished after incubation of the cells with ryanodine, suggesting that Ca2+ release from intracellular stores also induced CDI. 5. Simultaneous measurements of somal [Ca2+]i and of currents evoked by somal NMDA applications showed that the magnitude of CDI was correlated with [Ca2+]i levels and that [Ca2+]i elevations of 100-300 nM were usually sufficient to inactivate NMDA currents by more than 30%. 6. Dose-response curves of non inactivated and inactivated NMDA responses showed that the apparent receptor affinity for NMDA is not different under the two conditions. CDI is caused instead by non-competitive inhibition of NMDA receptors. CDI was not overcome by increasing glycine concentration, suggesting that it is not mediated by glycine dissociation from the receptor. 7. These results show that, with an intact intracellular environment, CDI in dorsal horn neurons constitutes a potent, inhibitory control of NMDA currents with a faster onset than previously demonstrated. CDI is induced by a variety of [Ca2+]i-elevating stimuli of physiological relevance including Ca2+ entry through ligand- and voltage-gated channels and Ca2+ release from intracellular stores. Our demonstration that CDI is strongly expressed in neurons maturing in vivo supports the hypothesis that CDI may regulate, in part, the postsynaptic integration of excitatory input in the mature or maturing nervous system.

Calcium entry through a subpopulation of AMPA receptors desensitized neighbouring NMDA receptors in rat dorsal horn neurons.

1. A Ca(2+)-dependent interaction between non-NMDA and NMDA receptors was studied in embryonic rat dorsal horn neurons grown in tissue culture using perforated-patch recording. Specifically, non-NMDA receptors were found to induce reversible inhibition of NMDA receptors in a manner dependent on the presence of extracellular Ca2+. 2. Non-NMDA receptor-induced inhibition of NMDA receptors was mediated by the elevation of intracellular Ca2+ concentration produced by Ca2+ entry through a subpopulation of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) non-NMDA receptors. Furthermore, Ca2+ entry through the AMPA channels alone is sufficient for desensitization of NMDA channels to occur. 3. Imaging of neuritic sites of Ca2+ revealed that Ca(2+)-permeable AMPA channels are often co-localized with NMDA channels on dorsal horn neurons, indicating that the Ca(2+)-mediated interaction between receptors may occur within small dendritic domains. 4. The ability of Ca(2+)-permeable AMPA channels to inhibit adjacent NMDA channels may contribute to the postsynaptic integration of excitatory input.

Perforated-patch recording with gramicidin avoids artifactual changes in intracellular chloride concentration.

The antibiotic gramicidin, when incorporated into lipid membranes, forms pores that are exclusively permeable to monovalent cations and small unchanged molecules. We report the use of gramicidin for perforated patch-clamp recordings in the whole-cell mode. Recordings were performed in cultured rat spinal cord dorsal horn neurons. Cells had stable resting potentials and series resistances for times routinely exceeding 60 min. To test if intracellular chloride concentration ([Cl]i) remains stable with this technique, we measured responses to agonists of glycine and GABAA receptors, both of which gate chloride conductances. The driving force for these responses remained stable at values that differed significantly from values that would be expected if [Cl-]i were biased towards pipette [Cl-]. We conclude that gramicidin perforated-patch recording, in addition to other properties of the perforated-patch recording technique, has the advantage of not altering [Cl-]i. It is, therefore, an electrophysiological method particularly suitable for studies of anionic channels when [Cl-]i is a variable of interest, as well as for studies of homeostatic [Cl-]i regulation.

Developmental loss of GABA- and glycine-induced depolarization and Ca2+ transients in embryonic rat dorsal horn neurons in culture.

More than 90% of dorsal horn neurons from embryonic day 15-16 rats responded to the inhibitory amino acids GABA and glycine by a transient elevation of intracellular Ca2+ concentration ([Ca2+]i) when maintained in culture for < 1 week. This [Ca2+]i response has previously been shown to be due to depolarization and subsequent Ca2+ entry through voltage-gated Ca2+ channels following activation of bicuculline-sensitive GABAA receptors and strychnine-sensitive glycine receptors. Both the number of cells responding to GABA and glycine and the amplitude of the [Ca2+]i response diminished over time in culture. By 30 days in culture, none of the cells responded to GABA, muscimol or glycine by elevation of [Ca2+]i. The loss of the [Ca2+]i response was not due to a change in the abundance or the properties of voltage-gated Ca2+ channels, since over the same period of time dorsal horn neurons showed a large increase in the amplitude of the [Ca2+]i transient in response to 30 mM K+. Nor was the loss of the [Ca2+]i response due to a loss of GABA and glycine receptors. Instead, the decrease in the [Ca2+]i response over time paralleled a similar change in the electrophysiological responses. More than 90% of the neurons tested were depolarized in response to inhibitory amino acids during the first week in culture. After 30 days, all neurons tested responded to GABA and glycine with a hyperpolarization. These observations add support to the suggestion that GABA and glycine may excite dorsal horn neurons early in development and play a role in postmitotic differentiation.

Mechanisms of GABA and glycine depolarization-induced calcium transients in rat dorsal horn neurons.

1. The mechanisms and effects of GABA- and glycine-evoked depolarization were studied in cultured rat dorsal horn neurons using indo-1 recordings of [Ca2+]i and patch clamp recordings in conventional whole-cell or perforated-patch mode. 2. Application of GABA to unclamped neurons caused [Ca2+]i increases that were dose dependent and exhibited GABAA receptor pharmacology. Calcium entered the neurons via high-threshold voltage-gated calcium channels (conotoxin and nimodipine sensitive). 3. In perforated-patch recordings employing cation-selective ionophores, GABAA receptor activation depolarized 123 of 132 cells to membrane potentials as depolarized as -33 mV (mean -50 mV in all 132 cells, +12 mV above resting potential). The ionic basis of the depolarization was determined by extracellular ion substitution; increased anionic conductance could account fully for the results. 4. Glycine, acting at a strychnine-sensitive receptor, also caused Ca2+ entry into these neurons through voltage-gated Ca2+ channels. Glycine and GABA both evoked [Ca2+]i responses in the same cells and the responses were highly correlated in amplitude. Glycine also depolarized all five cells tested with perforated recording. Each of the five cells was also depolarized by muscimol to a value similar to that obtained for glycine. 5. Both the depolarization and the increases in [Ca2+]i caused by GABA and glycine could potentially play a role in processes of development and differentiation and sensory transmission in the spinal cord dorsal horn.