Edna N. Yamasaki

GABAergic system in the developing mammalian retina: dual sources of GABA at early stages of postnatal development

In the present work, we have characterized the maturation of the GABAergic system in mammalian retina. Immunoreactivity for GABA, GAD (glutamic acid decarboxylase, EC 4.1.1.15) ‐65 and ‐67 in the adult rat retina was localized in cells in the inner nuclear and ganglion cell layers. This pattern was established around postnatal day 8 and included transient GABA and GAD‐67 expression in horizontal cells. GAD activity was very low at P1 and P4, increasing after P8, reaching maximal activity by P21 and decreasing to attain adult values by P30. GABA content was approximately constant from P1 to P13, increasing thereafter to reach adult levels. GAD protein content increased progressively with postnatal development and the two isoforms could be distinguished at P8.

Nicotinic acetylcholine receptor activation reduces skeletal muscle inflammation of mdx mice.

Mdx mice develop an inflammatory myopathy characterized at different ages by myonecrosis with scattered inflammatory infiltrates followed by muscular regeneration and later persistent fibrosis. This work aimed to verify the putative anti-inflammatory role of nicotinic acetylcholine receptor (nAChR) in the mdx muscular lesion. Mitigation of myonecrosis and decreased TNFα production were accompanied by increased numbers of F4/80 macrophages expressing nAChRα7. In vivo treatment with nicotine attenuated muscular inflammation characterized by reduced metalloprotease MMP-9 activity, TNFα and NFkB content and increased muscular regeneration. Our data indicate that nAChR activation influences local inflammatory responses in the muscular lesion of mdx mice.

Cationic homopolymer model networks and star polymers: synthesis by group transfer polymerization and characterization of the aqueous degree of swelling

Abstract A series of hydrophilic model networks based on homopolymers of a weak base were prepared using group transfer polymerization (GTP) and characterized in terms of their aqueous degree of swelling as a function of pH. This constitutes the first reported example of cationic hydrogels in which the length of segments between cross-links is kept constant. The synthesis involved the GTP of 2-(dimethylamino)ethyl methacrylate (DMAEMA, weak base monomer) in tetrahydrofuran (THF, solvent) using the bifunctional initiator, 1,4-bis(methoxytrimethylsiloxymethylene)-cyclohexane (MTSMC) and the subsequent in situ cross-linking with the addition of 8-fold mole excess with respect to the initiator of the difunctional monomer ethyleneglycol dimethacrylate (EGDMA). Four different model networks with degrees of polymerization (DP) of the linear segments between cross-links of 10, 20, 50 and 100 were prepared. The molecular weight (MW) and molecular weight distribution (MWD) of the linear segments were measured using gel permeation chromatography (GPC) in THF. The degree of swelling of all model networks was measured in water as a function of the solution pH. The conditions of gel synthesis were optimized with respect to cross-linker loading and monomer concentration using the results of a parallel study on the synthesis of star polymers of DMAEMA. The star polymers were also prepared by GTP of DMAEMA and linking (to stars) with EGDMA, but a monofunctional initiator, 1-methoxy-1-trimethylsiloxy-1-methyl-propene (MTS), was used rather than MTSMC. The star polymers were analyzed by GPC which revealed the percentage of free homopolymer relative to star polymer. The optimal conditions were identified as those under which the percentage of unlinked (free) homopolymer is minimized — the same conditions were adopted for the synthesis of the networks.

Extreme weather and air pollution effects on cardiovascular and respiratory hospital admissions in Cyprus.

In many regions of the world, climatic change is associated with increased extreme temperatures, which can have severe effects on mortality and morbidity. In this study, we examine the effect of extreme weather on hospital admissions in Cyprus, for inland and coastal areas, through the use of synoptic weather classifications (air mass types). In addition, the effect of particulate air pollution (PM10) on morbidity is examined. Our results show that two air mass types, namely (a) warm, rainy days with increased levels of water vapour in the atmosphere and (b) cold, cloudy days with increased levels of precipitation, were associated with increased morbidity in the form of hospital admissions. This was true both for cardiovascular and respiratory conditions, for all age groups, but particularly for the elderly, aged over 65. Particulate air pollution was also associated with increased morbidity in Cyprus, where the effect was more pronounced for cardiovascular diseases.

Norepinephrine acts as D1-dopaminergic agonist in the embryonic avian retina: Late expression of β1-adrenergic receptor shifts norepinephrine specificity in the adult tissue

Dopamine is the main catecholamine found in the chick retina whereas norepinephrine is only found in trace amounts. We compared the effectiveness of dopamine and norepinephrine in promoting cyclic AMP accumulation in retinas at embryonic day 13 (E13) and from post-hatched chicken (P15). Dopamine (EC(50)=10microM) and norepinephrine (EC(50)=30microM), but not the beta(1)-adrenergic agonist isoproterenol, stimulated over seven-fold the production of cyclic AMP in E13 retina. The cyclic AMP accumulation induced by both catecholamines in embryonic tissue was entirely blocked by 2microM SCH23390, a D(1) receptor antagonist, but not by alprenolol (beta-adrenoceptor antagonist). In P15 retinas, 100microM isoproterenol stimulated five-fold the accumulation of cAMP. This effect was blocked by propanolol (10microM), but not by 2microM SCH23390. Embryonic and adult retina display beta(1) adrenergic receptor mRNA as detected by RT-PCR, but the beta(1) adrenergic receptor protein was detected only in post-hatched tissue. We conclude that norepinephrine cross-reacts with D(1) dopaminergic receptor with affinity similar to that of dopamine in the embryonic retina. In the mature retina, however, D(1) receptors become restricted to activation by dopamine. Moreover, as opposed to the embryonic tissue, norepinephrine seems to stimulate cAMP accumulation via beta(1)-like adrenergic receptors in the mature tissue.

Opposite roles of GABA and excitatory amino acids on the control of GAD expression in cultured retina cells

The mechanism of control of GAD expression by GABA and excitatory amino acids (EAAs) was studied in chick and rat retina cultures using immunohistochemical and PAGE-immunoblot detection of the enzyme, as well as by measuring enzyme activity. Aggregate cultures were prepared with retina cells obtained from chick embryos at embryonic days 8-9 (E8-E9). Organotypical cultures were also prepared with retinas from E14 chick embryos, post-hatched chicken and P21 rats. GABA (1-20 mM) fully prevented GAD expression in aggregate and organotypical cultures from chick embryo retinas. A substantial, but not complete, reduction of GAD was also observed in organotypical cultures of post-hatched chicken and P21 rats, in which both forms of the enzyme (GAD65 and 67) were affected. The GABA effect was not mimicked by THIP (100 microM), baclofen (100 microM) or CACA (300 microM), agonists of GABAa, b and c receptors, respectively. NNC-711, a potent inhibitor of GABA transporters, reduced by 50% the inhibition of GAD activity promoted by GABA. Aggregates exposed to GABA and treated with glutamate (5 mM) or kainate (100 microM) displayed an intense GAD-like immunoreactivity in many cell bodies, but not in neurite regions. Immunoblot analysis revealed that the increase in GAD-like immunoreactivity by EAA corresponded to a 67-kDa protein. However, GAD activity was not detected. Treatment of aggregates or retina homogenates with SNAP, a NO producing agent (but not its oxidized form), reduced GAD activity by more than 60% indicating that the lack of enzyme activity in GAD-like immunoreactive cells, could be due to NO production by EAA stimulation.

Heat-related mortality in Cyprus for current and future climate scenarios

Extreme temperatures have long been associated with adverse health impacts, ranging from minor illness, to increased hospitalizations and mortality. Heat-related mortality during summer months is likely to become an increasing public health problem in future due to the effects of climate change. We performed a health impact assessment for heat-related mortality for the warm months of April-September for the years 2004 to 2009 inclusive, for the city of Nicosia and for Cyprus as a whole, based on separately derived exposure-response functions. We further estimated the potential future heat-related mortality by including climate projections for southern Europe, which suggest changes in temperature of between 1°C and 5°C over the next century. There were 32 heat-related deaths per year in Cyprus over the study period. When adding the projected increase in temperature due to climate change, there was a substantial increase in mortality: for a 1°C increase in temperature, heat related mortality in Cyprus was estimated to double to 64 per year, and for a 5°C increase, heat-related mortality was expected to be 8 times the baseline rate for the warm season (281 compared with 32). This analysis highlights the importance of preparing for potential health impacts due to heat in Cyprus, particularly under a changing climate.

Human mortality in Cyprus: the role of temperature and particulate air pollution

AbstractnClimatic change results in increased occurrence of heat waves, and the thermal stress caused by such phenomena is leading to higher levels of heat-related mortality worldwide. This study is the first to examine the effect of extreme weather on mortality in Cyprus. It investigates the individual effect of meteorological indicators on mortality, as well as the role of particulate air pollution (PM10). A generalized linear model (GLM) with quasi-Poisson regression was implemented. GLM included a temperature function and was adjusted for relative humidity and seasonality. The temperature function was developed under a newly developed framework of distributed lag nonlinear models, which capture nonlinearities and delayed effects of heat simultaneously. GLM was extended to examine the confounding effect of air pollution. All the results on heat effects are presented. High temperatures had a significant effect on mortality with increased mortality rates, independent of humidity and seasonality. Mortality risk increased steeply above a temperature threshold. A direct heat effect was shown, with higher risk on the current and next day of a severe heat event. PM10 was not found to have a confounding effect on the temperature–mortality relationship, since the strength of this relationship remained after the inclusion of PM10 in the model. Differences existed between urban and coastal areas.

Ethanol increases GABA release in the embryonic avian retina

Several mechanisms underlying ethanol action in GABAergic synapses have been proposed, one of these mechanisms is on GABA release. Here, we report that in ovo exposure to ethanol induces an increase on GABA release in the embryonic chick retina. Eleven‐day‐old chick embryos (E11) received an injection of either phosphate buffer saline (PBS) or ethanol (10%, v/v, diluted in PBS), and were allowed to develop until E16. A single glutamate stimulus (2 mM) showed approximately a 40% increase on GABA release in E16 retinas when compared to controls. The effect was dependent on NMDA receptors and GAD65 mRNA levels, which were increased following the ethanol treatment. However, the numbers of GABA‐, GAD‐, and NR1‐immunoreactive cells, and the expression levels of these proteins, were not affected. We conclude that ethanol treatment at a time point when synapses are being formed during development selectively increases GABA release in the retina via a NMDA receptor‐dependent process.

Impact of ethanol on the developing GABAergic system.

Alcohol intake during pregnancy has a tremendous impact on the developing brain. Embryonic and early postnatal alcohol exposures have been investigated experimentally to elucidate the fetal alcohol spectrum disorders (FASD) milieu, and new data have emerged to support a devastating effect on the GABAergic system in the adult and developing nervous system. GABA is a predominantly inhibitory neurotransmitter that during development excites neurons and orchestrates several developmental processes such as proliferation, migration, differentiation, and synaptogenesis. This review summarizes and brings new data on neurodevelopmental aspects of the GABAergic system with FASD in experimental telencephalic models. Anat Rec, 292:1922–1939, 2009.