A quantitative comparison of synaptic density following perfusion versus immersion fixation in the rat cerebral cortex

The main objective of the present study was to perform an unbiased comparison of immersion vs. perfusion techniques to assess whether we could use the former to quantify synapses through electron microscopy (EM). Using the immersion technique is ideally suited for instances in which the specimen under study could not be perfused under the standard EM protocol. Our results suggest that, despite suboptimal qualitative results, fixation by immersion allows for adequate quantification of synapses.     

Interaction of nitric oxide and endothelin-1 in ischemia/reperfusion injury of rat heart

Although several studies have demonstrated that nitric oxide appears to be cardioprotective and endothelin-1 (ET-1) deleterious in myocardial ischemia/reperfusion injury, their interactions in the intact heart are unknown. Therefore, coronary effluent and interstitial fluid ("transudate") levels of ET-1 and cyclic GMP, an indirect measure of nitric oxide production, were determined simultaneously in normoxic and reperfused hearts and compared with myocardial and coronary function. Rat hearts were buffer-perfused at 9 ml/min/g heart wet weight for 45 min (baseline), followed either by another 45 min of perfusion (normoxia), or 15 min of total global ischemia and 30 min reperfusion. Hearts received, from 42-90 min, either vehicle, the inhibitor of nitric oxide formation NG-nitro-L-arginine (L-NNA; 200 micromol/l), the nitric oxide donor S-nitroso-N-acetyl-DL-penicillamine (SNAP; 200 micromol/l), or the ET receptor antagonist PD 142893 (200 nmol/l). Both mediators were released preferentially into the vascular lumen which resulted in similar luminal and interstitial concentrations of cyclic GMP, but three-fold higher levels of ET-1 in tissue because of the higher effluent than transudate flow rate. L-NNA increased the release of ET-1 and worsened coronary function, whereas SNAP had opposite effects. On reperfusion, considerable functional impairment was observed, although levels of cyclic GMP both in the vascular and tissue compartment were not reduced, but even increased. Reperfusion functional impairment was aggravated after inhibiting the synthesis of nitric oxide, whereas SNAP restored cardiac and coronary function close to pre-ischemic level. Deterioration of function corresponded with an increased level, and improvement with a decreased level of intersitial ET-1 at the onset of reperfusion. PD 142893 was similarly cardioprotective as SNAP both in normoxia and reperfusion. These results suggest that in reperfusion, cardiac function is depressed, despite increased rather than decreased endogenous nitric oxide production, largely due to the prevalence of the deleterious effects of ET-1 which are overcome by antagonism of ET receptors or exogenous nitric oxide supplied by SNAP.     

Quantifying the influence of cold water intrusions in a shallow,coastal system across contrasting years: Green Bay,Lake Michigan

We present water column thermal structure for two climatically different years: 2012, which experienced abnormally warm spring and summer air temperatures preceded by a relatively low ice winter and 2013, which experienced cooler than average spring and average summer air temperatures and preceded by average ice conditions. Mean bottom water temperatures for the season and during cold water intrusions were significantly warmer in 2012 than 2013 leading to a significantly reduced stratified season in 2012. Cold water intrusions were driven into southern Green Bay by southerly winds while intrusions were terminated when winds switched to persistent northerly winds. 2012 observed a significant increase in northerly winds relative to 2013, decreasing cold water intrusion presence and duration but winds did not fully explain the difference in thermal conditions for southern Green Bay. These cold bottom waters drive stratification in polymictic southern Green Bay while dimictic waters were found to have significantly warmer bottom temperatures during 2012 and a deeper mixed layer. Our observations suggest that relatively shallow (<20?m), seasonally stratified systems may not increase in stratification strength and duration under a warming climate; rather, changing wind climatology and surface heat flux can inform the degree to which the mixing regime can be expected to change and impact stratification and thermal structure of coastal systems. We discuss the biogeochemical implications of different thermal regimes, particularly within the context of multiple drivers of physical water column structure in eutrophic, stratified coastal systems.     

Evidence of persistent,recurring summertime hypoxia in Green Bay,Lake Michigan

Six years (2009–2015) of temperature and dissolved oxygen profile data show hypoxic conditions are common in the bottom waters of southern Green Bay, Lake Michigan during the summer. Depleted oxygen concentrations (<5?mg?L^?1) affect nearly 70% of the 38 stations sampled representing an area of ~500–600?km². Stratification typically lasts 2+?months, from late June to early September, and some stations exhibit bottom water hypoxia (<2?mg?L^?1) at a frequency of nearly 25% when sampled during this period. A monitoring program initiated in 1986 by the Green Bay Metropolitan Sewerage District has provided a 23?year, recreational season record (May–September) of continuous (15?min interval) in situ bottom water oxygen and temperature measurements at the Entrance Light station of the Green Bay navigational channel. The duration of the hypoxic season ranges from 2?weeks to over 3?months at this shallow 7?m offshore site. This variability likely results from a combination of thermal stratification, oxygen consumption in deeper waters of the bay, and physical forcing mechanisms that drive cool, oxygen depleted, bottom waters on a southerly trajectory across this sensor. These data suggest the duration of hypoxic conditions may have increased during the stratified season in recent years. Hypoxia in the bay would also appear to be sensitive to relatively small changes in these forces, particularly changes in organic carbon loading and the duration of stratification.     

Understanding affective commitment in social virtual worlds: The role of cultural tightness

Understanding how cultural differences affect the formation of users’ affective commitment (ACO) to social virtual world (SVW) services is a critical but understudied issue. Using a novel, norm-based cultural approach, we introduce cultural tightness (CT) to capture cultural differences in users’ collective environment and postulate a model that investigates the contingent effects of CT on the relationships between ACO and its drivers. A two-level analysis of survey data gathered from 400 field users of an influential SVW service and secondary data of CT reveals that the effects of satisfaction, calculative commitment, and subjective norm on ACO are contingent on CT.     

Automated analysis of submicron particles by computer-controlled scanning electron microscopy

Automated analysis of submicron particles by computer-controlled scanning electron microscopy is generally possible. The minimum diameter of the detectable particles is dependent on the mean atomic number of the particles and the operating parameters of the scanning microscope. The main limitation with regard to particle size is set by the quality of the particle detection system, which generally is the backscatter electron detector. The accuracy of the results of the x-ray analyses is very often strongly affected by specimen damage, omnipresent especially for environmental particles even at low electron energies and probe currents. With the exception for light elements, the detection limit is approximately 1 wt%. Device-related limitations to automated analysis may be specimen drift and an unreliable autofocus function.     

Microbial community structure in activated sludge floc analysed by fluorescence in situ hybridization and its relation to floc stability

The efficiency of activated sludge treatment plants is dependent on the solid-liquid separation properties of the activated sludge. A critical parameter is the stability of the microbial flocs. Weak flocs deflocculate easily leaving increased concentrations of suspended solids in the effluent. The knowledge about how different bacteria are attached to the flocs and their influence on the bioflocculation is limited. In this study, the deflocculation of different phylogenetic groups of bacteria in activated sludge from a full scale plant was investigated. The experiments were carried out by using a shear method where the sludge flocs are deflocculated under controlled shear conditions. The degree of deflocculation was measured as increase in turbidity of the supernatant. Identification and quantification of the microbial community structure of both total activated sludge and deflocculated bacteria were conducted with group-specific gene probes for broad groups of bacteria (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria and Actinobacteria) and fluorescence in situ hybridization (FISH). The microbial community structure of the deflocculated bacteria was different compared to the total activated sludge with a higher abundance of Gammaproteobacteria in the supernatant indicating that different groups of bacteria are bound with different strength to the floc surface. The results show that the bacteria in the outer parts of the flocs are relatively loosely bound to the floc matrix and can be easily eroded from the surface when exposed to shear.     

Irradiation effects on AlGaN HFET devices and GaN layers

AlGaN/GaN heterostructure field effect transistors (HFETs) were irradiated with protons as well as carbon, oxygen, iron and krypton ions of high (68 and 120 MeV) and low (2 MeV) energy with fluences in the range from 1 × 10⁷ to 1 × 10¹³ cm^?2. High energy irradiation with protons, carbon and oxygen produced no degradation in devices while krypton irradiation at the fluence of 1 × 10¹⁰ cm^?2 resulted in a small reduction of 2% in the transconductance. Similarly, for GaN samples irradiated with protons, carbon and oxygen at high energy no changes were seen by XRD, PL and Hall effect, while changes in lattice constant and a reduction in PL intensity were observed after irradiation with high energy krypton. Low energy irradiation with carbon and oxygen at a fluence of 5 × 10¹⁰ cm^?2 results in small change in the device performance while remarkable changes in device characteristics are seen at a fluence of 1 × 10¹² cm^?2 for carbon, oxygen, iron and krypton irradiation. Similarly changes are also observed by XRD, PL and Hall effect for the thick GaN layer irradiated at the fluence of 1 × 10¹² cm^?2. The device results and GaN layer properties are strongly correlated.     

Monitoring diet effects via biofluids and their implications for metabolomics studies

The effect of diet on metabolites found in rat urine samples has been investigated using nuclear magnetic resonance (NMR) and a new ambient ionization mass spectrometry experiment, extractive electrospray ionization mass spectrometry (EESI-MS). Urine samples from rats with three different dietary regimens were readily distinguished using multivariate statistical analysis on metabolites detected by NMR and MS. To observe the effect of diet on metabolic pathways, metabolites related to specific pathways were also investigated using multivariate statistical analysis. Discrimination is increased by making observations on restricted compound sets. Changes in diet at 24-h intervals led to predictable changes in the spectral data. Principal component analysis was used to separate the rats into groups according to their different dietary regimens using the full NMR, EESI-MS data or restricted sets of peaks in the mass spectra corresponding only to metabolites found in the urea cycle and metabolism of amino groups pathway. By contrast, multivariate analysis of variance from the score plots showed that metabolites of purine metabolism obscure the classification relative to the full metabolite set. These results suggest that it may be possible to reduce the number of statistical variables used by monitoring the biochemical variability of particular pathways. It should also be possible by this procedure to reduce the effect of diet in the biofluid samples for such purposes as disease detection.     

Diode-pumped Nd:YAG laser with 38 W average power and user-selectable, flat-in-time subnanosecond pulses

A diode-pumped injection-seeded Nd:YAG laser system with an average output power of 38 W is described. The laser operates at 300 Hz with pulse energies up to 130 mJ. The temporal pulse shape is nominally flat in time and the pulse width is user selectable from 350 to 600 ps. In addition, the spatial profile of the beam is near top hat with contrast <10%.