Is there a downside to plant ecological services in the city? Influences of particulate matter on the two-spotted spider mite (Tetranychus urticae) foraging on the small-leaved lime in urban conditions.

The aim of this research was to examine how particulate matter (PM) pollution affects the life history of the two-spotted spider mite (TSSM), Tetranychus urticae (Trombidiformes: Tetranychidae), in modelled urban conditions. For this purpose, experimental populations of TSSM were cultured on the foliage of small-leaved lime (Tilia cordata) contaminated with PM at intensities corresponding to differing city zones such as a park, a busy road and an industrial area. The control samples in the study were washed, unpolluted leaves. The spider mite was selected as a model organism due to its cosmopolitan distribution, broad host spectrum, resistance to a variety of pesticides and food-intake mode involving cell-content sucking, while T. cordata is widely planted in cities and has demonstrated a considerable capability for PM capture. Data on the longevity and mortality of particular instars and on female fecundity at different pollution levels were collected and statistically evaluated. Concentrations of PM typical for roads and industrial city zones significantly reduced total female fecundity (avg. 53.9 and 55.9 eggs/female, respectively, vs 79.2 in control), which entailed a slower population increase, while the survival rate of particular developmental instars (P = 0.52) and fertility curves (P = 0.19) remained unchanged. The presence of PM caused physiological effects in the mites, despite the lack of direct consumption of the pollutant by adult and juvenile instars. Considering the incomparable resilience of TSSM to unfavourable environmental factors, it is predicted that the detrimental influence of PM on other representatives of urban arthropods may be even more severe. The results suggest that there is a need for further investigations into the ecological ramifications of air purification provided by urban green spaces.

Proglucagon signalling in the rat Dorsomedial Hypothalamus - Physiology and high-fat diet-mediated alterations.

A relatively new pharmacological target in obesity treatment has been the preproglucagon (PPG) signalling, predominantly with glucagon-like peptide (GLP) 1 receptor agonists. As far as the PPG role within the digestive system is well recognised, its actions in the brain remain understudied. Here, we investigated PPG signalling in the Dorsomedial Hypothalamus (DMH), a structure involved in feeding regulation and metabolism, using in situ hybridisation, electrophysiology, and immunohistochemistry. Our experiments were performed on animals fed both control, and high-fat diet (HFD), uncovering HFD-mediated alterations. First, sensitivity to exendin-4 (Exn4, a GLP1R agonist) was shown to increase under HFD, with a higher number of responsive neurons. The amplitude of the response to both Exn4 and oxyntomodulin (Oxm) was also altered, diminishing its relationship with the cells' spontaneous firing rate. Not only neuronal sensitivity, but also GLP1 presence, and therefore possibly release, was influenced by HFD. Immunofluorescent labelling of the GLP1 showed changes in its density depending on the metabolic state (fasted/fed), but this effect was eliminated by HFD feeding. Interestingly, these dietary differences were absent after a period of restricted feeding, allowing for an anticipation of the alternating metabolic states, which suggests possible prevention of such outcome.

Day/night Changes in the Dorsomedial Hypothalamus Firing Responses to Ghrelin are Modulated by High-fat Diet.

Dorsomedial hypothalamus (DMH) is a part of the feeding center involved in food intake and regulation of the metabolism. DMH neurons express many receptors for different metabolic cues which can modulate its network and influence animals' behaviour. One of the metabolic peptides deliveredto this structure is ghrelin, the only well-known hunger signal, produced mainly in the stomach. Diet-induced obesity is a physiological model of obesity widely used in research. Here we investigated how time-of-day and high-fat diet (HFD) affect neuronal networks and the sensitivity to the metabolic information received by the DMH. Our results indicate that even a short period of HFD (2-3 weeks) consumption can cause dysregulation of the DMH neuronal network, manifested as a disruption of the day/night pattern of basal activity and altered sensitivity to incoming information. We showed for the first time a day/night pattern of sensitivity to ghrelin in the DMH, with a higher level during the behaviorally active phase of animals. This day/night rhythm of sensitivity to ghrelin was reversed in HFD group, causing a stronger effect during the non-active phase. After prolongation of the HFD consumption to 7-8 weeks we observed an increase in the responsiveness to ghrelin, than during the short-term diet.

Enkephalin and neuropeptide-Y interaction in the intergeniculate leaflet network, a part of the mammalian biological clock.

The intergeniculate leaflet (IGL) is a flat thalamic nucleus implicated in the modulation of circadian rhythmicity. In rat, two main GABAergic subpopulations can be distinguished in the IGL: neurons synthesizing neuropeptide Y (NPY), which directly innervates the suprachiasmatic nuclei, and enkephalinergic cells, which connect contralaterally located leaflets. The aim of this study was to evaluate possible effects of inner IGL neurotransmitters on the spontaneous and synaptic activity of IGL neurons. The data presented in this article provide evidence that enkephalin, and not NPY, could act upon the majority of IGL neurons. Moreover, we investigated the type of opioid receptor activated by enkephalin and showed that the µ-receptor is functionally predominant in the IGL. The application of met-enkephalin not only robustly hyperpolarized IGL neurons (both putatively NPY-synthesizing and putatively enkephalinergic neurons), but it also was able to inhibit GABAergic and glutamatergic synaptic transmission. Based on this and previous studies, we hypothesize that IGL enkephalinergic neurons may act as powerful interneurons that inhibit themselves and NPY-synthesizing neurons, also in the contralaterally located IGL.

Orexins/hypocretins modulate the activity of NPY-positive and -negative neurons in the rat intergeniculate leaflet via OX1 and OX2 receptors.

Orexins/hypocretins (OXA and OXB) are two hypothalamic peptides involved in the regulation of many physiological processes including the sleep-wake cycle, food intake and arousal. The orexinergic system of the lateral hypothalamus is considered a non-specific peptidergic system, and its nerve fibers innervate numerous brain areas. Among many targets of orexinergic neurons is the intergeniculate leaflet (IGL) of the thalamus - a small but important structure of the mammalian biological clock. In rats, the IGL consists of GABAergic cells which also synthesize different neuropeptides. One group of neurons produces neuropeptide Y (NPY) and sends its axons to the master biological clock known as the suprachiasmatic nuclei. Another neuronal group produces enkephalin and is known to connect contralateral IGLs. This study evaluated the effects of orexins on identified IGL neurons revealing that 58% of the recorded neurons were sensitive to OXA (200nM) and OXB (200nM) administration. Both NPY-positive and -negative neurons were depolarized by these neuropeptides. Experiments using selective orexin receptor antagonists (SB-334867, 10µM and TCS-OX2-29, 10µM) suggested that both orexin receptors participate in the recorded OXA effects. In addition, IGL neurons were either directly depolarized by OXA or their activity was altered by changes in presynaptic inputs. We observed an increase of GABA release onto the investigated IGL neuron after OXA application, consistent with a presynaptic localization of the orexin receptors. An increase in miniature excitatory postsynaptic current frequency was not observed within the IGL. Our findings reinforce the connection between circadian clock physiology and the orexinergic system.

Bayesian QTL analyses using pedigreed families of an outcrossing species, with application to fruit firmness in apple.

KEY MESSAGE: Proof of concept of Bayesian integrated QTL analyses across pedigree-related families from breeding programs of an outbreeding species. Results include QTL confidence intervals, individuals' genotype probabilities and genomic breeding values. Bayesian QTL linkage mapping approaches offer the flexibility to study multiple full sib families with known pedigrees simultaneously. Such a joint analysis increases the probability of detecting these quantitative trait loci (QTL) and provide insight of the magnitude of QTL across different genetic backgrounds. Here, we present an improved Bayesian multi-QTL pedigree-based approach on an outcrossing species using progenies with different (complex) genetic relationships. Different modeling assumptions were studied in the QTL analyses, i.e., the a priori expected number of QTL varied and polygenic effects were considered. The inferences include number of QTL, additive QTL effect sizes and supporting credible intervals, posterior probabilities of QTL genotypes for all individuals in the dataset, and QTL-based as well as genome-wide breeding values. All these features have been implemented in the FlexQTL(™) software. We analyzed fruit firmness in a large apple dataset that comprised 1,347 individuals forming 27 full sib families and their known ancestral pedigrees, with genotypes for 87 SSR markers on 17 chromosomes. We report strong or positive evidence for 14 QTL for fruit firmness on eight chromosomes, validating our approach as several of these QTL were reported previously, though dispersed over a series of studies based on single mapping populations. Interpretation of linked QTL was possible via individuals' QTL genotypes. The correlation between the genomic breeding values and phenotypes was on average 90 %, but varied with the number of detected QTL in a family. The detailed posterior knowledge on QTL of potential parents is critical for the efficiency of marker-assisted breeding.

Modified multi-element synthetic transmit aperture method for ultrasound imaging: A tissue phantom study.

The paper presents the modified multi-element synthetic transmit aperture (MSTA) method for ultrasound imaging. It is based on coherent summation of RF echo signals with apodization weights taking into account the finite size of the transmit subaperture and of the receive element. The work presents extension of the previous study where the modified synthetic transmit aperture (STA) method was considered and verified [1]. In the case of MSTA algorithm the apodization weights were calculated for each imaging point and all combinations of the transmit subaperture and receive element using their angular directivity functions (ADFs). The ADFs were obtained from the exact solution of the corresponding mixed boundary-value problem for periodic baffle system modeling the transducer array. Performance of the developed method was tested using Field II simulated synthetic aperture data of point reflectors for 4MHz 128-element transducer array with 0.3mm pitch and 0.02mm kerf to estimate the visualization depth and lateral resolution. Also experimentally determined data of the tissue-mimicking phantom (Dansk Fantom Service, model 571) obtained using 128 elements, 4MHz, linear transducer array (model L14-5/38) and Ultrasonix SonixTOUCH Research platform were used for qualitative assessment of imaging contrast improvement. Comparison of the results obtained by the modified and conventional MSTA algorithms indicated 15dB improvement of the noise reduction in the vicinity of transducer's surface (1mm depth), and concurrent increase in the visualization depth (86% augment of the scattered amplitude at the depth of 90mm). However, this increase was achieved at the expense of minor degradation of the lateral resolution of approximately 8% at the depth of 50mm and 5% at the depth of 90mm.

Differential firing pattern and response to lighting conditions of rat intergeniculate leaflet neurons projecting to suprachiasmatic nucleus or contralateral intergeniculate leaflet.

The intergeniculate leaflet (IGL) of the lateral geniculate body in the rat is a population of GABAergic neurons that can be divided into two, anatomically and neurochemically distinct populations. One population comprises neuropeptide-Y (NPY)-positive neurons that form the geniculohypothalamic tract innervating the suprachiasmatic nuclei (SCN) and the other population comprises enkephalin-positive (ENK) neurons giving rise to the geniculo-geniculate tract innervating the contralateral IGL (cIGL). Previous electrophysiological studies have observed various patterns of firing and different responses to changes in lighting conditions of IGL neurons in vitro and in vivo. The aim of the present study was to determine if these distinct properties could be ascribed to differentially projecting IGL neurons. Neuron activity was recorded extracellularly in the IGL of anaesthetised rats under different lighting conditions (i.e. light/dark). Antidromic activation was used to identify recorded cells as projecting to the SCN or the contralateral IGL. All IGL neurons identified as projecting to the contralateral IGL displayed infra-slow oscillatory activity (ISO; i.e. slow rhythmic bursts of action potentials). ISO of these neurons was sustained in the light and was diminished in the darkness. In contrast, all IGL neurons identified as projecting to the SCN displayed a low level of firing in the light and a majority of these cells increased firing in the darkness. All IGL neurons projecting to the SCN were characterised by an irregular pattern of firing in the light and dark. These data are the first to demonstrate that differentially projecting rat intergeniculate leaflet neurons are characterised by distinct firing patterns and opposite responses to light and dark conditions.

Depolarization of the intergeniculate leaflet neurons by serotonin - in vitro study.

The intergeniculate leaflet of the thalamus (IGL) is a part of the mammalian biological clock which integrates photic and non-photic information and conveys it to the master biological clock - suprachiasmatic nuclei (SCN). One of the non-photic cues is delivered by the serotoninergic projection from dorsal raphe nucleus. In vitro electrophysiological recordings were performed from single IGL neurons using whole-cell patch clamp technique. We investigated the influence of serotonin (serotonin creatinine sulfate complex, 5HT) on 'spontaneous' neuronal activity in this structure. In most of recorded cells, 5-HT caused significant increases in firing rate. In majority of cases the effect was presynaptic. However, in some cases we observed postsynaptic depolarization. To our knowledge, depolarizing influence of 5HT on the single neurons in the IGL has been shown here for the first time.

The effectiveness of anticellulite treatment using tripolar radiofrequency monitored by classic and high-frequency ultrasound.

BACKGROUND: Cellulite affects nearly 85% of the female population. Given the size of the phenomenon, we are continuously looking for effective ways to reduce cellulite. Reliable monitoring of anticellulite treatment remains a problem. OBJECTIVE: The main aim of the study was to evaluate the effectiveness of anticellulite treatment carried out using radiofrequency (RF), which was monitored by classical and high-frequency ultrasound. METHODS: Twenty-eight women underwent anticellulite treatment using RF, 17 women were in the placebo group. The therapy was monitored by classical and high-frequency ultrasound. The examinations evaluated the thickness of the epidermal echo, dermis thickness, dermis echogenicity, the length of the subcutaneous tissue bands growing into the dermis, the presence or absence of oedema, the thickness of subcutaneous tissue as well as thigh circumference and the stage of cellulite (according to the Nürnberger-Müller scale). RESULTS: Cellulite was reduced in 89.286% of the women who underwent RF treatment. After the therapy, the following observations were made: a decrease in the thickness of the dermis and subcutaneous tissue, an increase in echogenicity reflecting on the increase in the number of collagen fibres, decreased subcutaneous tissue growing into bands in the dermis, and the reduction of oedema. In the placebo group, no statistically significant changes of the above parameters were observed. CONCLUSION: Radiofrequency enables cellulite reduction. A crucial aspect is proper monitoring of the progress of such therapy, which ultrasound allows.

Effect of single bout of maximal exercise on plasma antioxidant status and paraoxonase activity in young sportsmen.

The purpose of this study was to elucidate the participation of plasma PON1 (paraoxonase activity [PON] and arylesterase activity [ARE]) in antioxidant defense in response to a single bout of maximal exercise. PON, ARE, lipid profile, lipid peroxidation (thiobarbituric acid reactive substances [TBARS]), total antioxidant status (ferric reducing ability of plasma [FRAP]), concentration of uric acid [UA], and total bilirubin (TBil) were determined in the plasma before, at the bout and 2 h after maximal exercise on a treadmill in young sportsmen. Chosen physiological parameters also were controlled during maximal exercise. Following maximal exercise, the unaltered level of TBARS and increased FRAP were registered. ARE increment was the highest (37.6%) of all measured variables but lasted for a short time. UA increment was lower than ARE but long-lasting and correlated with FRAP. PON activity increment was associated with the combined effect of body weight, lean, body mass index (BMI) and basal metabolic rate (BMR). We conclude that PON1 is a co-factor of the first line of antioxidant defense during maximal exercise. Its activity is associated with body composition and not the physical fitness of the subjects.

Electrophysiology and pharmacology of the optic input to the rat intergeniculate leaflet in vitro.

The mammalian intergeniculate leaflet (IGL) of the thalamus is a neuronal element of the circadian timing system, which receives direct photic input from the retina. The purpose of this study was to analyze responses of rat IGL neurons in vitro to optic tract stimulation and to identify neurotransmitters released from the terminals of retinal ganglion cells in this structure. Following optic tract stimulation, most of the responding IGL cells were excited and only a minority of them were inhibited. Neurons showing the excitatory response were tested in the presence of AP-5, a selective antagonist of NMDA receptors. In most cases the responses were only partially inhibited by the presence of AP-5. Complete disappearance of excitatory responses was achieved by adding CNQX, an AMPA/kainate receptor-selective antagonist, to the standard incubation fluid. Inhibitory responses were blocked or considerably attenuated in the presence of bicuculline, a GABA(A) receptor antagonist, in the ACSF. This study demonstrated that glutamate is the main neurotransmitter mediating optic tract input to the IGL, acting mainly via non-NMDA ionotropic receptors. It was also shown that NMDA and GABA(A) receptors are involved in passing photic input to the IGL, albeit to a much lesser extent.

Morphology and CO adsorption on platinum supported on thin Fe(3)O(4)(111) films.

Nucleation, growth and thermal stability of Pt particles supported on well ordered Fe(3)O(4)(111) thin films grown on Pt(111) were studied by scanning tunnelling microscopy (STM) and temperature programmed desorption (TPD) of CO. STM studies showed that Pt grows through the formation of single-layer islands that coalesce at high coverage. Vacuum annealing at 600 K caused Pt sintering and the formation of extended two-dimensional (2D) islands one and two layers in thickness at sub-monolayer coverage. Well faceted, three-dimensional (3D) Pt nanoparticles formed by annealing to temperatures above 800 K were encapsulated by an FeO(111) monolayer. These results were rationalized in terms of the high adhesion energy for Pt on iron oxide surfaces. CO TPD studies showed that 2D structures, formed at 600 K, exhibit much lower CO adsorption capacity as compared to the Pt(111) single crystal surface. This effect has been tentatively assigned to lattice expansion in the Pt 2D islands leading to weakening of the Pt-CO bond due to reduction of the [Formula: see text] back-donation.

The responsiveness of the rat intergeniculate leaflet neurons to glutamatergic agonists.

The intergeniculate leaflet (IGL) has been shown to be a functional constituent of the circadian timing system. The IGL receives a monosynaptic input from the retina and is known to mediate some of the effects of light on the circadian clock. In the majority of retinal ganglion cells, glutamate functions as an excitatory neurotransmitter. The effect of monosodium glutamate and N-methyl-D-aspartate (NMDA), on the extracellularly recorded discharge activity of IGL neurons was studied in vitro. The application of monosodium glutamate induced either an excitatory, a biphasic or an inhibitory response. Application of NMDA induced an excitatory response in the majority of tested neurons. To determine the role of NMDA receptors in the response to glutamate application, the selective antagonist of NMDA receptors- AP-5, was applied to the incubation medium. The presence of AP-5 reduced the response of the IGL cells to focal application of glutamate and completely blocked their responsiveness to NMDA. To clarify whether GABAergic interneurons are involved in mediation of the inhibitory effects of glutamate, we repeated our experiments in the presence of bicuculline in the incubation medium. Since bicuculline did not influence the observed inhibitory effects, the involvement of GABAA receptors was excluded. The present study provides the first electrophysiological evidence that neurons in the rat IGL, respond to glutamate probably through NMDA receptors. However, our results also suggest that other types of glutamate receptors may play an additional role in mediating the action of this excitatory amino acid on the IGL neurons.

The serotonergic inhibition of slowly bursting cells in the intergeniculate leaflet of the rat.

Electrophysiological studies combined with local neurotoxic lesions were conducted on anaesthetized rats in order to determine whether the dorsal raphe nucleus (DRN) inhibits the intergeniculate leaflet (IGL) of the lateral geniculate nucleus by means of innervation by serotonin-containing fibres. In the control animals, electrical stimulation of the DRN induced the long-latency and long-lasting inhibition of the neuronal firing of the IGL cells that are characterized by rhythmic, slow-bursting activity in light conditions. The electrical destruction of the DRN resulted in an increase in the firing rate of the recorded IGL cells, whilst at the same time not affecting the rhythmic, bursting pattern of the activity. In the second group of animals, local neurotoxic lesion of serotonergic fibres was performed by injection of the toxin 5,7-dihydroxytryptamine into the IGL. After 10 days of postoperative recovery, electrophysiological experiments were performed on the toxin-treated rats. In these animals, electrical stimulation as well as electrical lesion of the DRN did not induce any change in the firing of the slowly bursting cells in the 5,7-dihydroxytryptamine-injected IGL. The results obtained provide evidence that inhibition of the IGL slowly bursting cells, by innervation from the dorsal raphe, is mediated by the release of serotonin. Furthermore, the observed serotonergic inhibition of the light-dependent activity of slowly bursting cells can contribute to the neuronal mechanism gating the information that flows through this nucleus to the vestibular, visuomotor, circadian and sleep/arousal systems, with which the IGL is strongly interconnected.

Epidemiology of ARDS.

For decades the incidence of the acute respiratory distress syndrome (ARDS) has been unclear. A first effort to tackle the problem was undertaken by the National Heart and Lung Task Force who, in 1972, provided the scientific community with an arguable estimate of 75 cases per 100,000 inhabitants/year for the USA. Nearly 20 years later, the first population based studies yielded figures of 1.5-4.5 cases per 100 000 inhabitants/year in Europe. Epidemiologic research became much more focused when in 1992 the new ARDS and acute lung injury (ALI) definitions of the American-European Consensus Conference became available. In subsequent studies in which these refined definitions were used, incidence figures ranging from 13-23 cases per 100,000 inhabitants/year for ARDS and 18 cases per 100,000 inhabitants/year for ALI were reported. Latest results from a high-class epidemiological study conducted in Seattle, suggested ARDS/ALI incidence figures of 59 and 79 cases per 100,000 inhabitants/year, respectively. These new figures, similar to the one proposed in 1972, led to a paradigm shift: ARDS and ALI are no longer viewed as rare syndromes but as widespread diseases with a massive socio-economic impact that is comparable with the burden from breast cancer, AIDS, asthma or myocardial infarction.

Comparison of sound fields generated by different coded excitations--experimental results.

This work reports the results of measurements of spatial distributions of ultrasound fields obtained from five energizing schemes. Three different codes, namely, chirp signal and two sinusoidal sequences were investigated. The sequences were phase modulated with 13 bits Barker code and 16 bits Golay complementary codes. Moreover, two reference signals generated as two and sixteen cycle sine tone bursts were examined. Planar, 50% (fractional) bandwidth, 15 mm diameter source transducer operating at 2 MHz center frequency was used in all measurements. The experimental data were collected using computerized scanning system and recorded using wideband, PVDF membrane hydrophone (Sonora 804). The measured echoes were compressed, so the complete pressure field in the investigated location before and after compression could be compared. In addition to a priori anticipated increase in the signal to noise ratio (SNR) for the decoded pressure fields, the results indicated differences in the pressure amplitude levels, directivity patterns, and the axial distance at which the maximum pressure amplitude was recorded. It was found that the directivity patterns of non-compressed fields exhibited shapes similar to the patterns characteristic for sinusoidal excitation having relatively long time duration. In contrast, the patterns corresponding to compressed fields resembled those produced by brief, wideband pulses. This was particularly visible in the case of binary sequences. The location of the maximum pressure amplitude measured in the 2 MHz field shifted towards the source by 15 mm and 25 mm for Barker code and Golay code, respectively. The results of this work may be applicable in the development of new coded excitation schemes. They could also be helpful in optimizing the design of imaging transducers employed in ultrasound systems designed for coded excitation. Finally, they could shed additional light on the relationship between the spatial field distribution and achievable image quality and in this way facilitate optimization of the images obtained using coded systems.

Identification and quantification of aerosol polar oxygenated compounds bearing carboxylic or hydroxyl groups. 2. Organic tracer compounds from monoterpenes.

In this study, a comparison is made of polar organic compounds found in the field with those produced in secondary organic aerosol from laboratory irradiations of natural hydrocarbons and oxides of nitrogen (NOx). The field samples comprised atmospheric particulate matter (PM2.5) collected at Research Triangle Park (RTP), NC, during the summer of 2003, and the laboratory samples originated from the photooxidation of the following monoterpenes: alpha-pinene, beta-pinene, and d-limonene. To determine the structural characteristics of the polar compounds, the filter samples were solvent extracted and derivatized using a technique based on single and multistep derivatizations. The resulting compound derivatives were analyzed by GC-MS in the methane-Cl and El modes. In addition to previously reported biogenic oxidation products (pinic acid, pinonic acid, norpinic acid, nopinone, and pinonaldehyde), seven multifunctional organic compounds were found in both field and laboratory samples. These compounds, which are proposed as possible atmospheric tracers for secondary organic aerosol from monoterpenes, were consistent with the following identifications: 3-isopropyl pentanedioic acid; 3-acetyl pentanedioic acid; 3-carboxy heptanedioic acid; 3-acetyl hexanedioic acid; 2-isopropyl-1,2-dihydroxybutanol; 4-isopropyl-2,4-dihydroxyhexanol; and 3-(2-hydroxy-ethyl)-2,2-dimethyl-cyclobutane carboxylic acid. Initial attempts have been made to quantify the concentrations of these tracer compounds on the basis of surrogate compound calibrations. The occurrence of these compounds in both laboratory and field measurements suggests that secondary organic aerosol originating from biogenic hydrocarbons are contributing to the regional aerosol burden in the southeastern United States. Several of these compounds also appear to contribute to the global aerosol burden in that they have also been identified in Europe and Brazil.

Identification and quantification of aerosol polar oxygenated compounds bearing carboxylic or hydroxyl groups. 1. Method development.

In this study, a new analytical technique was developed for the identification and quantification of multifunctional compounds containing simultaneously at least one hydroxyl or one carboxylic group, or both. This technique is based on derivatizing first the carboxylic group(s) of the multifunctional compound using an alcohol (e.g., methanol, 1-butanol) in the presence of a relatively strong Lewis acid (BF3) as a catalyst. This esterification reaction quickly and quantitatively converts carboxylic acids to their ester forms. The second step is based on silylation of the ester compounds using bis(trimethylsilyl) trifluoroacetamide (BSTFA) as the derivatizing agent. For compounds bearing ketone groups in addition to carboxylic and hydroxyl groups, a third step was used based on PFBHA derivatization of the carbonyls. Different parameters including temperature, reaction time, and effect due to artifacts were optimized. A GC/MS in EI and in methane-CI mode was used for the analysis of these compounds. The new approach was tested on a number of multifunctional compounds. The interpretation of their EI (70 eV) and CI mass spectra shows that critical information is gained leading to unambiguous identification of unknown compounds. For example, when derivatized only with BF(3)-methanol, their mass spectra comprise primary ions at m/z M.+ + 1, M.+ + 29, and M.+ - 31 for compounds bearing only carboxylic groups and M.+ + 1, M.+ + 29, M.+ - 31, and M+. - 17 for those bearing hydroxyl and carboxylic groups. However, when a second derivatization (BSTFA) was used, compounds bearing hydroxyl and carboxylic groups simultaneously show, in addition to the ions observed before, ions at m/z M.+ + 73, M.+ - 15, M.+ - 59, M.+ - 75, M.+ - 89, and 73. To the best of our knowledge, this technique describes systematically for the first time a method for identifying multifunctional oxygenated compounds containing simultaneously one or more hydroxyl and carboxylic acid groups.

Blockade of GABAA receptors disrupts isoperiodic neuronal oscillations in the intergeniculate leaflet of the rat.

The intergeniculate leaflet of the thalamus is, besides the suprachiasmatic nucleus of the hypothalamus, the other important neuronal element of the mammalian biological clock. The extracellularly recorded activity of neurons constituting the intergeniculate leaflet, recorded in vivo, is characterized by distinct, very regular ultradian oscillations. The majority of neurons in the circadian timing system are GABAergic. Many, if not all, neurons of the suprachiasmatic nucleus and intergeniculate leaflet contain GABA. In the present study we examined the effects of the GABA(A) receptor antagonist bicuculline and the chloride channel blocker picrotoxin on isoperiodic neuronal oscillations in the intergeniculate leaflet of rats. We recorded extracellular multiple-unit neuronal activity from the intergeniculate leaflet of anesthetized rats. During the recording of isoperiodic oscillations, bicuculline or picrotoxin were stereotaxically injected at different concentrations into the lateral ventricle of rat brain. In all the experiments, injection of GABA(A) receptor antagonists transiently disrupted the isoperiodic phasic discharge recorded from the intergeniculate leaflet. These data suggest that GABA(A) receptors are involved in the generation of ultradian rhythmical neuronal oscillations in rat intergeniculate leaflet.