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.

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.

Particulate matter hinders the development and reproduction of predatory mites of Euseius finlandicus (Acariformes: Phytoseiidae).

The foliage of the small-leaved lime (Tilia cordata) is characterised by the nerve axils being grown by non-glandular trichomes, which trait contributes to the enhanced retention of the particulate matter (PM). This fact may disturb the ecological service of T. cordata related to the structure of its leaves, which is to provide acarodomatia (micro-shelters) for the predatory mites of the Phytoseiidae family. Phytoseiids are natural enemies of a variety of plant pests, widely applied in integrated pest management (IPM). Their occurrence is largely related to acarodomatia in which these mites hide, feed, reproduce, and develop. For the first time, the influence of PM deposition within spaces typically occupied by phytoseiids is investigated. Experimental populations of Euseius finlandicus were reared on T. cordata leaves in the progressive PM-pollution. The results showed that the values of life table parameters of the predator depended significantly on the level of PM deposition on leaves. Contrary to clean leaves from the control, the medium and high contamination intensities significantly reduced the daily (by 47% and 70%, respectively) and the total fecundity (by 62% and 77%, respectively) of females which, in turn, resulted in a decreased net reproductive rate (by 67% and 81%, respectively), intrinsic rate of increase (by 40% and 55%, respectively) and finite rate of increase (by 8% and 10%, respectively) of E. finlandicus. The pre-ovipositional period was prolonged, while the oviposition duration was shortened and the mites matured longer. In high pollution level the mortality of phytoseiids was boosted by 19% and some females were observed with pollutant lumps adhered to the idiosoma. Also, significant shares of juvenile forms (13%) and adult females (25%) made attempts to escape from highly contaminated experimental arenas. The implications of PM retention on the shelter vegetation are discussed in the context of IPM and ecological services.

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.

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.

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.

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.

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.

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.

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.

Magnetic resonance assessment of cerebral perfusion in depressed cardiac patients: preliminary findings.

OBJECTIVE: The authors investigated the links between depression, cardiac disease, and microcirculatory cerebral blood flow (CBF). METHOD: A magnetic resonance imaging technique based on arterial spin tagging was used to estimate microcirculatory CBF in depressed (N = 5) and comparison (N = 14) elderly subjects with coronary artery disease. Signal intensity ratios corresponding to relative microcirculatory CBF were calculated for four regions on two axial images through the upper and lower halves of the lateral ventricles. RESULTS: On the superior image, estimates of microcirculatory CBF were statistically significantly lower on the left side in the depressed subjects than in the nondepressed group. When the ratios in the superior and inferior images were averaged, the depressed subjects had lower values for both left periventricular regions of interest and the parietal region. CONCLUSIONS: Despite the small study group and indirect estimates of blood flow, these preliminary findings suggest that a relative cerebral hypoperfusion may underlie depression in elderly cardiac patients.

Echographic monitoring of response of extraocular muscles to irradiation in Graves' ophthalmopathy.

PURPOSE: Confirmation of the efficacy of orbital irradiation in Graves' ophthalmopathy is needed due to the unpredictable natural history of the disease, the variation in individual clinical presentations, the contribution of other simultaneous treatments, and the lack of controlled studies using objective criteria to classify and assess response over time. Orbital echography before and at select intervals following orbital irradiation is proposed as an objective parameter of tissue response to orbital irradiation over time. METHODS AND MATERIALS: From January, 1983 to September, 1993, 55 patients with progressive Graves' ophthalmopathy underwent 20 Gy retrobulbar irradiation. On retrospective review, standardized orbital echography was performed randomly prior to irradiation in 37 of the 55 patients to assess the acoustic characteristics of the extraocular muscles and to quantitate their individual and summed diameters. Twenty-one patients had at least one follow-up echographic evaluation at random intervals of 0 to 27.5 months following completion of irradiation. Twelve patients received steroids before or during irradiation, which were tapered in proximity to completion of radiation. Follow-up ranged from 2 to 65 months with the majority followed at least 6 months (18 patients). RESULTS: Of the 21 patients with serial studies, 18 showed an interval decrease in individual and summed muscle size over time and return of symmetry. Interval improvement was documented as early as the 1 month follow-up study, with continued improvement seen during the 3-9-month studies, with stability typically achieved within 12 months. One patient had further changes between the 21 and 27.5 month follow-up studies. Exacerbation of disease was, however, echographically demonstrated in three patients at 6.5, 8.5, and 13 months. Follow-up studies in two of these patients again revealed improvement, one following tapered steroids. The third patient required orbital decompression. CONCLUSION: Objective parameters of response are needed to document both the immediate and long-term outcome of orbital irradiation on the course of Graves' ophthalmopathy and confirm its efficacy. Serial echography is proposed as a new technique for providing parameters to judge response.

Electro-oculography in autosomal dominant vitreoretinochoroidopathy.

Thirteen members of a family presumed to be harboring the gene for autosomal dominant vitreoretinochoroidopathy were examined. In four affected members, electro-oculography demonstrated marked reduction of the Arden ratio (range, 1.1 to 1.5; normal, > or = 1.8), despite electroretinographic evidence of mildly affected rod function and normal cone function. These findings suggested that a diffuse disturbance of the photoreceptor-pigment epithelium complex may have been present prior to wide-spread loss of photoreceptor function in the affected members of this family. As in previously described families, the pattern of inheritance appeared consistent with autosomal dominance.

Echographic diagnosis of anterior hyaloidal fibrovascular proliferation.

High-resolution contact B-scan echographic imaging of the ciliary body and peripheral retina was performed on five eyes with anterior hyaloidal fibrovascular proliferation and media opacity by means of a wide (58 degrees) scanning arc. This technique determined the circumferential and anteroposterior extent of peripheral traction retinal detachment associated with anterior hyaloidal fibrovascular proliferation, which correlated highly with findings at subsequent vitrectomy. Final visual acuity of 20/400 or better was achieved in the two eyes (50%) with more limited peripheral traction detachment. In the presence of media opacity, anterior echographic imaging may allow early detection of traction retinal detachment associated with anterior hyaloidal fibrovascular proliferation and may be useful in characterizing the severity of this condition.

Echographic verification of radioactive plaque position in the treatment of melanomas.

Visualization of the position of an episcleral plaque in relation to the underlying choroidal melanoma during the early radiation treatment period reconfirms accurate plaque location and may help assure optimal treatment. We used high-resolution B-scan echography to confirm the position of the episcleral plaque in 16 consecutive cases of choroidal melanoma. The technique is most useful for well-defined tumors as small as 3 mm in height whose anterior border does not extend beyond the ora serrata.

Titanium retinal tacks with pneumatic insertion. Histologic evaluation in rabbits.

Titanium retinal tacks were implanted through the formed vitreous into the posterior retinas of rabbits and evaluated by electroretinographic testing and by light microscopy at intervals after implantation. Tritiated thymidine autoradiography was used to evaluate the proliferation of ocular tissues in response to tack insertion. Within one month after implantation, the retinal tacks were surrounded by scar tissue produced by the proliferation of retinal glial cells and choroidal and scleral connective tissue cells. The tissue response was focal and there was no excessive fibrous tissue or vascular proliferation or any indication of iris neovascularization for six months. The observations of firm retinal adherence and low tissue reactivity suggest that titanium tacks can be used to secure the retina during retinal attachment surgery and can remain permanently in the eye.

Methanol poisoning. A rodent model with structural and functional evidence for retinal involvement.

Methanol ingestion can lead to visual impairment, central nervous system dysfunction, or death. The extent of ocular involvement has been difficult to determine because the toxicity is restricted to humans and nonhuman primates due to species differences in methanol metabolism. A rodent model of methanol toxicity recently developed by us was used to evaluate retinal dysfunction in methanol poisoning. Formic acidemia and visual toxic reactions developed in methanol-intoxicated rats. Electroretinographic analysis indicated a significant early deficit in b-wave amplitude followed by a temporally delayed, lesser reduction in a-wave amplitude. Histologic evaluation of the eyes 60 hours after methanol administration revealed generalized retinal edema and vacuolation in the photoreceptors and retinal pigment epithelium. Ultrastructural examination showed swelling and disruption of the mitochondria in photoreceptor inner segments, optic nerve, and the retinal pigment epithelium. These studies document direct retinal involvement in this nonprimate model of methanol toxicity.

Ultradian rhythmic neuronal oscillation in the intergeniculate leaflet.

Our paper is the first to describe ultradian rhythmic neuronal oscillation in the intergeniculate leaflet (IGL) of the rat. We recorded a multiple-unit neuronal activity (MUA) from dorsal to ventral parts of the lateral geniculate nucleus (LGN) in anaesthetized rats. In all the subdivisions of the lateral geniculate complex we observed spontaneous irregular firing rates of cells. However only at the anatomical localisation of the IGL, after the light was on, those responses exhibited burst firing with a constant interburst interval, which lasted several hours until the light was off. The duration of that rhythmic oscillation obtained by means of Fourier's analysis was approximately 124 s. To date we have not had sufficient data to discuss possible mechanisms of this neuronal rhythmicity. We can only conclude that light is the most important stimulus not only for suprachiasmatic nuclei (SCN), but also for the IGL. On the other hand, we can neither exclude nor confirm that in order to evoke ultradian rhythmical oscillation in the IGL, in addition to light also non-photic information is necessary.

The effect of muscarinic and nicotinic ACh antagonist on the facilitation of rat visual cortical responses.

Combined blockade of both muscarinic and nicotinic receptors fully eliminated the effect of reticular facilitation in rat visual cortex. However, this effect lasted only 2 h, what can suggest that there may be other excitatory input to the cholinergic neurones in rat visual cortex which is activated after blockade of cholinergic transmission.

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.