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Leptin, insulin, and glucagon are involved in regulating glycaemia in vertebrates and play a role in the progression of obesity and type 2 diabetes. While mammals possess an ‘adipoinsular axis’ whereby insulin stimulates leptin release from adipocytes and leptin in turn feeds back on the pancreas to inhibit further insulin secretion, evidence of such an axis in non-mammalian vertebrates is unknown. We investigated the interactions between these glycaemic hormones and provide evidence for a leptin–insulin axis in a teleost fish, the tilapia. In the first study, we exposed hepatocytes to various concentrations of either insulin or glucagon to determine effects on leptin a (lepa) and then examined this in vivo with i.p. injections of both hormones. We also exposed isolated Brockmann bodies (pancreatic islets) to recombinant tilapia leptin A (rtLepA) and again followed this up with an i.p. injection to examine changes in insulin a and glucagon b. We found that glucagon increases lepa in vitroand in vivo, with the latter being 18-fold higher than saline-injected controls; however, the effects of rtLepA on glub were more variable. Insulin increased lepa by 2.5-fold in vitro and 70-fold in vivo, while rtLepA decreased insa at basal and increased it at high glucose concentrations. These data indicate that a leptin–insulin axis may be conserved among vertebrates and is thus essential for regulating nutrient balance but that the relationship is likely much more dynamic in teleosts as glycaemia is not as tightly regulated as it is in mammals.

Being a part of a social structure is key for survival and reproduction. Living with conspecifics boosts evolutionary fitness, by providing essential information about the environment. Nonetheless, studying neural mechanisms of social learning has not yet been established under laboratory conditions. To examine how socially passed information about the reward affects the behavior of individuals we used Eco-HAB, an automated system for tracing voluntary behavior of group-housed mice living under semi-naturalistic conditions. We show that a scent of a rewarded individual has profound effects on the conspecifics’ ability to find the reward in both familiar and novel environments. Importantly, the animals display clear and stable individual differences in social behavior. As a result, socially conveyed information has different effects on individual mice. Further, we show that disrupting neuronal plasticity in the prelimbic cortex with nanoparticles gradually releasing TIMP metallopeptidase inhibitor 1, disrupts animals’ social behavior and results in decreased ability to adapt to environmental changes. The experimental paradigm we developed can be further used to study neuronal mechanisms of social learning.

Environmental sex-determination (ESD) is the phenomenon by which environmental factors regulate sex-determination, typically occurring during a critical period of early development. Southern flounder (Paralichthys lethostigma) exhibit temperature-dependent sex-determination that appears to be restricted to the presumed XX female genotype with the extremes of temperature, both high and low, skewing sex ratios toward males. In order to evaluate other environmental factors that may influence sex-determination, we investigated the influence of background color and cortisol on sex-determination in southern flounder. Experiments involving three sets of tanks, each painted a different color, were conducted at different temperatures using southern flounder of mixed XX-XY genotype. The studies involved rearing juvenile southern flounder in either black, gray, or blue tanks and sex-determination was assessed by gonadal histology. In both studies, blue tanks showed significant male-biased sex ratios (95 and 75% male) compared with black and gray tanks. The stress corticosteroid cortisol may mediate sex-determining processes associated with environmental variables. Cortisol from the whole body was measured throughout the second experiment and fishes in blue tanks had higher levels of cortisol during the period of sex-determination. These data suggest that background color can be a cue for ESD, with blue acting as a stressor during the period of sex-determination, and ultimately producing male-skewed populations. In a separate study using XX populations of southern flounder, cortisol was applied at 0, 100, or 300 mg/kg of gelatin-coated feed. Fish were fed intermittently prior to, and just through, the period of sex-determination. Levels of gonadal P450 aromatase (cyp19a1) and forkhead transcription factor L2 (FoxL2) messenger RNA (mRNA) were measured by qRT-PCR as markers for differentiation into females. Müllerian-inhibiting substance mRNA was used as a marker of males' gonadal development. Control fish showed female-biased sex ratios approaching 100%, whereas treatment with 100 mg/kg cortisol produced 28.57% females and treatment with 300 mg/kg cortisol produced only 13.33% females. These results suggest that cortisol is a critical mediator of sex-determination in southern flounder by promoting masculinization. This linkage between the endocrine stress axis and conserved sex-determination pathways may provide a mechanism for adaptive modification of sex ratio in a spatially and temporally variable environment.

Leptin is an important cytokine for regulating energy homeostasis, however, relatively little is known about its function and control in teleost fishes or other ectotherms, particularly with regard to interactions with the growth hormone (GH)/insulin-like growth factors (IGFs) growth regulatory axis. Here we assessed the regulation of LepA, the dominant paralog in tilapia (Oreochromis mossambicus) and other teleosts under altered nutritional state, and evaluated how LepA might alter pituitary growth hormone (GH) and hepatic insulin-like growth factors (IGFs) that are known to be disparately regulated by metabolic state. Circulating LepA, and lepa and lepr gene expression increased after 3-weeks fasting and declined to control levels 10 days following refeeding. This pattern of leptin regulation by metabolic state is similar to that previously observed for pituitary GH and opposite that of hepatic GHR and/or IGF dynamics in tilapia and other fishes. We therefore evaluated if LepA might differentially regulate pituitary GH, and hepatic GH receptors (GHRs) and IGFs. Recombinant tilapia LepA (rtLepA) increased hepatic gene expression of igf-1, igf-2, ghr-1, and ghr-2 from isolated hepatocytes following 24 h incubation. Intraperitoneal rtLepA injection, on the other hand, stimulated hepatic igf-1, but had little effect on hepatic igf-2, ghr1, or ghr2 mRNA abundance. LepA suppressed GH accumulation and gh mRNA in pituitaries in vitro, but had no effect on GH release. We next sought to test if abolition of pituitary GH via hypophysectomy (Hx) affects the expression of hepatic lepa and lepr. Hypophysectomy significantly increases hepatic lepa mRNA abundance, while GH replacement in Hx fish restores lepa mRNA levels to that of sham controls. Leptin receptor (lepr) mRNA was unchanged by Hx. In in vitro hepatocyte incubations, GH inhibits lepa and lepr mRNA expression at low concentrations, while higher concentration stimulates lepa expression. Taken together, these findings indicate LepA gene expression and secretion increases with fasting, consistent with the hormones function in promoting energy expenditure during catabolic stress. It would also appear that LepA might play an important role in stimulating GHR and IGFs to potentially spare declines in these factors during catabolism. Evidence also suggests for the first time in teleosts that GH may exert important regulatory effects on hepatic LepA production, insofar as physiological levels(0.05–1 nM) suppresse lepa mRNA accumulation. Leptin A, may in turn exert negative feedback effects on basal GH mRNA abundance, but not secretion.

The intestinal epithelium forms a barrier protecting the organism from microbes and other proinflammatory stimuli. The integrity of this barrier and the proper response to infection requires precise regulation of powerful immune homing signals such as tumor necrosis factor (TNF). Dysregulation of TNF leads to inflammatory bowel diseases (IBD), but the mechanism controlling the expression of this potent cytokine and the events that trigger the onset of chronic inflammation are unknown. Here, we show that loss of function of the epigenetic regulator ubiquitin-like protein containing PHD and RING finger domains 1 (uhrf1) in zebrafish leads to a reduction in tnfa promoter methylation and the induction of tnfa expression in intestinal epithelial cells (IECs). The increase in IEC tnfa levels is microbe-dependent and results in IEC shedding and apoptosis, immune cell recruitment, and barrier dysfunction, consistent with chronic inflammation. Importantly, tnfa knockdown in uhrf1 mutants restores IEC morphology, reduces cell shedding, and improves barrier function. We propose that loss of epigenetic repression and TNF induction in the intestinal epithelium can lead to IBD onset.

Forward genetic approaches in zebrafish have provided invaluable information about developmental processes. However, the relative difficulty of mapping and isolating mutations has limited the number of new genetic screens. Recent improvements in the annotation of the zebrafish genome coupled to a reduction in sequencing costs prompted the development of whole genome and RNA sequencing approaches for gene discovery. Here we describe a whole exome sequencing (WES) approach that allows rapid and cost-effective identification of mutations. We used our WES methodology to isolate four mutations that cause kidney cysts; we identified novel alleles in two ciliary genes as well as two novel mutants. The WES approach described here does not require specialized infrastructure or training and is therefore widely accessible. This methodology should thus help facilitate genetic screens and expedite the identification of mutants that can inform basic biological processes and the causality of genetic disorders in humans.

The "Pi of the Sky" prototype apparatus observed prompt optical emission from extremely bright GRB080319B since the very beginning of the gamma emission. The burst occurred at redshift z=0.937 and set the record of optical luminosity reaching 5.3 mag. The position of the burst was observed before, during and after the explosion by several telescopes and unprecedented coverage of optical light curve has been achieved. The combination of these unique optical data with simultaneous gamma-ray observations provides a powerful diagnostic tool for the physics of the GRB explosion within seconds of its start. The "Pi of the Sky" prototype, working since 2004 in Las Campanas Observatory in Chile, consists of 2 cameras observing same 20x20 deg^2 fields in the sky with time resolution of 10 seconds. The prototype reacts to GCN alerts, but it has also its own algorithm for identification of optical flashes. The final system covering field of view of Swift or Fermi satellite will consist of 2 arrays of 16 cameras installed in a distance of about 100 km. The system is currently under construction. It will be a powerful tool for early optical observations of GRBs, allowing for optical observation of GRBs before, during and after the gamma emission. With the on-line data analysis in real time, it will identify short optical flashes autonomously and will be able to distribute this information among the community. In this contribution the general idea of the final version of the experiment and the most interesting results from the prototype are presented., 6 pages, 3 figures, to appear in Proc. of the 6th Huntsville Gamma-Ray Burst Symposium, eds. C. A. Meegan, N. Gehrels, and C. Kouveliotou (AIP Conf. Proc., New York)

The Pi of the Sky experiment searches for optical flashes of astronomical origin. Every night thousands of pictures of the sky (frames) are taken with CCD cameras. This article describes an algorithm designed for automatic estimation the quality of taken frames.© (2007) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

We have calculated the Wandzura-Wilczek contribution to the twist-3 part ofγ ∗ γ → 2π amplitude. It describes interaction of the longitudinally polarized virtualphoton with the real one, and it is suppressed by 1/Q, where Q 2 is the virtualityof the γ ∗ , as compared to the twist-2 contribution. We have found that, in theWandzura-Wilczek approximation, factorization applies to the twist-3 amplitude. 1 IntroductionHadron production in the reaction γ ∗ (q)γ(q ′ ) → hadron(s), q 2 = −Q 2 , has been a sub-ject of considerable interest in the QCD community from both experimental [1, 2] andtheoretical [3, 4, 5] points of view. Recently it has been proposed [6, 7] to investigate aprocess γ ∗ γ→ ππwhen the two pion state has a small invariant mass. Thanks to theQCD factorization theorem [8], in the leading twist approximation the amplitude can berepresented as a convolution of perturbatively calculable Wilson coefficients and new non-perturbative objects, the so-called two-pion distribution amplitudes (2πDA’s). They aregiven by matrix elements of twist-2 QCD string operators between vacuum and the two-pion state [7, 9]. Moreover, 2πDA’s can be related by the crossing symmetry to skewedparton distributions [10, 11] which recently have been subject of considerable interest.Later, NLO corrections to the leading twist amplitude have been calculated [12] and therecent thorough phenomenological analysis [13] have shown that experimental studies of2πproduction cross-section are possible with existing e

We have obtained a general solution of evolution equations for QCD twist-2 string operators in form of expansion over complete set of orthogonal eigenfunctions of evolution kernels in coordinate-space representation. In the leading logarithmic approximation the eigenfunctions can be determined using constraints imposed by conformal symmetry. Explicit formulae for the LO scale-dependence of quark and gluon twist-2 string operators are given.

We review the possibility to use the renormalons emerging in the perturbation series of the twist-2 part of the nonsinglet structure functions FL, F2, F3, and g1 to make approximate predictions for the magnitude of the appertaining twist-4 corrections., 10 pages, LaTeX, 4 figures, uses psfig and a41 (included) styles; Invited talk presented at the workshop 'Deep Inelastic Scattering off Polarized Targets: Theory Meets Experiment', Zeuthen, Germany, Sept. 1-5, 1997, two references added