Functional Inactivation of Drosophila GCK Orthologs Causes Genomic Instability and Oxidative Stress in a Fly Model of MODY-2

Maturity-onset diabetes of the young (MODY) type 2 is caused by heterozygous inactivating mutations in the gene encoding glucokinase (GCK), a pivotal enzyme for glucose homeostasis. In the pancreas GCK regulates insulin secretion, while in the liver it promotes glucose utilization and storage. We showed that silencing the Drosophila GCK orthologs Hex-A and Hex-C results in a MODY-2-like hyperglycemia. Targeted knock-down revealed that Hex-A is expressed in insulin producing cells (IPCs) whereas Hex-C is specifically expressed in the fat body. We showed that Hex-A is essential for insulin secretion and it is required for Hex-C expression. Reduced levels of either Hex-A or Hex-C resulted in chromosome aberrations (CABs), together with an increased production of advanced glycation end-products (AGEs) and reactive oxygen species (ROS). This result suggests that CABs, in GCK depleted cells, are likely due to hyperglycemia, which produces oxidative stress through AGE metabolism. In agreement with this hypothesis, treating GCK-depleted larvae with the antioxidant vitamin B6 rescued CABs, whereas the treatment with a B6 inhibitor enhanced genomic instability. Although MODY-2 rarely produces complications, our data revealed the possibility that MODY-2 impacts genome integrity.     

In Vivo Silencing of Genes Coding for dTip60 Chromatin Remodeling Complex Subunits Affects Polytene Chromosome Organization and Proper Development in Drosophila melanogaster

Chromatin organization is developmentally regulated by epigenetic changes mediated by histone-modifying enzymes and chromatin remodeling complexes. In Drosophila melanogaster, the Tip60 chromatin remodeling complex (dTip60) play roles in chromatin regulation, which are shared by evolutionarily-related complexes identified in animal and plants. Recently, it was found that most subunits previously assigned to the dTip60 complex are shared by two related complexes, DOM-A.C and DOM-B.C, defined by DOM-A and DOM-B isoforms, respectively. In this work, we combined classical genetics, cell biology, and reverse genetics approaches to further investigate the biological roles played during Drosophila melanogaster development by a number of subunits originally assigned to the dTip60 complex.     

Searching for motifs in the behaviour of larval Drosophila melanogaster and Caenorhabditis elegans reveals continuity between behavioural states

We present a novel method for the unsupervised discovery of behavioural motifs in larval Drosophila melanogaster and Caenorhabditis elegans. A motif is defined as a particular sequence of postures that recurs frequently. The animal''s changing posture is represented by an eigenshape time series, and we look for motifs in this time series. To find motifs, the eigenshape time series is segmented, and the segments clustered using spline regression. Unlike previous approaches, our method can classify sequences of unequal duration as the same motif. The behavioural motifs are used as the basis of a probabilistic behavioural annotator, the eigenshape annotator (ESA). Probabilistic annotation avoids rigid threshold values and allows classification uncertainty to be quantified. We apply eigenshape annotation to both larval Drosophila and C. elegans and produce a good match to hand annotation of behavioural states. However, we find many behavioural events cannot be unambiguously classified. By comparing the results with ESA of an artificial agent''s behaviour, we argue that the ambiguity is due to greater continuity between behavioural states than is generally assumed for these organisms.     

Volatile organic compounds emitted by filamentous fungi isolated from flooded homes after Hurricane Sandy show toxicity in a Drosophila bioassay

Superstorm Sandy provided an opportunity to study filamentous fungi (molds) associated with winter storm damage. We collected 36 morphologically distinct fungal isolates from flooded buildings. By combining traditional morphological and cultural characters with an analysis of ITS sequences (the fungal DNA barcode), we identified 24 fungal species that belong to eight genera: Penicillium (11 species), Fusarium (four species), Aspergillus (three species), Trichoderma (two species), and one species each of Metarhizium, Mucor, Pestalotiopsis, and Umbelopsis. Then, we used a Drosophila larval assay to assess possible toxicity of volatile organic compounds (VOCs) emitted by these molds. When cultured in a shared atmosphere with growing cultures of molds isolated after Hurricane Sandy, larval toxicity ranged from 15 to 80%. VOCs from Aspergillus niger 129B were the most toxic yielding 80% mortality to Drosophila after 12 days. The VOCs from Trichoderma longibrachiatum 117, Mucor racemosus 138a, and Metarhizium anisopliae 124 were relatively non‐toxigenic. A preliminary analysis of VOCs was conducted using solid‐phase microextraction‐gas chromatography‐mass spectrometry from two of the most toxic, two of the least toxic, and two species of intermediate toxicity. The more toxic molds produced higher concentrations of 1‐octen‐3‐ol, 3‐octanone, 3‐octanol, 2‐octen‐1‐ol, and 2‐nonanone; while the less toxic molds produced more 3‐methyl‐1‐butanol and 2‐methyl‐1‐propanol, or an overall lower amount of volatiles. Our data support the hypothesis that at certain concentrations, some VOCs emitted by indoor molds are toxigenic.     

Statistical analysis and modeling of the temperature-dependent sleep behavior of drosophila

The sleep behavior of drosophila is analyzed under different temperatures. The activity per minute of the flies is recorded automatically. Sleep for a fruit fly is defined as the periods without any activity and longer than 5 minutes. Several parameters such as total sleep time, circadian sleep profile, quality of sleep are analyzed. The sleep behaviors are significantly different for flies at different temperature. Interestingly, the durations of daytime sleep periods show a common scale-free power law distribution. We propose a stochastic model to simulate the activities of the population of neurons which regulate the dynamics of sleep–wake process to explain the distribution of daytime sleep.     

杏仁种皮黑色素对果蝇的紫外辐射保护作用

以果蝇西农12和果蝇中农e为实验模型,对果蝇进行紫外辐射处理后,在添加杏仁种皮黑色素的培养基中进行培养,通过测定果蝇繁殖力、寿命、超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量、蛋白质量浓度、总抗氧化能力(T-AOC)等指标来研究杏仁种皮黑色素的抗紫外辐射活性。结果表明:紫外辐射组与对照组相比,两品系果蝇的繁殖力、寿命、总抗氧化能力显著降低,蛋白质量浓度、超氧化物歧化酶活力呈现先增后降趋势,丙二醛含量显著升高。当添加杏仁种皮黑色素质量浓度范围为0.02～0.2mg/mL时,与2min UV组相比,能显著提高两品系果蝇的繁殖力,但所设添加黑色素的实验组中相间组的果蝇繁殖力没有达到显著水平;与5min UV组相比,添加黑色素能显著提高两品系果蝇的寿命、蛋白质量浓度、总抗氧化能力、SOD活力,降低MDA含量。杏仁种皮黑色素对果蝇具有抗紫外辐射保护作用。     

Managing the Public Sphere: Journalistic Construction of the Great Globalization Debate

There is little consensus on what constitutes open, deliberative media discourse. We offer a simple, measurable, and comparative model based on 3 aspects of source and issue construction in news accounts: access, recognition, and responsiveness. The model is applied to coverage of 2001–2003 World Economic Forum (WEF) meetings and protests against the organization's role in global economic policies. Both demonstrators and WEF participants were granted news access, but WEF actors were recognized more formally and given greater input in news content, including ownership claims to many activist issue positions. Journalistic deference to the WEF communication agenda limited mutual responsiveness. The journalistic process systematically managed the debate about globalization on terms that favored elites over citizen-activists.