Genome-wide deficiency screen for the genomic regions responsible for heat resistance in Drosophila melanogaster

Background: Temperature adaptation is one of the most important determinants of distribution and population size of organisms in nature. Recently, quantitative trait loci (QTL) mapping and gene expression profiling approaches have been used for detecting candidate genes for heat resistance. However, the resolution of QTL mapping is not high enough to examine the individual effects of various genes in each QTL. Heat stress-responsive genes, characterized by gene expression profiling studies, are not necessarily responsible for heat resistance. Some of these genes may be regulated in association with the heat stress response of other genes. Results: To evaluate which heat-responsive genes are potential candidates for heat resistance with higher resolution than previous QTL mapping studies, we performed genome-wide deficiency screen for QTL for heat resistance. We screened 439 isogenic deficiency strains from the DrosDel project, covering 65.6% of the Drosophila melanogaster genome in order to map QTL for thermal resistance. As a result, we found 19 QTL for heat resistance, including 3 novel QTL outside the QTL found in previous studies. Conclusion: The QTL found in this study encompassed 19 heat-responsive genes found in the previous gene expression profiling studies, suggesting that they were strong candidates for heat resistance. This result provides new insights into the genetic architecture of heat resistance. It also emphasizes the advantages of genome-wide deficiency screen using isogenic deficiency libraries

Effects of small Hsp genes on developmental stability and microenvironmental canalization

Background: Progression of development has to be insulated from the damaging impacts of environmental and genetic perturbations to produce highly predictable phenotypes. Molecular chaperones, such as the heat shock proteins (HSPs), are known to buffer various environmental stresses, and are deeply involved in protein homeostasis. These characteristics of HSPs imply that they might affect developmental buffering and canalization. Results: We examined the role of nine Hsp genes using the GAL4/UAS-RNAi system on phenotypic variation of various morphological traits in Drosophila melanogaster. The stability of bristle number, wing size and wing shape was characterized through fluctuating asymmetry (FA) and the coefficient of variation (CV), or among-individual variation. Progeny of the GAL4/Hsp-RNAi crosses tended to have reduced trait means for both wing size and wing shape. Transcriptional knockdown of Hsp67Bc and Hsp22 significantly increased FA of bristle number, while knockdown of Hsp67Ba significantly increased FA and among-individual variation of wing shape but only in males. Suppression of Hsp67Bb expression significantly increased among-individual variation of bristle number. The knockdown of gene expression was confirmed for Hsp67Ba, Hsp67Bc, Hsp22, and Hsp67Bb. Correlation between FA and CV or among-individual variation of each trait is weak and not significant except for the case of male wing shape. Conclusion: Four small Hsp genes (Hsp22, Hsp67Ba, Hsp67Bb and Hsp67Bc) showed involvement in the processes of morphogenesis and developmental stability. Due to possible different functions in terms of developmental buffering of these small Hsps, phenotypic stability of an organism is probably maintained by multiple mechanisms triggered by different environmental and genetic stresses on different traits. This novel finding may lead to a better understanding of non-Hsp90 molecular mechanisms controlling variability in morphological traits

Induction of tumor-specific acquired immunity against already established tumors by selective stimulation of innate DEC-205+ dendritic cells

Two major distinct subsets of dendritic cells (DCs) are arranged to regulate our immune responses in vivo; 33D1+ and DEC-205+ DCs. Using anti-33D1-specific monoclonal antibody, 33D1+ DCs were successfully depleted from C57BL/6 mice. When 33D1+ DC-depleted mice were stimulated with LPS, serum IL-12, but not IL-10 secretion that may be mediated by the remaining DEC-205+ DCs was markedly enhanced, which may induce Th1 dominancy upon TLR signaling. The 33D1+ DC-depleted mice, implanted with syngeneic Hepa1-6 hepatoma or B16-F10 melanoma cells into the dermis, showed apparent inhibition of already established tumor growth in vivo when they were subcutaneously (sc) injected once or twice with LPS after tumor implantation. Moreover, the development of lung metastasis of B16-F10 melanoma cells injected intravenously was also suppressed when 33D1+ DC-deleted mice were stimulated twice with LPS in a similar manner, in which the actual cell number of NK1.1+CD3− NK cells in lung tissues was markedly increased. Furthermore, intraperitoneal (ip) administration of a very small amount of melphalan (l-phenylalanine mustard; l-PAM) (0.25 mg/kg) in LPS-stimulated 33D1+ DC-deleted mice helped to induce H-2Kb-restricted epitope-specific CD8+ cytotoxic T lymphocytes (CTLs) among tumor-infiltrating lymphocytes against already established syngeneic E.G7-OVA lymphoma. These findings indicate the importance and effectiveness of selective targeting of a specific subset of DCs, such as DEC-205+ DCs alone or with a very small amount of anticancer drugs to activate both CD8+ CTLs and NK effectors without externally added tumor antigen stimulation in vivo and provide a new direction for tumor immunotherapy

Log N - Log S Relations and Spectral Properties of Sources from the ASCA Large Sky Survey --- their Implications for the Origin of the Cosmic X-ray Background (CXB)

We carried out the first wide-area unbiased survey with the ASCA satellite in the 0.7-10 keV band around a north Galactic-pole region covering a continuous area of 7 square degrees (Large Sky Survey; LSS). To make the best use of ASCA capability, we developed a new source-detection method where the complicated detector responses are fully taken into account. Applying this method to the entire LSS data independently in the total (0.7-7 keV), hard (2-10 keV), and soft (0.7-2 keV) band, we detected 107 sources altogether with sensitivity limits of 6 x 10E-14 (0.7-7 keV), 1 x 10E-13 (2-10 keV), and 2 x 10E-14 erg sE-1 cmE-2 (0.7-2 keV), respectively. A complete list of the detected sources is presented. Based on detailed studies by Monte Carlo simulations, we evaluated effects of the source confusion and accurately derived Log N - Log S relation in each survey band. The Log N - Log S relation in the hard band is located on the extrapolation from the GINGA and HEAO1 results with the Euclidean slope of -3/2, while that in the soft band is consistent with the results by ROSAT. At these flux limits, 30 (+/- 3) percent of the CXB in the 0.7-7 keV band and 23 (+/- 3) percent in the 2-10 keV band have been resolved into discrete sources. The average spectrum of faint sources detected in the total band shows a photon index of 1.63 +/- 0.07 in the 0.7-10 keV range, consistent with the comparison of source counts between the hard and the soft energy band. Those detected in the hard band show a photon index of 1.49 +/- 0.10 in the 2-10 keV range. These spectral properties suggest that contribution of sources with hard energy spectra become significant at a flux of 10E-13 erg sE-1 cmE-2 (2-10 keV). The most plausible candidates are type-II AGNs, as indicated by on-going optical identifications.Comment: 28 pages, 11 figures, to appear in ApJ 518, 1999; figure 1 replaced, minor errors in text correcte

Discovery of the Central Excess Brightness in Hard X-rays in the Cluster of Galaxies Abell 1795

Using the X-ray data from \ASCA, spectral and spatial properties of the intra-cluster medium (ICM) of the cD cluster Abell 1795 are studied, up to a radial distance of $\sim 12'$ ($\sim 1.3$ $h_{50}^{-1}$ kpc). The ICM temperature and abundance are spatially rather constant, although the cool emission component is reconfirmed in the central region. The azimuthally- averaged radial X-ray surface brightness profiles are very similar between soft (0.7--3 keV) and hard (3--10 keV) energy bands, and neither can be fitted with a single-$\beta$ model due to a strong data excess within $\sim5'$ of the cluster center. In contrast, double-$\beta$ models can successfully reproduce the overall brightness profiles both in the soft and hard energy bands, as well as that derived with the \ROSAT PSPC. Properties of the central excess brightness are very similar over the 0.2--10 keV energy range spanned by \ROSAT and \ASCA. Thus, the excess X-ray emission from the core region of this cluster is confirmed for the first time in hard X-rays above 3 keV. This indicates that the shape of the gravitational potential becomes deeper than the King-type one towards the cluster center. Radial profiles of the total gravitating matter, calculated using the double-$\beta$ model, reveal an excess mass of $\sim 3 \times 10^{13}~ M_{\odot}$ within $\sim 150 h^{-1}_{50}$ kpc of the cluster center. This suggests a hierarchy in the gravitational potential corresponding to the cD galaxy and the entire cluster.Comment: 27 pages, 8 figures; to appear ApJ 500 (June 20, 1998

Seven-Signal Proteomic Signature for Detection of Operable Pancreatic Ductal Adenocarcinoma and Their Discrimination from Autoimmune Pancreatitis

There is urgent need for biomarkers that provide early detection of pancreatic ductal adenocarcinoma (PDAC) as well as discrimination of autoimmune pancreatitis, as current clinical approaches are not suitably accurate for precise diagnosis. We used mass spectrometry to analyze protein profiles of more than 300 plasma specimens obtained from PDAC, noncancerous pancreatic diseases including autoimmune pancreatitis patients and healthy subjects. We obtained 1063 proteomic signals from 160 plasma samples in the training cohort. A proteomic signature consisting of 7 mass spectrometry signals was used for construction of a proteomic model for detection of PDAC patients. Using the test cohort, we confirmed that this proteomic model had discrimination power equal to that observed with the training cohort. The overall sensitivity and specificity for detection of cancer patients were 82.6% and 90.9%, respectively. Notably, 62.5% of the stage I and II cases were detected by our proteomic model. We also found that 100% of autoimmune pancreatitis patients were correctly assigned as noncancerous individuals. In the present paper, we developed a proteomic model that was shown able to detect early-stage PDAC patients. In addition, our model appeared capable of discriminating patients with autoimmune pancreatitis from those with PDAC

Genome-wide deficiency mapping of the regions responsible for temporal canalization of the developmental processes of Drosophila melanogaster

Developmental processes of organisms are programed to proceed in a finely regulated manner and finish within a certain period of time depending on the ambient environmental conditions. Therefore, variation in the developmental period under controlled genetic and environmental conditions indicates innate instability of the developmental process. In this study, we aimed to determine whether a molecular machinery exists that regulates the canalization of the developmental period and, if so, to test whether the same mechanism also stabilizes a morphological trait. To search for regions that influence the instability of the developmental period, we conducted genome-wide deficiency mapping with 441 isogenic deficiency strains covering 65.5% of the Drosophila melanogaster genome. We found that 11 independent deficiencies significantly increased the instability of the developmental period and 5 of these also significantly increased the fluctuating asymmetry of wing shape although there was no significant correlation between the instabilities of developmental period and wing shape in general. These results suggest that canalization processes of the developmental period and morphological traits are at least partially independent. Our findings emphasize the potential importance of temporal variation in development as an indicator of developmental stability and canalization and provide a novel perspective for understanding the regulation of phenotypic variability

Natural genetic variation in fluctuating asymmetry of wing shape in Drosophila melanogaster

Fluctuating asymmetry (FA), defined as random deviation from perfect symmetry, has been used to assay the inability of individuals to buffer their developmental processes from environmental perturbations (i.e., developmental instability). In this study, we aimed to characterize the natural genetic variation in FA of wing shape in Drosophila melanogaster, collected from across the Japanese archipelago. We quantified wing shapes at whole wing and partial wing component levels and evaluated their mean and FA. We also estimated the heritability of the mean and FA of these traits. We found significant natural genetic variation in all the mean wing traits and in FA of one of the partial wing components. Heritability estimates for mean wing shapes were significant in two and four out of five wing traits in males and females, respectively. On the contrary, heritability estimates for FA were low and not significant. This is a novel study of natural genetic variation in FA of wing shape. Our findings suggest that partial wing components behave as distinct units of selection for FA, and local adaptation of the mechanisms to stabilize developmental processes occur in nature