Effect of UVM induction on mutation fixation at non-pairing and mispairing DNA lesions

Mutation fixation at an ethenocytosine (εC) residue borne on transfected M13 single-stranded DNA is significantly enhanced in response to pretreatment of Escherichia coli cells with UV, alkylating agents or hydrogen peroxide, a phenomenon that we have called UVM for UV modulation of mutagenesis. The UVM response does not require the E. coli SOS or adaptive responses, and is observed in cells defective for oxyR , an oxidative DNA damage-responsive regulatory gene. UVM may represent either a novel DNA-repair phenomenon, or an unrecognized feature of DNA replication in damaged cells that affects a specific class of non-coding DNA lesions. To explore the range of DNA lesions subject to the UVM effect, we have examined mutation fixation at 3, N  ⁴-ethenocytosine and 1, N  ⁶-ethenoadenine, as well as at O⁶-methylguanine (O⁶mG). M13 viral single-stranded DNA constructs bearing a single mutagenic lesion at a specific site were transfected into cells pretreated with UV or 1-methyl-3-nitro-1-nitrosoguanidine (MNNG). Survival of transfected viral DNA was measured as transfection efficiency, and mutagenesis at the lesion site was analysed by a quantitative multiplex sequence analysis technology. The results suggest that the UVM effect modulates mutagenesis at the two etheno lesions, but does not appear to significantly affect mutagenesis at O⁶mG. Because the modulation of mutagenesis is observed in cells incapable of the SOS response, these data are consistent with the notion that UVM may represent a previously unrecognized DNA damage-inducible response that affects the fidelity of DNA replication at certain mutagenic lesions in Escherichia coli .     

HLA-DO is a negative modulator of HLA-DM-mediated MHC class II peptide loading

Background: Class II molecules of the major histocompatibility complex become loaded with antigenic peptides after dissociation of invariant chainderived peptides (CLIP) from the peptide-binding groove. The human leukocyte antigen (HLA)-DM is a prerequisite for this process, which takes place in specialised intracellular compartments. HLA-DM catalyses the peptide-exchange process, simultaneously functioning as a peptide ‘editor’, favouring the presentation of stably binding peptides. Recently, HLA-DO, an unconventional class II molecule, has been found associated with HLA-DM in B cells, yet its function has remained elusive.Results: The function of the HLA-DO complex was investigated by expression of both chains of the HLA-DO heterodimer (either alone or fused to green fluorescent protein) in human Mel JuSo cells. Expression of HLA-DO resulted in greatly enhanced surface expression of CLIP via HLA-DR3, the conversion of class II complexes to the SDS-unstable phenotype and reduced antigen presentation to T-cell clones. Analysis of peptides eluted from HLA-DR3 demonstrated that CLIP was the major peptide bound to class II in the HLA-DO transfectants. Peptide exchange assays in vitro revealed that HLA-DO functions directly at the level of class II peptide loading by inhibiting the catalytic action of HLA-DM.Conclusions: HLA-DO is a negative modulator of HLA-DM. By stably associating with HLA-DM, the catalytic action of HLA-DM on class II peptide loading is inhibited. HLA-DO thus affects the peptide repertoire that is eventually presented to the immune system by MHC class II molecules.     

RAPID GENETIC DIFFERENTIATION AND FOUNDER EFFECT IN COLONIZING POPULATIONS OF COMMON MYNAS (ACRIDOTHERES TRISTIS)

Populations of common mynas introduced to Australia, New Zealand, Fiji, Hawaii, and South Africa from India during the last century were compared genetically with the extant native population using isozyme electrophoresis of 39 presumptive loci. Average heterozygosity, mean number of alleles/locus, and the percentage of polymorphic loci are lower in the introduced populations, and the 18% loss of alleles involves only alleles that are rare in the native population. The native population is only weakly subdivided genetically (F_ST = 0.032) whereas the introduced populations are much more differentiated (F_ST = 0.123), and the mean genetic distance among them is significantly greater than among native samples. The reduction in mean number of alleles/locus and average heterozygosity is greatest in the South African population, consistent with a very small effective size in the founder population. In the introduced populations, random drift is implicated by the different subsets of polymorphic loci they possess, by their greater variance in allele frequencies, and by shifts either side of the native means. It is concluded that in the evolutionarily short period of 100–120 years, bottlenecks and random drift have promoted genetic shifts equal to those between different subspecies of birds.     

Effect of Plant Factors, Sugar Contents, and Control Methods on the Top Borer (Scirpophaga nivella F.) Infestation in Selected Varieties of Sugarcane

Sugarcane (Saccharum officinarum L.) varieties CP‐43/33 and L‐118 susceptible, BF‐162 and SPSG‐26 resistant while CP‐72/2086 as intermediate were screened for studies on top borer, Scirpophaga nivella Fabricus (Pyralidae: Lepidoptera). Morpho‐physio chemical plant factors as well as quality of sugar were examined to elucidate the relationship between pest infestations. The observations on leaf thickness 0.735* and moisture contents 0.771* showed positive and significant correlation with the pest infestation at tillering stage. Total minerals, manganese and copper contents did not show significant correlation with the pest infestation, whereas nitrogen, potassium, calcium, magnesium and ferrous contents manifested positive and significant correlation with the pest infestation. Phosphorous, carbohydrates, fats and zinc produced significant and adverse effect on the pest infestation at tillering stage. Zinc contents with contrasting behaviors appeared to be the most important character with co‐efficient value of 0.764 followed by ferrous with positive sign. The effect of borer infestation was significantly negative on pol (sucrose), Brix contents (total soluble solids), and CCS (commercial cane sugar). The coefficient of determination value was 0.821, obtained by computing fiber content, pol, Brix and CCS factors together for multivariate regression models. Application of trash mulching at the time of sowing proved to be the most effective treatment with minimum infestation (3.91%) of borer and maximum protection level (62.87%) followed by Furadan®, removal of dead hearts + spike thrust, hand collection of egg masses, and cutting of shoots at 15 days interval.     

TEMPERATURE REQUIREMENTS FOR THE DEVELOPMENT AND SURVIVAL OF RICE STEMBORERS IN LABORATORY CONDITIONS

Abstract  The effect of seven constant temperatures from 10 to 40°C (10, 15, 20, 25 30, 35 and 40°C) on the development of eggs, larvae and pupae of rice stemborers viz., Chilo polychrysa (Meyrick), C. suppressalis (Walker), C. partellus (Swinhoe), Scirpophaga incertulas (Walker), S. innotata (Walker) and Sesamia inferens (Walker) were studied. The mean developmental period among constant temperatures (in days) of egg, larva and pupa of six borers differed significantly ( P < 0.0001). The mean percent of development per day of egg, larva and pupa of all borers gradually increased with the increase of constant temperatures. The total developmental period was inversely decreased with the increase of constant temperatures. The lower threshold temperature was found between 10–15°Cand higher threshold temperature between 35–40°C, where no development took place. The mean developmental zero was 8.57±1.71, 7.70±1.01, 8.56±3.25, 10.19±2.19, 8.64±2.68 and 7.91±0.82 for egg, larva and pupa of above-mentioned borers respectively. The total thermal constant of egg, larva and pupa was 705.56, 725.32, 703.30, 556.59, 655.34 and 837.95 degree- days for C. polychrysa, C. suppressalis, C. partellus, S. incertulas, S. innotata and S. inferens respectively. The degree- days required for oviposition of female moths of the six borers was calculated as 99.06, 90.85, 99.29, 75.16, 92.25 and 80.41 respectively. The total degree- days required completing a generation was 804.62, 816.17, 802.59, 631.75, 648.84 and 918.36 respectively.     

Angelica polymorpha Maxim Induces Apoptosis of Human SH-SY5Y Neuroblastoma Cells by Regulating an Intrinsic Caspase Pathway

Angelica polymorpha Maxim root extract (APRE) is a popular herbal medicine used for treating stomachache, abdominal pain, stomach ulcers, and rheumatism; however the effect of APRE on cancer cells has not yet been explored. Here, we examined APRE cytotoxicity seen on target neuroblastoma cells (NB) using cell viability assays, DAPI visualization of fragmented DNA, and Western blotting analysis of candidate signaling pathways involved in proliferation and apoptosis. We demonstrated that APRE reduced cell viability in NB to a greater extent than in fibroblast cells. In addition, we found that APRE could inhibit the three classes of MAPK proteins and could also down-regulate the PI3K/AKT/GSK-3β activity all being relevant for proliferation and survival. APRE could also up-regulate Bax expression and down-regulate Bcl-2 and Mcl-1. With APRE treatment, depolarization of mitochondria membrane potential and activation of caspase-3 was demonstrated in the SH-SY5Y cells. We could not found increased activity of death receptor and caspase-8 as markers of the extrinsic apoptosis pathway for the APRE treated cells. In presence of a caspase-3 siRNA and a pan-caspase inhibitor, APRE could not reduce the viability of NB cells to a significant degree. So we predicted that with APRE, the intrinsic pathway was solely responsible for inducing apoptosis as we also showed that the non-caspase autophagy pathway or ER stress-ROS mediated pathways were not involved. These findings demonstrate that an intrinsic mitochondria-mediated apoptosis pathway mediates the apoptotic effects of APRE on SH-SY5Y cells, and that APRE shows promise as a novel agent for neuroblastoma therapy.     

Incorporating service and user information and latent features to predict QoS for selecting and recommending cloud service compositions

The rapid growth of published cloud services in the Internet makes the service selection and recommendation a challenging task for both users and service providers. In cloud environments, software re services collaborate with other complementary services to provide complete solutions to end users. The service selection is performed based on QoS requirements submitted by end users. Software providers alone cannot guarantee users’ QoS requirements. These requirements must be end-to-end, representing all collaborating services in a cloud solution. In this paper, we propose a prediction model to compute end-to-end QoS values for vertically composed services which are composed of three types of cloud services: software (SaaS), infrastructure (IaaS) and data (DaaS) services. These values can be used during the service selection and recommendation process. Our model exploits historical QoS values and cloud service and user information to predict unknown end-to-end QoS values of composite services. The experiments demonstrate that our proposed model outperforms other prediction models in terms of the prediction accuracy. We also study the impact of different parameters on the prediction results. In the experiments, we used real cloud services’ QoS data collected using our developed QoS monitoring and collecting system.     

Binding interactions of the peripheral stalk subunit isoforms from human V-ATPase

The mammalian peripheral stalk subunits of the vacuolar-type H⁺-ATPases (V-ATPases) possess several isoforms (C1, C2, E1, E2, G1, G2, G3, a1, a2, a3, and a4), which may play significant role in regulating ATPase assembly and disassembly in different tissues. To better understand the structure and function of V-ATPase, we expressed and purified several isoforms of the human V-ATPase peripheral stalk: E1G1, E1G2, E1G3, E2G1, E2G2, E2G3, C1, C2, H, a1_NT, and a2_NT. Here, we investigated and characterized the isoforms of the peripheral stalk region of human V-ATPase with respect to their affinity and kinetics in different combination. We found that different isoforms interacted in a similar manner with the isoforms of other subunits. The differences in binding affinities among isoforms were minor from our in vitro studies. However, such minor differences from the binding interaction among isoforms might provide valuable information for the future structural-functional studies of this holoenzyme.