Integrity and Function of the Saccharomyces cerevisiae Spindle Pole Body Depends on Connections Between the Membrane Proteins Ndc1, Rtn1, and Yop1

The nuclear envelope in Saccharomyces cerevisiae harbors two essential macromolecular protein assemblies: the nuclear pore complexes (NPCs) that enable nucleocytoplasmic transport, and the spindle pole bodies (SPBs) that mediate chromosome segregation. Previously, based on metazoan and budding yeast studies, we reported that reticulons and Yop1/DP1 play a role in the early steps of de novo NPC assembly. Here, we examined if Rtn1 and Yop1 are required for SPB function in S. cerevisiae. Electron microscopy of rtn1Δ yop1Δ cells revealed lobular abnormalities in SPB structure. Using an assay that monitors lateral expansion of the SPB central layer, we found that rtn1Δ yop1Δ SPBs had decreased connections to the NE compared to wild type, suggesting that SPBs are less stable in the NE. Furthermore, large budded rtn1Δ yop1Δ cells exhibited a high incidence of short mitotic spindles, which were frequently misoriented with respect to the mother–daughter axis. This correlated with cytoplasmic microtubule defects. We found that overexpression of the SPB insertion factors NDC1, MPS2, or BBP1 rescued the SPB defects observed in rtn1Δ yop1Δ cells. However, only overexpression of NDC1, which is also required for NPC biogenesis, rescued both the SPB and NPC associated defects. Rtn1 and Yop1 also physically interacted with Ndc1 and other NPC membrane proteins. We propose that NPC and SPB biogenesis are altered in cells lacking Rtn1 and Yop1 due to competition between these complexes for Ndc1, an essential common component of both NPCs and SPBs.     

ChiloKey,an interactive identification tool for the geophilomorph centipedes of Europe (Chilopoda,Geophilomorpha)

ChiloKey is a matrix-based, interactive key to all 179 species of Geophilomorpha (Chilopoda) recorded from Europe, including species of uncertain identity and those whose morphology is known partially only. The key is intended to assist in identification of subadult and adult specimens, by means of microscopy and simple dissection techniques whenever necessary. The key is freely available through the web at: http://www.biologia.unipd.it/chilokey/ and at http://www.interactive-keys.eu/chilokey/.     

Parasitoids of whiteflies (Hymenoptera: Aphelinidae,Eulophidae, Platygastridae; Hemiptera: Aleyrodidae) from the Macaronesian archipelagos of the Canary Islands,Madeira and the Azores

Data on the whitefly parasitoid species known from the Macaronesian archipelagos of the Canary Islands, Madeira and the Azores are presented, based largely on recently collected material. A total of 26 species are treated, including six new species, six new records for the Canary Islands, two new records for Madeira, and two new records for the Azores. All species are fully described and illustrated. New species described are: Encarsia atlantica Polaszek & Hernández; Encarsia levadicola Polaszek & Hernández; Encarsia melanostoma Polaszek & Hernández; Encarsia noahi Polaszek & Hernández; Euderomphale gomer LaSalle & Hernández; Euderomphale insularis LaSalle & Hernández. A fully illustrated identification key based on females is provided for recognition of whitefly parasitoids in these archipelagos. Data on the known distribution and hosts are provided, as well as references to biology and use in biological control.     

Allosteric Inhibitors Have Distinct Effects,but Also Common Modes of Action,in the HCV Polymerase

The RNA-dependent RNA polymerase from the Hepatitis C Virus (gene product NS5B) is a validated drug target because of its critical role in genome replication. There are at least four distinct allosteric sites on the polymerase to which several small molecule inhibitors bind. In addition, numerous crystal structures have been solved with different allosteric inhibitors bound to the polymerase. However, the molecular mechanisms by which these small molecules inhibit the enzyme have not been fully elucidated. There is evidence that allosteric inhibitors alter the intrinsic motions and distribution of conformations sampled by the enzyme. In this study we use molecular dynamics simulations to understand the structural and dynamic changes that result when inhibitors are bound at three different allosteric binding sites on the enzyme. We observe that ligand binding at each site alters the structure and dynamics of NS5B in a distinct manner. Nonetheless, our studies also highlight commonalities in the mechanisms of action of the different inhibitors. Each inhibitor alters the conformational states sampled by the enzyme, either by rigidifying the enzyme and preventing transitions between functional conformational states or by destabilizing the enzyme and preventing functionally relevant conformations from being adequately sampled. By illuminating the molecular mechanisms of allosteric inhibition, these studies delineate the intrinsic functional properties of the enzyme and pave the way for designing novel and more effective polymerase inhibitors. This information may also be important to understand how allosteric regulation occurs in related viral polymerases and other enzymes.     

Signaling by the Engulfment Receptor Draper: A Screen in Drosophila melanogaster Implicates Cytoskeletal Regulators,Jun N-Terminal Kinase,and Yorkie

Draper, the Drosophila melanogaster homolog of the Ced-1 protein of Caenorhabditis elegans, is a cell-surface receptor required for the recognition and engulfment of apoptotic cells, glial clearance of axon fragments and dendritic pruning, and salivary gland autophagy. To further elucidate mechanisms of Draper signaling, we screened chromosomal deficiencies to identify loci that dominantly modify the phenotype of overexpression of Draper isoform II (suppressed differentiation of the posterior crossvein in the wing). We found evidence for 43 genetic modifiers of Draper II. Twenty-four of the 37 suppressor loci and 3 of the 6 enhancer loci were identified. An additional 5 suppressors and 2 enhancers were identified among mutations in functionally related genes. These studies reveal positive contributions to Drpr signaling for the Jun N-terminal Kinase pathway, supported by genetic interactions with hemipterous, basket, jun, and puckered, and for cytoskeleton regulation as indicated by genetic interactions with rac1, rac2, RhoA, myoblast city, Wiskcott–Aldrich syndrome protein, and the formin CG32138, and for yorkie and expanded. These findings indicate that Jun N-terminal Kinase activation and cytoskeletal remodeling collaborate in Draper signaling. Relationships between Draper signaling and Decapentaplegic signaling, insulin signaling, Salvador/Warts/Hippo signaling, apical-basal cell polarity, and cellular responses to mechanical forces are also discussed.     

Inhibition of the sodium channel by SK&F 96365, an inhibitor of the receptor-operated calcium channel,in mouse diaphragm

The effects of SK&F 96365, a blocker of the receptor-operated Ca²⁺ channel, on contractilities and the Na⁺ channel of mouse diaphragm were studied. SK&F 96365 (10–50 µM) reversibly inhibited twitches, tetanic contractions and muscle and nerve action potentials. The IC₅₀ was 17–24 µM. The inward Na⁺ current was suppressed and its recovery from inactivations delayed. Crotamine, a peptide toxin that binds to neurotoxin receptor site 3 of the muscle Na⁺ channel, enhanced the inhibitory effects of SK&F 96365 and reduced the IC₅₀ to about 4 µM. Veratridine had similar effects, although it was less effective than crotamine. On the other hand, the crotamine-induced membrane depolarizations and spontaneous discharges of muscle action potentials were inhibited by SK&F 96365 noncompetitively. The inhibitory effects of tetrodotoxin and tetracaine were additive with those of SK&F 96365 but were enhanced slightly by crotamine. The results suggested that SK&F 96365 acts on a distinct site and blocks the Na⁺ channel of excitable membranes at a concentration range that inhibits the receptor-operated calcium channel.     

A cultivar comparison of plant regeneration from suspension cells,callus, and cormel slices ofGladiolus

Callus was initiated from either cormel slices or in vitro-grown plants of sixGladiolus cultivars cultured on Murashige and Skoog’s basal salts medium supplemented with either 10 mg/liter (53.8 µM) 1-naphthaleneacetic acid, 2 mg/liter (9.3 µM) dicamba, or 0.5 mg/liter (2.2 µM) 2,4-dichlorophenoxyacetic acid. More plants were regenerated from callus of the cultivar “Peter Pears” as compared to “Jenny Lee,” “Florida Flame,” or “Golden Year.” No plants were regenerated from callus of “Rosa Supreme” or “Purity White.” Plants were regenerated from 2 and 6-mo.-old suspension cells of “Jenny Lee” and “Peter Pears” but not from “Florida Flame.” Cormel slices cultured on Murashige and Skoog’s basal salts medium supplemented with 1 mg/liter (4.4 µM) 6-benzylaminopurine regenerated plants from all six cultivars indicating a cultivar-independent system of plant regeneration.     

Uptake of Home-Based HIV Testing,Linkage to Care,and Community Attitudes about ART in Rural KwaZulu-Natal,South Africa: Descriptive Results from the First Phase of the ANRS 12249 TasP Cluster-Randomised Trial

BackgroundThe 2015 WHO recommendation of antiretroviral therapy (ART) for all immediately following HIV diagnosis is partially based on the anticipated impact on HIV incidence in the surrounding population. We investigated this approach in a cluster-randomised trial in a high HIV prevalence setting in rural KwaZulu-Natal. We present findings from the first phase of the trial and report on uptake of home-based HIV testing, linkage to care, uptake of ART, and community attitudes about ART.ConclusionsHome-based HIV testing was well received in this rural population, although men were less easily contactable at home; immediate ART was acceptable, with good viral suppression and retention. However, only about half of HIV-positive people accessed care within 6 mo of being identified, with nearly two-thirds accessing care by 12 mo. The observed delay in linkage to care would limit the individual and public health ART benefits of universal testing and treatment in this population.

Trial registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01509508","term_id":"NCT01509508"}}NCT01509508     

Revision of the Thelastomatoidea,Oxyurida of invertebrate hosts III. Hystrignathidae

The family Hystrignathidae (Thelastomatoidea, Oxyurida, Nematoda) is revised. The following genera are included in the family:Anomalostoma Cordeira,Anuronema Clark,Artigasia Christie,Boraceianema Travasssos & Kloss,Carlosia Travassos & Kloss,Christiella Travassos & Kloss,Coronocephalus Cordeira,Glaber Travassos & Kloss,Hystrignathus Leidy,Klossiella Cordeira,Klossnema Cordeira,Lepidonema Cobb,Longior Travassos & Kloss,Mentecle Travassos & Kloss,Papillabrum Kloss,Passalidophila Van Waerebeke,Phalacronema Clark,Salesia Travassos & Kloss,Soaresnema Travassos & Kloss,Sprentia Clark,Triumphalisnema Kloss,Urbanonema Travassos & Kloss,Ventelia Travassos & Kloss,Vulcanonema Travassos & Kloss andXyo Cobb.Paraxyo Travassos and Kloss is considered a synonym ofArtigasia Cobb with the following resulting new combinations:A. ensicrinatus (Hunt) andA. monodelpho (Travassos & Kloss).Artigasia pauliani var.joliveta Théodoridès is raised to the rank of species. Complete diagnoses are given to all genera and annotated lists of species are provided.     

Birds do it,bees do it,worms and ciliates do it too: DNA methylation from unexpected corners of the tree of life

Studies describing intricate patterns of DNA methylation in nematode and ciliate are controversial due to the uncertainty of genomic evolutionary conservation of DNA methylation enzymes.See related research articles http://genomebiology.com/2012/13/10/R99 and http://genomebiology.com/2012/13/10/R100     

Coordination of Recombination with Meiotic Progression in the Caenorhabditis elegans Germline by KIN-18, a TAO Kinase That Regulates the Timing of MPK-1 Signaling

Meiosis is a tightly regulated process requiring coordination of diverse events. A conserved ERK/MAPK-signaling cascade plays an essential role in the regulation of meiotic progression. The Thousand And One kinase (TAO) kinase is a MAPK kinase kinase, the meiotic role of which is unknown. We have analyzed the meiotic functions of KIN-18, the homolog of mammalian TAO kinases, in Caenorhabditis elegans. We found that KIN-18 is essential for normal meiotic progression; mutants exhibit accelerated meiotic recombination as detected both by analysis of recombination intermediates and by crossover outcome. In addition, ectopic germ-cell differentiation and enhanced levels of apoptosis were observed in kin-18 mutants. These defects correlate with ectopic activation of MPK-1 that includes premature, missing, and reoccurring MPK-1 activation. Late progression defects in kin-18 mutants are suppressed by inhibiting an upstream activator of MPK-1 signaling, KSR-2. However, the acceleration of recombination events observed in kin-18 mutants is largely MPK-1-independent. Our data suggest that KIN-18 coordinates meiotic progression by modulating the timing of MPK-1 activation and the progression of recombination events. The regulation of the timing of MPK-1 activation ensures the proper timing of apoptosis and is required for the formation of functional oocytes. Meiosis is a conserved process; thus, revealing that KIN-18 is a novel regulator of meiotic progression in C. elegans would help to elucidate TAO kinase’s role in germline development in higher eukaryotes.     

The taxonomy of the family Paramphistomidae Fischoeder, 1901 with special reference to the morphology of species occurring in ruminants. V. Revision of the genus Cotylophoron Stiles & Goldberger, 1910

Summary Only seven species of the genus Cotylophoron Stiles & Goldberger, 1910 are considered valid, namely C. cotylophorum (Fischoeder, 1901) Stiles & Goldberger, 1910 (type species); C. jacksoni N?smark, 1937; C. fuelleborni N?smark, 1937; C. panamensis Price and McIntosh, 1953; C. bareilliensis Mukherjee & Chauhan, 1965; C. macrosphinctris Sey & Graber, 1979 and C. xiangjiangense Wang, 1979. These are redescribed and illustrated. Scanning electron photomicrographs of the tegumental surfaces of some species are provided. C. guangdongense Wang, 1979 is considered a synonym of C. cotylophorum (Fischoeder, 1901) Stiles & Goldberger 1910, and C. indicum of N?smark, 1937 (not of Stiles & Goldberger, 1910) and C. noveboracensis Price & McIntosh, 1953 are considered synonyms of C. fuelleborni N?smark, 1937. C. ottoi Gupta & Bakhshi in Gupta & Nakhasi, 1977 is regarded as a species inquirenda. A key to the species of the genus is given. Part of a thesis approved by the University of London for the award of the Ph.D degree.     

Pleiotrophin,A Multifunctional Tumor Promoter Through Induction of Tumor Angiogenesis,Remodeling of the Tumor Microenvironment,and Activation of Stromal Fibroblasts

Pleiotrophin (PTN, Ptn) is a widely expressed, developmentally regulated 136 amino acid secreted heparin-binding cytokine. It signals through a unique signaling pathway; the PTN receptor is the transmembrane receptor protein tyrosine phosphatase (RPTP)β/ζ. RPTPβ/ζ is inactivated by PTN, which leads to increased tyrosine phosphorylation of the downstream targets of the PTN/RPTPβ/ζ signaling pathway. Pleiotrophin gene expression is found in cells in early differentiation during different developmental periods. It is upregulated in cells with an early differentiation phenotype in wound repair. The Ptn gene also is a proto-oncogene; PTN is expressed in human tumor cells, and, in cell lines derived from human tumors that express Ptn, Ptn expression is constitutive and thus "inappropriate". Importantly, properties of different cells induced by PTN in PTN-stimulated cells are strikingly similar to properties of highly malignant cells. Furthermore, transformed cells into which Ptn is introduced undergo "switches" to malignant cells of higher malignancy with properties that are strikingly similar to properties of PTN-stimulated cells. These unique features of PTN support the conclusion that constitutive PTN signaling in malignant cells that inappropriately express Ptn functions as a potent tumor promoter. Recently, in confirmation, Ptn targeted by the mouse mammary tumor virus (MMTV) promoter in a transgenic mouse model was found to promote breast cancers to a more aggressive breast cancer cell phenotype that morphologically closely resembles scirrhous carcinoma in human; in addition, it promoted a striking increase in tumor angiogenesis and a remarkable degree of remodeling of the micro-environment. Pleiotrophin thus regulates both different normal and pathological functions; collectively, the different studies have uncovered the unique ability of a single cytokine PTN, which signals through the unique PTN/RPTPβ/ζ signaling pathway, to induce the many properties associated with tumor promotion in the malignant cells that constitutively express Ptn and in their microenvironment.     

Sustainability criteria and indicators for the Makiling Botanic Gardens in Los Baños,Laguna, Philippines: An application of the CIFOR C&I toolbox

The Center for International Forestry Research (CIFOR) generic template was used as an initial set of sustainability criteria and indicators (C&I) in the development of a unique C&I set for the Makiling Botanic Gardens (MBG). The MBG is a forest area managed for educational and recreational activities inside the University of the Philippines Los Baños campus. A multi-disciplinary team evaluated each criterion and indicator of the CIFOR generic template using multi-criteria analysis (MCA) techniques such as ranking, rating and pairwise comparison to eliminate irrelevant and inapplicable C&I items. The remaining C&I set was then restructured and additional items developed to finalize the set. The final C&I set for MBG was tested through a preliminary assessment conducted by MBG management and users. Results showed that the C&I set was appropriate for MBG, and focused primarily on the protection of ecological processes, while at the same time supporting recreational and educational activities. The MBG C&I also highlighted the importance of a sound policy framework that supports management, as well as the critical role that planning, implementation and monitoring play in progressing sustainability. Moreover, the study demonstrated the practical applicability of the CIFOR C&I Toolbox when developing C&I at the forest management unit level.     

Lake-wide assessment of microplastics in the surface waters of Lake Baikal,Siberia

Limnology - Small microplastic particles < 330 µm, sometimes called mini-microplastics (MMP), are far more abundant than those larger than 330 µm....