Microfilament stabilization by jasplakinolide arrests oocyte maturation, cortical granule exocytosis, sperm incorporation cone resorption, and cell-cycle progression, but not DNA replication, during fertilization in mice

Jasplakinolide (JAS), which induces microfilament polymerization and stabilization, inhibits microfilament-mediated events in murine oocyte maturation and fertilization in a fashion unlike the effects of cytochalasin B (CCB) and latranculin A (LAT A). JAS prevents egg polar body emission at a much lower concentration than either CCB or LAT A. Microfilament bundles were detected on the entire egg cortex after JAS exposure. Conversely, microfilament patterns did not change after exposure to CCB, and few microfilaments were observed after exposure to LAT A. Eggs that were allowed to recover from JAS were unable to recover normal microfilament organization. During oocyte maturation, JAS prevented both spindle migration to the oocyte cortex and first polar body emission. During in vitro fertilization, sperm head entered the eggs and formed pronuclei, but sperm tail entry, pronuclear centration, and second polar body emission were not detected. DNA synthesis occurs in these JAS-treated zygotes. JAS inhibited not only the formation, but also the disassembly, of incorporation cones. JAS was also found to prevent cortical granule exocytosis following artificial activation, and cortical granules were still beneath the plasma membrane even after activation. Finally, incorporation of microinjected nonmuscle actin into the microfilament network of mice eggs was delayed by JAS. We conclude that JAS acts as a microfilament inhibitor during maturation and fertilization and is more powerful than other inhibitors. Its mechanism differs in that it promotes assembly and stabilization of microfilaments. JAS is a novel cell permeable tool for the investigation of microfilament-dependent events in early mammalian development.     

Juvenile animal studies in the development of pediatric medicines: experience from European medicines and pediatric investigation plans

INTRODUCTION: The need for early consideration of pediatric investigation plans (PIP) to support an indication in pediatric population has led to an increased focus on the relevance of nonclinical studies in juvenile animals (JAS). The usefulness of JAS is not yet established and a criterion for request is still a learning process. OBJECTIVE: This article compares data from JAS in all medicines approved by European centralized procedure before Pediatric Regulation (1995–2005) and data from JAS in the nonclinical information on all approved PIP (2007–2009). RESULTS: Of the 226 substances licensed by centralized procedure in 10 years, 31.9% were considered for children and 31 JAS were described in 9.7%. Since 2007, of the 205 PIP decisions, 50 PIP (24.3%) have 87 JAS planned or requested. The mean number of JAS in each medicine or PIP, increased from 1.4 to 1.7 between the two periods and the juvenile rat remained as the prevalent species. CONCLUSIONS: Results demonstrate that JAS planned/performed in EU environment has significantly increased. Birth Defects Res (Part B) 92:353–358, 2011. © 2011 Wiley‐Liss, Inc.     

Mycoremediation of Endosulfan and Its Metabolites in Aqueous Medium and Soil by Botryosphaeria laricina JAS6 and Aspergillus tamarii JAS9

Microbial degradation offers an efficient and ecofriendly approach to remove toxicants from the contaminated environments. Botryosphaeria laricina JAS6 and Aspergillus tamarii JAS9 were capable of degrading endosulfan and their metabolites which were isolated through enrichment technique. Both the strains were able to withstand an exposure of 1300 mg/L and showed luxuriant growth at 1000 mg/L of endosulfan. The change in pH in the culture broth was from 6.8 to 3.4 and 3.8 during growth kinetic studies of JAS6 and JAS9 strains, respectively upon biological degradation of endosulfan. The degradation of endosulfan by JAS6 and JAS9 strains were examined by HPLC. The biodegradation rate constant (k) and the initial concentration were reduced by 50% (DT₅₀) which was determined by first and pseudo first order kinetic models. In the present investigation it has been revealed that Botryosphaeria laricina JAS6 and Aspergillus tamarii JAS9 possessing endosulfan degrading capability are being reported for the first time. These findings confirm the degradation of endosulfan by JAS6 and JAS9 strains which were accompanied by significant reduction in the toxicity and could be used as remedial measure in contaminated environments.     

The anti-proliferative agent jasplakinolide rearranges the actin cytoskeleton of plant cells.

In the present study, we have characterized the action of the natural cyclodepsipeptide jasplakinolide (JAS) on the cytoplasmic architecture, actin-based cytoplasmic motility, and the organization of the actin cytoskeleton in selected examples of green algae (Acetabularia, Pseudobryopsis and Nitella) and higher plant cells (Allium bulb scale cells and Sinapis root hairs). JAS was capable of influencing the actin cytoskeleton and inhibiting cytoplasmic streaming in a differential, cell type-specific manner. With the exception of Nitella, two consecutive responses were observed upon incubation with 2.5 microM JAS: In the first phase cytoplasmic streaming increased transiently alongside with minor modifications of the actin cytoskeleton in the form of adventitious actin spots and spikes appearing throughout the cell cortex in addition to the normal actin bundle system typical for each cell type. In the second phase, cytoplasmic streaming stopped and the actin cytoskeleton became heavily reorganized into shorter, straight, more and more randomly oriented bundle segments. JAS exerted severe long-term effects on the actin cytoskeleton when treatments exceeded 30min at a concentration of 2.5 microM. An in situ competition assay using equimolar concentrations of JAS and FITC-phalloidin suggested that JAS has a phalloidin-like action. Effects of JAS were significantly different from those of cytochalasin D with respect to the resulting degree of perturbance of cytoplasmic organization, the distribution of actin filaments and the speed of reversibility.     

Actin filament polymerization regulates gliding motility by apicomplexan parasites

Host cell entry by Toxoplasma gondii depends critically on actin filaments in the parasite, yet paradoxically, its actin is almost exclusively monomeric. In contrast to the absence of stable filaments in conventional samples, rapid-freeze electron microscopy revealed that actin filaments were formed beneath the plasma membrane of gliding parasites. To investigate the role of actin filaments in motility, we treated parasites with the filament-stabilizing drug jasplakinolide (JAS) and monitored the distribution of actin in live and fixed cells using yellow fluorescent protein (YFP)-actin. JAS treatment caused YFP-actin to redistribute to the apical and posterior ends, where filaments formed a spiral pattern subtending the plasma membrane. Although previous studies have suggested that JAS induces rigor, videomicroscopy demonstrated that JAS treatment increased the rate of parasite gliding by approximately threefold, indicating that filaments are rate limiting for motility. However, JAS also frequently reversed the normal direction of motility, disrupting forward migration and cell entry. Consistent with this alteration, subcortical filaments in JAS-treated parasites occurred in tangled plaques as opposed to the straight, roughly parallel orientation observed in control cells. These studies reveal that precisely controlled polymerization of actin filaments imparts the correct timing, duration, and directionality of gliding motility in the Apicomplexa.     

On the interaction of turbulent flow and feeding behaviour of juvenile Atlantic Salmon (Salmo salar)

Anthropogenic activities can dramatically modify the riverine habitat of juvenile Atlantic salmon ( Salmo salar ). In the perspective of protecting and restoring the fluvial habitat, bioenergetic models are often used to estimate fish habitat quality. These models determine the habitat quality as the ratio between the energetic gains (food) and costs (growth, metabolism) of a fish. The energetic costs of swimming in a river are generally estimated using the average flow velocity without consideration of the effect of turbulence. Juvenile Atlantic salmon (JAS) live in rivers characterized by intense velocity fluctuations, which are often described as a succession of high‐ and low‐speed flow regions. These flow structures are likely to affect the JAS activity that consists of long periods of sit‐and‐wait at the top of a protuberant rock interrupted by short bursting motions to capture drifting food particles. To minimize the energetic costs, it is hypothesized that JAS use low‐speed flow regions to initiate and undertake their feeding motions.  To improve bioenergetic modelling, this study aimed at analyzing the relation between turbulent flow structures and the feeding behaviour of JAS in a natural gravel‐bed river. We filmed eight JAS during 30 min with a submersible video camera while simultaneously measuring velocity fluctuations close to the fish in the St. Marguerite River, Quebec, Canada. Our results show that the proportion of time used for feeding motions decreases with increasing turbulent intensity and mean flow velocity; and that JAS do not seem to prefer low‐speed flow regions to initiate their feeding motions.     

Fluid-Phase Markers in the Basolateral Endocytic Pathway Accumulate in Response to the Actin Assembly-promoting Drug Jasplakinolide

To investigate the role of filamentous actin in the endocytic pathway, we used the cell-permeant drug Jasplakinolide (JAS) to polymerize actin in intact polarized Madin–Darby canine kidney (MDCK) cells. The uptake and accumulation of the fluid-phase markers fluorescein isothiocyanate (FITC)-dextran and horseradish peroxidase (HRP) were followed in JAS-treated or untreated cells with confocal fluorescence microscopy, biochemical assays, and electron microscopy. Pretreatment with JAS increased the uptake and accumulation of fluid-phase markers in MDCK cells. JAS increased endocytosis in a polarized manner, with a marked effect on fluid-phase uptake from the basolateral surface but not from the apical surface of polarized MDCK cells. The early uptake of FITC-dextran and HRP was increased more than twofold in JAS-treated cells. At later times, FITC-dextran and HRP accumulated in clustered endosomes in the basal and middle regions of JAS-treated cells. The large accumulated endosomes were similar to late endosomes but they were not colabeled for other late endosome markers, such as rab7 or mannose-6-phosphate receptor. JAS altered transport in the endocytic pathway at a later stage than the microtubule-dependent step affected by nocodazole. JAS also had a notable effect on cell morphology, inducing membrane bunching at the apical pole of MDCK cells. Although other studies have implicated actin in endocytosis at the apical cell surface, our results provide novel evidence that filamentous actin is also involved in the endocytosis of fluid-phase markers from the basolateral membrane of polarized cells.     

Biodegradation of chlorpyrifos and its hydrolyzing metabolite 3,5,6-trichloro-2-pyridinol by Sphingobacterium sp. JAS3

Biodegradation of chlorpyrifos and its metabolite 3,5,6-trichloro-2-pyridinol (TCP) were studied in aqueous medium and in soil with a novel bacterial strain JAS3. The molecular characterization based on 16S rRNA sequence analysis revealed the strain JAS3 as Sphingobacterium sp. The strain JAS3 was able to grow in minimal salt medium (MSM) supplemented with 300 mg l^?1 of chlorpyrifos as sole carbon source. The degradation of chlorpyrifos and its primary metabolite TCP were examined by HPLC. After 5 d, Sphingobacterium sp. JAS3 degraded chlorpyrifos and its metabolite TCP to benzene, 1,3-bis(1,1-dimethylethyl) was analyzed by GCMS. Degradation of chlorpyrifos and TCP in soil with and without addition of nutrients was also studied. The ability to degrade chlorpyrifos makes this strain a useful candidate for remediation of pesticide contaminated sites.     

混合糖发酵重组酿酒酵母的菌株构建和菊芋秸秆同步糖化发酵研究

【目的】构建可用于纤维素乙醇高效生产的混合糖发酵重组酿酒酵母菌株,并利用菊芋秸秆为原料进行乙醇发酵。【方法】筛选在木糖中生长较好的酿酒酵母YB-2625作为宿主菌,构建木糖共代谢菌株YB-2625 CCX。进一步通过r DNA位点多拷贝整合的方式,以YB-2625 CCX为出发菌株构建木糖脱氢酶过表达菌株,并筛选得到优势菌株YB-73。采用同步糖化发酵策略研究YB-73的菊芋秸秆发酵性能。【结果】YB-73菌株以90 g/L葡萄糖和30 g/L木糖为碳源进行混合糖发酵,乙醇产量比出发菌株YB-2625 CCX提高了13.9%,副产物木糖醇产率由0.89 g/g降低至0.31 g/g,下降了64.6%。利用重组菌YB-73对菊芋秸秆进行同步糖化发酵,48 h最高乙醇浓度达到6.10%(体积比)。【结论】通过转入木糖代谢途径以及r DNA位点多拷贝整合过表达木糖脱氢酶基因可有效提高菌株木糖发酵性能,并用于菊芋秸秆的纤维素乙醇生产。这是首次报道利用重组酿酒酵母进行菊芋秸秆原料的纤维素乙醇发酵。     

Biomolecular Structure and Composition Changes in the Root System of Long-Lipped Serapias (Serapias vomeracea) After In Vitro Chitosan and Plant Growth Regulator Treatments

In the current study, effects of chitosan oligomers (CHI-OM) with different polymerization degrees (DP) between 2 and 15, and polymeric chitosan (CHI-P) with a DP of 70 were compared with kinetin (KIN), 6-benzylaminopurine (BAP), indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), and jasmonic acid (JAS) on the structure and composition of several biomolecules in the root system of Serapias vomeracea. Evaluation of molecular alterations in the root system of S. vomeracea through an infrared spectroscopic approach provided insight into the differentiation between action mechanisms of chitosan and commonly used plant growth regulators. The results revealed that CHI-P and JAS treatments at low concentrations might enhance the lignin content in the cell walls. Also, JAS, CHI-OM, and CHI-P treatments enhanced cell wall lignification. The water-associated cellulose content of the cell walls was affected by the DP of chitosan. Membrane lipid stabilization and protein content were enhanced after CHI-P, JAS, and KIN treatments, while BAP and CHI-OM treatments triggered lipid, protein, and cell wall polysaccharide synthesis. In particular, CHI-OM treatment also enhanced rhamnogalacturonan and β-galactan content as well as xyloglucans and glucomannans, while the auxin treatments had a similar impact only on glucomannan content. These findings suggest that chitosan may show either purine-based cytokinin-like or lipid-based jasmonic acid-like activity in the root system of S. vomeracea. Additionally, depending on its degree of polymerization, the effects of chitosan may vary on root system development in S. vomeracea.

     

The Relative Contributions of Facial Shape and Surface Information to Perceptions of Attractiveness and Dominance

Although many studies have investigated the facial characteristics that influence perceptions of others’ attractiveness and dominance, the majority of these studies have focused on either the effects of shape information or surface information alone. Consequently, the relative contributions of facial shape and surface characteristics to attractiveness and dominance perceptions are unclear. To address this issue, we investigated the relationships between ratings of original versions of faces and ratings of versions in which either surface information had been standardized (i.e., shape-only versions) or shape information had been standardized (i.e., surface-only versions). For attractiveness and dominance judgments of both male and female faces, ratings of shape-only and surface-only versions independently predicted ratings of the original versions of faces. The correlations between ratings of original and shape-only versions and between ratings of original and surface-only versions differed only in two instances. For male attractiveness, ratings of original versions were more strongly related to ratings of surface-only than shape-only versions, suggesting that surface information is particularly important for men’s facial attractiveness. The opposite was true for female physical dominance, suggesting that shape information is particularly important for women’s facial physical dominance. In summary, our results indicate that both facial shape and surface information contribute to judgments of others’ attractiveness and dominance, suggesting that it may be important to consider both sources of information in research on these topics.     

Performance Evaluation of TCP over Optical Channels and Heterogeneous Networks

Next generation optical networks will soon provide users the capability to request and obtain end-to-end all optical 10 Gbps channels on demand. Individual users will use these channels to exchange large amounts of data and support applications for scientific collaborative work. These new applications, which expect steady transfer rates in the order of Gbps, will very likely use either TCP or a new transport layer protocol as the end-to-end communication protocol. In this paper, we investigate the performance of TCP and newer TCP versions over High Bandwidth Delay Product Channels (HBDPC), such as the on demand optical channels described above. In addition, we investigate the performance of these new TCP versions over wireless networks and according to old issues such as fairness. This is particularly important to make adoption decisions. Using simulations, we show that (1) the window-based mechanism of current TCP implementations is not suitable to achieve high link utilization and (2) congestion control mechanisms, such as the one utilized by TCP Vegas and Westwood are more appropriate and provide better performance. We also show that new TCP proposals, although perform better than current TCP versions, they still perform worse than TCP Vegas. In addition, we found that even though these newer versions improve TCP's performance over their original counterparts in HBDPC, they still have performance problems in wireless networks and present worse fairness problems than their old counterparts. We conclude that all these versions are still based on TCP's AIMD strategy or similar and therefore continue to be fairly blind in the way they increase and decrease their transmission rates. TCP will not be able to utilize the foreseen optical infrastructure adequately and support future applications if not redesigned to scale.     

Psychometric analysis of behavior pattern A in the Spanish population and its relationship with personality variables

As a first step in the epidemiological study for CHD prevention among the Spanish population, the authors have analyzed two of the instruments for the assessment of the behavior pattern A (JAS and Bortner questionnaires). From a student sample (n = 302), a factor analysis of items and scales was made, relating pattern A scores to Eysenck's personality variables (EPQ). The importance of an accurate psychometric study of pattern A was emphasized, in order to select A subjects as specifically as possible.     

Human circovirus TT virus genotype 6 expresses six proteins following transfection of a full-length clone

The expression profile of the circovirus TTV has not yet been fully characterized. In this paper, we show that following transfection of a full-length viral clone of TTV genotype 6, each of the three virally encoded mRNAs is translated from two initiating AUGs, and therefore, the TTV genome generates at least six proteins. Localization studies of hemagglutinin-tagged versions of these proteins in fixed cells, and green fluorescent protein-tagged versions of these proteins in living cells, expressed following transfection, demonstrated that two were primarily nuclear, two were primarily cytoplasmic, and two were found throughout the cell.     

Beliefs and fears underlying type A behavior: what makes Sammy run--so fast and aggressively?

Price has proposed that Type A behavior allows individuals to cope with fears and anxieties engendered by beliefs they develop about their environment through socialization. Three investigations tested her cognitive social learning model of Type A behavior by developing measures of beliefs and fears, and validating them against a standardized Type A measure, the Jenkins Activity Survey (two studies), and measures of Time Urgency, Hostility, Time for Reverie and Self-Centeredness (one study). Respondents provided data by completing questionnaires. Three beliefs and four fears were assessed. Findings indicated that the measures of beliefs and fears had desirable psychometric properties and produced modest relationships with the JAS, Time Urgency and Hostility. Future research directions and practical implications are offered.     

UniSave: the UniProtKB sequence/annotation version database

SUMMARY: The UniProtKB Sequence/Annotation Version database (UniSave) is a comprehensive archive of UniProtKB/Swiss-Prot and UniProtKB/TrEMBL entry versions. All changed Swiss-Prot and TrEMBL entries are loaded into the UniSave as part of the public bi-weekly UniProtKB releases. Unlike the UniProtKB, which contains only the latest Swiss-Prot and TrEMBL entry versions, the UniSave provides access to previous versions of these entries. AVAILABILITY: http://www.ebi.ac.uk/uniprot/unisave     

A Changing Number of Alternative States in the Boreal Biome: Reproducibility Risks of Replacing Remote Sensing Products

Publicly available remote sensing products have boosted science in many ways. The openness of these data sources suggests high reproducibility. However, as we show here, results may be specific to versions of the data products that can become unavailable as new versions are posted. We focus on remotely-sensed tree cover. Recent studies have used this public resource to detect multi-modality in tree cover in the tropical and boreal biomes. Such patterns suggest alternative stable states separated by critical tipping points. This has important implications for the potential response of these ecosystems to global climate change. For the boreal region, four distinct ecosystem states (i.e., treeless, sparse and dense woodland, and boreal forest) were previously identified by using the Collection 3 data of MODIS Vegetation Continuous Fields (VCF). Since then, the MODIS VCF product has been updated to Collection 5; and a Landsat VCF product of global tree cover at a fine spatial resolution of 30 meters has been developed. Here we compare these different remote-sensing products of tree cover to show that identification of alternative stable states in the boreal biome partly depends on the data source used. The updated MODIS data and the newer Landsat data consistently demonstrate three distinct modes around similar tree-cover values. Our analysis suggests that the boreal region has three modes: one sparsely vegetated state (treeless), one distinct ‘savanna-like’ state and one forest state, which could be alternative stable states. Our analysis illustrates that qualitative outcomes of studies may change fundamentally as new versions of remote sensing products are used. Scientific reproducibility thus requires that old versions remain publicly available.     

Dye structure affects Taq DNA polymerase terminator selectivity

All DNA sequencing methods have benefited from the development of new F667Y versions of Taq DNA polymerase. However, terminator chemistry methods show less uniform peak height patterns when compared to primer chemistry profiles suggesting that the dyes and/or their linker arms affect enzyme selectivity. We have measured elementary nucleotide rate and binding constants for representative rhodamine- and fluorescein-labeled terminators to determine how they interact with F667 versions of Taq Pol I. We have also developed a rapid gel-based selectivity assay that can be used to screen and to quantify dye-enzyme interactions with F667Y versions of the enzyme. Our results show that 6-TAMRA-ddTTP behaves like unlabeled ddTTP, while 6-FAM-ddTTP shows a 40-fold reduction in the rate constant for polymerization without affecting ground-state nucleotide binding. Detailed mechanism studies indicate that both isomers of different fluorescein dyes interfere with a conformational change step which the polymerase undergoes following nucleotide binding but only when these dyes are attached to pyrimidines. When these same dyes are attached to purines by the same propargylamino linker arm, they show no effect on enzyme selectivity. These studies suggest that it may be possible to develop fluorescein terminators for thermocycle DNA sequencing methods for polymerases that do not discriminate between deoxy- and dideoxynucleotides.     

Afterword: Dark Anthropology in Papua New Guinea?

Ranging from colonial modernism to postcolonial disappointment, the papers in this collection explore the possibilities of Dark Anthropology and an Anthropology of the Good in Papua New Guinea. With these two prospects in mind, I consider what these papers tell us about the situations of rural people on the peripheries of large resource projects and those in ‘Last Places’ bypassed by development and the State. In all of these cases, difficult predicaments entail hardship or suffering, but are also met with responses seeking to realise varying versions of the good. This, in turn, prompts further questions about which and whose good are at issue amid a plurality of values. I conclude by suggesting that the ensemble of papers offers a retrospective on local versions of modernity as possibility contends with experience.