Development of zona-free hamster ova to blastocysts in vitro

The zona pellucida (ZP) enclosing the mammalian ovum is important for its protection and for initial stages of fertilization, but the role of the ZP during embryo development is less clear. This study was designed to investigate if the hamster ZP is needed for embryo development from 1-cell to blastocyst in vitro, and to compare methods for removing the ZP. A total of 395 hamster pronucleate ova were collected 10 h post activation from superovulated, mated female hamsters. The ZP was removed from some ova using either 0.05% pronase, 0.05% trypsin or acid Tyrode's solution. To prevent ZP-free ova from sticking together, they were cultured singly in 30-50 muL drops of HECM-6 culture medium together with ZP-intact ova as controls. There was no significant difference among treatment groups in embryo development to blastocyst: 36/87 (42%) in the ZP intact group; 35/75 (47%) in the pronase-treated ZP-free group; 37/74 (50%) in the trypsin-treated ZP-free group; and 37/71 (52%) in the acid-treated ZP-free group. These results indicate that 1) the ZP is unnecessary for hamster embryo development in vitro from the pronucleate ovum stage to blastocyst; 2) none of the three ZP-removal methods was detrimental to embryo development; 3) embryos do not need to be cultured in groups during in vitro development from 1-cell to blastocyst. (C) 2000 by Elsevier Science Inc.

Identification of a novel gene K23 over-expressed in fish cross-subfamily cloned embryos

A novel gene-K23, differentially expressed in cross-subfamily cloned embryos, was isolated by RACE-PCR technique. It had 2580 base pairs (bp) in length, with a 1,425 bp open reading frame (ORF) encoding a putative protein of 474 amino acids (aa). Bioinformatic analysis indicated that K23 had 22 phosphorylation sites, but it had no signal peptides. Developmental expression analysis in zebrafish showed that K23 transcripts were maternally expressed in ovum and the amount of K23 transcripts increased gradually from zygote to pharyngula period. Subcellular localization analysis revealed that K23 protein was homogeneously distributed both in nuclei and cytoplasm. Taken together, our findings indicate that K23 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos.

The precocious Chinese mitten crab: Changes of gonad, survival rate, and life span in a freshwater lake

Changes of the gonad. survival rate, and life span of the precocious (i.e., sexually mature in their first October-November, at the age of 5 or 6 months) Chinese mitten crab (Eriocheir sinensis) were studied in a small, shallow, macrophytic, freshwater lake along the middle reaches of the Changjiang River, China. The gonosomatic index (GSI) reached peak during the next March when the female GSI was 12.32 +/- 1.75 SD% and the male GSI A as 4.24 +/- 0.19%. The sexual glands degraded from then on. In the first ten-day period of the following July, there was no complete ovum in the ovary. and the sperm became thin and lost adhesion. The population declined sharply from November to July, and the last one (a female) died in the middle of July, which indicated that the life span of the precocious crab was about 12 months (from larval hatching in June to death in July of next year). The survival curve might be expressed as Y = 1.09exp(-0.018x) (Y survival rate; x: days) for the precocious crabs stocked in experimental cages.

兰州重离子加速器冷却储存环中束流位置测量系统的研制

介绍了兰州重离子加速器冷却储存环 (HIRFL CSRm)中束流位置测量系统设计的基本原理、PICKUP探针参数的选择以及有关物理量的计算。并完成了桌上实验 ,得出了有关结论。

Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts.

Many of the biochemical reactions of apoptotic cell death, including mitochondrial cytochrome c release and caspase activation, can be reconstituted in cell-free extracts derived from Xenopus eggs. In addition, because caspase activation does not occur until the egg extract has been incubated for several hours on the bench, upstream signaling processes occurring before full apoptosis are rendered accessible to biochemical manipulation. We reported previously that the adaptor protein Crk is required for apoptotic signaling in egg extracts (Evans, E.K., W. Lu, S.L. Strum, B.J. Mayer, and S. Kornbluth. 1997. EMBO (Eur. Mol. Biol. Organ.) J. 16:230-241). Moreover, we demonstrated that removal of Crk Src homology (SH)2 or SH3 interactors from the extracts prevented apoptosis. We now report the finding that the relevant Crk SH2-interacting protein, important for apoptotic signaling in the extract, is the well-known cell cycle regulator, Wee1. We have demonstrated a specific interaction between tyrosine-phosphorylated Wee1 and the Crk SH2 domain and have shown that recombinant Wee1 can restore apoptosis to an extract depleted of SH2 interactors. Moreover, exogenous Wee1 accelerated apoptosis in egg extracts, and this acceleration was largely dependent on the presence of endogenous Crk protein. As other Cdk inhibitors, such as roscovitine and Myt1, did not act like Wee1 to accelerate apoptosis, we propose that Wee1-Crk complexes signal in a novel apoptotic pathway, which may be unrelated to Wee1's role as a cell cycle regulator.

Control of Emi2 activity and stability through Mos-mediated recruitment of PP2A.

Before fertilization, vertebrate eggs are arrested in meiosis II by cytostatic factor (CSF), which holds the anaphase-promoting complex (APC) in an inactive state. It was recently reported that Mos, an integral component of CSF, acts in part by promoting the Rsk-mediated phosphorylation of the APC inhibitor Emi2/Erp1. We report here that Rsk phosphorylation of Emi2 promotes its interaction with the protein phosphatase PP2A. Emi2 residues adjacent to the Rsk phosphorylation site were important for PP2A binding. An Emi2 mutant that retained Rsk phosphorylation but lacked PP2A binding could not be modulated by Mos. PP2A bound to Emi2 acted on two distinct clusters of sites phosphorylated by Cdc2, one responsible for modulating its stability during CSF arrest and one that controls binding to the APC. These findings provide a molecular mechanism for Mos action in promoting CSF arrest and also define an unusual mechanism, whereby protein phosphorylation recruits a phosphatase for dephosphorylation of distinct sites phosphorylated by another kinase.

Long-duration animal tracking in difficult lighting conditions.

High-throughput analysis of animal behavior requires software to analyze videos. Such software typically depends on the experiments' being performed in good lighting conditions, but this ideal is difficult or impossible to achieve for certain classes of experiments. Here, we describe techniques that allow long-duration positional tracking in difficult lighting conditions with strong shadows or recurring "on"/"off" changes in lighting. The latter condition will likely become increasingly common, e.g., for Drosophila due to the advent of red-shifted channel rhodopsins. The techniques enabled tracking with good accuracy in three types of experiments with difficult lighting conditions in our lab. Our technique handling shadows relies on single-animal tracking and on shadows' and flies' being accurately distinguishable by distance to the center of the arena (or a similar geometric rule); the other techniques should be broadly applicable. We implemented the techniques as extensions of the widely-used tracking software Ctrax; however, they are relatively simple, not specific to Drosophila, and could be added to other trackers as well.

Ribonucleic acid metabolism in unfertilized and fertilized sea-urchin eggs.

Journal Article

Modulational instability of dust acoustic waves in dusty plasmas: Modulation obliqueness, background ion nonthermality, and dust charging effects

The oblique modulational instability of dust acoustic (DA) waves in an unmagnetized warm dusty plasma with nonthermal ions, taking into account dust grain charge variation (charging), is investigated. A nonlinear Schrodinger-type equation governing the slow modulation of the wave amplitude is derived. The effects of dust temperature, dust charge variation, ion deviation from Maxwellian equilibrium (nonthermality) and constituent species' concentration on the modulational instability of DA waves are examined. It is found that these parameters modify significantly the oblique modulational instability domain in the k-theta plane. Explicit expressions for the instability rate and threshold have been obtained in terms of the dispersion laws of the system. The possibility and conditions for the existence of different types of localized excitations are also discussed. The findings of this investigation may be useful in understanding the stable electrostatic wave packet acceleration mechanisms close to the Moon, and also enhances our knowledge on the occurrence of instability associated to pickup ions around unmagnetized bodies, such as comets, Mars, and Venus.

Improving the Fuel Economy of a Tuned 600cc FSAE Engine

To maintain its relevance, motorsport cannot be exempt from
the trend of increasing fuel economy. This bears obvious
competitive benefits as well, either in decreasing the
frequency of pit stops or the mass of fuel carried. Given the
increased points weighting of fuel economy for the Formula
Student (FS) competition, a complete analysis was performed
on the Queen's Formula Racing 600cc motorcycle engine in
preparation for the 2011 competition.
The criteria for such high performance fuel economy differ to
a degree from most mass transportation counterparts and were
divided into three distinct regimes; full load, part load and no
load conditions.
Full load positions naturally demand maximum torque for
performance but that does not imply that fuel savings cannot
be made whilst preserving this. The point at which maximum
torque is produced with minimum air -fuel ratio, Leanest
mixture for Best Torque (LBT), was therefore sought and
mapped for full load.
At part load, torque is less of a concern, and maintaining a
sustainable engine temperature and transient response become
more important. With decreasing AFR, engine temperatures
can rise dramatically so temperatures were measured close to
the exhaust port for a wide range of air-fuel ratios.
Competition track data was analysed to highlight key part load
operating regions and these were mapped according to
measured safe temperature limits. Torque response to a step
throttle change was also measured to ensure suitable engine
transient performance was maintained.
At no load conditions, with low engine speed only idle
conditions need to be satisfied. In the situation where the
engine is still at high speed without load, the engine is being
motored and no fuel is required. An overrun fuel cut was
employed to reflect this giving significant fuel savings. The
effect on torque and engine pickup was measured.
Modifications were also made to the fuel injector location to
improve fuel mixing and evaporation at this lower air flow
condition.
These mapping regimes were implemented and tested using
fully transient lap simulations using competition track data
and a four quadrant AC engine dynamometer. The experiment
indicated a reduction in fuel consumption for 22 laps of the FS
track from 5.08litres to 3.67litres, around 27% in total. The
actual fuel used at the 2011 competition was 3.6 litres while
placing 8th in the endurance event, further validating the
benefits of these mapping regimes.

Fasciola hepatica:the effect of the microfilament inhibitor cytochalasin B on the ultrastructure of the adult fluke

The effect of the microfilament inhibitor cytochalasin B (10 and 100 micrograms/ml) on the ultrastructure of adult Fasciola hepatica was determined in vitro by scanning and transmission electron microscopy (SEM, TEM) using both intact flukes and tissue-slice material. SEM revealed that initial swelling of the tegument led to surface blebbing and limited areas of sloughing after 24 h treatment at 100 micrograms/ml. In the tegumental syncytium, basal accumulations of secretory bodies (especially T2s) were evident in the earlier time periods but declined with longer incubations, until few secretory bodies remained in the syncytium overall. Blebbing of the apical plasma membrane and occasional areas of breakdown and sloughing of the tegument were observed over longer periods of treatment at 100 micrograms/ml. In the tegumental cell bodies, the Golgi complexes gradually decreased in size and activity, and few secretory bodies were produced. In the later time periods, the cells assumed abnormal shapes, the cytoplasm shrinking in towards the nucleus. In the vitelline follicles, a random dispersion of shell protein globules was evident within the intermediate-type cells, rather than their being organized into distinct shell globule clusters. Disruption of this process was more severe at the higher concentration of 100 micrograms/ml and again was more evident in tissue-slice material. In the latter, after prolonged (12 h) exposure to cytochalasin B, the intermediate and mature vitelline cells were filled with loosely packed and expanded shell globule clusters, containing few shell protein globules. The mature vitelline cells continued to lay down "yolk" globules and glycogen deposits. Disruption of the network of processes from the nurse cells was evident at the higher concentration of cytochalasin. Spaces began to appear between the vitelline cells and grew larger with progressively longer incubation periods, and the cells themselves assumed abnormal shapes. A number of binucleate stem cells were observed in tissue-slice material at the longest incubation period (12 h).

Food availability affects the maternal transfer of androgens and antibodies into eggs of a colonial seabird.

Mothers can improve the quality of their offspring by increasing the level of certain components in their eggs. To examine whether or not mothers increase deposition of such components in eggs as a function of food availability, we food-supplemented black-legged kittiwake females (Rissa tridactyla) before and during egg laying and compared deposition of androgens and antibodies into eggs of first and experimentally induced replacement clutches. Food-supplemented females transferred lower amounts of androgens and antibodies into eggs of induced replacement clutches than did non-food-supplemented mothers, whereas first clutches presented no differences between treatments. Our results suggest that when females are in lower condition, they transfer more androgens and antibodies into eggs to facilitate chick development despite potential long-term costs for juveniles. Females in prime condition may avoid these potential long-term costs because they can provide their chicks with more and higher quality resources.

Hunter's panoramic guide from Niagara Falls to Quebec /

Reissue of original ed.: Boston : J.P. Jewett ; Cleveland : H.P.B. Jewett, 1857. Cf. TPL 3664.

Une heuristique à grand voisinage pour un problème de confection de tournée pour un seul véhicule avec cueillettes et livraisons et contrainte de chargement

Dans ce mémoire, nous présentons un nouveau type de problème de confection de tour- née pour un seul véhicule avec cueillettes et livraisons et contrainte de chargement. Cette variante est motivée par des problèmes similaires rapportés dans la littérature. Le véhi- cule en question contient plusieurs piles où des colis de hauteurs différentes sont empilés durant leur transport. La hauteur totale des items contenus dans chacune des piles ne peut dépasser une certaine hauteur maximale. Aucun déplacement n’est permis lors de la li- vraison d’un colis, ce qui signifie que le colis doit être sur le dessus d’une pile au moment d’être livré. De plus, tout colis i ramassé avant un colis j et contenu dans la même pile doit être livré après j. Une heuristique à grand voisinage, basé sur des travaux récents dans le domaine, est proposée comme méthode de résolution. Des résultats numériques sont rapportés pour plusieurs instances classiques ainsi que pour de nouvelles instances.

Problèmes de tournées de véhicules avec contraintes de chargement

Cette thèse s’intéresse aux problèmes de tournées de véhicules où l’on retrouve des contraintes de chargement ayant un impact sur les séquences de livraisons permises. Plus particulièrement, les items placés dans l’espace de chargement d’un véhicule doivent être directement accessibles lors de leur livraison sans qu’il soit nécessaire de déplacer d’autres items. Ces problèmes sont rencontrés dans plusieurs entreprises de transport qui livrent de gros objets (meubles, électroménagers). Le premier article de cette thèse porte sur une méthode exacte pour un problème de confection d’une seule tournée où un véhicule, dont l’aire de chargement est divisée en un certain nombre de piles, doit effectuer des cueillettes et des livraisons respectant une contrainte de type dernier entré, premier sorti. Lors d’une collecte, les items recueillis doivent nécessairement être déposés sur le dessus de l’une des piles. Par ailleurs, lors d’une livraison, les items doivent nécessairement se trouver sur le dessus de l’une des piles. Une méthode de séparation et évaluation avec plans sécants est proposée pour résoudre ce problème. Le second article présente une méthode de résolution exacte, également de type séparation et évaluation avec plans sécants, pour un problème de tournées de véhicules avec chargement d’items rectangulaires en deux dimensions. L’aire de chargement des véhicules correspond aussi à un espace rectangulaire avec une orientation, puisque les items doivent être chargés et déchargés par l’un des côtés. Une contrainte impose que les items d’un client soient directement accessibles au moment de leur livraison. Le dernier article aborde une problème de tournées de véhicules avec chargement d’items rectangulaires, mais où les dimensions de certains items ne sont pas connus avec certitude lors de la planification des tournées. Il est toutefois possible d’associer une distribution de probabilités discrète sur les dimensions possibles de ces items. Le problème est résolu de manière exacte avec la méthode L-Shape en nombres entiers.