Effects of EGTA and cytochalasin-B during freezing and vitrification of immature and mature bovine and rhesus monkey (M. mulatta) oocytes.

Cryopreservation procedures have been shown to be extremely disruptive to the plasma membrane and intracellular organization of mammalian oocyte. Cytochalasin-B (Cyt-B) or EGT A may stabilize oocyte cytoskeletal elements prior to Cryopreservation and make the plasma membrane less rigid and more elastic to avoid injury during the osmotic stresses of freezing. Experiments were conducted to assess the morphological viability and developmental potential of bovine and rhesus monkey oocytes roter exposure to Cyt-B or EGTA prior to freezing and vitrification. ln the bovine experiments: The methods of oocyte isolation, in vitro maturation (IVM), in vitro fertilization (IVF) , and embryonic development (IVC) were as described elsewhere (Biol Reprod. 55:333-339, 1996). AIl data were analyzed by ANOV A and Chi-square using "StatMost" for Windows. ln an initial experiment, a group of ilnmature (GV) oocytes and oocytes roter IVM (ova) were randomly selected and pretreated for 5 min with EGTA (O.IM) or Cyt-B (Img/ml) without cryoprotectant (CPA), and another group were equilibrated for 15 min with the CPA (1.5 M ethylene glycol mixed with 0.5 M glycerol in PHS) roter pretreatment with EGTA or Cyt-B. AIl oocytes were diluted and/or washed and immediately used for IVM and/or IVF and IVC. No significant differences in cleavage (C) development and blastocyst (B) hatching among treatments in the GV oocytes group. Percentages of early C, morula (M), B, and expanded B (EB) development ranged 72-82%, 53-65%, 40-47% and 25-35% respectively. Ova treated with EGTA without CPA produced significantly (P<.05) higher C (31/41,75%), M (48.8%), B (40%) and EB (24.4%) than with Cyt-B. ln the slow freezing experiment, a total of 105 GV oocytes and 155 ova were exposed to (a) zero (b EGTA, or ~) Cyt-B before equilibration with CPA + 0.5 sucrose in PHS plus 15% Fetal bovine serum (PBS/FBS). Gametes were loaded into sterile cryovials, cooled at 2°Chnin to -7°C, seeded, cooled at 0.3°Chnin to -32°C, then plunged in LN2. They were thawed.in air (10 sec), then wann water (35°C), and diluted out of CPA in 3 steps (1, 0.5, 0.2 M sucrose). Three replicated showed that although many of the frozen-thawed oocytes/ova survived as judged by morphology (range 70-80%), their developmental potential was severely retarded. C ranged between 0% to 14.3%. ln the vitrification experiment, a total of 135 GV oocytes and 90 ova were grouped as above and vitrified in straws containing a mixture of 3.25M glycerol and 4.5M ethylene glycol plus 0.5 sucrose in PBS/FBS. Here, higher (P<.05) morphological survival rates (98%) foIlowed pretreatment with EGT A. However, the developmental potential was very low among aIl groups ranging between 0% to 17.7%. ln the monkey experiments: The methods of oocyte isolation, IVM and IVF was as described ( Theriogenology 43:362, 1995). A total of 72 GV oocytes and 42 ova were subjected to slow rate freezing and vitrification in groups as described for cow oocytes above. Pretreatment of both GV oocytes or ova with EGTA or Cyt-B prior to freezing or vitrification resulted in 77.6% morphological survival which was significantly (P<.OI) higher than for the control (50%). No differen?s in fertilization and development were detected mnong treatment. Per?ntages of C to 4-?Il stage ranged from 50-67%. Results suggest prefreeze treatment with EGTA and/or Cyt-B may be useful in enhancement of oocyte/ovum cryopreservation.Proceedings Annual Conference of the International Embryo Transfer Society, Nice Acropolis, Nice, France, Jan., 1997

Does rumination mediate the relationship between emotion regulation ability and posttraumatic stress disorder?

Background and objectives: Trauma-related rumination has been suggested to be involved in the maintenance of posttraumatic stress disorder (PTSD). This view has empirically been supported by extensive evidence using cross-sectional, prospective, and experimental designs. However, it is unclear why trauma survivors engage in rumination despite its negative consequences. The current study aimed to explore the hypothesis that low emotion regulation ability underlies trauma-related rumination. Methods: Emotion regulation ability and trauma-related rumination were assessed in 93 road traffic accident survivors 2 weeks post-trauma. In addition, symptom levels of PTSD were assessed at 2 weeks as well as 1, 3, and 6 months follow-up. Results: Emotion regulation ability was significantly related to trauma-related rumination as well as levels of PTSD symptoms. In addition, the association between low emotion regulation ability and PTSD was mediated by rumination. Conclusions: The findings support the view that rumination is used as a dysfunctional emotion regulation strategy by trauma survivors

Calculation of The Band Gap Energy and Study of Cross Luminescence in Alkaline-Earth Dihalide Crystals

The band gap energy as well as the possibility of cross luminescence processes in alkaline-earth dihalide crystals have been calculated using the ab initio Perturbed-Ion (PI) model. The gap is calculated in several ways: as a difference between one-electron energy eigenvalues and as a difference between total energies of appropriate electronic states of the crystal, both at the HF level and with inclusion of Coulomb correlation effects. In order to study the possibility of ocurrence of cross luminescence in these materials, the energy difference between the valence band and the upmost core band for some representative crystals has been calculated. Both calculated band gap energies and cross luminescence predictions compare very well with the available experimental results.Comment: LaTeX file containing 8 pages plus 1 postscript figure. Final version accepted for publication in The Journal of the Physical Society of Japan. It contains a more complete list of references, as well as a more detailed comparison with previous theoretical investigations on the subjec

Quaternary Ammonium Silane-Functionalized, Methacrylate Resin Composition With Antimicrobial Activities and Self-Repair Potential

The design of antimicrobial polymers to address healthcare issues and minimize environmental problems is an important endeavor with both fundamental and practical implications. Quaternary ammonium silane-functionalized methacrylate (QAMS) represents an example of antimicrobial macromonomers synthesized by a sol–gel chemical route; these compounds possess flexible Si–O–Si bonds. In present work, a partially hydrolyzed QAMS co-polymerized with 2,2-[4(2-hydroxy 3-methacryloxypropoxy)-phenyl]propane is introduced. This methacrylate resin was shown to possess desirable mechanical properties with both a high degree of conversion and minimal polymerization shrinkage. The kill-on-contact microbiocidal activities of this resin were demonstrated using single-species biofilms of Streptococcus mutans (ATCC 36558), Actinomyces naeslundii (ATCC 12104) and Candida albicans (ATCC 90028). Improved mechanical properties after hydration provided the proof-of-concept that QAMS-incorporated resin exhibits self-repair potential via water-induced condensation of organic modified silicate (ormosil) phases within the polymerized resin matrix

Mechanical properties of Ti-6Al-4V selectively laser melted parts with body-centred-cubic lattices of varying cell size

Significant weight savings in parts can be made through the use of additive manufacture (AM), a process which enables the construction of more complex geometries, such as functionally graded lattices, than can be achieved conventionally. The existing framework describing the mechanical properties of lattices places strong emphasis on one property, the relative density of the repeating cells, but there are other properties to consider if lattices are to be used effectively. In this work, we explore the effects of cell size and number of cells, attempting to construct more complete models for the mechanical performance of lattices. This was achieved by examining the modulus and ultimate tensile strength of latticed tensile specimens with a range of unit cell sizes and fixed relative density. Understanding how these mechanical properties depend upon the lattice design variables is crucial for the development of design tools, such as finite element methods, that deliver the best performance from AM latticed parts. We observed significant reductions in modulus and strength with increasing cell size, and these reductions cannot be explained by increasing strut porosity as has previously been suggested. We obtained power law relationships for the mechanical properties of the latticed specimens as a function of cell size, which are similar in form to the existing laws for the relative density dependence. These can be used to predict the properties of latticed column structures comprised of body-centred-cubic (BCC) cells, and may also be adapted for other part geometries. In addition, we propose a novel way to analyse the tensile modulus data, which considers a relative lattice cell size rather than an absolute size. This may lead to more general models for the mechanical properties of lattice structures, applicable to parts of varying size

In vitro analysis of the cytotoxicity and the antimicrobial effect of four endodontic sealers

<p>Abstract</p> <p>Introduction</p> <p>The aim of this study was to investigate <it>in vitro </it>the cytotoxicity and antibacterial properties of four different endodontic sealers using human periodontal ligament fibroblast cell proliferation and visual analysis of growth inhibition.</p> <p>Methods</p> <p>A silicone (GuttaFlow), silicate (EndoSequence BC), zinc oxide eugenol (Pulp Canal Sealer EWT) and epoxy resin (AH Plus Jet) based sealer were incubated with PDL fibroblasts (10⁴cells/ml, n = 6) up to 96 h. Cell proliferation (RFU) was determined by means of the Alamar Blue assay. Cell growth and morphology was visualized by means of fluorescent dyes. Possible antibacterial properties of the different sealers were visualized by means of SEM (<it>Enterococcus faecalis; Parvimonas micra</it>).</p> <p>Results</p> <p>Fibroblast proliferation depended on sealer and cultivation time. After 72 and 96 h GuttaFlow and EndoSequence BC showed relatively non-cytotoxic reactions, while Pulp Canal Sealer EWT and AH Plus Jet caused a significant decrease of cell proliferation (p < 0.001). Visualization of cell growth and morphology with various fluorescent dyes supplemented the results. No antibacterial effect of EndoSequence BC to <it>P. micra </it>was found, whereas GuttaFlow showed a weak, Pulp Canal Sealer EWT and AH Plus Jet extensive growth inhibition. Also, no antibacterial effect of GuttaFlow, EndoSequence BC or AH Plus Jet to <it>E. faecalis </it>could be detected.</p> <p>Conclusions</p> <p>These <it>in vitro </it>findings reveal that GuttaFlow and EndoSequence BC can be considered as biocompatible sealing materials. However, prior to their clinical employment, studies regarding their sealing properties also need to be considered.</p

Spinal Cord Injury Causes Sustained Disruption of the Blood-Testis Barrier in the Rat

There is a high incidence of infertility in males following traumatic spinal cord injury (SCI). Quality of semen is frequently poor in these patients, but the pathophysiological mechanism(s) causing this are not known. Blood-testis barrier (BTB) integrity following SCI has not previously been examined. The objective of this study was to characterize the effects of spinal contusion injury on the BTB in the rat. 63 adult, male Sprague Dawley rats received SCI (n = 28), laminectomy only (n = 7) or served as uninjured, age-matched controls (n = 28). Using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI), BTB permeability to the vascular contrast agent gadopentate dimeglumine (Gd) was assessed at either 72 hours-, or 10 months post-SCI. DCE-MRI data revealed that BTB permeability to Gd was greater than controls at both 72 h and 10 mo post-SCI. Histological evaluation of testis tissue showed increased BTB permeability to immunoglobulin G at both 72 hours- and 10 months post-SCI, compared to age-matched sham-operated and uninjured controls. Tight junctional integrity within the seminiferous epithelium was assessed; at 72 hours post-SCI, decreased expression of the tight junction protein occludin was observed. Presence of inflammation in the testes was also examined. High expression of the proinflammatory cytokine interleukin-1 beta was detected in testis tissue. CD68+ immune cell infiltrate and mast cells were also detected within the seminiferous epithelium of both acute and chronic SCI groups but not in controls. In addition, extensive germ cell apoptosis was observed at 72 h post-SCI. Based on these results, we conclude that SCI is followed by compromised BTB integrity by as early as 72 hours post-injury in rats and is accompanied by a substantial immune response within the testis. Furthermore, our results indicate that the BTB remains compromised and testis immune cell infiltration persists for months after the initial injury

Ozone, aerosol, potential vorticity, and trace gas trends observed at high‐latitudes over North America from February to May 2000

Ozone (O3) and aerosol scattering ratio profiles were obtained from airborne lidar measurements on thirty‐eight flights over seven deployments covering the latitudes of 40°–85°N between 4 February and 23 May 2000 as part of the Tropospheric Ozone Production about the Spring Equinox (TOPSE) field experiment. Each deployment started from Broomfield, Colorado, with bases in Churchill, Canada, and on most deployments, Thule Air Base, Greenland. Nadir and zenith lidar O3 measurements were combined with in situ O3 measurements to produce vertically continuous O3 profiles from near the surface to above the tropopause. Potential vorticity (PV) distributions along the flight track were obtained from several different meteorological analyses. Ozone, aerosol, and PV distributions were used together to identify the presence of pollution plumes and stratospheric intrusions. Ozone was found to increase in the middle free troposphere (4–6 km) at high latitudes (60°–85°N) by an average of 4.6 ppbv/mo (parts per billion by volume per month) from about 54 ppbv in early February to over 72 ppbv in mid‐May. The average aerosol scattering ratios at 1064 nm in the same region increased rapidly at an average rate of 0.36/mo from about 0.38 to over 1.7. Ozone and aerosol scattering were highly correlated over the entire field experiment, and PV and beryllium (7Be) showed no significant positive trend over the same period. The primary cause of the observed O3 increase in the mid troposphere at high latitudes was determined to be the photochemical production of O3 in pollution plumes with less than 20% of the increase from stratospherically‐derived O3

LSST: from Science Drivers to Reference Design and Anticipated Data Products

(Abridged) We describe here the most ambitious survey currently planned in the optical, the Large Synoptic Survey Telescope (LSST). A vast array of science will be enabled by a single wide-deep-fast sky survey, and LSST will have unique survey capability in the faint time domain. The LSST design is driven by four main science themes: probing dark energy and dark matter, taking an inventory of the Solar System, exploring the transient optical sky, and mapping the Milky Way. LSST will be a wide-field ground-based system sited at Cerro Pach\'{o}n in northern Chile. The telescope will have an 8.4 m (6.5 m effective) primary mirror, a 9.6 deg$^2$ field of view, and a 3.2 Gigapixel camera. The standard observing sequence will consist of pairs of 15-second exposures in a given field, with two such visits in each pointing in a given night. With these repeats, the LSST system is capable of imaging about 10,000 square degrees of sky in a single filter in three nights. The typical 5$\sigma$ point-source depth in a single visit in $r$ will be $\sim 24.5$ (AB). The project is in the construction phase and will begin regular survey operations by 2022. The survey area will be contained within 30,000 deg$^2$ with $\delta<+34.5^\circ$, and will be imaged multiple times in six bands, $ugrizy$, covering the wavelength range 320--1050 nm. About 90\% of the observing time will be devoted to a deep-wide-fast survey mode which will uniformly observe a 18,000 deg$^2$ region about 800 times (summed over all six bands) during the anticipated 10 years of operations, and yield a coadded map to $r\sim27.5$. The remaining 10\% of the observing time will be allocated to projects such as a Very Deep and Fast time domain survey. The goal is to make LSST data products, including a relational database of about 32 trillion observations of 40 billion objects, available to the public and scientists around the world.Comment: 57 pages, 32 color figures, version with high-resolution figures available from https://www.lsst.org/overvie

Validating the German Version of the Quality of Relationship Inventory: Confirming the Three-Factor Structure and Report of Psychometric Properties

Research on psychosocial influences such as relationship characteristics has received increased attention in the clinical as well as social-psychological field. Several studies demonstrated that the quality of relationships, in particular with respect to the perceived support within intimate relationships, profoundly affects individuals' mental and physical health. There is, however, a limited choice of valid and internationally known assessments of relationship quality in Germany. We report the validation of the German version of the Quality of Relationships Inventory (QRI). First, we evaluated its factor structure in a representative German sample of 1.494 participants by means of confirmatory factor analysis. Our findings support the previously proposed three-factor structure. Second, importance and satisfaction with different relationship domains (family/children and relationship/sexuality) were linked with the QRI scales, demonstrating high construct validity. Finally, we report sex and age differences regarding the perceived relationship support, conflict and depth in our German sample. In conclusion, the QRI is a reliable and valid measurement to assess social support in romantic relationships in the German population