Effect of Dimethyl Sulfoxide on Cell Cycle Synchronization of Goldfish Caudal Fin Derived Fibroblasts Cells

Several studies have previously been conducted regarding cell cycle synchronization in mammalian somatic cells. However, limited work has been performed on the control of cell cycle stages in the somatic cells of fish. The aim of this study was to determine the cell cycle arresting effects of several dimethyl sulfoxide (DMSO) concentrations for different times on different cell cycle stages of goldfish caudal fin‐derived fibroblasts. Results demonstrated that the cycling cells or control group (68.29%) yields significantly higher (p < 0.05) arrest in G0/G1 phase compared with the group treated for 24 h with different concentrations (0.5%, 1.0% or 1.5%) of DMSO (64.88%, 65.70%, 64.22% respectively). The cell cycle synchronization in the treatment of cells with 1.0% DMSO at 48 h (81.14%) was significantly higher than that in the groups treated for 24 h (76.82%) and the control group (77.90%). Observations showed that treatment of DMSO resulted in an increase in the proportion of cells at G0/G1 phase for 48 h of culture. However, high levels of apoptotic cells can be detected after 48 h of culture treated with 1% concentration of DMSO.     

Kinetics of Plasma Cell‐Free DNA and Creatine Kinase in a Canine Model of Tissue Injury

Background

Cell‐free DNA (cfDNA) comprises short, double‐stranded circulating DNA sequences released from damaged cells. In people, cfDNA concentrations correlate well with disease severity and tissue damage. No reports are available regarding cfDNA kinetics in dogs.

Objectives/Hypothesis

Cell‐free DNA will have a short biological half‐life and would be able to stratify mild, moderate, and severe tissue injury. Our study aims were to determine the kinetics and biological half‐life of cfDNA and to contrast them with those of creatine kinase (CK).

Animals

Three groups of 10 dogs undergoing open ovariohysterectomy, surgery for cranial cruciate ligament rupture (CCLR), or hemilaminectomy.

Methods

Plasma for cfDNA and CK analysis was collected at admission, at induction of anesthesia, postsurgery (time 0) and at 6, 12, 24, 36, 48, 60, and 72 hours after surgery.

Results

The biological half‐life of plasma cfDNA and CK were 5.64 hours (95% confidence interval [CI 95], 4.36–7.98 hours) and 28.7 hours (CI95, 25.3–33.3 hours), respectively. In the hemilaminectomy group, cfDNA concentrations differed significantly from admission at 6–12 hours after surgery. Creatine kinase activity differed among the surgical groups and reached a peak 6 hours after surgery. In the ovariohysterectomy and CCLR groups, plasma CK activity 72 hours after surgery did not differ from admission activity of the ovariohysterectomy group. In contrast, in the hemilaminectomy group, plasma CK activity after 72 hours did not return to the ovariohysterectomy group admission activity.

Conclusions and Clinical Importance

Plasma CK activity has a longer biological half‐life than previously thought. In contrast to plasma CK activity, cfDNA has a short half‐life and could be a useful marker for peracute severe tissue injury.     

Histological response of resistant and susceptible white spruce to simulated white pine weevil damage

The traumatic wound response of families of white spruce, Picea glauca (Moench) Voss, resistant or susceptible to the white pine weevil, Pissodes strobi (Peck), were compared after simulated weevil damage. Leaders from 331 trees were wounded just below the apical bud in the spring, coinciding with the natural time of weevil oviposition. A portable 1-mm diameter drill was used to drill 24 holes per leader. Leaders were removed in the fall and examined for evidence of traumatic resin canal formation. Drilled trees had a traumatic wound response 8 times greater than that of undrilled trees; however, undrilled trees also formed some resin canals in response to unknown causes. In the drilled trees, the traumatic wound response extended into the lower part of the leader, where it could possibly affect older larvae. Trees from resistant families responded with greater intensity than trees from susceptible families, by producing multiple rings of traumatic resin canals. Trees from resistant families also responded more rapidly than trees from susceptible families based on number of cells to the first ring of traumatic resin canals. Trees from some resistant families exhibited no traumatic resin canal formation, showing considerable within-family variation and suggesting that other resistance mechanisms might be important. In the year after drilling, there was a reduction in tree diameter growth and trees suffered a reduction in constitutive resin canals in the bark, which suggests some energetic cost of traumatic resin production. There was no indication that the extent of constitutive defenses, as measured by density of cortical resin canals before wounding, was related to the ability to produce traumatic resin canals. Screening trees based on their capacity to produce traumatic resin canals may be useful in selecting genotypes resistant to white pine weevil.     

The evaluation of endophyte toxin residues in sheep fat

Abstract

AIM: To monitor changes in concentrations of lolitrem B and epoxy-janthitrems in the fat of sheep grazing perennial ryegrass infected with wild-type- and AR37-endophyte, respectively, during the time of year when ryegrass staggers would be expected to be observed.

METHODS: Ten 5-month-old lambs with no previous exposure to endophytes were grazed on either wild-type (containing lolitrem B, n = 5) or AR37 (containing epoxy-janthitrems, n = 5) endophyte-infected perennial ryegrass (Lolium perenne L.) pastures between October 2008 and June 2009. Animals were regularly assessed for ryegrass staggers using the Keogh scale (0 = no signs, 5 = severe tremors). When a score of > 3.5 was observed animals were removed from the treatment pastures for 1 month. Fat biopsy samples were taken from each animal at approximately monthly intervals and analysed for endophyte metabolites using high performance liquid chromatography (HPLC) methods developed during this study. Regular herbage samples were also taken and concentrations of endophyte metabolites measured.

RESULTS: Efficient and reproducible methods to analyse both lolitrem B and epoxy-janthitrems in fat were developed. Concentrations of lolitrem B and epoxy-janthitrems in herbage and in sheep fat increased from late November to peak in mid-February. Ryegrass staggers was observed in both groups of sheep at this time. Following 1 month of grazing non-infected pasture mean concentrations in fat of lolitrem B decreased by 43% from 61.8 to 35.3 ppb, and of epoxy-janthitrems by 38% from 1032.0 to 639.5 ppb. Maximum concentrations in herbage of epoxy-janthitrems (35.7 ppm) were higher than of lolitrem B (3.4 ppm), but signs of staggers were less severe in sheep grazing pasture containing the former compared with the latter (median Keogh scores in late February were 2 and 3, respectively), consistent with epoxy-janthitrems being low potency toxins.

CONCLUSION: This study demonstrated that concentrations of epoxy-janthitrems and lolitrem B in sheep fat increased quickly during the initial phase of the study when concentrations in pasture increased, and decreased when animals were removed from pastures containing these compounds. These data will be used in the risk assessment of the endophyte metabolites.     

Stratospheric Mean Ages and Transport Rates from Observations of Carbon Dioxide and Nitrous Oxide

Measurements of stratospheric carbon dioxide (CO2) and nitrous oxide (N2O) concentrations were analyzed to investigate stratospheric transport rates. Temporal variations in tropospheric CO2 were observed to propagate into the stratosphere, showing that tropospheric air enters the lower tropical stratosphere continuously, ascends, and is transported rapidly (in less than 1 month) to both hemispheres. The mean age A of stratospheric air determined from CO2 data is approximately 5 years in the mid-stratosphere. The mean age is mathematically equivalent to a conserved tracer analogous to exhaust from stratospheric aircraft. Comparison of values for A from models and observations indicates that current model simulations likely underestimate pollutant concentrations from proposed stratospheric aircraft by 25 to 100 percent.     

Measurement of Parity Nonconservation and an Anapole Moment in Cesium

The amplitude of the parity-nonconserving transition between the 6S and 7S states of cesium was precisely measured with the use of a spin-polarized atomic beam. This measurement gives Im(E1pnc)/beta = -1.5935(56) millivolts per centimeter and provides an improved test of the standard model at low energy, including a value for the S parameter of -1.3(3)exp (11)theory. The nuclear spin-dependent contribution was 0.077(11) millivolts per centimeter; this contribution is a manifestation of parity violation in atomic nuclei and is a measurement of the long-sought anapole moment.     

The possible adaptive advantages of terrestrial egg deposition in some fluvial diadromous galaxiid fishes (Teleostei: Galaxiidae)

Several diadromous New Zealand and Australian species of Galaxias are now known, or suspected, to deposit their eggs amongst riparian vegetation or substrates either supratidally in estuaries or in forested streams in locations that are only temporarily submerged by elevated water levels. The eggs develop in a humid atmosphere and hatch when the egg deposition sites are resubmerged; a significant role for agitation in stimulating hatching seeming likely. There are risks from the eggs becoming dehydrated, and also from a failure by water to resubmerge the eggs before they have exhausted their energy resources. Hatching is triggered by elevated flows, perhaps being an outcome of agitation of the eggs. Elevated flows may also increase the rate of downstream transport of the larvae, facilitating survival during dispersal to sea from spawning sites in streams that may be long distances inland. Hatching during flood events may favour survival of the larvae because turbid flows may provide ‘cover’ for the larvae as they emigrate to sea. Risks from egg predation by aquatic predators may be replaced by risks from terrestrial predators.