Postreplicative formation of cohesion is required for repair and induced by a single DNA break

Sister-chromatid cohesion, established during replication by the protein complex cohesin, is essential for both chromosome segregation and double-strand break (DSB) repair. Normally, cohesion formation is strictly limited to the S phase of the cell cycle, but DSBs can trigger cohesion also after DNA replication has been completed. The function of this damage-induced cohesion remains unknown. In this investigation, we show that damage-induced cohesion is essential for repair in postreplicative cells in yeast. Furthermore, it is established genome-wide after induction of a single DSB, and it is controlled by the DNA damage response and cohesin-regulating factors. We thus define a cohesion establishment pathway that is independent of DNA duplication and acts together with cohesion formed during replication in sister chromatid-based DSB repair.     

Retrospective comparison of three locoregional techniques for pelvic limb surgery in dogs

ObjectiveTo retrospectively compare the efficacy and duration of effect of three commonly used locoregional blocks in dogs undergoing pelvic limb orthopaedic surgery.Study designRetrospective clinical study.AnimalsA total of 236 dogs that underwent pelvic limb surgery and were administered a locoregional technique.MethodsA total of 236 hospital records were reviewed and 230 included in statistical analysis. Dogs were grouped as following: electrostimulation-guided pre-iliac femoral and sciatic nerve block (group PFS, n = 70); ultrasound-guided saphenous and sciatic nerve block (group SS, n = 76); or lumbosacral epidural (group EPI, n = 84). In group EPI, bupivacaine 0.5% or ropivacaine 0.75% was used with morphine. Dogs were pain scored (short form of the Glasgow Composite Measure Pain Scale) hourly following recovery from anaesthesia. Analysed data included: time to first postoperative dose of methadone, pain score at that time, intraoperative rescue analgesia, intraoperative hypotension and ability to walk and urinate overnight. Separate analyses were performed including all pelvic limb surgeries and including only elective stifle surgeries. Kruskal–Wallis and Mann–Whitney tests were performed. A p value < 0.05 was considered significant. The median (range) is reported.ResultsFor all pelvic limb surgeries, the time to first postoperative methadone was 530 (110–1337), 440 (140–1030) and 466 (135–1094) minutes in groups EPI, PFS and SS, respectively, and was not significantly different. Postoperatively, 10/84, 15/70 and 12/76 dogs in groups EPI, PFS and SS, respectively, did not require methadone (nonsignificant). Significantly fewer dogs in group EPI (18%) required intraoperative rescue analgesia compared with group SS (38%), but not compared with PFS (30%). Significantly more dogs in group EPI had hypotension intraoperatively (30%) and urinary retention postoperatively (62%).Conclusions and clinical relevanceIntraoperative analgesia may be superior with EPI than SS for some surgeries of the pelvic limb, but not for stifle surgeries. All three techniques provided similar requirement for postoperative analgesia, but EPI caused higher incidence of intraoperative hypotension and postopertive urinary retention.     

Molecular confirmation of candidate Hsp70 gene associated with heat tolerance in BC3F2 advanced backcross lines and their phenotypic resemblance with recurrent chilli Kulai

ABSTRACT

Domestic production of chilli in Malaysia is insufficient which accounted for 52% self-sufficiency level. Poor performance of local varieties under high temperatures above 42°C is one of the major factors affecting chilli production. This study was conducted with the objective of introgress heat-tolerant gene (Hsps) from AVPP0702 donor into Kulai using marker-assisted backcrossing. A total of 68 SSR markers, including Hsp linked markers that showed polymorphism between the parents were used to assess the generation of backcrossing; BC1F1, BC2F1, BC3F1 and BC3F2 and the average RPG percentage of the recurrent parent was found to be 81, 90, 95 and 97%, respectively. The pattern of Hsp expression in the backcross generations was similar to the donor parent (upregulated with more than 4-fold increase). Twelve improved heat tolerant chilli lines were developed. Most of the morphological and agronomical traits were recovered in the selected improved heat-tolerant genotypes from Kulai such as plant height, number of days to 50% flowering, number of fruits, fruit length and total fruit yield per plant. Improved high-yielding heat-tolerant chilli lines showed tolerance to high temperature as well as did not express any negative effect on agronomic traits in comparison with Kulai variety.     

Endogenous fat losses and true and apparent fat digestibility in adult and growing dogs fed diets containing poultry offal fat

The objective of this study was to evaluate the coefficient of total tract apparent digestibility (CTTAD) of dietary nutrients, endogenous fat loss (EFL), dietary metabolizable energy (ME) content, coefficient of true digestibility (CTD) of fat and faecal parameters of dogs. A total of 16 puppies and 8 adult dogs were distributed according to a double Latin square experimental design. A basal diet was formulated, to which increasing levels (60, 120 and 180 g/kg) of poultry offal fat oil were added. EFL was estimated using hyperbolic and linear regression as a function of LW/day and dry matter (DM) intake, respectively, and the CTD of fat was calculated. The CTTAD of acid-hydrolysis ether extract (AHEE) and ME content increased as dietary poultry offal fat level increased both in adult dogs and in puppies (p < .05). The CTTAD of DM and crude protein were higher in adult dogs than in puppies. EFL was estimated by hyperbolic regression as y = 98.1094 − 13.6202/x and y = 99.1280 − 26.1366/x, where y = CTTAD of AHEE (%) and x = AHEE intake/kg BW, for adult dogs and puppies, respectively, which yielded EFL values of 138 and 262 mg AHEE per kg BW per day for adult dogs and puppies respectively. The EFL estimated by linear regression was 7.89 g and 8.97 g/kg DM intake for adult dogs and puppies respectively. The CTD estimated by both methodologies was higher in puppies than in adults (p < .05). Lower faecal DM and score, and higher ammonia production were obtained in puppies than adult dogs (p < .05). The results indicate good fat utilization and that fat digestibility is underestimated, as the CTTAD of AHEE does not take into account EFL, particularly at low-fat inclusion levels and in growing dogs, which have greater EFL than adult dogs.     

Toxicokinetics of mercury in blood compartments and hair of fish-fed sled dogs

Background

Understanding mercury (Hg) distribution in blood and the importance of hair as an excretory pathway is critical for evaluating risk from long term dietary Hg exposure. The major objective of this study was to characterize changes in total Hg concentrations in specific blood compartments and hair over time due to long term piscivory.

Methods

Eight sled dogs (Canis lupus familiaris) were fed either a fish and kibble diet (n = 4), or a fish-free control diet (n = 4) for 12 weeks. Concentrations of Hg were monitored throughout the exposure period, and for 10 weeks post exposure, until Hg concentrations in all blood compartments of one of the exposed dogs dropped below detection limit. Additionally, foreleg hair was sampled during acclimation and weeks 0 and 12.

Results

Hg was detected primarily in whole blood and packed cells, although it was sporadically detected at low concentrations in plasma and serum in two of the fish fed dogs. Dogs ingested an estimated average of 13.4 ± 0.58 μg Hg per kg body weight per day. Hg was detectable in whole blood and packed cells within a week of exposure. Detected concentrations continued to rise until plateauing at approximately 3-6 weeks of exposure at a mean of 9.2 ± 1.97 ng/g (ppb) in whole blood. Hg concentration decreased post exposure following 1st order elimination. The mean half-life (t_1/2) in whole blood for Hg was 7 weeks. Mean Hg in hair for the fish-fed dogs at week 12 was 540 ± 111 ppb and was significantly greater (about 7-fold) than the Hg hair concentration for the control dogs. The hair to blood ratio for Hg in fish-fed dogs was 59.0 ± 7.6:1.

Conclusions

This study found the sled dog model to be an effective method for investigating and characterizing blood Hg distribution (whole blood, serum, plasma, packed cells) and toxicokinetics associated with a piscivorous diet, especially for Hg-exposed fur bearing mammals (such as polar bears). Although hair excretion and hair to blood Hg ratios were not similar to human concentrations and ratios, the sled dog toxicokinetics of Hg in blood, was more similar to that of humans than traditional laboratory animals (such as the rat).     

Effect of position and orientation of leaflet explants with respect to plant growth regulators on micropropagation of Zamioculcas zamiifolia Engl. (ZZ)

Micropropagation studies on Zamioculcas zamiifolia Engl. (ZZ) as to the position and orientation of leaflet explants and plant growth regulators were carried out. Explants consisted of leaflet lamina from the basal or apical part of the leaflet with or without petiolule or midrib that were placed vertically into the medium except for the explants with midrib from the basal part of the leaflet that were placed horizontally as well. The explants were cultured on solid Murashige and Skoog medium (MS) with 30 g l⁻¹ sucrose, supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at 2 or 4 mg l⁻¹ and 6-benzyladenine (BA) at 0 or 4.44 μM in all (four) possible combinations, or with 1-naphteleneacetic acid (NAA) at 0 or 5.38 μM and BA at 0 or 4.44 μM in all (four) possible combinations (establishment medium). The morphogenic response was direct from all types of leaflet explants and varied only with respect to different plant growth regulators of the medium: 2,4-D combined or not with BA formed somatic embryo-like structures; NAA alone produced tubers and roots; BA alone resulted mainly in leaves; NAA combined with BA produced mainly roots. The intensity of the response varied accordingly to the explant type and orientation. Explants with petiolule or midrib from the basal part of the leaflet showed the highest morphogenic response compared to explants without petiolule or midrib or to explants from the apical part of the leaflet, in all the plant growth regulator combinations used. Explants with midrib from the basal part of the leaflet placed vertically into the media showed higher morphogenic response compared to those placed horizontally on the medium surface. With the objective to regenerate plantlets, explants were subcultured on MS with NAA and BA at various concentrations based on the explant response in the establishment medium, taking into consideration the initial explant type. The initial explant type did not affect the response in the subculture. Most plantlets (a tuber with roots and one leaf with one pair of leaflets) were produced by explants with embryo-like structures induced in a medium with only 2,4-D. Explants with tubers induced in a medium with NAA gave plantlets at 65–85% when subcultured in a medium with 4.44 μM BA alone or in combination with 2.69 μM NAA. Explants with leaves induced in a medium with BA and explants with roots induced in a medium with NAA and BA gave plantlets at low percentages (20–40%). The best response was produced by explants with embryo like structures induced in a medium with only 2,4-D which gave plantlets at 100% when subcultured in the medium with 2.69 μM NAA and 2.22 μM BA. Plantlets raised in different treatments were transplanted ex vitro after 22 weeks and exhibited 80–100% survival.     

Ceylon-Malaria

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