Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.     

Recombinant Bone Morphogenetic Proteins: Novel Substances for Enhancing Bone Healing

Bone morphogenetic proteins (BMPs) are differentiative factors whose principal function is to induce transformation of undifferentiated mesenchymal cells into chondroblasts and osteoblasts in a dose-dependent manner. Bone morphogenetic proteins have been isolated postnatally in mammals from bone matrix, periosteal cells, mesenchymal cells of marrow stroma, tooth anlagen, and cells of osteosarcoma and chondrosarcoma. Distribution in additional embryonic tissues implies a broader organogenic function. Bone morphogenetic proteins are the only differentiative factors able to singularly induce de novo bone formation in vitro and in vivo. Recombinant DNA technology allows their production in large and highly purified quantities. The BMPs' osteoinductive ability has been shown with a variety of carriers including collagens and polymers at heterotopic and orthotopic sites in a wide range of species. They are presently being readied for clinical use as alternatives to bone grafts. Other potential applications include use as pulp capping agents, promoters of implant osteointegration and soft tissue reunion with bone, treatments for nonadaptive bone disease, and implants for use with mitotically expanded skeletal stem cell populations. Errors in the genetic coding of BMPs may manifest as clinical disease entities.     

Clinical cytogenetics in equine stud practice: Consideration of the present situation and future application

The situation in Thoroughbred breeding in relation to cytogenetics is considered and it is indicated where screening of horses will be useful in practice. The techniques involved are explained and karyotypes are illustrated. The present state of knowledge in horse cytogenetics is reviewed. The types of abnormality are described in relation to phenotype. Future promise is discussed in relation to the human field, concluding that with the refined methods now available a wider rangeof meaningful aberrations will be uncovered.     

Palmar Digital Neurectomy in Horses 57 Cases (1984–1990)

The medical records of 57 horses that had palmar digital neurectomy performed between 1984 and 1990 were reviewed. Neurectomies were performed either by transection and elec-trocoagulation (47 horses) or by the guillotine technique (10 horses). Middle-aged geldings, Quarter Horses, and Thoroughbreds were significantly over-represented when compared with the hospital population. Horses used as hunter/jumpers also appeared to be over-represented. Complications occurred in 17 (34%) of the 50 horses for which follow-up information was obtained. Recurrence of heel pain was the most common complication (14 horses). Palpable painful neuromas were detected in three horses. One year after neurectomy, 74% of the horses were sound; this decreased to 63% after 2 years.     

The Effect of Occlusive and Semi-Occlusive Dressings on the Healing of Acute Full-Thickness Skin Wounds on the Forelimbs of Dogs

This project compared the effects of hydrocolloid (HC) and hydrogel (HG) occlusive dressings and a polyethylene (PE) semi-occlusive dressing on the healing of acute full-thickness skin wounds on the forelimbs of 10 dogs. All treatments resulted in a similar degree of healing at postoperative days 4 and 7. No significant differences existed in the number of wounds that were more than 90% healed at postoperative day 28 between the group treated with the HG dressing and the group treated with the PE dressing. There were significantly fewer wounds more than 90% healed at postoperative day 28 in the group treated with the HC dressing. Wounds under the HG dressing had the largest mean percentage of contraction at postoperative days 21 and 28. Wounds under the HG dressing also had the largest contraction/re-epithelialization ratio (postoperative days 21 and 28) compared with wounds under the PE and HC dressings. Wounds under the PE dressing had a significantly higher mean percentage of re-epithelialization than wounds under both occlusive dressings on postoperative days 14, 21, and 28. Wounds under the two occlusive dressings had exuberant granulation tissue present more often than wounds under the PE dressing. The two occlusive dressings had significantly higher bacterial counts on wounds compared with wounds under the PE dressing; analysis of variance (ANOVA), P = .0008. Wounds under the HC dressing showed the poorest healing in all parameters.     

Evaluation of auditory evoked potentials to predict depth of anaesthesia during fentanyl/fluanisone−midazolam anaesthesia in rats

Objectives To assess a method for monitoring depth of anaesthesia using components of middle latency auditory evoked potential (AEP) waveforms during anaesthesia with fentanyl/fluanisone and midazolam. Study design Prospective observational study. Animals Five female Wistar rats weighing between 210 and 250 g. Methods Implanted electrodes were used to record AEPs in animals receiving five doses of anaesthetic. Recordings were made at 5 minutes post‐injection (deep anaesthesia; no pedal withdrawal response, PWR) and then at 25 minutes (light anaesthesia; strong PWR). Responses showed five characteristic peaks occurring at 11, 14, 23, 42 and 68 ms that were measured for latency of occurrence and peak amplitude. Results Auditory evoked potential peaks P14, N23 and P42 were increased significantly in latency with successive anaesthetic injections [avg. F(1,4) = 12.53, p < 0.001; avg. F(1,4) = 10.6, p < 0.001; avg. F(1,4) = 3.9, p = 0.02, respectively]. Peak N23 showed a significant reduction in latency during the 20 minute recovery period following both the first and second anaesthetic injections (t(3) = 7.52, p = 0.005; t(4) = 5.17, p = 0.007, respectively). Peak P42 occurred significantly earlier 20 minutes following the second anaesthetic injection (t(4) = 4.75, p = 0.009). The mean overall depth of anaesthesia assessed using PWR scores was significantly correlated with the mean latency of peak N23, such that as the strength of PWR increased, N23 occurred significantly earlier (r = ?0.99, p = 0.01). The amplitude difference between peaks N23 and P42 increased after the second and third drug administrations [avg. F(1,4) = 10.65, p = 0.031 and avg. F(1,4) = 11.24, p = 0.028, respectively]. Conclusion The characteristics of these peaks, and in particular latency of peak N23, may provide a useful tool for assessing depth of anaesthesia produced by this, and possibly other anaesthetic agents.