Arthroscopic reattachment of osteochondritis dissecans cartilage flaps of the femoropatellar joint: long-term results

Reasons for performing study: Long‐term efficacy of arthroscopic cartilage reattachment for the treatment of osteochondritis dissecans (OCD) lesions in the equine femoropatellar joint is unknown. Objective: To evaluate radiographic outcome and long‐term performance of horses undergoing OCD reattachment. Hypothesis: Separated OCD cartilage flaps may be reincorporated into the joint surface by reattachment rather than flap removal. Methods: Polydioxanone pins were utilised arthroscopically to reattach OCD lesions in 40 of 44 joints from 27 horses. Cartilage was reattached when it had persisting perimeter continuity, the surface was not deeply fissured or irregular, and the cartilage was not protuberant or extensively mineralised. Bone marrow aspirate concentrate was grafted to additional areas denuded of cartilage or alongside reattached cartilage. Results: Breeds included Thoroughbred (n = 18), Quarter Horse (n = 4), Warmblood (n = 3), Standardbred (n = 1) and Arabian (n = 1). Mean age was 9.7 months. Radiographic lesion length was 1.5–6.3 cm. Reattachment alone was used in 32 of 44 affected joints, a combination of debridement and reattachment in 8 joints and debridement alone in 4 joints. One horse was destroyed due to tendon laceration. Of the remaining 26 horses, mean duration of follow‐up was 15.6 months (range 2 months–12 years). Radiographic resolution of OCD lesions treated with reattachment was significantly improved at 6 months. Twenty horses had long‐term performance data, of which 19 were sound and had reached intended athletic potential. One horse remained lame, and an additional 6 were sound but remained unbroken or were convalescing. Thus, an overall success rate based upon continued soundness in performing horses was 95% (19/20). Conclusion: Cartilage flap reattachment can salvage OCD cartilage by integration with the underlying bone. Potential relevance: Extensive OCD cartilage flaps may be salvaged by reattachment which can result in normal radiographic subchondral bone contour and long‐term athletic performance.     

Evaluation of the effects of long-term storage of bovine ear notch samples on the ability of 2 diagnostic assays to identify calves persistently infected with bovine viral diarrhoea virus

Research aimed at optimising diagnostic laboratory procedures is central to the development of effective bovine viral diarrhoea virus (BVDV) control programmes. BVDV is a single-stranded RNA virus that crosses the placenta to infect foetuses, resulting in reproductive losses due to foetal death or persistently infected calves that die early in life. Persistently infected animals are widely accepted to be the primary reservoir of BVDV and the largest source of infection. This poses important challenges to overall animal/herd health and can cause major losses to the cattle industry. Long-term storage of bovine ear notch samples from calves persistently infected with BVDV may adversely affect the ability of diagnostic assays to detect the virus efficiently. In order to test this hypothesis, ear notch samples from 7 animals were divided into 2 groups. One set was subjected to prompt formalin fixation and the other set stored either as fresh samples without preservatives at -2 degrees C, or soaked overnight in phosphate buffered saline followed by freezing of the supernatant fluid at -2 degrees C. Frozen ear notches and ear notch supernatant yielded positive results with an antigen-capture, enzyme linked immunosorbent assay (AC-ELISA) for the duration of the study (6 months) and optical density (OD) values remained significantly within range. There was no significant difference between storing fresh ear notch samples or PBS at -2 degrees C. However, positive immunohistochemistry (IHC) staining on formalin fixed ear notches started to fade between Day 17 and Day 29 when stored at room temperature. It was concluded that fresh ear notches could safely be stored at -2 degrees C for a period of 6 months prior to testing for BVD viral antigens.     

Delayed harvest improves red blush development and quality of ‘Cripps Pink’ apple

The poor red blush development on ‘Cripps Pink’ (Malus x domestica Borkh.) apple fruit at commercial maturity (CM) reduces the economic return to apple growers in warmer regions of the world. The effects of delayed harvest maturity up to 6 weeks following CM on the development of red blush and fruit quality of ‘Cripps Pink’ apple were investigated at two locations in Western Australia during 2003 and 2004. The red blush, export-grade fruit, total anthocyanin, cyanidin 3-galactoside, chlorogenic acid, catechin, and epicatechin of apple fruit skin tissues increased with advancement of harvest maturity at both locations in both years, whereas hue angle, concentration of phloridzin, fruit firmness, and titratable acidity (TA) of apple fruit tissues decreased with delayed harvest. The internal ethylene concentration was elevated with the delayed harvest. The concentrations of quercetin 3-galactoside, quercetin 3-arabinoside, quercetin 3-rutinoside, quercetin 3-rhamnoside, and quercetin 3-glucoside increased up to 2 weeks following CM, and then declined in 2003 at both locations and at Perth Hills in 2004. Delayed harvest resulted in improved development of red blush on the fruit surface, accumulation of total anthocyanin, cyanidin 3-galactoside, chlorogenic acid, catechin, epicatechin, and quercetin glycosides in fruit skin, SSC:TA ratio and reduced fruit firmness possibly due to the increased ethylene production.     

Increased resistance to fusarium wilt in transgenic tobacco lines co‐expressing chitinase and wasabi defensin genes

Marker‐free transgenic tobacco (Nicotiana tabacum) lines containing a chitinase (ChiC) gene isolated from Streptomyces griseus strain HUT 6037 were produced by Agrobacterium‐mediated transformation. One marker‐free transgenic line, TC‐1, was retransformed with the wasabi defensin (WD) gene, isolated from Wasabia japonica. Of the retransformed shoots, 37% co‐expressed the ChiC/WD genes, as confirmed by western and northern analyses. Southern blot analysis showed that no chromosomal rearrangement was introduced between the first and the second transformation. Transgenic lines either expressing ChiC or WD, or co‐expressing both genes were challenged with Fusarium oxysporum f.sp. nicotianae (Fon). Assessment of in vitro plant survival in the presence of Fon showed that transgenic lines co‐expressing both genes had significantly enhanced protection against the fungus (infection indices 0·0–1.·2) compared with corresponding isogenic lines expressing either of the genes (infection indices 2·5–9·8). Whole‐plant infection indices in transgenic lines were significantly related (r = 0·93, P < 0·01) to the extent of root colonization of the host, which ranged from 2·1% to 11·3% in lines co‐expressing both genes, and from 16·8% to 37·7% in lines expressing just one of the genes (compared with 86·4% in non‐transformed controls). Leaf extracts of transgenic lines also inhibited mycelial growth of Fon in vitro and caused hyphal abnormalities.     

Nursery practice on seed germination and seedling growth of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> using beneficial microbial inoculants

Nursery practice using microbial inoculants was performed to find out the efficacy of the inoculants on seed germination and seedling growth of sissoo (Dalbergia sissoo Roxb.). Microbial inoculants or effective microorganisms (EM) are a mixture of many different beneficial microorganisms in a solution. The seedlings were grown in a mixture of sandy soil and cowdung (3:1) kept in polybags with pouring EM solution at different concentrations (0.1%, 0.5%, 1%, 2%, 5% and 10%) before and after a week of sowing the seeds. Seed germination rate and growth parameters of seedlings were measured, such as, shoot and root length, vigor index, fresh and dry weight of shoot and root and total biomass increment. The nodulation status influenced by EM was also observed along with the measurement of pigment contents in leaves. The highest germination rate (69%) was observed in 2% EM treatment, followed by 67% and 65% in 1% and 5% EM. The highest shoot length (33.2 cm) was in 2% EM, whereas highest root length (26.3 cm) was in 1% EM. Both fresh and dry weights from shoot and root, were maximum (4.16 g and 1.57 g; 2.12 g and 0.83 g respectively) in 2% EM and were significantly (p ≤ 0.05) different from control. Vigor index was highest (4071) in 2% EM, which was significantly (p ≤ 0.05) different from control. Total dry weight increment was highest in 2% EM treatment, followed by 1% and 5% concentrations of EM. Nodulation number was higher at very low (0.1%) concentration of EM solution but it normally decreased with the increase of EM concentrations. The contents of chlorophyll a, chlorophyll b and carotenoid were highest (60.11, 17.05 and 42.48 mg·L⁻¹ respectively) in 2% EM treatment and lowest (39.35, 13.55 and 27.29 mg·L⁻¹ respectively) in control treatment. Therefore, low concentration of EM (up to 2%) can be used for getting maximum seed germination rate and seedling development of Dalbergia sissoo Roxb.     

Autologous red cell agglutination assay for HIV-1 antibodies: simplified test with whole blood

An antibody detection procedure based on agglutination of autologous red cells has been developed for samples of whole blood. A nonagglutinating monoclonal antibody to human red blood cells conjugated to a synthetic peptide antigen (in this case residues 579 to 601 of the HIV-1 envelope precursor, Arg-Ile-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp- Gly-Cys - Ser-Gly-Lys) permitted the detection of antibodies to the human immunodeficiency virus type 1 (HIV-1) in 10 microliters of whole blood within 2 minutes. Agglutination was specifically inhibited by addition of synthetic peptide antigen but not by unrelated peptides. The frequency of false positive results was 0.1% with HIV-1 seronegative blood donors (n = 874). The false negative results were approximately 1% (n = 81). The autologous red cell agglutination test is potentially suitable for simple, rapid, qualitative screening for antibodies to a variety of antigens of medical and veterinary diagnostic significance.     

Static and initiator protein-enhanced bending of DNA at a replication origin

DNA bending has been suggested to play a role in the regulation of gene expression, initiation of DNA replication, DNA packaging, and the recognition of specific DNA sequences by proteins. It has recently been demonstrated that DNA bending can be sequence-directed. Bent DNA has also been observed as a consequence of sequence-specific binding of proteins to DNA. In this report DNA of plasmid pT181 is shown to contain a bend at the replication origin. Furthermore, this bend is enhanced by the binding of the pT181 replication initiator protein, RepC, to the origin.     

Extinction risk assessment of the world’s seagrass species

Seagrasses, a functional group of marine flowering plants rooted in the world’s coastal oceans, support marine food webs and provide essential habitat for many coastal species, playing a critical role in the equilibrium of coastal ecosystems and human livelihoods. For the first time, the probability of extinction is determined for the world’s seagrass species under the Categories and Criteria of the International Union for the Conservation of Nature (IUCN) Red List of Threatened Species. Several studies have indicated that seagrass habitat is declining worldwide. Our focus is to determine the risk of extinction for individual seagrass species, a 4-year process involving seagrass experts internationally, compilation of data on species’ status, populations, and distribution, and review of the biology and ecology of each of the world’s seagrass species. Ten seagrass species are at elevated risk of extinction (14% of all seagrass species), with three species qualifying as Endangered. Seagrass species loss and degradation of seagrass biodiversity will have serious repercussions for marine biodiversity and the human populations that depend upon the resources and ecosystem services that seagrasses provide.