Enzymatic scarification of Anacamptis morio (Orchidaceae) seed facilitates lignin degradation, water uptake and germination.

The seed coat of many species contains hydrophobic lignins, and in soil the action of microbial ligninases may contribute to release from dormancy. Laboratory use of ligninases to stimulate germination is promising because of the specific action on the seed coat, whereas chemical scarification agents may also corrode the embryo. We hypothesised that exposure of Anacamptis morio (Orchidaceae) seeds to fungal laccase would stimulate germination, and that the mechanism involves lignin degradation and increased imbibition. Germination capacity in vitro was quantified with 1 U filter-sterilised laccase added to agar medium following autoclaving, compared to a 10% bleach solution (standard bleach surface sterilisation/scarification method used in orchid seed sowing). Lignin degradation was quantified using an optical method (phloroglucinol-HCl staining) combined with image analysis, following experimental pre-treatments involving immersion in laccase solution, distilled water (negative control) or bleach (positive control). Water uptake after experimental treatments was quantified as the proportion of seeds exhibiting visible uptake of an aqueous fluorochrome under UV excitation. Laccase stimulated a doubling of germination in vitro with respect to bleach surface sterilisation/scarification alone, from 23.7 to 49.8% (P = 0.007). Laccase and bleach methods both significantly decreased the optical signal of phloroglucinol (for laccase, to 79.9 ± 1.3% of controls; anova: F = 10.333, P = 0.002). Laccase resulted in a modest but highly significant (P < 0.0001) increase in water uptake with respect to the control (11.7%; cf 99.4% for bleach). Laccase scarification can stimulate germination of A. morio through a mechanism of targeted seed coat degradation. The results demonstrate the potential of this relatively non-invasive enzymatic scarification technique.

Complete sequencing and characterization of equine aggrecan.

OBJECTIVES: To fully sequence and characterize equine aggrecan and confirm conservation of major aggrecanase, calpain and matrix metalloproteinase (MMP) cleavage sites. METHODS: Reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends were used to generate clones that encompassed the complete equine aggrecan sequence. Clones were sequenced and compared with the equine genome database to determine intron-exon boundaries. RESULTS: The aggrecan gene spans over 61 kb on chromosome 1 and is encoded by 17 exons. Two major variants of aggrecan were cloned; one containing 8187 bp (2728 amino acids) and a second sequence of 8061 nucleotides (2686 amino acids). The variation was due to a CS1 domain polymorphism. Both sequences are substantially larger than predicted by the genomic database; 11 CS1 repeat elements are absent in the database sequence. The equine amino acid sequence was compared with human, bovine and murine sequences. Globular domains 1, 2 and 3 are highly conserved (overall identity over 80%). Equine CS1 is considerably larger than in other species and, therefore, is the least conserved domain (an overall amino acid identity of 22%). Previously defined aggrecanase, calpain and MMP cleavage sites were identified. Western blotting of chondrocyte culture samples showed complex post-secretion processing. CLINICAL SIGNIFICANCE: The complete equine aggrecan sequence will support more in-depth research on aggrecan processing and degradation in equine articular cartilage and other musculoskeletal tissues.

Phenotypic maintenance of articular chondrocytes in vitro requires BMP activity.

Articular chondrocytes are phenotypically unique cells that are responsible for the maintenance of articular cartilage. The articular chondrocytic phenotype is influenced by a range of soluble factors. In particular, members of the bone morphogenetic protein (BMP) family support the articular chondrocytic phenotype and stimulate synthesis of cartilaginous matrix. This study was carried out to determine the importance of BMPs in supporting the differentiated phenotype of articular chondrocytes in vitro. Exogenous BMP-2 supported expression of collagen type II and aggrecan in monolayer chondrocyte cultures, slowing the dedifferentiation process that occurs under these conditions. In contrast, BMP-2 had little effect on expression of these genes in three-dimensional aggregate cultures. Endogenous BMP-2 expression was lost in monolayer cultures, coincident with the down-regulation of collagen type II and aggrecan mRNAs, whereas BMP-2 mRNA levels were stable in aggregate cultures. Antagonism of endogenous BMP activity in aggregate cultures by Noggin or a soluble form of the BMP receptor resulted in reduced expression of collagen type II and aggrecan mRNAs, reduced collagen type II protein and sulfated glycosaminoglycan (GAG) deposition into the aggregate matrices and reduced secretion of GAGs into the culture media. These results indicate that endogenous BMPs are required for maintenance of the differentiated articular chondrocytic phenotype in vitro. These findings are of importance to cell-based strategies designed to repair articular cartilage. Articular chondrocytes require conditions that will support endogenous expression of BMPs to maintain the specialized phenotype of these cells.

Temporomandibular joint ankylosis and salivary mucocele in a cat: case report

An approximately 1.5-year-old, not neutered mixed breed cat was referred for evaluation of inability to open its mouth, and soft tissue swelling around the intermandibular region. Both signs were present since the cat was adopted, 1 year before presentation. The cause of the signs was not determined prior to referral. Based on the physical and radiographic examinations, left temporomandibular joint ankylosis and salivary mucocele were diagnosed. The lateral aspect of the condyloid process of the left mandible was removed and the salivary mucocele was treated by right mandibular and sublingual gland resection and drainage of the mucocele. After surgery, the cat showed good functional use of the mandible without discomfort.

Relata-se o caso de um gato de aproximadamente um ano e meio de idade, macho, não castrado, que foi encaminhado por apresentar incapacidade de abrir a boca e aumento de volume flutuante na região intermandibular. As lesões estavam presentes há um ano, desde quando o gato foi encontrado e adotado. A causa não foi determinada. Baseado nos exames físicos e radiográficos diagnosticou-se anquilose da articulação temporomandibular esquerda e mucocele salivar. O aspecto lateral do processo condilar da mandíbula esquerda foi removido, e a mucocele foi tratada por ressecção das glândulas salivares mandibular e sublingual direita e por drenagem da mucocele. Após a cirurgia, o gato mostrou bom uso funcional da mandíbula, sem sinais de desconforto.

Estudo comparativo entre flunizina meglumina ou buprenorfina no procedimento de ovariohisterectomia em gatas

O artigo não apresenta resumo.

Effect of fibrin glue derived from snake venom on the viability of autogenous split-thickness skin graft

The aim of this study was to analyze the effect of snake venom derived from fibrin glue on the viability of split-thickness skin graft. Nine crossbreed dogs were used. Full-thickness skin segments measuring 4X4 cm were bilaterally excised from the proximal radial area on each dog. A split-thickness skin graft was harvested from left lateral thoracic area using a freehand graft knife, and was secured to the left recipient bed using several simple interrupted sutures of 3-o nylon (sutured graft). A split-thickness skin graft was harvested from the right lateral thoracic area using a graft knife. Fibrin glue derived from snake venom was applied to the recipient bed, and 8 equidistant simple interrupted sutures of 3-0 nylon were used to secure the skin graft (glued graft). Viable and nonviable areas were traced on a transparent sheet and measured using a Nikon Photomicroscope connected to a KS-300 image analysis system. The skin graft and recipient bed were collected from three dogs at day 7,15, and 30 postoperative. The glued grafs had statistically higher graft viability than sutured grafts. Histological examination showed that the tissue repair process in the glued grafts was more accentuated than sutured grafts. It was possible to conclude that fibrin glue derived from snake venom increased survival of autogenous split-thickness skin graft.

A georeferenced river quality model.

Water quality models have reached a high degree of sophistication, but their weak side remains user interface and output georeferencing. The aim of this paper is to propose an interfacing procedure between two widespread but specialised programming environments: ArcVIEW as a Geographical Information System (GIS) and Matlab as a scientific programming tool for numerical analysis. The proposed solution is based on a Dynamic Data Exchange (DDE) between the two programs in order to operate a Matlab-based water quality model from within the GIS environment. It is shown how special GIS objects must be created and how they operate to achieve the goal of having quality data created by the model placed on a geographical map, together with other site features.

Linkage Arrangement of RFLP loci in progenies from crosses between doubled haploid Asparagus officinalis L. clones.

A preliminary genetic map of the dioecious species Asparagus officinalis L. (2n = 20) has been constructed on the basis of restriction fragment length polymorphism (RFLP) and isozyme marker data. With DNA samples digested with either EcoRI or HindIII 61 out of 148 probes (41%) identified RFLPs in six families of doubled haploid lines obtained through anther culture. A higher level of polymorphism (65%) was observed when a single family was screened for RFLPs using six distinct restriction enzymes. Segregation analysis of the BC progenies (40-80 individuals) resulted in a 418-cM extended map comprising 43 markers: 39 RFLPs, three isozymes and one morphological (sex). These markers are clustered in 12 linkage groups and four of them exhibited significant deviations from the expected 1∶1 ratio. One isozyme and three RFLP markers were assigned to the sex chromosome.