Orbital floor reconstruction with poly-L/D-lactide implants: clinical, radiological and immunohistochemical study in sheep.

In this study the reconstruction capacity of orbital wall in sheep was evaluated when poly-L/D-lactide (PLDLA96) implants were used for large blow-out defects in 18 sheep. The contralateral side, where the defects healed spontaneously, served as controls. The follow-up was 12, 16, 22 and 36 weeks. Healing was evaluated clinically, radiologically, histologically and immunohistochemically. Physiochemical properties of the implants were also studied. At first, the implants were surrounded by elastic capsules, which gradually ossified. At 36 weeks, 60% were still visible and deformed but surrounded by bone. Light microscopy revealed a low grade inflammatory reaction. Expression of Tn-c and cFn was intense throughout the study. Shear strength decreased gradually and was not measurable after 16 weeks. Crystallinity increased steadily from 1.5 to 29.30% and molecular weight decreased from 49,000 to 4186. In CT, the final bony defect was smaller in the reconstructed sides than in the controls. Based on this study it can be concluded that PLDLA96 implant provokes a local inflammation, which does not prevent bone healing. The deformation of the implant, however, indicates that this PLDLA96 plate is not suitable for orbital floor reconstruction.

Cloning and analysis of the cDNA encoding the rod G-protein transduction alpha, beta1 and gamma1 subunits from the canine retina.

The canine (Canis familiaris) retinal rod transducin (G(T)) alpha, beta1 and gamma1 subunits were sequenced. Cloning of the cDNAs was accomplished by polymerase chain reaction (PCR) using degenerate and wild type retinal cDNA libraries as templates. The deduced amino acid sequences were highly similar to rod transducins from other species: G(T alpha) differed by 5 amino acids from the corresponding human sequence, whereas beta1 and gamma1 were identical to human sequences. The coding sequence of rod transducin was evaluated as a possible cause for the recessively inherited retinal rod-cone degeneration: there were no nucleotide differences between the wild type and retinal degenerate strains.

Cloning of the cDNA encoding rod photoreceptor cGMP-phosphodiesterase alpha and gamma subunits from the retinal degenerate Labrador retriever dog.

The nucleotide (nt) sequence of the cDNA encoding the retinal rod cyclic 3'5'-GMP phosphodiesterase (PDE) alpha and gamma subunits from two strains of dogs-(i) Labrador Retrievers homozygous for autosomally recessively inherited rod-cone degeneration and (ii) the wild-type Beagle-are reported. Cloning of these subunits was accomplished by polymerase chain reaction using retinal cDNA libraries as templates. The nt sequence of alpha PDE predicts a 861-amino-acid polypeptide which is 97.7 per cent and 96.9 per cent identical to the bovine and human counterparts, respectively. PDE gamma encodes an 87-amino-acid polypeptide differing from bovine and murine gamma subunits by only one amino acid. Since no differences were found between these two strains of dogs, the cause of the Labrador Retriever's degeneration remains to be determined.

Impaired retinal function in young labrador retriever dogs heterozygous for late onset rod-cone degeneration.

Xenon-flash d.c.-electroretinograms were recorded from dark adapted, rod-cone degenerate homozygote affected (n = 6), heterozygote carrier (n = 3) and control retinas (n = 4) at 3 and 4 months of age, starting at 0.6 log units below control PII threshold. One log unit higher stimuli were necessary to evoke PII in heterozygote and affected retinas compared to controls. Unique to the heterozygotes, double peaked PII responses that were evoked by -2 log relative units intensity stimulation were significantly (P = 0.028) lower in amplitude than those of controls. PII amplitudes of homozygotes were significantly (P = 0.005) lower in amplitude than those of controls at both ages examined in response to -2 and 0 log relative intensity stimulation. No differences were found in scotopic threshold response amplitudes or times to peak between the three groups. Homozygote affected PII times to peak were significantly (P = 0.005) shorter in relation to controls at -2 log units. Findings suggest that heterozygotes exhibit an impaired retinal function which can be demonstrated at 3 and 4 months in this mutant.

The introns of the canine rod opsin gene show higher sequence homology to the human than to the rodent introns.

Using genomic DNA from late-onset retinal degenerate and wild type Labrador Retrievers as templates and canine exon-specific oligonucleotides as primers in polymerase chain reaction, all four introns of opsin were cloned and sequenced. Dot-matrix comparisons were made for human, murine and canine introns. Selected sequences containing either intronic or coding sequences were aligned and used for phylogenetic relationship analysis. The opsin gene introns are conserved between the human, the mouse and the dog with regards to number and length. In addition there is an astonishingly high degree of sequence homology between the second and fourth introns. Introns 2(1277 bp in dog) and 4 (863 bp in dog) are 72% and 71% homologous to the human introns, and 57% and 52% homologous to the mouse introns, respectively. The coding sequence (CDS) of the dog shows 93% homology to human CDS and 88% homology to mouse CDS. A phylogenetic analysis of the intronic sequences 2 and 4 confirms the higher relatedness between dog and human than between mouse and human opsin genes. As there are good reasons to believe that the primate and rodent lineages are closer to each other than to the Canis familiaris, there must be some functional constraints on the evolution of human and dog opsins.

Propofol influences the electroretinogram to a lesser degree than thiopentone.

BACKGROUND: The Electroretinogram (ERG) is used clinically to assess the function of retina. Anaesthetic agents are known to affect ERG, and as anaesthesia is often needed in children and uncooperative patients, knowledge about its effects is of clinical importance. Barbiturates selectively depress ERG components, and we compared thiopentone with propofol to assess if the latter preserved retinal function better. METHODS: Ten pigs, average weight 17 kg (SD +/- 2 kg) were anaesthetized randomly with propofol 10 mg kg-1 or thiopentone 30 mg kg-1. Anaesthesia was maintained by 65% nitrous oxide in oxygen and continuous infusion of the induction agent, i.e. 10 mg kg-1 h-1 of propofol, or 10 mg kg-1 h-1 for the first hour, then 5 mg kg-1 h-1 of thiopentone, with doses being based on pilot studies. After an interval of one week the programme was repeated using the other agent. After 40 minutes dark-adaptation, responses to single flashes of graded intensities from a xenon flashlamp were recorded at five-minute intervals. The a- and b-wave amplitudes and implicit times (time to peak), and a-wave slopes were determined. RESULTS: The b-wave implicit time was significantly shorter during propofol anaesthesia than when using thiopentone. The effect was most pronounced at the lowest intensities (P < 0.01). No statistically significant differences were found in the amplitudes of the b-waves. The a-wave appeared at lower stimulus intensity (P < 0.05) and the a-wave slopes were significantly steeper (P < 0.01) during propofol anaesthesia. CONCLUSION: Propofol accordingly appeared to preserve the photoreceptor response better than thiopentone, and may therefore be considered to be more suitable for ERG recordings than thiopentone.

Elevation of cGMP with normal expression and activity of rod cGMP-PDE in photoreceptor degenerate labrador retrievers.

Cyclic guanosine 3',5'-monophosphate (cGMP) levels were determined in retinas from a strain of Labrador Retrievers with inherited retinal dystrophy manifesting at early stages of retinal differentiation. The cGMP contents of dystrophic retinas of dogs from 1 to 4 months of age (n = 7) were significantly higher (p = 0.001) than in age-matched controls of the same breed (n = 11). Ultrastructure along the vertical retinal meridian was studied in developing retinas and findings were related to those of age-matched wild-type controls of the same breed. Slow central to peripheral progression of degeneration was observed in affected dogs. No differences were found in total cGMP-phosphodiesterase (PDE) activity, in PDE subunit composition as determined by Western blotting of 2-month-old homozygote affected retinas, or in the amino acid sequence deduced from the nucleotide sequence of the PDE beta-subunit as compared to controls. This model of photoreceptor degeneration thus is the first case of an apparent abnormality of cGMP metabolism that is not associated with a defect in the PDE catalytic subunits, and it is also the first reported model not associated with severe developmental abnormalities and rapid degeneration.

Early morphometry of a retinal dystrophy in Labrador retrievers.

Retinal morphometry was assessed in 7 dogs from a colony of Labrador Retrievers with dystrophic retinas at 1,2,3,4 and 18 months of age. Rod outer segment length and outer nuclear layer width were measured in the central, midperipheral and peripheral retina at six locations along the vertical meridian. Early striking regional differences in onset and rate of progression were characteristic for this inherited retinal degeneration. Notably, some areas of the retina developed fully and normally before degenerating. The central parts of the vertical meridians showed slightly disorganized rod outer segments already at 1 month of age and they were significantly shorter than those of control animals at 3 and 4 months (p < 0.01 and p < 0.001, respectively). The rod outer segments of the midperipheral and peripheral regions were, however, comparable to control animals as late as at 4 months of age. At 18 months the rod outer segments of dystrophic animals were significantly shorter in all retinal regions (p < 0.0005). At the age the outer nuclear layer of the dystrophic animals had become significantly thinner than that of control animals in all retinal regions (p < 0.001), indicating a clear visual cell loss. It is reasonable to characterize this as a retinal degeneration having a relatively slow progression, which enhances its relevance to conditions of clinical significance.