The INK4a-ARF (CDKN2A)-locus on chromosome 9p21 encodes for two tumour suppressor proteins, p16 INK4a and p14 ARF , that act as upstream regulators of the Rb-CDK4 and p53 pathways. To study the contribution of each pathway in tumorigenesis of hepatocellular carcinoma (HCC), we analysed the alterations of p14 ARF , p16 INC4a and p53. After microdissection, DNA of 71 hepatocellular carcinomas was analysed for INK4-ARF inactivation and p53 mutation by DNA sequence analysis, methylation-speci®c PCR (MSP), restriction-enzyme related polymerase chain reaction (RE ± PCR), mRNA expression and immunohistochemistry. In addition, microdeletion of p14 ARF and p16 INC4a were assessed by di erential PCR. Inactivation of p14 ARF was found in 11/71 cases (15%), alterations of p16 INK4a occurred in 47/71 carcinomas (66%), which correlated with loss of mRNA transcription. Five tumours (7%) had homozygous deletions of the INK4a-ARF locus. We failed to detect speci®c mutations of both exons. P16 INK4a methylation with an unmethylated p14 ARF promotor appeared in 39 cases. Mutations of p53 were found in 30 of 71 HCC (42%), and only one of them harboured p14 ARF inactivation. We failed to establish alterations of the INK4a-ARF locus or p53 status as independent prognostic factor in these tumours. Our data indicate, that p14 ARF methylation occurs independently of p16 INK4a alterations in a subset of HCC together with wild type p53. The INK4a-ARF-/p53-pathway was disrupted in 86% of HCC, either by p53 mutations or by INK4a-ARF inactivation, and may have co-operative e ects in hepatocarcinogenesis. Oncogene (2001) 20, 7104 ± 7109.

Ophthalmic complications are common in patients with retrobulbar inflammation indicating that these patients should undergo ophthalmic assessment and follow-up.

Synapses represent specialized cell-cell contact sites between nerve cells. These structures mediate the rapid and efficient transmission of signals between neurons and are surrounded by glial cells. Previous investigations have shown that astrocytes are important for the formation, maintenance, and function of CNS synapses. To study effects of glial-derived molecules on synaptogenesis, we have established an in vitro cell-insert coculture system for E18 rat hippocampal neurons and various glial cell types. Neurons were cultured without direct contact with glial cells for distinct time periods. First, it was confirmed that astrocytes are essential to promote survival of E18 hippocampal neurons. Beginning with 10 days in culture, the concurrent expression of pre- and postsynaptic proteins was observed. Moreover, the colocalization of the presynaptic marker Bassoon and the postsynaptic protein ProSAP1/Shank2 indicated the formation of synapses. A technique was developed that permits the semiautomated quantitative determination of the number of synaptic puncta per neuron. The culture system was used to assess effects of pharmacological treatments on synapse formation by applying blockers and activators of small GTPases. In particular, treatment with lysophosphatidic acid enhanced synaptogenesis in the coculture system.

A 6-year-old, female, spayed Border Collie was presented to the Unit of Comparative Ophthalmology at the Animal Health Trust with a 6-month history of a progressive nonpainful opacity of the left cornea. A keratectomy was performed and the tissue submitted for histopathology. The diagnosis was squamous cell carcinoma. There has been no recurrence of the neoplasm to date (5 months). Canine corneal squamous cell carcinoma (SCC) has not been reported previously in the UK.

BackgroundSulfated glycosaminoglycan chains are known for their regulatory functions during neural development and regeneration. However, it is still unknown whether the sulfate residues alone influence, for example, neural precursor cell behavior or whether they act in concert with the sugar backbone. Here, we provide evidence that the unique 473HD-epitope, a representative chondroitin sulfate, is expressed by spinal cord neural precursor cells in vivo and in vitro, suggesting a potential function of sulfated glycosaminoglycans for spinal cord development.ResultsThus, we applied the widely used sulfation inhibitor sodium chlorate to analyze the importance of normal sulfation levels for spinal cord neural precursor cell biology in vitro. Addition of sodium chlorate to spinal cord neural precursor cell cultures affected cell cycle progression accompanied by changed extracellular signal-regulated kinase 1 or 2 activation levels. This resulted in a higher percentage of neurons already under proliferative conditions. In contrast, the relative number of glial cells was largely unaffected. Strikingly, both morphological and electrophysiological characterization of neural precursor cell-derived neurons demonstrated an attenuated neuronal maturation in the presence of sodium chlorate, including a disturbed neuronal polarization.ConclusionsIn summary, our data suggest that sulfation is an important regulator of both neural precursor cell proliferation and maturation of the neural precursor cell progeny in the developing mouse spinal cord.

Objectives To describe the use of corneal autografts for repair of deep corneal defects in dogs. Materials and Methods Retrospective analysis of clinical records of dogs that received autologous corneal grafts. Results Fifteen dogs (16 eyes) of different breed, age and gender were included. Brachycephalic breeds were overrepresented (10/15 dogs). Defects were unilateral in 14 dogs and bilateral in one dog, extended to at least 80% of the stromal thickness in all eyes, with descemetoceles in four eyes and corneal perforations in five eyes. Most ulcers (13/16 eyes) were located centrally. Corneal autografts were harvested from healthy peripheral cornea of the ipsilateral eye. The thickness of the autograft was limited to a set depth of 0.3 mm. The autograft was sutured into a previously debrided ulcer bed with a continuous or simple interrupted suture pattern using absorbable or non‐absorbable suture material. Additional interventions included a partial temporary tarsorrhaphy and bandage contact lenses. Postoperatively patients received topical antibiotics and systemic anti‐inflammatory drugs, and 12/15 dogs also received systemic antibiotics. Mean follow‐up time was 54 days (2 to 462). In all eyes the donor site healed uneventfully with mild, persistent corneal fibrosis. Postoperative complications included autograft keratomalacia, graft dehiscence and corneal pigmentation. No patient required additional surgery. Good structural and functional outcome was accomplished in 14 of 16 eyes. Clinical significance Autologous corneal grafts provide tectonic support and result in good corneal transparency in selected cases of dogs with deep to perforated corneal ulcerations.

Background Bilateral perisylvian polymicrogyria (BPP) is a well-recognized malformation of cortical development commonly associated with epilepsy, cognitive impairment, and oromotor apraxia. Reports have suggested the association of BPP with arthrogryposis multiplex congenita. We sought to investigate the clinical, electrophysiological, and neuroradiological features of this combined syndrome to determine if there are unique features that distinguish BPP with arthrogryposis from BPP alone. Methods Cases of BPP with congenital arthrogryposis were identified from a large research database of individuals with polymicrogyria. Clinical features (including oromotor function, seizures, and joint contractures), MR brain imaging, and results of neuromuscular testing were reviewed. Results Ten cases of BPP with congenital arthrogryposis were identified. Most cases had some degree of oromotor apraxia. Only a few had seizures, but a majority of cases were still young children. Electrophysiological studies provided evidence for lower motor neuron or peripheral nervous system involvement. On brain imaging, bilateral polymicrogyria (PMG) centered along the Sylvian fissures was seen, with variable extension frontally or parietally; no other cortical malformations were present. We did not identify obvious neuroimaging features that distinguish this syndrome from that of BPP without arthrogryposis. Conclusions The clinical and neuroimaging features of the syndrome of BPP with congenital arthrogryposis appear similar to those seen in cases of isolated BPP without joint contractures, but electrophysiological studies often demonstrate coexistent lower motor neuron or peripheral nervous system pathology. These findings suggest that BPP with arthrogryposis may have a genetic etiology with effects at two levels of the neuraxis.

Objectives: To describe MRI features of canine retrobulbar inflammation, their association with clinical findings and outcome and to assess the value of MRI in detecting orbital foreign bodies. Materials and MethOds:Clinical records of dogs with confirmed (23 of 31) and suspected (eight of 31) retrobulbar inflammation that underwent low field MRI studies were analysed retrospectively.results: Of the 31 dogs included in the study there was abscessation in 19. Extraocular myositis (27 of 31) was concurrent with strabismus in three cases. Of 25 patients with exophthalmos, 14 had concurrent optic nerve swelling and, of these, five had permanent loss of vision. There was no vision loss in patients without nerve swelling. One case of suspected retinal detachment on MRI was confirmed clinically. Extensive abnormal contrast enhancement in the temporalis, masseter and pterygoideus muscles was associated with facial (n=3) and trigeminal nerve deficits (n=1). Three patients with inflammation extending into the nasal cavity and frontal sinus (one of 31) or meningeal contrast enhancement (two of 31), showed optic and oculomotor nerve deficits. On MRI a foreign body was not visible in 20 of 31 case or "appeared likely" in 11 of 31 dogs. A foreign body was found at surgery in one case. clinical significance: MRI outlines the extent of retrobulbar inflammation. Clinical findings were associated with imaging findings. MRI overestimated the presence of foreign bodies. ttp://www.bsava.com/ M. C. Fischer et al.