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A nine-year-old female desexed Great Dane presented with a painful, proliferative, soft red putative neoplastic vascular mass on the nictitating membrane. Three 7-day cycles of the topical cytotoxic drug mitomycin C 0.04%, applied four times daily to the lesion using a low-dose alternate-week pulse therapy, brought about rapid remission of the lesion. The lesion was still in remission at time of euthanasia some 13 months later.

Background and purpose P2X4 receptors are emerging therapeutic targets for treating chronic pain and cardiovascular disease. Dogs are well-recognised natural models of human disease, but information regarding P2X4 receptors in dogs is lacking. To aid the development and validation of P2X4 receptor ligands, we have characterised and compared canine and human P2X4 receptors. Experimental approach Genomic DNA was extracted from whole blood samples from 101 randomly selected dogs and sequenced across the P2RX4 gene to identify potential missense variants. Recombinant canine and human P2X4 receptors tagged with Emerald GFP were expressed in 1321N1 and HEK293 cells and analysed by immunoblotting and confocal microscopy. In these cells, receptor pharmacology was characterised using nucleotide-induced Fura-2 AM measurements of intracellular Ca2+ and known P2X4 receptor antagonists. P2X4 receptor-mediated inward currents in HEK293 cells were assessed by automated patch clamp. Key results No P2RX4 missense variants were identified in any canine samples. Canine and human P2X4 receptors were localised primarily to lysosomal compartments. ATP was the primary agonist of canine P2X4 receptors with near identical efficacy and potency at human receptors. 2'(3')-O-(4-benzoylbenzoyl)-ATP, but not ADP, was a partial agonist with reduced potency for canine P2X4 receptors compared to the human orthologues. Five antagonists inhibited canine P2X4 receptors, with 1-(2,6-dibromo-4-isopropyl-phenyl)-3-(3-pyridyl)urea displaying reduced sensitivity and potency at canine P2X4 receptors. Conclusion and implications P2X4 receptors are highly conserved across dog pedigrees and display expression patterns and pharmacological profiles similar to human receptors, supporting validation and use of therapeutic agents for P2X4 receptor-related disease onset and management in dogs and humans.

Caudal duplication (dipygus) is an uncommon pathologic of conjoined twinning. The conjoined malformation is classified according to the nature and site of the union. We report the presence of this malformation in a female crossbreed puppy. The puppy was delivered by caesarean section following a prolonged period of dystocia. External findings showed a single head (monocephalus) and a normal cranium with no fissure in the medial line detected. The thorax displayed a caudal duplication arising from the lumbosacral region (rachipagus). The puppy had three upper limbs, a right and left, and a third limb in the dorsal region where the bifurcation began. The subsequent caudal duplication appeared symmetrical. Necropsy revealed internal abnormalities consisting of a complete duplication of the urogenital system and a duplication of the large intestines arising from a bifurcation of the caudal ileum. Considering the morphophysiological description the malformation described would be classified as the first case in the dog of a monocephalus rachipagus tribrachius tetrapus.

Case report A 2-year-old male Airedale Terrier was presented with an acute and painful, predominantly ventral, vesicular skin eruption following putative dermal contact with Plumbago auriculata (Sky Flower, Leadwort). Prompt dermal decontamination and supportive therapy brought about a rapid recovery in the patient. Conclusion Contact with botanical triggers is an important consideration for causes of acute vesicular skin conditions in dogs.

Protein kinase B (PKB or Akt) is an important component of intracellular signaling pathways regulating growth and survival. Signaling through PKB is frequently deregulated in cancer, and inhibitors of PKB therefore have potential as antitumor agents. The optimization of lipophilic substitution within a series of 4-benzyl-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidin-4-amines provided ATP-competitive, nanomolar inhibitors with up to 150-fold selectivity for inhibition of PKB over the closely related kinase PKA. Although active in cellular assays, compounds containing 4-amino-4-benzylpiperidines underwent metabolism in vivo, leading to rapid clearance and low oral bioavailability. Variation of the linker group between the piperidine and the lipophilic substituent identified 4-amino-1-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperidine-4-carboxamides as potent and orally bioavailable inhibitors of PKB. Representative compounds modulated biomarkers of signaling through PKB in vivo and strongly inhibited the growth of human tumor xenografts in nude mice at well-tolerated doses.

Objective To determine the thyroid status of Basenji dogs in Australia. Methods Jugular or cephalic venipuncture blood samples were taken from 113 Basenji, comprising 47 males, 5 castrates, 48 entire and 13 spayed bitches, and sent on ice in plain and EDTA tubes to a single laboratory to determine haematocrit and serum concentrations of total thyroid hormone (thyroxine, TT4), thyroid-stimulating hormone (TSH) and cholesterol. In a subgroup of 8 dogs with abnormal elevated TSH concentrations and subnormal TT4 concentrations, 5 were further examined by dynamic endocrine testing using recombinant human (rh) TSH (54 µg). Results Ages ranged from 1 to 14 years and weight range was 6.5 to 14.0 kg. TT4 concentrations (nmol/L) ranged from 2 to 27, with a median of 13 and a mean ± SD of 13.0 ± 5.7. Importantly, 85/113 (75%) of TT4 values were lower than the normal laboratory reference range (17–37). TSH concentrations (ng/mL) ranged from 0.05 to 5.37, with a median of 0.16 and a mean ± SD of 0.3 ± 0.6. Conclusions Basenji have a similar reference range for serum TSH, but a considerably lower reference range for TT4 (2–27 nmol/L) than most breeds and crossbreds, resembling the sight hounds in this respect. Given the difficulty of accurately measuring TT4 concentrations that are so low, concomitant serial TSH determinations are essential to properly asses thyroid function. Taken alone, TT4 determinations are only of use when the value is within the reference range, in which case a diagnosis of hypothyroidism is likely excluded.

The application of fragment-based screening techniques to cyclin dependent kinase 2 (CDK2) identified multiple (>30) efficient, synthetically tractable small molecule hits for further optimization. Structure-based design approaches led to the identification of multiple lead series, which retained the key interactions of the initial binding fragments and additionally explored other areas of the ATP binding site. The majority of this paper details the structure-guided optimization of indazole (6) using information gained from multiple ligand−CDK2 cocrystal structures. Identification of key binding features for this class of compounds resulted in a series of molecules with low nM affinity for CDK2. Optimisation of cellular activity and characterization of pharmacokinetic properties led to the identification of 33 (AT7519), which is currently being evaluated in clinical trials for the treatment of human cancers.

Redox changes are one of the factors that influence cell-cycle progression and that control the processes of cellular proliferation, differentiation, senescence and apoptosis. Proteins regulated through redox-sensitive cysteines have been characterized but specific ‘sulphydryl switches’ in replication proteins remain to be identified. In bovine papillomavirus type-1, DNA replication begins when the viral transcription factor E2 recruits the viral initiator protein E1 to the origin of DNA replication (ori). Here we show that a novel dimerization interface in the E2 transcription activation domain is stabilized by a disulphide bond. Oxidative cross-linking via Cys57 sequesters the interaction surface between E1 and E2, preventing pre-initiation and replication initiation complex formation. Our data demonstrate that as well as a mechanism for regulating DNA binding, redox reactions can control replication by modulating the tertiary structure of critical protein factors using a specific redox sensor.

The relative function of the P2X7 receptor, an ATP-gated ion channel, varies between humans due to polymorphisms in the P2RX7 gene. This study aimed to assess the functional impact of P2X7 variation in a random sample of the canine population. Blood and genomic DNA were obtained from 69 dogs selected as representatives of a cross section of different breeds. P2X7 function was determined by flow cytometric measurements of dye uptake and patch-clamp measurements of inward currents. P2X7 expression was determined by immunoblotting and immunocytochemistry. Sequencing was used to identify P2RX7 gene polymorphisms. P2X7 was cloned from an English springer spaniel, and point mutations were introduced into this receptor by site-directed mutagenesis. The relative function of P2X7 on monocytes varied between individual dogs. The canine P2RX7 gene encoded four missense polymorphisms: F103L and P452S, found in heterozygous and homozygous dosage, and R270C and R365Q, found only in heterozygous dosage. Moreover, R270C and R365Q were associated with the cocker spaniel and Labrador retriever, respectively. F103L, R270C, and R365Q but not P452S corresponded to decreased P2X7 function in monocytes but did not explain the majority of differences in P2X7 function between dogs, indicating that other factors contribute to this variability. Heterologous expression of site-directed mutants of P2X7 in human embryonic kidney-293 cells indicated that the R270C mutant was nonfunctional, the F103L and R365Q mutants had partly reduced function, and the P452S mutant functioned normally. Taken together, these data highlight that a R270C polymorphism has major functional impact on canine P2X7.

Two experiments were carried out, one in 1995 and one in 1997, to investigate the competitive abilities of two spring wheat cultivars with Sinapis arvensis L. The spring wheat cultivars (Baldus and Canon) of contrasting growth habit were grown with and without S. arvensis under two different moisture regimes (10% and 70% of field capacity). In 1995, S. arvensis was found to be less competitive when subjected to moisture stress, resulting in smaller wheat yield losses in dry soil than in moist soil. In both years, seed production of S. arvensis was reduced by competition and moisture stress, and the seeds produced by plants that had been grown in drier soil were small and had negligible dormancy. Hence, in dry conditions, the competitiveness of S. arvensis and its potential to produce persistent seed may be reduced. Some differences between the two wheat cultivars were evident: cv. Baldus was more competitive against S. arvensis than cv. Canon. This could be attributed to differences in canopy structure.

Weed suppression characteristics of different winter cereal species and cultivars were studied in two field experiments in two successive years. Two cultivars each of barley, oats and wheat were studied using the weed Galium aparine L. at a range of densities. Significant differences were found in the suppressive abilities of the crop species: oats being the most suppressive, followed by barley and then wheat. Percentage yield loss was related to G. aparine density using a rectangular hyperbolic model. In 1995-96, significant differences between the two wheat cultivars were found, with cv. Avalon being able to suppress G. aparine growth much more effectively than cv. Spark. However, in 1996-97, high rainfall in May, June and July enabled G. aparine to escape suppression by the cultivars. Cultivar competitive ability was associated with high overall leaf area, resistance to loss of tillers under competitive pressure, greater height, canopy structure and development. The oat cultivars were more competitive at early growth stages than could be accounted for by their canopy structure; the possible contribution of allelopathic exudates to their suppressive ability is discussed.