Correlation between Multiparametric MR Imaging and Molecular Genetics in Pontine Pediatric High-Grade Glioma.

BACKGROUND AND PURPOSE: Molecular profiling is a crucial feature in the "integrated diagnosis" of CNS tumors. We aimed to determine whether radiomics could distinguish molecular types of pontine pediatric high-grade gliomas that have similar/overlapping phenotypes on conventional anatomic MR images. MATERIALS AND METHODS: Baseline MR images from children with pontine pediatric high-grade gliomas were analyzed. Retrospective imaging studies included standard precontrast and postcontrast sequences and DTI. Imaging analyses included median, mean, mode, skewness, and kurtosis of the ADC histogram of the tumor volume based on T2 FLAIR and enhancement at baseline. Histone H3 mutations were identified through immunohistochemistry and/or Sanger or next-generation DNA sequencing. The log-rank test identified imaging factors prognostic of survival from the time of diagnosis. Wilcoxon rank-sum and Fisher exact tests compared imaging predictors among groups. RESULTS: Eighty-three patients had pretreatment MR imaging and evaluable tissue sampling. The median age was 6 years (range, 0.7-17 years); 50 tumors had a K27M mutation in H3-3A, and 11, in H3C2/3. Seven tumors had histone H3 K27 alteration, but the specific gene was unknown. Fifteen were H3 wild-type. Overall survival was significantly higher in H3C2/3- compared with H3-3A-mutant tumors (P = .003) and in wild-type tumors compared with any histone mutation (P = .001). Lower overall survival was observed in patients with enhancing tumors (P = .02) compared with those without enhancement. H3C2/3-mutant tumors showed higher mean, median, and mode ADC_total values (P < .001) and ADC_enhancement (P < .004), with lower ADC_total skewness and kurtosis (P < .003) relative to H3-3A-mutant tumors. CONCLUSIONS: ADC histogram parameters are correlated with histone H3 mutation status in pontine pediatric high-grade glioma.

Invited Review: Emerging functions of histone H3 mutations in paediatric diffuse high-grade gliomas.

Paediatric diffuse high-grade gliomas (pHGG) are rare, but deadly tumours. The discovery of recurrent mutations in the tail of histone H3, changing lysine 27 to methionine, or glycine 34 to arginine or valine, has illuminated a critical role for epigenetic dysregulation in the aetiology of childhood gliomas and opened new avenues of exploration that have resulted in numerous advances for the field. In this review, we describe the current models of H3K27M mutant cancer that are available to the research community and the insights they have provided on tumour biology and the epigenetic and transcriptional effects of histone mutations. We also review the current understanding of the H3G34R/V mutation and the therapeutic outlook for the treatment of pHGG.

Dual inhibition of CDK4/Rb and PI3K/AKT/mTOR pathways by ON123300 induces synthetic lethality in mantle cell lymphomas.

This study describes the characterization of a novel kinase inhibitor, ON123300, which inhibits CDK4/6 (cyclin-dependent kinases 4 and 6) and phosphatidylinositol 3 kinase-δ (PI3K-δ) and exhibits potent activity against mantle cell lymphomas (MCLs) both in vitro and in vivo. We examined the effects of PD0332991 and ON123300 on cell cycle progression, modulation of the retinoblastoma (Rb) and PI3K/AKT pathways, and the induction of apoptosis in MCL cell lines and patient-derived samples. When Granta 519 and Z138C cells were incubated with PD0332991 and ON123300, both compounds were equally efficient in their ability to inhibit the phosphorylation of Rb family proteins. However, only ON123300 inhibited the phosphorylation of proteins associated with the PI3K/AKT pathway. Cells treated with PD0332991 rapidly accumulated in the G0/G1 phase of cell cycle as a function of increasing concentration. Although ON123300-treated cells arrested similarly at lower concentrations, higher concentrations resulted in the induction of apoptosis, which was not observed in PD0332991-treated samples. Mouse xenograft assays also showed a strong inhibition of MCL tumor growth in ON123300-treated animals. Finally, treatment of ibrutinib-sensitive and -resistant patient-derived MCLs with ON123300 also triggered apoptosis and inhibition of the Rb and PI3K/AKT pathways, suggesting that this compound might be an effective agent in MCL, including ibrutinib-resistant forms of the disease.

Gabapentin-induced pharmacodynamic effects in the spinal nerve ligation model of neuropathic pain.

BACKGROUND: The function of brain networks can be changed in a maladaptive manner in response to chronic neuropathic pain. Analgesics can reduce pain by acting on such networks via direct or indirect (peripheral or spinal) mechanisms. This investigation aimed to map gabapentin's pharmacodynamics (PD) in the rodent brain following induction of neuropathic pain in order to further understand its PD profile. METHODS: Pharmacological magnetic resonance imaging (phMRI) and a novel functional connectivity analysis procedure were performed following vehicle or gabapentin treatment in the rat spinal nerve ligation (SNL) model of neuropathic pain as well as sham animals. RESULTS: phMRI performed in SNL animals revealed robust gabapentin-induced responses throughout the hippocampal formation, yet significant (p < 0.05, corrected for multiple comparisons) responses were also measured in other limbic structures and the sensorimotor system. In comparison, sham animals displayed weaker and less widespread phMRI signal changes subsequent to gabapentin treatment. Next, communities of networks possessing strong functional connectivity were elucidated in vehicle-treated SNL and sham animals. We observed that SNL and sham animals possessed distinct functional connectivity signatures. When measuring how gabapentin altered the behaviour of the discovered networks, a decrease in functional connectivity driven by gabapentin was not only observed, but the magnitude of this PD effect was greater in SNL animals. CONCLUSIONS: Using phMRI and functional connectivity analysis approaches, the PD effects of gabapentin in a preclinical neuropathic pain state were characterized. Furthermore, the current results offer insights on which brain systems gabapentin directly or indirectly acts upon.

Review: insights gained from modelling high-grade glioma in the mouse.

High-grade gliomas (HGGs) are devastating primary brain tumours with poor outcomes. Advances towards effective treatments require improved understanding of pathogenesis and relevant model systems for preclinical testing. Mouse models for HGG provide physiologically relevant experimental systems for analysis of HGG pathogenesis. There are advantages and disadvantages to the different methodologies used to generate such models, including implantation, genetic engineering or somatic gene transfer approaches. This review highlights how mouse models have provided insights into the contribution of specific mutations to tumour initiation, progression and phenotype, the influence of tumour micro-environment, and the analysis of cell types that can give rise to glioma. HGGs are a heterogeneous group of tumours, and the complexity of diverse mutations within common signalling pathways as well as the developmental and cell-type context of transformation contributes to the overall diversity of glioma phenotype. Enhanced understanding of the mutations and cell types giving rise to HGG, along with the ability to design increasingly complex mouse models that more closely simulate the process of human gliomagenesis will continue to provide improved experimental systems for dissecting mechanisms of disease pathogenesis and for preclinical testing.

Control and eradication of invasive mammals in Great Britain.

This paper provides a 'long view' of the eradication and control of invasive mammals by reviewing the management of 24 mammalian species that have been introduced into Great Britain since the Neolithic period and have subsequently established free-living populations in the wild. The approach provides examples of the issues faced when managing populations and examines some of the lessons that can be learned from successes and failures. The species are covered in the order of introduction, with the control/eradication of rabbit (Oryctolagus cuniculus), muskrat (Ondatra zibethicus), coypu (Myocastor coypus) and American mink (Mustela vison) considered in more detail. The species accounts are set within the context of commitments for the control of invasive alien species made by parties to the Convention on Biological Diversity and guidance provided by the International Union for Conservation of Nature and the Council of Europe. These have led to improvements in the process for assessing risks and co-ordinating action. However, despite some notable cases documented here, there have been few successful eradication programmes carried out in Europe. This paper argues that there is a case for building on the improved frameworks that are being developed in the United Kingdom and elsewhere and for being more ambitious with goals for the management of invasive alien species.

Prolactin regulation of testosterone secretion and testes growth in DLS rams at the onset of seasonal testicular recrudescence.

Our objective was to test the hypothesis that prolactin (PRL) acts at both the pituitary and testis levels to regulate testosterone secretion in the adult ram. The focus was on the mid-regression to mid-redevelopment stages of a photoperiod-condensed 'seasonal' testicular cycle. DLS rams (six per group) were given daily s.c. injections of bromocriptine (4 mg) or vehicle during the entire period. Serum PRL concentration in control rams peaked at 103.4+/-22.1 ng/ml in late regression and then steadily declined (P<0.01) to 19.5+/-4.3 ng/ml, whereas PRL in treated rams was always < or =4.0 ng/ml. Suppression of PRL tended (P<0.10) to increase the amplitude of natural LH pulses (transition stages) or reduce the number of LH receptors in the testis (regressed stage), although neither change disturbed testosterone levels in peripheral blood. These subtle changes were accompanied by significant (P<0.05) alterations in the capability of the pituitary to release LH (85% more) and of the testes to secrete testosterone (20% less). These effects of PRL were unmasked when rams were given highly stimulative i.v. injections of GNRH (single 3 microg dose) and NIH-oLH-S24 (three 5 microg doses given 20 min apart) respectively. PRL insufficiency also appeared to slow down the 'seasonal' rise in FSH secretion and slightly delayed (2 weeks) the times when the testes began to grow and were first significantly (P<0.05) enlarged from the regressed state. We conclude that PRL is an important part of the intricate regulation of the pituitary-gonadal system in moderately seasonal DLS rams.

PTEN signaling in brain: neuropathology and tumorigenesis.

Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a tumor suppressor that antagonizes the phosphatidylinositol-3-kinase (PI3K)/Akt/mTOR pathway by functioning as a lipid phosphatase. This ubiquitous and evolutionarily conserved signaling cascade influences numerous functions including cell growth, survival, proliferation, migration and metabolism. Inherited mutations in PTEN cause pleiotropic effects including cancer predisposition as well as a range of neurological abnormalities revealing specialized roles for PTEN in nervous system development and maintenance. Somatic mutations in PTEN occur frequently as late events in sporadic brain tumors. Mouse models based on Pten deletion in the brain have provided insights into the normal functions of Pten in the nervous system as well as the initiation and progression of gliomas. Compromised PTEN function may contribute to gliomagenesis through disrupted regulation of proliferation, migration, invasion, angiogenesis, stem cell self-renewal and regulation of other tumor suppressor pathways such as p53. Clinical findings in high-grade glioma suggest that PTEN gene alterations are associated with poor prognosis and may influence response to specific therapies. Emerging research using specific pharmacological inhibitors of the PI3K pathway may provide novel therapeutic options for the treatment of PTEN-deficient tumors.

Phosphatase and tensin homolog, deleted on chromosome 10 deficiency in brain causes defects in synaptic structure, transmission and plasticity, and myelination abnormalities.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway modulates growth, proliferation and cell survival in diverse tissue types and plays specialized roles in the nervous system including influences on neuronal polarity, dendritic branching and synaptic plasticity. The tumor-suppressor phosphatase with tensin homology (PTEN) is the central negative regulator of the PI3K pathway. Germline PTEN mutations result in cancer predisposition, macrocephaly and benign hamartomas in many tissues, including Lhermitte-Duclos disease, a cerebellar growth disorder. Neurological abnormalities including autism, seizures and ataxia have been observed in association with inherited PTEN mutation with variable penetrance. It remains unclear how loss of PTEN activity contributes to neurological dysfunction. To explore the effects of Pten deficiency on neuronal structure and function, we analyzed several ultra-structural features of Pten-deficient neurons in Pten conditional knockout mice. Using Golgi stain to visualize full neuronal morphology, we observed that increased size of nuclei and somata in Pten-deficient neurons was accompanied by enlarged caliber of neuronal projections and increased dendritic spine density. Electron microscopic evaluation revealed enlarged abnormal synaptic structures in the cerebral cortex and cerebellum. Severe myelination defects included thickening and unraveling of the myelin sheath surrounding hypertrophic axons in the corpus callosum. Defects in myelination of axons of normal caliber were observed in the cerebellum, suggesting intrinsic abnormalities in Pten-deficient oligodendrocytes. We did not observe these abnormalities in wild-type or conditional Pten heterozygous mice. Moreover, conditional deletion of Pten drastically weakened synaptic transmission and synaptic plasticity at excitatory synapses between CA3 and CA1 pyramidal neurons in the hippocampus. These data suggest that Pten is involved in mechanisms that control development of neuronal and synaptic structures and subsequently synaptic function.

Hematopoietic cytokine receptor signaling.

Hematopoiesis is the cumulative result of intricately regulated signaling pathways that are mediated by cytokines and their receptors. Proper culmination of these diverse pathways forms the basis for an orderly generation of different cell types. Recent studies conducted over the past 10-15 years have revealed that hematopoietic cytokine receptor signaling is largely mediated by a family of tyrosine kinases termed Janus kinases (JAKs) and their downstream transcription factors termed STATs (signal transducers and activators of transcription). Aberration in these pathways, such as that caused by the recently identified JAK2V617F mutation, is an underlying cause for diseases such as leukemias and other myeloproliferative disorders. This recent discovery, when coupled with the fact that STATs are activated by oncoproteins such as BCR-ABL, underscores the importance of the JAK-STAT pathway in both normal cellular development and disease states.

Blockade of mGluR1 receptor results in analgesia and disruption of motor and cognitive performances: effects of A-841720, a novel non-competitive mGluR1 receptor antagonist.

BACKGROUND AND PURPOSE: To further assess the clinical potential of the blockade of metabotropic glutamate receptors (mGluR1) for the treatment of pain. EXPERIMENTAL APPROACH: We characterized the effects of A-841720, a novel, potent and non-competitive mGluR1 antagonist in models of pain and of motor and cognitive function. KEY RESULTS: At recombinant human and native rat mGluR1 receptors, A-841720 inhibited agonist-induced calcium mobilization, with IC50 values of 10.7+/-3.9 and 1.0 +/- 0.2 nM, respectively, while showing selectivity over other mGluR receptors, in addition to other neurotransmitter receptors, ion channels, and transporters. Intraperitoneal injection of A-841720 potently reduced complete Freund's adjuvant-induced inflammatory pain (ED50 = 23 micromol kg(-1)) and monoiodoacetate-induced joint pain (ED50 = 43 micromol kg(-1)). A-841720 also decreased mechanical allodynia observed in both the sciatic nerve chronic constriction injury and L5-L6 spinal nerve ligation (SNL) models of neuropathic pain (ED50 = 28 and 27 micromol kg(-1), respectively). Electrophysiological studies demonstrated that systemic administration of A-841720 in SNL animals significantly reduced evoked firing in spinal wide dynamic range neurons. Significant motor side effects were observed at analgesic doses and A-841720 also impaired cognitive function in the Y-maze and the Water Maze tests. CONCLUSIONS AND IMPLICATIONS: The analgesic effects of a selective mGluR1 receptor antagonist are associated with motor and cognitive side effects. The lack of separation between efficacy and side effects in pre-clinical models indicates that mGluR1 antagonism may not provide an adequate therapeutic window for the development of such antagonists as novel analgesic agents in humans.

Ultrasound-guided foam sclerotherapy for the treatment of varicose veins.

BACKGROUND: The aim was to assess the early efficacy and complications of ultrasound-guided foam sclerotherapy (UGFS) in a cohort of patients with varicose veins. METHODS: Of 192 consecutive patients referred with varicose veins over 15 months, only 11 chose surgery; the rest underwent UGFS treatment. Polidocanol was foamed 1 : 3 with air. Under ultrasound control via butterfly or Seldinger cannulation, 1 per cent foam was injected into superficial veins and 3 per cent foam into saphenous trunks, up to a total volume of 14 ml. Outcome was defined as complete when occlusion of the saphenous trunk and/or over 85 per cent of the varicosities was achieved, and partial closure when less. RESULTS: In 163 legs, complete occlusion occurred after one intervention, a further 32 after a second, and one after a third (overall 91 per cent). Of the remainder, all other legs achieved partial occlusion after up to three interventions, apart from two legs with great saphenous vein (GSV) incompetence, which failed. All 23 legs with small saphenous veins had complete occlusion after one intervention compared with 64 of 97 legs with GSV incompetence (P < 0.010). Occlusion rates were also higher when the GSV was cannulated directly: 56 of 70 versus 8 of 27 (P < 0.001). CONCLUSIONS: UGFS achieved early complete occlusion safely in over 90 per cent of legs with varicose veins.

Use of an amorphous silicon electronic portal imaging device for multileaf collimator quality control and calibration.

Multileaf collimator (MLC) calibration and quality control is a time-consuming procedure typically involving the processing, scanning and analysis of films to measure leaf and collimator positions. Faster and more reliable calibration procedures are required for these tasks, especially with the introduction of intensity modulated radiotherapy which requires more frequent checking and finer positional leaf tolerances than previously. A routine quality control (QC) technique to measure MLC leaf bank gain and offset, as well as minor offsets (individual leaf position relative to a reference leaf), using an amorphous silicon electronic portal imaging device (EPID) has been developed. The technique also tests the calibration of the primary and back-up collimators. A detailed comparison between film and EPID measurements has been performed for six linear accelerators (linacs) equipped with MLC and amorphous silicon EPIDs. Measurements of field size from 4 to 24 cm with the EPID were systematically smaller than film measurements over all field sizes by 0.4 mm for leaves/back-up collimators and by 0.2 mm for conventional collimators. This effect is due to the gain calibration correction applied by the EPID, resulting in a 'flattening' of primary beam profiles. Linac dependent systematic differences of up to 0.5 mm in individual leaf/collimator positions were also found between EPID and film measurements due to the difference between the mechanical and radiation axes of rotation. When corrections for these systematic differences were applied, the residual random differences between EPID and film were 0.23 mm and 0.26 mm (1 standard deviation) for field size and individual leaf/back-up collimator position, respectively. Measured gains (over a distance of 220 mm) always agreed within 0.4 mm with a standard deviation of 0.17 mm. Minor offset measurements gave a mean agreement between EPID and film of 0.01+/-0.10 mm (1 standard deviation) after correction for the tilt of the EPID and small rotational misalignments between leaf banks and the back-up collimators used as a reference straight edge. Reproducibility of EPID measurements was found to be very high, with a standard deviation of <0.05 mm for field size and <0.1 mm for individual leaf/collimator positions for a 10x10 cm2 field. A standard set of QC images (three field sizes defined both by leaves only and collimators only) can be acquired in less than 20 min and analysed in 5 min.

Role of FSH, numbers of FSH receptors and testosterone in the regulation of inhibin secretion during the seasonal testicular cycle of adult rams.

The regulation of inhibin secretion has not been elucidated fully in male ruminants. The aim of this study was to determine the relative importance of FSH and testosterone concentrations, and FSH receptors, in the control of secretion of immunoactive inhibin in rams. In Expt 1, temporal changes in hormone concentrations and testicular FSH binding were determined for two groups of rams (n = 4) kept under opposite, alternating 4 month periods of long (16 h light:8 h dark) and short (8 h light:16 h dark) days. Testicular biopsies (1-2 g) were collected when the testes were regressed, redeveloping, redeveloped and regressing. In Expt 2, separate groups of rams (n = 4) kept under natural photoperiod (latitude 45 degrees 48 minutes N) were designated as controls or passively immunized (for 3 weeks) with sufficient oestradiol antiserum to increase testosterone secretion without altering LH and FSH; this was done when the testes were regressed (non-breeding season) and redeveloped (breeding season). In both groups of rams (Expt 1), 'seasonal' increases in FSH concentrations began a few weeks earlier than did increases in inhibin concentrations. FSH reached maximum concentrations during testicular recrudescence, whereas numbers of FSH receptors in the testis and circulatory inhibin concentrations did not reach peak values until the testes were fully developed. Numbers of FSH receptors per testis, but not FSH concentration, were positively correlated (r = 0.65) with inhibin concentrations across the four stages of the testicular cycle. Near the end of testicular recrudescence early in the breeding season (Expt 2), relatively high FSH concentration was associated with increased abundance of FSH receptor mRNA (90%) and number of receptors (45%) in the testis and increased inhibin concentrations (50%), compared with when the testes were regressed. Moderate, physiological increases in testosterone secretion in immunized rams did not affect inhibin in either season. These results indicate that: (i) FSH stimulation of immunoactive inhibin secretion by Sertoli cells as testes recrudesce is via increases in secretion (early) and cognate receptors (late); (ii) FSH upregulates the synthesis of its own receptor late in recrudescence; and (iii) the positive correlation (r = 0.70) observed between circulatory testosterone and immunoactive inhibin does not reflect a causal relationship.

Pten regulates neuronal soma size: a mouse model of Lhermitte-Duclos disease.

Somatic inactivation of PTEN occurs in different human tumors including glioblastoma, endometrial carcinoma and prostate carcinoma. Germline mutations in PTEN result in a range of phenotypic abnormalities that occur with variable penetrance, including neurological features such as macrocephaly, seizures, ataxia and Lhermitte-Duclos disease (also described as dysplastic gangliocytoma of the cerebellum). Homozygous deletion of Pten causes embryonic lethality in mice. To investigate function in the brain, we used Cre-loxP technology to selectively inactivate Pten in specific mouse neuronal populations. Loss of Pten resulted in progressive macrocephaly and seizures. Neurons lacking Pten expressed high levels of phosphorylated Akt and showed a progressive increase in soma size without evidence of abnormal proliferation. Cerebellar abnormalities closely resembled the histopathology of human Lhermitte-Duclos disease. These results indicate that Pten regulates neuronal size in vivo in a cell-autonomous manner and provide new insights into the etiology of Lhermitte-Duclos disease.

Combined effect of follicle-follicle interactions and declining follicle-stimulating hormone on murine follicle health in vitro.

Follicle selection occurs throughout an adult female's reproductive life, with selected, dominant follicle(s) developing to the preovulatory stage whereas the remaining, subordinate follicles within the growing cohort instead undergo atresia and die. To date, most research into follicle dominance has concentrated on its endocrine regulation, although it seems likely that intraovarian mechanisms are also involved in its regulation. We demonstrate here that the response of singly cultured murine follicles to declining concentrations of FSH depends on their developmental stage, with follicles at an earlier stage of development being much more susceptible than mature follicles to a lowering of FSH levels. We then extrapolate this information to follicle cocultures, in which a large dominant follicle was grown with a small subordinate follicle in a manner that maintained a dominant/subordinate relationship, with follicle health assessed by a terminal transferase-mediated 2'-deoxyuracil 5'-triphosphate nick end-labeled reaction on whole-follicle mounts. Our investigations show a combined negative effect of coculture and FSH withdrawal on small subordinate follicles, such that subordinate follicles cocultured with dominant follicles and subjected to a lowering of FSH levels during the culture period exhibit a greatly increased incidence of apoptosis in the granulosa cells (750% increase) compared with that exhibited by the dominant follicles (97% increase). We suggest that a similar interaction between endocrine and intraovarian factors regulates follicular dominance in vivo, such that dominant follicles, in addition to bringing about a fall in FSH levels via the hypothalamic-pituitary axis, exert local, direct effects on subordinate follicles, with both of these influences combining to induce atresia in subordinate follicles.

Nucleoside analogues as highly potent and selective inhibitors of herpes simplex virus thymidine kinase.

A series of carboxamide derivatives of 5'-amino-2',5'-dideoxy-5-ethyluridine has been prepared as inhibitors of HSV-TK (herpes simplex virus thymidine kinase). The most potent compounds were derived from xanthene, thioxanthene and dihydroanthracene carboxylic acids. The lead compounds show subnanomolar IC(50) values against HSV TKs.

PTEN induces G(1) cell cycle arrest and decreases cyclin D3 levels in endometrial carcinoma cells.

Inactivating mutations in the PTEN tumor suppressor gene occur in approximately 30-50% of endometrial carcinomas. PTEN is a phosphatase that negatively regulates the phosphoinositide 3-kinase signaling pathway, including the downstream effector AKT. To evaluate the role of PTEN in endometrial growth regulation, we expressed wild-type or mutant PTEN in endometrial carcinoma cell lines. As expected, expression of exogenous PTEN decreased levels of activated AKT in all cell lines examined. However, PTEN induced a G(1) cell cycle arrest specifically in endometrial carcinoma cells that lack endogenous wild-type PTEN. Growth of cells containing wild-type PTEN was unaffected by exogenous PTEN expression. Growth arrest required a functional phosphatase domain but not the PDZ interaction motif of PTEN. Overall levels of CIP/KIP and INK4 family members, the known inhibitory regulators of the G(1) phase of the cell cycle, were unchanged. However, PTEN induced a specific reduction of cyclin D3 levels and an associated increase in the amount of the inhibitor p27(KIP1) complexed with CDK2. Enforced expression of cyclin D3 abrogated the PTEN-induced cell cycle arrest. Although PTEN signaling directly regulates p27(KIP1) levels in some settings, in endometrial carcinoma cells, PTEN expression indirectly regulated p27(KIP1) activity by modulating levels of cyclin D3. These data support multiple mechanisms of PTEN-induced cell cycle arrest.

Characterization of an alternatively spliced AATYK mRNA: expression pattern of AATYK in the brain and neuronal cells.

The AATYK gene encodes a tyrosine kinase whose expression is up-regulated during the apoptosis and differentiation of 32Dcl3 myeloblastic cells. Because high levels of AATYK mRNA have also been detected in the brain, and because these transcripts differ in size from that observed in the 32Dcl3 cell line, it was of interest to determine whether this gene encodes mRNAs that are alternatively spliced and whether these mRNAs are expressed in a tissue-specific manner. We have isolated a novel, alternatively spliced AATYK mRNA using cDNA library screening and RT-PCR, whose expression is readily detected in the brain but not myeloid cells. Western blot analysis revealed that the AATYK protein was expressed in virtually all regions of the adult rat brain in which neurons are present, including olfactory bulb, forebrain, cortex, midbrain, cerebellum and pons. Immunohistochemical labeling of adult brain sections showed the highest levels of AATYK expression in the cerebellum and olfactory bulb. Expression of AATYK was also up-regulated as a function of RA-induced neuronal differentiation of p19 embryonal carcinoma cells, supporting a role for this protein in mature neurons and neuronal differentiation.

Parathyroid hormone, haematological and biochemical parameters in relation to dental disease and husbandry in rabbits.

During a two-year period between 1995 and 1997, over 80 blood samples were collected from pet rabbits in order to investigate an apparent osteodystrophy affecting the skulls of rabbits with acquired dental disease. A series of haematological and biochemical analyses relating to calcium metabolism were performed and samples were taken for parathyroid hormone (PTH) assay. The rabbits were categorised according to the condition of their teeth and the manner in which the pets were kept. PTH concentrations were higher and calcium concentrations lower in hutch-kept rabbits with advanced dental disease in comparison with those kept in free-range conditions. No dental problems were detected in the free-range rabbits on radiological or clinical examination. During the course of the study, differences in haematological pictures and albumin values emerged among rabbits kept under the different husbandry regimes. Complete blood counts from free-range rabbits were comparable with laboratory reference ranges, whereas there were significantly lower red cell and lymphocyte counts in rabbits exhibiting advanced dental disease. Serum albumin values were significantly higher in rabbits kept in free-range conditions than in those with advanced dental disease or those unaffected by dental disease but kept in hutches. Rabbits kept in hutches showed trends towards anaemia and lymphopenia. Results indicated that acquired dental disease of pet rabbits is related to husbandry and is associated with alterations in calcium metabolism.