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Biomedical subjects

R McKay

Publications and source records attributed to R McKay.

At least 19 recordsLinked to original sources
PubMed2000

Stem cells and the cellular organization of the brain.

The modern field of neuroscience emerged in the hands of neuroanatomists at the end of the last century. They recognized that information flows through arrays of cells and that the structure of the cells shapes the passage of information through the brain. In the middle of this century, the role of ion flux in the movement of information along axons was established. The electron microscope and the microelectrode helped to explain information transfer at the synapse, and study of the modulation of synaptic strength is currently a major area of neuroscience research. The origin of the many types of neuron seen by the early anatomists remains enigmatic, but the solution to this mystery is now emerging. The identification of stem cells and of the mechanisms that control their differentiation into distinct neuron types will contribute to a new understanding of both the origin of neuronal types and neuronal circuits. The current enthusiasm for stem cells is stimulated by interest from both the academic and nonacademic communities. These enthusiasts recognize a simple truth, that the cells are smart; they are the agents that control cell number and type in the brain. They create the structure that generates the higher order brain function. If these premises are true, then in the new century our understanding of the nervous system will be transformed by the facts of stem cell biology. J. Neurosci. Res. 59:298-300, 2000. Published 2000 Wiley-Liss, Inc.

Animals↗
PubMed2000

Antisense methods for discrimination of phenotypic properties of closely related gene products: Jun kinase family.

Methods for the selection and characterization of antisense oligonucleotides for specifically eliminating closely related gene family members are available. High-throughput semiautomated methods using 96-well plate formats and array technology and improved assays are under active development that will streamline many steps and will likely merge. Second-generation 20-mer antisense phosphorothioate oligonucleotides containing 2'-methoxyethyl groups at the first and last 6 nucleotides with improved nuclease resistance and RNA affinity are becoming available.

Animals↗
PubMed1999

Management of diabetic retinopathy by Australian optometrists. Working Group on Evaluation of NHMRC Retinopathy Guideline Distribution. National Health and Medical Research Council.

PURPOSE: To describe current practices related to the management of diabetic retinopathy by Australian optometrists. SETTING AND METHODS: A two-page self-administered questionnaire was mailed to a random sample of 504 Australian optometrists. The survey included questions about the practice (such as size and location); current practice with regard to management of patients with diabetic retinopathy; barriers to use of dilating drops; and a number of patient scenarios related to screening, follow-up and treatment of diabetic retinopathy. RESULTS: Completed questionnaires were returned by 407 of the 473 eligible optometrists (86%). They had been practising optometry between 1 and 50 years (median 14). Of the 243 optometrists who provided details about the location of their practices, 145 (37%) had at least one of their practices in a rural area. The estimated percentage of patients with diagnosed diabetes ranged from 0.5 to 40% (median = 5.0%). Three-hundred and twenty-two optometrists (79%) reported that they would often or almost always ask new patients over the age of 40 whether they have diabetes. The majority of optometrists (n = 387, 95%) would often or almost always ask their new patients with diabetes about their control of blood glucose levels and the majority of optometrists (n = 330, 81%) would often or almost always tell their patients with diabetes about the importance of strict glucose control in delaying retinopathy. The most common barrier to dilated ophthalmoscopy was patients not wanting to be dilated, with 38.1% of optometrists reporting this to be a moderate or major barrier. The next most common barrier was fear of precipitating angle closure glaucoma; 17.1% of optometrists reported this to be a moderate or major barrier CONCLUSION: The National Health and Medical Research Council (NHMRC) guidelines for the management of diabetic retinopathy are timely in relation to the expressed desire of Australian optometrists to learn more about management of diabetic retinopathy These data will be used prospectively to assess changes in management of patients with diabetic retinopathy as a result of the release of the NHMRC guidelines.

Australia↗
PubMed1999

The Jun kinase 2 isoform is preferentially required for epidermal growth factor-induced transformation of human A549 lung carcinoma cells.

We have previously found that epidermal growth factor (EGF) mediates growth through the Jun N-terminal kinase/stress-activated kinase (JNK/SAPK) pathway in A549 human lung carcinoma cells. As observed here, EGF treatment also greatly enhances the tumorigenicity of A549 cells, suggesting an important role for JNK in cancer cell growth (F. Bost, R. McKay, N. Dean, and D. Mercola, J. Biol. Chem. 272:33422-33429, 1997). Several isoforms families of JNK, JNK1, JNK2, and JNK3, have been isolated; they arise from alternative splicing of three different genes and have distinct substrate binding properties. Here we have used specific phosphorothioate oligonucleotides targeted against the two major isoforms, JNK1 and JNK2, to discriminate their roles in EGF-induced transformation. Multiple antisense sequences have been screened, and two high-affinity and specific candidates have been identified. Antisense JNK1 eliminated steady-state mRNA and JNK1 protein expression with a 50% effective concentration (EC50) of <0.1 microM but did not alter JNK2 mRNA or protein levels. Conversely, antisense JNK2 specifically eliminated JNK2 steady-state mRNA and protein expression with an EC50 of 0.1 microM. Antisense JNK1 and antisense JNK2 inhibited by 40 and 70%, respectively, EGF-induced total JNK activity, whereas sense and scrambled-sequence control oligonucleotides had no effect. The elimination of mRNA, protein, and JNK activities lasted 48 and 72 h following a single Lipofectin treatment with antisense JNK1 and JNK2, respectively, indicating sufficient duration for examining the impact of specific elimination on the phenotype. Direct proliferation assays demonstrated that antisense JNK2 inhibited EGF-induced doubling of growth as well as the combination of active antisense oligonucleotides did. EGF treatment also induced colony formation in soft agar. This effect was completely inhibited by antisense JNK2 and combined-antisense treatment but not altered by antisense JNK1 alone. These results show that EGF doubles the proliferation (growth in soft agar as well as tumorigenicity in athymic mice) of A549 lung carcinoma cells and that the JNK2 isoform but not JNK1 is utilized for mediating the effects of EGF. This study represents the first demonstration of a cellular phenotype regulated by a JNK isoform family, JNK2.

Agar↗
PubMed1998

Resolution of the heterogeneous fluorescence in multi-tryptophan proteins: ascorbate oxidase.

Ascorbate oxidase is a copper-containing enzyme which catalyzes a redox reaction between vitamin C and molecular oxygen. The protein, which shows a complex tertiary structure, is an homodimer of monomers, each containing three domains and 14 tryptophan residues. Recently, we have demonstrated by spectroscopic and ultracentrifugation techniques the existence of a stable dimeric intermediate along the unfolding pathway of this enzyme [Mei, G., Di Venere, A., Buganza, M., Vecchini, P., Rosato, N. & Finazzi Agrò, A. (1997) Biochemistry 36, 10917-10922]. In this study, the steady-state and dynamic fluorescence features of ascorbate oxidase have been exploited in order to find a way of monitoring the individual subsystems of the protein. The fluorescence intensity and anisotropy upon excitation at 295 nm are extremely sensitive functions of the emission wavelength, indicating a great heterogeneity of the system. The emission decay collected through a cut-off filter can be analyzed in terms of two continuous distributions of lifetimes. Using a monochromator in emission or an optical multichannel analyzer, the two distributions may be attributed to distinct components of the fluorescence spectrum. Differential quenching by cesium chloride also confirmed that the several tryptophan residues present in the protein structure may be grouped into two main classes, each with a different environment. Once the complex fluorescence decay of ascorbate oxidase was analyzed and resolved, a comparison with the crystallographic data allowed a first, approximate attribution of the protein spectroscopic properties to some of the tryptophan residues. This might provide a powerful tool of investigation about the role of definite segments of the protein in its three-dimensional structure and catalytic activity. Furthermore, the methodology set up for ascorbate oxidase can be usefully extended to other multitryptophan proteins.

Ascorbate Oxidase↗
PubMed1998

Open-heart correction of tetralogy of Fallot in an acyanotic dog.

Tetralogy of Fallot was diagnosed in an acyanotic 11-month-old dog. Predicted pressure gradient across the pulmonic valve, as assessed by use of continuous wave Doppler echocardiography, was 94.5 mm Hg. Bidirectional shunting was identified by means of selective angiography. Open-heart correction was performed, using a transatrial approach with limited ventriculotomy and cardiopulmonary bypass. The hypertrophied infundibulum was resected, the ventricular septal defect was closed primarily, and a transannular pericardial patch graft was applied. Pressure gradients across the pulmonic valve were 52.9 and 22.8 mm Hg 2 weeks and 4 months after surgery, respectively. Advances in cardiopulmonary bypass, anesthetic management, and use of the transatrial approach may improve the success of open-heart correction of tetralogy of Fallot in dogs.

Animals↗
PubMed1998

Report of two sibs with Knobloch syndrome (encephalocoele and viteroretinal degeneration) and other anomalies.

We report on two sibs with high myopia, vitreoretinal degeneration (VRD), and occipital encephalocoele or scalp lesion. We review the literature on Knobloch syndrome, discuss possible causes, and suggest a possible involvement of mesoderm in the morphogenesis. One case presents with very early onset of severe eye disease, whereas the other is notable for the very mild scalp defect. In addition, both appear to have an unusual pulmonary lymphatic condition.

Abnormalities, Multiple↗
PubMed1998

Fibromuscular Dysplasia of the Aorta Presenting as Multiple Recurrent Thoracic Aneurysms

A 17-year-old boy, who had undergone resection of aortic coarctation with a large saccular aneurysm 10 years previously, developed recurrent aneurysms above and below the site of a Gore-Tex interposition graft. These were resected using femoral-femoral bypass, and the upper thoracic aorta was replaced with a Hemashield Dacron tube. Histology of the aorta showed fibromuscular dysplasia. In addition to aortic dissection and aortic rupture, such patients may be at risk of forming further aneurysms.

Journal Article↗
PubMed1998

Plasticity and stem cells in the vertebrate nervous system.

How and when do vertebrate neural precursor cells choose their fates? While some studies suggest a series of commitments on the road to fate choice, many recent experiments indicate that precursor fate choices can often be changed. Additionally, the identification of common gene control mechanisms in precursors suggest that these cells share fundamental properties throughout development.

Animals↗
PubMed1998

Stem cells and neurogenesis in the adult brain.

A new interest in replacing neurons lost to trauma or disease has been generated by findings that challenge the traditional view of the static (irreparable) adult brain. The discovery of stem cells and neurogenesis in the adult central nervous system is responsible for much of this interest.

Adult↗
PubMed1997

The JUN kinase/stress-activated protein kinase pathway is required for epidermal growth factor stimulation of growth of human A549 lung carcinoma cells.

Epidermal growth factor (EGF) plays a major role in non-small cell lung cancer cell autocrine growth and has been reported to activate the JUN kinase/stress-activated protein kinase (JNK/SAPK) pathway in model cells. Activation of JNK/SAPK leads to the phosphorylation of c-JUN protooncogene on serines 63 and 73. This mechanism is required for and cooperates in the transformation of rat embryo fibroblasts by Ha-RAS. However, the function of JNK/SAPK in human tumor growth is unknown. We have tested several lung carcinoma cell lines. All exhibited UV-C-inducible JNK/SAPK activity; two exhibited constitutive activity in low serum, and two (M103 and A549) exhibited EGF-inducible JNK/SAPK activity. In A549 cells, EGF induced a rapid and prolonged (up to 24 h) activation of the JNK/SAPK pathway that correlated with a 150-190% growth stimulation. Stably transfected clones of A549 cells expressing c-JUN(S63A,S73A), a transdominant inhibitor of c-JUN, completely blocked the EGF-stimulated proliferation effect but did not alter the basal proliferation rate. Consistent with these results JNK antisense oligonucleotides targeted to JNK1 and JNK2 entirely eliminated the EGF-stimulated JNK/SAPK activity and blocked EGF-stimulated growth but not basal growth. In contrast, specific inhibition of the RAF/ERK pathway by PD98059 (MEK1 inhibitor) completely blocked ERK activation by EGF and basal cell growth but not EGF-stimulated growth, thereby dissociating the growth-promoting roles of each pathway. Our observations indicate, for the first time, that JNK/SAPK may be a preferential effector pathway for the growth properties of EGF in A549 cells.

Animals↗
PubMed1997

Stem cells in the central nervous system.

In the vertebrate central nervous system, multipotential cells have been identified in vitro and in vivo. Defined mitogens cause the proliferation of multipotential cells in vitro, the magnitude of which is sufficient to account for the number of cells in the brain. Factors that control the differentiation of fetal stem cells to neurons and glia have been defined in vitro, and multipotential cells with similar signaling logic can be cultured from the adult central nervous system. Transplanting cells to new sites emphasizes that neuroepithelial cells have the potential to integrate into many brain regions. These results focus attention on how information in external stimuli is translated into the number and types of differentiated cells in the brain. The development of therapies for the reconstruction of the diseased or injured brain will be guided by our understanding of the origin and stability of cell type in the central nervous system.

Animals↗
PubMed1997

Definition by specific antisense oligonucleotides of a role for protein kinase C alpha in expression of differentiation markers in normal and neoplastic mouse epidermal keratinocytes.

Epidermal keratinocyte differentiation is a tightly regulated, stepwise process that requires protein kinase C (PKC) activation. Studies using cultured mouse keratinocytes induced to differentiate with Ca2+ have indirectly implicated the alpha isoform of PKC in upregulation of "late" (granular cell) epidermal differentiation markers. Activation of this isoform is also implicated in the suppression of "early" differentiation markers keratin (K) 1 and 10 that characterizes the neoplastic phenotype produced by the v-Ha-ras oncogene. We used antisense oligonucleotides (AS) to directly address the role of PKC alpha in regulating expression of these markers in normal and v-Ha-ras-transduced primary keratinocytes and a keratinocyte cell line (SP-1) containing an activating mutation of the c-Ha-ras gene. Transfection of PKC alpha AS reduced the PKC alpha protein level in a dose-dependent manner, with a maximum effect at doses of 100 nM or higher. Immunoblot analysis with antibodies against PKC alpha, PKC delta, PKC epsilon, and PKC eta confirmed that PKC alpha AS selectively reduced the level of PKC alpha but not the other isoforms. In vitro kinase assays also revealed suppression of Ca(2+)-dependent PKC activity, which is the PKC alpha activity in this cell type, after transfection of PKC alpha AS. When PKC alpha AS-treated normal keratinocytes were stimulated to terminally differentiate with Ca2+, induction of the late differentiation markers loricrin, filaggrin, and SPR-1, as well as transglutaminase K mRNA, was suppressed when compared with their induction in scrambled AS-treated controls. In neoplastic v-Ha-ras-transduced keratinocytes and SP-1 cells, transfection of PKC alpha AS, but not the scrambled AS control, selectively downregulated PKC alpha and restored differentiation-specific expression of K1. These findings directly confirm that PKC alpha is an important component of the signaling pathway regulating terminal differentiation of normal keratinocytes and that activation of PKC alpha contributes to the altered differentiation program of neoplastic murine keratinocytes.

Animals↗