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John Mullins

Publications and source records attributed to John Mullins.

6 recordsLinked to original sources
PubMed2005

Comparative proteomic analysis using samples obtained with laser microdissection and saturation dye labelling.

Comparative proteomic methods are rapidly being applied to many different biological systems including complex tissues. One pitfall of these methods is that in some cases, such as oncology and neuroscience, tissue complexity requires isolation of specific cell types and sample is limited. Laser microdissection (LMD) is commonly used for obtaining such samples for proteomic studies. We have combined LMD with sensitive thiol-reactive saturation dye labelling of protein samples and 2-D DIGE to identify protein changes in a test system, the isolated CA1 pyramidal neurone layer of a transgenic (Tg) rat carrying a human amyloid precursor protein transgene. Saturation dye labelling proved to be extremely sensitive with a spot map of over 5,000 proteins being readily produced from 5 mug total protein, with over 100 proteins being significantly altered at p < 0.0005. Of the proteins identified, all showed coherent changes associated with transgene expression. It was, however, difficult to identify significantly different proteins using PMF and MALDI-TOF on gels containing less than 500 mug total protein. The use of saturation dye labelling of limiting samples will therefore require the use of highly sensitive MS techniques to identify the significantly altered proteins isolated using methods such as LMD.

Alzheimer Disease↗
PubMed2005

On the verification of intransitive noninterference in mulitlevel security.

We propose an algorithmic approach to the problem of verification of the property of intransitive noninterference (INI), using tools and concepts of discrete event systems (DES). INI can be used to characterize and solve several important security problems in multilevel security systems. In a previous work, we have established the notion of iP-observability, which precisely captures the property of INI. We have also developed an algorithm for checking iP-observability by indirectly checking P-observability for systems with at most three security levels. In this paper, we generalize the results for systems with any finite number of security levels by developing a direct method for checking iP-observability, based on an insightful observation that the iP function is a left congruence in terms of relations on formal languages. To demonstrate the applicability of our approach, we propose a formal method to detect denial of service vulnerabilities in security protocols based on INI. This method is illustrated using the TCP/IP protocol. The work extends the theory of supervisory control of DES to a new application domain.

Algorithms↗
PubMed2003

Murine Denys-Drash syndrome: evidence of podocyte de-differentiation and systemic mediation of glomerulosclerosis.

Denys-Drash syndrome (DDS) is caused by dominant mutations of the Wilms' tumour suppressor gene, WT1, and characterized by a nephropathy involving diffuse mesangial sclerosis, male pseudohermaphroditism and/or Wilms' tumourigenesis. Previously, we reported that heterozygosity for the Wt1tmT396 mutation induces DDS in heterozygous and chimeric (Wt1tmT396/+<-->+/+) mice. In the present study, the fate of Wt1 mutant cells in chimeric kidneys was assessed by in situ marker analysis, and immunocytochemistry was used to re-examine the claim that glomerulosclerosis (GS) is caused by loss of WT1 and persistent Pax-2 expression by podocytes. Wt1 mutant cells colonized glomeruli efficiently, including podocytes, but some sclerotic glomeruli contained no detectable Wt1 mutant cells. The development of GS was preceded by widespread loss of ZO-1 signal in podocytes (even in kidneys where <5% of glomeruli contained Wt1 mutant podocytes), increased intra-renal renin expression, and de novo podocyte TGF-beta1 expression, but not podocyte Pax-2 expression or loss of WT1, synaptopodin, alpha-actinin-4 or nephrin expression. However, podocytes in partially sclerotic glomeruli that still expressed WT1 at high levels showed reduced vimentin expression, cell cycle re-entry, and re-expressed desmin, cytokeratin and Pax-2. The results suggest that: (i) GS is not due to loss of WT1 expression by podocytes; (ii) podocyte Pax-2 expression reflects re-expression rather than persistent expression, and is the consequence of GS; (iii) GS is mediated systemically and the mechanism involves activation of the renin-angiotensin system; and (iv) podocytes undergo typical maturational changes but subsequently de-differentiate and revert to an immature phenotype during disease progression.

Animals↗
PubMed2003

Enhanced long-term potentiation in the hippocampus of rats expressing mutant presenillin-1 is age related.

Electrophysiological recordings were made from Fischer rats engineered to express the human presenilin 1 gene carrying the M146V mutation. Extracellular recordings of field excitatory post-synaptic potential (EPSPs) were made to investigate EPSP properties, paired pulse responses, posttetanic potentiation, and long-term potentiation in the stratum radiatum and dentate gyrus of hippocampal slices maintained in vitro. Transgenic rats aged approximately 6 months showed no differences from their wild-type littermates in any of these properties. However, at 18 months, long-term potentiation in the CA1 was facilitated in the transgenic rats with a different pattern of synaptic enhancement. No changes were observed in paired pulse facilitation (PPF) or post-tetanic potentiation (PPT) and no changes were seen in the dentate gyrus. Field potential amplitudes were significantly greater and PPF was enhanced in the CA1 of all older rats. Intracellular recordings from CA1 pyramidal cells of the older group of rats revealed no differences in the passive or active membrane properties of cells in the two groups, but intracellularly recorded EPSPs were significantly longer.

Aging↗
PubMed2002

Cord-like mosaic patches in the adrenal cortex are fractal: implications for growth and development.

Organogenesis proceeds rapidly and faithfully during fetal development. The process includes generation of parenchyma, followed by organization into functional tissues. The method by which the growth of organ parenchyma is regulated is not known, but insight into this regulation has been obtained by studying mosaic tissues of experimental chimeras and transgenic mosaics. The patterns revealed by this procedure offer an indication of how the parenchyma was generated. In the liver, the pattern appears as islands of one cell type in a sea of the other cell type, while in the adrenal cortex the pattern is one of alternating cords of one cell type adjacent to the other cell type. We have established previously that mosaic patches in the liver are fractal. The fractal dimensions of patches in the liver are consistent with an iterative, recursive growth model with simple stereotypical division rules. Here we report that the patches in mosaic adrenal cortex of the mouse and rat are also fractal and that the fractal dimension of the surface of the patches is lower than that in the liver. Fetal development and fractal dimensions of adrenal cortical mosaic patches are consistent with an algorithmic cell division model in which parenchymal growth is constrained to edges of growth centers forcing cord structures to form. Fractal analysis of the geometry of mosaic patches in tissues of experimental chimeras is helpful in constructing hypotheses of organ growth.

Adrenal Cortex↗