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Biomedical subjects

P J Mitchell

Publications and source records attributed to P J Mitchell.

At least 37 records · Page 2Linked to original sources
PubMed2001

The angiosome concept applied to arteriovenous malformations of the head and neck.

Arteriovenous malformations remain a difficult clinical problem. There is very little understanding of the underlying pathogenesis of these lesions, and therapy frequently involves considerable risks with suboptimal outcomes. Recently, a comprehensive description of the angiosomes of the head and neck was completed in the authors' unit. It was noticed that the location of several clinically observed arteriovenous malformations in the head and neck seemed to correspond to the anatomic location of the choke anastomotic zones linking the angiosomes. Therefore, selective clinical angiograms were compared with those from the authors' previously performed fresh cadaver injection studies, in which they defined the angiosomes of the head and neck. In each patient, the location of the arteriovenous malformation corresponded directly to the choke vessel anastomotic zone linking two or more adjacent angiosomes. Clinical and pathologic ramifications of this observation are discussed.

Adolescent↗
PubMed2001

The AP-2 transcription factor is required for joint formation and cell survival in Drosophila leg development.

Flies mutant for the Drosophila homologue of the mammalian transcription factor AP-2 show a severe reduction in leg length and fail to develop joint structures. Presumptive joint cells express dAP-2 in response to Notch signaling. dAP-2 is required for joint cell differentiation and can induce formation of supernumerary joints when misexpressed. Although dAP-2 is expressed only in presumptive joint cells, its activity is required to support cell survival in the entire leg segment. Taken together, our data indicate that dAP-2 is an important mediator of Notch activity in leg development.

Animals↗
PubMed2001

Drosophila transcription factor AP-2 in proboscis, leg and brain central complex development.

We report loss- and gain-of-function analyses that identify essential roles in development for Drosophila transcription factor AP-2. A mutagenesis screen yielded 16 lethal point mutant alleles of dAP-2. Null mutants die as adults or late pupae with a reduced proboscis, severely shortened legs (approximately 30% of normal length) lacking tarsal joints, and disruptions in the protocerebral central complex, a brain region critical for locomotion. Seven hypomorphic alleles constitute a phenotypic series yielding hemizygous adults with legs ranging from 40-95% of normal length. Hypomorphic alleles show additive effects with respect to leg length and viability; and several heteroallelic lines were established. Heteroallelic adults have moderately penetrant defects that include necrotic leg joints and ectopic growths (sometimes supernumerary antennae) invading medial eye territory. Several dAP-2 alleles with DNA binding domain missense mutations are null in hemizygotes but have dominant negative effects when paired with hypomorphic alleles. In wild-type leg primordia, dAP-2 is restricted to presumptive joints. Ectopic dAP-2 in leg discs can inhibit but not enhance leg elongation indicating that functions of dAP-2 in leg outgrowth are region restricted. In wing discs, ectopic dAP-2 cell autonomously transforms presumptive wing vein epithelium to ectopic sensory bristles, consistent with an instructive role in sensory organ development. These findings reveal multiple functions for dAP-2 during morphogenesis of feeding and locomotor appendages and their neural circuitry, and provide a new paradigm for understanding AP-2 family transcription factors.

Alleles↗
PubMed2001

Endovascular management of vein of Galen aneurysmal malformations presenting in the neonatal period.

BACKGROUND AND PURPOSE: Neonates with vein of Galen aneurysmal malformations (VGAMs) presenting with cardiac failure have high morbidity and mortality, and outcomes are significantly better in those presenting in later childhood. Neurologic outcomes in survivors are perceived to be uniformly poor, which may lead to the neonate being denied treatment. We assessed outcomes of modern neonatal intensive care and endovascular embolization in a consecutive series of such neonates presenting with cardiac failure. METHODS: Between 1996 and 1998, five infants (three male, two female) were diagnosed with symptomatic VGAMs in the first week of life, four of whom had intractable, high-output cardiac failure and underwent initial endovascular treatment. There were 15 endovascular procedures and one neurosurgical clipping in these five patients. Transarterial and transvenous routes were required, using multiple embolic agents. We emphasized the use of sonographically guided, percutaneous transtorcular-venous-access, moveable-core guidewire as an embolic agent; routine MR imaging; and MR angiography. RESULTS: Immediate outcomes included control of cardiac failure with normal neurologic function in four (80%) patients and one (20%) death from intractable cardiac failure. On follow-up examination, three (60%) infants showed no evidence of neurologic abnormality or cardiac failure; one (20%) infant showed moderate developmental delay. Two have had no further shunting on angiography, one has minimal flow, and one is awaiting follow-up imaging. CONCLUSION: Endovascular therapy with modern neuroanesthetic and neurointensive care can provide good outcomes even in the highest-risk neonates with VGAMs and cardiac failure. If medical management of cardiac failure fails, and there is no evidence of gross cerebral parenchymal damage on imaging, urgent endovascular treatment is feasible and can reduce the almost-100% mortality otherwise expected, without invariably severe morbidity. Use of multiple embolization strategies in multiple stages usually is necessary in these patients, and novel approaches and embolic agents may be necessary.

Cerebral Veins↗
PubMed2000

A novel adaptor-like protein which is a substrate for the non-receptor tyrosine kinase, BRK.

The brk gene encodes a non-receptor tyrosine kinase that has been found to be overexpressed in approximately two thirds of breast tumours. Using a yeast two-hybrid based screen, we have cloned cDNAs encoding a novel protein, BKS, that is a substrate for the kinase activity of BRK and has the characteristics of an adaptor protein. BKS possesses an N-terminal PH-like domain followed by an SH2-like domain. In co-transfection experiments, high levels of phosphotyrosine were observed on BKS and BRK was found to be associated with BKS, both of which were dependent on the catalytic activity of BRK. The phosphorylation of and association with BKS by BRK was also dependent on the SH2-like domain present within BKS. In addition, BKS recruited an unidentified 100 kDa protein that was also phosphorylated on tyrosine residues in the presence of BRK. We have determined that the BKS protein is expressed in most adult human tissues. Oncogene (2000) 19, 4273 - 4282

Adaptor Proteins, Signal Transducing↗
PubMed2000

FGF5 stimulates expansion of connective tissue fibroblasts and inhibits skeletal muscle development in the limb.

FGF5 is expressed in the mesenchyme and skeletal muscle of developing and adult mouse limbs. However, the function of FGF5 during development of the limb and limb musculature is unknown. To elucidate the inherent participation of FGF5 during limb organogenesis, a retroviral delivery system (RCAS) was used to overexpress human FGF5 throughout developing hind limb of chicken embryos. Misexpression of the soluble growth factor severely inhibited the formation of mature myocytes. Limbs infected with RCAS-FGF5 contained smaller presumptive muscle masses as evidenced by a decrease in MyoD and myosin heavy chain expressing cells. In contrast, ectopic expression of FGF5 significantly stimulated proliferation and expansion of the tenascin-expressing, connective-tissue fibroblast lineage throughout the developing limb. Histological analysis demonstrated that the increase in tenascin immunostaining surrounding the femur, ileum, and pubis in the FGF5 infected limbs corresponded to the fibroblasts forming the stacked-cell perichondrium. Furthermore, pulse labeling experiments with the thymidine analog, BrdU, revealed that the increased size of the perichondrium was attributable to enhanced cell proliferation. These results support a model whereby FGF5 acts as a mitogen to stimulate the proliferation of mesenchymal fibroblasts that contribute to the formation of connective tissues such as the perichondrium, and inhibits the development of differentiated skeletal muscle. These results also contend that FGF5 is a candidate mediator of the exclusive spatial patterning of the hind limb connective tissue and skeletal muscle.

Animals↗
PubMed2000

Ectopic expression of luteinizing hormone-releasing hormone and peripherin in the respiratory epithelium of mice lacking transcription factor AP-2alpha.

The vertebrate transcription factor activator protein-2 (AP-2alpha) is involved in craniofacial morphogenesis. In the nasal placode AP-2alpha expression delineates presumptive respiratory epithelia from olfactory epithelia, with AP-2alpha expression restricted to the anterior region of the respiratory epithelium (absent from the olfactory epithelium) at later stages. To address the role AP-2alpha plays in differentiation of cell groups in the nasal placode, the spatiotemporal expression pattern of four markers normally associated with olfactory epithelial structures was analyzed in mice lacking AP-2alpha. These markers were the intermediate filament protein peripherin, the neuropeptide luteinizing hormone-releasing hormone (LHRH), the neural cell adhesion molecule (NCAM) and the olfactory transcription factor Olf-1. Development of cells expressing these markers was similar in both genotypes until embryonic day 12.5 (E12.5), indicating that the main olfactory epithelium and olfactory pit formation was normal. At E13.5 in mutant mice, ectopic LHRH neurons and peripherin axons were detected in respiratory epithelial areas, areas devoid of Olf-1 and NCAM staining. Over the next few days, an increase in total nasal LHRH neurons occurred. The increase in nasal LHRH neurons could be accounted for by LHRH neurons arising and migrating out of respiratory epithelial regions on peripherin-positive fibers. These results indicate that AP-2alpha is not essential for the separation of the olfactory and respiratory epithelium from the nasal placode and is consistent with AP-2alpha preventing recapitulation of developmental programs within the respiratory epithelium that lead to expression of LHRH and peripherin phenotypes.

Animals↗
PubMed2000

Failure in post-transcriptional processing is a possible inactivation mechanism of AP-2alpha in cutaneous melanoma.

The loss of transcription factor AP-2alpha expression has been shown to associate with tumourigenicity of melanoma cell lines and poor prognosis in primary cutaneous melanoma. Altogether these findings suggest that the gene encoding AP-2alpha (TFAP2A) acts as a tumour suppressor in melanoma. To learn more of AP-2alpha's down-regulation mechanisms, we compared the immunohistochemical AP-2alpha protein expression patterns with the corresponding mRNA expression detected by in situ hybridization in 52 primary melanomas. Of the 25 samples with AP-2alpha protein negative areas, 16 (64%) expressed mRNA throughout the consecutive section. Nine specimens (36%) contained equally mRNA- and protein-negative areas, suggesting that the loss of AP-2alpha protein associated with lack of the mRNA transcript. The highly AP-2alpha protein-positive tumours (n = 27) were concordantly mRNA positive in 25 (92.6%) cases. Thirteen primary tumours were further analysed using microsatellite markers D6S470 and D6S263 for loss of heterozygosity (LOH) of a locus harbouring TFAP2A. LOHs or chromosome 6 monosomy were found in four out of five (80%) informative AP-2alpha mRNA- and protein-negative tumour areas, but also within five out of 13 (38%) informative AP-2alpha mRNA-positive tumour areas. This chromosome region is thus suggestive of harbouring a putative tumour suppressor gene of cutaneous melanoma, but this referring specifically to TFAP2A could not be completely verified in this analysis. We conclude that a failure in post-transcriptional processing of AP-2alpha is a possible inactivation mechanism of AP-2alpha in cutaneous melanoma.

DNA-Binding Proteins↗
PubMed2000

Expression of transcription factor AP-2alpha predicts survival in epithelial ovarian cancer.

The 52-kDa activator protein (AP)-2 is a DNA-binding transcription factor which has been reported to have growth inhibitory effects in cancer cell lines and in human tumours. In this study the expression of AP-2alpha was analysed in 303 epithelial ovarian carcinomas by immunohistochemistry (IHC) with a polyclonal AP-2alpha antibody and its mRNA status was determined by in situ hybridization (ISH) and reverse transcriptase-polymerase chain reaction (RT-PCR). The immunohistochemical expression of AP-2alpha was correlated with clinicopathological variables, p21/WAF1 protein expression and survival. In normal ovaries, epithelial cells expressed AP-2alpha protein only in the cytoplasm. In carcinomas nuclear AP-2alpha expression was observed in 28% of the cases although cytoplasmic expression was more common (51%). The expression of AP-2alpha varied according to the histological subtype and differentiation. AP-2alpha and p21/WAF1 expressions did not correlate with each other. Both in univariate (P = 0.002) and multivariate analyses (relative risks (RR) 1.6, 95% confidence interval (CI) 1.13-2.18, P= 0.007) the high cytoplasmic AP-2alpha expression favoured the overall survival. In contrast, the nuclear AP-2alpha expression combined with low cytoplasmic expression increased the risk of dying of ovarian cancer (RR = 2.10, 95% CI 1.13-3.83, P= 0.018). The shift in the expression pattern of AP-2alpha (nuclear vs cytoplasmic) in carcinomas points out to the possibility that this transcription factor may be used by oncogenes in certain histological subtypes. Based on the mRNA analyses, the incomplete expression and translation of AP-2alpha in ovarian cancer may be due to post-transcriptional regulation.

Aged↗
PubMed2000

Transcription factor activator protein-2 is required for continued luteinizing hormone-releasing hormone expression in the forebrain of developing mice.

LHRH is the neuropeptide responsible for reproductive function. Prenatally, LHRH expression begins when neurons are in the olfactory pit and continues as these cells migrate into the brain. Thus, LHRH neurons maintain neuropeptide expression through very distinct environments. The regulatory interactions that control onset and continued expression of the LHRH phenotype are unknown. To begin to address this question primary LHRH neurons were removed from nasal explants at different ages. A complementary DNA (cDNA) subtraction screen was performed comparing a 3.5-days in vitro LHRH neuron [approximately embryonic day 15 (E15) in vivo] to two 10.5-days in vitro LHRH neurons (approximately postnatal day 1 in vivo). The transcription factor activator protein-2 (AP-2alpha) was differentially expressed and was present in the developmentally younger LHRH neuron. In vivo analysis revealed that LHRH neurons expressed AP-2 as they migrated across the cribriform plate and into the forebrain beginning on E13.5, but that coexpression of LHRH and AP-2 was no longer detected in postnatal day 1 animals. This suggested a regulatory role for AP-2 in LHRH neurons. Analysis of animals lacking AP-2alpha revealed a dramatic decrease in forebrain LHRH neurons between E13.5 and E14.5, correlating with normal onset of AP-2 expression in LHRH neurons as they entered the central nervous system. Nasal cells robustly expressing LHRH were still present on E 14.5. The continued presence of forebrain LHRH cells is proposed based on a second marker, galanin, and lack of increased apoptotic/necrotic cells in this region. A decrease in LHRH messenger RNA in forebrain neurons indicates regulation of LHRH occurred at the transcriptional or posttranscriptional level in mutant animals. These results indicate a developmentally restricted involvement of the transcription factor AP-2 in LHRH expression once the LHRH neurons have migrated into the forebrain, but before establishment of an adult-like distribution.

Animals↗
PubMed1999

Keratin 10 gene expression during differentiation of mouse epidermis requires transcription factors C/EBP and AP-2.

The epidermis forms a vital barrier composed of stratified keratinocytes and their differentiated products. One of these products, keratin K10, is critical to epidermal integrity, because mutations in k10 lead to abnormal blistering. For the normal expression of k10, differentiation-associated transcription factors C/EBPalpha, C/EBPbeta, and AP-2 are well positioned to play an important role. Here, regulation of the k10 gene is examined in keratinocytes in the skin of normal mice and in transgenic mice carrying targeted deletions of c/ebpbeta and ap-2alpha. In cultured cells, C/EBPalpha and C/EBPbeta are each capable of activating the k10 promoter via three binding sites, identified by site-directed mutagenesis. In a given epidermal cell in vivo, however, the selection of C/EBPalpha versus C/EBPbeta for k10 regulation is determined via a third transcription factor, AP-2. This novel regulatory scheme involves: (1) unique gradients of expression for each transcription factor, i.e., C/EBPbeta and AP-2 most abundant in the lower epidermis, C/EBPalpha in the upper; (2) C/EBP-binding sites in the ap-2alpha gene promoter, through which C/EBPbeta stimulates ap-2alpha; and (3) AP-2 binding sites in the c/ebpalpha promoter, through which AP-2 represses c/ebpalpha. Promoter-analysis and gene-expression data presented herein support a regulatory model in which C/EBPbeta activates and maintains AP-2 expression in basal keratinocytes, whereas AP-2 represses C/EBPalpha in those cells. In response to differentiation signals, loss of AP-2 expression leads to derepression of the c/ebpalpha promoter and activation of k10 as cells migrate upward.

Animals↗
PubMed1999

Osteoblast biocompatibility of mineral trioxide aggregate.

This study investigated the biocompatibility of variants of mineral trioxide aggregate (MTA), by culturing human MG63 osteosarcoma cells in the presence of materials, observing cytomorphology and cell growth, and then assaying cytokine expression from the cells. Reference materials were employed. Cell growth was quantified by preparing samples (n = 6) at 2, 4 and 7 days, for viewing by scanning electron microscopy and then scoring the amount of material that was covered by healthy cells. Subsequently, samples of culture media were tested using ELISA assays for expression of Interleukin (IL)-1alpha, IL-6, IL-8, IL-11 and macrophage colony stimulating factor (M-CSF). These assays were compared with controls where no material was present, and where media and fetal calf serum had not been exposed to cells. Results showed good cell growth on MTA. Expression of IL-6 from cells was only evident in the presence of MTA and Interpore 200. Interleukin-8 was expressed in high concentrations only in the presence of MTA. There was no evidence of expression of IL-1alpha or IL-11 with any material. Production of M-CSF was high for all materials. It appears that the variants of MTA are biocompatible and suitable for use in clinical trials.

Aluminum Compounds↗
PubMed1999

Diffusion-weighted magnetic resonance imaging of intracranial epidermoid tumours.

Intracranial epidermoids can closely mimic cerebrospinal fluid (CSF) on MRI and CT. Therefore epidermoids can be difficult to detect, or distinguish from CSF. Three cases of intracranial epidermoid are presented, one of which closely mimicked an arachnoid cyst on CT and routine MRI sequences. Diffusion-weighted magnetic resonance imaging (DWI) was performed. All three epidermoids demonstrated marked restriction of diffusion relative to CSF, clearly defining the extent of each lesion, and allowing differentiation from an arachnoid cyst or an enlarged CSF space.

Adult↗
PubMed1999

Comparison of two Doppler ultrasound criteria for grading cervical internal carotid artery stenosis.

In a review of 49 carotid arteries that had been assessed by ultrasound and angiography, two Doppler criteria were compared for accuracy in grading internal carotid artery (ICA) stenosis. The Seattle criteria for Doppler spectral analysis and the internal-to-common carotid artery peak systolic velocity ratio were of similar accuracy. When used in combination, there was an increase in sensitivity compared with the use of a single criterion. The velocity ratio had a higher sensitivity for detection of high-grade stenosis, but it tended to overestimate the grade of stenosis more than the Seattle criteria. To increase the sensitivity for detection of severe stenosis, the criteria should be used together and the higher grade of stenosis should be taken if there is a mismatch in assessment.

Carotid Artery, Common↗
PubMed1999

Treatment of infrarenal aortic stenosis by stent placement: a 6-year experience.

There is a significant re-stenosis rate with percutaneous treatment of stenoses of the infrarenal abdominal aorta with balloon angioplasty. Since 1990 the authors have primarily treated local infrarenal aortic stenoses with metallic endoluminal stents. The authors' experience with 12 consecutive patients (nine women and three men, aged from 30 to 72 years (mean age = 57 years)) is presented. Follow-up is available in 11 cases over 7-78 months (mean 32 months). The procedure was technically successful in all patients. Of the 11 patients with follow-up available, claudication was cured (n = 7) or significantly improved (n = 4). Those with persisting claudication had concurrent distal arterial disease. Periprocedural complications occurred in five cases, with two significant complications. One case required iliac angioplasty for embolized aortic atherosclerotic plaque, and one case required surgical thrombectomy and vein patch for iliac thrombosis complicating iliac dissection, without long-term sequelae in either case. One patient has had recurrent symptomatic aortic stenosis occurring 6 years after initial stenting, which responded to further stent insertion. Primary patency of 91% and secondary assisted patency of 100% has been achieved. Primary treatment of infrarenal aortic stenosis with endoluminal stenting results in high patency rates, with low morbidity and relatively low complication rates.

Adult↗
PubMed1999

Interventional catheter magnetic resonance angiography with a conventional 1.5-T magnet: work in progress.

Magnetic resonance contrast enhancement depends on the relative timing of image acquisition. Limited human trials have demonstrated efficacy of intra-arterial gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) in delineating vascular anatomy with X-rays. The present study assessed the ability of dynamic MR during intra-arterial Gd-DTPA administration to demonstrate vascular anatomy compared to conventional angiography as the gold standard. As interventional MR techniques using dedicated magnets proliferate, the ability to perform invasive MR angiography with a conventional magnet would be of great utility at established sites. Four subjects referred for different types of angiography underwent dynamic MR studies, including one with iliac artery stenting (Palmaz P204, Johnson and Johnson). All were examined with conventional angiography, and again after dynamic intra-arterial (IA) Gd-DTPA infusion. Coronal MRI images of the body were acquired using a 1.5-T superconducting magnet (three with a GE Signa, one with Philips NT), fast spoiled gradient echo (FSPGR); echo time (TE) = 4.2 msec, repetition time (TR) = 68-150 msec, flip = 75 degrees, 0-600 s after dilute Gd-DTPA IA bolus injection during sequential breath-hold acquisitions of 13-32 s each. All arteries were detected with dynamic MR. The FSPGR MRI with IA Gd-DTPA administration can provide adequate time and spatial resolution to demonstrate arterial anatomy and arterial stent patency.

Carotid Artery Diseases↗