Biomedical subjects

E Mueller

Publications and source records attributed to E Mueller.

At least 19 recordsLinked to original sources

PubMed2005

Chromosomal instability in flat adenomas and carcinomas of the colon.

Flat adenomas are flat or slightly elevated dysplastic lesions of the colorectal mucosa, mostly with a tubular architecture. Compared with polypoid adenomas of similar size, flat adenomas show a higher frequency of high-grade dysplasia and rapid submucosal invasion. The aim of this study was to survey whether flat colorectal lesions differ in their pattern of chromosomal aberrations from their polypoid counterparts. Six flat adenomas and 12 flat carcinomas were analysed by comparative genomic hybridization (CGH) and the pattern of chromosomal aberrations was compared with a previously published series of 112 polypoid adenomas and 82 polypoid carcinomas. In addition, multiplex ligation-dependent probe amplification (MLPA) for identifying DNA copy number changes of 25 individual genes on chromosome 20 was performed on 14 flat and 15 polypoid tumours. With CGH, flat adenomas showed on average 1.8 gains (range 1-4) and 3.2 losses (range 0-4), and the flat carcinomas 4.5 gains (range 0-8) and 3.5 losses (range 1-6). In both adenomas and carcinomas, high frequencies of 20q gain (83% and 92%, respectively) and 18q loss (83% and 92%, respectively) were found. This correlation between 20q gain and 18q loss had previously been observed in a subgroup of polypoid colorectal tumours. Both flat and polypoid colorectal tumours with 20q gains by CGH showed similar patterns of copy number ratios for the individual genes tested. TOP1, BCL2L1, and E2F1 had median copy number ratios of 2 or higher, while ZNF217 had a ratio around 3. In conclusion, flat adenomas and carcinomas of the large intestine show a similar pattern of chromosomal aberrations to that observed in a specific subgroup of polypoid lesions. The transcription factor ZNF217 is an important candidate for driving the 20q gain.

Adenoma↗

PubMed2005

Gender differences in 24-hour urinary diaries of asymptomatic North American adults.

PURPOSE: Two previously published studies from our center have described the urinary habits of asymptomatic men (284) and women (300) as revealed by 24-hour urinary diaries. Those gender specific studies found that urinary diary variables are affected by age and race. By comparing the data from those studies we determined the effect of gender on voiding habits. MATERIALS AND METHODS: In this secondary analysis we matched each female urinary diary to that of a male of similar age and race. Diary variables were compared using paired sign tests with results considered significant at the 5% level. RESULTS: A total of 141 matched pairs were studied. The population age ranged from 18 to 68 years and was racially diverse (56% black, 31% white, 7% Hispanic and 6% Asian). Men had higher total fluid intake and mean voided volume than women (p <0.001 and 0.04, respectively). Women voided more frequently than men (p = 0.006) and had more voids per liter of fluid intake (p <0.001). No gender differences were found for body mass index, nighttime or daytime diuresis rates, total urine volume, maximum voided volume or rates of nocturia. CONCLUSIONS: This analysis suggests that there are significant gender effects on 24-hour diary variables, with females tending to void more often and at lower mean volumes. The results of our study may be useful in the design of research studies or for patient counseling.

Adolescent↗

PubMed2004

Education in the broader sense.

Education is the key element in medical science. It is generally divided into pre-graduate, postgraduate and continuing medical education, with the physician as the main target. It usually covers aetiology, diagnosis, treatment, prognosis, outcome and long-term effects. Neurology is a large field in medical science, focusing on diseases of the brain, the spinal cord, the peripheral nerves and muscles. They tend to be chronic and, often progressive, leading to long-term disability and handicap for the individual, frequently associated with personal, social and emotional consequences. In the time span of a patient with a neurological disease, interaction with physicians is important and often vital, but interaction with other health groups and relatives or other caregivers is longer and more intensive. The knowledge and experience of the patient and caregiver is equally important, in that an informed patient or caregiver is more likely to participate actively and be of greater benefit in disease management. This paper promotes the concept that education in neurology should be integrated to include the physicians and other professionals as well as patients, caregivers and public. A broader awareness of neurological diseases will draw support and improve the approach of society to the provision of resources and management of these illnesses, as well as public attitude towards the symptoms and to those who live with them. Two bodies have collaborated in writing this article: the education committee of the EFNS and the EFNA.

Caregivers↗

PubMed2001

A comparative study of E-cadherin and stromelysin-3 expression in de novo and ex adenoma carcinoma of the colorectum.

An apparent exception to the colorectal adenoma-carcinoma carcinogenetic pathway is the so-called "de novo" carcinoma which has no evidence of adenoma in its vicinity. Despite the fact that they are often quite small, these lesions appear to be more aggressive (i.e., greater likelihood of lymph-node metastases) than carcinomas that clearly arise from surrounding adenomas exadenoma carcinoma. The purpose of the present comparative immunohistochemical study was to compare rates of cell adhesion molecule (E-cadherin) and protease [stromelysin-3 (ST-3)] expression in groups of de novo (n=64) and ex adenoma (n=42) lesions in order to see whether their more aggressive behavior is associated with decreased cell adhesion and increased protease expression. The rates of extensive ST-3 expression and decreased E-cadherin expression were significantly higher in the de novo group (P=0.014 and 0.005, respectively). Histopathologically, the de novo group also had a significantly higher percentage of cases with an infiltrative invasion pattern. These differences highlight the more aggressive phenotype of the de novo colorectal carcinoma and fit with their greater invasive potential.

Adenoma↗

PubMed2001

Characterization of the intracellular survival of Mycobacterium avium ssp. paratuberculosis: phagosomal pH and fusogenicity in J774 macrophages compared with other mycobacteria.

The phagosomes containing viable pathogenic mycobacteria, such as Mycobacterium (M.) tuberculosis and Mycobacterium avium ssp. avium (M. avium), are known to be limited in their ability to both acidify and fuse with late (but not early) endocytic organelles. Here, we analysed the pH and fusogenicity of phagosomes containing M. avium ssp. paratuberculosis (M. ptb), the causative agent of paratuberculosis in ruminants. Using the murine J774 macrophage cell line, we compared viable and heat-killed M. ptb and, in addition, viable or dead M. avium, as well as two non-pathogenic mycobacteria, Mycobacterium smegmatis and Mycobacterium gordonae. Electron microscopic analysis revealed that M. ptb persisted intracellularly in phagosomes for up to 15 days. The phagosomes containing live M. ptb and M. avium were significantly reduced in their ability to acquire some markers for the endocytic pathway, such as internalized calcein, BSA-gold or the membrane protein Lamp 2. However, they were almost completely accessible to 70 kDa fluorescein isothiocyanate (FITC)-dextran and Lamp 1. Overall, the phagosomes containing dead pathogenic mycobacteria behaved similarly to the ones containing live non-pathogenic mycobacteria in all experiments. Using FITC-dextran in a novel fluorescence-activated cell sorting (FACS)-based method, we could also show that the bulk of endocytic compartments, including phagosomes, were only very mildly acidified to approximately pH 6.3 over at least 72 h in J774 cells infected with live M. ptb and M. avium. In contrast, J774 cells treated with heat-killed M. ptb or BSA-coated latex beads showed substantial acidification of the phagosome/endocytic compartments to a pH value of approximately 5.2. After infection with M. smegmatis and M. gordonae, acidification was initially (1-5 h after infection) inhibited, but increased after longer infection to levels similar to those with dead mycobacteria.

Animals↗

PubMed2001

Ornithine decarboxylase knockout in Tapesia yallundae abolishes infection plaque formation in vitro but does not reduce virulence toward wheat.

A knockout strain of Tapesia yallundae lacking the single ornithine decarboxylase (ODC) allele has been created by targeted gene replacement. A central region of the ODC gene was isolated by polymerase chain reaction with degenerate oligonucleotides and used to probe a lambda genomic library. The gene was sequenced, and the encoded ODC protein sequence was shown to be similar to those from other fungi. The functionality of the T. yallundae ODC was confirmed by complementation of an Aspergillus nidulans mutant (puA) strain devoid of ODC activity, restoring growth in the absence of exogenous polyamines. Transformation-mediated gene replacement was used to create strains that were auxotrophic for putrescine and lack ODC coding sequences. ODC knockout strains were unable to differentiate infection structures after in vitro induction and showed an abnormal hyphal branching phenotype. Pathogenicity studies on these mutants showed that, surprisingly, they are not reduced in virulence compared with nondisrupted transformants. This suggests that the strains carrying an ODC disruption can obtain sufficient polyamines from the host plant for normal growth and differentiation and, therefore, that fungal ODC may not be a suitable target for fungicides.

Ascomycota↗

PubMed2001

Use of a hyaluronate membrane for jejunal anastomosis in horses.

OBJECTIVE: To compare the outcomes of double-layer inverting anastomosis (DIA), single-layer anastomosis (SLA), and single-layer anastomosis combined with a hyaluronate membrane (SLA+HA-membrane) with respect to stomal diameter, adhesion formation, surgery time, and anastomotic healing in horses. ANIMALS: 18 adult horses. PROCEDURE: Midline celiotomy and end-to-end anastomoses were performed. In control horses (n = 6), DIA was performed; in treated horses, SLA was performed (6) or SLA+HA-membrane was performed (6). Horses were euthanatized 21 days after surgery. Abdominal adhesions were evaluated grossly and histologically. Stomal diameters were measured ultrasonographically and compared with adjacent luminal diameters. Anastomotic healing was evaluated histologically for fibrosis and inflammation, tissue alignment, and inversion. Surgery times were recorded for the anastomotic procedure and compared among groups. RESULTS: There were significantly more adhesions in the SLA group, compared with the DIA and SLA+HA-membrane groups. Reduction in stomal diameters in the DIA group was significantly greater than the SLA and SLA+HA-membrane groups. Surgery times for the DIA group were significantly greater than the SLA and SLA+HA-membrane groups. Histologic findings of fibrosis, inflammation, and mucosal healing were similar among groups. There was significant tissue inversion in the DIA group, compared with the 2 treatment groups. Tissue alignment was not different among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Use of a SLA+HA-membrane was an effective small intestinal anastomotic technique. This technique was faster to perform and resulted in a larger stomal diameter, compared with the DIA technique and significantly fewer perianastomotic adhesions, compared with the SLA technique.

Anastomosis, Surgical↗

PubMed2001

Expression of the fibroblast activation protein during mouse embryo development.

Human Fibroblast Activation Protein (FAP), a member of the serine prolyl oligopeptidase family, is a type II cell surface glycoprotein that acts as a dual-specificity dipeptidyl-peptidase (DPP) and collagenase in vitro. Its restricted expression pattern in embryonic mesenchyme, in wound healing and in reactive stromal fibroblasts of epithelial cancers, has suggested a role for the FAP protease in extracellular matrix degradation or growth factor activation in sites of tissue remodeling. The FAP homologue in Xenopus laevis has been reported to be induced in the thyroid hormone-induced tail resorption program during tadpole metamorphosis supporting a role for FAP in tissue remodeling processes during embryonic development. However, Fap-deficient mice show no overt developmental defects and are viable. To study the expression of FAP during mouse embryogenesis, a second Fap-deficient mouse strain expressing beta-Galactosidase under the control of the Fap promoter was generated by homologous recombination (Fap-/- lacZ mice). FAP deficiency was confirmed by the absence of FAP-specific dipeptidyl-peptidase activity in detergent-soluble extracts isolated from 17.5 d.p.c. Fap-/- lacZ embryos. We report that Fap-/- lacZ mice express beta-Galactosidase at regions of active tissue remodeling during embryogenesis including somites and perichondrial mesenchyme from cartilage primordia.

Animals↗

PubMed2000

Effects of ligand activation of peroxisome proliferator-activated receptor gamma in human prostate cancer.

Peroxisome proliferator-activated receptor gamma (PPARgamma) is a nuclear hormone receptor that plays a key role in the differentiation of adipocytes. Activation of this receptor in liposarcomas and breast and colon cancer cells also induces cell growth inhibition and differentiation. In the present study, we show that PPARgamma is expressed in human prostate adenocarcinomas and cell lines derived from these tumors. Activation of this receptor with specific ligands exerts an inhibitory effect on the growth of prostate cancer cell lines. Further, we show that prostate cancer and cell lines do not have intragenic mutations in the PPARgamma gene, although 40% of the informative tumors have hemizygous deletions of this gene. Based on our preclinical data, we conducted a phase II clinical study in patients with advanced prostate cancer using troglitazone, a PPARgamma ligand used for the treatment of type 2 diabetes. Forty-one men with histologically confirmed prostate cancer and no symptomatic metastatic disease were treated orally with troglitazone. An unexpectedly high incidence of prolonged stabilization of prostate-specific antigen was seen in patients treated with troglitazone. In addition, one patient had a dramatic decrease in serum prostate-specific antigen to nearly undetectable levels. These data suggest that PPARgamma may serve as a biological modifier in human prostate cancer and its therapeutic potential in this disease should be further investigated.

Aged↗

PubMed2000

PAX8-PPARgamma1 fusion oncogene in human thyroid carcinoma [corrected].

Chromosomal translocations that encode fusion oncoproteins have been observed consistently in leukemias/lymphomas and sarcomas but not in carcinomas, the most common human cancers. Here, we report that t(2;3)(q13;p25), a translocation identified in a subset of human thyroid follicular carcinomas, results in fusion of the DNA binding domains of the thyroid transcription factor PAX8 to domains A to F of the peroxisome proliferator-activated receptor (PPAR) gamma1. PAX8-PPARgamma1 mRNA and protein were detected in 5 of 8 thyroid follicular carcinomas but not in 20 follicular adenomas, 10 papillary carcinomas, or 10 multinodular hyperplasias. PAX8-PPARgamma1 inhibited thiazolidinedione-induced transactivation by PPARgamma1 in a dominant negative manner. The experiments demonstrate an oncogenic role for PPARgamma and suggest that PAX8-PPARgamma1 may be useful in the diagnosis and treatment of thyroid carcinoma.

Adenocarcinoma, Follicular↗

PubMed2000

Degradation of the peroxisome proliferator-activated receptor gamma is linked to ligand-dependent activation.

The nuclear hormone receptor peroxisome proliferator-activated receptor (PPAR) gamma is a ligand-activated transcription factor that regulates several crucial biological processes such as adipogenesis, glucose homeostasis, and cell growth. It is also the functional receptor for a new class of insulin-sensitizing drugs, the thiazolidinediones, now widely used in the treatment of type 2 diabetes mellitus. Here we report that PPARgamma protein levels are significantly reduced in adipose cells and fibroblasts in response to specific ligands such as thiazolidinediones. Studies with several doses of different ligands illustrate that degradation of PPARgamma correlates well with the ability of ligands to activate this receptor. However, analyses of PPARgamma mutants show that, although degradation does not strictly depend on the transcriptional activity of the receptor, it is dependent upon the ligand-gated activation function 2 (AF2) domain. Proteasome inhibitors inhibited the down-regulation of PPARgamma and ligand activation enhanced the ubiquitination of this receptor. These data indicate that, although ligand binding and activation of the AF2 domain increase the transcriptional function of PPARgamma, these same processes also induce ubiquitination and subsequent degradation of this receptor by the proteasome.

3T3 Cells↗

PubMed2000

Ornithine decarboxylase of Stagonospora (Septoria) nodorum is required for virulence toward wheat.

A knockout strain of Stagonospora (Septoria) nodorum lacking the single ornithine decarboxylase (ODC) allele has been created by targeted gene replacement. A central region of the S. nodorum ODC gene was isolated by polymerase chain reaction using degenerate oligonucleotides and used to probe a lambda genomic library. The gene was sequenced and the encoded ODC protein sequence was shown to be similar to those from other fungi. The functionality of the S. nodorum ODC was confirmed by complementation of an Aspergillus nidulans mutant (puA) strain devoid of ODC activity, restoring growth in the absence of exogenous polyamines. Sporulation of the transformants was reduced suggesting abberant regulation of the S. nodorum gene in A. nidulans. Transformation-mediated gene replacement was used to create strains which were auxotrophic for putrescine and lack ODC coding sequences. Pathogenicity studies on these mutants showed that they are greatly reduced in virulence compared with non-disrupted transformants. This confirms that the strains carrying an ODC disruption cannot obtain sufficient polyamines from the host plant for normal growth and, thus, that fungal ODC may be a suitable target for chemical intervention.

Amino Acid Sequence↗

PubMed2000

Prospective acquisition correction for head motion with image-based tracking for real-time fMRI.

In functional magnetic resonance imaging (fMRI) head motion can corrupt the signal changes induced by brain activation. This paper describes a novel technique called Prospective Acquisition CorrEction (PACE) for reducing motion-induced effects on magnetization history. Full three-dimensional rigid body estimation of head movement is obtained by image-based motion detection to a high level of accuracy. Adjustment of slice position and orientation, as well as regridding of residual volume to volume motion, is performed in real-time during data acquisition. Phantom experiments demonstrate a high level of consistency (translation < 40 microm; rotation < 0.05 degrees ) for detected motion parameters. In vivo experiments were carried out and they showed a significant decrease of variance between successively acquired datasets compared to retrospective correction algorithms.

Algorithms↗

PubMed2000

Influence of hypothermia on cell volume and cytotoxic swelling of glial cells in vitro.

In view of the increasing significance of mild hypothermia (32 degrees C) as an efficient procedure of neuroprotection, the present study was performed to examine the influence of this level of hypothermia on the volume of glial cells under physiological as well as under pathological conditions. The influence of mild (32 degrees C) and moderate (27 degrees C) hypothermia on cell volume and cell viability of C6 glioma cells was studied for 60 minutes in vitro. Cells were suspended in an incubation chamber under continuous control of temperature, pH and pO2. Cell volume was measured by an advanced Coulter system. Hypothermia itself was causing significant cell swelling in a dose-dependent manner, which could be prevented by omission of Na(+)-ions from the suspension medium, while the replacement of Cl(-)-ions failed to prevent cell swelling from hypothermia. Inhibition of the Na+/H(+)-antiporter with EIPA (5N-ethyl-n-isopropyl-amiloride, 50 microM) was significantly reducing the hypothermia induced cell swelling, indicating activation of the Na+/H(+)-antiporter. Conversely, mild or moderate hypothermia failed to prevent cell swelling from lactic acid, arachidonic acid or glutamate, i.e. agents which are mediating the development of cytotoxic brain edema in vivo in cerebral trauma, ischemia and other acute insults. The findings indicate that cerebral protection by hypothermia in vivo is most likely not attributable to an inhibition of cytotoxic brain edema. Further investigations, however, are required in vivo and in vitro to elucidate the hypothermia-induced swelling of glial cells in more detail, e.g. as to the role of the Na+/H(+)-antiporter.

Brain Neoplasms↗

PubMed2000

A molecular biomarker system for assessing the health of coral (Montastraea faveolata) during heat stress.

Using a novel molecular biomaker system (MBS), we assessed the physiological status of coral (Montastraea faveolata) challenged by heat stress by assaying specific cellular and molecular parameters. This technology is particularly relevant for corals because heat stress is thought to be an essential component of coral bleaching. This phenomenon is widely believed to be responsible for coral mortality worldwide, particularly during 1997-1998. Specific parameters of coral cellular physiology were assayed using the MBS that are indicative of a nonstressed or stressed condition. The MBS distinguished the separate and combined effects of heat and light on the 2 coral symbionts, a scleractinian coral and a dinoflagellate algae (zooxanthellae). This technology aids in the accurate diagnosis of coral condition because each parameter is physiologically well understood. Finally, the MBS technology is relatively inexpensive, easy to implement, and precise, and it can be quickly adapted to a high-throughout robotic system for mass sample analysis.

Journal Article↗

PubMed2000

Potato virus X amplicons in arabidopsis mediate genetic and epigenetic gene silencing.

Amplicon transgenes from potato virus X (PVX) are based on a modified version of the viral genome and are efficient activators of post-transcriptional gene silencing (PTGS). To determine whether PVX amplicons activate PTGS in Arabidopsis, we used constructs based on the genome of PVX carrying a green fluorescent protein (GFP) reporter gene. Our analysis of the transgene phenotype exploited previous observations indicating that PTGS is associated with short 25-nucleotide RNA species, transgene methylation, and homology-dependent virus resistance. We also used the ability of turnip mosaic virus to suppress gene silencing as a means of dissecting stages of the mechanism. The results showed that a PVX:GFP amplicon induces weak PTGS and that this PTGS was enhanced in the presence of a GFP reporter gene. Our interpretation of these data is that the PTGS induced by the amplicon was genetically determined and equivalent to the initiation stage of the PTGS mechanism. The PTGS induced by the combined amplicon and reporter gene was equivalent to the maintenance stage and was associated with an epigenetic conversion of the transgene. The distinction between genetic and epigenetic PTGS explains the well-characterized effects of transgene dosage on PTGS that have been previously interpreted in terms of RNA expression thresholds.

Arabidopsis↗

PubMed2000

Over-representation of PPARgamma sequence variants in sporadic cases of glioblastoma multiforme: preliminary evidence for common low penetrance modifiers for brain tumour risk in the general population.

PPARgamma, the gamma isoform of a family of peroxisome proliferator activated receptors, plays a key role in adipocyte differentiation. Recently, its broad expression in multiple tissues and several epithelial cancers has been shown. Further, somatic loss of function mutations in PPARgamma have been found in primary colorectal carcinomas. We sought to determine if somatic high penetrance mutations in this gene might also play a role in glioblastoma multiforme (GBM). We also examined this gene to determine if common low penetrance polymorphic alleles might lend low level susceptibility to GBM in the general population. No somatic high penetrance mutations were detected in 96 sporadic GBMs. However, polymorphic alleles at codons 12 and 449 were significantly over-represented among the 27 unrelated American patients with sporadic GBM compared to 80 race matched controls. While nine (33%) were heterozygous for the P12A variant, c.34C/G (cytosine to guanine change at nucleotide 34), 12 (15%) controls were heterozygous for P12A (p<0.05). Similarly, 13 of 26 (50%) glioblastoma patients compared to 10 of 80 (12%) normal controls were found to have the heterozygous H449H polymorphism (p<0.001). The over-representation of H449H in glioblastoma patients was confirmed with a second validation set of American patients. When both American series were combined, polymorphic H449H was over-represented among cases versus controls (p<0.001) and there was a similar trend (p=0.07) for P12A. The precise mechanism for this association is unknown but these PPARgamma polymorphisms may be acting in a low penetrance predisposing manner. However, these associations were not found in a German population, possibly arguing that if these variants are in linkage disequilibrium with a third locus, then this effect is relatively new, after the settlement of the American colonies.

Alleles↗

PubMed2000

Expression of secretory group IIA phospholipase A(2) in relation to the presence of microbial agents, macrophage infiltrates, and transcripts of proinflammatory cytokines in human aortic tissues.

Recent seroepidemiological and immunohistochemical studies have demonstrated an association between microbial infections and atherosclerosis. However, the mechanisms underlying this association are widely unknown. In the present study, arterial specimens obtained at autopsy after sudden death were analyzed concerning (1) the presence of Chlamydia pneumoniae, cytomegalovirus, herpes simplex virus, and Helicobacter pylori; (2) the expression of secretory group IIA phospholipase A(2) (sPLA(2)-IIA) and of proinflammatory cytokines; and (3) the stage of atherosclerosis. Genomic DNA of microbial pathogens was determined by the polymerase chain reaction technique. The expression of sPLA(2)-IIA was studied immunohistochemically by using monoclonal antibodies against human sPLA(2)-IIA. Transcripts specific for sPLA(2)-IIA, interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma were identified by reverse transcription-polymerase chain reaction. In 18 of 102 analyzed specimens, DNA of microbial pathogens was found. Thirteen sections were positive for C pneumoniae, whereas 2 specimens were positive either for cytomegalovirus or for herpes simplex virus. One section contained genomic DNA of all 3 pathogens simultaneously. None of the analyzed tissues exhibited nucleic acids specific for H pylori. In addition to macrophage infiltrates, the presence of microbial DNA was closely associated with the occurrence of transcripts specific for proinflammatory cytokines and sPLA(2)-IIA. Pathogens as well as sPLA(2)-IIA and cytokines were found to be present not only in advanced but also in early stages of atherosclerosis. In tissues negative for sPLA(2)-IIA and cytokine expression, none of the pathogens could be identified. Because macrophages exposed to phospholipase A(2)-treated lipoproteins are transformed into foam cells in vitro, the results of this study suggest an alternative mechanism by which microbial infections may act in a proatherogenic fashion in vessel walls.

Adolescent↗