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Targeted cellular process profiling approach for uterine leiomyoma using cDNA microarray, proteomics and gene ontology analysis.

This study utilized both cDNA microarray and two-dimensional protein gel electrophoresis technology to investigate the multiple interactions of genes and proteins involved in uterine leiomyoma pathophysiology. Also, the gene ontology analysis was used to systematically characterize the global expression profiles at cellular process levels. We profiled differentially expressed transcriptome and proteome in six-paired leiomyoma and normal myometrium. Screening up to 17 000 genes identified 21 upregulated and 50 downregulated genes. The gene-expression profiles were classified into mutually dependent 420 functional sets, resulting in 611 cellular processes according to the gene ontology. Also, protein analysis using two-dimensional gel electrophoresis identified 33 proteins (17 upregulated and 16 downregulated) of more than 500 total spots, which was classified into 302 cellular processes. Of these functional profilings, downregulations of transcriptomes and proteoms were shown in cell adhesion, cell motility, organogenesis, enzyme regulator, structural molecule activity and response to external stimulus functional activities that are supposed to play important roles in pathophysiology. In contrast, the upregulation was only shown in nucleic acid-binding activity. Taken together, potentially significant pathogenetic cellular processes were identified and showed that the downregulated functional profiling has a significant impact on the discovery of pathogenic pathway in leiomyoma. Also, the gene ontology analysis can overcome the complexity of expression profiles of cDNA microarray and two-dimensional protein analysis via its cellular process-level approach. Therefore, a valuable prognostic candidate gene with relevance to disease-specific pathogenesis can be found at cellular process levels.

Cell Communication↗

Relationships between insulin-like growth factor-1 levels and growth hormone concentrations during diurnal profiles and following oral glucose in acromegaly.

OBJECTIVE: The aim of this study was to refine the biochemical definition of disease activity in acromegaly by comparing serum growth hormone (GH) measurements during a 10-hour day profile with serum GH values during an oral glucose tolerance test. DESIGN: Using plasma insulin-like growth factor-1 (IGF-1) levels as a measure of disease activity, serum GH data from a day profile and from an oral glucose tolerance test were compared. PATIENTS: Thirty-five acromegalic patients were studied, 13 of whom had serum GH measured during a day profile and 22 during an oral glucose tolerance test. In addition, basal plasma IGF-1 levels were estimated in all acromegalic patients, and in 24 normal subjects. MEASUREMENTS: Following acid-ethanol extraction of the plasma samples, IGF-1 levels were measured by radioimmunoassay using a polyclonal antibody. In a day profile, six to eight blood samples for serum GH estimation were taken at hourly intervals during the day; during an oral glucose tolerance test samples for serum GH estimation were taken in the fasting state and every 30 minutes for 2 hours and measured by a two-site IRMA for GH. RESULTS: Ninety-four per cent of acromegalic patients with raised plasma IGF-1 levels had serum GH concentrations > 10 mU/l whilst 98% of acromegalic patients with plasma IGF-1 levels in the normal range had serum GH concentrations < 6 mU/l. A highly significant positive correlation was found between the mean serum GH concentrations (r = 0.67), the minimum serum GH concentration (r = 0.65) and the area under the GH curve (r = 0.66) estimated during an oral glucose tolerance test and plasma IGF-1 concentrations. The relations between identical indices of serum GH concentration measured during a day profile and plasma IGF-1 levels, although significant, show a less powerful correlation. The relation between serum GH and plasma IGF-1 levels describes a curvilinear model, plasma IGF-1 levels exhibiting a plateau at serum GH concentrations > 40 mU/l but maintaining a linear relationship with serum GH levels < 20 mU/l. CONCLUSIONS: A highly significant correlation exists between plasma IGF-1 levels and various parameters of serum GH levels in acromegalic patients. Hormonal assessment of disease activity in acromegaly is more accurately reflected by the serum GH concentration during an oral glucose tolerance test rather than by the serum GH level during a day profile. Normalization of plasma IGF-1 levels is rarely achieved unless the mean serum GH level is reduced to < 6 mU/l.

Acromegaly↗

Use of plasmid profiles to detect changes in strains of Staphylococcus aureus during poultry processing.

The plasmid profiles of Staphylococcus aureus strains isolated at different stages in three poultry processing plants have been examined. Changes in profiles were seen in two plants after the plucking stage and the appearance of these new profiles correlated with the presence of an endemic strain, as suggested previously by increases in bacterial counts and changes in biotypes at the same stage. A third plant in which such changes did not occur showed no change in profiles. Plasmid profiles are therefore a rapid and sensitive method for distinguishing endemic strains within a plant from the flora of the incoming birds. Certain profiles also appeared to correspond with particular biotypes and certain phage types.

Ampicillin Resistance↗

Geographical variation in antibiotic resistance profiles of Escherichia coli isolated from swine, poultry, beef and dairy cattle farm water retention ponds in Florida.

AIMS: The aim of this study was to assess geographical variation in multiple antibiotic resistance (MAR) profiles of livestock Escherichia coli as well as to evaluate the ability of MAR profiles to differentiate sources of faecal pollution. METHODS AND RESULTS: More than 2000 E. coli isolates were collected from water retention ponds and manure of swine, poultry, beef and dairy farms in south, central and north Florida, and analysed for MAR using nine antibiotics. There were significant differences in antibiotic resistance of E. coli by season and livestock type for more than one antibiotic, but regional differences were significant only for ampicillin. Over the three regions, discriminant analysis using MAR profiles correctly classified 27% of swine, 49% of poultry, 56% of beef and 51% of dairy isolates. CONCLUSIONS: Regional variations in MAR combined with moderate discrimination success suggest that MAR profiles of E. coli may only be marginally successful in identifying sources of faecal pollution. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the existence of regional and seasonal differences in MAR profiles as well as the limited ability of MAR profiles to discriminate among livestock sources.

Agriculture↗

Image analysis of Nissl-stained neuronal perikarya in the primary visual cortex of the rat: automatic detection and segmentation of neuronal profiles with nuclei and nucleoli.

An image analysing procedure for the morphometric characterization of cortical neurons in Nissl-stained brain sections is described. It consists of the automatic detection of cellular profiles and their compartments: cytoplasm, nucleus and nucleolus. The algorithm was designed to cope with the large morphological spectrum of cortical perikarya (e.g. geometrical properties of perikarya, staining intensities of cell compartments and nucleo-plasmic area-ratio) including pyramidal (Golgi-category I) and non-pyramidal (Golgi-category II) neurons. Clusters of cells were separated and non-neuronal structures (e.g. glia, endothelial cells) as well as tangential, non-nucleolated sections through neuronal perikarya recognized and excluded from further analysis without requiring interactive procedures. The performance of the profile recognition procedure was evaluated using 426 nucleolated and non-nucleolated profiles of different types of neurons in the primary visual cortex of the rat. Nucleolated profiles were recognized as such with a 91% accuracy, non-nucleolated profiles were rejected correctly in 90% of cases. After automatic segmentation and selection of nucleolated neuronal profiles from the microscopic field, a large set of quantitative morphological features including geometrical, densitometrical and textural parameters can be measured using high power light microscopy. This permits quantitative morphometric characterization of different neuronal types. This procedure is the first part of a system for the automatic classification of Nissl-stained cortical neurons.

Algorithms↗

Intranuclear membranous profiles in germinoma cells--a variant of nuclear pockets and intranuclear annulate lamellae.

When ultrastructurally examining 24 germinomas comprising 12 seminomas, 4 dysgerminomas, 1 mediastinal germinoma, and 7 intracranial germinomas, intranuclear membranous profiles were noticed in 17 germinomas, ranging from 20-100 nm in width and 3 microns in length. With occasional connections to the nuclear envelope through a small hole, intranuclear membranous profiles in germinoma cells were considered as clefts of the nuclear envelope. While most frequently situated under the inner nuclear membrane, they varied in configuration as well as distribution. As sequestered round mass of nuclear material, they were a variant of nuclear pockets containing nucleoplasm. Intranuclear annulate lamellae were occasionally present apart from the nuclear envelope and connected with nuclear clefts. Eleven of the twelve seminomas and 6 of the twelve non-seminomatous germinomas showed intranuclear membranous profiles, and the incidence of such profiles was much higher in seminomas than in non-seminomatous germinomas. Intranuclear membranous profiles facing the inner nuclear membrane were also noted in spermatogonia in adolescents and adults. It was suggested that intranuclear membranous profiles in germinoma cells could be structures following ones occasionally seen in spermatogonia.

Adolescent↗

Non-invasive estimation of intraurethral pressure profile from uroflowmetric curve.

BACKGROUND: An intraurethral pressure-time profile as urodynamic information was obtained in a non-invasive manner using an equivalent equation as a voiding model. METHODS: The reasonability of the voiding model was confirmed by applying it to an experimental flow curve likened to urinary flow. The flow curve was approximated and the pressure profile was estimated. From the uroflowmetric curves obtained in a normal subject and a patient with bladder outlet obstruction, the respective intraurethral pressure profiles were estimated. RESULTS: The pressure profile estimated from the approximated flow curve was found consistent with the profile of the difference between the pressure actually measured at two different portions in the experimental system. CONCLUSION: Non-invasive estimation of intraurethral pressure profile from uroflowmetric curves may be very useful to grasp intraurethral urodynamic information in clinical practice.

Models, Biological↗

Cognitive-behavioral profiles among different categories of orofacial pain patients: diagnostic and treatment implications.

Psychological homogeneity in temporomandibular disorders (TMD) is not conclusive. The multidimensional pain inventory (MPI) has previously identified 3 cognitive-behavioral profiles in TMD and chronic pain patients. Our aims were to replicate these findings in another cultural setting and relate the profiles to the diagnosis and to the treatment demand and outcome. The MPI was administered to 112 referrals comprising 6 categories of patients diagnosed with TMD or intractable orofacial pain. Dysfunctional profiles (high in pain and distress) were most common in patients with orofacial pain of obscure origin and more common in myofascial pain patients than in patients with other TMD diagnoses. Interpersonally-distressed profiles were found in all categories. Among patients with disk displacement, the 3rd profile (adaptive copers with low pain and distress and high control and activity) was most common in earlier successfully diskectomized patients and least common in those about to undergo invasive interventions. A dysfunctional profile was associated with treatment failure, conservative or surgical, and with the demand for radical therapy. Some support for a cyclical causality between pain and psychological factors was found. It is concluded that the robustness of the MPI as a relevant assessment instrument was further strengthened.

Adaptation, Psychological↗

Convex curvatures of alkaline elution profiles of DNA from human cells irradiated with 405 nm UVA: evidence for induction of slowly developing alkali-labile sites.

The alkaline (pH 12.1) elution profiles of DNA from human P3 cells exposed to monochromatic 405 nm UVA radiation deviate from exponential: on a logarithmic plot of eluted fraction of DNA vs time of elution, the rate of elution accelerates for the first 6 h. Following this period, the profiles become exponential. In contrast, the elution profiles of DNA after 520 nm green light or ionizing radiation exposures (x- and gamma rays, and fission spectrum neutrons) are always strictly exponential, evidence that the convex profiles were not due to an artifact caused by elution technique. Holding the DNA at pH 12.1 for 6 h after 405-nm exposures before initiating elution resulted in profiles that were close to exponential, with slopes similar to the final slopes observed following the 6-h elution period in the original experiments. This is evidence that some DNA breaks develop slowly during the first 6 h of elution, as a result of exposure to alkali. Therefore, the DNA lesions induced by 405-nm light as measured by the alkaline elution technique are apparently heterogeneous and include a major class of alkali-labile sites that develop slowly during incubation at pH 12.1. Convex profiles also occur following exposure of the cells to visible light at 434 and 512 nm.

DNA Damage↗

A pharmacological profile of the aldehyde receptor repertoire in rat olfactory epithelium.

Several lines of evidence suggest that odorants are recognized through a combinatorial process in the olfactory system; a single odorant is recognized by multiple receptors and multiple odorants are recognized by the same receptor. However few details of how this might actually function for any particular odour set or receptor family are available. Approaching the problem from the ligands rather than the receptors, we used the response to a common odorant, octanal, as the basis for defining multiple receptor profiles. Octanal and other aldehydes induce large EOG responses in the rodent olfactory epithelium, suggesting that these compounds activate a large number of odour receptors (ORs). Here, we have determined and compared the pharmacological profile of different octanal receptors using Ca(2+) imaging in isolated olfactory sensory neurones (OSNs). It is believed that each OSN expresses only one receptor, thus the response profile of each cell corresponds to the pharmacological profile of one particular receptor. We stimulated the cells with a panel of nine odorants, which included octanal, octanoic acid, octanol and cinnamaldehyde among others (all at 30microM). Cluster analysis revealed several distinct pharmacological profiles for cells that were all sensitive to octanal. Some receptors had a broad molecular range, while others were activated only by octanal. Comparison of the profiles with that of the one identified octanal receptor, OR-I7, indicated several differences. While OR-I7 is activated by low concentrations of octanal and blocked by citral, other receptors were less sensitive to octanal and not blocked by citral. A lower estimate for the maximal number of octanal receptors is between 33 and 55. This large number of receptors for octanal suggests that, although the peripheral olfactory system is endowed with high sensitivity, discrimination among different compounds probably requires further central processing.

Acyclic Monoterpenes↗

Noninvasive measurement of steady and pulsating velocity profiles and shear rates in arteries using echo PIV: in vitro validation studies.

Although accurate measurement of velocity profiles, multiple velocity vectors, and shear stress in arteries is important, there is still no easy method to obtain such information in vivo. We report on the utility of combining ultrasound contrast imaging with particle image velocimetry (PIV) for noninvasive measurement of velocity vectors. This method (echo PIV) takes advantage of the strong backscatter characteristics of small gas-filled microbubbles (contrast) seeded into the flow. The method was tested in vitro. The steady flow analytical solution and optical PIV measurements (for pulsatile flow) were used for comparison. When compared to the analytical solution, both echo PIV and optical PIV resolved the steady velocity profile well. Error in shear rate as measured by echo PIV (8%) was comparable to the error of optical PIV (6.5%). In pulsatile flow, echo PIV velocity profiles agreed well with optical PIV profiles. Echo PIV followed the general profile of pulsatile shear stress across the artery but underestimated wall shear at certain time points. However, error in shear from echo PIV was an order of magnitude less than error from current shear measurement methods. These studies indicate that echo PIV is a promising technique for noninvasive measurement of velocity profiles and shear stress.

Animals↗

[Enterohemorrhagic Escherichia coli O157 outbreak in Obihiro-City--study on antibiotic susceptivity and plasmid profiles].

The drug-resistance patterns and plasmid profiles of 147 isolates (patient origin 142 and food origin 5 isolates) from the outbreak of enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection in Obihiro-city Hokkaido in late October, 1996, were examined. Thirty-six isolates were resistant to tetracycline (TC) (24.5%), 15 of which were resistant to both streptomycin and TC. The minimal growth inhibitory concentration (MIC) of fosfomycine (FOM) was examined, confirming that MIC changed by the cultivation conditions, that is 12.5 micrograms/ml at the aerobic condition, 1.6 micrograms/ml at the anaerobic condition and 3.2 micrograms/ml on blood agar plates. Furthermore, though E. coli O157 could not be detected once by the FOM medication, FOM sensitivity of the patient origin O157 isolates who became O157-positive again was examined. Any changes in FOM sensitivity were not observed. Plasmid profiles of all isolates were divided by 4 patterns from A to D. The most dominant pattern was type A, and plasmid profiles of food origin O157 belonged to pattern A. In 9 examples of the person-to-person infection in the family, plasmid patterns of O157 isolates were the same to each other, even though drug-resistant patterns were different. In 13 patients developing the duration of excretion of EHEC, the changes of the drug-resistance patterns were correlated with the changes of plasmid profiles. By comparing plasmid profiles and TC resistance, it was suggested that TC resistance was controlled on a plasmid. Since food origin O157 isolates were sensitive to all drugs and presenting the same plasmid profiles, demonstrating that TC resistance and plasmid are newly added to the bacterial cells while food origin O157 isolates passe inside the human body.

Adolescent↗

A Monte Carlo investigation of fluence profiles collimated by an electron specific MLC during beam delivery for modulated electron radiation therapy.

Modulated electron radiation therapy (MERT) is able to deliver conformal dose to shallow tumors while significantly reducing dose to distal structures and surrounding tissues. An electron specific multileaf collimator (eMLC) has been proposed and constructed as an effective means of delivering electron beams for MERT. The aim of this work is to apply the Monte Carlo method to investigate the fluence profiles collimated by the eMLC in order to achieve accurate beam delivery for MERT. In this work, the EGS4/BEAM code was used to simulate the eMLC collimated electron beams of 6-20 MeV generated from a Varian Clinac 2100C linear accelerator. An attempt was made to describe the fluence profiles with an analytic Sigmoid function. The function parameters were determined by the fittings of the Monte Carlo simulated fluence profiles. How the function parameters depend on the eMLC aperture size, the off-axis location, and the electron beam energy has been investigated. It has been found that the eMLC collimated fluence profiles are dependent on beam energy, while almost independent of leaf location or dimension of MLC aperture. There is little difference in the fluence profiles collimated by the leaf side and the leaf end for the straight-edged leaves. It is possible that these energy-dependent Sigmoid functions can serve as operators to account for the energy dependence of the eMLC collimated fluence profiles. These operators can be incorporated into the inverse planning algorithm to derive desired dose distributions using a set of electron beams of variable energy and field size suitable for delivery by the eMLC.

Computer Simulation↗

Enlarged longitudinal dose profiles in cone-beam CT and the need for modified dosimetry.

In order to examine phantom length necessary to assess radiation dose delivered to patients in cone-beam CT with an enlarged beamwidth, we measured dose profiles in cylindrical phantoms of sufficient length using a prototype 256-slice CT-scanner developed at our institute. Dose profiles parallel to the rotation axis were measured at the central and peripheral positions in PMMA (polymethylmethacrylate) phantoms of 160 or 320 mm diameter and 900 mm length. For practical application, we joined unit cylinders (150 mm long) together to provide phantoms of 900 mm length. Dose profiles were measured with a pin photodiode sensor having a sensitive region of approximately 2.8 x 2.8 mm2 and 2.7 mm thickness. Beamwidths of the scanner were varied from 20 to 138 mm. Dose profile integrals (DPI) were calculated using the measured dose profiles for various beamwidths and integration ranges. For the body phantom (320-mm-diam phantom), 76% of the DPI was represented for a 20 mm beamwidth and 60% was represented for a 138 mm beamwidth if dose profiles were integrated over a 100 mm range, while more than 90% of the DPI was represented for beamwidths between 20 and 138 mm if integration was carried out over a 300 mm range. The phantom length and integration range for dosimetry of cone-beam CT needed to be more than 300 mm to represent more than 90% of the DPI for the body phantom with the beamwidth of more than 20 mm. Although we reached this conclusion using the prototype 256-slice CT-scanner, it may be applied to other multislice CT-scanners as well.

Head↗

A new description of the photon beam peak-depth profile as a function of field size.

The dose profile at peak depth in water is described as the product of an apparatus function and a source function. In principle, the source function is the circularly symmetric profile which would be measured at peak depth without any collimation. In practice, the peak-depth profile in the diagonal plane, measured for the largest collimator setting, is used for this purpose. The apparatus function represents the collimator acting upon the source function, and is referred to as the collimator function. The collimator function for any field size can be developed from the ratio of the peak-depth profile for a single medium-sized field and the source function. The method has been tested for a set of irregularly flattened 4-MV x-ray beams as well as for practically flat 15-MV x-ray beams. The model requires as basic data only three peak-depth profiles: one in each principal plane of a medium-sized square field and the peak-depth profile in the diagonal plane for the largest field. It replaces the peak-depth transformation in the projective beam model.

Humans↗

The measurement of radiation dose profiles for electron-beam computed tomography using film dosimetry.

The unique geometry of electron-beam CT (EBCT) scanners produces radiation dose profiles with widths which can be considerably different from the corresponding nominal scan width. Additionally, EBCT scanners produce both complex (multiple-slice) and narrow (3 mm) radiation profiles. This work describes the measurement of the axial dose distribution from EBCT within a scattering phantom using film dosimetry methods, which offer increased convenience and spatial resolution compared to thermoluminescent dosimetry (TLD) techniques. Therapy localization film was cut into 8 x 220 mm strips and placed within specially constructed light-tight holders for placement within the cavities of a CT Dose Index (CTDI) phantom. The film was calibrated using a conventional overhead x-ray tube with spectral characteristics matched to the EBCT scanner (130 kVp, 10 mm A1 HVL). The films were digitized at five samples per mm and calibrated dose profiles plotted as a function of z-axis position. Errors due to angle-of-incidence and beam hardening were estimated to be less than 5% and 10%, respectively. The integral exposure under film dose profiles agreed with ion-chamber measurements to within 15%. Exposures measured along the radiation profile differed from TLD measurements by an average of 5%. The film technique provided acceptable accuracy and convenience in comparison to conventional TLD methods, and allowed high spatial-resolution measurement of EBCT radiation dose profiles.

Electrons↗

Beam profiles along the nonwedged direction for large wedged fields.

Beam profiles along the nonwedged direction of a wedged field produced by a linear accelerator exhibit more "sagging" than that of an open field at the same depth. For large fields, the profiles of open and wedged fields can differ by as much as 7%. The extra "sagging" of wedged profiles is mainly due to the difference in penetration between on- and off-axis rays caused by the variation of beam quality across the field. An algorithm was developed to estimate an "effective" depth such that the profile of a wedged field can be approximated by the open-field profile at the effective depth. The algorithm was verified by measured beam profiles for 6- and 15-MV x-ray beams for 15 degree, 30 degree, 45 degree, and 60 degree wedges.

Algorithms↗

Electron dose profile shaping by modulation of a scanning elementary beam.

The use of multiple high energy electron beams has been limited in the treatment of deep seated tumors. This is principally because of the rapid increase in the physical electron beam penumbra as a result of the rise in large angle scattering with depth in the patient. This decreases the transverse dose gradient between the target volume and sensitive dose limiting structures and diminishes the ability to conform electron isodose lines to the target volume. If the beam is flat in air, then its profile will become progressively more rounded with depth, due to the increase of scatter out of the beam edges. With a scanning elementary electron beam, such as produced by the Microtron MM50, the characteristics of a broad beam profile are determined by the scan pattern. Using an appropriate scan pattern one can create, at any depth within the range of electrons, various dose profiles with the sharpness not exceeding that of the elementary beam. The objective of this work was to study methods that produce the desired electron beam profile at the depth of the target volume, and to derive the surface fluence profile required. Two approaches were explored to modulate the elementary beam distribution: "amplitude modulation" (AM) and "frequency modulation" (FM). We calculated coordinate and intensity distributions of the 25, 40, and 50 MeV elementary beam pulses at the surface that would yield a flat field at various prescribed depths. The results are in good qualitative agreement with iterative deconvolution calculations by Brahme et al. [Acta Radiol. Oncol. 19, 305-319 (1980)]. The scattering penumbra between the 50%-90% isodose lines can be reduced by up to 40% by beam modulation. The modulation should also enable the combination of multiple electron beams so as to achieve the desired conformal isodose profile as is customarily seen with photon beam planning, but with greater normal tissue sparing due to higher electron longitudinal depth--dose gradients. The results can be also used for electron accelerators that do not use a scanning elementary beam.

Algorithms↗