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PubMed1992

Frequent deletions in nine newly immortal human cell lines.

Nine newly immortal lines of human fibroblasts transfected with SV40 T antigen were examined for recurrent chromosome losses. In order of decreasing frequency, all nine lines had three or more of the following minimal deletions specifically associated with the immortalization event: del(6)(q21), del(3)(p24), del(1)(p34), del(4)(p25), del(5)(p14), del(11)(p11), del(11)(q14), del(12)(p12), and del(14)(p?). Many other chromosome changes were not clearly associated with immortalization, but were acquired during other stages of this multistep model of neoplastic transformation. We propose that these chromosome loci associated with immortalization are candidates for the location of genes involved in cellular senescence.

Antigens, Polyomavirus Transforming↗
PubMed1996

Computational viewpoint of the OpenLabs architecture.

The computational viewpoint of the OpenLabs architecture specification concentrates on the definition of the functional interfaces between the logical modules that make up the clinical laboratory as defined in the enterprise and information viewpoints. A method is presented which transforms the information models into a specification written in Abstract Syntax Notation One (ASN.1). It is then shown how the specification can be translated into an Interface Definition Language (IDL) of a 'middleware' product which can then be compiled for implementation on a distributed system.

Clinical Laboratory Information Systems↗
PubMed1985

Health care in Costa Rica: boom and crisis.

In 1960-1980 Costa Rica experienced a health boom, achieving significant improvements which moved that country into the number two position in Latin America for indicators such as population coverage, infant mortality, life expectancy and health services. In addition, there was a gradual process of integration of health services. But in the same period, the cost of health care as a percentage of GNP increased almost 5-fold and in 1980 was the fourth highest in the region. The economic crisis of the 1980s aggravated the financial difficulties; to cope with them, the government introduced an austere program to reduce costs and plans to transform the current model of health care into a more efficient one capable of maintaining Costa Rica's high health standards in the future. The paper is divided into five sections: summary of the historical development of health care, and description of its current organization and of its gradual process of integration; estimation of population coverage and its trends, evaluation of inequalities in coverage, and identification of the non-covered group; analysis of health-care financing and its sources, as well of the recent financial desequilibrium, its causes and measures to restore the equilibrium; description of health care benefits and their differences among groups and regions, analysis of the country's advances in health-care facilities and standards, and measurement of the impact of the health care system in income distribution; and description of the rising cost of health care and the current crisis, analysis of the causes of both phenomena, and review of the measures that have been and should be implemented to solve these problems.

Cost Control↗
PubMed2005

The mechanisms of regulation of Hdm2 protein level by serum growth factors.

Cell cycle progression in response to serum growth factors is dependent on the expression of functional Hdm2 (Mdm2), which inhibits p53-dependent transcription of anti-proliferative genes. In a well characterised non-transformed human fibroblast model, growth factors induce the expression of Hdm2 with rapid kinetics. Here we dissect the mechanistic basis for this critical response. In contrast to previous studies in which components of the growth factor signalling pathways were overexpressed, hdm2 mRNA expression is not induced with immediate-early kinetics in these cells. Rather, the elevated Hdm2 protein levels which follow growth factor stimulation are primarily a consequence of phosphatidylinositol-3 kinase-dependent stabilisation of the Hdm2 protein combined with a global increase in protein synthesis.

Cell Cycle↗
PubMed2025

The role of iPSC research for insight into inherited arrhythmia conditions.

Human induced pluripotent stem cells (iPSCs) have emerged as a transformative platform for modeling inherited cardiac arrhythmia syndromes and uncovering human-specific disease mechanisms. However, the promise of iPSC-derived cardiomyocytes lies beyond the recapitulation of arrhythmogenic phenotypes and channelopathies. In this review, we explore recent works which have enabled mechanistic interrogation and therapeutic insight for inherited arrhythmia syndromes, beyond the capabilities of traditional animal models. Such studies have leveraged iPSCs to elucidate the role of splice variants, transcriptional regulation, and mitochondrial stress in arrhythmogenesis. Further, iPSC systems have proven important for reclassifying variants of uncertain significance and in modeling idiopathic arrhythmias where genotype-phenotype links are elusive. Advances in directed differentiation now permit chamber-specific cardiac cell generation, allowing for atrial and ventricular disease modeling and revealing critical cell-cell interactions. iPSCs also serve as high-fidelity precursor platforms for drug testing, offering predictive insight into mutation-specific responses to pharmacologic and genetic therapies. Though limitations in maturation and scalability persist, ongoing efforts for integration with tissue engineering, multi-cellular models, and computational frameworks are evolving to improve model reliability. iPSC-based systems now occupy a critical role in arrhythmia research, bridging basic discovery with translational applications, thereby contributing to personalizing care and advancing therapeutics in inherited and idiopathic arrhythmic syndromes.

Humans↗
PubMed2006

Individually preformed titanium mesh implants for a true-to-original repair of orbital fractures.

The purpose of this investigation is to present the results using preoperatively-formed titanium mesh implants for a true-to-original primary repair of extensive orbital floor and medial wall fractures. Individually preformed implants were used to repair extensive orbital floor injuries in 19 patients at the University Hospital, Freiburg. The form of the orbital floor and walls was analysed by preoperative diagnostic CT scan data. The form of the virtual reconstructed orbit was transformed into a model of the orbital cavity by a template machine. Postoperative imaging by or CT scan verified the exact 3D reconstruction of the orbital cavity 'true to original'. None of the patients demonstrated diplopia or enophthalmos postoperatively. Using individually preformed titanium mesh implants, the accuracy of the 3D orbital reconstruction was within a range of 1mm. The reconstruction using preformed implants proved to be less time consuming, more precise and less invasive, compared to 'free hand' efforts, for the repair of orbital injuries using titanium mesh and calvarial grafts.

Biocompatible Materials↗
PubMed2006

Clinical experience with a 3D surface patient setup system for alignment of partial-breast irradiation patients.

PURPOSE: To assess the utility of surface imaging on patient setup for accelerated partial-breast irradiation (APBI). METHODS AND MATERIAL: A photogrammetry system was used in parallel to APBI setup by laser and portal imaging. Surface data were acquired after laser and port-film setup for 9 patients. Surfaces were analyzed in comparison to a reference surface from the first treatment session by use of rigid transformations. The surface model after laser setup was used in a simulated photogrammetry setup procedure. In addition, breathing data were acquired by surface acquisition at a frame rate of 7 Hz. RESULTS: Mean 3D displacement was 7.3 mm (SD, 4.4 mm) and 7.6 mm (SD, 4.2 mm) for laser and port film, respectively. Simulated setup with the photogrammetry system yielded mean displacement of 1 mm (SD, 1.2 mm). Distance analysis resulted in mean distances of 3.7 mm (SD, 4.9 mm), 4.3 mm (SD, 5.6 mm), and 1.6 mm (SD, 2.4 mm) for laser, port film, and photogrammetry, respectively. Breathing motion at isocenter was smaller than 3.7 mm, with a mean of 1.9 mm (SD, 1.1 mm). CONCLUSIONS: Surface imaging for PBI setup appears promising. Alignment of the 3D breast surface achieved by stereo-photogrammetry shows greater breast topology congruence than when patients are set up by laser or portal imaging. A correlation of breast surface and CTV must be quantitatively established.

Breast Neoplasms↗
PubMed2006

Expression of a novel gene, MafB, in Dupuytren's disease.

PURPOSE: Dupuytren's disease (DD) is characterized by fibroblastic proliferation of the palmar fascia, often leading to flexion contracture in the hand. Although there is a strong genetic component the genome-wide expression of novel genes is not known. The purpose of this study was to use DNA microarray technology to identify upregulated genes in DD. METHODS: Human tissue samples were harvested from 3 patient sources: DD cord tissue (n = 20), normal-appearing adjacent control fascia (n = 15), and palmar fascia from patients having carpal tunnel release (n = 15). DNA microarray analysis was performed on amplified sample RNA. Novel genes were compared with known gene functions. A candidate gene of interest was studied further by using immunohistochemistry on DD tissue samples and controls. RESULTS: Several novel genes not described previously in the study of DD were upregulated significantly, including MafB, collagen type V, alpha-2 (COL5A2), collagen type VIII, alpha-1 (COL8A1), contactin I (CNTN1), and leucine-rich repeat containing 17 (LRRC17). These upregulated genes were compared with their known gene-expression profiles in other tissues and their purported functions. MafB was found to be of particular interest because of its prominent role in tissue development and cellular differentiation. MafB immunohistochemistry showed positive staining in 50% of the DD specimens but complete absence of MafB in all control tissues (adjacent control fascia, carpal tunnel fascia). Co-localization experiments with MafB and alpha-smooth muscle actin showed staining properties in similar regions but these 2 proteins were not confined solely to the same cells. CONCLUSIONS: Microarray analysis of DD tissue has identified significant upregulated gene expression of MafB. MafB protein also is found in Dupuytren's cords but not in control fascia. Co-localization data suggest that the association of MafB with DD is not related exclusively to myofibroblast proliferation. Because of its role in fibroblastic transformation in other models MafB and its relationship to the pathogenesis of DD deserves further study.

Actins↗
PubMed2005

BMP7 signaling in renal development and disease.

Fibrosis, and in particular tubulointerstitial fibrosis, is a common feature of almost all chronic renal diseases. Over the past several years, significant progress has been made in defining the underlying mechanisms of tubulointerstitial fibrosis. In a variety of mouse models, expression of transforming growth factor-beta is a primary causative factor which leads to increased numbers of myofibroblasts, collagen deposition and loss of tubular epithelia. More recently, another member of the transforming growth factor-beta superfamily, BMP7, was shown to counteract transforming growth factor-beta-mediated fibrosis. The activities of these secreted factors are regulated, in part, by extracellular ligand binding proteins which can enhance or suppress receptor ligand interactions.

Animals↗
PubMed2004

Influence of MRI acquisition protocols and image intensity normalization methods on texture classification.

Texture analysis methods quantify the spatial variations in gray level values within an image and thus can provide useful information on the structures observed. However, they are sensitive to acquisition conditions due to the use of different protocols and to intra- and interscanner variations in the case of MRI. The influence was studied of two protocols and four different conditions of normalization of gray levels on the discrimination power of texture analysis methods applied to soft cheeses. Thirty-two samples of soft cheese were chosen at two different ripening periods (16 young and 16 old samples) in order to obtain two different microscopic structures of the protein gel. Proton density and T(2)-weighted MR images were acquired using a spin echo sequence on a 0.2 T scanner. Gray levels were normalized according to four methods: original gray levels, same maximum for all images, same mean for all images, and dynamics limited to micro +/- 3sigma. Regions of interest were automatically defined, and texture descriptors were then computed for the co-occurrence matrix, run length matrix, gradient matrix, autoregressive model, and wavelet transform. The features with the lowest probability of error and average correlation coefficient were selected and used for classification with 1-nearest neighbor (1-NN) classifier. The best results were obtained when using the limitation of dynamics to micro +/- 3sigma, which enhanced the differences between the two classes. The results demonstrated the influence of the normalization method and of the acquisition protocol on the effectiveness of the classification and also on the parameters selected for classification. These results indicate the need to evaluate sensitivity to MR acquisition protocols and to gray level normalization methods when texture analysis is required.

Cheese↗
PubMed2004

Biocenosis of BAC(F)s used for groundwater treatment.

The studies on the biocenosis of biologically active carbon filters (BAC(F)) used in treatment of Miocene water revealed the presence of protozoa of the group of flagellates and ciliates (Glaucoma sp., Opercularia sp.), saprophytic bacteria, phase I and II nitrifying bacteria, Fe(II) and Mn(II) oxidizing bacteria of the family Siderocapsaceae, Leptothrix ochracea and Pedomicrobium sp., as well as microscopic fungi. The stratification of biofilter colonisation by these microorganisms was found to be similar to that occurring in biofilters applied in sewage treatment. A hypothetical model of microbiological transformations in BAC(F)s, brought about by various physiological groups of microorganisms, is presented. It is shown that ozone pre-treatment of water dosed to the biofilter reduces the number of saprophytic bacteria and moulds in its upper layer, as well as Fe(II) oxidizing bacteria of the family Siderocapsaceae across the full section of the biofilter; it does not, however, influence the abundance of nitrifying phase I and II autotrophic bacteria and Mn(II) oxidizing bacteria of the family Siderocapsaceae. The abundance of microorganisms in the biofilter outflow is increased in comparison with that in untreated water; they do not, however, create a health hazard for human beings.

Animals↗
PubMed2004

Spectral analysis of internal carotid arterial Doppler signals using FFT, AR, MA, and ARMA methods.

In this study, Doppler signals recorded from internal carotid artery of 45 subjects were processed by PC-computer using classical (fast Fourier transform) and model-based (autoregressive, moving average, autoregressive moving average (ARMA) methods) methods. Power spectral density estimates of internal carotid arterial Doppler signals were obtained using these spectral analysis methods. The variations in the shape of the Doppler power spectra as a function of time were presented in the form of sonograms in order to determine the degree of internal carotid artery stenosis. These Doppler power spectra and sonograms were then used to compare the applied methods in terms of their frequency resolution and the impact on determining stenosis in internal carotid arteries. Based on the results, performance characteristics of the autoregressive and ARMA methods were found extremely valuable for spectral analysis of internal carotid arterial Doppler signals obtained from healthy subjects and unhealthy subjects having artery stenosis.

Adolescent↗
PubMed1998

High sample throughput flow immunoassay utilising restricted access columns for the separation of bound and free label.

A flow immunodetection system with high sample throughput capacity is described for the screening of various analytes. The immunochemical detection principle is based on the chromatographic separation of the formed immunocomplex (AbAg or AbAg*) and the free antigen (Ag) by a restricted access (RA) column, utilising size-exclusion and reversed-phase mechanism. A fluorescein labelled analyte (Ag*) was used in the competitive assay format with fluorescence detection. The speed and simplicity of the assay were the greatest advantages, allowing measurement of the analyte to be carried out in less than 1 min. The biocompatibility and capacity of the restricted access material allowed multiple injections of up to 5000, without any breakthrough of the fluorescent tracer molecule and thus need for regeneration. The flow immunoassay was developed using the well-known atrazine herbicide and some transformation products as model compounds, due to their human toxicity and widespread use. The sample throughput was 80 samples per hour and the detection limits were 1.4 nM (300 pg/ml) for atrazine (Ab I) and 2.3 nM (500 pg/ml) for the sum of triazines (Ab II-III). Different sample matrices, PBS buffer, creek water, and urine were successfully applied in the flow system without the need for any sample handling step. For plasma samples an additional clean-up step using solid-phase extraction had to be included. The resulting detection limits for atrazine in plasma and water samples using this clean-up and trace enrichment procedure were found to be 2 ng/ml and 20 pg/ml, respectively. The analysis could be performed at a sample throughput rate of 400 per 6-h working shift.

Atrazine↗
PubMed2002

Neuropsychological perspectives on eye-hand coordination in visually-guided reaching.

Substantial progress has been made in understanding the neural control of movement in the past 30 years. Lower cost technology for tracking movements of the eyes and the hands has increased our understanding of these two systems and their interactions in both neurologically intact individuals and non-human primates. Nevertheless the neuropsychology of eye-hand coordination during visually-guided tasks such as reaching and grasping remains relatively understudied. This chapter reviews some of the relevant neurophysiology and neuropsychology of eye-hand coordination during visually-guided reaching. Current models emphasising coordinate transformations are discussed in light of new patient data showing a particular type of failure of eye-hand coordination during reaching.

Animals↗
PubMed1998

Human T cell leukemia virus type 1 oncoprotein Tax targets the human mitotic checkpoint protein MAD1.

In searching for cellular targets of the HTLV-I oncoprotein Tax, we identified TXBP181, which we characterized as the human homolog of yeast mitotic checkpoint MAD1 protein. Evidence supporting TXBP181 as HsMAD1 includes sequence conservation with yeast MAD1, hyperphosphorylation during S/G2/M phases and upon treatment of cells with nocodazole, and binding to HsMAD2. HsMAD1 functions as a homodimer. It localizes to the centrosome during metaphase and to the spindle midzone and the midbody during anaphase and telophase. Expression of either Tax or a transdominant-negative TXBP181 results in multinucleated cells, a phenotype consistent with a loss of HsMAD1 function. We propose a model of viral transformation in which Tax targets TXBP181, thereby abrogating a mitotic checkpoint.

Amino Acid Sequence↗
PubMed2003

Involvement of protein phosphatase 2A in interferon-alpha-2b-induced apoptosis in K562 human chronic myelogenous leukaemia cells.

Interferon-alpha (IFN-alpha)-2b is known to have antiproliferative effects on hematological malignant cells, especially chronic myelogenous leukaemia (CML). However, it can induce cytogenetical remissions in a very small percentage of the patients. Also during interferon therapy, resistance can emerge in the CML clones. K562 is an in vitro model cell line transformed from a Ph positive CML patient. It can be induced to differentiate to granulocytic and/or monocytic lineages with certain molecules. IFN-alpha-2b generally exerts its effects on CML cells by Janus family kinases (Jak/Stat) pathway, mostly through tyrosine kinase system. However, there is almost no data on the relevance of serine/threonine (Ser/Thr) protein phosphatase (PP) system in the interferon induced signal transduction pathways. In this study, we investigated serine/threonine protein phosphatases in the IFN-alpha-2b induced K562 cytotoxicity. Trypan blue dye exclusion test and MTT assay were utilised for determining cytotoxicity. IC(50) of IFN-alpha-2b on K562 cells was found to be 600IU/ml. However, no differentiation was determined by analysis of cell surface antigen expressions. Serine/threonine protein phosphatase inhibitors calyculin A (Cal A) and okadaic acid (OKA) augmented the IFN-alpha-2b induced cytotoxicity. Apoptosis assay by the mono-oligonucleosome detection and acridine orange/propidium iodide dye revealed marked apoptosis underlying cytotoxicity. Phosphatase enzyme assay revealed a gradual increase in protein phosphatase 2A (PP2A) activity during interferon induced cytotoxicity. Conversely, immunoblots showed no change in the expression of PP2A catalytic and regulatory subunits. In conclusion, PP2A plays a role in IFN-alpha-2b induced apoptosis of K562 cells and should be investigated as a new window furthermore.

Apoptosis↗
PubMed2001

Influence of dispersants on the bioavailability and trophic transfer of petroleum hydrocarbons to larval topsmelt (Atherinops affinis).

Use of chemical dispersants as oil spill clean-up agents may alter the normal behavior of petroleum hydrocarbons (PH) by increasing their functional water solubility, resulting in increased bioavailability and altered interactions between dispersant, oil, and biological membranes. The objective of this research was to determine the impact of dispersing agents on PH bioavailability and trophic transfer to larval fish from primary levels of a marine food chain. Uptake, bioaccumulation, depuration, and metabolic transformation of a model PH, [14C]naphthalene, were measured and compared for Prudhoe Bay crude oil (PBCO) dispersed with Corexit 9527(R) (DO) and undispersed preparations of the water-accommodated fraction (WAF) of PBCO. The model food chain consisted of a primary producer, Isochrysis galbana; and a primary consumer, the rotifer, Brachionus plicatilis; and larval topsmelt, Atherinops affinis. Direct aqueous (AQ) exposure was compared with combined aqueous and dietary (AQ&D) exposure. Dispersants altered the uptake and depuration processes of naphthalene, independent of aqueous concentrations, in primary trophic species of a marine food chain. The amount of naphthalene taken up by topsmelt was initially significantly (P < or = 0.05) enhanced in the presence of dispersant, reaching a maximum more quickly than undispersed samples. Dispersion treatment significantly increased naphthalene dispension in topsmelt (P < or = 0.05) from both AQ and AQ&D exposures. No significant change in naphthalene uptake by fish was observed with the addition of contaminated food for either WAF or DO medium; however, both uptake and depuration rate constants varied significantly with route of exposure consistent with greater naphthalene turnover. The majority (> or = 72%) of naphthalene-derived radioactivity from fish tissue following all exposures was in the parent form, with smaller quantities of alpha- and beta-naphthols, alpha- and beta-naphthyl sulfates, and an unidentified derivative.

Animals↗