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At least 217 records · Page 12Linked to original sources
PubMed1989

Continuous maintenance of transformed fibroblasts under reduced serum conditions: utility as a model system for investigating growth factor-specific effects in nonquiescent cells.

We report the continuous growth maintenance of untransformed and chemically transformed fibroblasts (AKR-2B, AKR-MCA cells) in low concentrations of serum (0.1% FBS). The cell lines established (AKR-0.1F, MCA-0.1F) proliferated at rates comparable to cells maintained under high serum conditions (10% FBS). Complete removal of serum from the cells did not induce quiescence. The MCA-0.1F cells were more similar to the untransformed AKR-2B fibroblasts in their morphology, saturation density, inability to form colonies under anchorage-independent conditions, steady-state level of c-myc expression, and kinetics of induction of c-myc in response to specific growth factors. This report demonstrates the utility of this cell line as a nonquiescent model system for investigating growth factor-specific effects in serum-free, cycling cells. Addition of transforming growth factor-beta (TGF-beta) (5 ng/ml) to proliferating MCA-0.1F cells, in the absence of any serum, induced a multilayered growth pattern at confluency, similar to that of AKR-MCA cells maintained in 10% FBS. Other growth factors tested did not elicit this effect. The induction of this growth pattern by TGF-beta was associated with a sustained induction of the c-myc proto-oncogene at confluency, but not with a restoration of anchorage-independent growth. The data suggest that TGF-beta may play a role in the up-regulation of c-myc at confluency previously described for AKR-MCA cells maintained in 10% serum.

Animals↗
PubMed1992

Transformation of a continuous rat embryo fibroblast cell line requires three separate domains of simian virus 40 large T antigen.

Mouse C3H 10T1/2 cells and the established rat embryo fibroblast cell line REF-52 are two cell lines widely used in studies of viral transformation. Studies have shown that transformation of 10T1/2 cells requires only the amino-terminal 121 amino acids of simian virus 40 (SV40) large T antigen, while transformation of REF-52 cells requires considerably more of large T antigen, extending from near the N terminus to beyond residue 600. The ability of a large set of linker insertion, small deletion, and point mutants of SV40 T antigen to transform these two cell lines and to bind p105Rb was determined. Transformation of 10T1/2 cells was greatly reduced by mutations within the first exon of the gene for large T antigen but was only modestly affected by mutations affecting the p105Rb binding site or the p53 binding region. All mutants defective for transformation of 10T1/2 cells were also defective for transformation of REF-52 cells. In addition, mutants whose T antigens had alterations in the Rb binding site showed a substantial reduction in transformation of REF-52 cells, and the degree of this reduction could be correlated with the ability of the mutant T antigens to bind p105Rb. There was a tight correlation between the ability of mutants to transform REF-52 cells and the ability of their T antigens to bind p53. These results demonstrate that multiple regions of large T antigen are required for full transformation by SV40.

Animals↗
PubMed2004

Transformation diagrams for the collapse of a phospholipid monolayer.

The kinetics of phase transitions in three-dimensional bulk materials are commonly presented in transformation diagrams. Time-temperature transformation (TTT) and continuous-cooling-transformation (CCT) diagrams plot the time required to transform specific fractions of the material to the new phase by cooling below a transition temperature. Transformation occurs isothermally for the TTT diagrams and during continuous cooling through a range of temperatures for CCT curves. Here we present analogous transformation diagrams for two-dimensional monolayers, which collapse at the equilibrium spreading pressure (pi e) to form a three-dimensional bulk phase. Time-surface pressure-transformation (TpiT) diagrams give the time required for specific fractions of the film to collapse when surface pressure is constant, and continuous-compression-transformation diagrams give the same information when surface pressure varies continuously. The diagrams, constructed here from previously reported data for 1-palmitoyl-2-oleoyl phosphatidylcholine, provide insights into the behavior of the films. The TpiT diagrams successfully predict the existence and approximate magnitude of a threshold rate for compressing the films to high surface pressures above pi e and the approximate shape of isotherms obtained with different rates of interfacial compression. The diagrams also caution that the behavior of mixed monolayers, explained previously in terms of compositional changes, can instead result from collapse that varies with surface pressure. Finally, the similarity between the shapes of the TTT and TpiT diagrams, with the time for transformation passing through a minimum and then increasing as the systems deviate further from equilibrium, suggests that analogous mechanisms determine the behavior of both systems.

In Vitro Techniques↗
PubMed1997

Transformed cells require continuous activity of RNA polymerase II to resist oncogene-induced apoptosis.

Studies have indicated that deregulated oncogene expression can result in either programmed cell death or proliferation, depending on the cellular microenvironment. However, little is known about whether oncogenic signals in themselves are able to activate a cellular apoptotic program. We have tested the hypothesis that oncogenic signals in the absence of gene expression are sufficient to induce cell death, which would indicate that constitutive expression of antiapoptotic genes is necessary for maintenance of the transformed state. Using two highly specific RNA polymerase (RNAP) II inhibitors, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) and alpha-amanitin, which inhibit RNAP II function by two distinct mechanisms, we found that inhibition of gene expression substantially increased apoptosis in a time- and dose-dependent manner in p53+/+- and p53(-/-)-transformed mouse embryonic fibroblasts and in HeLa cells, demonstrating that this type of apoptosis does not require wild-type p53. Engineered expression of an alpha-amanitin resistance RNAP II gene rendered cells resistant to induction of apoptosis by alpha-amanitin without affecting their sensitivity to DRB, indicating that alpha-amanitin induces apoptosis solely by inhibiting RNAP II function and not by a nonspecific mechanism. DRB-induced apoptosis was independent of the cell cycle or ongoing DNA replication, since DRB induced similar levels of apoptosis in asynchronous cells and cells synchronized by collection at mitosis. Inhibition of RNAP II in untransformed cells like Rat-1 or human AG1522 fibroblasts resulted not in apoptosis but in growth arrest. In contrast, deregulated expression of c-Myc in Rat-1 cells dramatically increased their sensitivity to DRB, directly demonstrating that apoptosis following inhibition of RNAP II function is greatly enhanced by oncogenic expression. The requirement for RNAP II function to prevent oncogene-induced apoptosis implies the need for the constitutive expression of an antiapoptotic gene(s) to maintain the transformed state. The differential sensitivities of untransformed and transformed cells to induction of apoptosis by transcriptional inhibition, coupled with the finding that this type of apoptosis is independent of p53 status, suggest that inhibition of RNAP II may be exploited therapeutically for the design of successful antitumor agents.

Amanitins↗
PubMed1999

Molecular alterations of the Fyn-complex occur as late events of human T cell activation.

Two-dimensional gel electrophoresis of anti-p59fyn immunoprecipitates obtained from non-transformed resting human T lymphocytes resulted in the identification of an oligomeric protein complex which is constitutively formed between Fyn and several additional phosphoproteins (pp43, pp72, pp85, the protein tyrosine kinase Pyk2, as well as the two recently cloned adaptor proteins, SKAP55 and SLAP-130). With the exception of pp85, these proteins seem to preferentially interact with Fyn since they are not detectable in Lck immunoprecipitates prepared under the same experimental conditions. Among the individual members of the Fyn-complex pp85, SKAP55 and pp43 are constitutively phosphorylated on tyrosine residue(s) in vivo and likely interact with Fyn via its src homology 2 (SH2)-domain. In contrast to non-transformed T lymphocytes, continuously proliferating transformed human T cell lines express an altered Fyn-complex. Thus, despite normal expression and tyrosine phosphorylation, SKAP55 does not associate with Fyn in Jurkat cells and in other human T cell lines. Instead two novel proteins interact with Fyn among which one has previously been identified as alpha-tubulin. Importantly, almost identical alterations of the Fyn-complex as observed in Jurkat cells are induced in non-transformed T lymphocytes following mitogenic stimulation. These data suggest that Fyn and its associated proteins could be involved in the control of human T cell proliferation. Moreover, the analogous constitutive alterations in transformed T cell lines could indicate that deregulation of the Fyn-complex might be functionally associated with the malignant phenotype of these cells.

Adaptor Proteins, Signal Transducing↗
PubMed2004

RRM analysis of protoporphyrinogen oxidase.

Enzymes are crucial in accelerating metabolic reactions in living organisms. Protoporphyrinogen oxidase (PpOI) is an enzyme that catalyses the production of protoporphyrin IX (PpIX), a protein used in a cancer treatment known as photodynamic therapy (PDT). In this study, a structure-function analysis of PpOI was carried out using the Resonant Recognition Model (RRM), a physico-mathematical approach for analysis of proteins interactions. This method is based on the finding that the distribution of delocalised electron energies along the protein plays a crucial role in determining the protein's biological activity. Two digital signal processing (DSP) methods were used: Fourier Transform (FT) and Continuous Wavelet Transform (CWT). Here we have determined the characteristic frequencies and the "hot spot" amino acids, and predicted the location of proteins' active site(s). Several proteins that potentially belong to the PpOI functional group were also analysed to distinguish their viability in this role.

Algorithms↗
PubMed2004

Reorganizational dynamics of multilamellar lipid bilayer assemblies using continuously scanning Fourier transform infrared spectroscopic imaging.

We employ an implementation of rapid-scan Fourier transform infrared (FT-IR) microspectroscopic imaging to acquire time-resolved images for assessing the non-repetitive reorganizational dynamics of aqueous dispersions of multilamellar lipid vesicles (MLVs) derived from distearoylphosphatidylcholine (DSPC). The spatially and temporally resolved images allow direct and simultaneous determinations of various physical and chemical properties of the MLVs, including the main thermal gel to liquid crystalline phase transition, comparisons of vesicle diffusion rates in both phases and the variation in lipid bilayer packing properties between the inner and outer lamellae defining the vesicle. Specifically, in the lipid liquid crystalline phase, the inner bilayers of the MLVs are more intermolecularly ordered than the outer regions, while the intramolecular acyl chain order/disorder parameters, reflecting the overall characteristics of the fluid phase, remain uniform across the vesicle diameter. In contrast, the lipid vesicle gel phase displays no intermolecular or intramolecular dependence as a function of distance from the MLV center.

Diffusion↗
PubMedDate not supplied

Analysis and study of the variation of splitting in the second heartbeat sound of wavelet transform.

The second heart sound, S2, consists of two acoustic components, A2 and P2. The former is due to the closure of the aortic valve and the latter is due to the closure of the pulmonary valve. The aortic valve usually closes before the pulmonary valve, introducing a time delay known as the 'split'. A technique based on discrete wavelet transform (DWT) and continuous wavelet transform (CWT) is developed in this paper to measure the split. To quantify splitting, two components in S2 (i.e. A2 and P2) are identified, and the delay between the two components can be estimated. One normal case and three pathological cases (mitral stenosis, pulmonary stenosis and atrial septal defect) are considered in this study. The split is measured for each S2 sound of the considered signals. The split normally varies in duration over the cardiac cycle. In certain pathologies such as ASD (atrial septal defect) or PS (pulmonary stenosis), the split becomes fixed over the cardiac cycle. The main part of this paper consists of the identification and measurement of the S2 split. The study confirms the notion of 'variable splitting' for normal phonocardiogram and 'fixed splitting' for ASD and PS cases. This paper relates also to the establishment of statistical parameters to make a distinction between normal and pathological cases of phonocardiogram signals.

Aortic Valve↗
PubMed1994

Unsaturated fatty acids are required for continuous proliferation of transformed androgen-dependent cells by fibroblast growth factor family proteins.

Increase in dietary fat intake has been reported to be associated with progression of hormone-dependent cancers. To explore its mechanism, we examined the effects of fatty acids on the growth of androgen-dependent SC-3 cells cloned from mouse mammary cancer (Shionogi carcinoma 115). Their androgen-dependent growth was potentiated by linoleic acid in the defined medium. The effect of linoleic acid on fibroblast growth factor (FGF)-dependent growth was also addressed because androgen had been demonstrated to exert its mitogenic activity on SC-3 cells through an induction of the unique FGF family protein termed as androgen-induced growth factor. Exposure of SC-3 cells to basic FGF or androgen-induced growth factor exhibited only transient growth response. However, simultaneous addition of linoleic acid to the medium sustained the proliferation of FGF-stimulated, but not FGF-unstimulated, cells, although linoleic acid did not exert the significant effect on the process of S-phase entry of basic FGF-stimulated cells. Palmitoleic acid and oleic acid appeared to exert the actions similar to linoleic acid, while stearic acid was without any effect. Neither cyclooxygenase inhibitor nor 5-lipoxygenase inhibitor could block the growth-promoting ability of linoleic acid. Linoleic acid also enhanced their anchorage-independent growth in the presence of basic FGF. These results indicate that these unsaturated fatty acids play a role in sustaining the proliferation of FGF-stimulated SC-3 cells.

Animals↗
PubMed2000

Wavelet analysis of transient biomedical signals and its application to detection of epileptiform activity in the EEG.

Wavelet based signal analysis provides a powerful new means for the analysis of nonstationary signals such as the human EEG. The properties of the discrete wavelet transform are reviewed in illustrated application examples. The continuous wavelet transform is shown to provide better detection and representation of isolated transients. An approach to extract features of edges and transients from the continuous wavelet transform is outlined. Matching pursuit is presented as a more general transform method that covers both transients and oscillation spindles. A statistical model for the continuous wavelet transform of background EEG is found. A spike detection system based on this background model is presented. The performance of this detection system has been assessed in a preliminary clinical study of 11 EEG recordings containing epileptiform activity and shown to have a sensitivity of 84% and a selectivity of 12%. The spatial context of epileptiform activity will be incorporated to improve system performance.

Brain↗
PubMed2000

Phase transformation behavior in a multipurpose dental casting gold alloy during continuous heating.

Phase transformation in a multipurpose dental casting gold alloy during continuous heating was studied by electrical resistivity measurements, hardness tests, X-ray diffraction and scanning and transmission electron microscopy. The behavior can be explained by the following reaction sequences in the nodule: alpha1(fcc) + alpha2(L1(2)) --> alpha1(fcc) + alpha2(L1(2)) + beta(L1(0)), where fcc is face centred cubic. A discontinuous precipitation with very fine nodules contributed to the hardening and the growth produced the softening. This multipurpose gold alloy is characterized by the introduction of a PtZn ordered phase with L1(0) structure instead of a CuAu I phase.

Journal Article↗
PubMed1989

Continuous-wave ultrasound and neoplastic transformation in vitro.

C3H/10T1/2 cells in suspension were assayed using an initiation-promotion protocol for neoplastic transformation induced by continuous-wave ultrasound. Cells were insonated at 1.765 MHz for 40 min. Two ultrasonic intensities were used: 1.3 and 2.6 W/cm2 spatial average. The first intensity was found to be noncytotoxic; the second resulted in immediate lysis of 20% of the cells, followed by the clonogenic survival of 64% of the remaining cells. Ultrasound was delivered alone or in combination with X-rays (2 Gy, 240 kVp given before ultrasound), and/or 12-O-tetradecanoyl-phorbol-13-acetate (TPA, 0.1 microgram/ml post-irradiation). Under all treatment conditions, there was no effect of ultrasound on transformation at the 95% confidence level.

Animals↗
PubMed2005

Validation and refinement of mortality risk models for heart valve surgery.

BACKGROUND: The Northern New England Cardiovascular Disease Study Group (NNE) recently published risk models for hospital mortality after heart valve surgery. The Providence Health System Cardiovascular Study Group (PHS) has been collecting similar heart valve data for 8 years, providing an ideal opportunity to both validate the NNE risk models and attempt to produce an improved model, by using some different modeling techniques. METHODS: From 1997 to 2004, 3,324 patients aged 30 to 95 years underwent aortic valve replacement (AVR), and 1,596 underwent mitral valve replacement or repair (MVRR) at one of nine PHS medical centers. We used area under the receiver operating characteristic curve (c-index) to measure model discrimination, and Hosmer-Lemeshow statistic (H-L) to measure calibration. We modified the NNE models by ungrouping continuous variables, seeking optimal transformations of continuous variables, and imputing missing values by multiple regression. RESULTS: The prevalence and the lethality of risk factors were similar in PHS and NNE patients. The NNE models fit PHS patients well: c-index (95% confidence interval) = 0.75 (0.70 to 0.80) for AVR and 0.81 (0.76 to 0.86) for MVRR; H-L = 3.95 (p = 0.861) for AVR and 7.10 (p = 0.526) for MVRR. A single PHS model performed slightly better for both positions: c-index = 0.79 (0.75 to 0.83) for AVR and 0.84 (0.80 to 0.88) for MVRR; H-L = 2.75 (p = 0.949) for AVR and 12.21 (p = 0.142) for MVRR. CONCLUSIONS: The NNE models for aortic and mitral valve surgery were successfully validated using PHS patients. Using some different statistical approaches to modeling, we produced a new, unified model for both positions.

Adult↗
PubMed2006

Automatic measure of the split in the second cardiac sound by using the wavelet transform technique.

This paper is concerned with the identification and automatic measure of the split in the second heart sound (S2) of the phonocardiogram signal (PCGs) for normal or pathological case. The second heart sound S2 consists of two acoustic components A2 and P2, the former is due to the closure of the aortic valve and the latter is due to the closure of the pulmonary valve. The aortic valve usually closes before the pulmonary valve, introducing a time delay known as "split". A automatic technique based on the discrete wavelet transform (DWT) and the continuous wavelet transform (CWT) is developed in this paper to measure the split of the second cardiac sound (S2) for the normal and pathological cases of the PCG signals. To quantify the splitting, the two components in S2 (i.e. A2 and P2) are identified and, the delay between the two components can be estimated. It is shown that the wavelet transform can provide best information and features of the split of S2 and the major components (A2 and P2) and consequently aid in medical diagnosis.

Aortic Valve↗
PubMed2006

Determination of carotid disease with the application of STFT and CWT methods.

In this study, Doppler signals were recorded from the output of carotid arteries of 40 subjects and transferred to a personal computer (PC) by using a 16-bit sound card. Doppler difference frequencies were recorded from each of the subjects, and then analyzed by using short-time Fourier transform (STFT) and the continuous wavelet transform (CWT) methods to obtain their sonograms. These sonograms were then used to determine the relationships of applied methods with medical conditions. The sonograms that were obtained by CWT method gave better results for spectral resolution than the STFT method. The sonograms of CWT method offer net envelope and better imaging, so that the measurement of blood flow and brain pressure can be made more accurately. Simultaneously, receiver operating characteristic (ROC) analysis has been conducted for this study and the estimation performance of the spectral resolution for the STFT and CTW has been obtained. The STFT has shown a 80.45% success for the spectral resolution while CTW has shown a 89.90% success.

Carotid Artery Diseases↗
PubMed1977

A continuous line of Rous sarcoma virus-transformed chick embryo cells.

A continuous line of Rous sarcoma virus (RSV)-transformed chick embryo cells was established. The cells, designated RTAZ-1, which constitute the only known line of continuously growing RSV-transformed cells of chick embryo origin, grow rapidly, display uniform morphology, and perpetually release large amounts of RSV (Rous-associated virus, type 1). RTAZ-1 cells display a heteroploid chromosome complement with 92-94 chromosomes characteristic of chicken cells. Molecular hybridization studies demonstrated that DNA from these cells hybridized with normal chick DNA but not with human (KB) cell DNA, thus providing additional evidence of the chicken origin of RTAZ-1.

Animals↗