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Biomedical subjects

M Piechocki

Publications and source records attributed to M Piechocki.

17 recordsLinked to original sources

Progression of head and neck cancer in an in vitro model.

OBJECTIVE: To identify alterations in angiogenesis and cell cycle regulation as preneoplastic cells progress to cancer in an in vitro model of head and neck tumor progression. METHODS: Immortal human gingival keratinocyte (IHGK) cells (preneoplastic) were derived from normal oral keratinocytes and were immortalized with human papillomavirus 16. Transformation of IHGK cells with a carcinogen (NNK, 4-[methylnitrosamino]-1-[3-pyridyl]-1-butanone) gave rise to IHGKN cells. We determined the growth rates, cell cycle phase, expression of cell cycle regulators, and expression of vascular endothelial growth factor along with the organotypic features of these cells and compared them with characteristics of head and neck cancer cells. RESULTS: IHGK and IHGKN cells grown in raft culture were morphologically similar to severe dysplasia and carcinoma, respectively. The proportion of cells in G(0)/G(1) was similar between IHGK and IHGKN. However, the proportion of IHGK cells was 35% greater in S phase as compared with the IHGKN cells, while a greater percentage (40%) of IHGKN cells were in G(2)/M. The expression of the other cell cycle regulators tested was unchanged. IHGK cells secreted less vascular endothelial growth factor on day 1 when compared with IHGKN (50.6 vs 245.6 pg/mL), along with a lower overall production rate (79% vs 133%). CONCLUSIONS: Transformation of IHGK cells resulted in the activation of vascular endothelial growth factor associated with angiogenesis. Inactivation of the G(1) cell cycle regulation occurred during immortalization and before transformation, and was sustained after carcinogen exposure. These alterations correspond to changes observed in patients with head and neck squamous cell carcinoma. This model can be useful in testing novel therapeutic and preventive strategies.

Blotting, Western↗

Localization of basement membrane-associated protein isoforms during development of the ocular surface of mouse eye.

The developmental localization patterns of collagen type IV alpha1-5 chains, laminin-1, laminin-5, and laminin alpha2 chain were analyzed in the embryonic mouse eye using isoform specific antibodies and immunofluorescence microscopy. Laminin-1 isoform and alpha1-2(IV) were ubiquitously expressed along the ocular surface basement membranes at a very early stage of eye development. Alpha3-5(IV) were first detected at later stages of development, and exhibited a variable distribution pattern along the ocular surface basement membrane. In contrast, expression of the laminin alpha2 chain was restricted to the conjunctival basement membrane, and was first detected during the same developmental period in which keratin K4-positive, differentiated conjunctival epithelial cells were observed. Although laminin-5 was uniformly expressed along the adult ocular surface basement membrane, during embryogenesis it was first incorporated into the conjunctival basement membrane structure. These data suggest that some of the laminin isoforms, including laminin alpha2 and laminin-5, may play a role in the formation of a conjunctival-type basement membrane. The temporal relationship between the localization of these molecules to the conjunctival basement membrane and the appearance of differentiated conjunctival epithelial cells suggests a role for external influence on the differentiation pathways of ocular surface epithelium.

Animals↗

The isolation of a yeast artificial chromosome (YAC) contig extending for 2 megabases in the vicinity of the von Hippel Lindau disease gene.

Von Hippel Lindau disease (VHL) is a rare autosomal dominant disease associated with tumors and cysts in multiple organ systems. The VHL disease gene is tightly linked to the polymorphic DNA marker 233E2 (D3S720) and flanked by 479H4 (D3S719) on its telomeric and RAF1 on its centromeric side. Two additional markers, D3S1038 and D3S601, have also been identified, and these markers, like D3S720, are very tightly linked to VHL. Previously 93 cosmid clones were mapped to the larger region, 3p24.2-pter, surrounding the VHL disease gene. Using a Southern-based screening strategy on pools of YAC clones we have isolated a contig of overlapping YAC clones that extends about 0.7 megabase centromeric, and about 1.3 megabases telomeric of D3S720 and contains all three tightly linked VHL markers. Individual YACs in this contig were hybridized to grids containing cosmids localized between 3p24.2-pter and to several cosmids localized by fluorescent in situ hybridization (FISH) to 3p25. A total of 28 cosmids were positioned on this contig of overlapping YAC clones. We have also identified homologous YAC clones to many additional cosmid clones localized between 3p24.2-p25, although these have not yet been precisely localized relative to the contig of YAC clones. This contig of YAC clones probably contains the VHL disease gene and should facilitate the isolation and characterization of this gene.

Base Sequence↗

An attempt to reconstruct the lithotriptor shock wave pulse in kidney: possible temperature effects.

Based on measurements carried out in water in two lithotriptor systems, the authors have made an attempt to reconstruct numerically amplitudes and shapes of shock wave pulses penetrating into kidney which differ from those in water. The difference between these pulses and those observed in water was analyzed and was also demonstrated experimentally. The amplitude and the steepness of the reconstructed pulse front were shown to be much lower than in water depending on the distance of the kidney stone from the patient's body surface. For a distance equal to 4 cm, the shock wave pulse amplitude of 40 MPa in water was estimated to decrease in the kidney by a factor of about two and the steepness of the positive shock pulse front to decrease several times. The analysis was carried out by considering the possible changes of absorption and attenuation in tissues which increase in an unknown way with the wave amplitude. It was shown that the temperature elevation caused by the increase of nonlinear high amplitude absorption is limited due to a corresponding increase in attenuation of the shock wave penetrating soft tissues. The temperature elevation was estimated on the basis of this work to be at most 1.8 times that one estimated in the case of two considered lithotripsy systems when assuming small amplitude absorption and attenuation coefficients.

Animals↗

Smoothed power spectrum estimate applied for analysis of the Doppler signal from blood flow.

A simple method for the improvement of the definition of the instantaneous spectrum estimate of Doppler signal is proposed. A short review of the stochastical properties of FFT spectrum estimates is presented. This review allowed us to develop a concept of the 'estimation noise' as an interpretation of the stochastic uncertainty of the estimation. This, in turn, permitted us to propose a method of adaptive filtering of spectral estimation to minimise the effects of the 'estimation noise'. Proposed filtering in the frequency domain corresponds to a procedure known as smoothing of the estimate. Two different smoothing procedures are presented: classical, linear smoothing and nonlinear, homomorphic smoothing. The performances of the smoothed spectrum estimate are theoretically and experimentally studied, showing that their effectiveness depends mostly on the shape of the Doppler spectrum. Although smoothing always reduces the spectral resolution, the important limitation of the variance of estimation can be achieved without meaningful deterioration of the resolution in our application. Thus, the proposed procedures may sensibly improve the accuracy of the relationship between the shape of the spectrum and the flow parameters. As a result, more exact determination of flow characteristics such as stability or maximum velocity, even in cases of low signal-to-noise power ratio, should be possible.

Algorithms↗

Estimation of the temperature increase in the focus of a lithotripter for the case of high rate administration.

In a recent study Delius et al. demonstrated an increased amount of kidney damage when applying shock waves with a high pulse repetition frequency equal to 100 Hz. The authors interpreted this effect by means in terms of a cavitational mechanism although they could not exclude thermal effects. The present paper estimates the temperature increase for the conditions of the pre-cited study. Taking into account spectral properties of the pressure in the form of cosine, triangle pulses, including their overshoot, as well as an experimental shock wave pulse shape it was possible to compute the temperature increase within the focus of the lithotripter. The maximum estimated temperature increase was equal to 1.8 degrees C for 3000 shock wave pulses of 80 MPa peak pressure. The results of those computations exclude the possibility of significant thermal effects in the damage of kidney tissues even if the administration rate equals 100 shock wave pulses per second.

Animals↗

Estimation of blood-flow quality by statistical analysis or an ultrasonic Doppler signal: application to the study of perturbations caused by a vascular stenosis.

The noninvasive detection of pathological stenoses by Doppler ultrasound velocimetry is based on the appearance of modifications in velocity waveform or of a local increase in velocity. Nevertheless, these methods suffer from a lack of sensitivity. An extension of ultrasonic velocimetry including a statistical treatment of the Doppler signals affords a quantitative approach to the flow quality and seems to be able to improve the diagnosis of vascular obliterans. Accordingly a perturbation index can be computed on a microprocessor as the relative standard deviation of the zero-crossings histogram of the Doppler signal. A theoretical and experimental approach has been attempted to validate this method. Moreover, this index has been tested in vitro on calibrated flows. The in vivo experiments, performed on the abdominal aorta of the dog with artificial stenoses (0 to 50% in diameter) show a significant increase in the index value downstream from the stenosis. The relative increase of the index is greater than that of velocities for the same degree of obstruction. At the moment, it is possible to detect stenoses of 20% and above. It should be noted that changes in the perturbation index can be observed on a large part of the arterial segment, in relation with the severity of teh stenosis. Taking into account the increase in the index value and the length of the disturbed zone downstream of the stenosis, an estimation of the severity itself can be attempted. Clinical applications are in progress pointing out the diagnostic and prognostic abilities of the index method. The index perturbation method adds to velocity measurement the possibility of blood flow stability estimation. It appears useful for the localisation of stenoses, offers the possibility of quantifying their severity and could help the prognosis of their development.

Animals↗

Method for the measurement of the maximum Doppler frequency.

This paper describes a method for detection of the maximum Doppler frequency from a histogram of Doppler signals (HDS). The performance of the system is described, and its behavior in analysis of noise signals that have known lower and upper cut off frequencies is discussed. Experimental investigations show that the device correctly measures the maximum frequency envelope and can be useful in evaluating the maximum flow velocities in the blood circulation system.

Humans↗