Tumour-promoting activity of altered WWP1 expression in breast cancer and its utility as a prognostic indicator

WWP1 is a ubiquitin ligase, associated with the post-translational regulation of several tumour-promoting and tumour suppressor proteins. Here we show that WWP1 expression is up-regulated in a subset of breast tumour cell lines and primary breast tumours. We overexpressed WWP1 in MCF10A breast epithelial cells and demonstrated increased cell growth and anchorage-independent colony formation. RNAi knockdown of WWP1 expression in T47D and MCF7 breast tumour cell lines reduced anchorage-independent colony formation. We used WWP1 protein expression levels, in combination with its sub-cellular localization, to classify breast tumours into four categories. Surprisingly, a category with low/absent WWP1 expression displayed a consistently worse prognosis compared with WWP1-expressing tumours. Importantly, the association with disease-free survival was independent of the status of other commonly used prognostic indicators. Thus, WWP1 is a prognostic marker and may be a potential therapeutic target for a subset of breast tumours.     

Optical characterization of a radiochromic film by total reflectance and transmittance measurements

The GafChromic film (GCF) MD-55-2, a radiochromic material, was examined for its optical properties through total reflectance and transmittance measurements in visible spectrum (400-700 nm). By using a multilayer model of the film and Kubelka-Munk's (KM) theory, absorption and scattering coefficients of the film sensitive layer (K and S, respectively) were obtained from measurements of irradiated and nonirradiated slides. This has allowed calculation of the absorbance A(KM) of the sensitive layer of the GCF. The model easily splits scattering from absorption. Unlike absorption, scattering is essentially insensitive to irradiation dose and decreases slowly as the wavelength increases. The scattering effect is predominant over absorption in the 400-500 nm range, while beyond 600 nm absorption prevails. The A(KM) absorbance of the sensitive layer was calculated using the K coefficient and compared with the optical densities (OD) measured considering only ballistic photons (as in a standard spectrophotometer) as well as the optical densities measured collecting all the transmitted photons (as in many densitometers). The values of A(KM) found were always lower than OD measured by the other methods and they had the best linearity on the whole visible range. These data support the hypothesis that the sensitive layer reacts to irradiation more linearly than that shown by measurements using standard commercial devices. However, in the 600-680 nm range, correction is not very important because absorption is predominant over scattering. When GCF is used for imaging, scattering produces a loss of spatial information. Consequently, it is necessary to collect only ballistic photons and to correct absorbance by K and S coefficients.     

Diffuse angiomatosis of the extremities presenting as a sarcoma

Diffuse angiomatosis of the extremities is a rare condition characterized by extensive or multifocal benign mass-forming vascular lesions involving multiple tissue planes; histologically, adipose and fibrous tissue are admixed with vascular elements that vary from capillary proliferations to large vessels that are often structurally abnormal. Although diffuse angiomatosis most likely represents a hamartomatous form of arteriovenous malformation, it behaves clinically as a benign but slowly progressive and unresectable neoplasm. We describe a case of diffuse angiomatosis of the lower extremity that presented clinically as a soft-tissue sarcoma.     

The total serum homocysteine as an indicator of vitamin B12 and folate status

Presented is a modification of an assay for total serum homocysteine (Hcy) in which the Hcy plus radioactive adenosine is converted enzymatically to labeled S-adenosylhomocysteine (AdoHcy). The modifications included a commerical source for the AdoHcy hydrolase, adenosine labeled with either 14C or 3H, and separation of the AdoHcy by thin layer chromatography. The assay was sensitive to 25 pmol. Hcy levels in sera from 18 controls ranged from 6.9 to 12.1 mumol/L with a mean of 9.1 and a SD of 1.5 mumol/L. The total serum Hcy was increased in vitamin B12 and folate deficiency. The level was high in congenital defects of vitamin B12 metabolism, blocking the methylation of Hcy regardless of the serum vitamin B12 levels, but was normal in the absence of tissue deficiency even if the serum vitamin B12 levels were low. The procedure has been found practical in two years of use and requires only 0.1 mL of serum.     

Microcirculation of a bulbar conjunctiva in calves with the total artificial heart: results of a long-term study

The aim of this work was to study the microcirculation in calves with total artificial heart (TAH) using the observation of the microcirculatory bed of bulbar conjunctiva. The examination were performed on 21 calves with total artificial heart and 8 controls. The microcirculatory bed was registered using the flash photographic method. In experimental calves after artificial heart implantation the microvasculature was studied two times in a week, to follow the pathophysiological changes in longitudinal periods until 100 days, between 100 and 200 days and after 200 days. For the evaluation we used the correlation analysis between arteriovenous diameter ratio (A/V ratio) and the values of central venous pressure (CVP). On the basis of our results we can conclude that during the survival period with TAH the morphological changes of conjunctival microvasculature occur. Our results showed the varicosities, serpentine-like forms and typical congestive changes of small vessels in bulbar conjunctiva. The correlation analysis showed the positive correlation between arteriovenous diameter ratio (A/V ratio) and central venous pressure (CVP) in the late phase of survival period between 100 to 200 days and over 200 days. The number of pathological changes of calves with artificial heart implantation.     

Diffuse reflectance spectrophotometry with visible light: comparison of four different methods in a tissue phantom

The purpose of the study was to compare algorithms of four methods (plus two modifications) for spectrophotometric haemoglobin saturation measurements. Comparison was made in tissue phantoms basically consisting of a phosphate buffer, Intralipid and blood, allowing samples to be taken for reference measurements. Three experimental series were made. In experiment A (eight phantoms) we used the Knoefel method and measured specific extinction coefficients with a reflection spectrophotometer. In experiment B (six phantoms) the fully oxygenated phantoms were gradually deoxygenated with baker's yeast, and simultaneous measurements were made with our spectrophotometer and with a reference oxymeter (ABL-605) in 3 min intervals. For each spectrophotometric measurement haemoglobin saturation was calculated with all algorithms and modifications, and compared with reference. In experiment C (11 phantoms) we evaluated the ability of a modification of the Knoefel method to measure haemoglobin concentration in absolute quantities using extinction coefficients from experiment A.Results. Experiment A: with the Knoefel method extinction coefficients (+/-SD) for oxyhaemoglobin at 553.04 and 573.75 nm were 1.117 (+/-0.0396) ODmM(-1) and 1.680 (+/- 0.0815) ODmM(-1), respectively, and for deoxyhaemoglobin 1.205 (+/- 0.0514) ODmM(-1) and 0.953 (+/-0.0487) ODmM(-1), respectively. Experiment B: high correlation with the reference was found in all methods (r = 0.94-0.97). However, agreement varied from evidently wrong in method 3 and the original method 4 (e.g. saturation above 160%) to high agreement in method 2 as well as the modifications of methods 1 and 4, where oxygen dissociation curves were close to the reference method. Experiment C: with the modified Knoefel method the mean haemoglobin concentration difference from reference was 8.3% and the correlation was high (r = 0.91).We conclude that method 2 and the modifications of 1 and 4 were superior to the others, but depended on known values in the same or similar phantoms. The original method 1 was independent of results from the tissue phantoms, but agreement was slightly poorer. Method 3 and the original method 4 could not be recommended. The ability of the modified method 1 to measure haemoglobin concentration is promising, but needs further development.     

Determination of the complex refractive index of highly concentrated hemoglobin solutions using transmittance and reflectance measurements

The complex refractive index of highly concentrated hemoglobin solutions as they appear in red blood cells are determined in the wavelength range of 250 to 1100 nm using transmittance and Fresnel reflectance measurements. The determined real parts of the refractive indices are on average 0.02 units higher than the values found in the literature. The wavelength dependence of the measured data in the UV region differs from the calculated data using the Kramers-Kronig relation.     

Ploidy as a prognostic indicator in oesophageal squamous carcinoma and its relationship to various histological criteria

One hundred surgically resected squamous cell carcinomas of the oesophagus were studied retrospectively to assess the significance of DNA aneuploidy as determined by flow cytometry using paraffin-embedded tissue. DNA aneuploidy was not an independent prognostic indicator but was found to be associated with tumour necrosis and host/tumour interface fibrosis. When tumours confined to submucosa or muscle wall were assessed, diploid tumours had a poorer survival rate than DNA aneuploid tumours. Other histological variables studied were tumour differentiation, depth of infiltration, glandular and small cell differentiation, lymphatic and vascular spread and host inflammatory response. These failed to show any significant association with ploidy.     

Peculiar character of dialyzate ultraviolet extinction spectra as an indicator of nucleic acid metabolism in humans

UV extinction spectra of effluent dialyzate in the process of hemodialysis and peritoneal dialysis are studied. It is found that they have distinctive individual features unique to each patient. The shape of the spectrum remains virtually invariable for a given subject during the whole period of the study (more than a year) and does not depend on the treatment modality or other factors like sex, age, the stage of chronic renal failure, and concomitant illnesses. Absorption bands in two spectral regions--255 to 265 nm and 288 to 298 nm--account for the unique character of dialyzate spectra. Spectra classification based on the ratio of integral extinctions in these regions are introduced. It is assumed that the shape of the spectrum depends on the relative contents of plasma components. Statistical analysis of individual features of spectra for the groups of patients undergoing hemodialysis (78 people) and peritoneal dialysis (42 people) is carried out. It is revealed that the distribution of patients by the ratio of integral extinctions in the specified spectral regions adheres to the normal law. Presumably individual features of spectra could be related to uric acid and a biochemically unidentified substance with an absorption peak near 260 nm.     

Microcirculation of the bulbar conjunctiva in the goat implanted with a total artificial heart: effects of pulsatile and nonpulsatile flow

A new system to observe the microcirculation on the bulbar conjunctiva was developed using a digital high definition microscope to investigate the influence of the flow patterns on the microcirculation in a goat with a total artificial heart (TAH). The undulation pump TAH was implanted into the goat. When the whole body condition became stable, the flow pattern was modulated between the pulsatile and the nonpulsatile mode, and the changes in the microcirculation were observed. When the flow pattern was changed from pulsatile to nonpulsatile mode, the erythrocyte velocity in capillaries dropped from 526+/-83 to 132+/-41 microm/s and remained at a low level. The number of perfused capillaries decreased as well. Then the nonpulsatile flow mode was maintained for 20 minutes. After the flow pattern was returned to the pulsatile mode again, the erythrocyte velocity recovered to the initial level (433+/-71 microm/s). In many cases, the flow of the nonperfused capillaries in the nonpulsatile mode recovered to the initial level after the flow pattern was changed to the pulsatile mode again. The perfused capillary density in the nonpulsatile mode (19.7+/-4.1 number of capillaries/mm) was significantly lower than that in the pulsatile mode (34.7+/-6.3 number of capillaries/mm). It is thought that the basal and flow stimulated endothelium derived nitric oxide release in the microvessels decreased because of the disappearance of pulsatility and that the nitric oxide induced the constriction of arterioles after the flow pattern was changed to the nonpulsatile mode. At the same time, the baroceptors might sense the decrease in the arterial peak pressure or dp/dt, and the sympathetic nerve increases activities and induce the constriction of arterioles. Then, the erythrocyte velocity in capillaries would decrease. Because of the flow pattern further in the chronic phase, it is important to follow the change in the microcirculation.