Background: Hypoxia-induced CA IX contributes to pH control in tumor cells, and control of pH is important for cell migration. Results: CA IX increases migration through catalytic domain and interacts with bicarbonate transporters in lamellipodia. Conclusion: CA IX is an active component of the molecular machinery that facilitates migration of tumor cells through pH regulation at the leading edge membranes. Significance: This identifies CA IX as a target to suppress cell migration and reduce tumor aggressiveness.
In this contribution, we present a novel quadruple wavelength infrared sensor system for measuring the CH2-symmetric/CH2-antisymmetric stretch ratio of biological cells. This ratio can be used as a marker to distinguish between healthy and carcinoma cells. Compared to common techniques our system does not require specific labelling or staining, or the use of expensive liquid nitrogen cooled IR spectrometers. Two of the four wavelengths, 3.33 and 3.57 µm, are used as reference points and the other two represent CH2-symmetric and CH2-antisymmetric stretch absorption (respectively at 3.51 and 3.42 µm). IR absorbance spectra recordings of healthy (MDCK) and malignant (Caki-1) epithelial kidney cells with a conventional IR spectroscope showed significant differences in the absorbance ratio 3.51 µm/3.42 µm (CH2-symmetric/CH2-antisymmetric stretch). The sensor has been validated by measuring the CH2 stretch ratio of yeast samples both by IR spectroscopy and the sensor system. The methods yielded similar results. The application potential of our system is demonstrated by CH2 stretch measurements on healthy and carcinoma epithelial kidney cells. Measurements on both cell types yielded significant differences in CH2 stretch ratio. The sensor has the potential to be further developed into a fast, low-cost and label-free screening system for suspicious biopsy samples.
In this paper, we assess the potential of a label-free infrared absorbance based measurement method for determination of the CH(2)-symmetric to CH(2)-antisymmetric stretch ratio, to aid in the detection of the presence of cancer cells and to differentiate between various cancer cells. For this study a normal epithelial kidney cell line, two carcinoma epithelial kidney cell lines, an adult primary human melanocyte cell line, and three human melanoma cell lines were investigated. For the measurements we used a self-designed IR sensor which has the potential to be further developed in a point-of-care instrument. To investigate the mechanism influencing the CH(2)-stretch ratio of mammalian cell membranes, a normal epithelial kidney cell line was exposed to the plasma membrane bound cholesterol reducing agent methyl-β-cyclodextrin. This methodology yielded statistically significant CH(2)-stretch ratio differences between the individual cell lines, normal and tumorous, of both epithelial kidney and melanocyte origin. Measurement results of normal epithelial kidney cells exposed to methyl-β-cyclodextrin indicate that an increase in the CH(2)-stretch ratio arises when there is a decrease in, or redistribution of, the membrane stabilizing agent cholesterol. This study proves that the proposed cell type discrimination method, based on the CH(2)-symmetric to CH(2)-antisymmetric stretch ratio, allows the discrimination between normal and tumor cells. In addition, the method shows high potential for improvement of staging of suspicious tissues.
The application of chromosome banding and the new molecular cytogenetics techniques, in particular fluorescent in situ hybridization (FISH) using chromosome painting probes found early use for understanding the systematic and phylogenetic relationships of species, as well as for providing insights to the possible mechanisms underpinning speciation. Here, we showed an application of primed in situ DNA synthesis (PRINS) and fluorescent in situ hybridization for comparative detection of the domestic horse (Equus caballus) and the domestic donkey (Equus asinus) telomere, centromere and nucleolar-organizer region sequences. The number, distribution and kind of highly repeated DNA sequences in across species with regard to their advantage and limit usefulness in phylogenetic analyses are discussed.
Animal fats are considered to be unhealthy, in contrast to vegetable fats, which are rich in unsaturated fatty acids. However, the use of some fats, such as coconut oil, is still controversial. In our experiment, we divided experimental animals (domestic pigs) into three groups differing only in the type of fat used in the diet: group R: rapeseed oil (n = 5); group B: beef tallow (n = 5); group C: coconut oil (n = 6). After transcriptomic analysis of liver samples, we identified 188, 93, and 53 DEGs (differentially expressed genes) in R vs. B, R vs. C, and B vs. C comparisons, respectively. Next, we performed a functional analysis of identified DEGs with String and IPA software. We observed the enrichment of genes engaged in the unfolded protein response (UPR) and the acute phase response among genes upregulated in B compared to R. In contrast, cholesterol biosynthesis and cholesterol efflux enrichments were observed among genes downregulated in B when compared to R. Moreover, activation of the UPR and inhibition of the sirtuin signaling pathway were noted in C when compared to R. The most striking difference in liver transcriptomic response between C and B was the activation of the acute phase response and inhibition of bile acid synthesis in the latest group. Our results suggest that excessive consumption of animal fats leads to the activation of a cascade of mutually propelling processes harmful to the liver: inflammation, UPR, and imbalances in the biosynthesis of cholesterol and bile acids via altered organelle membrane composition. Nevertheless, these studies should be extended with analysis at the level of proteins and their function.
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