Comparison of surface enhanced Raman spectroscopy and Raman spectroscopy for the detection of breast cancer based on serum samples.

In this study, surface enhanced Raman spectroscopy (SERS) and Raman spectroscopy (RS), are employed for the classification of different stages of breast cancer using clinically diagnosed serum samples from breast cancer patients and healthy individuals. These serum samples are compared for their spectral features acquired by SERS and RS to establish spectral features that can be considered as spectral markers of breast cancer diagnosis and classification. SERS features related to DNA, proteins and lipids were observed which are solely observed in the serum samples of patients at different stages of breast cancer as compared to healthy samples. In order to explore the capability of SERS and RS and their comparison as an analytical tool for the efficient understanding of the progression of breast cancer, Principal Component Analysis (PCA) is done for the SERS and RS spectra of control, stage 2, stage 3 and stage 4. Furthermore, the Partial Least Squares-Discriminant Analysis (PLS-DA) was performed to compare the diagnostic performance of SERS and Raman spectroscopy for the classification of disease positive samples and healthy ones. The sensitivity and specificity and area under receiver operating characteristic (AUROC) curve values for SERS data were 90%, 98.4%, and 94% respectively which were higher as compared to Raman spectral data for which these values were found to be 88.2%, 97.7%, and 83.4% respectively.

Raman spectroscopy of blood plasma samples from breast cancer patients at different stages.

Raman spectroscopy was employed for the characterization of blood plasma samples from patients at different stages of breast cancer. Blood plasma samples taken from clinically diagnosed breast cancer patients were compared with healthy controls using multivariate data analysis techniques (principal components analysis - PCA) to establish Raman spectral features which can be considered spectral markers of breast cancer development. All the stages of the disease can be differentiated from normal samples. It is also found that stage 2 and 3 are biochemically similar, but can be differentiated from each other by PCA. The Raman spectral data of the stage 4 is found to be biochemically distinct, but very variable between patients. Raman spectral features associated with DNA and proteins were identified, which are exclusive to patient plasma samples. Moreover, there are several other spectral features which are strikingly different in the blood plasma samples of different stages of breast cancer. In order to further explore the potential of Raman spectroscopy as the basis of a minimally invasive screening technique for breast cancer diagnosis and staging, PCA-Factorial Discriminant Analysis (FDA) was employed to classify the Raman spectral datasets of the blood plasma samples of the breast cancer patients, according to different stages of the disease, yielding promisingly high values of sensitivity and specificity for all stages.

Gamma-interferon induces apoptosis of the B lymphoma WEHI-279 cell line through a CD95/CD95L-independent mechanism.

Gamma-interferon (IFN-gamma) a cytokine produced by CD4+ T helper type 1 cells, CD8+ T cells and natural killer (NK) cells, plays a central role in the development of humoral and cell-mediated immunity. IFN-gamma participates in the maturation and differentiation of B cells, but it has been previously reported that IFN-gamma may inhibit the early stages of B cell activation. We report that the inhibition of the B lymphoma cell WEHI-279-proliferation induced by IFN-gamma, involves the induction of typical features of apoptosis (nuclear chromatin condensation and fragmentation, cell shrinkage, phosphatidyl-serine (PS) exposure and mitochondrial membrane potential (delta psim) loss). IFN-gamma-mediated B cell apoptosis was decreased by the addition of the T helper type 2 cytokine, IL-4. WEHI-279 cells express CD95 and undergo apoptosis after treatment with either an agonistic anti-CD95 Ab or with a soluble recombinant CD95L. However, incubation with CD95-Fc or TRAIL-R1-Fc fusion proteins, did not prevent IFN-gamma-mediated apoptosis, suggesting that IFN-gamma-mediated apoptosis occurs independently of CD95/CD95L and TRAIL-R/TRAIL interactions. IFN-gamma-mediated apoptosis is associated with caspase-3 activation that can be prevented by the addition of the broad caspase inhibitor zVAD-fmk. These data indicate that IFN-gamma may play a major role in the regulation of B cell apoptosis, and suggest the involvement of an alternative pathway which is independent of the death receptors.

Phosphorylation of Bcl-2 and mitochondrial changes are associated with apoptosis of lymphoblastoid cells induced by normal immunoglobulin G.

Normal human immunoglobulin G induces apoptosis in human lymphoblastoid cells which involves antibody-mediated Fas ligation and the activation of caspases. Here, we show that Bcl-2 is phosphorylated on serine upon treatment of CEM T cells with normal IgG and that the overexpression of Bcl-2 in stable transfectants of CEM T cells prevents IgG-induced cell death. Treatment of CEM cells with normal IgG also results in a reduction in mitochondrial transmembrane potential and in the release of cytochrome c (Cyt c) into cytosol. The findings are concordant with earlier observations that apoptosis induced by IgG is associated with the activation of caspases. Our results demonstrate that Bcl-2 controls apoptosis induced by normal IgG and support a central role for Bcl-2 and mitochondria in antibody-mediated selection of lymphocyte repertoires.