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1.
Molecules ; 25(14)2020 Jul 21.
Artículo en Inglés | MEDLINE | ID: mdl-32708082

RESUMEN

Lymph nodes (LNs) play a very important role in the spread of cancer cells. Moreover, it was noticed that the morphology and chemical composition of the LNs change in the course of cancer development. Therefore, finding and monitoring similarities between these characteristics of the LNs and tumor tissues are essential to improve diagnostics and therapy of this dreadful disease. In the present study, we used Raman and Fourier transform infrared (FTIR) spectroscopies to compare the chemical composition of the breast cancer tissues and LNs collected from women without (I group-4 patients) and with (II group-4 patients) recurrence. It was shown that the similarity of the chemical composition of the breast tissues and LNs is typical for the II group of the patients. The average Raman spectrum of the breast cancer tissues from the I group was not characterized by vibrations in the 800-1000 cm-1 region originating from collagen and carbohydrates, which are typical for tumor-affected breast tissues. At the same time, this spectrum contains peaks at 1029 cm-1, corresponding to PO2- from DNA, RNA and phospholipids, and 1520 cm-1, which have been observed in normal breast tissues before. It was shown that Raman bands of the average LN spectrum of the II group associated with proteins and carbohydrates are more intensive than those of the breast tissues spectrum. The intensity of the Raman spectra collected from the samples of the II group is almost three times higher compared to the I group. The vibrations of carbohydrates and amide III are much more intensive in the II group's case. The Raman spectra of the breast cancer tissues and LNs of the II group's samples do not contain bands (e.g., 1520 cm-1) found in the Raman spectra of the normal breast tissues elsewhere. FTIR spectra of the LNs of the I group's women showed a lower level of vibrations corresponding to functional group building nucleic acid, collagen, carbohydrates, and proteins in comparison with the breast cancer tissues. Pearson's correlation test showed positive and more significant interplay between the nature of the breast tissues and LN spectra obtained for the II group of patients than that in the I group's spectra. Moreover, principal component analysis (PCA) showed that it is possible to distinguish Raman and FTIR spectra of the breast cancer tissues and LNs collected from women without recurrence of the disease.


Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Mama/química , Mama/química , Ganglios Linfáticos/química , Anciano , Anciano de 80 o más Años , Mama/citología , Carbohidratos/análisis , ADN/análisis , Femenino , Humanos , Ganglios Linfáticos/citología , Persona de Mediana Edad , Recurrencia Local de Neoplasia/química , Fosfolípidos/análisis , Análisis de Componente Principal , Proteínas/análisis , ARN/análisis , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Espectrometría Raman/métodos
2.
Biosensors (Basel) ; 9(1)2019 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-30823455

RESUMEN

We registered surface enhanced Raman scattering (SERS) spectra of the human lactoferrin molecules adsorbed on a silvered porous silicon (por-Si) from 10-6⁻10-18 M solutions. It was found that the por-Si template causes a negative surface potential of silver particles and their chemical resistivity to oxidation. These properties provided to attract positively charged lactoferrin molecules and prevent their interaction with metallic particles upon 473 nm laser excitation. The SERS spectra of lactoferrin adsorbed from 10-6 M solution were rather weak but a decrease of the concentration to 10-10 M led to an enormous growth of the SERS signal. This effect took place as oligomers of lactoferrin were broken down to monomeric units while its concentration was reduced. Oligomers are too large for a uniform overlap with electromagnetic field from silver particles. They cannot provide an intensive SERS signal from the top part of the molecules in contrast to monomers that can be completely covered by the electromagnetic field. The SERS spectra of lactoferrin at the 10-14 and 10-16 M concentrations were less intensive and started to change due to increasing contribution from the laser burned molecules. To prevent overheating the analyte molecules on the silvered por-Si were protected with graphene, which allowed the detection of lactoferrin adsorbed from the 10-18 M solution.


Asunto(s)
Técnicas Biosensibles , Grafito/química , Lactoferrina/aislamiento & purificación , Espectrometría Raman/métodos , Humanos , Lactoferrina/química , Porosidad , Silicio/química , Plata/química , Propiedades de Superficie
3.
Arch Biochem Biophys ; 638: 66-75, 2018 01 15.
Artículo en Inglés | MEDLINE | ID: mdl-29273432

RESUMEN

Investigations of short peptides that can be used in the next phase of synthetic HIV1 vaccine development are an urgent goal, as well as investigations of peptides that can be used in immunological tests with the aim to check the titer of antibodies against the alpha helix 1 from the first conserved region of HIV1 gp120 that are known to cause antibody-dependent cellular cytotoxicity (ADCC). The aim of this work was to study the structure of the NQ21 peptide corresponding to the less mutable part of the first conserved region of HIV1 gp120 (residues 94-114). The NQ21 peptide and its conjugate with biotin (biotin-NQ21) are absolutely alpha-helical in phosphate buffer solutions at pH = 6.8, 7.4 and 8.0, as well as in the dried form, according to the results of surface-enhanced Raman scattering (SERS) spectroscopy. Results of the native gel electrophoresis and thermal analysis under the control of spectrofluorometer and near UV circular dichroism (CD) showed that the peptide exists in form of octamers and tetramers at pH = 7.4, that is important information for further vaccine development. Strong signal of interacting Trp residues in oligomers in the far UV CD obscures the signal from secondary structure, but becomes less intensive during the heating.


Asunto(s)
Proteína gp120 de Envoltorio del VIH/química , VIH-1/química , Péptidos/química , Vacunas contra el SIDA/química , Dicroismo Circular , Concentración de Iones de Hidrógeno , Estructura Secundaria de Proteína , Espectrometría Raman
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