“…The intensity of this peak is high in S and low in P, and may be attributed to majority of salivary proteins present in supernatant phase (S) upon centrifugation; while the pellet (P) contains few proteins that settle down on the basis of weight. 34 This indicates that some molecules could be present in both phases, but at different concentrations. Even on cursory look one can observe that spectra of D and L forms are quite similar, and S and P present very different pattern suggesting that biochemical composition of D and L forms is less prone to sample processing unlike S and P. Figure 2.Mean Raman spectra of air dried, lyophilized, pellet, and supernatant saliva.…”
Minimally invasive cancer detection using bio-fluids is been actively pursued due to practical limitations of, though, better suited noninvasive and online in vivo methods. Saliva is one such clinically informative bio-fluid that offers advantage of easy and multiple sample collection. Despite its potential in cancer diagnostics, saliva analysis is challenging due to its heterogeneous composition. Recently, there has been an upsurge in saliva exploration by optical techniques. Although forms of saliva such as precipitate and supernatant have been monitored, but due to obvious loss of analytes in processing, sampling method needs to be standardized. In that context, present work details the comparison of four different saliva sampling methodologies i.e., air-dried (D), lyophilized (L), pellet (P) and supernatant (S) by Raman spectroscopy (RS) collected from 10 healthy samples. Composition driven spectral features of all forms were compared and classified using principal component analysis (PCA) and linear discriminant analysis (PCLDA). Analysis was carried out on all four groups in the first step. In the second step groups of P and S; and D and L were analyzed. Findings suggest that P and S exhibit discrete spectroscopic features and demonstrated high classification efficiency which is indicative of their distinctive biochemical composition. On the other hand, D and L forms showed overlapping spectral features and low classification suggesting these forms retain majority spectroscopic features of whole saliva and less prone to sampling losses. Thus, this study indicates D and L forms may be more appropriate for saliva sampling by RS providing the comprehensive information, required for cancer diagnosis. Further the method was also tested for classification of oral cancer and healthy subjects (n= 27) which yields 90 % stratification. Thus the findings of the study indicate utility of minimally invasive salivary Raman based diagnostic techniques in oral cancers.
“…The intensity of this peak is high in S and low in P, and may be attributed to majority of salivary proteins present in supernatant phase (S) upon centrifugation; while the pellet (P) contains few proteins that settle down on the basis of weight. 34 This indicates that some molecules could be present in both phases, but at different concentrations. Even on cursory look one can observe that spectra of D and L forms are quite similar, and S and P present very different pattern suggesting that biochemical composition of D and L forms is less prone to sample processing unlike S and P. Figure 2.Mean Raman spectra of air dried, lyophilized, pellet, and supernatant saliva.…”
Minimally invasive cancer detection using bio-fluids is been actively pursued due to practical limitations of, though, better suited noninvasive and online in vivo methods. Saliva is one such clinically informative bio-fluid that offers advantage of easy and multiple sample collection. Despite its potential in cancer diagnostics, saliva analysis is challenging due to its heterogeneous composition. Recently, there has been an upsurge in saliva exploration by optical techniques. Although forms of saliva such as precipitate and supernatant have been monitored, but due to obvious loss of analytes in processing, sampling method needs to be standardized. In that context, present work details the comparison of four different saliva sampling methodologies i.e., air-dried (D), lyophilized (L), pellet (P) and supernatant (S) by Raman spectroscopy (RS) collected from 10 healthy samples. Composition driven spectral features of all forms were compared and classified using principal component analysis (PCA) and linear discriminant analysis (PCLDA). Analysis was carried out on all four groups in the first step. In the second step groups of P and S; and D and L were analyzed. Findings suggest that P and S exhibit discrete spectroscopic features and demonstrated high classification efficiency which is indicative of their distinctive biochemical composition. On the other hand, D and L forms showed overlapping spectral features and low classification suggesting these forms retain majority spectroscopic features of whole saliva and less prone to sampling losses. Thus, this study indicates D and L forms may be more appropriate for saliva sampling by RS providing the comprehensive information, required for cancer diagnosis. Further the method was also tested for classification of oral cancer and healthy subjects (n= 27) which yields 90 % stratification. Thus the findings of the study indicate utility of minimally invasive salivary Raman based diagnostic techniques in oral cancers.
“…The sediment is an impurity in the form of residual reagents which unreacted in the formation of AgNPs so they are deposited due to the higher speed of centrifugation [22]. As a result of these collisions, the impurities are destabilized and form sediments.…”
Section: Figure 1 Color Of Agnps Solution Before and After The Centri...mentioning
The development of nanotechnology applications is rapidly growing in many sectors. One of them is silver nanoparticles (AgNPs) which are metal nanoparticles that play an important role, especially in nanomedicine. The most effective method of purifying to obtain stable AgNPs is very important to study. Experiments on the separation of AgNPs have been carried out using the size exclusion chromatography and centrifugation methods to see the effectiveness of refining the two methods. This experiment begins with the synthesis of AgNPs using the chemical reduction method. Then, the synthesized AgNPs were purified by Size Exclusion Chromatography (SEC) and centrifugation method then analyzed using UV-Vis spectrophotometer to determine the maximum peaks before and after purification. The experimental results were obtained that centrifugation methods and SEC having the same effectiveness in refining AgNPs. The centrifugation method at various speed (0, 3000, 6000, 9000, 12000 and 15000 rpm) gave wavelength results 403, 404, 404, 405, 404, and 404 nm. The SEC method using Sephadex-25 column showed the 4th to 8th fractions gave the maximum wavelength 404, 404, 404, 405, and 404 nm, respectively. The maximum wavelength of both methods showed the surface plasmon resonance characteristic of AgNPs. However, centrifugation at 3000 rpm has better homogeneity than SEC method.
“…Moreover, higher centrifugation speeds aid in the elimination of smaller proteins in the supernatant. Increasing the centrifugation speed from 8,000 g to 9,000 g results in the removal of larger proteins within the supernatant, particularly those with sizes of 25-150 kDa (Baskoro, et al, 2017). However, during the Pair T-Test analysis, it was statistically determined that the decrease in protein content following the centrifugation process did not achieve significant difference (p=0.246 or p>0.05).…”
An integral aspect of anticancer experimentation involves delineating the optimal dosage of the test compound to ascertain its efficacy in targeting malignant cells. Numerous variables may influence a compound's cytotoxicity, among which is the choice of cell culture medium. Within in vitro settings, supplementary mediums are employed to foster cellular proliferation. Platelet lysate (PL) serves as a growth supplement, presenting an alternative to fetal bovine serum (FBS), primarily due to its incorporation of growth factors such as platelet-derived growth factor (PDGF), a component absents in FBS. The integrity of PL may be subject to various factors, including the age of the donor. This study sought to evaluate the impact of donor age on PL quality. Furthermore, it aimed to discern whether PL derived from platelet concentrate (PC) blood components of different age cohorts influences the IC50 value in anticancer compound assessment. Expired PCs were utilized, subsequently classified into age categories: ≤30 years, >30 years, and a combination of ages. PL analysis encompassed parameters such as pH, blood profile, protein, glucose, and cholesterol levels. The investigation scrutinized the influence of PL quality, as a cellular growth supplement, on the anticancer compound cisplatin's activity against HeLa cells. Findings indicate that donor age influenced the IC50 value of cisplatin on HeLa cells. Notably, elevated cholesterol levels and decreased pH in PL from donor ages >30 years were associated with reduced cisplatin toxicity.Keywords: Cisplatin, Donor Age, HeLa, IC50, Platelet Lysate.
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