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Universal health care is attracting increased attention nowadays, because of the large increase in population all over the world, and a similar increase in life expectancy, leading to an increase in the incidence of non-communicable (various cancers, coronary diseases, neurological and old-age-related diseases) and communicable diseases/pandemics like SARS-COVID 19. This has led to an immediate need for a healthcare technology that should be cost-effective and accessible to all. A technology being considered as a possible one at present is liquid biopsy, which looks for markers in readily available samples like body fluids which can be accessed non- or minimally- invasive manner. Two approaches are being tried now towards this objective. The first involves the identification of suitable, specific markers for each condition, using established methods like various Mass Spectroscopy techniques (Surface-Enhanced Laser Desorption/Ionization Mass Spectroscopy (SELDI-MS), Matrix-Assisted Laser Desorption/Ionization (MALDI-MS), etc., immunoassays (Enzyme-Linked Immunoassay (ELISA), Proximity Extension Assays, etc.) and separation methods like 2-Dimensional Polyacrylamide Gel Electrophoresis (2-D PAGE), Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), Capillary Electrophoresis (CE), etc. In the second approach, no attempt is made the identification of specific markers; rather an efficient separation method like High-Performance Liquid Chromatography/ Ultra-High-Performance Liquid Chromatography (HPLC/UPLC) is used to separate the protein markers, and a profile of the protein pattern is recorded, which is analysed by Artificial Intelligence (AI)/Machine Learning (MI) methods to derive characteristic patterns and use them for identifying the disease condition. The present report gives a summary of the current status of these two approaches and compares the two in the use of their suitability for universal healthcare.

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Introduction: The worldwide increase in the prevalence of obesity over the years has emerged as a global health concern. The growing rate of obesity in women of child bearing age is particularly a matter of concern. Obesity is considered a risk factor that predisposes an individual to a proinflammatory state through the release of the inflammatory mediators. Recent studies have shown a positive correlation between the severity of inflammation and an increase in adenosine deaminase (ADA) and high sensitivity C- reactive protein (hs-CRP). Obese pregnancy women are at a higher risk for developing inflammation-mediated pregnancy complications like gestational diabetes, preeclampsia, and preterm delivery. Considering the fact that pregnancy, obesity and inflammation are closely linked, this study evaluated the inflammation associated with obesity during pregnancy by estimating changes in ADA and hs-CRP. Materials and methods: The current study aimed to evaluate the levels of inflammation in obese pregnant women compared to non-obese pregnant women by correlating BMI with levels of ADA / hs-CRP. The study also aimed to examine the change in ADA and hs-CRP levels with gestational age (between the 1st and the 3rd trimester) in obese pregnant women as compared to non-obese pregnant women. We also examined whether changes in the levels of ADA correlate with changes in the levels of hs-CRP particularly in obese pregnant women.Blood samples were collected from obese and non-obese pregnant women. ADA activity and hs-CRP levels were estimated by biochemical assays. BMI was evaluated in the 1st trimester and those women with BMI > 30 kg/m2were considered as obese. Thirty subjects were included in each of the two groups. Results: ADA and hs-CRP levels were significantly higher in obese pregnant women in both the 1st and 3rd trimesters compared to non-obese participants (P value<0.05). Statistically significant higher values of ADA and hs-CRP were seen in obese participants in the 3rd trimester compared to the 1st trimester.A significant linear positive correlation was found between BMI and 3rd trimester ADA, and a linear positive correlation between BMI and hs-CRP both in the 1st and 3rd trimester. The relationship between ∆ ADA and ∆ hs-CRP was non- significant. Conclusions: The observations of this study reveal increased inflammatory responses in obese pregnant women and suggests the importance of ADA and hs-CRP as early indicators of obesity-related complications prevailing thereafter, these markers can be useful for clinical diagnosis of impending maternal and neonatal complications.

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BACKGROUND: Breast cancer is one of the most common cancers in India. Most of the patients with breast cancer are treated with chemotherapy which has multiple oral complications. AIMS: The objectives of this study were to describe the occurrence of taste disturbances, xerostomia, oral mucositis, oral pigmentation, and candidal and salivary changes among patients receiving chemotherapy for breast cancer. METHODS: Fifty-two women with newly diagnosed breast cancer (without distant metastasis), eligible for adjuvant/neoadjuvant chemotherapy (cyclophosphamide and adriamycin, 4 cycles × 3 weeks), were included in this study. All the observations were noted before, during (after 6 weeks of starting chemotherapy), and after the completion of chemotherapy (after 12 weeks of starting chemotherapy). STATISTICAL ANALYSIS USED: Variables such as mucositis, salivary flow rate, salivary pH, and candidal carriage rate were compared at baseline, and at 1st and 2nd follow-ups using Wilcoxon signed-rank test (P value corrected for α for pair-wise comparisons). RESULTS: Mean unstimulated whole salivary flow rate reduced from 0.5 ml/min to 0.3 ml/min, and the mean colony-forming units of Candida reduced from 32.3 × 103 cells/ml to 13.1 × 103 cells/ml at the end of the study period. Xerostomia, taste disturbances, and oral mucosal pigmentation increased from 28.8% to 50%. CONCLUSIONS: There was a discernible change in oral mucosal, salivary, and candidal status during the course of the study.

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AIMS AND OBJECTIVE: The purpose of the present study was to evaluate and compare flexural strength and Staphylococcus aureus adhesion of heat-activated poly (methyl methacrylate [MMA]) resin modified with a comonomer of methacrylic acid (MAA) and MMA monomer. MATERIALS AND METHODS: Comonomer preparation was done with the addition of varying concentration of MAA (0, 15, 20, and 25 wt %) to the MMA of conventional heat-activated denture base resin to prepare the specimens. Prepared specimens were stored in distilled water at 37°C for 1 day and 1 week before the evaluation of flexural strength and microbial adhesion. Flexural strength was measured using a universal testing machine at a crosshead speed for 2 mm/min (n = 10). Microbial adhesion (colony-forming unit [CFU]) was evaluated against S. aureus using a quadrant streaking method (n = 5). Data were subjected to one-way ANOVA, and the significant differences among the results were subjected to Tukey's HSD test. P < 0.05 was considered statistically significant. RESULTS: Addition of MAA to the MMA monomer was found to significantly reduce the adhesion of S. aureus for all the groups. Reduction of CFU of S. aureus was found be more significant for Group 3 as compared to control, both at 1-day (P < 0.001) and 1-week (P < 0.002) storage in distilled water. However, no statistically significant changes in the flexural strength were observed with the addition of MAA at 1-day (P = 0.52) and 1-week (P = 0.88) time interval. CONCLUSION: Addition of MAA to conventional denture base resin reduced the microbial adhesion without significantly affecting the flexural strength.

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An ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique has been developed by our group at Manipal, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from volunteers (normal, and different pre-malignant/malignant conditions) were recorded using this set-up. The protein profiles were analyzed using principal component analysis (PCA) to achieve objective detection and classification of malignant, premalignant and healthy conditions with high sensitivity and specificity. The HPLC-LIF protein profiling combined with PCA, as a routine method for screening, diagnosis, and staging of cervical cancer and oral cancer, is discussed in this paper. BIOLOGICAL SIGNIFICANCE: In recent years, proteomics techniques have advanced tremendously in life sciences and medical sciences for the detection and identification of proteins in body fluids, tissue homogenates and cellular samples to understand biochemical mechanisms leading to different diseases. Some of the methods include techniques like high performance liquid chromatography, 2D-gel electrophoresis, MALDI-TOF-MS, SELDI-TOF-MS, CE-MS and LC-MS techniques. We have developed an ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from healthy and volunteers with different malignant conditions were recorded by using this set-up. The protein profile data were analyzed using principal component analysis (PCA) for objective classification and detection of malignant, premalignant and healthy conditions. The method is extremely sensitive to detect proteins with limit of detection of the order of femto-moles. The HPLC-LIF combined with PCA as a potential proteomic method for the diagnosis of oral cancer and cervical cancer has been discussed in this paper. This article is part of a Special Issue entitled: Proteomics in India.

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Cervical cancer, the second most common cancer in women, progresses silently over long periods before producing any clinical manifestation. Research in early detection of this condition using proteomic techniques is of very recent origin. We used high-performance liquid chromatography combined with laser-induced fluorescence method in our lab to record the protein profiles of tissue homogenate, cell lysate and serum samples of normal and different stages of malignant conditions of the cervix. Information on protein markers in the protein profiles was derived using various data processing methods including curve resolution. The variations in relative intensities of different peaks with respect to peak height, width and area under the curve from different sample types were compared to get information regarding the concentration of the various proteins and their significance in the induction and metastasis of cervical cancer. The method can be used in diagnosis, follow-up with respect to the progression, remission and effective therapy, in cervical malignancy.

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A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF) technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA) to get statistical information on different tissue components. Discrimination of different stages of the samples was carried out by considering three parameters--scores of factor, spectral residual, and Mahalanobis Distance. Diagnostic accuracy of the method was evaluated using Receiver Operating Characteristic (ROC) analysis, and Youden's index (J) plots. The PCA results showed high sensitivity and specificity (~100) for cervical cancer diagnosis. ROC and Youden's index curves for both normal and malignant standard sets show good diagnostic accuracy with high AUC values. The statistical analysis has shown that the differences in protein profiles can be used to diagnose biochemical changes in the tissue, and thus can be readily applied for the detection of cervical cancer, even in situations where a histopathology examination is not easy because of nonavailability of experienced pathologists.

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Protein profiles of cytologic samples from the cervix were studied using High Performance Liquid Chromatographic (HPLC) separation combined with ultra-sensitive laser induced fluorescence (LIF) detection. HPLC-LIF protein profiles of samples from clinically normal subjects, individuals suffering from cervical cancer (different stages), and subjects who had other gynecological problems related to cervix, like erosion of cervix and Nabothian cyst, but no malignancy, were subjected to Principal Component Analysis (PCA). The application of HPLC-LIF protein profiling combined with PCA was found to be a highly efficient method for discrimination of different classes of samples with high sensitivity and specificity. Diagnostic accuracy and optimal threshold - decision criterion - for objective discrimination were estimated using sensitivity-specificity pairs and Youden's index (J) plots.

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The present work deals with the evaluation of a high-performance liquid chromatography laser-induced fluorescence (HPLC-LIF) technique developed in our laboratory for early detection of oral cancer from protein profiles of body fluids. The results show that protein profiles of serum samples from a given class of samples, say, normal, premalignant, or malignant, are statistically very close to each other, while profiles of members of any class are significantly different from other classes. The performance of the technique is evaluated by the use of sensitivity and specificity pairs, receiver operating characteristic (ROC) analysis, and Youden's Index. The technique uses protein profile differences in serum samples, registered by the HPLC-LIF technique. The study is carried out using serum samples from volunteers diagnosed as normal or premalignant clinically, and as malignant by histopathology. The specificities and sensitivities of the HPLC-LIF method at an ideal threshold (M-distance = 2) for normal, malignant, and premalignant classes are 100, 69.5, and 61.5%, and 86.5, 87.5, and 87.5% respectively.

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