Gallstones: A Worldwide Multifaceted Disease and Its Correlations with Gallbladder Carcinoma

RESEARCH ARTICLE Gallstones: A Worldwide Multifaceted Disease and Its Correlations with Gallbladder Carcinoma Raj Kumar Sharma1,2, Kanchan Sonkar1,3, Neeraj Sinha1*, Pradeep Rebala4, Ahmad Ebrah Albani5, Anu Behari6, Duvvuri Nageshwar Reddy4, Alvina Farooqui2, Vinay Kumar Kapoor6 1 Centre of Biomedical Research, Sanjay Gandhi Post Graduate Institute of Medical Sciences (SGPGIMS) Campus, Lucknow, India, 2 Department of Biosciences, Integral University, Kursi road, Lucknow, India, 3 Department of Biochemistry & BiophysicsUniversity of Pennsylvania Perelman School of Medicine, Philadelphia, United States of America, 4 Department of Surgical Gastroenterology, Asian Institute of Gastroenterology, Somajiguda, Hyderabad, Andhra Pradesh, India, 5 Department of Surgery, Zayad Military Hospital, Abu Dhabi, UAE, 6 Department of Surgical Gastroenterology, SGPGIMS, Raibarelly Road Lucknow, Uttar Pradesh, India a11111 * , Abstract OPEN ACCESS Citation: Sharma RK, Sonkar K, Sinha N, Rebala P, Albani AE, Behari A, et al. (2016) Gallstones: A Worldwide Multifaceted Disease and Its Correlations with Gallbladder Carcinoma. PLoS ONE 11(11): e0166351. doi:10.1371/journal. pone.0166351 Editor: Tzen-Yuh Chiang, National Cheng Kung University, TAIWAN Received: June 27, 2016 Background Gallstones (GS) associated diseases are among the most recurrent and frequent diseases delineated in India and United Arab Emirates. Several reports suggest that the association of GS with gallbladder cancer (GBC) is very high in Northern part of India; however, its occurrence in UAE and Southern part of India is notably low. Therefore, in the present study, we aimed to perform compositional analysis of GS in three different geographical areas by Solid State Nuclear Magnetic Resonance and Fourier Transformed Infrared spectroscopy. Accepted: October 27, 2016 Published: November 10, 2016 Copyright: © 2016 Sharma et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Funding: The authors received no specific funding for this work. Competing Interests: The authors have declared that no competing interests exists. Methods Natural abundance 13C cross polarization magic angle spinning Nuclear Magnetic Resonance and Fourier Transform Infrared spectroscopy is employed for the analysis of human gallstone. Results Cholesterol, bilirubin and calcium carbonate were present in variant concentrations in GS obtained from three different geographical regions. Cholesterol was present predominantly in gallstones from North India. Bilirubin was found to be a main constituent in gallstones pertaining to South India. Whereas GS from UAE showed both cholesterol and bilirubin as their major constituents. Calcium carbonate was found in varying concentrations in gallstones acquired from different regions. PLOS ONE | DOI:10.1371/journal.pone.0166351 November 10, 2016 1 / 14 Analysis of Gallstone Using NMR and FTIR Spectroscopy Conclusion Variation in environmental condition and dietary habits may contribute and affect the GS formation. Alterations in bile composition influence the GB and augment the crystallization of cholesterol. Analysis of different geographical regions GS could be an important stride to understand the etiology of GS diseases. Introduction Gallstone (GS) disease is a very common health problem that affects millions throughout the world [1, 2]. GS are generally found in gallbladder (GB) which occupies the central part of human biliary system. GB acts as a reservoir of the bile fluid produced by the liver. Bile is concentrated in GB and drained out for lipid absorption. GB epithelium is most absorptive epithelium surface which facilitates bile concentration in GB by trans-mucosal absorption. Bile constitutes cholesterol, bilirubin, bile salts and phospholipids which are the key components to keep up the homeostasis of GB. There are various pathological conditions that can disturb the regulation of these secretions. Altered secretion of biliary cholesterol, phospholipids and bile acids disturbs the solubility of bile [3]. GS formation commences with cholesterol super saturation [4]. This corresponds to excess amount of cholesterol secretion or owing to less concentration of bile salts and phospholipids which are necessary for solubility of cholesterol [5, 6]. Secretion of cholesterol increases with age and correlated with dietary factors and fasting state. Cholesterol forms crystals that remains in the gall bladder and combines with other constituents like calcium salt, magnesium salts and bilirubin to form GS. GS occurs in various shapes (round, angular, oval and so on), sizes (from few millimeters to 6 cm), color (creamy white, yellow, black and brown) and can be amorphous or crystalline [7]. Worldwide distribution of GS has been described in several reports. There is notorious relationship with GS and GBC. Patients having history of GS are more susceptible to GBC. The most important risk factor for developing GBC is attributed to long standing period and GS size measuring greater than 3.0 cm [8]. Nevertheless, there is no proven evidence of association between GS and GBC. In accordance with the above mentioned context the composition of GS obtained from different environmental conditions and geographical regions may offer clues to the pathogenesis of GBC, therefore determining the composition of GS becomes very important. There are various techniques that can be employed for this purpose. These techniques include X-ray diffraction, fluorescent spectroscopy, Fourier transform infrared spectroscopy (FTIR) and Nuclear Magnetic Resonance (NMR) Spectroscopy. These techniques have contributed meaningful insight and information. NMR spectroscopy has applicability to analyze biological samples in disease conditions with minimal sample preparation and reproducibility. Both solid and solution form of samples can be analyzed by NMR spectroscopy. For using NMR in solution state, sample is dissolved in a suitable solvent and for solid state; sample can be used as powder, solid or crystalline form. 13 C CP-MAS provides complete structure analysis of organic molecule as it takes into consideration of all organic constituents in GS both soluble and non-soluble. FTIR spectroscopy has been used as an important technique for characterizing biomolecules. Quick results make it an excellent technique. FTIR is important to obtain information about organic and inorganic constituents of GS. It gives clear absorption peaks for organic and inorganic constituents. PLOS ONE | DOI:10.1371/journal.pone.0166351 November 10, 2016 2 / 14 Analysis of Gallstone Using NMR and FTIR Spectroscopy In the present article we report analysis of GS from (...truncated)