Nuclear Instruments and Methods in Physics Research B 267 (2009) 2878–2883

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Nuclear Instruments and Methods in Physics Research B

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Analysis of blood and tissue in gallbladder cancer

T.R. Rautray a,*, V. Vijayan b, M. Sudarshan c, S. Panigrahi d

a Department of Dental Biomaterials, School of Dentistry, Kyungpook National University, 2-188-1 Samduk-dong, Jung-gu, Daegu, Republic of Koreab Dept. of Physics, Valliammai Engineering College, SRM Nagar, Chennai, Indiac UGC-DAE Consortium for Scientific Research, Kolkata Centre, 3/LB-8 Bidhan Nagar, Kolkata 700 098, West Bengal, Indiad Dept. of Physics, National Institute of Technology, Rourkela 769 008, Orissa, India

a r t i c l e i n f o

Article history:Received 16 March 2009Received in revised form 30 May 2009Available online 18 June 2009

PACS:87.64.Ni87.68.+z87.64.Gb78.30.Er65.60.+a

Keywords:PIXEPIGETrace elementsGallbladder

0168-583X/$ - see front matter � 2009 Elsevier B.V.doi:10.1016/j.nimb.2009.06.084

* Corresponding author. Tel.: +82 536606897; fax:E-mail address: tapash77@hotmail.com (T.R. Rautr

a b s t r a c t

Particle induced X-ray emission, particle induced c-ray emission studies has been carried out to analysenormal and carcinoma tissues and blood samples of gallbladder of both sexes and seventeen trace ele-ments namely Na, Mg, Al, K, Ca, Ti, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Br and Pb were estimated in the tissueand blood samples. In the present study, concentration of Zn in the carcinoma gallbladder tissue is lessthan that of the normal gallbladder tissue. Tobacco habit could be one of the important factors todecrease the elemental concentrations in blood and tissue samples.

� 2009 Elsevier B.V. All rights reserved.

1. Introduction

Carcinoma of the gallbladder is the most common malignantdisease of the biliary tract. It is a highly fatal disease with dismalprognosis. It is the most common malignant lesion of the biliarytract and one of the most common among malignant neoplasmsof the digestive tract. There have been some attempts to under-stand the role played by trace elements in either initiating or pro-moting or inhibiting the growth of cancer [1–3]. In suchinvestigations, the concentrations of different elements in the tis-sues of the cancer-afflicted organ as well as the normal tissue ofthe same organ are measured employing a high precision tech-nique like external proton induced X-ray emission. Trace elementsare an important and emerging class of carcinogen [4]. Many traceelements are potent carcinogen in laboratory animals [5]. A few arepotent carcinogen and several are suspected human carcinogen [6].Trace elements are ubiquitous in both natural environment andwork place. Beyond this, trace elements are typically persistentwithin the natural and man made environment with growing con-centration in biosphere [7]. The trace elements absorbed by the

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+82 534229631.ay).

body enter the digestive system, pass through gastrointestinaltract and are deposited in the liver by blood stream. From the liverthey are carried to different organs for participating in biochemicalreactions. Usually, the ions of trace elements act as coordinationcentres to build up the structure of enzymes or proteins [8]. Thus,when the concentrations of the trace elements in the body differfrom the normal values, many clinical and pathological disordersarise. The excess or deficiency of these elements in the tissues ofthe cancer-afflicted organ when compared with that in the normalorgan, is sought to be correlated with the pathology of cancer ofthat organ.

Toxic elements like cadmium and nickel are promoters in thecancer process [4]. It was reported that the presence of specific ele-ments in human subjects is an indicator of cancer. The serum cop-per level and serum zinc level were reported to correlate withvarious cancers [9–12]. Zinc is a ubiquitous trace metal requiredfor the activity of over 300 metalloenzymes, including many in-volved in nucleic acid synthesis and cellular replication. Addition-ally, there are over 1400 zinc-finger proteins that participate in thegenetic expression of many proteins. Deficiency of zinc may resultin increased wound complications and cell-mediated immune dys-functions in humans and increased rates of tumor development inexperimental animals [13].

T.R. Rautray et al. / Nuclear Instruments and Methods in Physics Research B 267 (2009) 2878–2883 2879

2. Materials and methods

2.1. Sample preparation

The normal and carcinoma tissues of gallbladder of both sexeswere collected in the age group of 38 to 71 years. The tissue sam-ples were washed with deionised distilled water 5–6 times andfreeze dried in a lypholiser for 8 h till the samples got fully dried.After drying, the tissue samples were crushed to make pellets.For external PIXE and PIGE irradiation, 500 mg of powdered sam-ples were mixed with 500 mg of high pure cellulose powder in1:1 ratio by mass to make pellets of size 25 mm diameter in ahydraulic pressure and taken for analysis. Similarly, the blood sam-ples were pressed into pellets of size 13 mm diameter after lyphol-isation for vacuum PIXE irradiation.

2.2. External PIXE set-up

The external PIXE set-up at Institute of Physics (Fig. 1), which isthe unique of its kind in India, was installed by us in 2003 [14]. Theproton beam of 3 MeV energy was obtained from the 3 MV tandemtype horizontal pelletron accelerator (Model: 9SDH-2, make: Na-tional Electrostatics Corporation, Madison, USA) and collimatedby a graphite collimator to a beam size of 2 mm diameter. Thebeam was extracted into air using a KaptonTM foil (8 lm thick) atthe exit point of a vacuum scattering chamber [14]. The scatteringchamber has an inner diameter of 80 cm and was designed to caterto the requirements of the external beam as well as to serve for thecharged particle reaction studies for nuclear physics experiments.The beam was first focused and centered at the target location in-side the scattering chamber and then let through the thin Kaptonfoil placed at the exit port. The Kapton foil is used as exit windowdue to its several special characteristics like low beam-inducedbackground emission, minimal energy loss and resistance to radi-ation damage. The beam was allowed to travel a few cm in air afterwhich it irradiates the samples. Beam charge measurements werecarried out by using a rotating vane chopper designed by us [15].

For the measurements to be described below, the samples werekept in air over a sample stand (of 5 kg capacity) making an angleof 45� to the beam direction. The samples were irradiated withmaximum beam current of 30 nA. A Si (Li) detector (active area30 mm2) having energy resolution of 170 eV at 5.9 KeV placed at90� with respect to the beam direction was used to detect charac-teristic X-rays emitted from the target [14,16]. The detector has anentrance beryllium window of 8 lm thickness. A 25 lm thick alu-minium absorber (with 6% hole) was kept in front of the detector toattenuate the bremsstrahlung background and the dominant lowenergy X-ray peaks [17]. Spectra were recorded by using a PCbased multi channel analyser. The PIXE spectral analyses were per-

Fig. 1. External PIXE set-up at Institute of Physics.

formed using GUPIX-2000 software. This provides a non-linearleast square fitting of the spectrum, together with subsequent con-version of the fitted X-ray peak intensities into elemental concen-trations, utilizing the fundamental parameter method (FPM) forquantitative analysis. The PIGE spectral analyses were done usingthe ANGES (IAEA, Vienna) software [18–20].

3. Results and discussion

The external PIXE spectrum of a normal gallbladder tissue andcarcinoma gallbladder tissue are shown in Figs. 2 and 3 respec-tively. Similarly, the PIGE spectrum of a normal gallbladder tissueand carcinoma gallbladder tissue are shown in Figs. 4 and 5 respec-tively. The analysis of the tissue samples were carried out bysimultaneous external PIXE and PIGE technique whereas the anal-ysis of the blood samples of the healthy persons and the patientsaffected by gallbladder cancer were carried out by vacuum PIXEtechnique [21]. In the present study, seventeen trace elementsnamely Na, Mg, Al, K, Ca, Ti, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Brand Pb were estimated in both normal tissue and carcinoma tissue,normal blood and carcinoma blood samples of gallbladder whichare provided in Table 1. From PIGE technique, the concentrationof Na, Mg and Al have been established in the tissue samples. Inthe gallbladder carcinoma tissues, concentration of elementsnamely Na, Mg, Al, Ca, Zn and Se are less than that in the normalgallbladder tissues, whereas the concentration of K and Ca hasbeen decreased in case of the blood samples from the patients af-fected with gallbladder cancer. The concentration of elementsnamely Ti, Fe, Co, Ni and Cu have increased in the carcinoma tissuesamples. But average concentration of Cu has been increased incarcinoma blood samples.

The ten major elements in living bodies are H, C, N, O, Na, Mg, P,S, K and Ca. The total amount of these elements reaches 99% of allelements in the living bodies. The concentration of elements of N,Na, P, S and Ca are higher in animals, and concentration of ele-ments like O, Mg and K are lower than in plants. High contentsof these elements in animals are originating from proteins for Nand S, from bones for P and Ca, and from electrolytes for Na. Lowcontents of O, Mg and K in animals are due to lack of cellulosesas a major constituent to make cell wall membranes. Together withthese major elements, such minor elements as F, Si, Cr, Mn, Fe, Co,Ni. Cu, Zn, Se, Mo, Sn, I etc. are indispensable for animals. These ele-ments are distributed in each organ in a characteristic way. If the

Fig. 2. External PIXE spectrum of a normal gallbladder tissue.

Fig. 3. External PIXE spectrum of a carcinoma gallbladder tissue.

Fig. 4. PIGE spectrum of a normal gallbladder tissue.

Fig. 5. PIGE spectrum of a carcinoma gallbladder tissue.

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balance of these elements is broken, living bodies have to experi-ence ‘‘diseases” [22].

The concentration of Na in the carcinoma tissues is about 0.7times less than that in the normal tissues. But, the concentrationof Mg in the carcinoma gallbladder tissues is about 0.4 times lessthan that in the normal tissues. The relative concentration of ele-ments in normal versus carcinoma gallbladder tissue and bloodare depicted in Figs. 6 and 7 respectively. High intakes of dietaryfibre lower magnesium absorption. In contrast, high intakes of zincdecrease magnesium absorption and contribute to a shift towardnegative balance in adult males [23].

It was observed in the present work that K levels in the car-cinoma blood of gallbladder is lower than those in the normalblood samples. K is a major intracellular cation which is also ex-creted in gastrointestinal tract, saliva, gastric juice, bile, pancre-atic and intestinal juices. Prolonged hypokalemia (potassiumdeficiency) causes injury to myocardium and kidneys. The glyco-lytic enzyme ‘‘pyruvate kinase” requires potassium for its maxi-mum activity [24]. K deficiency causes acidosis, renal damageand cardiac arrest.

Shonk and his associates [25–27] studied the activities of differ-ent glycolytic enzymes in carcinoma of rectum and colon and inthe corresponding non-malignant counterparts. They observedthat glycolytic enzyme activities of the neoplastic tissues of rectumand colon are much higher than the activities in the correspondingnormal rectum and colon. They reported the ratio of neoplastic tonormal ‘‘pyruvate kinase” enzyme activity is 2.1 for colon and 2.4for rectum. Hence, it is expected that the levels of potassium whichmay be a cofactor for ‘‘pyruvate kinase” enzyme should be higherin the carcinoma tissue [28]. But the observation of low K levelsin the blood of carcinoma gallbladder in the present work doesnot support the above statement. This may possibly be due to dif-ferent organs.

Ca is a major constituent of bone and teeth. It is required forcoagulation of blood, regulation of neuromuscular irritability andmuscular contractility. Concentration of Ca in both the gallbladdercarcinoma tissue and blood samples is about 0.6 times less thanthat in the gallbladder normal tissue and blood. In a strictly oper-ational sense, Ca balance is determined by the relationship be-tween Ca intake and Ca absorption and excretion. A strikingfeature of the system is that relatively small changes in Ca absorp-tion and excretion can neutralise a high intake or compensate for alow one.

The concentration of Se in the carcinoma tissue samples isslightly less than that of the normal. Se has long been marked asa carcinogen. Nevertheless, the Committee on Medical and Biolog-ical effects of Environmental pollutants (1976) have reviewed theconfounding data and concluded that selenium may be an anti-car-cinogen. The inorganic Se at lower concentration acts as an anti-carcinogen and it is deficient in cancerous patients. Se can be agood metabolic risk modifier against cancer. Several elements likeNi, Se, Cu and Cr are considered essential at certain concentrations,but they are toxic when intake is excess [29]. The main source of Seis plant extracts and the cereals are the likely major dietarysources. The variation depends on the differences of Se contentof the soil of that particular place. Areas of high Se have higherprevalence of dental caries [30]. Selenium is a relatively toxic ele-ment. Intakes averaging 1.2 mg/day can induce changes in nailstructure. Chronic selenium intakes over 3.2 mg/day can result inthe loss of hair and nails, mottling of the teeth, lesions in the skinand nervous system, nausea, weakness and diarrhoea. Selenium,which is biologically important as an anion, is homeostatically reg-ulated by excretion, primarily in the urine but some also is ex-creted in the breath. Selenate, selenite and selenomethionine areall highly absorbed by the gastrointestinal tract; absorption per-centages for these forms of selenium are commonly found to be

Table 1Average concentration (in ppm by weight) of elements in normal gallbladder and carcinoma gallbladder tissue and blood.

Tissue Blood

Normal (n = 10) Carcinoma (n = 9) Normal (n = 10) Carcinoma (n = 9)

By PIGENa 717.2 ± 61.8 512.6 ± 47.2 – –Mg 8621.7 ± 551.2 3807.5 ± 227.5 – –Al 559.2 ± 50.4 262.3 ± 32.5 – –By external PIXEK 408.2 ± 38.9 359.4 ± 41.3 2082.7 ± 136.1 1834.4 ± 122.7Ca 616.3 ± 54.5 366.2 ± 40.8 321.7 ± 38.9 201.2 ± 31.2Ti 23.7 ± 3.8 37.2 ± 5.2 1.97 ± 0.4 2.01 ± 0.4Cr 11.1 ± 1.9 9.0 ± 1.7 1.07 ± 0.2 1.02 ± 0.2Mn 27.6 ± 3.6 23.4 ± 3.4 4.13 ± 0.8 3.21 ± 0.6Fe 1066.9 ± 93.8 1579.7 ± 113.4 597.3 ± 52.5 657.1 ± 48.7Co 8.25 ± 1.3 13.1 ± 2.3 0.74 ± 0.2 0.87 ± 0.2Ni 7.53 ± 1.3 12.3 ± 2.1 1.13 ± 0.3 1.74 ± 0.4Cu 11.1 ± 2.0 27.8 ± 4.3 1.10 ± 0.3 2.11 ± 0.4Zn 101.3 ± 16.9 75.6 ± 13.3 2.54 ± 0.5 2.06 ± 0.4As 3.8 ± 0.6 4.3 ± 0.7 0.11 ± 0.03 0.13 ± 0.03Se 3.14 ± 0.6 2.06 ± 0.4 0.16 ± 0.04 0.14 ± 0.04Br 12.6 ± 2.1 10.5 ± 1.8 1.04 ± 0.3 1.0 ± 0.3Pb 13.5 ± 2.1 11.8 ± 1.9 0.09 ± 0.03 0.16 ± 0.04

Fig. 6. Concentration of elements in normal versus carcinoma gallbladder tissue.

Fig. 7. Concentration of elements in gallbladder normal versus carcinoma blood.

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in the 80–90% range. Lead is very useful, but very harmful as wellwhen intake is more.

The concentration of Fe in carcinoma tissue samples havebeen increased as compared to the Fe level in normal tissue.But the concentration of Cu in the carcinoma blood and tissuesamples has increased as compared to the normal blood and tis-sue samples. Deficiency of Fe results in anaemia. It is responsiblefor oxygen transport and cellular respiration. Signs of Cu defi-ciency are anaemia, blood vessel necrosis, dyspigmentation, neu-ropathy, emphysema and diarrhoea. But excess intake of Cucauses stomach-ache, diarrhoea, vomiting, liver dysfunction andhemolysis.

Zinc contents in every organ, except for the cancer affected or-gan where Zn is enriched, are reported to become low if a human isattacked by cancer. This is caused by a high transfer rate of Zn towastes [22]. But, to contrary, in the present study, average concen-tration of Zn in the carcinoma gallbladder tissue is less than that ofthe normal gallbladder tissue.

Zinc is present in all body tissues and fluids. The total body zinccontent has been estimated to be 30 mmol (2 g). Plasma zinc has arapid turnover rate and it represents only about 0.l% of total bodyzinc content. This level appears to be under close homeostatic con-trol [31]. Deficiency of Zn causes poor growth, sexual infantilism inadolescents, loss of taste and delayed wound healing.

The utilisation of zinc depends on the overall composition of thediet. Experimental studies have identified a number of dietary fac-tors as potential promoters or antagonists of zinc absorption. Sol-uble low molecular weight organic substances, such as aminoand hydroxy acids facilitate zinc absorption. In contrast, organiccompounds forming stable and poorly soluble complexes with zinccan impair absorption. In addition, competitive interactions be-tween zinc and other ions with similar physicochemical propertiescan affect the uptake and intestinal absorption of zinc. The risk forcompetitive interactions seems mainly to be related to high dosesin the form of supplements or in aqueous solutions. However, atlevels present in food and at realistic fortification levels, zincabsorption appears not to be affected by iron and copper [32].The lack of specific and sensitive indexes for zinc status limitsthe possibilities for evaluating zinc requirements from epidemiol-ogic observations. Zinc requirements were estimated by using thefactorial technique (i.e. by adding the requirements for tissuegrowth, maintenance, metabolism and endogenous losses). Exper-imental zinc repletion studies with low zinc intakes have clearly

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shown that the body has a pronounced ability to adapt to differentlevels of zinc intakes by changing the endogenous zinc lossesthrough the kidneys, intestine and skin [33–37]. The concentrationof toxic elements like As and Pb did not show much variation be-tween the normal and carcinoma gallbladder tissue and blood.Moreover, As and Pb concentration may be attributed to be con-tamination during sample lypholisation, pelletization or handling.

The profound negative influence of malnutrition on cancertreatment is well recognised, with nutritional deficiencies impact-ing on all modalities of treatment. It therefore becomes critical toassess the nutritional status of patients before therapy and identifythose at greater risk for morbidity. Given the presence of carci-noma effects coupled with years of tobacco and alcohol abuse,the high incidence of nutritional deficiency in the gallbladder isnot surprising.

Diet may influence human carcinogenesis in six general ways[38]: (i) Diet provides the carcinogens or their immediate precur-sors, (ii) Diet facilitates or inhibits the endogenous production ofcarcinogens, (iii) The modification of carcinogens by metabolicactivation or inactivation could be affected by other dietary com-ponents, (iv) Increasing or impeding the delivery of carcinogensto their site of action may be influenced by dietary changes, (v)Diet may alter the susceptibility of tissues to cancer induction orgrowth by dietary effects or tissue metabolism, (vi) Diet may alterthe body’s capacity to eliminate transformed cells.

In the field of nutrition and cancer, turmeric is emphasised as ananti-cancer agent. It prevents the formation of preneoplastic le-sions induced in the initiation as well as post initiative phase.The As concentrations are found relatively high in spices, whereaslead and iron are found more in tobacco-products. Clinical studieshave shown that iron promotes cancer cell growth. It acts more likea co-carcinogen than a primary one and in case of tobacco user’slocal absorption of concentrated iron in oral region could be high.Tobacco addicts who normally require more antioxidants as com-pared to a normal person tend to consume a less adequate dietwith subnormal antioxidant levels than do others [39].

From the epidemiological evidences and different animal mod-els, it is established that Mn and Se are non-carcinogens. But, ex-cess exposure to Cr, As, Cd, Ni, Be, Co, Pb, Si and Sn may lead tocancer. The carcinogenicity of Al, Cu, Fe, Mo, Sc, Ti, V and Zn areyet to be established [40]. Daily nutritional intake of elements ofdifferent individuals from different areas and studies on trace ele-ment imbalances in different environment and different biomedi-

Table 2Concentration (in ppm by weight) of some tobacco-products by PIXE analysis.

K% Ca% Mn Fe C

Cigarette (n = 8) 0.63 ± 0.04 2.03 ± 0.08 151.6 ± 23.6 687.6 ± 68.7 1Bidi (n = 8) 0.76 ± 0.06 4.21 ± 0.04 97.8 ± 19.7 1397.2 ± 159.5 1Gutkha (n = 8) 1.03 ± 0.06 2.01 ± 0.07 73.6 ± 17.8 472.8 ± 72.4 9Zarda (n = 8) 0.69 ± 0.05 1.76 ± 0.07 49.5 ± 10.3 484.9 ± 83.7 1Khaini (n = 7) 0.87 ± 0.04 4.01 ± 0.04 157.4 ± 28.5 615.8 ± 117.3 2Gudakhu (n = 7) 0.81 ± 0.04 1.63 ± 0.07 121.4 ± 17.0 992.3 ± 147.3 1

Table 3Average concentration (in lg/ml) of various elements of human blood with respect to tob

Groups Average elemental concentrations of whole blood

I (No habit) II (Light habits)

K 2132.5 ± 127.3 1982.6 ± 120.4Ca 272.4 ± 45.2 263.4 ± 46.4Fe 532.7 ± 82.6 504.0 ± 80.6Cu 1.06 ± 0.22 0.91 ± 0.20Zn 2.41 ± 0.50 2.32 ± 0.49Se 0.16 ± 0.03 0.13 ± 0.02Pb 0.07 ± 0.01 0.07 ± 0.01

cal samples may lead to better diagnosis of diseases like cancerwhich will definitely guide the clinicians to supplement or with-draw the elements and minerals to the human body for cure andmaintenance of better health.

It was observed from Table 2 that all the tobacco-products werevery highly enriched with multi-elements. The present results fortobacco-shoots when compared with those reported by Oliveiraet al. [41], there is an exception in the strontium and copper levels.Low Sr-levels and high Cu-levels were observed in the tobacco-samples. These elemental variations in the tobacco-chemistrycould be due to some agricultural and industrial factors like pre-harvest and post-harvest operations, pre-blend operations, drying,storing, processing, flavor addition and casings.

Among the common smoking tobaccos, it was found that the In-dian bidi, which contains raw tobacco leaves, have higher elemen-tal levels than that of the cigarettes. Clinical studies have shownthat Fe promotes cancer cell growth and it acts more like a co-car-cinogen [42]. All the tobacco-products were found to be veryhighly enriched with iron. The branded tobacco-products like gut-kha and zarda, whose other ingredients are betel nuts, lime, carda-mom catechu, silver leaves, aromatic spices and saffron, havecomparatively lower iron level than that of the raw tobacco-prod-ucts such as gudakhu. Exposure to arsenic is associated with the onset of cancers of skin and lung.

Table 3 shows the elemental variations in human blood with re-spect to their tobacco habits. From the results, it was observed thatall the elemental levels in human blood decrease with respect toincrease in the tobacco habits. The selenium and copper contentsin the blood decreases significantly in case of heavy users of tobac-co in comparison with that of the control group, who have no to-bacco habits [43].

The blood iron and copper values in case of moderate and heavyusers of tobacco has become lower than the reported normalranges [44]. Of course, these lower elemental values could bedue to the inadequate and poor dietary habits of the tobacco users[45]. The copper and zinc ratio, which is an important index in dis-ease activity [46], decreases significantly with respect to increasinghabits of tobacco.

4. Conclusion

Decreasing levels of Zn and Se in the tissue samples have a def-inite importance in the oxidative processes in the human body.

u Zn As Se Sr Pb

5.4 ± 3.1 29.6 ± 5.7 0.17 ± 0.03 0.10 ± 0.02 14.8 ± 3.1 0.47 ± 0.102.5 ± 2.0 61.8 ± 9.3 0.31 ± 0.05 0.15 ± 0.03 19.6 ± 4.0 1.25 ± 0.22.6 ± 1.6 71.1 ± 12.4 0.23 ± 0.05 0.14 ± 0.03 15.3 ± 2.9 0.68 ± 0.163.6 ± 2.7 43.4 ± 12.7 0.20 ± 0.04 0.11 ± 0.02 21.5 ± 3.8 0.71 ± 0.150.3 ± 3.1 18.2 ± 3.5 0.19 ± 0.04 0.15 ± 0.03 19.3 ± 4.1 0.69 ± 0.144.8 ± 2.2 72.5 ± 16.2 0.30 ± 0.06 0.19 ± 0.04 15.8 ± 2.8 0.97 ± 0.18

acco habits by PIXE technique.

III (Moderate habits) IV (Heavy habits)

1971.8 ± 121.0 1711.3 ± 198.2251.1 ± 44.9 239.3 ± 45.2372.8 ± 69.8 359.3 ± 71.80.83 ± 0.17 0.61 ± 0.172.21 ± 0.46 1.68 ± 0.470.10 ± 0.02 0.07 ± 0.020.07 ± 0.01 0.06 ± 0.01

T.R. Rautray et al. / Nuclear Instruments and Methods in Physics Research B 267 (2009) 2878–2883 2883

But, to contrary, in the present study, concentration of Zn in thecarcinoma gallbladder tissue is less than that of the normal gall-bladder tissue. Tobacco habit could be one of the important factorsto decrease the elemental concentrations in blood and tissue sam-ples. Gradual increase in tobacco habits decreases the dietary in-take of a person and hence trace element levels in blood.Therefore, proportionately adequate supplementation of trace ele-ments to tobacco users is very much essential to scavenge the freeradicals. The local absorption of highly concentrated iron and cal-cium from the tobacco-products could be dangerous. Tobaccoproducts are rich in multi-elements. The more the local absorptionof multi-elements, the more is the local irritation and subsequentdevelopment of cancer.

Acknowledgements

The help extended by the staff members of Institute of Physics(IOP) is acknowledged and the authors would like to thank theauthorities of IOP for providing beam time to carry out the currentresearch.

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