Research Article A Novel Oxidative Stress Mediator in ...downloads.hindawi.com/journals/mi/2016/6761050.pdf · Conclusion . is is the rst study examining the thiol/disulphide homeostasis

Research ArticleA Novel Oxidative Stress Mediator in Acute AppendicitisThiolDisulphide Homeostasis

Sefa Ozyazici1 Faruk Karateke1 Umit Turan1 Adnan Kuvvetli1 Huseyin Kilavuz1

Burak Karakaya1 PJnar Ozaltun2 Murat AlJsJk3 and Ozcan Erel3

1Department of General Surgery Adana Numune Training and Research Hospital 01170 Adana Turkey21209 Sok Akay Apt No 6 Palmiye Mahallesi Mersin Turkey3Department of Biochemistry Faculty of Medicine Yildirim Beyazit University 01170 Ankara Turkey

Correspondence should be addressed to Faruk Karateke karatekefarukhotmailcom

Received 12 April 2016 Revised 6 July 2016 Accepted 19 July 2016

Academic Editor Julio Galvez

Copyright copy 2016 Sefa Ozyazici et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Aim To investigate the role of a novel oxidative stress marker thioldisulphide homeostasis in patients diagnosed with acuteappendicitis (AA) Methods In this study seventy-one (43 male and 28 female) patients diagnosed with AA and 71 (30 male and41 female) healthy volunteers were included Age gender body mass index (BMI) haemoglobin (Hb) white blood cell (WBC)c-reactive protein (CRP) and thioldisulphide homeostasis parameters (native thiol total thiol disulphide disulphidenative thiolnative thioltotal thiol and disulphidetotal thiol ratios) were compared between the groups Thioldisulphide homeostasis wasdetermined by a newly developedmethod by Erel andNeselioglu ResultsThe native thiol total thiol and the native thioltotal thiolratio levels were statistically significantly decreased in theAAcomparedwith the control group (119901 lt 0001) Disulphide level and theratios of disulphidenative thiol and disulphidetotal thiol were higher in the AA group than in the control group (119901 lt 0001)Therewas a negative correlation of CRPwith native thiol total thiol and native thioltotal thiol ratio while there was a positive correlationof CRP with disulphidenative thiol and disulphidetotal thiol in the AA group In the stepwise regression model risk factors asdisulphidenative thiol (OR = 1368 119901 = 0018) and CRP (OR = 1635 119901 = 0003) were determined as predictors of perforatedappendicitis compared to the nonperforated group Conclusion This is the first study examining the thioldisulphide homeostasisas a diagnostic aid in AA and establishing thioldisulphide homeostatis balance shifted towards the disulphide formation due tothiol oxidation Further studies are needed to optimize the use of this novel oxidative stress marker in AA

1 Introduction

Acute appendicitis (AA) is the most common abdominalemergency worldwide that requires surgical intervention inthe daily clinical practice of general surgery [1]The diagnosisof acute appendicitis has been primarily made with clinicalsymptoms and physical examination Since symptoms of AAoverlap with other abdominal and gynecologic conditionsachieving an accurate diagnosis is still a clinical challenge forsurgeons and there is a lack of adequate specific biomarkersfor AA To support the diagnosis preoperative scoringsystems such as the Alvarado score [2] were developedhowever these systems are rarely used In clinical practicethe most commonly used laboratory tests for the diagnosisof AA are WBC and CRP The nonspecific presentation and

limited diagnostic accuracy of laboratory results often leadto an inadequate diagnosis necessitating radiological imag-ing of the abdomen Ultrasonography (US) and computedtomography (CT) scanning can enhance diagnostic accuracyin patients with suspected AA [3]

Although the pathophysiology of AA has been describedin detail the factors involving the progression of AA arestill being investigated Recently it was found that one ofthese factors of interest is oxidative stress The possiblerole of oxidative stress parameters in the progression ofAA has been demonstrated in a few studies [4ndash6] Somebiochemical markers such as those used to identify oxidativestress and inflammation have been proposed as indicatorsof the presence and extent of acute appendicitis [5 7] Ithas been known that plasma thiols are physiological free

Hindawi Publishing CorporationMediators of InflammationVolume 2016 Article ID 6761050 6 pageshttpdxdoiorg10115520166761050

2 Mediators of Inflammation

radical scavengers and they may serve as an antioxidantfunction by several mechanisms It has been known thatthe measurement of plasma total thiol and determiningthioldisulphide homeostasis are a good reflection of excessfree radical generation in several diseases [8ndash10]

In this study we hypothesized that oxidative stress playsa role in the pathogenesis of AA and we aimed to investigatea novel oxidative stress marker thioldisulphide homeostasisin patients diagnosed with AA as well as its correlation withother inflammatory markers such as CRP and WBC

2 Materials and Methods

21 Study Population This study was carried out in AdanaNumune Training and Research Hospital General SurgeryClinic between April 2015 and December 2015 A total of142 participants consisting of 71 patients who underwentoperations with diagnosis of AA over the age of 18 years and71 healthy volunteers for the control group were includedin the study The control group consisted of healthy patientswith similar demographic characteristics to the patient groupand who have applied to our hospital with dyspeptic com-plaints but without a known chronic disease smokers andalcohol users or drug users Participants were also excludedfrom the study if they were pregnant or had a history ofmalignancy or abdominal traumawithin 7 days of admissionParticipants in the control group were included according totheir reference sequence The diagnosis of acute appendicitiswas made with clinical symptoms and physical examinationas well as laboratory tests and imaging techniques such asultrasonography and computed tomography This study hasbeen designed in accordance with 2013 Brazil version ofHelsinki Declaration and was approved by the local EthicsCommittee All participants have provided written consentThe principles of good clinical practices were followed duringthe study period

22 Biochemical Parameters Venous blood samples weretaken to measure thioldisulphide homeostasis parametersof all participants who were included in the study In AApatients blood samples were taken when they admitted toour clinic before surgical intervention In healthy volunteersblood samples were taken when they were admitted to ourpoliclinic After blood samples were quickly centrifuged at1500 rpm for ten minutes plasma and serum samples wereseparated Serum samples have been stored at minus80∘C untilall samples were obtained Then the plasma samples testedfor thioldisulphide levels were shipped to biochemical lab-oratory of Ataturk Training and Research Hospital AnkaraTurkey after all samples were collected Hemogram bio-chemistry and CRP levels of all participants were measuredat the time they were enrolled in the study

Thioldisulphide levels were analyzed with a newly devel-oped method by Erel and Neselioglu [11] In summaryreducible disulphide bonds were first reduced to form freefunctional thiol groups Unused reductant sodium borohy-dride was consumed and removedwith formaldehyde and allthiol groups including reduced and native ones were detected

after reaction with 551015840-dithiobis-(2-nitrobenzoic) acid Halfof the difference between total and native thiols provided thedynamic disulphide amount (-S-S) After the determinationof native thiol (-SH) and disulphide (-S-S) amount nativethioldisulphide ratio (-S-S--SH) was calculated Laboratorystaff performing the plasma thioldisulphide homeostasismeasurement analysis were blinded to the patientsrsquo clinicalinformation and outcome and results were not available tothe treating physicians study staff or investigators duringstudy period

23 Statistical Analysis Statistical Package for Social Sciences(SPSS) for Windows 20 (IBM SPSS Inc Chicago USA)program was used for statistical analyses Kolmogorov-Smirnov test was used to determine the distribution of dataContinuous variables with normal distribution were given asmean plusmn standard deviation and continuous variables withoutnormal distribution were given as median interquartile range[IQR] Categorical variables were expressed as numbersand percentage Continuous variables were compared withindependent sample 119905-test ANOVA Mann Whitney 119880 testand Kruskal-Wallis 119867 test where appropriate Chi-squaretest and Fisherrsquos exact chi-square test were used to comparecategorical variables The relationship between the numericparameters was analyzed by Pearson and Spearman cor-relation analysis Stepwise multivariable logistic regressionanalysis was used to identify independent predictors of AArisk A 119901 lt 005 was considered significant for statisticalanalyses

3 Results

A total of 142 individuals were evaluated in the study Nosignificant difference was observed between the AA andcontrol groups with respect to age Hb and BMI (119901 gt005) The native thiol total thiol and the native thioltotalthiol ratio levels were statistically significantly decreased(119901 lt 0001) in the patients with AA compared with thecontrol subjects On the other hand disulphide level andthe ratios of disulphidenative thiol and disulphidetotal thiolwere higher in the AA group than in the control group(119901 lt 0001) The demographics and the thioldisulfidehomeostasis parameters of the patients and healthy controlsare summarized in Table 1

In the appendectomy group patients were further dividedinto two categories nonperforated appendicitis (119899 = 54) andperforated appendicitis (119899 = 17) There was no significantdifference between the two groups in terms of sex BMI andHb (119901 gt 005) The levels of native thiol and total thiol(119901 = 0003 and 119901 = 0005 resp) as well as the nativethioltotal thiol ratio (119901 = 004) were lower in the perforatedgroup when compared to those of the nonperforated groupOn the other hand disulphide level and the ratios of disul-phidenative thiol and disulphidetotal thiol were higher inthe perforated group than in the nonperforated group (119901 =0027 119901 = 0001 and 119901 = 0001 resp) The demographicsand the thioldisulfide homeostasis parameters of the patientsin AA group are summarized in Table 2

Mediators of Inflammation 3

Table 1 The demographics and laboratory findings of AA and control groups

Variables Acute appendicitis Control119901 value

(119899 = 71) (119899 = 71)Gender 119899 ()

Male 43 (606) 30 (423) 004Female 28 (394) 41 (577)

Age (years) 324 plusmn 91 301 plusmn 89 NSBMI (kgm2) 217 plusmn 29 238 plusmn 31 NSWBC (120583L) 135 plusmn 27 71 plusmn 15 lt0001CRP (mgL) 13 (18) 06 (3) lt0001Hemoglobin (gdL) 136 plusmn 09 136 plusmn 11 NSNative thiol (120583molL) 2707 plusmn 681 3865 plusmn 585 lt0001Total thiol (120583molL) 3006 plusmn 668 4069 plusmn 602 lt0001Disulphide (120583molL) 186 plusmn 45 105 plusmn 33 lt0001Disulphidenative thiol () 73 plusmn 31 27 plusmn 09 lt0001Disulphidetotal thiol () 61 plusmn 22 25 plusmn 08 lt0001Native thioltotal thiol () 872 plusmn 51 949 plusmn 16 lt0001119901 lt 005 statistical significanceParameters were expressed as mean plusmn SD and median (interquartile range (IQR))BMI body mass index (kgm2) WBC white blood cell CRP C-reactive protein

Table 2 The demographics and laboratory findings of patients in AA group

Variables Perforated appendicitis Nonperforated appendicitis119901 value

(119899 = 17) (119899 = 54)Gender 119899 ()

Male 13 (765) 30 (556) NSFemale 4 (235) 24 (444)

Age (years) 365 plusmn 85 311 plusmn 89 0031BMI (kgm2) 225 plusmn 34 214 plusmn 27 NSWBC (120583L) 146 plusmn 33 131 plusmn 24 0044CRP (mgL) 78 (17) 12 (106) lt0001Hemoglobin (gdL) 139 plusmn 11 136 plusmn 09 NSNative thiol (120583molL) 2283 plusmn 602 2841 plusmn 653 0003Total thiol (120583molL) 2612 plusmn 683 3127 plusmn 619 0005Disulphide (120583molL) 215 plusmn 64 177 plusmn 33 0027Disulphidenative thiol () 96 plusmn 32 66 plusmn 26 lt0001Disulphidetotal thiol () 79 plusmn 22 56 plusmn 19 lt0001Native thioltotal thiol () 841 plusmn 44 881 plusmn 49 0004119901 lt 005 statistical significanceParameters were expressed as mean plusmn SD and median (interquartile range (IQR))BMI body mass index (kgm2) WBC white blood cell CRP C-reactive protein

Table 3 shows the correlation analysis between thioldisulphide homeostasis parameters and other characteristicfeatures with study population We determined a negativecorrelation of CRP with native thiol total thiol and nativethioltotal thiol ratio while we determined a positive correla-tion of CRP with disulphidenative thiol and disulphidetotalthiol in the AA groupThere was a negative correlation of agewith total thiol and native thioltotal thiol in the AA groupThere was a negative correlation of age with native thiol andtotal thiol in the control group

In the stepwise regression model formed with risk factorssuch as age sex BMIWBC Hb CRP native thiol total thioldisulphide disulphidenative thiol disulphidetotal thioland native thioltotal thiol risk factors as disulphidenativethiol (OR = 1368 119901 = 0018) and CRP (OR = 1635119901 = 0003) were determined as predictors of perforatedappendicitis compared to the nonperforated group It wasdetermined that 1-unit increase in disulphidenative thiol ratepredicts 1368 times the risk of having perforated appendicitisand 1-unit increase in CRP level predicts 1635 times the risk


Table3Correlatio

nanalysisbetweencharacteris

ticsa

ndlabo

ratory

finding

swith

study

popu

latio

n

Varia

bles

Nativethiol

Disu

lphide

Totalthiol

Disu

lphidenativethiol

Disu

lphideto

talthiol

Nativethiolto

talthiol

119903119901

119903119901

119903119901

119903119901

119903119901

119903119901

Groups

Con

trol

Age

(years)

minus0384

0001lowastminus0077

0522minus0378

0001lowast

0125

0297

0131

0277

minus0125

0299

BMI(kgm2

)minus0020

0867minus0106

0378minus0039

0746minus0133

0268

minus0134

0264

0138

0250

WBC

(120583L)

0166

0166

0194

0104

0174

0147

0033

0785

0038

0754

minus0019

0876

CRP(m

gL)

0085

0479

0080

0505

0084

0484minus0016

0893

0003

0979

0035

0774

Hem

oglobin(gdL)

0169

0159minus0057

0639

0164

0171minus0078

0520

minus0076

0529

0079

0512

AA Age

(years)

minus0161

0181

0101

0401minus0268

0024lowast

0192

0109

0179

0135

minus0292

0013lowast

BMI(kgm2

)minus0224

0061

0119

0323minus0154

0201

0128

0286

0136

0260

minus0230

0053

WBC

(120583L)

minus0212

0075minus0017

0888minus0114

0345minus0022

0854

minus0003

0977

0079

0512

CRP(m

gL)

minus0322

000

6lowast0114

0344minus0377

0001lowast

0302

0011lowast

0313

0008lowast

minus0234

0050lowast

Hem

oglobin(gdL)

0094

0434

0139

0247

0120

0320minus0084

0484

minus0089

0461

0041

0733

lowast

119901lt005statisticalsig

nificance

BMIbo

dymassind

ex(kgm2

)WBC

whitebloo

dcellCR

PC-

reactiv

eprotein


Table 4 Regression analysis and independent risk factors predicting perforated appendicitisCRP C-reactive protein

Variables OR 95 CI119901 valuelowast

Lower UpperDisulphidenative thiol 1368 1056 1773 0018CRP 1635 1183 2260 0003Nagelkerke 1198772 = 0586 119901 lt 0001lowastAge sex BMI and laboratory findings are included in the stepwise logistic regression modellowast

119901 lt 005 statistical significance

of having perforated appendicitisThese data are summarizedin Table 4

4 Discussion

Acute appendicitis is still a clinical challenge for physiciansas there is no single test that reliably differentiates AAfrom other causes of acute abdomen Developing specificdiagnostic tests for AA would both prevent unnecessarysurgery and help to define the advanced stages of thedisease requiring urgent intervention such as perforatedappendicitis which is associated with increased morbidityandmortality Several plasmamarkers have been investigatedin order to improve the diagnosis of AA [12] In addition toinflammatorymarkers oxidative stressmarkers have recentlybecome an area of interest for investigators In the literatureexperimental and clinical studies addressing the associationbetween AA and oxidative stress are limited [5 13 14] Ithas been reported that the serum levels of oxidative stressmarkers change in patients with AA determining that theimbalance between oxidant and antioxidant defense systemsmay play a role in the pathogenesis of AA which couldbe either etiologic or a result of inflammation [9 13 15]Therefore these serum markers may be used to aid thediagnosis of AA and to determine the extent of the disease

As is known thiols are a class of organic compounds thatcontain a sulfhydryl group (-SH) which is composed of ahydrogen and a sulphur atom attached to a carbon atom [16]Those disulphide bonds can be reduced back to thiol groupstherefore thioldisulphide homeostasis is maintained [17]Thiols contribute the major portion of the total antioxidantspresent in the body and play an important role in defenseagainst reactive oxygen species [18] and also play criticalroles in programmed cell death detoxification antioxidantprotection and regulation of cellular enzymatic activity [1119] Recently it is known that an abnormal thioldisulfidehomeostasis state is involved in the pathogenesis of variousacute and chronic diseases [10 11] Measuring thiols in serumprovides an indirect reflection of the antioxidative defense[10 11 18ndash20]Themeasurement of dynamic thioldisulphidefirst started by a new automated method developed by Ereland Neselioglu [11] This study aimed to determine the statusof dynamic thioldisulphide homeostasis by this newmethodin AA Furthermore we investigated the correlation betweenthioldisulphide homeostasis parameters and other clinicalfeatures in AA patients

Previously Yilmaz et al [9] and Dumlu et al [13] founddecrease in thiol levels in appendicitis patients compared

with controls Under oxidative stress disulphide level isexpected to increase as thiol level decreases However thistopic was not studied before in patients with AA To thebest of our knowledge this is the first study that investigatedthioldisulphide homeostasis as a novel marker of oxidativestress in patients with AA and compared the results withhealthy controls In our study we demonstrated that thelevels of native thiol and total thiol and the native thioltotalthiol ratio are lower in patients with AA as compared tohealthy individuals Besides we also demonstrated for thefirst time that disulphide level and disulphidenative thioland disulphidetotal thiol ratios all of which are formed asa result of thiol oxidation are higher in patients with AAthan in the healthy controls In other words thioldisulphidehomeostasis was found to shift towards disulphide formationThioldisulphide homeostasis parameters were similar as forthe subgroup analysis of AA patients We assume that thissituation shows the higher level of oxidative stress in perfo-rated appendicitis patients compared to that of nonperforatedappendicitis

In clinical practice CRP and WBC values are used topredict the severity of inflammation in AA Previous studieshave shown that the greater the degree of appendicularinflammation the greater the CRP value reachingmaximumvalues in cases of perforation [5 7] Similarly our studyhas disclosed a significant increase in CRP in appendicitisprogressed to perforation Furthermore correlation analysisshowed a negative correlation of CRP with native thioltotal thiol and native thioltotal thiol ratio while a positivecorrelation of CRP with disulphidenative thiol and disul-phidetotal thiol was noted In the stepwise logistic regressionanalysis formed with risk factors it was determined thatdisulphidenative thiol and CRP are independent predictorsof perforated appendicitis Therefore with the results of thisstudy we can say that when the increase of disulphidenativethiol ratio due to severity of inflammation and its positivecorrelation are considered together it is possible to concludethat disulphidenative thiol ratio could be related with pro-gression and used as a marker of disease activity Althoughplasma thioldisulphide homeostasis parameters cannot beused in the diagnosis of AA a shifting towards disulphide inits value by time might be considered as a predictor of theprogression of inflammation to the perforation in AA cases

There are several limitations in this study that shouldbe taken into consideration First this was a pilot studyrepresenting an initial investigation into the relationshipbetween AA and thioldisulphide homeostasis parametersSecond is inclusion of relatively small number of patients


who were admitted to a single center Other diagnosticaids such as procalcitonin and imaging techniques such asultrasonography andor computed tomography andAlvaradoscore have not been correlated with thioldisulphide home-ostasis parameters Lastly the plasma samples tested for thethioldisulphide homeostasis were frozen and shipped to asingle laboratory remote from our hospital rather than on-site using fresh plasma samples as would be done in clinicalpractice Further longitudinal studies on a larger patientpopulation are needed to determine whether alterations inthioldisulphide homeostasis could be predictive risk factorsfor AA

5 Conclusion

This study demonstrated that dynamic thioldisulphidehomeostasis shifted towards disulphide formation as a resultof thiol oxidation in patients with AA Prospective andrandomized controlled trials are necessary to confirm thepathophysiologic role of thioldisulphide homeostasis in AAFurther studies are required to optimize the use of this noveloxidative stress marker in conjunction with other establishedapproaches

Competing Interests

The authors have no competing interests to declare

Authorsrsquo Contributions

Sefa Ozyazici was responsible for conception and design andwriting the paper FarukKaratekewas responsible for analysisand interpretation of data and final approval of the version tobe published Umit Turan Huseyin Kilavuz Burak Karakayaand Adnan Kuvvetli were responsible for involving the datacollection Pınar Ozaltun handled statistical analysis MuratAlısık was responsible for laboratory analysis Ozcan Erel wasresponsible for revising it critically

References

[1] D J Humes and J Simpson ldquoAcute appendicitisrdquo The BritishMedical Journal vol 333 no 7567 pp 530ndash534 2006

[2] A Alvarado ldquoA practical score for the early diagnosis of acuteappendicitisrdquo Annals of Emergency Medicine vol 15 no 5 pp557ndash564 1986

[3] A Van Randen W Lameris H W Van Es et al ldquoA comparisonof the accuracy of ultrasound and computed tomography incommon diagnoses causing acute abdominal painrdquo EuropeanRadiology vol 21 no 7 pp 1535ndash1545 2011

[4] U Koltuksuz E Uz S Ozen M Aydinc A Karaman andO Akyol ldquoPlasma superoxide dismutase activity and malondi-aldehyde level correlate with the extent of acute appendicitisrdquoPediatric Surgery International vol 16 no 8 pp 559ndash561 2000

[5] M Ozdogan A O Devay A Gurer et al ldquoPlasma total anti-oxidant capacity correlates inversely with the extent of acuteappendicitis a case control studyrdquo World Journal of EmergencySurgery vol 1 no 1 article 6 2006

[6] A Satomi T Hashimoto SMurakami et al ldquoTissue superoxidedismutase (SOD) activity and immunohistochemical stainingin acute appendicitis correlation with degree of inflammationrdquoJournal of Gastroenterology vol 31 no 5 pp 639ndash645 1996

[7] D RMcGowanHM Sims K ZiaMUheba and I A ShaikhldquoThe value of biochemical markers in predicting a perforationin acute appendicitisrdquo ANZ Journal of Surgery vol 83 no 1-2pp 79ndash83 2013

[8] M-L Hu ldquoMeasurement of protein thiol groups and glu-tathione in plasmardquoMethods in Enzymology vol 233 pp 380ndash385 1994

[9] F M Yilmaz G Yilmaz M F Erol S Koklu and D YucelldquoNitric oxide lipid peroxidation and total thiol levels in acuteappendicitisrdquo Journal of Clinical Laboratory Analysis vol 24 no2 pp 63ndash66 2010

[10] M Yuksel I Ates M Kaplan et al ldquoThe dynamicthioldisulphide homeostasis in inflammatory bowel diseaseand its relation with disease activity and pathogenesisrdquoInternational Journal of Colorectal Disease vol 31 no 6 pp1229ndash1231 2016

[11] O Erel and S Neselioglu ldquoA novel and automated assay forthioldisulphide homeostasisrdquo Clinical Biochemistry vol 47 no18 pp 326ndash332 2014

[12] D H S M Schellekens K W E Hulsewe B A C Van Ackeret al ldquoEvaluation of the diagnostic accuracy of plasma markersfor early diagnosis in patients suspected for acute appendicitisrdquoAcademic Emergency Medicine vol 20 no 7 pp 703ndash710 2013

[13] E G Dumlu M Tokac B Bozkurt et al ldquoCorrelation betweenthe serum and tissue levels of oxidative stress markers and theextent of inflammation in acute appendicitisrdquo Clinics vol 69no 10 pp 677ndash682 2014

[14] M Kaya M E Boleken T Kanmaz O Erel and S YucesanldquoTotal antioxidant capacity in children with acute appendicitisrdquoEuropean Journal of Pediatric Surgery vol 16 no 1 pp 34ndash382006

[15] T V Zhavoronok E A Stepovaia N V Riazantseva et alldquoImpaired oxidative metabolism in acute inflammatory dis-easesrdquo Klinicheskaia Laboratornaia Diagnostika no 12 pp 10ndash14 2006

[16] C K Sen and L Packer ldquoThiol homeostasis and supplementsin physical exerciserdquoTheAmerican Journal of Clinical Nutritionvol 72 supplement 2 pp 653Sndash669S 2000

[17] D P Jones and Y Liang ldquoMeasuring the poise of thioldisulfidecouples in vivordquo Free Radical Biology and Medicine vol 47 no10 pp 1329ndash1338 2009

[18] Y R Chianeh and K Prabhu ldquoProtein thiols as an indicator ofoxidative stressrdquo Archives Medical Review Journal vol 23 pp443ndash456 2014

[19] M L Circu and T Y Aw ldquoReactive oxygen species cellu-lar redox systems and apoptosisrdquo Free Radical Biology andMedicine vol 48 no 6 pp 749ndash762 2010

[20] N Dirican A Dirican O Sen et al ldquoThioldisulfide home-ostasis a prognostic biomarker for patients with advanced non-small cell lung cancerrdquo Redox Report vol 21 no 5 pp 197ndash2032016

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radical scavengers and they may serve as an antioxidantfunction by several mechanisms It has been known thatthe measurement of plasma total thiol and determiningthioldisulphide homeostasis are a good reflection of excessfree radical generation in several diseases [8ndash10]

In this study we hypothesized that oxidative stress playsa role in the pathogenesis of AA and we aimed to investigatea novel oxidative stress marker thioldisulphide homeostasisin patients diagnosed with AA as well as its correlation withother inflammatory markers such as CRP and WBC

2 Materials and Methods

21 Study Population This study was carried out in AdanaNumune Training and Research Hospital General SurgeryClinic between April 2015 and December 2015 A total of142 participants consisting of 71 patients who underwentoperations with diagnosis of AA over the age of 18 years and71 healthy volunteers for the control group were includedin the study The control group consisted of healthy patientswith similar demographic characteristics to the patient groupand who have applied to our hospital with dyspeptic com-plaints but without a known chronic disease smokers andalcohol users or drug users Participants were also excludedfrom the study if they were pregnant or had a history ofmalignancy or abdominal traumawithin 7 days of admissionParticipants in the control group were included according totheir reference sequence The diagnosis of acute appendicitiswas made with clinical symptoms and physical examinationas well as laboratory tests and imaging techniques such asultrasonography and computed tomography This study hasbeen designed in accordance with 2013 Brazil version ofHelsinki Declaration and was approved by the local EthicsCommittee All participants have provided written consentThe principles of good clinical practices were followed duringthe study period

22 Biochemical Parameters Venous blood samples weretaken to measure thioldisulphide homeostasis parametersof all participants who were included in the study In AApatients blood samples were taken when they admitted toour clinic before surgical intervention In healthy volunteersblood samples were taken when they were admitted to ourpoliclinic After blood samples were quickly centrifuged at1500 rpm for ten minutes plasma and serum samples wereseparated Serum samples have been stored at minus80∘C untilall samples were obtained Then the plasma samples testedfor thioldisulphide levels were shipped to biochemical lab-oratory of Ataturk Training and Research Hospital AnkaraTurkey after all samples were collected Hemogram bio-chemistry and CRP levels of all participants were measuredat the time they were enrolled in the study

Thioldisulphide levels were analyzed with a newly devel-oped method by Erel and Neselioglu [11] In summaryreducible disulphide bonds were first reduced to form freefunctional thiol groups Unused reductant sodium borohy-dride was consumed and removedwith formaldehyde and allthiol groups including reduced and native ones were detected

after reaction with 551015840-dithiobis-(2-nitrobenzoic) acid Halfof the difference between total and native thiols provided thedynamic disulphide amount (-S-S) After the determinationof native thiol (-SH) and disulphide (-S-S) amount nativethioldisulphide ratio (-S-S--SH) was calculated Laboratorystaff performing the plasma thioldisulphide homeostasismeasurement analysis were blinded to the patientsrsquo clinicalinformation and outcome and results were not available tothe treating physicians study staff or investigators duringstudy period

23 Statistical Analysis Statistical Package for Social Sciences(SPSS) for Windows 20 (IBM SPSS Inc Chicago USA)program was used for statistical analyses Kolmogorov-Smirnov test was used to determine the distribution of dataContinuous variables with normal distribution were given asmean plusmn standard deviation and continuous variables withoutnormal distribution were given as median interquartile range[IQR] Categorical variables were expressed as numbersand percentage Continuous variables were compared withindependent sample 119905-test ANOVA Mann Whitney 119880 testand Kruskal-Wallis 119867 test where appropriate Chi-squaretest and Fisherrsquos exact chi-square test were used to comparecategorical variables The relationship between the numericparameters was analyzed by Pearson and Spearman cor-relation analysis Stepwise multivariable logistic regressionanalysis was used to identify independent predictors of AArisk A 119901 lt 005 was considered significant for statisticalanalyses

3 Results

A total of 142 individuals were evaluated in the study Nosignificant difference was observed between the AA andcontrol groups with respect to age Hb and BMI (119901 gt005) The native thiol total thiol and the native thioltotalthiol ratio levels were statistically significantly decreased(119901 lt 0001) in the patients with AA compared with thecontrol subjects On the other hand disulphide level andthe ratios of disulphidenative thiol and disulphidetotal thiolwere higher in the AA group than in the control group(119901 lt 0001) The demographics and the thioldisulfidehomeostasis parameters of the patients and healthy controlsare summarized in Table 1

In the appendectomy group patients were further dividedinto two categories nonperforated appendicitis (119899 = 54) andperforated appendicitis (119899 = 17) There was no significantdifference between the two groups in terms of sex BMI andHb (119901 gt 005) The levels of native thiol and total thiol(119901 = 0003 and 119901 = 0005 resp) as well as the nativethioltotal thiol ratio (119901 = 004) were lower in the perforatedgroup when compared to those of the nonperforated groupOn the other hand disulphide level and the ratios of disul-phidenative thiol and disulphidetotal thiol were higher inthe perforated group than in the nonperforated group (119901 =0027 119901 = 0001 and 119901 = 0001 resp) The demographicsand the thioldisulfide homeostasis parameters of the patientsin AA group are summarized in Table 2




(119899 = 71) (119899 = 71)Gender 119899 ()

Male 43 (606) 30 (423) 004Female 28 (394) 41 (577)




(119899 = 17) (119899 = 54)Gender 119899 ()

Male 13 (765) 30 (556) NSFemale 4 (235) 24 (444)





Table3Correlatio


ticsa

ndlabo

ratory

finding

swith

study

popu

latio

n

Varia

bles

Nativethiol

Disu

lphide

Totalthiol

Disu

lphidenativethiol

Disu

lphideto

talthiol

Nativethiolto

talthiol

119903119901

119903119901

119903119901

119903119901

119903119901

119903119901

Groups

Con

trol

Age

(years)

minus0384

0001lowastminus0077

0522minus0378

0001lowast

0125

0297

0131

0277

minus0125

0299

BMI(kgm2

)minus0020

0867minus0106

0378minus0039

0746minus0133

0268

minus0134

0264

0138

0250

WBC

(120583L)

0166

0166

0194

0104

0174

0147

0033

0785

0038

0754

minus0019

0876

CRP(m

gL)

0085

0479

0080

0505

0084

0484minus0016

0893

0003

0979

0035

0774

Hem

oglobin(gdL)

0169

0159minus0057

0639

0164

0171minus0078

0520

minus0076

0529

0079

0512

AA Age

(years)

minus0161

0181

0101

0401minus0268

0024lowast

0192

0109

0179

0135

minus0292

0013lowast

BMI(kgm2

)minus0224

0061

0119

0323minus0154

0201

0128

0286

0136

0260

minus0230

0053

WBC

(120583L)

minus0212

0075minus0017

0888minus0114

0345minus0022

0854

minus0003

0977

0079

0512

CRP(m

gL)

minus0322

000

6lowast0114

0344minus0377

0001lowast

0302

0011lowast

0313

0008lowast

minus0234

0050lowast

Hem

oglobin(gdL)

0094

0434

0139

0247

0120

0320minus0084

0484

minus0089

0461

0041

0733

lowast


nificance

BMIbo

dymassind

ex(kgm2

)WBC

whitebloo

dcellCR

PC-

reactiv

eprotein







4 Discussion









5 Conclusion


Competing Interests




References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















(119899 = 71) (119899 = 71)Gender 119899 ()

Male 43 (606) 30 (423) 004Female 28 (394) 41 (577)




(119899 = 17) (119899 = 54)Gender 119899 ()

Male 13 (765) 30 (556) NSFemale 4 (235) 24 (444)





Table3Correlatio


ticsa

ndlabo

ratory

finding

swith

study

popu

latio

n

Varia

bles

Nativethiol

Disu

lphide

Totalthiol

Disu

lphidenativethiol

Disu

lphideto

talthiol

Nativethiolto

talthiol

119903119901

119903119901

119903119901

119903119901

119903119901

119903119901

Groups

Con

trol

Age

(years)

minus0384

0001lowastminus0077

0522minus0378

0001lowast

0125

0297

0131

0277

minus0125

0299

BMI(kgm2

)minus0020

0867minus0106

0378minus0039

0746minus0133

0268

minus0134

0264

0138

0250

WBC

(120583L)

0166

0166

0194

0104

0174

0147

0033

0785

0038

0754

minus0019

0876

CRP(m

gL)

0085

0479

0080

0505

0084

0484minus0016

0893

0003

0979

0035

0774

Hem

oglobin(gdL)

0169

0159minus0057

0639

0164

0171minus0078

0520

minus0076

0529

0079

0512

AA Age

(years)

minus0161

0181

0101

0401minus0268

0024lowast

0192

0109

0179

0135

minus0292

0013lowast

BMI(kgm2

)minus0224

0061

0119

0323minus0154

0201

0128

0286

0136

0260

minus0230

0053

WBC

(120583L)

minus0212

0075minus0017

0888minus0114

0345minus0022

0854

minus0003

0977

0079

0512

CRP(m

gL)

minus0322

000

6lowast0114

0344minus0377

0001lowast

0302

0011lowast

0313

0008lowast

minus0234

0050lowast

Hem

oglobin(gdL)

0094

0434

0139

0247

0120

0320minus0084

0484

minus0089

0461

0041

0733

lowast


nificance

BMIbo

dymassind

ex(kgm2

)WBC

whitebloo

dcellCR

PC-

reactiv

eprotein







4 Discussion









5 Conclusion


Competing Interests




References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table3Correlatio


ticsa

ndlabo

ratory

finding

swith

study

popu

latio

n

Varia

bles

Nativethiol

Disu

lphide

Totalthiol

Disu

lphidenativethiol

Disu

lphideto

talthiol

Nativethiolto

talthiol

119903119901

119903119901

119903119901

119903119901

119903119901

119903119901

Groups

Con

trol

Age

(years)

minus0384

0001lowastminus0077

0522minus0378

0001lowast

0125

0297

0131

0277

minus0125

0299

BMI(kgm2

)minus0020

0867minus0106

0378minus0039

0746minus0133

0268

minus0134

0264

0138

0250

WBC

(120583L)

0166

0166

0194

0104

0174

0147

0033

0785

0038

0754

minus0019

0876

CRP(m

gL)

0085

0479

0080

0505

0084

0484minus0016

0893

0003

0979

0035

0774

Hem

oglobin(gdL)

0169

0159minus0057

0639

0164

0171minus0078

0520

minus0076

0529

0079

0512

AA Age

(years)

minus0161

0181

0101

0401minus0268

0024lowast

0192

0109

0179

0135

minus0292

0013lowast

BMI(kgm2

)minus0224

0061

0119

0323minus0154

0201

0128

0286

0136

0260

minus0230

0053

WBC

(120583L)

minus0212

0075minus0017

0888minus0114

0345minus0022

0854

minus0003

0977

0079

0512

CRP(m

gL)

minus0322

000

6lowast0114

0344minus0377

0001lowast

0302

0011lowast

0313

0008lowast

minus0234

0050lowast

Hem

oglobin(gdL)

0094

0434

0139

0247

0120

0320minus0084

0484

minus0089

0461

0041

0733

lowast


nificance

BMIbo

dymassind

ex(kgm2

)WBC

whitebloo

dcellCR

PC-

reactiv

eprotein







4 Discussion









5 Conclusion


Competing Interests




References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















4 Discussion









5 Conclusion


Competing Interests




References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















5 Conclusion


Competing Interests




References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Documents

Research Article A Novel Oxidative Stress Mediator in ...downloads.hindawi.com/journals/mi/2016/6761050.pdf · Conclusion . is is the rst study examining the thiol/disulphide homeostasis