Evaluating the levels Salivary acid phosphatase, beta-glucuronidase and cathepsin activities as biochemical markers stress under Psychological Stress and its Relationship with Rumination and Personality Traits.

Shahdokht Rastegar 1*, Mohammad Aberomand2,
1,3 Student Research Committee, 2Toxicology Research Center, Department of Clinical Biochemistry, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

Abstract
Background and purpose: Salivary enzymes are used as a noninvasive biomarker to assess the activity of the sympathetic-adrenal-medullary system. The aim of this study was to evaluate levels of the salivary acid phosphatase, ?-glucuronidase and cathepsin under psychological tention and its connection with rumination and personality traits.
Methods: In a cross-sectional study a total of 60 medical students, who wanted to participate in the ultimate exam, were chosen by simple random sampling. Two months before the exam, in the basal conditions, the Inventory Emotional Control Questionnaire (ECQ) and the Neo-short form were completed. Saliva samples were taken from students in both the basal conditions and under exam stress. Salivary enzymes levels were measured by spectrophotometry and data was analyzed using paired samples t-test, Pearson correlation analysis and Step wise regression.

Results: A significant difference was found between the mean of Salivary enzymes levels in the rest and under exam stress. Also, we found a positive and significant correlation between Salivary enzymes levels with Neuroticism, Agreeableness, Rumination, Extraversion at (P<0.01), (P<0.05) level. Neuroticism, Agreeableness and Rumination predicted 45% of the variance of Salivary acid phosphatase, Neuroticism and Rumination predicted 49 % of the variance of Salivary beta-glucuronidase and Neuroticism, Extraversion and Rumination predicted 38% of the variance of Salivary cathepsin under stress exam.
Conclusion: According to this study, levels of salivary acid phosphatase, beta-glucuronidase and cathepsin may increase in individuals with traits of neuroticism, Extraversion, agreeableness and rumination thought in response to psychological stressors (e.g. exam). Also, measuring salivary enzymes could be used in assessing physiological responses to stress, as a noninvasive method.

Keywords: Salivary enzymes, rumination, personality traits, stress.

INTRODUCTION
Stress as an environmental and psychological stimulus can lead to mental or physiological responses in people. Medical academic exams are considered as acute psychological stress in students , because the performance of the exam in general has future outcomes in the professional life of the students ADDIN EN.CITE <EndNote><Cite><Author>Singh</Author><Year>2012</Year><RecNum>1</RecNum><DisplayText>(1)</DisplayText><record><rec-number>1</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843240″>1</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Singh, Ruchi</author><author>Goyal, Manish</author><author>Tiwari, Sunita</author><author>Ghildiyal, Archana</author><author>Nattu, Shankar M</author><author>Das, Shobha</author></authors></contributors><titles><title>Effect of examination stress on mood, performance and cortisol levels in medical students</title><secondary-title>Indian J Physiol Pharmacol</secondary-title></titles><periodical><full-title>Indian J Physiol Pharmacol</full-title></periodical><pages>48-55</pages><volume>56</volume><number>1</number><dates><year>2012</year></dates><urls></urls></record></Cite></EndNote>(1) Written exams play an basic role in the assessment of learning outcomes and competence of students ADDIN EN.CITE <EndNote><Cite><Author>Singh</Author><Year>2012</Year><RecNum>1</RecNum><DisplayText>(1)</DisplayText><record><rec-number>1</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843240″>1</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Singh, Ruchi</author><author>Goyal, Manish</author><author>Tiwari, Sunita</author><author>Ghildiyal, Archana</author><author>Nattu, Shankar M</author><author>Das, Shobha</author></authors></contributors><titles><title>Effect of examination stress on mood, performance and cortisol levels in medical students</title><secondary-title>Indian J Physiol Pharmacol</secondary-title></titles><periodical><full-title>Indian J Physiol Pharmacol</full-title></periodical><pages>48-55</pages><volume>56</volume><number>1</number><dates><year>2012</year></dates><urls></urls></record></Cite></EndNote>(1) In during exams, medical students experiment a high level of stress and anxiety. The effect of exams stress on the activity of the auto-nervous system and the axis of the hypothalamic-adrenocortical is demonstrated ADDIN EN.CITE <EndNote><Cite><Author>Al-Ayadhi</Author><Year>2005</Year><RecNum>2</RecNum><DisplayText>(2)</DisplayText><record><rec-number>2</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843583″>2</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Al-Ayadhi, Laila Y</author></authors></contributors><titles><title>Neurohormonal changes in medical students during academic stress</title><secondary-title>Ann Saudi Med</secondary-title></titles><periodical><full-title>Ann Saudi Med</full-title></periodical><pages>36-40</pages><volume>25</volume><number>1</number><dates><year>2005</year></dates><urls></urls></record></Cite></EndNote>(2)When stresses Hypothalamic-pituitary-adrenal (HPA) reacts through cortisol secretion and sympathetic-adrenal-medullary (SAM) activated by norepinephrine secretion and increase biomarker stress enzymes ADDIN EN.CITE <EndNote><Cite><Author>Joshi</Author><Year>2012</Year><RecNum>3</RecNum><DisplayText>(3)</DisplayText><record><rec-number>3</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843647″>3</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Joshi, Rachit M</author><author>Sanghavi, Saurin J</author><author>Upadhyaya, Devanshi P</author><author>Chauhan, Ashutosh</author><author>Halvadia, Shital</author></authors></contributors><titles><title>Effect of examination stress on the plasma cortisol level</title><secondary-title>National Journal of Medical Research</secondary-title></titles><periodical><full-title>National Journal of Medical Research</full-title></periodical><pages>435-438</pages><volume>2</volume><number>4</number><dates><year>2012</year></dates><urls></urls></record></Cite></EndNote>(3)Newly,utilize biological fluid like oral fluid or saliva as a diagnostic instrument or specific marker for systemic conditions has increased exponentially ADDIN EN.CITE <EndNote><Cite><Author>Kinney</Author><Year>2007</Year><RecNum>4</RecNum><DisplayText>(4)</DisplayText><record><rec-number>4</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843700″>4</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Kinney, Janet S</author><author>Ramseier, Christoph A</author><author>Giannobile, William V</author></authors></contributors><titles><title>Oral fluid–based biomarkers of alveolar bone loss in periodontitis</title><secondary-title>Annals of the New York Academy of Sciences</secondary-title></titles><periodical><full-title>Annals of the New York Academy of Sciences</full-title></periodical><pages>230-251</pages><volume>1098</volume><number>1</number><dates><year>2007</year></dates><isbn>1749-6632</isbn><urls></urls></record></Cite></EndNote>(4)Salivary components such as enzymes, immunoglobulin’s, inorganic materials and ions have different effects on mouth homeostasis and any change in their balance may lead to oral periodontal diseases including dental caries ADDIN EN.CITE ;EndNote;;Cite;;Author;Kaufman;/Author;;Year;2000;/Year;;RecNum;5;/RecNum;;DisplayText;(5, 6);/DisplayText;;record;;rec-number;5;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843753″;5;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Kaufman, Eliaz;/author;;author;Lamster, Ira B;/author;;/authors;;/contributors;;titles;;title;Analysis of saliva for periodontal diagnosis: a review;/title;;secondary-title;Journal of clinical periodontology;/secondary-title;;/titles;;periodical;;full-title;Journal of clinical periodontology;/full-title;;/periodical;;pages;453-465;/pages;;volume;27;/volume;;number;7;/number;;dates;;year;2000;/year;;/dates;;isbn;0303-6979;/isbn;;urls;;/urls;;/record;;/Cite;;Cite;;Author;Lamster;/Author;;Year;2003;/Year;;RecNum;6;/RecNum;;record;;rec-number;6;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843796″;6;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Lamster, Ira B;/author;;author;Kaufman, Eliaz;/author;;author;Grbic, John T;/author;;author;Winston, Leslie J;/author;;author;Singer, Robert E;/author;;/authors;;/contributors;;titles;;title;??glucuronidase activity in saliva: relationship to clinical periodontal parameters;/title;;secondary-title;Journal of periodontology;/secondary-title;;/titles;;periodical;;full-title;Journal of periodontology;/full-title;;/periodical;;pages;353-359;/pages;;volume;74;/volume;;number;3;/number;;dates;;year;2003;/year;;/dates;;isbn;0022-3492;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(5, 6)Research evidence shows increase levels of lysosomal enzymes such as acid phosphatase, beta-glucuronidase and cathepsin not only the responsiveness to physical agents such as shock, breakage of sound walls, waves, heat and cold, as well as psychological stress such as anxiety, stress, fatigue, and exam ADDIN EN.CITE ;EndNote;;Cite;;Author;Koh;/Author;;Year;2014;/Year;;RecNum;7;/RecNum;;DisplayText;(7);/DisplayText;;record;;rec-number;7;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843841″;7;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Koh, David;/author;;author;Ng, Vivian;/author;;author;Naing, Lin;/author;;/authors;;/contributors;;titles;;title;Alpha amylase as a salivary biomarker of acute stress of venepuncture from periodic medical examinations;/title;;secondary-title;Frontiers in public health;/secondary-title;;/titles;;periodical;;full-title;Frontiers in public health;/full-title;;/periodical;;pages;121;/pages;;volume;2;/volume;;dates;;year;2014;/year;;/dates;;isbn;2296-2565;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(7)Previous studies showed that exam stress in addition to, increase blood pressure and anxiety can increase salivary bio markers such as cortisol and immunoglobulin ADDIN EN.CITE ;EndNote;;Cite;;Author;Takatsuji;/Author;;Year;2008;/Year;;RecNum;8;/RecNum;;DisplayText;(8);/DisplayText;;record;;rec-number;8;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843904″;8;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Takatsuji, Koichi;/author;;author;Sugimoto, Yoshie;/author;;author;Ishizaki, Shoko;/author;;author;Ozaki, Yasuka;/author;;author;Matsuyama, Etsuko;/author;;author;Yamaguchi, Yukari;/author;;/authors;;/contributors;;titles;;title;The effects of examination stress on salivary cortisol, immunoglobulin A, and chromogranin A in nursing students;/title;;secondary-title;Biomedical Research;/secondary-title;;/titles;;periodical;;full-title;Biomedical Research;/full-title;;/periodical;;pages;221-224;/pages;;volume;29;/volume;;number;4;/number;;dates;;year;2008;/year;;/dates;;isbn;0388-6107;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(8)These studies show that exam stress increases the anxiety and salivary cortisol levels of male students significantly, but decreased their salivary testosterone level ADDIN EN.CITE ;EndNote;;Cite;;Author;Choi;/Author;;Year;2012;/Year;;RecNum;9;/RecNum;;DisplayText;(9);/DisplayText;;record;;rec-number;9;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533843951″;9;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Choi, JC;/author;;author;Chung, MI;/author;;author;Lee, YD;/author;;/authors;;/contributors;;titles;;title;Modulation of pain sensation by stress?related testosterone and cortisol;/title;;secondary-title;Anaesthesia;/secondary-title;;/titles;;periodical;;full-title;Anaesthesia;/full-title;;/periodical;;pages;1146-1151;/pages;;volume;67;/volume;;number;10;/number;;dates;;year;2012;/year;;/dates;;isbn;0003-2409;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(9)However, there are little information on the relationship between acid phosphatase, beta-glucuronidase and cathepsin enzymes with the exam psychological stress. Only one study showed increased activity of acid phosphatase and beta-glucuronidase enzymes under stress test conditions ADDIN EN.CITE ;EndNote;;Cite;;Author;Layik;/Author;;Year;2000;/Year;;RecNum;10;/RecNum;;DisplayText;(10);/DisplayText;;record;;rec-number;10;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844017″;10;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Layik, Meral;/author;;author;Yamalik, Nermin;/author;;author;Çaglayan, Feriha;/author;;author;Kilinç, Kamer;/author;;author;Etikan, Ilker;/author;;author;Eratalay, Kenan;/author;;/authors;;/contributors;;titles;;title;Analysis of human gingival tissue and gingival crevicular fluid ?-glucuronidase activity in specific periodontal diseases;/title;;secondary-title;Journal of periodontology;/secondary-title;;/titles;;periodical;;full-title;Journal of periodontology;/full-title;;/periodical;;pages;618-624;/pages;;volume;71;/volume;;number;4;/number;;dates;;year;2000;/year;;/dates;;isbn;0022-3492;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(10)conforming to the previous proof, it is not obvious whether the activity of these enzymes under academic examine stress, principally written exams will be increased or not? For this reason, we decided to survey the activity of these enzymes under written exam stress. On the other hand, the reaction to the stress of the exam can be impress by personality traits and psychological factors. These study, probe the communication between rumination and personality traits with level activity enzymes. The rumination as a structure related to psychological stress can trigger and sustain physiological activities associated with stress ADDIN EN.CITE ;EndNote;;Cite;;Author;Ohura;/Author;;Year;2012;/Year;;RecNum;11;/RecNum;;DisplayText;(11, 12);/DisplayText;;record;;rec-number;11;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844079″;11;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Ohura, Kiyoshi;/author;;author;Nozaki, Tadashige;/author;;author;Shinohara, Mitsuko;/author;;author;Daito, Kiyoshi;/author;;author;Sonomoto, Mie;/author;;author;Daito, Michiharu;/author;;/authors;;/contributors;;titles;;title;Utility of salivary biomarker for stress induced by dental treatment;/title;;secondary-title;Japanese Dental Science Review;/secondary-title;;/titles;;periodical;;full-title;Japanese Dental Science Review;/full-title;;/periodical;;pages;14-17;/pages;;volume;48;/volume;;number;1;/number;;dates;;year;2012;/year;;/dates;;isbn;1882-7616;/isbn;;urls;;/urls;;/record;;/Cite;;Cite;;Author;Zoccola;/Author;;Year;2012;/Year;;RecNum;12;/RecNum;;record;;rec-number;12;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844123″;12;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Zoccola, Peggy M;/author;;author;Dickerson, Sally S;/author;;/authors;;/contributors;;titles;;title;Assessing the relationship between rumination and cortisol: A review;/title;;secondary-title;Journal of Psychosomatic Research;/secondary-title;;/titles;;periodical;;full-title;Journal of Psychosomatic Research;/full-title;;/periodical;;pages;1-9;/pages;;volume;73;/volume;;number;1;/number;;dates;;year;2012;/year;;/dates;;isbn;0022-3999;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(11, 12)The rumination is a process constant and involuntary thinking about negative events, which include special phase of depression, such as generate negative thinking, debility of executive control and persistent on the mental negative situation. The rumination is important in survival and severity of stress and its enhancement are connected with a rise in stress ADDIN EN.CITE ;EndNote;;Cite;;Author;Smith;/Author;;Year;2009;/Year;;RecNum;13;/RecNum;;DisplayText;(13, 14);/DisplayText;;record;;rec-number;13;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844182″;13;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Smith, Jeannette M;/author;;author;Alloy, Lauren B;/author;;/authors;;/contributors;;titles;;title;A roadmap to rumination: A review of the definition, assessment, and conceptualization of this multifaceted construct;/title;;secondary-title;Clinical psychology review;/secondary-title;;/titles;;periodical;;full-title;Clinical psychology review;/full-title;;/periodical;;pages;116-128;/pages;;volume;29;/volume;;number;2;/number;;dates;;year;2009;/year;;/dates;;isbn;0272-7358;/isbn;;urls;;/urls;;/record;;/Cite;;Cite;;Author;Van Vugt;/Author;;Year;2012;/Year;;RecNum;14;/RecNum;;record;;rec-number;14;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844228″;14;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Van Vugt, Marieke Karlijn;/author;;author;Hitchcock, Peter;/author;;author;Shahar, Ben;/author;;author;Britton, Willoughby;/author;;/authors;;/contributors;;titles;;title;The effects of mindfulness-based cognitive therapy on affective memory recall dynamics in depression: a mechanistic model of rumination;/title;;secondary-title;Frontiers in human neuroscience;/secondary-title;;/titles;;periodical;;full-title;Frontiers in human neuroscience;/full-title;;/periodical;;pages;257;/pages;;volume;6;/volume;;dates;;year;2012;/year;;/dates;;isbn;1662-5161;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(13, 14)The recently studies, confirm the communication between rumination and responses of cortisol ADDIN EN.CITE ;EndNote;;Cite;;Author;Zoccola;/Author;;Year;2012;/Year;;RecNum;15;/RecNum;;DisplayText;(12);/DisplayText;;record;;rec-number;15;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844295″;15;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Zoccola, Peggy M;/author;;author;Dickerson, Sally S;/author;;/authors;;/contributors;;titles;;title;Assessing the relationship between rumination and cortisol: A review;/title;;secondary-title;Journal of Psychosomatic Research;/secondary-title;;/titles;;periodical;;full-title;Journal of Psychosomatic Research;/full-title;;/periodical;;pages;1-9;/pages;;volume;73;/volume;;number;1;/number;;dates;;year;2012;/year;;/dates;;isbn;0022-3999;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(12)and salivary alpha-amylase ADDIN EN.CITE ;EndNote;;Cite;;Author;Afrisham;/Author;;Year;2015;/Year;;RecNum;16;/RecNum;;DisplayText;(15);/DisplayText;;record;;rec-number;16;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844345″;16;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Afrisham, Reza;/author;;author;Sadegh-Nejadi, Sahar;/author;;author;SoliemaniFar, Omid;/author;;author;Abromand, Mohammad;/author;;author;Kooti, Wesam;/author;;author;Najjar Asl, Sedigheh;/author;;author;Khaneh Keshi, Ali;/author;;/authors;;/contributors;;titles;;title;Evaluating the salivary alpha-amylase level under psychological stress and its relationship with rumination and the five personality traits;/title;;secondary-title;Journal of Mazandaran University of Medical Sciences;/secondary-title;;/titles;;periodical;;full-title;Journal of Mazandaran University of Medical Sciences;/full-title;;/periodical;;pages;22-33;/pages;;volume;25;/volume;;number;126;/number;;dates;;year;2015;/year;;/dates;;urls;;/urls;;/record;;/Cite;;/EndNote;(15)to stress, but empirical research lacking on the relationship between salivary enzymes and the trait of rumination, as a psychological characteristic associated with stress. It appears that personality can be better explained by the Five Factor Model of personality, which offers a normal structure of personality traits using the Personality Inventory NEO-PI. This model has been widely accepted as the most prominent structure of personality. According to this model, the interpersonal variance in personality traits is explained through five major dimensions of personality, such as extraversion, agreeableness, conscientiousness, neuroticism, and openness to experience ADDIN EN.CITE ;EndNote;;Cite;;Author;Costa;/Author;;Year;2008;/Year;;RecNum;17;/RecNum;;DisplayText;(16);/DisplayText;;record;;rec-number;17;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844393″;17;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Costa, Paul T;/author;;author;McCrae, Robert R;/author;;/authors;;/contributors;;titles;;title;The revised neo personality inventory (neo-pi-r);/title;;secondary-title;The SAGE handbook of personality theory and assessment;/secondary-title;;/titles;;periodical;;full-title;The SAGE handbook of personality theory and assessment;/full-title;;/periodical;;pages;179-198;/pages;;volume;2;/volume;;number;2;/number;;dates;;year;2008;/year;;/dates;;urls;;/urls;;/record;;/Cite;;/EndNote;(16)In previous studies using this model reported a negative relationship between neuroticism with cortisol ADDIN EN.CITE ;EndNote;;Cite;;Author;Nater;/Author;;Year;2010;/Year;;RecNum;18;/RecNum;;DisplayText;(17);/DisplayText;;record;;rec-number;18;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844440″;18;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Nater, Urs M;/author;;author;Hoppmann, Christiane;/author;;author;Klumb, Petra L;/author;;/authors;;/contributors;;titles;;title;Neuroticism and conscientiousness are associated with cortisol diurnal profiles in adults—Role of positive and negative affect;/title;;secondary-title;Psychoneuroendocrinology;/secondary-title;;/titles;;periodical;;full-title;Psychoneuroendocrinology;/full-title;;/periodical;;pages;1573-1577;/pages;;volume;35;/volume;;number;10;/number;;dates;;year;2010;/year;;/dates;;isbn;0306-4530;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(17) and salivary alpha-amylase as well as a positive correlation between agreeableness with salivary alpha-amylase ADDIN EN.CITE ;EndNote;;Cite;;Author;Inukai;/Author;;Year;2010;/Year;;RecNum;19;/RecNum;;DisplayText;(18);/DisplayText;;record;;rec-number;19;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844497″;19;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Inukai, Keigo;/author;;author;Shinada, Mizuho;/author;;author;Tanida, Shigehito;/author;;author;Takahashi, Chisato;/author;;author;Mifune, Nobuhiro;/author;;author;Takagishi, Haruto;/author;;author;Horita, Yutaka;/author;;author;Hashimoto, Hirofumi;/author;;author;Yokota, Kunihiro;/author;;author;Kameda, Tatsuya;/author;;/authors;;/contributors;;titles;;title;Salivary alpha-amylase levels and big five personality factors in adults;/title;;secondary-title;Neuroendocrinology Letters;/secondary-title;;/titles;;periodical;;full-title;Neuroendocrinology Letters;/full-title;;/periodical;;pages;771;/pages;;volume;31;/volume;;number;6;/number;;dates;;year;2010;/year;;/dates;;isbn;0172-780X;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(18)For this reason, we have tried to study the relationship between rumination and salivary acid phosphatase, cathepsin and beta-glucuronidase under psychological stress. What is discussed, is whether the personal differences in biological response to stress are, reflecting differences in basic human personality traits or not? To date,in advanced studies using different measurement tools, reported the higher levels of personality dimension that include connection between personality variables and stress such as hardiness, sensation seeking, achievement motivation, extraversion and neuroticism. ADDIN EN.CITE ;EndNote;;Cite;;Author;Hu;/Author;;Year;2014;/Year;;RecNum;20;/RecNum;;DisplayText;(19);/DisplayText;;record;;rec-number;20;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844551″;20;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Hu, Emily;/author;;author;Koucky, Ellen M;/author;;author;Brown, Wilson J;/author;;author;Bruce, Steven E;/author;;author;Sheline, Yvette I;/author;;/authors;;/contributors;;titles;;title;The role of rumination in elevating perceived stress in posttraumatic stress disorder;/title;;secondary-title;Journal of interpersonal violence;/secondary-title;;/titles;;periodical;;full-title;Journal of interpersonal violence;/full-title;;/periodical;;pages;1953-1962;/pages;;volume;29;/volume;;number;10;/number;;dates;;year;2014;/year;;/dates;;isbn;0886-2605;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(19)In fact, changes in the level of lysosomal enzymes can affect the rumination characteristics of individuals and details rumination may be associated with increased levels of salivary enzymes. So it is assumed that changes in acid phosphatase, beta-glucuronidase and cathepsin enzymes can affected rumination characteristics of people. In other words, the characteristics of rumination may be associated with increased levels of the enzyme acid phosphatase, beta-glucuronidase and cathepsin under stress. Therefore, we investigated the relationship between rumination and personality traits with salivary acid phosphatase, beta-glucuronidase and cathepsin under psychological stress in medical students.

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Participants: This cross-sectional and descriptive study and the population included all the government medical students at Ahvaz University of Medical Sciences who wanted to participate in a final exam. In this study, 60 students (30 women and 30 men) were selected, using the Cochran formula and the simple random sampling method. The inclusion criteria for the study included the absence of infectious disease; autoimmune; hereditary immune deficiency; mental illness; lack of drugs to suppress the immune system (immunosuppressive); no vaccine or blood; endocrine diseases such as diabetes; non-smoking; and lack of surgery during two months before saliva sampling. Due to the students being freshmen, thus, the exam was considered a psychologically stressful situation. Data were collected using the personality Neo-short form (FFI-NEO) and Emotional Control Questionnaire (ECQ) ADDIN EN.CITE ;EndNote;;Cite;;Author;Afrisham;/Author;;Year;2015;/Year;;RecNum;21;/RecNum;;DisplayText;(15, 20);/DisplayText;;record;;rec-number;21;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844605″;21;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Afrisham, Reza;/author;;author;Sadegh-Nejadi, Sahar;/author;;author;SoliemaniFar, Omid;/author;;author;Abromand, Mohammad;/author;;author;Kooti, Wesam;/author;;author;Najjar Asl, Sedigheh;/author;;author;Khaneh Keshi, Ali;/author;;/authors;;/contributors;;titles;;title;Evaluating the salivary alpha-amylase level under psychological stress and its relationship with rumination and the five personality traits;/title;;secondary-title;Journal of Mazandaran University of Medical Sciences;/secondary-title;;/titles;;periodical;;full-title;Journal of Mazandaran University of Medical Sciences;/full-title;;/periodical;;pages;22-33;/pages;;volume;25;/volume;;number;126;/number;;dates;;year;2015;/year;;/dates;;urls;;/urls;;/record;;/Cite;;Cite;;Author;Ghafourian-Boroujerdnia;/Author;;Year;2014;/Year;;RecNum;22;/RecNum;;record;;rec-number;22;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844644″;22;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Ghafourian-Boroujerdnia, Mehri;/author;;author;Shiravi, Zahra;/author;;author;Hamid, Najmeh;/author;;author;Hemmati, Ali-Asghar;/author;;author;Kooti, Wesam;/author;;/authors;;/contributors;;titles;;title;The Relationship of Hardiness and Immune System Cells;/title;;secondary-title;Journal of Isfahan Medical School;/secondary-title;;/titles;;periodical;;full-title;Journal of Isfahan Medical School;/full-title;;/periodical;;volume;31;/volume;;number;260;/number;;dates;;year;2014;/year;;/dates;;isbn;1027-7595;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(15, 20)
Questionnaires
The personality Neo-short form: In this study, McCrae and Costa’s 60-items questionnaire (1992) was used. This scale measures the five major personality traits, which include: Neuroticism (N), Extraversion (E), Agreeableness (A), Openness to Experience (O), and conscientiousness (C). Each of these traits can be measured with 12 questions. The answer to the questionnaire is set based on a five-point Likert scale: completely disagree (0); disagree (1); indifferent (2); agree (3); and completely agree (4). Based on Cronbach’s alpha coefficients by Costa and McCrae, the internal consistency of this scale for traits of N, E, O, A, and C was 0.86, 0.77, 0.73, 0.68, and 0.81, respectively ADDIN EN.CITE ;EndNote;;Cite;;Author;Digman;/Author;;Year;1997;/Year;;RecNum;23;/RecNum;;DisplayText;(21);/DisplayText;;record;;rec-number;23;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844729″;23;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Digman, John M;/author;;/authors;;/contributors;;titles;;title;Higher-order factors of the Big Five;/title;;secondary-title;Journal of personality and social psychology;/secondary-title;;/titles;;periodical;;full-title;Journal of personality and social psychology;/full-title;;/periodical;;pages;1246;/pages;;volume;73;/volume;;number;6;/number;;dates;;year;1997;/year;;/dates;;isbn;1939-1315;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(21)In this study, using Cronbach’s alpha, the coefficient of reliability of the questionnaire calculated for the traits of N, E, O, A, and C was 0.76, 0.78, 0.77, 0.72, and 0.81, respectively.

Emotional Control Questionnaire: The questionnaire was developed by Roger and Nesshoever (1987) and was renewed by Roger and Najarian (1989). The ECQ measures four scales: Emotional Inhibition, aggression control, ruminants or rehearsing and benign control measures; it has 56 items. Each subscale has 14 questions. The score of a person in each subscale ranges from zero to 14 and in the total scale ranges from zero to 56. High scores indicate greater emotional control ADDIN EN.CITE <EndNote><Cite><Author>Roger</Author><Year>1998</Year><RecNum>24</RecNum><DisplayText>(22)</DisplayText><record><rec-number>24</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844781″>24</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Roger, Derek</author><author>Najarian, Bahman</author></authors></contributors><titles><title>The relationship between emotional rumination and cortisol secretion under stress</title><secondary-title>Personality and Individual Differences</secondary-title></titles><periodical><full-title>Personality and Individual Differences</full-title></periodical><pages>531-538</pages><volume>24</volume><number>4</number><dates><year>1998</year></dates><urls></urls></record></Cite></EndNote>(22)The validity of the questionnaire, using internal consistency, and Cronbach’s alpha coefficient calculated by Rafineya et al ADDIN EN.CITE ;EndNote;;Cite;;Author;Rafineya;/Author;;Year;2006;/Year;;RecNum;25;/RecNum;;DisplayText;(23);/DisplayText;;record;;rec-number;25;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844825″;25;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Rafineya, P;/author;;author;Rasolzadeh Tabatabaei, S;/author;;author;Azad Falah, P;/author;;/authors;;/contributors;;titles;;title;The relation between emotional expression and mental health in students;/title;;secondary-title;J Psychol;/secondary-title;;/titles;;periodical;;full-title;J Psychol;/full-title;;/periodical;;pages;85-104;/pages;;volume;10;/volume;;number;10;/number;;dates;;year;2006;/year;;/dates;;urls;;/urls;;/record;;/Cite;;/EndNote;(23)for each subscale of emotional inhibition, aggression control, rumination, and benign control were 0.70, 0.76, 0.77, and 0.58 respectively and its total validity was 0.68. In the study, the reliability coefficient obtained using Cronbach’s alpha coefficient for the total scale was 0.75 and for each of the subscales of emotional inhibition, aggression control, rumination, and benign control was 0.68, 0.66, 0.65 and 0.74, respectively.
Methods: To conduct the research in the first phase, two months before the exam (basal condition), and all participants completed the Personality NEO- short form questionnaire and emotional control Questionnaires (ECQ). Then, two days later, saliva samples were taken to measure salivary enzymes in the basal conditions. To collect the saliva sample, the oral cavity was first washed with water and after 10 minutes, patients were asked to swallow their saliva. Then, for 2 minutes, put the saliva in your mouth and spitting it into a sterile container (the amount of saliva collected from each person was 3 ml). Saliva collected in sterile tubes containing dry ice was immediately transported to the laboratory. In order to remove mucus, saliva by centrifugation at 11,000 revolutions refrigerated for 10 minutes was isolated. The specimens were then stored in a -70 freeze ADDIN EN.CITE <EndNote><Cite><Author>Azizi</Author><Year>2011</Year><RecNum>26</RecNum><DisplayText>(24)</DisplayText><record><rec-number>26</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844872″>26</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Azizi, Arash</author><author>Ranjbari, Ardeshir</author><author>Ghafari, Mohammad Ali</author><author>Jahan, Fatemeh</author></authors></contributors><titles><title>Comparative evaluation of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) levels in periodontal diseases</title><secondary-title>Journal of Isfahan Dental School</secondary-title></titles><periodical><full-title>Journal of Isfahan Dental School</full-title></periodical><volume>7</volume><number>3</number><dates><year>2011</year></dates><isbn>1735-255X</isbn><urls></urls></record></Cite></EndNote>(24)In the second phase, once again to measure salivary enzymes during exam stress, saliva samples were taken at the start of the exam. In both phases of sampling, all participants were asked not to take caffeine the next night before sampling and 1 hour before the test to avoid eating any food in order to avoid interference in the sampling and measurement of salivary enzymes ADDIN EN.CITE <EndNote><Cite><Author>Afrisham</Author><Year>2015</Year><RecNum>27</RecNum><DisplayText>(15, 22)</DisplayText><record><rec-number>27</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533844955″>27</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Afrisham, Reza</author><author>Sadegh-Nejadi, Sahar</author><author>SoliemaniFar, Omid</author><author>Abromand, Mohammad</author><author>Kooti, Wesam</author><author>Najjar Asl, Sedigheh</author><author>Khaneh Keshi, Ali</author></authors></contributors><titles><title>Evaluating the salivary alpha-amylase level under psychological stress and its relationship with rumination and the five personality traits</title><secondary-title>Journal of Mazandaran University of Medical Sciences</secondary-title></titles><periodical><full-title>Journal of Mazandaran University of Medical Sciences</full-title></periodical><pages>22-33</pages><volume>25</volume><number>126</number><dates><year>2015</year></dates><urls></urls></record></Cite><Cite><Author>Roger</Author><Year>1998</Year><RecNum>28</RecNum><record><rec-number>28</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845006″>28</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Roger, Derek</author><author>Najarian, Bahman</author></authors></contributors><titles><title>The relationship between emotional rumination and cortisol secretion under stress</title><secondary-title>Personality and Individual Differences</secondary-title></titles><periodical><full-title>Personality and Individual Differences</full-title></periodical><pages>531-538</pages><volume>24</volume><number>4</number><dates><year>1998</year></dates><urls></urls></record></Cite></EndNote>(15, 22)Sampling was performed in the same circumstances and particular time (10 am). The level of activity of the acid phosphatase, beta-glucuronidase and cathepsin enzymes was measured by manual and enzymatic (END POINT) methods. The level of enzymes activity (U/dL) was measured by the Kinetic method and with )phenolphthalein beta-D-glucuronide(, (Paranitrofenil phosphate (PNPP) and (N-SUCCINYL-ALA-ALA-PRO-PHE-P-NITROANILIDE) as substrate beta-glucuronidase, acid phosphatase and cathepsin enzymes .The light absorption which matches the activity of ensymes was assessed by a spectrophotometer at the 405 nm (acid phosphatase) and 540 nm (beta-glucuronidase and cathepsin) wavelength ADDIN EN.CITE <EndNote><Cite><Author>Louati</Author><Year>2010</Year><RecNum>29</RecNum><DisplayText>(25)</DisplayText><record><rec-number>29</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845053″>29</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Louati, Hanen</author><author>Zouari, Nacim</author><author>Fendri, Ahmed</author><author>Gargouri, Youssef</author></authors></contributors><titles><title>Digestive amylase of a primitive animal, the scorpion: Purification and biochemical characterization</title><secondary-title>Journal of Chromatography B</secondary-title></titles><periodical><full-title>Journal of Chromatography B</full-title></periodical><pages>853-860</pages><volume>878</volume><number>11-12</number><dates><year>2010</year></dates><isbn>1570-0232</isbn><urls></urls></record></Cite></EndNote>(25)
Ethical principles: This project was approved by the code of ethics of ajums.REC in student research committee of Ahvaz Jundishapur University of Medical Sciences.

Statistical analyses
Statistical analysis was performed with SPSS version 18. Data was analyzed using descriptive statistical methods including mean and standard deviation, and multivariate analysis of variance with repeated measures for the effects of within-subjects factor (salivary enzymes levels measured at 2 steps) and interaction with gender, paired samples t-test to compare the difference in salivary enzymes of the basal conditions and under exam stress, and the Pearson correlation coefficient and stepwise regression analysis. The significance level was less than ( 0.05).

RESULTS
Demographic and Clinical Characteristics: The present study was conducted on 60 students (30 men and 30 women). The study participants ranged in age from 19–22 years, and mean age 19.7±0.917 years. The summary statistics concerning the demographic characteristics and questionnaires data for all study participants (n=60) are presented in Table 1. Mean and standard deviation for the neuroticism, extraversion, openness to experience, agreeableness, conscientiousness, and rumination were 36.4±6.54, 31.55±4.68, 37.46±3.59, 41.38±4.08, 43.81±7.05and 8.73±2.39 respectively (Table 1).
Comparison of salivary enzymes levels in the basal conditions and under exam stress: According to the results of the t-test of the coupled specimens in Table 2, there is a significant difference between the mean amount of acid phosphatase enzymes (T = -2.163 ? P=0/01 ), beta-glucuronidase(T = -3.231 ? P=0/016 ) and salivary cathepsin (T = -4.445 ? P=0/035 ) in terms of rest and test conditions. The significance level of P<0.05 indicates that salivary enzymes of students under stress test conditions are significantly higher than rest or control conditions. Table 2 shows the average activity of acid phosphatase, beta-glucuronidase and cathepsin enzymes in saliva of students at rest and in the exam.
Correlation of five personality traits and rumination with salivary enzymes: The results of Matrix Correlation Table 3 show that, there is a positive and significant correlation between rumination (p<0.01, r=0.48), neuroticism (p<0.05, r=0.34 ) and agreeableness (p<0.01, r=0.284) scales with salivary acid phosphatase, also between rumination (p<0.01, r=0.41) and neuroticism (p<0.01, r=0.261) scales with salivary ?eta-glucuronidase and between rumination (p<0.05, r=0.35 ), extraversion(0.01, r=0.30) and neuroticism(p<0.05, r=0.31) scales with salivary cathepsin at 0.05 and 0.01 levels respectively. Before using the regression analysis, the existence of single-variable pertinent and overlapping data was first investigated using the plaque box. The results showed that there are 12 overlapping data that was replaced by the nearest available data. The uniform distribution of variables was studied using a rectangular plot. The results showed that the data distribution is also uniform. In addition, the condensation distribution graph, the observed and expected values, showed a 45-degree gradient, and all points were on the line, indicating the normal distribution of the remainders. The same assumption of variances was investigated by charting the standardized regression residues against the predicted regression standard values. The points were randomly scattered, indicating the same variance. After analyzing the assumptions of multiple regression and ensuring that they were established to determine the share predictor variables in the Variable variance explanation, a multiple stepwise regression was used. In order to predict Salivary enzymes on the exam condition, a Stepwise regression analysis was used in Table 4. The results of the regression analysis in Table 4 indicated that trait Rumination, Agreeableness and Neuroticism with coefficients (?); 0.34, 0.37, 0.48 respectively, were able to predict 45% of acid phosphatase, also Rumination and Neuroticism with coefficients (?); 0.53 and 0.38 and Rumination, extravertion and Neuroticism with coefficients (?); 0.33, 0.31, 0.48 respectively, were able to predict 49% of beta-glucuronidase and 38% of cathepsin enzymes under the stress test conditions.
Table 1. Demographic and clinical characteristics of the study subjects (N=60).

Variable Mean ± SD (N,%)
Age 19.7± 0.917
Sex
Male 30 (50 %)
Female 30 (50%) Rumination 8.73 ± 2.39
Emotional inhibition 6.11 ± 2.40
Aggression control
Benign control 8.10 ± 2.25
8.26 ± 2.57
Neuroticism 36.46 ± 6.54
Extraversion 31.55 ±4.68
Openness 37.46 ± 3.59
Agreeableness 41.38 ± 4.08
Consciousness 43.81 ± 7.05
Table 2: Mean and standard deviation of salivary enzymes in the two months before and under test conditions in the whole sample.
Enzymes College BSE
9575804254500-29845425450
Mean ± SD SEE
Mean ± SD df t P -value
Acid phosphatase
Beta-glucuronidase
Cathepsin (N=60)
(N=60)
(N=60) 4.98 ± 1.10
6.55 ± 1.348
21.06 ± 4.47 10.05 ± 1.69
15.807 ± 1.49
54.81 ± 3.49 59
59
59 -2.163
-3.231
-4.445 0.01*
0.016*
0.035*
*p<0.05. BSE: Basal Salivary Enzymes, SEE: Salivary Enzymes under Exam.

Table 3. Correlation of FPT, rumination with salivary enzymes level (u/mL) in basal and exam situations (N=60)
Variable Acid phosphatase
61595825500
BSE SEE
Beta glucuronidase
59690825500
BSE SEE
Cathepin
1974858001000
BSE SEE
FPT Neuroticism
Extraversion
Openness
Agreeableness
Consciousness -0.033
-0.084
-0.005
0.027
0.096 0.34*
0.076
0.018
0.281**
0.236 -0.143
-0.012
0.069*
-0.019
0.171 0.261**
– 0.125
-0.037
0.088
0.045 0.160
0.179
-0.147
-0.094
0.074 0.310*
0.30**
0. 160
0.014
0.053
ECQ Rumination
Emotional inhibition -0.167
-0.303* 0.48**
-0.108
0.121
0.002 -0.23
0.066
-0.035
.219 0.41**
– 0.103
0.001
0.126 -0.028
0.87
-0.153
-0.152 0.35*
-0.151
0.096
0.067
Aggression control 0.21 Benign control -0.091 *p<0.05, **p<0.001. BSE: Basal Salivary Enzymes, SEE: Salivary Enzymes under Exam, FPT: five personality traits, ECQ: Emotional Control Questionnair.
Table 5. Stepwise regression analysis to predict salivary enzymes (Acid phosphatase, Beta-glucuronidase and Cathepsin) (u/mL) under exam condition (N=60)
Enzymes Mode Variable MR R2 F b p
Acid phosphatase 1
2
3 Rumination
Agreeableness
Neuroticism 0.49
0.33
0.56 0.23
0.34
0.45 9.87
8.04
12.52 0.34
0.37
0.48 0.006*
0.003*
0.005*
Beta-glucuronidase 1
2 Rumination
Neuroticism 0.70
0.63 0.38
0.49 8.57
9.65 0.53
0.38 0.025*
0.018*
Cathepsin 1
2
3 Rumination
Extraversion
Neuroticism 0.48
0.35
0.45 0.24
0.35
0.38 9.15
8.52
11.21 0.33
0.31
0.48 0.009*
0.008*
0.012*
DISCUSSION
The purpose of this study was to investigate the activity of salivary acid phosphatase, beta-glucuronidase and cathepsin under psychological stress and its relationship with rumination and five personality traits in medical students. In the present study, the amount of activity of acid phosphatase, beta-glucuronidase and cathepsin in saliva samples of students was measured in two resting and exam times, which was significantly higher than the rest period at the exam time. It seems cause of this phenomenon is the destruction of lysosomal cellular organelles and the release of intracellular material, especially enzymes into tissues and biological fluids including saliva and serum ADDIN EN.CITE <EndNote><Cite><Author>Aits</Author><Year>2013</Year><RecNum>30</RecNum><DisplayText>(26)</DisplayText><record><rec-number>30</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845114″>30</key></foreign-keys><ref-type name=”Generic”>13</ref-type><contributors><authors><author>Aits, Sonja</author><author>Jäättelä, Marja</author></authors></contributors><titles><title>Lysosomal cell death at a glance</title></titles><dates><year>2013</year></dates><publisher>The Company of Biologists Ltd</publisher><isbn>0021-9533</isbn><urls></urls></record></Cite></EndNote>(26)In fact, increase in the level of stress and anxiety in the first year of medical students has been reported ADDIN EN.CITE <EndNote><Cite><Author>Susman</Author><Year>2010</Year><RecNum>31</RecNum><DisplayText>(27)</DisplayText><record><rec-number>31</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845158″>31</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Susman, Elizabeth J</author><author>Dockray, Samantha</author><author>Granger, Douglas A</author><author>Blades, Keeva T</author><author>Randazzo, William</author><author>Heaton, Jodi A</author><author>Dorn, Lorah D</author></authors></contributors><titles><title>Cortisol and alpha amylase reactivity and timing of puberty: Vulnerabilities for antisocial behaviour in young adolescents</title><secondary-title>Psychoneuroendocrinology</secondary-title></titles><periodical><full-title>Psychoneuroendocrinology</full-title></periodical><pages>557-569</pages><volume>35</volume><number>4</number><dates><year>2010</year></dates><isbn>0306-4530</isbn><urls></urls></record></Cite></EndNote>(27)The test stress not only disrupts the cognitive performance of first-year medical students, but also increases the psychological stress and the physiological variables of stress (heart rate, systolic and diastolic blood pressure). However, the relationship between the sympathetic–adrenal-medullary system parameters and salivary enzymes in different stress patterns, in particular psychological stress, such as the exam, is not clear. On the other hand, general tendency to anxiety and worry about failure during the written exam as psychological stress with release lysosomal enzymes has a deleterious effects on salivary gland cells ADDIN EN.CITE <EndNote><Cite><Author>Huxham</Author><Year>2012</Year><RecNum>32</RecNum><DisplayText>(28)</DisplayText><record><rec-number>32</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845211″>32</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Huxham, Mark</author><author>Campbell, Fiona</author><author>Westwood, Jenny</author></authors></contributors><titles><title>Oral versus written assessments: a test of student performance and attitudes</title><secondary-title>Assessment &amp; Evaluation in Higher Education</secondary-title></titles><periodical><full-title>Assessment &amp; Evaluation in Higher Education</full-title></periodical><pages>125-136</pages><volume>37</volume><number>1</number><dates><year>2012</year></dates><isbn>0260-2938</isbn><urls></urls></record></Cite></EndNote>(28)According to the results of this study ( Table 2), there was a positive and significant difference between salivary acid phosphatase, beta-glucuronidase and cathepsin activity under test conditions and two months before the exam (rest). The amount of activity of the acid phosphatase, ?-glucuronidase and catepsin during the test stress in saliva showed a significant increase. In general, this finding is consistent with previous studies PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5TaW5naDwvQXV0aG9yPjxZZWFyPjIwMTI8L1llYXI+PFJl
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ADDIN EN.CITE.DATA (1, 2, 29)on the effect of test stress on salivary enzymes and supports the effect of hypothesis psychological stressor (test) on salivary acid phosphate levels ADDIN EN.CITE <EndNote><Cite><Author>Kaufman</Author><Year>2000</Year><RecNum>36</RecNum><DisplayText>(5)</DisplayText><record><rec-number>36</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845395″>36</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Kaufman, Eliaz</author><author>Lamster, Ira B</author></authors></contributors><titles><title>Analysis of saliva for periodontal diagnosis: a review</title><secondary-title>Journal of clinical periodontology</secondary-title></titles><periodical><full-title>Journal of clinical periodontology</full-title></periodical><pages>453-465</pages><volume>27</volume><number>7</number><dates><year>2000</year></dates><isbn>0303-6979</isbn><urls></urls></record></Cite></EndNote>(5)Previous studies evaluation the stress effects on Mind Body Medicine Skills and changes in levels of cortisol, DHEA-S, testosterone, and IgA were measured in both groups (T2p)-post-intervention and (T2c)-control during the period of final examinations on first-year medical students. The consequence, showed that biological effects of the psychological stress of the exam cause the increase in salivary amylase, salivary cortisol, DHEA-S, testosterone, and IgA also salivary reduction ADDIN EN.CITE <EndNote><Cite><Author>Ng</Author><Year>2008</Year><RecNum>37</RecNum><DisplayText>(30, 31)</DisplayText><record><rec-number>37</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845525″>37</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Ng, Vivian</author><author>Koh, David</author><author>Wee, Andrew</author></authors></contributors><titles><title>Salivary alpha amylase levels under conditions of extreme examination stress</title><secondary-title>Psychological reports</secondary-title></titles><periodical><full-title>Psychological reports</full-title></periodical><pages>455-458</pages><volume>103</volume><number>2</number><dates><year>2008</year></dates><isbn>0033-2941</isbn><urls></urls></record></Cite><Cite><Author>Schoofs</Author><Year>2008</Year><RecNum>38</RecNum><record><rec-number>38</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845575″>38</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Schoofs, D</author><author>Hartmann, R</author><author>Wolf, OT</author></authors></contributors><titles><title>Neuroendocrine stress responses to an oral academic examination: no strong influence of sex, repeated participation and personality traits</title><secondary-title>Stress</secondary-title></titles><periodical><full-title>Stress</full-title></periodical><pages>52-61</pages><volume>11</volume><number>1</number><dates><year>2008</year></dates><isbn>1025-3890</isbn><urls></urls></record></Cite></EndNote>(30, 31)Recent studies on humans and animals indicate that stressful situation can cause increase enzymes lysosomal levels in biological fluids, including serum and saliva. Therefore, the relationship between stress and enzymes is determined by a positive association in higher levels of stress. Previous studies presented increased activity acid phosphatase, beta-glucuronidase and cathepsin as risk factors in pathogenesis cancer, alzheimer’s, periodontitis, Inflammation and swelling of the gums PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5Cb3NjaDwvQXV0aG9yPjxZZWFyPjE5OTY8L1llYXI+PFJl
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ADDIN EN.CITE.DATA (32, 33)However, the complex relationship between enzyme with stress requires further research on endocrine psychoanalysis ADDIN EN.CITE ;EndNote;;Cite;;Author;King;/Author;;Year;2005;/Year;;RecNum;41;/RecNum;;DisplayText;(34);/DisplayText;;record;;rec-number;41;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845742″;41;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;King, Jean A;/author;;author;Rosal, Milagros C;/author;;author;Ma, Yunsheng;/author;;author;Reed, George W;/author;;/authors;;/contributors;;titles;;title;Association of stress, hostility and plasma testosterone levels;/title;;secondary-title;Neuroendocrinology Letters;/secondary-title;;/titles;;periodical;;full-title;Neuroendocrinology Letters;/full-title;;/periodical;;pages;355-360;/pages;;volume;26;/volume;;number;4;/number;;dates;;year;2005;/year;;/dates;;isbn;0172-780X;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(34)In the present study, the sex of subjects did not affect the reaction of salivary enzymes to stress exame, and only stress exam factor was which increased the amount of salivary enzymes in both sexes. Another hypothesis in this study is the prediction of the relationship between levels of acid phosphatase, ?-glucuronidase and cathepsin under stressful conditions, with ruminant and five personality traits. In the ruminantion field , findings showe a positive correlation between rumination and stress enzymes. Often, evidence suggests that stress biomarker enzymes, as a potential mechanism, related to rumination with negative health outcomes. According to what we expected, rumination was associated with changing levels of salivary acid phosphatase, ?-glucuronidase and cathepsin under stress conditions. Previous research have found a relationship between rumination with salivary cortisol ADDIN EN.CITE ;EndNote;;Cite;;Author;Zoccola;/Author;;Year;2012;/Year;;RecNum;42;/RecNum;;DisplayText;(12);/DisplayText;;record;;rec-number;42;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845794″;42;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Zoccola, Peggy M;/author;;author;Dickerson, Sally S;/author;;/authors;;/contributors;;titles;;title;Assessing the relationship between rumination and cortisol: A review;/title;;secondary-title;Journal of Psychosomatic Research;/secondary-title;;/titles;;periodical;;full-title;Journal of Psychosomatic Research;/full-title;;/periodical;;pages;1-9;/pages;;volume;73;/volume;;number;1;/number;;dates;;year;2012;/year;;/dates;;isbn;0022-3999;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(12)amylase salivary and testosterone ADDIN EN.CITE ;EndNote;;Cite;;Author;Afrisham;/Author;;Year;2015;/Year;;RecNum;43;/RecNum;;DisplayText;(15);/DisplayText;;record;;rec-number;43;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845857″;43;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Afrisham, Reza;/author;;author;Sadegh-Nejadi, Sahar;/author;;author;SoliemaniFar, Omid;/author;;author;Abromand, Mohammad;/author;;author;Kooti, Wesam;/author;;author;Najjar Asl, Sedigheh;/author;;author;Khaneh Keshi, Ali;/author;;/authors;;/contributors;;titles;;title;Evaluating the salivary alpha-amylase level under psychological stress and its relationship with rumination and the five personality traits;/title;;secondary-title;Journal of Mazandaran University of Medical Sciences;/secondary-title;;/titles;;periodical;;full-title;Journal of Mazandaran University of Medical Sciences;/full-title;;/periodical;;pages;22-33;/pages;;volume;25;/volume;;number;126;/number;;dates;;year;2015;/year;;/dates;;urls;;/urls;;/record;;/Cite;;/EndNote;(15)Our findings indicated that the traits of neuroticism and agreeableness with acid phosphatase, neuroticism and extraversion with cathepsin and neuroticism with ?-glucuronidase have a positive communication with salivary enzymes under stress. These relationship show that the trait of neuroticism, agreeableness and extraversion can be associated with enhance salivary enzymes. The central element of neuroticism is the tendency to interpret the events as impregnated or injurious, which is linked to an increase in negative emotions. On the other hand, an increase in negative emotions in response to stress is associated with an increase in the level of salivary enzymes ADDIN EN.CITE ;EndNote;;Cite;;Author;Hauner;/Author;;Year;2008;/Year;;RecNum;44;/RecNum;;DisplayText;(35);/DisplayText;;record;;rec-number;44;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845904″;44;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Hauner, Katherina KY;/author;;author;Adam, Emma K;/author;;author;Mineka, Susan;/author;;author;Doane, Leah D;/author;;author;DeSantis, Amy S;/author;;author;Zinbarg, Richard;/author;;author;Craske, Michelle;/author;;author;Griffith, James W;/author;;/authors;;/contributors;;titles;;title;Neuroticism and introversion are associated with salivary cortisol patterns in adolescents;/title;;secondary-title;Psychoneuroendocrinology;/secondary-title;;/titles;;periodical;;full-title;Psychoneuroendocrinology;/full-title;;/periodical;;pages;1344-1356;/pages;;volume;33;/volume;;number;10;/number;;dates;;year;2008;/year;;/dates;;isbn;0306-4530;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(35)Slow heart rate and diastolic blood pressure responses to stress have been reported in hyper neuroticism patients ADDIN EN.CITE ;EndNote;;Cite;;Author;Oswald;/Author;;Year;2006;/Year;;RecNum;45;/RecNum;;DisplayText;(36);/DisplayText;;record;;rec-number;45;/rec-number;;foreign-keys;;key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533845943″;45;/key;;/foreign-keys;;ref-type name=”Journal Article”;17;/ref-type;;contributors;;authors;;author;Oswald, Lynn M;/author;;author;Zandi, Peter;/author;;author;Nestadt, Gerald;/author;;author;Potash, James B;/author;;author;Kalaydjian, Amanda E;/author;;author;Wand, Gary S;/author;;/authors;;/contributors;;titles;;title;Relationship between cortisol responses to stress and personality;/title;;secondary-title;Neuropsychopharmacology;/secondary-title;;/titles;;periodical;;full-title;Neuropsychopharmacology;/full-title;;/periodical;;pages;1583;/pages;;volume;31;/volume;;number;7;/number;;dates;;year;2006;/year;;/dates;;isbn;1740-634X;/isbn;;urls;;/urls;;/record;;/Cite;;/EndNote;(36)Comparing these correlations with the basal situation demonstrate that there is a relatively stable pattern of solidarity with a slight difference between neuroticism, agreeableness and extraversion with salivary enzymes. Personality traits with different biological reactions can be effective on a person’s perception of the stressful situation. Previous research on cognitive stress theory shown that personality trait influences the biological responses to stress. The role of stressors (such as the exam) in increasing the level of salivary enzymes can be due to factors such as novelty of conditions, uncertainty, variety, negative emotions and self-involvement of test conditions, age of subjects and individual differences in personality traits such as excitement will be dependent PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5XZWVrZXM8L0F1dGhvcj48WWVhcj4yMDA2PC9ZZWFyPjxS
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ADDIN EN.CITE.DATA (29, 37, 38)The regression analysis indicated that the traits of rumination, neuroticism, agreeableness and extraversion, have more roles in anticipate of the changes salivary enzymes under psychological stress ADDIN EN.CITE <EndNote><Cite><Author>Bibbey</Author><Year>2013</Year><RecNum>49</RecNum><DisplayText>(39)</DisplayText><record><rec-number>49</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533846107″>49</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Bibbey, Adam</author><author>Carroll, Douglas</author><author>Roseboom, Tessa J</author><author>Phillips, Anna C</author><author>de Rooij, Susanne R</author></authors></contributors><titles><title>Personality and physiological reactions to acute psychological stress</title><secondary-title>International journal of psychophysiology</secondary-title></titles><periodical><full-title>International journal of psychophysiology</full-title></periodical><pages>28-36</pages><volume>90</volume><number>1</number><dates><year>2013</year></dates><isbn>0167-8760</isbn><urls></urls></record></Cite></EndNote>(39)According to variable coefficients obtained, neuroticism, agreeableness and extraversion can anticipate a increase of salivary enzymes under psychological stress. The personality traits including neuroticism, agreeableness and extraversion can through the activation of neurochemical mechanisms, in prefrontal cortex of the brain, may increase or decrease the enzymes reactivity to stress. It may be the chief reason individual differences in biological reactivity to stress ADDIN EN.CITE <EndNote><Cite><Author>Terburg</Author><Year>2009</Year><RecNum>50</RecNum><DisplayText>(40, 41)</DisplayText><record><rec-number>50</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533846154″>50</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Terburg, David</author><author>Morgan, Barak</author><author>van Honk, Jack</author></authors></contributors><titles><title>The testosterone–cortisol ratio: A hormonal marker for proneness to social aggression</title><secondary-title>International journal of law and psychiatry</secondary-title></titles><periodical><full-title>International journal of law and psychiatry</full-title></periodical><pages>216-223</pages><volume>32</volume><number>4</number><dates><year>2009</year></dates><isbn>0160-2527</isbn><urls></urls></record></Cite><Cite><Author>Denson</Author><Year>2013</Year><RecNum>51</RecNum><record><rec-number>51</rec-number><foreign-keys><key app=”EN” db-id=”rw0avtfa1vd2zzewetqxz9d2w0wwzrwrftsr” timestamp=”1533846198″>51</key></foreign-keys><ref-type name=”Journal Article”>17</ref-type><contributors><authors><author>Denson, Thomas F</author><author>Ronay, Richard</author><author>von Hippel, William</author><author>Schira, Mark M</author></authors></contributors><titles><title>Endogenous testosterone and cortisol modulate neural responses during induced anger control</title><secondary-title>Social neuroscience</secondary-title></titles><periodical><full-title>Social neuroscience</full-title></periodical><pages>165-177</pages><volume>8</volume><number>2</number><dates><year>2013</year></dates><isbn>1747-0919</isbn><urls></urls></record></Cite></EndNote>(40, 41)In general, our results suggest that the variables associated with emotional control and five-factor personality model may illustrate individual differences in salivary enzymes reactivity to psychological stress. In fact, as mediating variables, personality trait and emotional control can increase or reduce the effects of stress. Although, few studies exist about the corrlation between psychological characteristics, especially five personality traits, with salivary acid phosphatase, ?-glucuronidase and cathepsin levels under psychological stress. Therefore, this study could be considered as a controversial and new study, but unlike experimental variables, many of the psychological variables are mutable. So, we need to perform further studies on psychological and personality factors and their interaction with changes salivary enzymes levels under stress condition.

Acknowledgments
This article has been adapted from a research project approved by the code of 92s86 and ethics code ajums. REC. in the Student Research Committee University of Ahvaz Jundishapur University of medical sciences; we appreciate the student research committee for preparation of grant. In addition, authors thank all medical students who participated in this study.

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