@2024 Afarand., IRAN
ISSN: 2251-8215 Sarem Journal of Reproductive Medicine 2018;2(3):93-97
ISSN: 2251-8215 Sarem Journal of Reproductive Medicine 2018;2(3):93-97
Comparison of Cytokines Levels Released from Th17 and Regulatory T Cells in Patients with Recurrent Spontaneous Abortions and Healthy Women
ARTICLE INFO
Article Type
Original ResearchAuthors
Roumandeh N. (1)Saremi A.T. (2)
Sanaye Naderi M. ()
Younesi B. (4)
Arasteh J. (5)
Zare A. (*)
() Sarem Fertility & Infertility Research Center (SAFIR), Sarem Women’s Hospital, Tehran, Iran
(*) “Sarem Fertility & Infertility Research Center (SAFIR)” and “Sarem Cell Research Center (SCRC)” , Shahid Beheshti Medical University, Tehran, Iran
(1) ”Sarem Cell Research Center (SCRC), Sarem Women’s Hospital, Tehran” and “Immunology Department, Medicine Faculty, Semnan University of Medical Sciences, Semnan, Iran
(2) “Sarem Fertility & Infertility Research Center (SAFIR)” and “Sarem Cell Research Center (SCRC)” , Sarem Women’s Hospital, Tehran, Iran
(4) Sarem Women’s Hospital, Tehran, Iran
(5) Biology Department, Basic Sciences Faculty, Central Tehran Branch, Islamic Azad University, Tehran, Iran
Correspondence
Article History
Received: February 9, 2017Accepted: June 27, 2017
ePublished: August 15, 2018
BRIEF TEXT
Successful pregnancy occurs as a result of maintaining the immune system of the mother to the fetus. Lack of maternal immunity system is associated with pregnancy problems and leads to abortion [1]. Evidence suggests that recurrent spontaneous abortion (RSA) is associated with inflammatory responses and immunological tolerance failure [2]. Three or more repeated abortions before the 20th week of pregnancy, is called recurrent spontaneous abortion.
The cause of this disorder is unknown in many cases, but there are several factors that can be attributed of which genetic abnormalities, hormonal disorders, abnormalities of the placenta and uterus, infections, and immunological factors can be named [3]. Among the immunologic factors, several mechanisms of environmental tolerance are involved in the preservation of the embryo, including the presence of T-regulatory cells (Treg) in the decidua and peripheral blood tissue during pregnancy [4]. On the other hand, the Th17 inflammatory cells secreting cytokine IL-17 are associated with red allograft rejection and deletion of tolerance. There is a growing evidence that regulated T cells and Th17 act as regulatory and executive cells in the establishment and maintenance of pregnancy, and impairment in their number and function may lead to failure of implantation and abnormalities of pregnancy [5]. During pregnancy, T-cells gradually increase in the first trimester and second trimester, and then decrease in the third trimester of pregnancy and after delivery [4, 6]. ... [7-14]. Th17 cells are closely related to Treg cells and, based on the findings, Treg cells are negatively associated with Th17 cells and serum IL-17 levels are positively related to Th17 cells and the ratio of Th17 to Treg cells [14]. Therefore, regulatory T cells inhibit the expression of IL-17, and the inhibition of Treg cells on Th17 cells may decrease in women with spontaneous abortion. In addition, it has been observed that cytokines secreted from Th17 cause rejection of allograft during pregnancy, while Treg-secreted cytokines are involved in preserving the tolerance of the fetus and may be effective in improving the outcome of pregnancy [14, 15]. In a study between fetal rejection and a reduction in the frequency of Treg cells, association has been observed [16]. The imbalance of Th17 cells to Treg in URSA is also involved [17].
Considering the role of cytokine balance in controlling the maternal immune response to embryo antigens during pregnancy, the study of cytokines involved in this process is important. The aim of this study was to evaluate the level of cytokines secreted from Th17 and T cells in patients with a history of unknown recurrent spontaneous abortion (URSA) compared to healthy women.
This is a case-control study.
This research was conducted on two groups of patients referred to Sarem Hospital in Tehran.
The case group was 30 patients with a history of three or more recurrent spontaneous abortions. Patients were included in the study after examining the status of karyotype, antiphospholipid antibodies, anti cardiolipin, ANA, anti-thyroid antibodies, microbial infections (TORCH, HBV, HIV, EBV, and CMV), anatomical abnormalities and hormonal disorders. Control subjects were 28 healthy women without a history of abortion and had at least one surviving child. Prior to sampling, the informed consent form approved by the Sarem Hospital's Ethics Committee was completed by each of the volunteer patients.
From each of the subjects in two groups, 2 cc of blood was taken and serum samples were stored in a freezer at -70 C for complete collection of the samples. To test the level of cytokines of Th17 cells including IL-17 and IL-21 and Treg cytokines including IL-10 and TGF-β in serum samples, ELISA technique was used with relevant kits (eBioscience, USA). Measuring the concentration of cytokines in patients’ samples according to the kit instructions was done in binary form. The sensitivity of the measurement kit for IL-17 and IL-21 cytokines was 8 and 0.01 Pico gram per milliliter respectively and it was 2, and 156.3 Pico gram per milliliterfor IL-10 and TGF-β cytokines respectively. Data were analyzed using SPSS 22 software. Normal distribution of data was investigated using Kolmogorov-Smirnov test. Non-parametric Mann-Whitney test was used to examine the significant difference between the groups studied considering the abnormal distribution of IL-21, IL-10, IL-17 and TGF-β data in the serum of patients and control group. Spearman correlation coefficient was used to determine the correlation between serum IL-17 concentration and TGF-β concentration in patients.
The mean age in the patient group was 31.03 ± 5.32 and in the control group was 33.71 ± 5.054 years. The concentration of IL-17 cytokines in serum samples was significantly higher in patients with URSA than in the control group (p <0.001). Also, serum TGF-β levels were significantly lower in the patients than in the control group (p = 0.001). Levels of IL-21 and IL-10 in the serum of patients were not significantly different in comparison with healthy women (p = 0.08, p = 0.02, respectively; Table 1).In addition, serum IL-17 concentration was positively and significantly correlated with TGF-β concentration in the patients (r = 0.554, p = 0.002, Figure 1).
... [18-21]. The results of this study showed a high level of IL-17 in serum of URSA patients compared to the control group, but the level of IL-21 in both groups was not significantly different. In a study by Sarshaki et al., It was also found that IL-17 levels in patients with this complication was higher compared to the control group [17]. In the similar study, the expression of IL-17 and IL-21 gene was investigated, based on which the expression of IL-17 gene in URSA patients had a higher level than normal pregnant women, while IL-21 gene expression in these patients had no statistically significant difference with the control group [22]. On the other hand, levels of TGF-β as a cytokine secreted from Treg cells showed a lower level in the serum of patients than the control group, which shows the role of Treg cell in maintaining pregnancy tolerance. In the present study, unlike most previous studies, there was no significant decrease in serum IL-10 levels in patients compared with healthy women. Nevertheless, these findings are consistent with the study by Yu et al., Which reported no significant differences in the level of IL-10 in serum URSA patients compared to healthy women [23]. Therefore, during pregnancy, the Th17 cytokine pattern may lead to fetal antigens and failure of the pregnancy's tolerance. Conversely, TGF-β can be an effective factor in maintaining this tolerance and the success of pregnancy. In this study, serum levels of IL-17 in URSA patients had a positive and significant correlation with TGF-β concentration in serum of these patients. Lee et al. also found that IL-17 + T cells in URSA women increased significantly compared to normal pregnant women, and there was a positive correlation between Th17 / Treg cell ratio and Th1 type cytokines [11].
Based on the results of this study and considering the role of pre-inflammatory cytokines of the Th17 type and the regulatory role of TGF-β, these cytokines can be used as a factor in assessing or even predicting the success or failure of pregnancy in URSA patients, which determines the significance of the Th17/Treg equilibrium more than before.
The level of IL-17 in women with a history of URSA is higher than that of healthy women and TGF-β levels is lower than those of healthy women, suggesting that Th17-immunity type and immunity regulation mediated by regularity T-cell are related to the pathogenesis of URSA.
The informed consent form approved by the morality committee of Sarem Hospital was completed by each of the volunteer patients.
TABLES and CHARTS
Show attach fileCITIATION LINKS
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[4]Pandey MK, Rani R, Agrawal S. An update in recurrent spontaneous abortion. Arch Gynecol Obstet. 2005;272(2):95-108.
[5]Beaman KD, Ntrivalas E, Mallers TM, Jaiswal MK, Kwak-Kim J, Gilman-Sachs A. Immune etiology of recurrent pregnancy loss and its diagnosis. Am J Reprod Immunol. 2012;67(4):319-25.
[6]Hanidziar D, Koulmanda M. Inflammation and the balance of Treg and Th17 cells in transplant rejection and tolerance. Curr Opin Organ Transplant. 2010;15(4):411-5.
[7]Sharma S. Natural killer cells and regulatory T cells in early pregnancy loss. Int J Dev Biol. 2014;58(2-4):219-29.
[8]Christiansen OB. Reproductive immunology. Mol Immunol. 2013;55(1):8-15.
[9]Kuon RJ, Strowitzki T, Sohn C, Daniel V, Toth B. Immune profiling in patients with recurrent miscarriage. J Reprod Immunol. 2015;108:136-41.
[10]Shoenfeld Y, Carp HJ, Molina V, Blank M, Cervera R, Balasch J, et al. Autoantibodies and prediction of reproductive failure. Am J Reprod Immunol. 2006;56(5-6):337-44.
[11]Cervera R, Balasch J. Autoimmunity and recurrent pregnancy losses. Clin Rev Allergy Immunol. 2010;39(3):148-52.
[12]Khonina NA, Broitman EV, Shevela EY, Pasman NM, Chernykh ER. Mixed lymphocyte reaction blocking factors (MLR-Bf) as potential biomarker for indication and efficacy of paternal lymphocyte immunization in recurrent spontaneous abortion. Arch Gynecol Obstet. 2013;288(4):933-7
[13]Bansal AS. Joining the immunological dots in recurrent miscarriage. Am J Reprod Immunol. 2010;64(5):307-15.
[14]Zhu LY, Chen X, Xu ZZ, Xu L, Mao T, Zhang H. Changes and clinical significance of peripheral blood helper T lymphocyte and natural killer (NK) cells in unexplained recurrent spontaneous abortion (URSA) patients after abortion and successful pregnancy. Clin Exp Obstet Gynecol. 2015;42(1):62-6.
[15]Yuan J, Li J, Huang SY, Sun X. Characterization of the subsets of human NKT-like cells and the expression of Th1/Th2 cytokines in patients with unexplained recurrent spontaneous abortion. J Reprod Immunol. 2015;110:81-8.
[16]Li X, Wang B, Li Y, Wang L, Zhao X, Zhou X, et al. The Th1/Th2/Th17/Treg paradigm induced by stachydrine hydrochloride reduces uterine bleeding in RU486-induced abortion mice. J Ethnopharmacol. 2013;145(1):241-53.
[17]Xu W, Roos A, Schlagwein N, Woltman AM, Daha MR, van Kooten C. IL-10-producing macrophages preferentially clear early apoptotic cells. Blood. 2006;107(12):4930-7.
[18]Nakashima A, Shima T, Inada K, Ito M, Saito S. The balance of the immune system between T cells and NK cells in miscarriage. Am J Reprod Immunol. 2012;67(4):304-10.
[19]Wang WJ, Hao CF, Qu QL, Wang X, Qiu LH, Lin QD. The deregulation of regulatory T cells on interleukin-17-producing T helper cells in patients with unexplained early recurrent miscarriage. Hum Reprod. 2010;25(10):2591-6.
[20]Sereshki N, Gharagozloo M, Ostadi V, Ghahiri A, Roghaei MA, Mehrabian F, et al. Variations in T-helper 17 and Regulatory T Cells during The Menstrual Cycle in Peripheral Blood of Women with Recurrent Spontaneous Abortion. Int J Fertil Steril. 2014;8(1):59-66.
[21]Arruvito L, Sotelo AI, Billordo A, Fainboim L. A physiological role for inducible FOXP3(+) Treg cells. Lessons from women with reproductive failure. Clin Immunol. 2010;136(3):432-41.
[22]Lee SK, Kim JY, Hur SE, Kim CJ, Na BJ, Lee M, et al. An imbalance in interleukin-17-producing T and Foxp3(+) regulatory T cells in women with idiopathic recurrent pregnancy loss. Hum Reprod. 2011;26(11):2964-71.
[23]Kessel A, Ammuri H, Peri R, Pavlotzky ER, Blank M, Shoenfeld Y, et al. Intravenous immunoglobulin therapy affects T regulatory cells by increasing their suppressive function. J Immunol. 2007;179(8):5571-5.
[24]Gomaa MF, Elkholy AG, El-Said MM, Abdel-Salam NE. Combined oral prednisolone and heparin versus heparin: the effect on peripheral NK cells and clinical outcome in patients with unexplained recurrent miscarriage. A double-blind placebo randomized controlled trial. Arch Gynecol Obstet. 2014; 290(4): 757–62.
[25]Areia AL, Fonseca E, Areia M, Moura P. Low-molecular-weight heparin plus aspirin versus aspirin alone in pregnant women with hereditary thrombophilia to improve live birth rate: Meta-analysis of randomized controlled trials. Arch Gynecol Obstet. 2016 Jan;293(1):81-6.
[26]Yuksel H, Kayatas S, Boza AT, Api M, Ertekin AA, Cam C. Low molecular weight heparin use in unexplained recurrent miscarriage. Pak J Med Sci. 2014; 30(6): 1232–7.
[27]Fawzy M, Shokeir T, El-Tatongy M, Warda O, El-Refaiey AA, Mosbah A. Treatment options and pregnancy outcome in women with idiopathic recurrent miscarriage: A randomized placebo-controlled study. Arch Gynecol Obstet. 2008;278(1):33-8.
[28]Kaandorp SP, Goddijn M, van der Post JA, Hutten BA, Verhoeve HR, Hamulyak K, et al. Aspirin plus heparin or aspirin alone in women with recurrent miscarriage. N Engl J Med. 2010;362(17):1586-96.
[29]Hussain M, El-Hakim S, Cahill DJ. Progesterone supplementation in women with otherwise unexplained recurrent miscarriages. J Hum Reprod Sci. 2012;5(3):248-51.
[30]Bansal AS, Bajardeen B, Thum MY. The basis and value of currently used immunomodulatory therapies in recurrent miscarriage. J Reprod Immunol. 2012;93(1):41-51.
[31]Practice Committee of the American Society for Reproductive Medicine. Intravenous immunoglobulin (IVIG) and recurrent spontaneous pregnancy loss. Fertil Steril. 2004;82 Suppl 1:s199-200.
[32]Egerup P, Lindschou J, Gluud C, Christiansen OB. The effects of immunotherapy with intravenous immunoglobulins versus no intervention, placebo, or usual care in patients with recurrent miscarriages: A protocol for a systematic review with meta-analyses, trial sequential analyses, and individual patient data meta-analyses of randomised clinical trials. Syst Rev. 2014;3:89.
[33]Takeshita T. Diagnosis and treatment of recurrent miscarriage associated with immunologic disorders: Is paternal lymphocyte immunization a relic of the past?. J Nippon Med Sch. 2004;71(5):308-13.
[34]Mowbray JF, Gibbings C, Liddell H, Reginald PW, Underwood JL, Beard RW. Controlled trial of treatment of recurrent spontaneous abortion by immunisation with paternal cells. Lancet. 1985;1(8435):941-3.
[35]Pandey MK, Thakur S, Agrawal S. Lymphocyte immunotherapy and its probable mechanism in the maintenance of pregnancy in women with recurrent spontaneous abortion. Arch Gynecol Obstet. 2004;269(3):161-72.
[36]Kano T, Mori T, Furudono M, Ishikawa H, Watanabe H, Kikkawa E, et al. Human leukocyte antigen may predict outcome of primary recurrent spontaneous abortion treated with paternal lymphocyte alloimmunization therapy. Am J Reprod Immunol. 2007;58(4):383-7.
[37]Ito K, Tanaka T, Tsutsumi N, Obata F, Kashiwagi N. Possible mechanisms of immunotherapy for maintaining pregnancy in recurrent spontaneous aborters: Analysis of anti-idiotypic antibodies directed against autologous T-cell receptors. Hum Reprod. 1999;14(3):650-5.
[2]Hogge WA, Byrnes AL, Lanasa MC, Surti U. The clinical use of karyotyping spontaneous abortions. Am J Obstet Gynecol. 2003;189(2):397-400.
[3]Christiansen OB, Nielsen HS, Kolte AM. Future directions of failed implantation and recurrent miscarriage research. Reprod Biomed Online. 2006;13(1):71-83.
[4]Pandey MK, Rani R, Agrawal S. An update in recurrent spontaneous abortion. Arch Gynecol Obstet. 2005;272(2):95-108.
[5]Beaman KD, Ntrivalas E, Mallers TM, Jaiswal MK, Kwak-Kim J, Gilman-Sachs A. Immune etiology of recurrent pregnancy loss and its diagnosis. Am J Reprod Immunol. 2012;67(4):319-25.
[6]Hanidziar D, Koulmanda M. Inflammation and the balance of Treg and Th17 cells in transplant rejection and tolerance. Curr Opin Organ Transplant. 2010;15(4):411-5.
[7]Sharma S. Natural killer cells and regulatory T cells in early pregnancy loss. Int J Dev Biol. 2014;58(2-4):219-29.
[8]Christiansen OB. Reproductive immunology. Mol Immunol. 2013;55(1):8-15.
[9]Kuon RJ, Strowitzki T, Sohn C, Daniel V, Toth B. Immune profiling in patients with recurrent miscarriage. J Reprod Immunol. 2015;108:136-41.
[10]Shoenfeld Y, Carp HJ, Molina V, Blank M, Cervera R, Balasch J, et al. Autoantibodies and prediction of reproductive failure. Am J Reprod Immunol. 2006;56(5-6):337-44.
[11]Cervera R, Balasch J. Autoimmunity and recurrent pregnancy losses. Clin Rev Allergy Immunol. 2010;39(3):148-52.
[12]Khonina NA, Broitman EV, Shevela EY, Pasman NM, Chernykh ER. Mixed lymphocyte reaction blocking factors (MLR-Bf) as potential biomarker for indication and efficacy of paternal lymphocyte immunization in recurrent spontaneous abortion. Arch Gynecol Obstet. 2013;288(4):933-7
[13]Bansal AS. Joining the immunological dots in recurrent miscarriage. Am J Reprod Immunol. 2010;64(5):307-15.
[14]Zhu LY, Chen X, Xu ZZ, Xu L, Mao T, Zhang H. Changes and clinical significance of peripheral blood helper T lymphocyte and natural killer (NK) cells in unexplained recurrent spontaneous abortion (URSA) patients after abortion and successful pregnancy. Clin Exp Obstet Gynecol. 2015;42(1):62-6.
[15]Yuan J, Li J, Huang SY, Sun X. Characterization of the subsets of human NKT-like cells and the expression of Th1/Th2 cytokines in patients with unexplained recurrent spontaneous abortion. J Reprod Immunol. 2015;110:81-8.
[16]Li X, Wang B, Li Y, Wang L, Zhao X, Zhou X, et al. The Th1/Th2/Th17/Treg paradigm induced by stachydrine hydrochloride reduces uterine bleeding in RU486-induced abortion mice. J Ethnopharmacol. 2013;145(1):241-53.
[17]Xu W, Roos A, Schlagwein N, Woltman AM, Daha MR, van Kooten C. IL-10-producing macrophages preferentially clear early apoptotic cells. Blood. 2006;107(12):4930-7.
[18]Nakashima A, Shima T, Inada K, Ito M, Saito S. The balance of the immune system between T cells and NK cells in miscarriage. Am J Reprod Immunol. 2012;67(4):304-10.
[19]Wang WJ, Hao CF, Qu QL, Wang X, Qiu LH, Lin QD. The deregulation of regulatory T cells on interleukin-17-producing T helper cells in patients with unexplained early recurrent miscarriage. Hum Reprod. 2010;25(10):2591-6.
[20]Sereshki N, Gharagozloo M, Ostadi V, Ghahiri A, Roghaei MA, Mehrabian F, et al. Variations in T-helper 17 and Regulatory T Cells during The Menstrual Cycle in Peripheral Blood of Women with Recurrent Spontaneous Abortion. Int J Fertil Steril. 2014;8(1):59-66.
[21]Arruvito L, Sotelo AI, Billordo A, Fainboim L. A physiological role for inducible FOXP3(+) Treg cells. Lessons from women with reproductive failure. Clin Immunol. 2010;136(3):432-41.
[22]Lee SK, Kim JY, Hur SE, Kim CJ, Na BJ, Lee M, et al. An imbalance in interleukin-17-producing T and Foxp3(+) regulatory T cells in women with idiopathic recurrent pregnancy loss. Hum Reprod. 2011;26(11):2964-71.
[23]Kessel A, Ammuri H, Peri R, Pavlotzky ER, Blank M, Shoenfeld Y, et al. Intravenous immunoglobulin therapy affects T regulatory cells by increasing their suppressive function. J Immunol. 2007;179(8):5571-5.
[24]Gomaa MF, Elkholy AG, El-Said MM, Abdel-Salam NE. Combined oral prednisolone and heparin versus heparin: the effect on peripheral NK cells and clinical outcome in patients with unexplained recurrent miscarriage. A double-blind placebo randomized controlled trial. Arch Gynecol Obstet. 2014; 290(4): 757–62.
[25]Areia AL, Fonseca E, Areia M, Moura P. Low-molecular-weight heparin plus aspirin versus aspirin alone in pregnant women with hereditary thrombophilia to improve live birth rate: Meta-analysis of randomized controlled trials. Arch Gynecol Obstet. 2016 Jan;293(1):81-6.
[26]Yuksel H, Kayatas S, Boza AT, Api M, Ertekin AA, Cam C. Low molecular weight heparin use in unexplained recurrent miscarriage. Pak J Med Sci. 2014; 30(6): 1232–7.
[27]Fawzy M, Shokeir T, El-Tatongy M, Warda O, El-Refaiey AA, Mosbah A. Treatment options and pregnancy outcome in women with idiopathic recurrent miscarriage: A randomized placebo-controlled study. Arch Gynecol Obstet. 2008;278(1):33-8.
[28]Kaandorp SP, Goddijn M, van der Post JA, Hutten BA, Verhoeve HR, Hamulyak K, et al. Aspirin plus heparin or aspirin alone in women with recurrent miscarriage. N Engl J Med. 2010;362(17):1586-96.
[29]Hussain M, El-Hakim S, Cahill DJ. Progesterone supplementation in women with otherwise unexplained recurrent miscarriages. J Hum Reprod Sci. 2012;5(3):248-51.
[30]Bansal AS, Bajardeen B, Thum MY. The basis and value of currently used immunomodulatory therapies in recurrent miscarriage. J Reprod Immunol. 2012;93(1):41-51.
[31]Practice Committee of the American Society for Reproductive Medicine. Intravenous immunoglobulin (IVIG) and recurrent spontaneous pregnancy loss. Fertil Steril. 2004;82 Suppl 1:s199-200.
[32]Egerup P, Lindschou J, Gluud C, Christiansen OB. The effects of immunotherapy with intravenous immunoglobulins versus no intervention, placebo, or usual care in patients with recurrent miscarriages: A protocol for a systematic review with meta-analyses, trial sequential analyses, and individual patient data meta-analyses of randomised clinical trials. Syst Rev. 2014;3:89.
[33]Takeshita T. Diagnosis and treatment of recurrent miscarriage associated with immunologic disorders: Is paternal lymphocyte immunization a relic of the past?. J Nippon Med Sch. 2004;71(5):308-13.
[34]Mowbray JF, Gibbings C, Liddell H, Reginald PW, Underwood JL, Beard RW. Controlled trial of treatment of recurrent spontaneous abortion by immunisation with paternal cells. Lancet. 1985;1(8435):941-3.
[35]Pandey MK, Thakur S, Agrawal S. Lymphocyte immunotherapy and its probable mechanism in the maintenance of pregnancy in women with recurrent spontaneous abortion. Arch Gynecol Obstet. 2004;269(3):161-72.
[36]Kano T, Mori T, Furudono M, Ishikawa H, Watanabe H, Kikkawa E, et al. Human leukocyte antigen may predict outcome of primary recurrent spontaneous abortion treated with paternal lymphocyte alloimmunization therapy. Am J Reprod Immunol. 2007;58(4):383-7.
[37]Ito K, Tanaka T, Tsutsumi N, Obata F, Kashiwagi N. Possible mechanisms of immunotherapy for maintaining pregnancy in recurrent spontaneous aborters: Analysis of anti-idiotypic antibodies directed against autologous T-cell receptors. Hum Reprod. 1999;14(3):650-5.