- Email: [email protected]
Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers
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MODEL
Pediatrics and Neonatology (2014) xx, 1e4
Available online at www.sciencedirect.com
ScienceDirect journal homepage: http://www.pediatr-neonatol.com
ORIGINAL ARTICLE
Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers Naci Topalo glu a, S‚ule Yıldırım a,*, Mustafa Tekin a, ¨ zdemir a, ¨tu ¨ncu ¨ler c, Cem O Nazan Kaymaz b, Funda Tu Emine Cos‚ar b a
C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Pediatrics, C ¸ anakkale, Turkey C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Obstetrics and Gynecology, C ¸ anakkale, Turkey c C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Biochemistry, C ¸ anakkale, Turkey b
Received Oct 11, 2013; received in revised form Dec 30, 2013; accepted Feb 5, 2014
Key Words gestational diabetes; oxidative stress; mean platelet volume; ischemia modified albumin; cord blood
Background: Gestational diabetes mellitus (GDM) is a risk for the health of both the pregnant women and her infant. Its unfavorable effects start in utero and continue after birth. It is known that GDM increases oxidative stress and decreases antioxidant enzyme activities. In this study we aimed to investigate cord blood mean platelet volume (MPV) and ischemia-modified albumin (IMA) levels of infants of diabetic mothers (IDM). Methods: Twenty-nine pregnant women with GDM between 37 and 41 gestational weeks who gave birth by spontaneous vaginal delivery were enrolled as study participants together with 20 healthy pregnant women as a control group. Weight, length, and head circumference of babies were measured by the same standard tape immediately after birth. Five milliliters of umbilical venous blood were obtained to study MPV and IMA levels. Results: There was statistically significant difference in levels of MPV (p Z 0.037) and IMA (p < 0.001) between groups. They increased in IDM compared with their healthy peers. Conclusions: Evaluation of MPV and IMA together is useful for representing the potential oxidative stress of IDM. Copyright ª 2014, Taiwan Pediatric Association. Published by Elsevier Taiwan LLC. All rights reserved.
¨ ni* Corresponding author. C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Pediatrics, C ¸ anakkale Onsekiz Mart U versitesi Tıp Faku ¸ anakkale, Turkey. ¨ltesi Hastanesi, Cumhuriyet Mahallesi Sahil Yolu Number 5, Kepez, C E-mail address: [email protected] (S‚. Yıldırım). http://dx.doi.org/10.1016/j.pedneo.2014.02.002 1875-9572/Copyright ª 2014, Taiwan Pediatric Association. Published by Elsevier Taiwan LLC. All rights reserved. Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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N. Topalo glu et al
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1. Introduction Prevalence of gestational diabetes mellitus (GDM) is increasing worldwide and it is reported to range from 1% to 14% in the literature.1 Race, increasing age, and obesity are the risk factors. Early diagnosis and adequate treatment is important for prevention of complications. While preeclampsia, polyhydramnios, fetal macrosomia, and operative delivery are complications that are seen in GDM pregnant women, hypoglycemia, hypocalcemia, hyperbilirubinemia, and polycytemia are some of the complications seen in the infants of diabetic mothers (IDM). Besides an increment in perinatal mortality, the risk of developing type II diabetes mellitus in the following 10 years increases by 20e30%. Obesity and impaired glucose tolerance ratios increase in the childhood period of IDM.2 Literature about the oxidative stress increasing effect of GDM and decreased antioxidant enzyme capacity in those pregnant women is limited.3 Ischemia modified albumin (IMA) is a biomarker for acute ischemia that is approved by the US Food and Drug Administration. When exposed to ischemic conditions, the N-terminus of albumin is damaged, which makes it unable to bind metals and capable of being measured by an albumin cobalt-binding test. Because its levels in the blood increase within minutes of the onset of ischemia and return to normal within 6e12 hours, IMA has been implicated in the detection of acute ischemia prior to necrosis.4e6 Pregnancy generates a stressful condition in the woman that is related to increased oxidative stress.7 Gugliucci et al demonstrated a marked difference of IMA levels between pregnant and nonpregnant women. IMA levels also increase in normal vaginal delivery.8e10 There is insufficient literature investigating the correlation between GDM and IMA levels. Ma et al found that high levels of IMA and blood glucose levels in GDM pregnant women significantly decreased to normal levels after 6 weeks of regular treatment. 11 The mean platelet volume (MPV) is a useful marker to determine platelet morphology. A normal range of cord blood MPV levels in healthy newborns has been defined.12 Kipper et al12 also revealed the alterations in thrombocyte and IMA level in mature and premature infants. Diabetes also changes thrombocyte function and morphology. Increased MPV is a direct sign of thrombocyte synthesis and activation. MPV increases as a result of thrombocyte synthesis. Sak et al showed a clear increase in the MPV of GDM pregnant women compared with the control group.13 To our knowledge, this is the first study to investigate the cord blood IMA and MPV levels in the same group of diabetic patients. In our study, our aim is to investigate these parameters as a sign of oxidative stress in the IDM.
2. Materials and methods We performed a caseecontrol study from June 2012 to July 2013 at the Departments of Obstetrics and Gynecology and Pediatrics at C ¸ anakkale Onsekiz Mart University Faculty of Medicine. The approval of the ethics committee of our university was obtained. Every patient included in the study signed an informed consent form before participating in the study. We enrolled 29 GDM pregnant women between 37 and 41 gestational weeks who gave birth by spontaneous
vaginal delivery together with 20 healthy pregnant women as a control group. Pregnant women who did not want to enroll in the study, gave birth by cesarean delivery, those who had multiple pregnancies, those with chronic diseases and fetal anomalies, those using chronic medications, and those who smoked were excluded from the study. Postpartum 1st minute and 5th minute APGAR scores were recorded. Babies were weighed without clothes immediately after birth. The length of the baby and head circumference were measured using the same standard tape. Five milliliters of umbilical venous blood were obtained just after delivery into dry tubes with separating gel. Blood was centrifuged at 800 g at 4 C for 15 minutes and the separated serum was immediately analyzed or frozen at 80 C until use. Spectrophotometric measurements were made in a Beckman DU 640 spectrophotometer (Beckman Coulter Inc, Fullerton, CA, USA). IMA was measured by the decrease in cobalt 2þ binding as previously described.4 We introduced minor modifications in the method to adapt it to a 96-well plate reader. Briefly, we added 100 mL of patient serum to 25 mL of a solution of 1 g/L cobalt chloride and a blank for each serum was prepared similarly; þ25 mL of a 9.0 g/L solution of NaCl. After agitation and 10 minutes’ incubation at 25 C, dithiothreitol (25 mL of a 1.5 g/L solution) was then added. After mixing and 2 minutes’ incubation, the absorbance of assay mixtures was read at 470 nm, zeroing with the individual blank. IMA values are expressed in absorbance units (AU). Platelet count and MPV determinations were performed based on the Coulter Counter model LH (Coulter Electronics, Hialeah, FL, USA). Data were recorded using the SPSS 15.0 statistical software package (SPSS Inc., Chicago, IL, USA). Descriptive data are shown as mean standard deviation. ManneWhitney U test was applied for comparisons. A p value < 0.05 was considered significant.
3. Results Table 1 shows the cord blood levels of IMA and MPV in the IDM and control groups. The serum IMA levels were significantly higher in the IDM group compared with the control group (p < 0.001; Figure 1). The mean IMA value was 912.20 92.41 AU in IDM group and 715.60 150.35 AU in the control group. There was also a statistically significant difference between groups in MPV values. The serum MPV was significantly higher in the IDM group (p Z 0.037; Figure 2). In the IDM group, the mean serum MPV was 8.11 0.95 fL and in control group it was 7.55 0.72 fL. Table 1 Cord blood mean platelet volume (MPV) and ischemia modified albumin (IMA) levels of participants. GDM pregnant women
Healthy controls p
MPV, fL 8.11 0.95 7.55 0.72 0.037 Mean SD IMA, absorbance 912.20 92.41 715.60 150.35 <0.001 unit Mean SD Data are presented as the mean standard deviation. GDM Z gestational diabetes mellitus.
Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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MPV and IMA in infants of diabetic mothers
3 Table 2
Characteristics of case and control groups. Case
Control
Gestational week 39.1 0.9 39.0 1.2 Length, cm 49.5 1.6 49.7 1.9 Head circumference, cm 34.5 1.7 34.4 1.34 Birth weight, g 3164 551 3212 509 212.5 68.4 221.9 85.4 Platelet count 1012 Age of mother, y 31.4 5.3 26.8 5.0
p 0.838 0.657 0.967 0.799 0.640 0.004
Data are presented as the mean standard deviation.
Figure 1 Cord blood ischemia-modified albumin (IMA) levels of infants of diabetic mothers and healthy controls. The difference is statistically significant (p < 0.001).
Table 2 represents the demographic characteristics of the newborns and pregnant women in both groups. There was no statistically significant difference between the clinical characteristics of the groups except in maternal age (p > 0.05). The difference in maternal age between groups did not affect the IMA level or MPV.
4. Discussion We found a significant elevation in the cord blood level of IMA and MPV in IDM as evidence of danger to health. Pregnancy generates a hypoxic condition in experimental models. Antioxidative defensive mechanisms change during pregnancy. IMA is used to predict hypoxic conditions, so it may be used in pregnancy to show hypoxemia. Some studies have reported increased IMA levels dependent on oxidative stress triggered by pregnancy.8,9
Figure 2 Cord blood mean platelet volume (MPV) of infants of diabetic mothers and healthy controls. The difference is statistically significant (p Z 0.037).
There is evidence that pregnancy is a physiologically increased stress condition and in complicated pregnancies ¨ zdemir et al IMA levels increase more than normal ones. O found significantly higher levels of IMA in pregnant women with habitual abortions.14 This finding may suggest that an abnormally high hypoxic intrauterine environment may be associated with abnormal placental development that contributes to early miscarriage. Pregnant women carrying fetuses with neural tube defects had higher levels of IMA ¨ stu when compared to normal pregnant women.15 U ¨n et al found that moderate and severe preeclamptic pregnant patients had higher IMA levels than healthy pregnant women and IMA levels positively correlated with the severity of preeclampsia.16 In that study they also showed that healthy and preeclamptic pregnant women had higher IMA levels than nonpregnant women. GDM is a risk for the health of pregnant women and their infants. Women with risk factors should be screened and early diagnosis should be made for prevention of complications. The best test for the diagnosis is oral glucose tolerance test; in our study, all the patients were positive for this test. Oxidative stress had positive correlation with increased blood glucose levels, protein oxidation and lipid peroxidase concentration capacity in GDM pregnant women.17 Ma et al found high levels of IMA in GDM pregnant women compared with healthy ones and they attributed this result to high blood glucose levels.11 In our study, we found higher levels of IMA in cord blood of IDM than the controls. GDM is a systemic disease that affects both mother and fetus.18,19 The platelet count is slightly lower in pregnant than in nonpregnant women.20 Platelet levels also decrease while the duration of pregnancy increases.21 Platelet volume is a marker of platelet activation and function and is measured using the MPV.18,19 MPV values have been found to be higher in diabetic patients when compared to normal controls. When Sharpe and Trinick compared the MPV of GDM and healthy pregnant women, they found significantly higher MPV in GDM pregnant women.22 Kharb23 claimed that low insulin sensitivity is the cause of the oxidative stress and leads to free radical production. Because GDM markedly triggers oxidative stress and MPV directly shows the response of the thrombocytes to the stress, MPV may be used as a marker for oxidative stress. Patients with diabetes have increased platelet activation compared to nondiabetic individuals.19,24,25 MPV values can serve as an effective marker of blood glucose levels.18,26 Recently, Bozkurt et al19 claimed that GDM patients had higher MPV values than normal controls and that patients with high MPV values had low platelet counts. It
Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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4 has been reported that platelet survival is shorter in diabetic patients.19,26 MPV increment also showed in metabolic syndrome,27 myocardial infarction,28 and stroke29 cases. S‚ahin et al30 found that severe preeclamptic pregnant women had higher MPVs than mild preeclamptic and healthy pregnant women. Large thrombocytes are more active, produce more prothrombotic factors and adhere easily.31 We did not find statistically significant difference between the groups in thrombocyte counts. MPVs are statistically higher in IDM in our study and this finding is supported by the literature. In conclusion, evaluation of MPV and IMA is useful for representing the potential oxidative stress of IDM.
Conflicts of interest Financial disclosure statements have been obtained, and no conflicts of interest have been reported by the authors or by any individuals in control of the content of this article.
References 1. Jovanovic L, Pettitt DJ. Gestational diabetes mellitus. JAMA 2001;286:2516e8. 2. Ogata ES. Perinatal morbidity in offspring of diabetic mothers. Diabetes Rev 1995;3:652e7. 3. Makedou K, Kourtis A, Gkiomisi A, Toulis KA, Mouzaki M, Anastasilakis AD, et al. Oxidized low-density lipoprotein and adiponectin levels in pregnancy. Gynecol Endocrinol 2011;27: 1070e3. 4. Bar-Or D, Lau E, Winkler JV. A novel assay for cobalt-albumin binding and its potential as a marker for myocardial ischemia-a preliminary report. J Emerg Med 2000;19:311e5. 5. Hjortshøj S, Kristensen SR, Ravkilde J. Diagnostic value of ischemia modified albumin in patients with suspected acute coronary syndrome. Am J Emerg Med 2010;28:170e6. 6. Kanko M, Yavuz S, Duman C, Hosten T, Oner E, Berki T. Ischemia-modified albumin use as a prognostic factor in coronary bypass surgery. J Cardiothorac Surg 2012;7:3. 7. Prefumo F, Gaze DC, Papageorghiou AT, Collinson PO, Thilaganathan B. First trimester maternal serum ischaemiamodified albumin: a marker of hypoxia-ischaemia-driven early trophoblast development. Hum Reprod 2007;22: 2029e32. 8. van Rijn BB, Franx A, Sikkema JM, van Rijn HJ, Bruinse HW, Voorbij HA. Ischemia modified albumin in normal pregnancy and preeclampsia. Hypertens Pregnancy 2008;27:159e67. 9. Guven S, Alver A, Mentese A, Ilhan FC, Calapoglu M, Unsal MA. The novel ischemia marker ‘ischemia-modified albumin’ is increased in normal pregnancies. Acta Obstet Gynecol Scand 2009;88:479e82. 10. Gugliucci A, Hermo R, Monroy C, Numaguchi M, Kimura S. Ischemia-modified albumin levels in cord blood: A case-control study in uncomplicated and complicated deliveries. Clin Chim Acta 2005;362:155e60. 11. Ma SG, Yu WN, Jin Y, Hong B, Hu W. Evaluation of serum ischemia-modified albumin levels in pregnant women with and without gestational diabetes mellitus. Gynecol Endocrinol 2012;28:837e40.
N. Topalo glu et al 12. Kipper SL, Sieger L. Whole blood platelet volumes in newborn infants. J Pediatr 1982;101:763e6. ¨ zler A, Evsen MS, Turgut A, Sak S, et al. 13. Sak ME, Soydinc ¸ HE, O Platelet profile in patients with gestational diabetes: a retrospective study. J Turk Ger Gynecol Assoc 2012;13:223e6. ¨ zdemir S, Kıyıcı A, Balci O, Go 14. O ¸ ic ¸ekler H, C ¸ elik C ¸. ¨ktepe H, C Assessment of ischemia-modified albumin level in patients with recurrent pregnancy loss during the first trimester. Eur J Obstet Gynecol Reprod Biol 2011;155:209e12. 15. Gu ¨zelmansur I, Ustu ¨n Y, Engin-Ustu ¨n Y, Oztu ¨rk O, Yaman H. Serum ischemia modified albumin levels in pregnancies with neural tube defects. J Reprod Med 2012;57:49e52. 16. Ustu ¨n Y, Engin-Ustu ¨n Y, Oztu ¨rk O, Alanbay I, Yaman H. Ischemia-modified albumin as an oxidative stress marker in preeclampsia. J Matern Fetal Neonatal Med 2011;24:418e21. 17. Boisvert MR, Koski KG, Skinner CD. Increased oxidative modifications of amniotic fluid albumin in pregnancies associated with gestational diabetes mellitus. Anal Chem 2010;82: 1133e7. 18. Hekimsoy Z, Payzin B, Ornek T, Kandogan G. Mean platelet volume in Type 2 diabetic patients. J Diabetes Complications 2004;18:173e6. 19. Bozkurt N, Yilmaz E, Biri A, Taner Z, Himmetoglu O. The mean platelet volume in gestational diabetes. J Thromb Thrombolysis 2006;22:51e4. 20. Abbassi-Ghanavati M, Greer LG, Cunningham FG. Pregnancy and laboratory studies: a reference table for clinicians. Obstet Gynecol 2009;114:1326e31. 21. Akingbola TS, Adewole IF, Adesina OA, Afolabi KA, Fehintola FA, Bamgboye EA, et al. Haematological profile of healthy pregnant women in Ibadan, south-western Nigeria. J Obstet Gynaecol 2006;26:763e9. 22. Sharpe PC, Trinick T. Mean platelet volume in diabetes mellitus. Q J Med 1993;86:739e42. 23. Kharb S. Lipid peroxidation in pregnancy with preeclampsia and diabetes. Gynecol Obstet Invest 2000;50:113e6. 24. Erikc ¸i AA, Muhc ¸u M, Du ¨ndar O, Oztu ¨rk A. Could mean platelet volume be a predictive marker for gestational diabetes mellitus? Hematology 2008;13:46e8. 25. Dundar O, Yoruk P, Tutuncu L, Erikci AA, Muhcu M, Ergur AR, et al. Longitudinal study of platelet size changes in gestation and predictive power of elevated MPV in development of preeclampsia. Prenat Diagn 2008;28:1052e6. 26. Coban E, Bostan F, Ozdogan M. The mean platelet volume in subjects with impaired fasting glucose. Platelets 2006;17: 67e9. Erratum in: Platelets 2006;17:350. 27. Tavil Y, Sen N, Yazici HU, Hizal F, Abaci A, Cengel A. Mean platelet volume in patients with metabolic syndrome and its relationship with coronary artery disease. Thromb Res 2007; 120:245e50. 28. Khandekar MM, Khurana AS, Deshmukh SD, Kakrani AL, Katdare AD, Inamdar AK. Platelet volume indices in patients with coronary artery disease and acute myocardial infarction: an Indian scenario. J Clin Pathol 2006;59:146e9. 29. O’Malley T, Langhorne P, Elton RA, Stewart C. Platelet size in stroke patients. Stroke 1995;26:995e9. 30. S‚ahin HG, S‚ahin HA, Zeteroglu S‚, Kamacı M, Surucu R, Guvercinci M. Mean platelet volume levels in preeclampsia and eclampsia. Turkiye Klinikleri J Gynecol Obst 2002;12:126e9. 31. Bath PM, Butterworth RJ. Platelet size: measurement, physiology and vascular disease. Blood Coagul Fibrinolysis 1996;7: 157e61.
Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
MODEL
Pediatrics and Neonatology (2014) xx, 1e4
Available online at www.sciencedirect.com
ScienceDirect journal homepage: http://www.pediatr-neonatol.com
ORIGINAL ARTICLE
Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers Naci Topalo glu a, S‚ule Yıldırım a,*, Mustafa Tekin a, ¨ zdemir a, ¨tu ¨ncu ¨ler c, Cem O Nazan Kaymaz b, Funda Tu Emine Cos‚ar b a
C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Pediatrics, C ¸ anakkale, Turkey C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Obstetrics and Gynecology, C ¸ anakkale, Turkey c C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Biochemistry, C ¸ anakkale, Turkey b
Received Oct 11, 2013; received in revised form Dec 30, 2013; accepted Feb 5, 2014
Key Words gestational diabetes; oxidative stress; mean platelet volume; ischemia modified albumin; cord blood
Background: Gestational diabetes mellitus (GDM) is a risk for the health of both the pregnant women and her infant. Its unfavorable effects start in utero and continue after birth. It is known that GDM increases oxidative stress and decreases antioxidant enzyme activities. In this study we aimed to investigate cord blood mean platelet volume (MPV) and ischemia-modified albumin (IMA) levels of infants of diabetic mothers (IDM). Methods: Twenty-nine pregnant women with GDM between 37 and 41 gestational weeks who gave birth by spontaneous vaginal delivery were enrolled as study participants together with 20 healthy pregnant women as a control group. Weight, length, and head circumference of babies were measured by the same standard tape immediately after birth. Five milliliters of umbilical venous blood were obtained to study MPV and IMA levels. Results: There was statistically significant difference in levels of MPV (p Z 0.037) and IMA (p < 0.001) between groups. They increased in IDM compared with their healthy peers. Conclusions: Evaluation of MPV and IMA together is useful for representing the potential oxidative stress of IDM. Copyright ª 2014, Taiwan Pediatric Association. Published by Elsevier Taiwan LLC. All rights reserved.
¨ ni* Corresponding author. C ¸ anakkale Onsekiz Mart University Medical Faculty, Department of Pediatrics, C ¸ anakkale Onsekiz Mart U versitesi Tıp Faku ¸ anakkale, Turkey. ¨ltesi Hastanesi, Cumhuriyet Mahallesi Sahil Yolu Number 5, Kepez, C E-mail address: [email protected] (S‚. Yıldırım). http://dx.doi.org/10.1016/j.pedneo.2014.02.002 1875-9572/Copyright ª 2014, Taiwan Pediatric Association. Published by Elsevier Taiwan LLC. All rights reserved. Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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N. Topalo glu et al
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1. Introduction Prevalence of gestational diabetes mellitus (GDM) is increasing worldwide and it is reported to range from 1% to 14% in the literature.1 Race, increasing age, and obesity are the risk factors. Early diagnosis and adequate treatment is important for prevention of complications. While preeclampsia, polyhydramnios, fetal macrosomia, and operative delivery are complications that are seen in GDM pregnant women, hypoglycemia, hypocalcemia, hyperbilirubinemia, and polycytemia are some of the complications seen in the infants of diabetic mothers (IDM). Besides an increment in perinatal mortality, the risk of developing type II diabetes mellitus in the following 10 years increases by 20e30%. Obesity and impaired glucose tolerance ratios increase in the childhood period of IDM.2 Literature about the oxidative stress increasing effect of GDM and decreased antioxidant enzyme capacity in those pregnant women is limited.3 Ischemia modified albumin (IMA) is a biomarker for acute ischemia that is approved by the US Food and Drug Administration. When exposed to ischemic conditions, the N-terminus of albumin is damaged, which makes it unable to bind metals and capable of being measured by an albumin cobalt-binding test. Because its levels in the blood increase within minutes of the onset of ischemia and return to normal within 6e12 hours, IMA has been implicated in the detection of acute ischemia prior to necrosis.4e6 Pregnancy generates a stressful condition in the woman that is related to increased oxidative stress.7 Gugliucci et al demonstrated a marked difference of IMA levels between pregnant and nonpregnant women. IMA levels also increase in normal vaginal delivery.8e10 There is insufficient literature investigating the correlation between GDM and IMA levels. Ma et al found that high levels of IMA and blood glucose levels in GDM pregnant women significantly decreased to normal levels after 6 weeks of regular treatment. 11 The mean platelet volume (MPV) is a useful marker to determine platelet morphology. A normal range of cord blood MPV levels in healthy newborns has been defined.12 Kipper et al12 also revealed the alterations in thrombocyte and IMA level in mature and premature infants. Diabetes also changes thrombocyte function and morphology. Increased MPV is a direct sign of thrombocyte synthesis and activation. MPV increases as a result of thrombocyte synthesis. Sak et al showed a clear increase in the MPV of GDM pregnant women compared with the control group.13 To our knowledge, this is the first study to investigate the cord blood IMA and MPV levels in the same group of diabetic patients. In our study, our aim is to investigate these parameters as a sign of oxidative stress in the IDM.
2. Materials and methods We performed a caseecontrol study from June 2012 to July 2013 at the Departments of Obstetrics and Gynecology and Pediatrics at C ¸ anakkale Onsekiz Mart University Faculty of Medicine. The approval of the ethics committee of our university was obtained. Every patient included in the study signed an informed consent form before participating in the study. We enrolled 29 GDM pregnant women between 37 and 41 gestational weeks who gave birth by spontaneous
vaginal delivery together with 20 healthy pregnant women as a control group. Pregnant women who did not want to enroll in the study, gave birth by cesarean delivery, those who had multiple pregnancies, those with chronic diseases and fetal anomalies, those using chronic medications, and those who smoked were excluded from the study. Postpartum 1st minute and 5th minute APGAR scores were recorded. Babies were weighed without clothes immediately after birth. The length of the baby and head circumference were measured using the same standard tape. Five milliliters of umbilical venous blood were obtained just after delivery into dry tubes with separating gel. Blood was centrifuged at 800 g at 4 C for 15 minutes and the separated serum was immediately analyzed or frozen at 80 C until use. Spectrophotometric measurements were made in a Beckman DU 640 spectrophotometer (Beckman Coulter Inc, Fullerton, CA, USA). IMA was measured by the decrease in cobalt 2þ binding as previously described.4 We introduced minor modifications in the method to adapt it to a 96-well plate reader. Briefly, we added 100 mL of patient serum to 25 mL of a solution of 1 g/L cobalt chloride and a blank for each serum was prepared similarly; þ25 mL of a 9.0 g/L solution of NaCl. After agitation and 10 minutes’ incubation at 25 C, dithiothreitol (25 mL of a 1.5 g/L solution) was then added. After mixing and 2 minutes’ incubation, the absorbance of assay mixtures was read at 470 nm, zeroing with the individual blank. IMA values are expressed in absorbance units (AU). Platelet count and MPV determinations were performed based on the Coulter Counter model LH (Coulter Electronics, Hialeah, FL, USA). Data were recorded using the SPSS 15.0 statistical software package (SPSS Inc., Chicago, IL, USA). Descriptive data are shown as mean standard deviation. ManneWhitney U test was applied for comparisons. A p value < 0.05 was considered significant.
3. Results Table 1 shows the cord blood levels of IMA and MPV in the IDM and control groups. The serum IMA levels were significantly higher in the IDM group compared with the control group (p < 0.001; Figure 1). The mean IMA value was 912.20 92.41 AU in IDM group and 715.60 150.35 AU in the control group. There was also a statistically significant difference between groups in MPV values. The serum MPV was significantly higher in the IDM group (p Z 0.037; Figure 2). In the IDM group, the mean serum MPV was 8.11 0.95 fL and in control group it was 7.55 0.72 fL. Table 1 Cord blood mean platelet volume (MPV) and ischemia modified albumin (IMA) levels of participants. GDM pregnant women
Healthy controls p
MPV, fL 8.11 0.95 7.55 0.72 0.037 Mean SD IMA, absorbance 912.20 92.41 715.60 150.35 <0.001 unit Mean SD Data are presented as the mean standard deviation. GDM Z gestational diabetes mellitus.
Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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MPV and IMA in infants of diabetic mothers
3 Table 2
Characteristics of case and control groups. Case
Control
Gestational week 39.1 0.9 39.0 1.2 Length, cm 49.5 1.6 49.7 1.9 Head circumference, cm 34.5 1.7 34.4 1.34 Birth weight, g 3164 551 3212 509 212.5 68.4 221.9 85.4 Platelet count 1012 Age of mother, y 31.4 5.3 26.8 5.0
p 0.838 0.657 0.967 0.799 0.640 0.004
Data are presented as the mean standard deviation.
Figure 1 Cord blood ischemia-modified albumin (IMA) levels of infants of diabetic mothers and healthy controls. The difference is statistically significant (p < 0.001).
Table 2 represents the demographic characteristics of the newborns and pregnant women in both groups. There was no statistically significant difference between the clinical characteristics of the groups except in maternal age (p > 0.05). The difference in maternal age between groups did not affect the IMA level or MPV.
4. Discussion We found a significant elevation in the cord blood level of IMA and MPV in IDM as evidence of danger to health. Pregnancy generates a hypoxic condition in experimental models. Antioxidative defensive mechanisms change during pregnancy. IMA is used to predict hypoxic conditions, so it may be used in pregnancy to show hypoxemia. Some studies have reported increased IMA levels dependent on oxidative stress triggered by pregnancy.8,9
Figure 2 Cord blood mean platelet volume (MPV) of infants of diabetic mothers and healthy controls. The difference is statistically significant (p Z 0.037).
There is evidence that pregnancy is a physiologically increased stress condition and in complicated pregnancies ¨ zdemir et al IMA levels increase more than normal ones. O found significantly higher levels of IMA in pregnant women with habitual abortions.14 This finding may suggest that an abnormally high hypoxic intrauterine environment may be associated with abnormal placental development that contributes to early miscarriage. Pregnant women carrying fetuses with neural tube defects had higher levels of IMA ¨ stu when compared to normal pregnant women.15 U ¨n et al found that moderate and severe preeclamptic pregnant patients had higher IMA levels than healthy pregnant women and IMA levels positively correlated with the severity of preeclampsia.16 In that study they also showed that healthy and preeclamptic pregnant women had higher IMA levels than nonpregnant women. GDM is a risk for the health of pregnant women and their infants. Women with risk factors should be screened and early diagnosis should be made for prevention of complications. The best test for the diagnosis is oral glucose tolerance test; in our study, all the patients were positive for this test. Oxidative stress had positive correlation with increased blood glucose levels, protein oxidation and lipid peroxidase concentration capacity in GDM pregnant women.17 Ma et al found high levels of IMA in GDM pregnant women compared with healthy ones and they attributed this result to high blood glucose levels.11 In our study, we found higher levels of IMA in cord blood of IDM than the controls. GDM is a systemic disease that affects both mother and fetus.18,19 The platelet count is slightly lower in pregnant than in nonpregnant women.20 Platelet levels also decrease while the duration of pregnancy increases.21 Platelet volume is a marker of platelet activation and function and is measured using the MPV.18,19 MPV values have been found to be higher in diabetic patients when compared to normal controls. When Sharpe and Trinick compared the MPV of GDM and healthy pregnant women, they found significantly higher MPV in GDM pregnant women.22 Kharb23 claimed that low insulin sensitivity is the cause of the oxidative stress and leads to free radical production. Because GDM markedly triggers oxidative stress and MPV directly shows the response of the thrombocytes to the stress, MPV may be used as a marker for oxidative stress. Patients with diabetes have increased platelet activation compared to nondiabetic individuals.19,24,25 MPV values can serve as an effective marker of blood glucose levels.18,26 Recently, Bozkurt et al19 claimed that GDM patients had higher MPV values than normal controls and that patients with high MPV values had low platelet counts. It
Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002
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MODEL
4 has been reported that platelet survival is shorter in diabetic patients.19,26 MPV increment also showed in metabolic syndrome,27 myocardial infarction,28 and stroke29 cases. S‚ahin et al30 found that severe preeclamptic pregnant women had higher MPVs than mild preeclamptic and healthy pregnant women. Large thrombocytes are more active, produce more prothrombotic factors and adhere easily.31 We did not find statistically significant difference between the groups in thrombocyte counts. MPVs are statistically higher in IDM in our study and this finding is supported by the literature. In conclusion, evaluation of MPV and IMA is useful for representing the potential oxidative stress of IDM.
Conflicts of interest Financial disclosure statements have been obtained, and no conflicts of interest have been reported by the authors or by any individuals in control of the content of this article.
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Please cite this article in press as: Topaloglu N, et al., Mean Platelet Volume and Ischemia Modified Albumin Levels in Cord Blood of Infants of Diabetic Mothers, Pediatrics and Neonatology (2014), http://dx.doi.org/10.1016/j.pedneo.2014.02.002