Effect of Metabolic Risk Factors, Gene Polymorphisms …
145
2 Materials and Methods
In this study, 100 healthy adult Asian Indian women population, including 50 without
FHD and 50 with FHD, were used to determine the risk for T2DM for both mothers
and their future generation. For this purpose, both mothers’ venous blood and cord
blood of their babies were used to determine their risk associated with T2DM by
taking biochemical measurements such as fasting blood sugar (7 ml), CRP for both
mother and their babies, triglycerides, HbA1C, and HDL cholesterol by auto-analyzer
(Robonik India) as per NECP ATP Panel-III (International Diabetes Federation 2006)
guidelines. This study was approved by the “Institutional Ethics Committee of the
Heritage Institute of Technology”.
Genomic DNA for all samples was isolated using DNA purification spin kit and
then amplified by using PCR (ABI Biosystems USA). For IRS-1, 221 bp PCR product
containing the polymorphic site (Gly972Arg) was digested with BstNI (Ijaz et al.
2019) and obtained the following bands: wild type homozygotes (GG)—two bands
at 31 and 190 bp, variant homozygotes (AA)—one band at 190 bp, and heterozygous
(GA)—three bands at 31, 190, and 221 bp. For IRS-2, 291 bp PCR product containing
the polymorphic site (Gly1057Asp) was digested with HhaI (Bodhini et al. 2007;
Ijaz et al. 2019) and obtained the bands showed at 268 bp and 23 bp, respectively.
For CRP, a 744 bp PCR product containing the polymorphic site (1059 G > C) was
digested by MaeIII (Thalmaier et al. 2006) and obtained the following band: · digestion
of the less common 1059 C allele produced two smaller fragments with sizes of 434
and 310 bp. Digestion of the more common 1059G allele produced three fragments
with sizes of 310, 233, and 201 bp, respectively. Amplified products were separated
by electrophoresis on a 1.5% agarose gel containing ethidium bromide and bands
were then visualized by UV transilluminator (Kodak Ultra-cam Imaging, Japan).
3 Results and Discussion
3.1 Biochemical Analysis of Metabolic Risk Factors
Statistical analysis of the comparison of group differences (with and without FHD)
for all the variables (12 and 30 weeks of gestational period) was performed using
t-test (p < 0.05). Table 1 shows the biochemical parameters which were considered
during the study.
A comparison of the biochemical characteristics among the subjects with and
without FHD by two-tailed t-test revealed that there were significant differences
among them with respect to triglycerides (TG), total cholesterol (TC), fasting blood
sugar (FBS), HDL cholesterol, HbA1c (glycated haemoglobin), and C-reactive
protein (CRP), both at the gestational period of 12 and 30 weeks, respectively. This
indicates that family history plays a pivotal role in influencing metabolic variables
145
2 Materials and Methods
In this study, 100 healthy adult Asian Indian women population, including 50 without
FHD and 50 with FHD, were used to determine the risk for T2DM for both mothers
and their future generation. For this purpose, both mothers’ venous blood and cord
blood of their babies were used to determine their risk associated with T2DM by
taking biochemical measurements such as fasting blood sugar (7 ml), CRP for both
mother and their babies, triglycerides, HbA1C, and HDL cholesterol by auto-analyzer
(Robonik India) as per NECP ATP Panel-III (International Diabetes Federation 2006)
guidelines. This study was approved by the “Institutional Ethics Committee of the
Heritage Institute of Technology”.
Genomic DNA for all samples was isolated using DNA purification spin kit and
then amplified by using PCR (ABI Biosystems USA). For IRS-1, 221 bp PCR product
containing the polymorphic site (Gly972Arg) was digested with BstNI (Ijaz et al.
2019) and obtained the following bands: wild type homozygotes (GG)—two bands
at 31 and 190 bp, variant homozygotes (AA)—one band at 190 bp, and heterozygous
(GA)—three bands at 31, 190, and 221 bp. For IRS-2, 291 bp PCR product containing
the polymorphic site (Gly1057Asp) was digested with HhaI (Bodhini et al. 2007;
Ijaz et al. 2019) and obtained the bands showed at 268 bp and 23 bp, respectively.
For CRP, a 744 bp PCR product containing the polymorphic site (1059 G > C) was
digested by MaeIII (Thalmaier et al. 2006) and obtained the following band: · digestion
of the less common 1059 C allele produced two smaller fragments with sizes of 434
and 310 bp. Digestion of the more common 1059G allele produced three fragments
with sizes of 310, 233, and 201 bp, respectively. Amplified products were separated
by electrophoresis on a 1.5% agarose gel containing ethidium bromide and bands
were then visualized by UV transilluminator (Kodak Ultra-cam Imaging, Japan).
3 Results and Discussion
3.1 Biochemical Analysis of Metabolic Risk Factors
Statistical analysis of the comparison of group differences (with and without FHD)
for all the variables (12 and 30 weeks of gestational period) was performed using
t-test (p < 0.05). Table 1 shows the biochemical parameters which were considered
during the study.
A comparison of the biochemical characteristics among the subjects with and
without FHD by two-tailed t-test revealed that there were significant differences
among them with respect to triglycerides (TG), total cholesterol (TC), fasting blood
sugar (FBS), HDL cholesterol, HbA1c (glycated haemoglobin), and C-reactive
protein (CRP), both at the gestational period of 12 and 30 weeks, respectively. This
indicates that family history plays a pivotal role in influencing metabolic variables