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Bioassay-guided isolation of cycloartane-type triterpenes, their cytotoxic properties and identification of potential chemical markers from Geniotrigona thoracica propolis

Mohd Badiazaman AA, Nazif Aziz A, Mohamad H, Suryati Mohd K

Nat Prod Res, 2024 May 17.
PMID: 38757417 DOI: 10.1080/14786419.2024.2352867

Phytochemical investigation through cytotoxicity bioassay-guided isolation of Geniotrigona thoracica propolis led to the identification of five cycloartane-type triterpenes, including mangiferonic acid 1, ambonic acid 2, mangiferolic acid 3, ambolic acid 4 and cycloartenol 5. Their structures were established based on detailed spectroscopic analysis and comparison with literature data. Compounds 1-5 were isolated for the first time in this species. From cytotoxicity MTT assay, compound 3 inhibited the growth of breast (MCF-7) and hepatocellular carcinoma (HepG2) cancer cell lines with IC50 of 5.08 and 4.82 µg/mL, respectively. Cytotoxicity of compounds 1-5 were reported for the first time against HepG2 cancer cell lines. Compounds 1-4 were suggested to serve as main and/or analytical markers for G. thoracica propolis based on HPLC analysis. To our knowledge, this is the first report on the isolation of bioactive constituents and identification of chemical markers for G. thoracica propolis.
Fulltext Plasma antioxidants and oxidative stress status in obese women: correlation with cardiopulmonary response

Adenan DM, Jaafar Z, Jayapalan JJ, Abdul Aziz A

PeerJ, 2020;8:e9230.
PMID: 32477840 DOI: 10.7717/peerj.9230

INTRODUCTION: A high body fat coupled with low cardiopulmonary fitness and an increase in oxidative stress has been connoted as contributing factors in developing cardiovascular comorbidities. This study aimed to investigate the correlation between antioxidants and oxidative stress status with cardiopulmonary responses in women of different body mass index (BMI).
SUBJECTS AND METHODS: Eighty female adults were recruited and divided into three groups; normal weight (n = 23), overweight (n = 28) and obese (n = 29), according to their BMI. Blood samples were obtained prior to cardiopulmonary exercise testing. Plasma samples were separated by centrifugation and analysed for enzymatic antioxidant activity including catalase, glutathione peroxidase and superoxide dismutase. Non-enzymatic antioxidant activities were assessed using 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging and ferric reducing ability of plasma (FRAP) assays. To evaluate the oxidative stress status of subjects, levels of reactive oxygen species and malondialdehyde, the by-product of lipid peroxidation, were measured. Cardiopulmonary responses were analysed using cardiopulmonary exercise testing (CPET) which involved 15 various parameters such as peak oxygen consumption, metabolic equivalents and respiratory exchange ratio.
RESULTS: The obese group had significantly lower ABTS radical scavenging and FRAP activities than the normal weight group. A higher catalase activity was observed in the obese group than the normal weight group. Spearman's correlation showed an inverse relationship between catalase and peak oxygen consumption, while partial correlation analysis showed inverse correlations between superoxide dismutase and respiratory frequency, ABTS activity and oxygen pulse, and between ABTS activity and cardiac output.
CONCLUSION: Our results demonstrate a lower cardiovascular fitness and antioxidant capacity in obese women; the higher catalase activity may be a compensatory mechanism. The negative correlations found between these two parameters may indicate the potential effect of antioxidants on the cardiopulmonary system and deserve further analysis in a larger population. Nevertheless, this study provides the basis for future studies to further explore the relationships between redox status and cardiopulmonary responses. This can potentially be used to predict future risk of developing diseases associated with oxidative stress, especially pulmonary and cardiovascular diseases.
Fulltext Attributes Influencing Willingness to Pay for Overweight and Obesity Interventions in Adults: A Systematic Literature Review

Anwar NA, Aizuddin AN, Ahmad N, Aziz A

Cureus, 2024 Jul;16(7):e65468.
PMID: 39071073 DOI: 10.7759/cureus.65468

This review aims to identify the associated attributes of willingness to pay (WTP) for overweight and obesity interventions. A narrative review was conducted by partially adopting the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. A non-exhaustive search using a pre-defined strategy and keywords was done on three selected literature databases: Pubmed, Scopus, and Web of Science. The inclusion criteria for the review were original studies written in English, published between 2000 and 2022, and focused on WTP for overweight and obesity interventions in adults. The extracted studies were manually screened for their eligibility through three cascading tiers: the title, the abstract, and the full article. Only nine original studies were eligible for review based on the screening procedure of 40 screened articles. There was heterogeneity in the study designs, methods, target populations, study duration, and perspectives across the studies. The majority of the studies showed that higher WTP was associated with younger age, having higher income, being female, having higher body mass index (BMI), having the perception of being overweight, habits, and attitudes. WTP is also attributed to the associated percentage of weight loss, long-term health risk reduction, time to noticeable weight loss, delivery mode, side effects, lifestyle modification, and costs of interventions. The identification of common attributes of the WTP for overweight and obesity intervention can assist in the formulation and implementation of effective evidence-based policies. Specific sub-groups with low WTP could be targeted via unique initiatives to improve their participation in weight-loss interventions.
Fulltext Repeated time-series cross-validation: A new method to improved COVID-19 forecast accuracy in Malaysia

Abdul Aziz A, Yusoff M, Yaacob WFW, Mustaffa Z

MethodsX, 2024 Dec;13:103013.
PMID: 39559463 DOI: 10.1016/j.mex.2024.103013

Forecasting COVID-19 cases is challenging, and inaccurate forecast values will lead to poor decision-making by the authorities. Conversely, accurate forecasts aid Malaysian government authorities and agencies (National Security Council, Ministry of Health, Ministry of Finance, Ministry of Education, and Ministry of International Trade and Industry) and financial institutions in formulating action plans, regulations, and legal acts to control COVID-19 spread in the country. Therefore, this study proposes Repeated Time-Series Cross-Validation, a new data-splitting strategy to identify the best forecasting model that is capable of producing the lowest error measures value and a high percentage of forecast accuracy for COVID-19 prediction in Malaysia. Some of the highlights of the proposed method are:•A total of 21 models, five data partitioning sets, and four error measures to improve the forecast accuracy of daily COVID-19 cases in Malaysia.•The best model selected produces the lowest error measure value for the Root Mean Squared Error (RMSE), Mean Absolute Error (MAE), Mean Absolute Percentage Error (MAPE), and Mean Absolute Scaled Error (MASE).•The average 8-day forecast accuracy is 90.2 %. The lowest and highest forecast accuracy was 83.7 % and 98.7 %.
Fulltext Computed tomography of blunt spleen injury: a pictorial review

Hassan R, Abd Aziz A, Md Ralib AR, Saat A

Malays J Med Sci, 2011 Jan;18(1):60-7.
PMID: 22135575 MyJurnal

The spleen is one of the organs most frequently injured in blunt abdominal trauma. Computed tomography (CT) scanning can accurately detect splenic injury and is currently the imaging modality of choice in assessing clinically stable patients with blunt abdominal trauma. The CT features of spleen injury include lacerations, subcapsular or parenchymal haematomas, active haemorrhage, and vascular injuries. We present a pictorial review of the spectrum of CT findings for blunt splenic injuries. This article will be a useful reference for radiologists and surgeons as CT scan is widely used for the assessment of splenic injuries and contributes to the current trend towards nonsurgical management of this injury.
Fulltext Two cases of congenital vallecular cyst: a reminder of the potentially fatal cause of upper airway obstruction in infants

Abd Aziz A, Abdullah AF, Ahmad RA

Malays J Med Sci, 2010 Jul;17(3):68-73.
PMID: 22135553 MyJurnal

Vallecular cyst, a benign yet rare laryngeal lesion, may cause stridor and even life-threatening upper airway obstruction in infants. It can cause apnoea and poor feeding habits, thus reducing the chance of survival. Although laryngomalacia remains the most common cause of stridor in this age group, awareness and a high level of suspicion for this condition can help lead to early management and intervention. Direct laryngoscopy is accepted as the gold standard for diagnostic purposes, and marsupialisation of the cyst is the preferred treatment. We describe 2 cases of vallecular cysts in infants admitted to our hospital where timely diagnoses led to appropriate treatment.
Fulltext Onyx in Brain Arteriovenous Malformation Embolisation

Hashim H, Muda AS, Abdul Aziz A, Abdul Hamid Z

Malays J Med Sci, 2016 Jul;23(4):59-64.
PMID: 27660546 MyJurnal DOI: 10.21315/mjms2016.23.4.8

Embolisation has long been used as an adjunct to surgical resection in the treatment of brain arteriovenous malformation (bAVM). The most commonly used embolic material, n-butylcyanoacrylate glue, requires experience and skill to handle its quick and unpredictable flow and polymerisation. A new liquid embolic agent, ethylene vinyl alcohol copolymer (Onyx), is less adhesive and polymerises slowly, which provides better control for radiologists performing embolisation.
Fulltext The interplay between dysregulated metabolites and signaling pathway alterations involved in osteoarthritis: a systematic review

Aziz A, Ganesan Nathan K, Kamarul T, Mobasheri A, Sharifi A

Ther Adv Musculoskelet Dis, 2024;16:1759720X241299535.
PMID: 39600593 DOI: 10.1177/1759720X241299535

BACKGROUND: Osteoarthritis (OA) is a common degenerative joint disease that poses a significant global healthcare challenge due to its complexity and limited treatment options. Advances in metabolomics have provided insights into OA by identifying dysregulated metabolites and their connection to altered signaling pathways. However, a comprehensive understanding of these biomarkers in OA is still required.
OBJECTIVES: This systematic review aims to identify metabolomics biomarkers associated with dysregulated signaling pathways in OA, using data from various biological samples, including in vitro models, animal studies, and human research.
DESIGN: A systematic review was conducted following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines.
DATA SOURCES AND METHODS: Data were gathered from literature published between August 2017 and May 2024, using databases such as "PubMed," "Scopus," "Web of Science," and "Google Scholar." Studies were selected based on keywords like "metabolomics," "osteoarthritis," "amino acids," "molecular markers," "biomarkers," "diagnostic markers," "inflammatory cytokines," "molecular signaling," and "signal transduction." The review focused on identifying key metabolites and their roles in OA-related pathways. Limitations include the potential exclusion of studies due to keyword selection and strict inclusion criteria.
RESULTS: The meta-analysis identified dysregulated metabolites and associated pathways, highlighting a distinct set of related metabolites consistently altered across the studies analyzed. The dysregulated metabolites, including amino acids, lipids, and carbohydrates, were found to play critical roles in inflammation, oxidative stress, and energy metabolism in OA. Metabolites such as alanine, lysine, and proline were frequently linked to pathways involved in inflammation, cartilage degradation, and apoptosis. Key pathways, including nuclear factor kappa B, mitogen-activated protein kinase, Wnt/β-catenin, and mammalian target of rapamycin, were associated with changes in metabolite levels, particularly in proinflammatory lipids and energy-related compounds.
CONCLUSION: This review reveals a complex interplay between dysregulated metabolites and signaling pathways in OA, offering potential biomarkers and therapeutic targets. Further research is needed to explore the molecular mechanisms driving these changes and their implications for OA treatment.
Fulltext Synthesis, Characterization, and DFT Calculations of a New Sulfamethoxazole Schiff Base and Its Metal Complexes

Al-Hawarin JI, Abu-Yamin AA, Abu-Saleh AAA, Saraireh IAM, Almatarneh MH, Hasan M, et al.

Materials (Basel), 2023 Jul 21;16(14).
PMID: 37512433 DOI: 10.3390/ma16145160

A new Schiff base, 4-((1E,2E)-3-(furan-2-yl)allylidene)amino)-N-(5-methylisoxazol-3-yl) benzene-sulfonamide (L), was synthesized by thermal condensation of 3-(2-furyl)acrolein and sulfamethoxazole (SMX), and the furan Schiff base (L) was converted to a phenol Schiff base (L') according to the Diels-Alder [4 + 2] cycloaddition reaction and studied experimentally. The structural and spectroscopic properties of the Schiff base were also corroborated by utilizing density functional theory (DFT) calculations. Furthermore, a series of lanthanide and transition metal complexes of the Schiff base were synthesized from the nitrate salts of Gd, Sm, Nd, and Zn (L1, L2, L3, and L4), respectively. Various spectroscopic studies confirmed the chemical structures of the Schiff-base ligand and its complexes. Based on the spectral studies, a nine-coordinated geometry was assigned to the lanthanide complexes and a six-coordinated geometry to the zinc complex. The elemental analysis data confirmed the suggested structure of the metal complexes, and the TGA studies confirmed the presence of one coordinated water molecule in the lanthanide complexes and one crystalline water molecule in the zinc complex; in addition, the conductivity showed the neutral nature of the complexes. Therefore, it is suggested that the ligand acts as a bidentate through coordinates to each metal atom by the isoxazole nitrogen and oxygen atoms of the sulfur dioxide moiety of the SMX based on FTIR studies. The ligand and its complexes were tested for their anti-inflammatory, anti-hemolytic, and antioxidant activities by various colorimetric methods. These complexes were found to exhibit potential effects of the selected biological activities.
Fulltext Agricultural intensification, priming for persistence and the emergence of Nipah virus: a lethal bat-borne zoonosis

Pulliam JR, Epstein JH, Dushoff J, Rahman SA, Bunning M, Jamaluddin AA, et al.

J R Soc Interface, 2012 Jan 7;9(66):89-101.
PMID: 21632614 DOI: 10.1098/rsif.2011.0223

Emerging zoonoses threaten global health, yet the processes by which they emerge are complex and poorly understood. Nipah virus (NiV) is an important threat owing to its broad host and geographical range, high case fatality, potential for human-to-human transmission and lack of effective prevention or therapies. Here, we investigate the origin of the first identified outbreak of NiV encephalitis in Malaysia and Singapore. We analyse data on livestock production from the index site (a commercial pig farm in Malaysia) prior to and during the outbreak, on Malaysian agricultural production, and from surveys of NiV's wildlife reservoir (flying foxes). Our analyses suggest that repeated introduction of NiV from wildlife changed infection dynamics in pigs. Initial viral introduction produced an explosive epizootic that drove itself to extinction but primed the population for enzootic persistence upon reintroduction of the virus. The resultant within-farm persistence permitted regional spread and increased the number of human infections. This study refutes an earlier hypothesis that anomalous El Niño Southern Oscillation-related climatic conditions drove emergence and suggests that priming for persistence drove the emergence of a novel zoonotic pathogen. Thus, we provide empirical evidence for a causative mechanism previously proposed as a precursor to widespread infection with H5N1 avian influenza and other emerging pathogens.
Fulltext Polyphenols from the extract and fraction of T. indica seeds protected HepG2 cells against oxidative stress

Razali N, Mat Junit S, Ariffin A, Ramli NS, Abdul Aziz A

BMC Complement Altern Med, 2015;15:438.
PMID: 26683054 DOI: 10.1186/s12906-015-0963-2

Tamarindus indica L. (T. indica) or locally known as "asam jawa" belongs to the family Leguminosae. T. indica seeds as by-products from the fruits were previously reported to contain high polyphenolic content. However, identification of their bioactive polyphenols using recent technologies is less well researched but nonetheless important. Hence, it was the aim of this study to provide further information on the polyphenolic content and antioxidant activities as well as to identify and quantify its bioactive polyphenols.
Fulltext Current trends in water-in-diesel emulsion as a fuel

Yahaya Khan M, Abdul Karim ZA, Hagos FY, Aziz AR, Tan IM

ScientificWorldJournal, 2014;2014:527472.
PMID: 24563631 DOI: 10.1155/2014/527472

Water-in-diesel emulsion (WiDE) is an alternative fuel for CI engines that can be employed with the existing engine setup with no additional engine retrofitting. It has benefits of simultaneous reduction of both NO x and particulate matters in addition to its impact in the combustion efficiency improvement, although this needs further investigation. This review paper addresses the type of emulsion, the microexplosion phenomenon, emulsion stability and physiochemical improvement, and effect of water content on the combustion and emissions of WiDE fuel. The review also covers the recent experimental methodologies used in the investigation of WiDE for both transport and stationary engine applications. In this review, the fuel injection pump and spray nozzle arrangement has been found to be the most critical components as far as the secondary atomization is concerned and further investigation of the effect of these components in the microexplosion of the emulsion is suggested to be center of focus.
Fulltext Effects of Tamarindus indica fruit pulp extract on abundance of HepG2 cell lysate proteins and their possible consequential impact on metabolism and inflammation

Chong UR, Abdul-Rahman PS, Abdul-Aziz A, Hashim OH, Mat-Junit S

Biomed Res Int, 2013;2013:459017.
PMID: 24455694 DOI: 10.1155/2013/459017

The fruit pulp extract of Tamarindus indica has been reported for its antioxidant and hypolipidemic properties. In this study, the methanol extract of T. indica fruit pulp was investigated for its effects on the abundance of HepG2 cell lysate proteins. Cell lysate was extracted from HepG2 cells grown in the absence and presence of the methanol extract of T. indica fruit pulp. Approximately 2500 spots were resolved using two-dimensional gel electrophoresis and the abundance of 20 cellular proteins was found to be significantly reduced. Among the proteins of reduced abundance, fourteen, including six proteins involved in metabolism (including ethanolamine phosphate cytidylyltransferase), four mitochondrial proteins (including prohibitin and respiratory chain proteins), and four proteins involved in translation and splicing, were positively identified by mass spectrometry and database search. The identified HepG2 altered abundance proteins, when taken together and analyzed by Ingenuity Pathways Analysis (IPA) software, are suggestive of the effects of T. indica fruit pulp extract on metabolism and inflammation, which are modulated by LXR/RXR. In conclusion, the methanol fruit pulp extract of T. indica was shown to cause reduced abundance of HepG2 mitochondrial, metabolic, and regulatory proteins involved in oxidative phosphorylation, protein synthesis, and cellular metabolism.
Fulltext Psychometric properties of the 'Skala Kepuasan Interaksi Perubatan-11' to measure patient satisfaction with physician-patient interaction in Malaysia

Abd Aziz A, Izyan Farhana Nordin N, Mohd Noor N, Bachok N, Nor Ismalina Isa S

Fam Pract, 2014 Apr;31(2):236-44.
PMID: 24317538 DOI: 10.1093/fampra/cmt062

Background. Patient satisfaction influences the outcomes of the patient-physician encounter.
Objective. The objective of this study was to validate the Malay version patient satisfaction
(MISS-21) questionnaire using a confirmatory validity approach.
Methods. A cross-sectional study was conducted involving 252 patients attending primary health
clinic, Hospital Universiti Sains Malaysia. Construct validity (convergent and discriminant) using
confirmatory factor analysis and internal consistency were performed after the translation, content
validity and face validity processes. Criterion validity was assessed using Pearson correlations
with the scale of shared decision making 9-item questionnaire (SDMQ-9). The data was
analysed using Analysis of Moment Structure version 19.
Results. A total of 252 (100%) outpatients responded to this study. The final model that consists
of three domains with 11 items had a good fit; (χ2
(df) = 65.805 (32), P
Fulltext Polyphenols in Barringtonia racemosa and their protection against oxidation of LDL, serum and haemoglobin

Kong KW, Mat-Junit S, Ismail A, Aminudin N, Abdul-Aziz A

Food Chem, 2014 Mar 1;146:85-93.
PMID: 24176317 DOI: 10.1016/j.foodchem.2013.09.012

The polyphenolic profiles and antioxidant activities of the water extracts of Barringtonia racemosa shoots (leaves and stems) were explored. Two methods, freeze drying and air drying, for preparation of the shoots, were also compared. Freeze drying was better as air drying caused 5-41% reduction of polyphenols. Three phenolic acids and three flavonoids were identified, using UHPLC. The descending order of polyphenols in the leaves and stems was gallic acid>ellagic acid>quercetin>protocatechuic acid>rutin>kaempferol. In vitro antioxidant analyses were performed using biological samples. In the LDL oxidation assay, B. racemosa leaf extract (IC50=73.0μg/ml) was better than stem extract (IC50=226μg/ml) at inhibiting the formation of TBARS and lipid hydroperoxides. Similar trends were observed for serum and haemoglobin oxidation. B. racemosa leaf extract was better than its stem extract in delaying the time required to oxidise haemoglobin to methaemoglobin. The high polyphenolic content of B. racemosa shoots could have contributed towards their antioxidative effects.
Fulltext Knowledge of Good Blood Culture Sampling Practice among Healthcare Staffs in An Emergency Department - Are We Getting It Right?

Chew KS, Mohd Hashairi F, Jusoh AF, Aziz AA, Nik Hisamuddin NAR, Siti Asma H

Med J Malaysia, 2013 Aug;68(4):323-5.
PMID: 24145260 MyJurnal

Although a vital test, blood culture is often plagued with the problem of contamination and false results, especially in a chaotic emergency department setting. The objectives of this pilot study is to find out the level of understanding among healthcare staffs in emergency department, Hospital Universiti Sains Malaysia (HUSM) regarding good blood culture sampling practice.
Fulltext Tamarindus indica extract alters release of alpha enolase, apolipoprotein A-I, transthyretin and Rab GDP dissociation inhibitor beta from HepG2 cells

Chong UR, Abdul-Rahman PS, Abdul-Aziz A, Hashim OH, Junit SM

PLoS One, 2012;7(6):e39476.
PMID: 22724021 DOI: 10.1371/journal.pone.0039476

The plasma cholesterol and triacylglycerol lowering effects of Tamarindus indica extract have been previously described. We have also shown that the methanol extract of T. indica fruit pulp altered the expression of lipid-associated genes including ABCG5 and APOAI in HepG2 cells. In the present study, effects of the same extract on the release of proteins from the cells were investigated using the proteomics approach.
Differential regulation of fatty acid biosynthesis in two Chlorella species in response to nitrate treatments and the potential of binary blending microalgae oils for biodiesel application

Cha TS, Chen JW, Goh EG, Aziz A, Loh SH

Bioresour Technol, 2011 Nov;102(22):10633-40.
PMID: 21967717 DOI: 10.1016/j.biortech.2011.09.042

This study was undertaken to investigate the effects of different nitrate concentrations in culture medium on oil content and fatty acid composition of Chlorella vulgaris (UMT-M1) and Chlorella sorokiniana (KS-MB2). Results showed that both species produced significant higher (p<0.05) oil content at nitrate ranging from 0.18 to 0.66 mM with C. vulgaris produced 10.20-11.34% dw, while C. sorokiniana produced 15.44-17.32% dw. The major fatty acids detected include C16:0, C18:0, C18:1, C18:2 and C18:3. It is interesting to note that both species displayed differentially regulated fatty acid accumulation patterns in response to nitrate treatments at early stationary growth phase. Their potential use for biodiesel application could be enhanced by exploring the concept of binary blending of the two microalgae oils using developed mathematical equations to calculate the oil mass blending ratio and simultaneously estimated the weight percentage (wt.%) of desirable fatty acid compositions.
Fulltext A mimicry of melioidosis by Klebsiella ozaenae infection

Ng TH, How SH, Kuan YC, Adzura, Aziz AA, Fauzi AR

Malays J Pathol, 2009 Dec;31(2):147-50.
PMID: 20514860 MyJurnal

Klebsiella ozaenae is a Gram negative bacillus. It has been described as a colonizer of oral and nasopharyngeal mucosa and is a cause of atrophic rhinitis. Klebsiella ozaenae has seldom been isolated from serious infections. However, several reports have stated that Klebsiella ozaenae may cause invasive infections and even mortality. We report a 55-year-old man with Klebsiella ozaenae infection causing abscesses involving the right eye and left kidney and possibly also in the brain, lungs and prostate. The isolates were sensitive to ceftazidime, ciprofloxacin, chloramphenicol, gentamicin and sulfamethoxazole-trimethoprim but resistant to ampicillin. He responded well to 4 weeks of i.v. ceftazidime and i.v. amoxycillin-clavulanic acid. To our knowledge, such a multiorgan infection has not been reported previously for this organism.
Fulltext An alternate method for DNA and RNA extraction from clotted blood

Zakaria Z, Umi SH, Mokhtar SS, Mokhtar U, Zaiharina MZ, Aziz AT, et al.

Genet. Mol. Res., 2013;12(1):302-11.
PMID: 23408417 DOI: 10.4238/2013.February.4.4

We developed an alternative method to extract DNA and RNA from clotted blood for genomic and molecular investigations. A combination of the TRIzol method and the QIAamp spin column were used to extract RNA from frozen clotted blood. Clotted blood was sonicated and then the QIAamp DNA Blood Mini Kit was used for DNA extraction. Extracted DNA and RNA were adequate for gene expression analysis and copy number variation (CNV) genotyping, respectively. The purity of the extracted RNA and DNA was in the range of 1.8-2.0, determined by absorbance ratios of A(260):A(280). Good DNA and RNA integrity were confirmed using gel electrophoresis and automated electrophoresis. The extracted DNA was suitable for qPCR and microarrays for CNV genotyping, while the extracted RNA was adequate for gene analysis using RT-qPCR.

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