mrepo:topics:dnaandbiologicalsamples

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Biological Samples and DNA

Biological samples were collected for the first time in 1999; non-fasting blood samples were used to measure indicators of risk factors for cardiovascular disease (cholesterol, HDL, LDL, triglyceride and glycoslated haemoglobin) and nutritional markers (red cell folate, plasma folate, plasma vitamin B12 and plasma ferritin). The samples also include a blood sample and a cheek swab from which DNA was derived. A further sample was taken and stored for future cell line cultures. Blood was taken using standard phlebotomy techniques using the BD vacutainer system.

Stored biological samples available from 1999 comprise of EDTA plasma, lithium heparin plasma, cell lines, and DNA. The following tables list the current data available from these samples.

Table 1 - Measurements taken in NSHD at 60-64 years
Cardiovascular assessment Vascular structure and function: carotid intima-medial thickness (IMT) and arterial distensibility (GE Vivid-I), carotid/femoral pulse wave velocity (Vicorder) and central blood pressure and pulse wave analysis (Sphygmocor)

Cardiac structure and function: echocardiography (GE Vivid-I). Images from parasternal long axis and short axis views, apical 5-chamber, 4-chamber, 3-chamber, 2-chamber and aortic views (plus conventional and tissue Doppler in 4-chamber view).

Brachial blood pressure (Omron HEM-705), 12 lead ECG (Burdick Eclipse 850i), including 6 minute heart rate and respiration recordings by ECG for heart rate variability measurements.
Musculoskeletal assessment DXA (QDR 4500 Discovery): Hip (total, femoral neck, trochanter, Ward’s), lumbar spine (L1-4), whole body & region BMD, fat & lean mass, vertebral fracture assessment, aortic calcification score

pQCT (Stratec XCT 2000) radius: 4% site(trabecular, cortical and subcortical BMD), 50% site (endosteal/periosteal circumference, cortical CS area & thickness, BMC & BMD, CS muscle & fat area, stress strain index, moment of inertia)
Tests of functional capacity, biological function, and anthropometric measuresVerbal memory, search speed and concentration, simple and choice reaction time, standing balance, grip strength, chair rises, get up and go test, spirometry

Standing and sitting height, weight, chest, upper arm, waist & hip circumference.
Assessment of free-living physical activity Five continuous days by combined heart rate and movement sensor (Actiheart) with individual calibration.
Cardio-respiratory fitness Heart rate response to an incremental step test
Overnight fasting 50ml blood sample See table 2 for details of sample observations
Overnight fasting urine sample Dipstick, spun and unspun aliquots stored at -80°C and -20°C
Salivary samples One salivary sample was collected at the visit and a further three in the following 24 hours (evening, next day waking, and 30 minutes later).
Scales or questions Rose angina, intermittent claudication, bronchitis, chest pain, doctor diagnosed CVD events and test for chest pain/heart disease, osteoarthritis symptoms, back pain, knee injuries, functional limitations, fracture history, prescribed and non-prescribed regular medication, hospital admissions, day surgery, outpatients

Dietary and alcohol assessment (5-day diary), CAGE, smoking, physical activity assessment (EPAQ2), GHQ28, the Close Person Questionnaire, life events, SF36, Edinburgh wellbeing scale, life satisfaction, neighbourhood satisfaction, spare time activities, parental death, time spent caring for others, updated marital and fertility histories

Household income and sources, perceived financial hardship, periods of unemployment, household size and housing tenure, own/partner’s occupation and work status
Table 2 - Biomarkers available at age 53 and 63. Biomarkers highlighted in bold are not present in other UK birth cohorts
System Biomarkers at 53 yrs Biomarkers at 63 yrs
Metabolic Cholesterol, Total (mmol/L) Adiponectin (μg/mL)
Cholesterol, High density Lipoprotein, HDL (mmol/L)Cholesterol, Total (mmol/L)
Cholesterol, Low density Lipoprotein, LDL (mmol/L) Cholesterol, High density Lipoprotein, HDL (mmol/L)
Triglycerides (mmol/L) Cholesterol, Low density Lipoprotein, LDL (mmol/L)
Glycated Haemoglobin, HbA1c (mmol/mol) Triglycerides (mmol/L)
Glucose (nmol/L)
Insulin (pmol/L)
Leptin (ng/mL)
Lipoprotein[a] (mg/dL)
Proinsulin (pmol/L)
Glycated Haemoglobin, HbA1c (mmol/mol)
Diet & Iron Markers Ferritin (ng/mL)
Haemoglobin, Hb (g/L)
Iron (μg/dL)
Vitamin C (μmol/L)
Vitamin D3 (250HD3) (ng/mL)†
Inflammatory Anti-Thyroperoxidase, TPO (IU/mL)
Basophils (x109/L)
C-Reactive Protein, CRP (mg/L)
Eosinophil (x109/L)
Fibrinogen (g/L)
Homocysteine concentration (μmol/L)
Interleukin-6, IL-6 (pg/mL)
Tissue Plasminogen Activator, TPA (ng/mL)
Von Willebrand Factor, VWF (IU/dL)
Red Cell Folate, RCF (nmol/L)
Lymphocytes (x109/L)
Mean Corpuscular Volume, MCV
Mean Corpuscular Haemoglobin, MCH
Mean Platelet Volume, MPV
Monocytes (x109/L)
Neutrophils (x109/L)
Platelet count (x109/L)
Platelet Distribution Width, PDW (%)
Potassium (mmol/L)
Protein (blood), Total (g/L)
Red Blood Cell, RBC, count (109/L)
White Blood Cell, WBC, count (109/L)
Neuro-Endocrine DHEAS (umol/L) Cortisol (saliva) (nmol/L)
Insulin Growth Factor 1, IGF-1 (nmol/L) DHEAS (umol/L)
Sex hormone binding globulin, SHBG (nmol/L) Insulin Growth Factor 1, IGF-1 (nmol/L)
Testosterone (nmol/L) Insulin Growth Factor 2, IGF-2 (nmol/L)
Insulin Growth Factor IGF-BP3 (ng/mL)
Sex hormone binding globulin, SHBG (nmol/L)
Testosterone (nmol/L)
Thyroid tests (T3, T4) (pmol/L)
Thyroid Stimulating Hormone, TSH (mU/L)
Kidney & Liver Calcium (mmol/L)
Creatinine (blood, urine) (mmol/L)
Phosphate (mmol/L)
Urea (nmol/L)
Alanine Transaminase , ALT (IU/L)
Alkaline Phosphatase, ALP (IU/L)
Aspartate Transaminase, AST (IU/L)
Albumin (blood, urine) (mg/L)
Bilirubin (mg/dL)
Cystatin C (mg/L)
Gamma Glutamyl Transferase, GGT (IU/L)
Glucose (urine) (dipstick) (mmol/L)
Globulin (mg/dL)
Ketones (urine) (dipstick) (positive/negative)
Liver Iron Concentration, LIC (%)
Potassium (serum) (mmol/L)
Potassium (urine) (mmol/L)
Protein (urine) (positive/negative)
Sodium (urine) (mmol/L)
Sodium (blood) (mmol/L)
Urate (g/L)

Around 2,900 study members have given consent for DNA to be extracted from blood and/or buccal samples and LHA was given ethical approval in 2007 to use these samples for genetic studies of lifelong health and ageing. Genetic studies help us to elucidate biological mechanisms of ageing and understand why people with similar life experiences age differently.

The NSHD DNA Repository supports high calibre studies of human health and disease that make the best use of the genetic resource in conjunction with the existing life course data. Dr Andrew Wong coordinates the genetic research of the NSHD and is responsible for the safe storage and efficient use of the samples, which are managed by Source BioScience.

Further information on the access to the NSHD DNA Repository is available here.

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