Myths and Methodologies: Standardisation in Human Physiology Research-Should We Control the Controllables?

The premise of research in human physiology is to explore a multifaceted system whilst identifying one or a few outcomes of interest. Therefore, the control of potentially confounding variables requires careful thought regarding the extent of control and complexity of standardisation. One common factor to control prior to testing is diet, as food and fluid provision may deviate from participants' habitual diets, yet a self-report and replication method can be flawed by under-reporting. Researchers may also need to consider standardisation of physical activity, whether it be through familiarisation trials, wash-out periods, or guidance on levels of physical activity to be achieved before trials. In terms of pharmacological agents, the ethical implications of standardisation require researchers to carefully consider how medications, caffeine consumption and oral contraceptive prescriptions may affect the study. For research in females, it should be considered whether standardisation between- or within-participants in regards to menstrual cycle phase is most relevant. The timing of measurements relative to various other daily events is relevant to all physiological research and so it can be important to standardise when measurements are made. This review summarises the areas of standardisation which we hope will be considered useful to anyone involved in human physiology research, including when and how one can apply standardisation to various contexts.

Myths and Methodologies: Standardisation in human physiology research-should we control the controllables?

The premise of research in human physiology is to explore a multifaceted system whilst identifying one or a few outcomes of interest. Therefore, the control of potentially confounding variables requires careful thought regarding the extent of control and complexity of standardisation. One common factor to control prior to testing is diet, as food and fluid provision may deviate from participants' habitual diets, yet a self-report and replication method can be flawed by under-reporting. Researchers may also need to consider standardisation of physical activity, whether it be through familiarisation trials, wash-out periods, or guidance on levels of physical activity to be achieved before trials. In terms of pharmacological agents, the ethical implications of standardisation require researchers to carefully consider how medications, caffeine consumption and oral contraceptive prescriptions may affect the study. For research in females, it should be considered whether standardisation between- or within-participants in regards to menstrual cycle phase is most relevant. The timing of measurements relative to various other daily events is relevant to all physiological research and so it can be important to standardise when measurements are made. This review summarises the areas of standardisation which we hope will be considered useful to anyone involved in human physiology research, including when and how one can apply standardisation to various contexts.

Are there sex differences in the variability of fasting metabolism?

There is evidence across species and across many traits that males display greater between-individual variance. In contrast, (premenopausal) females display large within-individual variance in sex hormone concentrations, which can increase within-individual variance in many other parameters. The latter may contribute to the lower representation of females in metabolic research. This study is a pooled secondary analysis of data from seven crossover studies to investigate the between-individual and the within-individual variance in fasting plasma metabolites, resting metabolic rate (RMR), and body mass. Females demonstrated higher within-individual variability of plasma 17ß-estradiol [coefficient of variation (CV): 15 ± 15% for males vs. 38 ± 34% for females, P < 0.001] and progesterone concentrations (CV: 13 ± 11% for males vs. 52 ± 51% for females, P < 0.001) but there were no meaningful differences in the variability of plasma glucose (CV: 4 ± 3% for males vs. 5 ± 5% for females), insulin, lactate, triglycerides (CV: 15 ± 9% for males vs. 15 ± 10% for females), and esterified fatty acid concentrations or in RMR and body mass (CV: 0.43 ± 0.34% for males vs. for 0.42 ± 0.33% females; P > 0.05 for all outcomes). Males displayed higher between-individual variance in RMR compared with females (SD: 224 kcal·day-1 for males vs. 151 kcal·day-1 for females). In conclusion, these data do not provide evidence that females show greater within-individual variability in many fasting metabolic variables, RMR, or body mass compared with males. We conclude that including females in metabolic research is unlikely to introduce greater within-individual variance when using the recruitment and control procedures described in these studies.NEW & NOTEWORTHY To investigate the within-individual variability in metabolic parameters in males and females, we performed a pooled secondary analysis of fasting blood samples, resting metabolic rate, and body mass from seven crossover studies. We found a greater day-to-day variation in 17ß-estradiol and progesterone in females compared with males but no meaningful difference in within-individual variability of fasting plasma glucose, insulin, lactate, triglycerides, NEFA, resting metabolic rate, or body mass between females and males.

The effect of exercise in a fasted state on plasma low-density lipoprotein cholesterol concentrations in males and females.

Cardiovascular disease (CVD) is the leading cause of death worldwide. Physical activity interventions improve almost all modifiable CVD risk factors, but the effect of physical activity on low density lipoprotein cholesterol (LDL-C) is uncertain. This may be due to lack of research on the feeding status in which the physical activity is performed. The aim of this study is to investigate the effect of fasted versus fed exercise on LDL-C concentrations in males and females. One hundred healthy participants, equal males and females, aged between 25 and 60 years will be recruited and will undergo a home-based 12-week exercise intervention. After baseline testing, participants will be randomized to a fasted exercise (exercise after an 8-h fast) or fed exercise (exercise 90-180 min after ingestion of 1 g kg-1 CHO) group and will perform 50 min of moderate intensity exercise (e.g., 95% heart rate of lactate threshold 1) three times a week either before or after a high carbohydrate (1 g kg-1 ) meal. Participants will visit the laboratory again at week 4 and week 12 and measurements will be taken for body composition, resting blood pressure, fasting blood glucose, lipid profiles and systemic inflammation, lactate threshold, and 14-day blood glucose control.

Changes in human health parameters associated with a touch tank experience at a zoological institution.

Association of Zoos and Aquariums (AZA) institutions provide a variety of benefits to visitors. However, one area that has received little study is the direct human health benefits from zoo and aquarium visits. With the increase in stress related non-infectious diseases in industrialized countries, understanding the extent of these benefits is important. We studied the effects on visitor stress of an experience at a touch tank exhibit featuring stingrays, sharks, and horseshoe crabs. Stress was measured by physiological and psychological parameters. Heart rate was recorded before, during, and after interacting with the animals, and mood was assessed before and after the experience using a psychological instrument. Multilevel models of heart rate show a quadratic trend, with heart rate elevated (b = -3.01, t = 26.4, P < 0.001) and less variable (b = 3.60, t = 15.9, P < 0.001) while touching the animals compared to before or after. Wilcoxon signed-rank tests on mood data suggest that most visitors felt happier (V = 174.5, P < 0.001), more energized (V = 743.5, P < 0.001), and less tense (V = 5618, P < 0.001) after the experience. This suggests that interacting with animals led to a physiological response during interactions reminiscent of a theme park experience along with a decrease in mental stress. The effects of confounding variables such as crowd size are also discussed. Further studies should be conducted to help deepen our understanding of the health benefits of experiences at AZA institutions.

The role of the interferon gamma release assay in assessing recent tuberculosis transmission in a hospital incident.

In 2007, an extensive contact screening investigation into onward transmission of tuberculosis was instigated at a hospital in Northern Ireland following diagnosis of pulmonary multi-drug resistant TB in a healthcare worker. Interferon gamma release assays (IGRAs) were used to test 333 patients and 98 staff. We investigated for evidence of onward transmission and recent infection based on analysis of clinical, demographic and IGRA data. We also described within-patient variability of IGRA results. Among patients and staff, increasing age of patients was the only factor associated with IGRA positivity. Greatest within-subject variability of IU/mL in serially-tested patients/staff was seen in those with a positive IGRA test and this did not correlate with increased exposure to the index case. IGRA positivity being largely explained by increasing age in patients and previous TB contact in staff lends weight to the conclusion that IGRA positivity reflected previous infection rather than recent transmission.

Investigating transmission of Mycobacterium bovis in the United Kingdom in 2005 to 2008.

Due to an increase in bovine tuberculosis in cattle in the United Kingdom, we investigated the characteristics of Mycobacterium bovis infection in humans and assessed whether extensive transmission of M. bovis between humans has occurred. A cross-sectional study linking demographic, clinical, and DNA fingerprinting (using 15-locus mycobacterial interspersed repetitive-unit-variable-number tandem-repeat [MIRU-VNTR] typing) data on cases reported between 2005 and 2008 was undertaken. A total of 129 cases of M. bovis infection in humans were reported over the period, with a decrease in annual incidence from 0.065 to 0.047 cases per 100,000 persons. Most patients were born pre-1960, before widespread pasteurization was introduced (73%), were of white ethnicity (83%), and were born in the United Kingdom (76%). A total of 102 patients (79%) had MIRU-VNTR typing data. A total of 31 of 69 complete MIRU-VNTR profiles formed eight distinct clusters. The overall clustering proportion determined using the n - 1 method was 33%. The largest cluster, comprising 12 cases, was indistinguishable from a previously reported West Midlands outbreak strain cluster and included those cases. This cluster was heterogeneous, having characteristics supporting recent zoonotic and human-to-human transmission as well as reactivation of latent disease. Seven other, smaller clusters identified had demographics supporting recrudescence rather than recent infection. A total of 33 patients had incomplete MIRU-VNTR profiles, of which 11 may have yielded 2 to 6 further small clusters if typed to completion. The incidence of M. bovis in humans in the United Kingdom remains low, and the epidemiology is predominantly that of reactivated disease.