Purification of norovirus-like particles (VLPs) by ion exchange chromatography

Recombinant expression of the norovirus capsid protein VP1 leads to self-assembly of non-infectious virus-like particles (VLPs), which are recognized as promising vaccine candidates against norovirus infections. To overcome the scalability issues connected to the ultracentrifugation-based purification strategies used in previous studies, an anion exchange-based purification method for norovirus VLPs was developed in this study. The method consists of precipitation by polyethylene glycol (PEG) and a single anion exchange chromatography step for purifying baculovirus-expressed GII.4 norovirus VLPs, which can be performed within one day. High product purity was obtained using chromatography. The purified material also contained fully assembled monodispersed VLPs, which were recognized by human sera containing polyclonal antibodies against norovirus GII.4.     

Genetic and environmental determinants of total and high-molecular weight adiponectin in families with low HDL-cholesterol and early onset coronary heart disease

Objective

Plasma adiponectin and high-density lipoprotein cholesterol (HDL-C) exhibit a well-known positive metabolic correlation. Neither heritability nor genome-wide linkage analysis for the high-molecular weight (HMW) adiponectin is available. This work estimates the genetic and environmental determinants and the heritabilities of the adiponectins and lipid traits in Finnish families with early onset coronary heart disease (CHD) and low HDL-C.

Methods

Heritability and genome-wide univariate linkage analysis was performed for total and HMW adiponectin in extended families from Northern Finland with early onset CHD and low HDL-C using a variance components approach. The genetic and environmental correlations between the plasma adiponectins and various lipid traits were also studied and a bivariate analysis for HDL-C and the adiponectins carried out.

Results

In the partial correlation analysis (adjusted for sex, age, BMI and statin use) the adiponectins showed a stronger correlation with HDL-C (total 0.57, p = 0.001, HMW 0.51, p < 0.005) than with any other lipid trait in unrelated subjects. Our estimates detected strong heritability for total (0.53 ± 0.10), HMW (0.51 ± 0.10) and the HMW/total adiponectin ratio (0.68 ± 0.11). Univariate linkage analysis showed suggestive evidence of linkage on chromosome 11p15 for total adiponectin and on 3q13.2-q24 and 6p21 for the HMW adiponectin. The strongest environmental cross-correlation between the adiponectins and lipids was seen between HDL-C and total adiponectin (ρ_e = 0.64, p < 0.05), whereas the strongest genetic correlation was detected between low-density lipoprotein cholesterol and the HMW adiponectin (ρ_g = −0.48, p < 0.05).

Conclusion

No significant genetic correlations between HDL-C and the adiponectins were observed. Therefore, the metabolic association between HDL-C and adiponectin is most likely regulated by complex genetic pathways and environmental factors.     

Longitudinal evaluation of multiple biomarkers for the detection of testosterone gel administration in women with normal menstrual cycle

In women, hormonal fluctuations related to the menstrual cycle may impose a great source of variability for some biomarkers of testosterone (T) administration, which can ultimately disrupt the sensitivity of their longitudinal monitoring. In this study, the sensitivity of the current urinary and haematological markers of the Athlete Biological Passport (ABP), as well as serum steroid biomarkers, was investigated for the monitoring of a 28-day T gel treatment combined with endogenous fluctuation of the menstrual cycle in 14 healthy female subjects. Additionally, the analysis of urinary target compounds was performed on a subset of samples for endogenous/exogenous origin via isotope ratio mass spectrometry (IRMS). In serum, concentrations of T and dihydrotestosterone (DHT) increased significantly during the treatment, whereas in urine matrix the most affected biomarkers were found to be the ratios of testosterone/epitestosterone (T/E) and 5α-androstane-3α,17β-diol/epitestosterone (5αAdiol/E). The detection capability of both urinary biomarkers was heavily influenced by [E], which fluctuated depending on the menstrual cycle, and resulted in low sensitivity of the urinary steroidal ABP module. On the contrary, an alternative approach by the longitudinal monitoring of serum T and DHT concentrations with the newly proposed T/androstenedione ratio showed higher sensitivity. The confirmatory IRMS results demonstrated that less than one third of the tested urine samples fulfilled the criteria for positivity. Results from this study demonstrated that the ‘blood steroid profile’ represents a powerful complementary approach to the ‘urinary module’ and underlines the importance of gathering bundle of evidence to support the scenario of an endogenous prohibited substance administration.     

Annual banned‐substance review: Analytical approaches in human sports drug testing

Several high‐profile revelations concerning anti‐doping rule violations over the past 12 months have outlined the importance of tackling prevailing challenges and reducing the limitations of the current anti‐doping system. At this time, the necessity to enhance, expand, and improve analytical test methods in response to the substances outlined in the World Anti‐Doping Agency's (WADA) Prohibited List represents an increasingly crucial task for modern sports drug‐testing programs. The ability to improve analytical testing methods often relies on the expedient application of novel information regarding superior target analytes for sports drug‐testing assays, drug elimination profiles, alternative test matrices, together with recent advances in instrumental developments. This annual banned‐substance review evaluates literature published between October 2016 and September 2017 offering an in‐depth evaluation of developments in these arenas and their potential application to substances reported in WADA's 2017 Prohibited List.     

Annual banned‐substance review: analytical approaches in human sports drug testing

The annual update of the list of prohibited substances and doping methods as issued by the World Anti‐Doping Agency (WADA) allows the implementation of most recent considerations of performance manipulation and emerging therapeutics into human sports doping control programmes. The annual banned‐substance review for human doping controls critically summarizes recent innovations in analytical approaches that support the efforts of convicting cheating athletes by improved or newly established methods that focus on known as well as newly outlawed substances and doping methods. In the current review, literature published between October 2008 and September 2009 reporting on new and/or enhanced procedures and techniques for doping analysis, as well as aspects relevant to the doping control arena, was considered to complement the 2009 annual banned‐substance review. Copyright © 2010 John Wiley & Sons, Ltd.     

Comparison of management and 30-day mortality of acute myocardial infarction in men versus women in Estonia

OBJECTIVE: There is conflicting information about gender differences in clinical features, management and outcome after acute myocardial infarction (AMI). The objective of the study was to compare the baseline characteristics, management and 30-day mortality of AMI in men and women in Estonia. METHODS: This study included consecutive unselected patients from the Myocardial Infarction Registry (MIR) in Estonia, who were admitted to a university hospital between January 2001 and February 2002. Logistic regression analysis was used to estimate crude and adjusted odds ratios (OR) with 95 percent confidence intervals (95% CI). RESULTS: The study included 228 men and 167 women. Women were older than men (73.49 +/- 10.95 vs. 65.63 +/- 12.60, p < 0.000), and had more comorbidities. After age-adjustment, the higher prevalence of comorbidities, like diabetes (age-adjusted odds ratio [OR] 2.48, 95% confidence intervals [CI] 1.45-4.24), hypertension (OR 1.78, 95% CI 1.15-2.76) and history of congestive heart failure (OR 2.14, 95% CI 1.32-3.46) in women was preserved. Women were more frequently treated with diuretics (OR 2.68, 95% CI 1.69-4.25) and less frequently with statins (OR 0.61, 95% CI 0.39-0.96), after age-adjustment. Although thrombolytic therapy, coronary angiography and angioplasty were performed less frequently in women, these differences disappeared after age-adjustment. Female gender was not an independent predictor of 30-day mortality after AMI, crude OR was 1.39, 95% CI 0.80 to 2.41, adjustment for age and other covariates reduced OR to 0.98, 95% CI 0.44 to 2.20. CONCLUSIONS: Among AMI-patients, age but not gender is an important determinant of care and early mortality.