Visceral and subcutaneous adiposity measurements in adults: influence of measurement site

Objective: Excess abdominal adiposity is a known risk factor for cardiovascular diseases. Computed tomography can be used to examine the visceral (VAT) and subcutaneous (SAT) components of abdominal adiposity, but it is unresolved whether single‐slice or multi‐slice protocols are needed. Research Method and Procedures: Nine computed tomography scans were obtained in the lumbar spine region of 24 adults. The nine slices were obtained at three intervertebral positions (L2–L3, L3–L4, and L4–L5) and at 7 mm above and below these locations. Intra‐site and inter‐site differences in SAT, VAT, total adipose tissue, and the VAT/SAT ratio were examined using ANOVA and confidence intervals for pairwise differences between means. Results: Intervertebral SAT values increased from 103.1 ± 50.9 (standard deviation) cm² at L2–L3 to 153.3 ± 68.8 cm² at L4–L5, whereas the corresponding VAT values decreased from 164.3 ± 125.4 to 126.0 ± 82.7 cm². The VAT/SAT ratio was not constant, decreasing from 1.8 ± 1.4 to 0.9 ± 0.7. Repeated‐measures ANOVA indicated significant inter‐ and intra‐site differences (p ≤ 0.02) for SAT, VAT, and the VAT/SAT ratio at L3?L4 and L4?L5 (p < 0.001). Discussion: These differences show the limitation of using a single‐slice assessment of abdominal fat distribution, both for a subject and between subjects. Furthermore, the sizeable differences in the intra‐site scans indicate that precise repositioning is needed for longitudinal studies. In summary, our findings suggest that a multi‐site imaging protocol may provide a more complete assessment of abdominal fat stores and distribution than use of a single site.     

Abdominal fat and risk of coronary heart disease in patients with peripheral arterial disease

Objective: We investigated whether the presence of concomitant coronary heart disease (CHD) in patients with peripheral arterial disease (PAD) can be explained by intra‐abdominal fat accumulation and compared different measures of adiposity as predictors of CHD in patients with PAD. Research Methods and Procedures: Data were collected from patients enrolled in the Second Manifestations of ARTerial disease (SMART) study, an ongoing prospective cohort study of patients with manifest vascular disease or vascular risk factors at the University Medical Centre Utrecht. The current analysis includes 315 patients, mean age 59 ± 10 years, who had PAD with (n = 79) or without (n = 236) CHD. Parameters of adiposity were measured, and intra‐abdominal fat and subcutaneous fat were measured ultrasonographically. Metabolic syndrome was defined according to Adult Treatment Panel III. Results: The prevalence of metabolic syndrome was higher among patients with CHD (63%) than among patients without CHD (48%). All parameters of adiposity indicated more fat in patients with CHD, except for subcutaneous fat. Waist circumference was associated with 64% higher prevalence of CHD (confidence interval, 20% to 123%) per 1 standard deviation increase in waist circumference after adjustment for age and sex. The odds ratio for waist circumference remained virtually the same after additional adjustment for the components of the metabolic syndrome and smoking. Discussion: An increased waist circumference, a crude measure of intra‐abdominal fat, is associated with an increased risk of concomitant CHD in patients with PAD.     

Bottlenecks in domestic animal populations can facilitate the emergence of Trypanosoma cruzi,the aetiological agent of Chagas disease

Faeces-mediated transmission of Trypanosoma cruzi (the aetiological agent of Chagas disease) by triatomine insects is extremely inefficient. Still, the parasite emerges frequently, and has infected millions of people and domestic animals. We synthesize here the results of field and laboratory studies of T. cruzi transmission conducted in and around Arequipa, Peru. We document the repeated occurrence of large colonies of triatomine bugs (more than 1000) with very high infection prevalence (more than 85%). By inoculating guinea pigs, an important reservoir of T. cruzi in Peru, and feeding triatomine bugs on them weekly, we demonstrate that, while most animals quickly control parasitaemia, a subset of animals remains highly infectious to vectors for many months. However, we argue that the presence of these persistently infectious hosts is insufficient to explain the observed prevalence of T. cruzi in vector colonies. We posit that seasonal rains, leading to a fluctuation in the price of guinea pig food (alfalfa), leading to annual guinea pig roasts, leading to a concentration of vectors on a small subpopulation of animals maintained for reproduction, can propel T. cruzi through vector colonies and create a considerable force of infection for a pathogen whose transmission might otherwise fizzle out.     

Comprehensive gene panels provide advantages over clinical exome sequencing for Mendelian diseases

BackgroundTo understand the contribution of Mendelian mutations to the burden of undiagnosed diseases that are suspected to be genetic in origin, we developed a next-generation sequencing-based multiplexing assay that encompasses the ~3000 known Mendelian genes. This assay, which we term the Mendeliome, comprises 13 gene panels based on clinical themes, covering the spectrum of pediatric and adult clinical genetic medicine. We explore how these panels compare with clinical whole exome sequencing (WES).ResultsWe tested 2357 patients referred with suspected genetic diagnoses from virtually every medical specialty. A likely causal mutation was identified in 1018 patients, with an overall clinical sensitivity of 43 %, comparing favorably with WES. Furthermore, the cost of clinical-grade WES is high (typically more than 4500 US dollars), whereas the cost of running a sample on one of our panels is around 75–150 US dollars, depending on the panel. Of the “negative” cases, 11 % were subsequently found by WES to harbor a likely causal mutation in a known disease gene (largely in genes identified after the design of our assay), as inferred from a representative sample of 178. Although our study population is enriched for consanguinity, 245 (24 %) of solved cases were autosomal dominant and 35 (4 %) were X-linked, suggesting that our assay is also applicable to outbred populations.ConclusionsDespite missing a significant number of cases, the current version of the Mendeliome assay can account for a large proportion of suspected genetic disorders, and provides significant practical advantages over clinical WES.

Electronic supplementary material

The online version of this article (doi:10.1186/s13059-015-0693-2) contains supplementary material, which is available to authorized users.     

CEP290 mutations are frequently identified in the oculo-renal form of Joubert syndrome-related disorders

Joubert syndrome–related disorders (JSRDs) are a group of clinically and genetically heterogeneous conditions that share a midbrain-hindbrain malformation, the molar tooth sign (MTS) visible on brain imaging, with variable neurological, ocular, and renal manifestations. Mutations in the CEP290 gene were recently identified in families with the MTS-related neurological features, many of which showed oculo-renal involvement typical of Senior-Löken syndrome (JSRD-SLS phenotype). Here, we performed comprehensive CEP290-mutation analysis on two nonoverlapping cohorts of JSRD-affected patients with a proven MTS. We identified mutations in 19 of 44 patients with JSRD-SLS. The second cohort consisted of 84 patients representing the spectrum of other JSRD subtypes, with mutations identified in only two patients. The data suggest that CEP290 mutations are frequently encountered and are largely specific to the JSRD-SLS subtype. One patient with mutation displayed complete situs inversus, confirming the clinical and genetic overlap between JSRDs and other ciliopathies.     

Association of physical activity and the metabolic syndrome in children and adolescents: CASPIAN Study

BACKGROUND/AIM: To determine the association of physical activity and the metabolic syndrome in a large national-representative sample of children. METHODS: This study was performed in 2003-2004 on 4,811 school students aged 6-18 years, selected by multi-stage random cluster sampling from six provinces in Iran. We assessed the level of physical activity using a standardized questionnaire, and categorized it to the tertiles. The metabolic syndrome was defined based on criteria analogous to those of the Adult Treatment Panel III. RESULTS: The participants comprised 2,248 boys and 2,563 girls with a mean age of 12.07 +/- 3.2 years. In all age groups, boys were more physically active than girls. The metabolic syndrome was detected in 14.1% of participants, and its prevalence was higher in those subjects in the 1st, 2nd and 3rd tertiles of physical activity, respectively (15.1 vs.14.2 and 13.1%, respectively, p <0.05). This difference was seen in boys, while no difference was found between girls in the 2nd and 3rd tertiles of physical activity. Physical activity was linked to a cluster of factors consisting of high-density lipoprotein-cholesterol and waist circumference, followed by triglycerides in boys, and of triglycerides, waist circumference and blood pressure in girls. In both genders, before and after adjustment for age and body mass index, low levels of physical activity significantly increased the risk of having the metabolic syndrome [in boys: OR: 1.8, 95% CI: 1.1, 2.1; and in girls, OR: 1.6 (1.1, 1.9)]. CONCLUSION: We found an association between physical activity and the metabolic syndrome, which was independent of body mass index and age. Children should be encouraged to have greater physical activity.     

Report of the IWGT working group on strategy/interpretation for regulatory in vivo tests II. Identification of in vivo-only positive compounds in the bone marrow micronucleus test

A survey conducted as part of an International Workshop on Genotoxicity Testing (IWGT) has identified a number of compounds that appear to be more readily detected in vivo than in vitro. The reasons for this property varies from compound to compound and includes metabolic differences; the influence of gut flora; higher exposures in vivo compared to in vitro; effects on pharmacology, in particular folate depletion or receptor kinase inhibition. It is possible that at least some of these compounds are detectable in vitro if a specific in vitro test is chosen as part of the test battery, but the 'correct' choice of test may not always be obvious when testing a compound of unknown genotoxicity. It is noted that many of the compounds identified in this study interfere with cell cycle kinetics and this can result in either aneugenicity or chromosome breakage. A decision tree is outlined as a guide for the evaluation of compounds that appear to be genotoxic agents in vivo but not in vitro. The regulatory implications of these findings are discussed.     

World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA

Nucleic acid amplification technique (NAT)-based assays (referred to here as NAT assays) are increasingly used as an alternative to culture-based approaches for the detection of mycoplasma contamination of cell cultures. Assay features, like the limit of detection or quantification, vary widely between different mycoplasma NAT assays. Biological reference materials may be useful for harmonization of mycoplasma NAT assays. An international feasibility study included lyophilized preparations of four distantly related mycoplasma species (Acholeplasma laidlawii, Mycoplasma fermentans, M. orale, M. pneumoniae) at different concentrations which were analyzed by 21 laboratories using 26 NAT assays with a qualitative, semiquantitative, or quantitative design. An M. fermentans preparation was shown to decrease the interassay variation when used as a common reference material. The preparation was remanufactured and characterized in a comparability study, and its potency (in NAT-detectable units) across different NATs was determined. The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) established this preparation to be the “1st World Health Organization international standard for mycoplasma DNA for nucleic acid amplification technique-based assays designed for generic mycoplasma detection” (WHO Tech Rep Ser 987:42, 2014) with a potency of 200,000 IU/ml. This WHO international standard is now available as a reference preparation for characterization of NAT assays, e.g., for determination of analytic sensitivity, for calibration of quantitative assays in a common unitage, and for defining regulatory requirements in the field of mycoplasma testing.