Adult Onset Global Loss of the Fto Gene Alters Body Composition and Metabolism in the Mouse

The strongest BMI–associated GWAS locus in humans is the FTO gene. Rodent studies demonstrate a role for FTO in energy homeostasis and body composition. The phenotypes observed in loss of expression studies are complex with perinatal lethality, stunted growth from weaning, and significant alterations in body composition. Thus understanding how and where Fto regulates food intake, energy expenditure, and body composition is a challenge. To address this we generated a series of mice with distinct temporal and spatial loss of Fto expression. Global germline loss of Fto resulted in high perinatal lethality and a reduction in body length, fat mass, and lean mass. When ratio corrected for lean mass, mice had a significant increase in energy expenditure, but more appropriate multiple linear regression normalisation showed no difference in energy expenditure. Global deletion of Fto after the in utero and perinatal period, at 6 weeks of age, removed the high lethality of germline loss. However, there was a reduction in weight by 9 weeks, primarily as loss of lean mass. Over the subsequent 10 weeks, weight converged, driven by an increase in fat mass. There was a switch to a lower RER with no overall change in food intake or energy expenditure. To test if the phenotype can be explained by loss of Fto in the mediobasal hypothalamus, we sterotactically injected adeno-associated viral vectors encoding Cre recombinase to cause regional deletion. We observed a small reduction in food intake and weight gain with no effect on energy expenditure or body composition. Thus, although hypothalamic Fto can impact feeding, the effect of loss of Fto on body composition is brought about by its actions at sites elsewhere. Our data suggest that Fto may have a critical role in the control of lean mass, independent of its effect on food intake.     

Predictors of Inconsistent Condom Use among a Hard to Reach Population of Young Women with Multiple Sexual Partners in Peri-Urban South Africa

Background

Evidence suggests that multiple concurrent sexual partnering may be a key driver of the high HIV prevalence among young women in South Africa. However, little is known about whether and to what extent women who have multiple sexual partners also engage in other high risk sexual behaviors such as inconsistent condom use. And yet, multiple concurrent sexual partnering is of little epidemiological relevance if all partners in these sexual networks use condoms consistently. This study assesses the prevalence of sexual risk behaviors and HIV, and predictors of inconsistent condom use among women aged 16–24 with multiple sexual partners in a peri-urban setting in South Africa.

Methods

We used Respondent Driven Sampling, a sampling strategy for hard-to-reach populations to recruit 259 women aged 16–24 in a bio-behavioral cross-sectional survey in the Western Cape province. Estimates of population proportions and 95% confidence intervals (CIs) were calculated using the Respondent-Driven Sampling Analysis Tool 5.6 (RDSAT). The primary outcome was inconsistent condom use in the past three months.

Results

Young women reported an average of 7 partners in the past 3 months and a high prevalence of sexual risk behaviors: concurrency (87%), transactional sex (91%) and age mixing (59%). Having >5 sexual partners in the last 3 months doubled the risk of unprotected sex (OR 2.43, CI 1.39–4.25). HIV prevalence was 4% among 16–19 year olds, increasing threefold (12%) at age 20–24.

Discussion

Multiple sexual partnering, where a high number of partners are acquired in a short space of time, is a fertile context for unprotected and risky sexual behavior. The young women featured in this survey present with a constellation of high-risk sexual behaviors that cluster to form a risk syndrome. Carefully tailored repeat bio-behavioral surveillance surveys are recommended for this sub-population.     

Hypothalamic gene expression following ghrelin therapy to gastrectomized rodents

We investigated whether ghrelin depletion (by gastrectomy surgery) and/or treatment/replacement with the gastric hormone ghrelin alters the expression of key hypothalamic genes involved in energy balance, in a manner consistent with ghrelin's pro-obesity effects. At 2 weeks after surgery mice were treated with ghrelin (12 nmol/mouse/day, sc) or vehicle for 8 weeks. Gastrectomy had little effect on the expression of these genes, with the exception of NPY mRNA in the arcuate nucleus that was increased. Ghrelin treatment (to gastrectomized and sham mice) increased the mRNA expression of orexigenic peptides NPY and AgRP while decreasing mRNA expression of the anorexigenic peptide POMC. Two weeks gavage treatment with the ghrelin mimetic, MK-0677, to rats increased NPY and POMC mRNA in the arcuate nucleus and MCH mRNA in the lateral hypothalamus. Thus, while predicted pro-obesity ghrelin signalling pathways were activated by ghrelin and ghrelin mimetics, these were largely unaffected by gastrectomy.     

MARA: a novel approach for highly multiplexed locus-specific SNP genotyping using high-density DNA oligonucleotide arrays

We have developed a locus-specific DNA target preparation method for highly multiplexed single nucleotide polymorphism (SNP) genotyping called MARA (Multiplexed Anchored Runoff Amplification). The approach uses a single primer per SNP in conjunction with restriction enzyme digested, adapter-ligated human genomic DNA. Each primer is composed of common sequence at the 5′ end followed by locus-specific sequence at the 3′ end. Following a primary reaction in which locus-specific products are generated, a secondary universal amplification is carried out using a generic primer pair corresponding to the oligonucleotide and genomic DNA adapter sequences. Allele discrimination is achieved by hybridization to high-density DNA oligonucleotide arrays. Initial multiplex reactions containing either 250 primers or 750 primers across nine DNA samples demonstrated an average sample call rate of ~95% for 250- and 750-plex MARA. We have also evaluated >1000- and 4000-primer plex MARA to genotype SNPs from human chromosome 21. We have identified a subset of SNPs corresponding to a primer conversion rate of ~75%, which show an average call rate over 95% and concordance >99% across seven DNA samples. Thus, MARA may potentially improve the throughput of SNP genotyping when coupled with allele discrimination on high-density arrays by allowing levels of multiplexing during target generation that far exceed the capacity of traditional multiplex PCR.     

Survey of stereotypic behavior in prosimians

Captive animals have been observed to perform a variety of stereotypic behaviors. However, little is known about stereotypic behavior in prosimians. We sent surveys to 96 AZA-accredited institutions to examine stereotypic behavior in these primates. Forty-eight surveys were returned, providing information on 440 individuals of 10 genera. According to the responses, 13.2% of the prosimians surveyed exhibited some form of stereotypic behavior. Pacing was the most common behavior. A logistic regression was used to examine intrinsic characteristics that might influence the performance of stereotypic behavior. The genus of the prosimian was a significant predictor of stereotypic behavior. Individuals of the genus Varecia and Microcebuswere more likely to engage in stereotypic behavior than members of the other genera. Rearing history, age, and sex were not significant predictors of stereotypic behavior. To examine the influence of extrinsic variables on stereotypic behavior, we transformed the data into the percentage of individuals within the enclosure that were reported to exhibit stereotypic behavior, and analyzed them at the enclosure level using a general linear model (GLM) analysis of variance (ANOVA). The only environmental variable that significantly predicted stereotypic behavior was the frequency with which enrichment was provided. Frequent enrichment was provided to those exhibits with a higher percentage of prosimians that engaged in stereotypic behavior. The results of this survey suggest that stereotypic behavior in prosimians may be associated with intrinsic factors (i.e., individual or genus differences) in addition to extrinsic factors related to housing. This knowledge may be helpful in identifying the causes of and effective treatments for stereotypic behavior in prosimians.