A model system in landscape genetics – the insect inhabitants of pitcher plants

Landscape genetics is a rapidly growing field that investigates how landscape and environmental features interact with microevolutionary processes to give rise to spatial genetic variation in populations. The ability to predict landscape effects on genetic patterns has been limited by the lack of studies conducted on more than one species, over multiple spatial scales and in replicated landscapes. A system that allows for such studies are the insect inhabitants of the purple pitcher plant (Sarracenia purpurea): the pitcher plant flesh fly (Fletcherimyia fletcheri), the midge (Metriocnemus knabi) and the mosquito (Wyeomyia smithii). In this thesis, I worked towards developing this as a potential model system in landscape genetics. I successfully developed microsatellite markers for the flesh fly and the midge. In the pitcher plant mosquito microsatellite isolation was very problematic, due to presence of microsatellite families associated with transposable elements and further aggravated by null alleles. I found that the pitcher plant flesh fly exhibits a mixture of metapopulation and patchy population attributes. I found that the pitcher plant midge exhibits a complex pattern of genetic differentiation across spatial scales, significantly associated with landscape variables related to habitat size, abundance and spatial arrangement. Also, in this small insect, both active flight and wind-assisted dispersal mediate gene flow among bogs within a landscape. Overall, this research has laid a foundation for further studies in this system and provided insights that are of interest to the broader community of landscape genetics researchers

Impaired Insulin sensitivity and Insulin secretion in Haemodialysis patients with and without Secondary Hyperparathyroidism

The aim of our study was to investigate insulin sensitivity and beta cell function in hemodialysis (HD) patients without diabetes. We hypothesized that parathyroid gland function was a determinant of insulin sensitivity and/or beta cell function. The study was a randomized, cross-sectional one and patients were divided into two groups (total 27 patients), Gp.1 being those with relative hypoparathyroidism (iPTH<200 pg/ml) ­ 9 (33.3%), Gp.2 those with hyperparathyroidism (iPTH200 pg/ml) ­ 18 (66.6%) with Gp.3 (consisting of 43 healthy subjects acting as controls). Insulin resistance and insulin secretion were calculated from fasting serum insulin and glucose concentrations by the Homeostatic Model Assessment score (HOMA IR and HOMA BETA). The value of HOMA IR (3.28±1.3 for Gp.1, 4.80±2.4 for Gp.2, 1.70±0.8 for Gp.3) as well as the glucose level (5.0±1.0mmol/l in Gp.1, 5.2±0.8mmol/ l in Gp.2, 4.6±0.4mmol/l in Gp.3) was significantly higher in HD patients than in control subjects. Excessive insulin secretion was present in HD patients (as assessed by HOMA BETA) significantly higher only in Gp.1 (p=0.02).peer-reviewe

A rapid and reliable determination of doxycycline hyclate by HPLC with UV detection in pharmaceutical samples

An accurate, sensitive and reproducible high performance liquid chromatographic (HPLC) method for the quantification of doxycycline hyclate in pharmaceutical samples has been developed and validated. The drug and the standard were eluted from a Lichrosorb RP-8 (250 mm´4.6 mm, 10 mm particle size) at 20 °C with a mobile phase consisting of methanol, acetonitrile and 0.010 M aqueous solution of oxalic acid (2:3:5, v/v/v). The flow rate was 1.25 ml min-1. A UV detector set at 350 nm was used to monitor the effluent. Each analysis required no longer than 4 min. The limits of detection and quantification were 1.15 and 3.84 μg ml-1, respectively. Recoveries for different concentrations ranged from 99.58 to 101.93 %

Local introduction and heterogeneous spatial spread of dengue-suppressing Wolbachia through an urban population of Aedes Aegypti

Dengue-suppressing Wolbachia strains are promising tools for arbovirus control, particularly as they have the potential to self-spread following local introductions. To test this, we followed the frequency of the transinfected Wolbachia strain wMel through Ae. aegypti in Cairns, Australia, following releases at 3 nonisolated locations within the city in early 2013. Spatial spread was analysed graphically using interpolation and by fitting a statistical model describing the position and width of the wave. For the larger 2 of the 3 releases (covering 0.97 km2 and 0.52 km2), we observed slow but steady spatial spread, at about 100–200 m per year, roughly consistent with theoretical predictions. In contrast, the smallest release (0.11 km2) produced erratic temporal and spatial dynamics, with little evidence of spread after 2 years. This is consistent with the prediction concerning fitness-decreasing Wolbachia transinfections that a minimum release area is needed to achieve stable local establishment and spread in continuous habitats. Our graphical and likelihood analyses produced broadly consistent estimates of wave speed and wave width. Spread at all sites was spatially heterogeneous, suggesting that environmental heterogeneity will affect large-scale Wolbachia transformations of urban mosquito populations. The persistence and spread of Wolbachia in release areas meeting minimum area requirements indicates the promise of successful large-scale population transf

Releasing incompatible males drives strong suppression across populations of wild and Wolbachia-carrying Aedes aegypti in Australia

Releasing sterile or incompatible male insects is a proven method of population management in agricultural systems with the potential to revolutionize mosquito control. Through a collaborative venture with the “Debug” Verily Life Sciences team, we assessed the incompatible insect technique (IIT) with the mosquito vector Aedes aegypti in northern Australia in a replicated treatment control field trial. Backcrossing a US strain of Ae. aegypti carrying Wolbachia wAlbB from Aedes albopictus with a local strain, we generated a wAlbB2-F4 strain incompatible with both the wild-type (no Wolbachia) and wMel-Wolbachia Ae. aegypti now extant in North Queensland. The wAlbB2-F4 strain was manually mass reared with males separated from females using Verily sex-sorting technologies to obtain no detectable female contamination in the field. With community consent, we delivered a total of three million IIT males into three isolated landscapes of over 200 houses each, releasing ∼50 males per house three times a week over 20 wk. Detecting initial overflooding ratios of between 5:1 and 10:1, strong population declines well beyond 80% were detected across all treatment landscapes when compared to controls. Monitoring through the following season to observe the ongoing effect saw one treatment landscape devoid of adult Ae. aegypti early in the season. A second landscape showed reduced adults, and the third recovered fully. These encouraging results in suppressing both wild-type and wMel-Ae. aegypti confirms the utility of bidirectional incompatibility in the field setting, show the IIT to be robust, and indicate that the removal of this arbovirus vector from human-occupied landscapes may be achievable

Data from: The pitcher plant flesh fly exhibits a mixture of patchy and metapopulation attributes

We investigated the pattern of spatial genetic structure and the extent of gene flow in the pitcher plant flesh fly Fletcherimyia fletcheri, the largest member of the inquiline community of the purple pitcher plant Sarracenia purpurea. Using microsatellite loci, we tested the theoretical predictions of different hypothesized population models (patchy population, metapopulation or isolated populations) among 11 bogs in Algonquin Provincial Park (Canada). Our results revealed that the pitcher plant flesh fly exhibits a mixture of patchy and metapopulation characteristics. There is significant differentiation among bogs and limited gene flow at larger spatial scales, but local populations do not experience frequent local extinctions/recolonizations. Our findings suggest a strong dispersal ability and stable population sizes in F. fletcheri, providing novel insights into the ecology of this member of a unique ecological microcosm

F. fletcheri microsatellite genotyping data

Individuals (e.g. ff2009-DL-10A) genotyped at 10 microsatellite loci (e.g. FF72), with one allele per column for each locus. The year of collection is designated after ff (e.g. ff2009 for the collection year 2009), followed by the bog code (e.g. DL), followed by an unique plant number (e.g. 10), followed by a leaf number where multiple individuals from the same plant are collected (e.g. A). The list of bog codes is found in Table 2 of the main text

Rasic&Keyghobadi_Supl_Tables

Supplemental Tables 1-

Omega-3 Polyunsaturated Fatty Acids Enriched Hen Eggs Consumption Enhances Microvascular Reactivity in Young Healthy Individuals

Whilst the beneficial effect of omega-3 polyunsaturated fatty acids (PUFAs) supplementation on cardiovascular (CV) system is well supported in CV patients, the effect of consumption of omega-3 PUFAs enriched functional food in healthy individuals is still not fully elucidated. This study aimed to determine the effect of consumption of omega-3 PUFAs enriched hen eggs on microvascular reactivity (primary outcome), blood pressure (BP) and serum lipid profile in young healthy individuals. Control group (N=16) ate three ordinary hen eggs (277 mg omega-3 PUFAs/day), and OMEGA-3 group (N=20) ate three omega-3 PUFAs enriched eggs containing 259 mg of omega-3 PUFAs/egg daily (ALA 167 mg/egg, EPA 7 mg/egg, DHA 84 mg/egg) for 3 weeks (777 mg omega-3 PUFAs/day). Post-occlusive reactive hyperemia (PORH) in skin microcirculation assessed by laser Doppler flowmetry, serum lipid profile, fasting blood glucose, high-sensitivity C-reactive protein (hsCRP) and arterial BP were measured in all subjects before and after the protocol. PORH was significantly enhanced, and triglycerides, hsCRP and BP were significantly decreased in OMEGA-3 group compared to baseline measurement, while there was no significant difference in Control group after the protocol compared to baseline. This is the first study to demonstrate that consumption of a mixture of omega-3 PUFAs (ALA+EPA+DHA), provided via enriched hen eggs, elicits changes in microvascular reactivity, BP and triglycerides level in healthy subjects that are associated with CV benefits, thus suggesting that daily consumption of omega-3 PUFAs enriched eggs in healthy individuals may potentially contribute to CV risk factors attenuation and disease prevention.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author