An acid metabolite of cyclosporine

The primary biliary metabolite of cyclosporine has been isolated from rabbit and human bile. The material has been identified by mass spectrometry and by nuclear magnetic resonance spectrometry as an acidic metabolite of cyclosporine in which the eta-methyl group of the cyclosporine-specific nine carbon amino acid #1 has been oxidized to an alpha, beta unsaturated carboxylic acid functionality. This major cyclosporine metabolite is inactive in a phytohemagglutinin stimulated lymphocyte proliferation assay.     

Endothelium Dependent and Independent Relaxation of Aortic Rings from Watanabe Heritable Hyperlipidemic Rabbits after Exposure to a Free Radical Generating System

The effects of the xanthine oxidase/hypoxanthine free radical generating system on endothelium dependent and independent relaxation were compared in aortic rings from New Zealand white rabbits and heterozygous Watanabe heritable hyperlipidemic (WHHL) rabbits with mild atherosclerosis. Studies were carried out in young (3 months) and mature (18 months) animals. Plasma cholesterol levels were significantly higher in both 3 and 18 month WHHL animals. Endothelium independent relaxation to SNP did not differ between groups. However, the attenuation of relaxation to carbachol after xanthine oxidase/hypoxanthine treatment tended to be less in WHHL. This reached significance at 18 but not 3 months. We propose that this could be due to increases in levels of endogenous scavenger enzymes in these WHHL rabbits.     

Increased proton leak and SOD2 expression in myotubes from obese non-diabetic subjects with a family history of type 2 diabetes

Muscle insulin resistance is linked to oxidative stress and decreased mitochondrial function. However, the exact cause of muscle insulin resistance is still unknown. Since offspring of patients with type 2 diabetes mellitus (T2DM) are susceptible to developing insulin resistance, they are ideal for studying the early development of insulin resistance. By using primary muscle cells derived from obese non-diabetic subjects with (FH +) or without (FH ?) a family history of T2DM, we aimed to better understand the link between mitochondrial function, oxidative stress, and muscle insulin resistance. Insulin-stimulated glucose uptake and glycogen synthesis were normal in FH + myotubes. Resting oxygen consumption rate was not different between groups. However, proton leak was higher in FH + myotubes. This was associated with lower ATP content and decreased mitochondrial membrane potential in FH + myotubes. Surprisingly, mtDNA content was higher in FH + myotubes. Oxidative stress level was not different between FH + and FH ? groups. Reactive oxygen species content was lower in FH + myotubes when differentiated in high glucose/insulin (25 mM/150 pM), which could be due to higher oxidative stress defenses (SOD2 expression and uncoupled respiration). The increased antioxidant defenses and mtDNA content in FH + myotubes suggest the existence of compensatory mechanisms, which may provisionally prevent the development of insulin resistance.     

SIRT1 Catalytic Activity Has Little Effect on Tumor Formation and Metastases in a Mouse Model of Breast Cancer

The protein deacetylase SIRT1 has been implicated in the regulation of a large number of cellular processes that are thought to be required for cancer initiation and progression. There are conflicting data that make it unclear whether Sirt1 functions as an oncogene or tumor suppressor. To assess the effect of SIRT1 on the emergence and progression of mammary tumors, we crossed mice that harbor a point mutation that abolishes SIRT1 catalytic activity with mice carrying the polyoma middle T transgene driven by the murine mammary tumor virus promoter (MMTV-PyMT). The absence of SIRT1 catalytic activity neither accelerated nor blocked the formation of tumors and metastases in this model. There was a lag in tumor latency that modestly extended survival in Sirt1 mutant mice that we attribute to a delay in mammary gland development and not to a direct effect of SIRT1 on carcinogenesis. These results are consistent with previous evidence suggesting that Sirt1 is not a tumor promoter or a tumor suppressor.     

Salmonella in raccoons (Procyon lotor) in southern Ontario, Canada

Numerous serotypes of Salmonella have been detected in a variety of wild animals, including raccoons (Procyon lotor). Raccoons are common, mid-size omnivores that live in close association with people in urban and rural areas in Ontario. Although raccoons are known to shed Salmonella, little is known about their potential long-term role in maintaining Salmonella infections. We sampled feces from raccoons in three areas of Ontario: one primarily urban site around Niagara, one primarily rural site north of Guelph, and the grounds of the Toronto Zoo, in 2007 to identify which serotypes of Salmonella were commonly shed by raccoons in southern Ontario. In addition, we conducted a longitudinal study at the Toronto Zoo site to determine if raccoons remain persistently infected with Salmonella. Salmonella was found in 45% of samples. The prevalence of Salmonella in raccoon feces ranged from 27% at the rural site to 65% at the urban site. We detected 16 serotypes of Salmonella in 83 positive samples. The most common serotype detected in raccoons from the rural and zoo sites was Salmonella enterica serotype Typhimurium, whereas Salmonella Newport was detected most commonly in the urban site. Only one raccoon of 11 that were captured in four or more consecutive trapping sessions shed the same Salmonella serotype for two consecutive months, suggesting that raccoons regularly acquire new Salmonella serotypes from the environment.     

Survival of adult Common Guillemots Uria aalge at three Scottish colonies

We present the results of a mark–recapture analysis of survival rates of Common Guillemots with at least one year's breeding experience for birds from three Scottish colonies. Estimates of overall survival of adults from the Isle of May (North Sea), Canna and Colonsay (both off northwest Scotland) from an analysis combining data from the three colonies were 94.8% (se = ±0.6), 92.4% (se = ±0.9) and 96.7% (se = ±0.6) respectively. The rates from Canna (but not Colonsay) differed significantly from those of guillemots on the Isle of May. The Canna and Colonsay figures are the first survival estimates for guillemots in northwest Scotland, an area of high conservation concern for this species. Low input, long-term studies of seabirds can produce important data on adult survival.     

Optimising predictive modelling of Ross River virus using meteorological variables

BackgroundStatistical models are regularly used in the forecasting and surveillance of infectious diseases to guide public health. Variable selection assists in determining factors associated with disease transmission, however, often overlooked in this process is the evaluation and suitability of the statistical model used in forecasting disease transmission and outbreaks. Here we aim to evaluate several modelling methods to optimise predictive modelling of Ross River virus (RRV) disease notifications and outbreaks in epidemiological important regions of Victoria and Western Australia.Methodology/Principal findingsWe developed several statistical methods using meteorological and RRV surveillance data from July 2000 until June 2018 in Victoria and from July 1991 until June 2018 in Western Australia. Models were developed for 11 Local Government Areas (LGAs) in Victoria and seven LGAs in Western Australia. We found generalised additive models and generalised boosted regression models, and generalised additive models and negative binomial models to be the best fit models when predicting RRV outbreaks and notifications, respectively. No association was found with a model’s ability to predict RRV notifications in LGAs with greater RRV activity, or for outbreak predictions to have a higher accuracy in LGAs with greater RRV notifications. Moreover, we assessed the use of factor analysis to generate independent variables used in predictive modelling. In the majority of LGAs, this method did not result in better model predictive performance.Conclusions/SignificanceWe demonstrate that models which are developed and used for predicting disease notifications may not be suitable for predicting disease outbreaks, or vice versa. Furthermore, poor predictive performance in modelling disease transmissions may be the result of inappropriate model selection methods. Our findings provide approaches and methods to facilitate the selection of the best fit statistical model for predicting mosquito-borne disease notifications and outbreaks used for disease surveillance.     

Experimental test of connector rotation during DNA packaging into bacteriophage phi29 capsids

The bacteriophage phi29 generates large forces to compact its double-stranded DNA genome into a protein capsid by means of a portal motor complex. Several mechanical models for the generation of these high forces by the motor complex predict coupling of DNA translocation to rotation of the head-tail connector dodecamer. Putative connector rotation is investigated here by combining the methods of single-molecule force spectroscopy with polarization-sensitive single-molecule fluorescence. In our experiment, we observe motor function in several packaging complexes in parallel using video microscopy of bead position in a magnetic trap. At the same time, we follow the orientation of single fluorophores attached to the portal motor connector. From our data, we can exclude connector rotation with greater than 99% probability and therefore answer a long-standing mechanistic question.