Inbreeding trends and genetic diversity in purebred sheep populations.

Monitoring the rate of change in inbreeding and genetic diversity within a population is important to guide breeding programmes. Such interest stems from the impact of loss in genetic diversity on sustainable genetic gain but also the impact on performance (i.e. inbreeding depression). The objective of the present study was to evaluate trends in inbreeding and genetic diversity in 43 066 Belclare, 120 753 Charollais, 22 652 Galway, 78 925 Suffolk, 187 395 Texel, and 19 821 Vendeen purebred sheep. The effective population size for each of the six breeds was between 116.0 (Belclare population) and 314.8 (Charollais population). The Charollais population was the most genetically diverse with the greatest number of effective founders, effective ancestors, and effective founder genomes; conversely, the Belclare was the least genetically diverse population with the fewest number of effective founders, effective ancestors, and effective founder genomes for each of the six breeds investigated. Overall, the effective population sizes and the total genetic diversity within each of the six breeds were above the minimum thresholds generally considered to be required for the long-term viability of a population.

Overturning circulation, nutrient limitation, and warming in the Glacial North Pacific.

Although the Pacific Ocean is a major reservoir of heat and CO2, and thus an important component of the global climate system, its circulation under different climatic conditions is poorly understood. Here, we present evidence that during the Last Glacial Maximum (LGM), the North Pacific was better ventilated at intermediate depths and had surface waters with lower nutrients, higher salinity, and warmer temperatures compared to today. Modeling shows that this pattern is well explained by enhanced Pacific meridional overturning circulation (PMOC), which brings warm, salty, and nutrient-poor subtropical waters to high latitudes. Enhanced PMOC at the LGM would have lowered atmospheric CO2-in part through synergy with the Southern Ocean-and supported an equable regional climate, which may have aided human habitability in Beringia, and migration from Asia to North America.

Toxoplasma gondii in Ireland: seroprevalence and novel molecular detection method in sheep, pigs, deer and chickens.

Toxoplasma gondii is among the most studied parasites worldwide but there is not much information about it published in Ireland. The objectives of this study were to determine the seroprevalence of T. gondii in sheep, pigs, deer and chickens and the molecular detection of T. gondii DNA in muscle tissue. Serum samples were collected from these species at the time of slaughter at Irish abattoirs during 2007 and tested for anti-T. gondii antibodies using a commercial semi-quantitative latex agglutination test. Antibodies (titre ≥1 : 64) were found in 36% (105/292) sheep, 4.7% (15/317) pigs and 6.6% (23/348) deer. In chickens, 18% (65/364) had antibody titres, ranging between 1 : 5 and 1 : 1024. Significant (P ≤ 0.05) age-related differences in seroprevalence were found in adult sheep (58.1%) and pigs (23.1%). Significant gender differences in seroprevalence was also found in sheep with more females (43%) than males (22.4%) being positive. However, when adjusted for age through logistic regression gender was no longer significant. Seroprevalence was also evaluated on farm locations grouped to NUTS level 3, but the prevalence was too low to draw any statistical conclusions. Using a nested PCR, the presence of T. gondii DNA was detected in diaphragm samples from 3.6% (3/83) sheep, 13.0% (3/23) pig and 4.2% (3/71) deer. Meat digestion liquids from a Trichinella spp. survey in pigs were also used for the first time to detect T. gondii. Toxoplasma gondii DNA was detected in 50% (10/20) of pooled samples. This is the first in depth study of T. gondii seroprevalence in animals in Ireland and a novel method, using digestion liquid from pooled diaphragm samples, for PCR detection in pigs is described.

Real-time three-dimensional myocardial contrast echocardiography: is it clinically feasible?

AIMS: Real-time 3D echocardiography (RT3DE) and 2D low mechanical index (LMI), contrast specific, myocardial perfusion imaging are now both accepted techniques. We evaluated the feasibility of an RT3DE LMI implementation in unselected patients. METHODS AND RESULTS: Forty-six patients undergoing contrast enhanced dobutamine stress echo were imaged with novel 3D LMI power modulation software. All patients underwent contrast enhanced 2D and RT3DE acquisitions, in left ventricular opacification (LVO), and LMI perfusion modes. The data sets were evaluated segmentally for wall motion (WM) and myocardial contrast enhancement. Of the 736 evaluated segments, WM could be assessed in 726 (98.6%) of the 2D and 708 (96.2%) 3D segments (P = 0.007). Perfusion could be assessed in 721 (98%) of 2D and 701 (95.2%) of 3D segments (P = 0.006). Six hundred and sixty-one segments had normal WM and thickening in 2D and of these RT3DE demonstrated normal myocardial opacification in 77.2% of basal, 85% of mid, and 91.8% of apical segments. Thirty-four segments were akinetic, with no evidence of perfusion in 2D, and of these RT3DE revealed a perfusion defect in 31 (91%, P = NS). CONCLUSION: LMI RT3DE evaluation of myocardial perfusion is feasible in most segments. It has the potential to accurately locate and possibly quantify perfusion defects.

Salmonella surveillance in raw and cooked meat and meat products in the Republic of Ireland from 2002 to 2004.

The food industry, under the regulation of the Department of Agriculture and Food (DAF) in the Republic of Ireland, is required to undertake all microbiological testing in relation to zoonoses control, in laboratories approved by DAF. These laboratories submit a monthly report of all tests undertaken, together with all presumptive Salmonella isolates for confirmation, typing and storage to the Central Veterinary Research Laboratory (CVRL). Details of Salmonella tests on 110,229 raw and 25,189 cooked meat samples from 25 laboratories were recorded over the 3-year period 2002-2004. Salmonella spp. were isolated from 1.0% of the 110,229 raw meat samples and 0.1% of the 25,189 cooked meat samples tested. The percentage of raw meat samples contaminated with Salmonella decreased over the three-year period from 1.2% to 0.9%. There was no seasonal trend in the isolation of Salmonella from any of the meats or meat products. Recoveries of the organism were highest for turkey and chicken meats at 3.1% and 2.8%, respectively, followed by porcine meats at 2.1%. The recoveries were much lower for ovine meats and meat products at 0.2% and bovine meat and meat products at 0.16%.

Ultraharmonic myocardial contrast imaging: in vivo experimental and clinical data from a novel technique.

Myocardial contrast echocardiography (MCE) with high-mechanical-index (MI), triggered harmonic imaging is the best-established technique to date for the assessment of myocardial perfusion. A high signal-to-noise ratio, which is significantly influenced by precontrast tissue signals, is an important prerequisite. Our goal was to evaluate the efficacy of ultraharmonic MCE, a technique that rejects tissue signals by receiving signals beyond the second but below the third harmonic. Imaging was performed in 6 closed-chest dogs and in 15 healthy volunteers (11 of whom also had dipyridamole stress). Analyses of videointensity (VI) confirmed uniformly low precontrast tissue VI, a significant increase of postcontrast VI (before and after dipyridamole), and a significant decrease in VI after microbubble destruction. We conclude that ultraharmonic MCE produces low precontrast tissue signals, thus optimizing postcontrast myocardial opacification, and exhibits efficient microbubble destruction with use of multiple-frame triggering. Thus this new technique opens up a new possibility of further optimizing coronary microcirculation imaging with microbubbles.

Cationic lipid structure and formulation considerations for optimal gene transfection of the lung.

Enhanced gene transduction to the lung using cationic lipids could be attained through optimization of the structure of the lipids and the formulation of the cationic lipid:plasmid DNA (pDNA) complexes. We have expanded on our earlier observation of the importance of the structural orientation of the cationic lipid headgroup. Through the synthesis of a number of matched pairs of cationic lipids differing only in the configuration of their headgroup, we confirmed that those harboring a T-shape headgroup are more active than their linear counterparts, at least when tested in the lungs of BALB/c mice. Additionally, we demonstrated that not only are the structural considerations of these cationic lipids important, but also their protonation state, the free base being invariably more active than its salt counterpart. The salt forms of cationic lipids bound pDNA with greater avidity, which may have affected their subsequent intracellular dissolution and transit of the pDNA to the nucleus. Inclusion of a number of frequently used solutes in the vehicle severely inhibited the gene transfection activity of the cationic lipids. The selection of neutral co-lipids was also an important factor for overall transfection activity of the formulation, with significant gains in transfection activity realized when diphytanoylphosphatidylethanolamine or dilinoleoylphosphatidylethanolamine were used in lieu of dioleoylphosphatidylethanolamine. Finally, we showed that a transacylation reaction could occur between the cationic lipid and neutral co-lipid which reduced the transfection activity of the complexes. It is the hope that as our understanding of the many factors that influence the activity of these cationic lipid:pDNA complexes improves, formulations with much greater potency can be realized for use in the treatment of pulmonary diseases.

Increased duration of transgene expression in the lung with plasmid DNA vectors harboring adenovirus E4 open reading frame 3.

For gene therapy to be effective in the treatment of chronic diseases, plasmid DNA (pDNA) vectors that provide persistent expression of therapeutic levels of the transgene product are desirable. Studies in the lung with adenovirus vectors showed that products of the adenovirus E4 region can act both in cis and in trans to increase the duration of expression when transcription of the transgene was under the control of the human cytomegalovirus (CMV) promoter. To determine if these E4-encoded proteins could also effect greater persistence of expression from a nonviral vector, a complex composed of cationic lipid GL-67, a CMV promoter plasmid (pCF1-CAT), and an E4-containing adenovirus vector (Ad2/betagal-4) was instilled into the lungs of BALB/c nu/nu mice. Significant increases in the duration of transgene expression were observed for up to 10 weeks postinstillation compared with expression from mice instilled with control complexes containing an adenovirus vector deleted of most of E4 (Ad2/betagal-2). This effect could also be observed in immunodeficient NIH-rnu rats as well as in immunocompetent BALB/c mice. Studies with CMV promoter mutants indicated that a region proximal to the promoter was necessary for the E4-mediated increase in longevity of expression. In addition to the CMV promoter, a CMV enhancer-human mucin I (MUC-I) hybrid promoter also responded to these E4-encoded proteins with increased persistence of transgene expression, but a human interleukin 8 (IL-8) promoter did not. Ad2/betagal-4 could be replaced by a pDNA vector expressing only the E4 region, indicating that products of the E4 region alone were sufficient in the absence of expression from the rest of the adenovirus genome. Further analysis indicated that the protein encoded by open reading frame 3 (ORF3) alone was sufficient for conferring the increase in persistence of expression. These data indicate that expression of a single protein from the adenovirus genome can significantly improve the duration of transgene expression from pDNA vectors, and increases the feasibility of using nonviral vectors for the treatment of chronic diseases.

Use of harmonic imaging without echocardiographic contrast to improve two-dimensional image quality.

The aim of this study was to determine whether harmonic imaging (HI) improves endocardial visualization during 2-dimensional echocardiography without echocardiographic contrast. HI differs from fundamental imaging (FI) by transmitting ultrasound at one frequency and receiving at twice the transmitted frequency. This technique has been used in conjunction with contrast echocardiography to enhance myocardial contrast visualization. HI and FI were sequentially performed in 20 patients. Images were digitally stored and subsequently reviewed by 2 observers for the quality of endocardial visualization. In addition, acoustic quantification was performed in both FI and HI modes and endocardial tracking qualitatively judged. HI was compared with FI during dobutamine stress echocardiography in 17 patients who were imaged at baseline and peak stress. Overall, the harmonic images had less clutter and better myocardial blood contrast. Individual segments were better visualized with HI in 30% to 73% of cases. The acoustic quantification endocardial tracking was rated better with HI in 67% of short-axis views and in 58% of apical 4-chamber views. During dobutamine stress testing the overall number of interpretable segments improved from 64% for FI to 84% with HI. Many segments traditionally difficult to image were improved with HI. HI without the use of contrast agents improved endocardial visualization during routine 2-dimensional echocardiography. This improved endocardial visualization led to better endocardial tracking with acoustic quantification and to more segments being clinically interpretable during dobutamine stress testing.

Increased ultrasound contrast and decreased microbubble destruction rates with triggered ultrasound imaging.

Although transient myocardial contrast imaging has been able to produce visually evident myocardial contrast in animals and humans with very low intravenous doses of perfluorocarbon-exposed sonicated dextrose albumin (PESDA) microbubbles, the mechanism for improved contrast remains unclear. In this study we devised a flow chamber that measured the concentration of PESDA microbubbles that remained after exposure to diagnostic ultrasound pressures of 0.9 to 1.9 MPa and frequencies of 2.0, 2.5, and 3.5 MHz (first and second harmonic for 2.0 MHz), which were delivered at either 30 Hz (frames per second), 0.5 to 1.0 Hz, or without any ultrasound transmission. The videointensity within the flow chamber was also measured at 0, 20, 40, and 100 ml/min flow rates with the flow loop closed (i.e., constant microbubble concentration) with both triggered (0.5 to 1.0 Hz) and conventional (30 Hz) frame rates. The effluent microbubble concentration was significantly larger when PESDA was exposed to either no ultrasound or 0.5 to 1.0 Hz ultrasound. Furthermore, the videointensity of a constant number of microbubbles was significantly greater with 0.5 to 1.0 Hz (triggered) compared with 30 Hz (conventional) frame rates at each transmit frequency. The greatest difference was noted with the lower 2.0 MHz transmit frequency and the 20 ml/min flow rate, especially when a second harmonic receiving frequency was used. We conclude that the mechanism for improved contrast with triggered ultrasound imaging is because of both less microbubble destruction and increased videointensity from a constant number of microbubbles. Lower transducer frequencies and lower flow rates result in the greatest improvement in videointensity with triggered ultrasound transmission.