Procedures for cryogenic X-ray ptychographic imaging of biological samples.

Biological sample-preparation procedures have been developed for imaging human chromosomes under cryogenic conditions. A new experimental setup, developed for imaging frozen samples using beamline I13 at Diamond Light Source, is described. This manuscript describes the equipment and experimental procedures as well as the authors' first ptychographic reconstructions using X-rays.

A high-flux high-order harmonic source.

We develop and implement an experimental strategy for the generation of high-energy high-order harmonics (HHG) in gases for studies of nonlinear processes in the soft x-ray region. We generate high-order harmonics by focusing a high energy Ti:Sapphire laser into a gas cell filled with argon or neon. The energy per pulse is optimized by an automated control of the multiple parameters that influence the generation process. This optimization procedure allows us to obtain energies per pulse and harmonic order as high as 200 nJ in argon and 20 nJ in neon, with good spatial properties, using a loose focusing geometry (f#≈400) and a 20 mm long medium. We also theoretically examine the macroscopic conditions for absorption-limited conversion efficiency and optimization of the HHG pulse energy for high-energy laser systems.

Digital in-line holography on amplitude and phase objects prepared with electron beam lithography.

We report on the fabrication and characterization of amplitude and phase samples consisting of well defined Au or Al features formed on ultrathin silicon nitride membranes. The samples were manufactured using electron beam lithography, metallization and a lift-off technique, which allow precise lateral control and thickness of the metal features. The fabricated specimens were evaluated by conventional microscopy, atomic force microscopy and with the digital in-line holography set-up at the Lund Laser Centre. The latter uses high-order harmonic generation as a light source, and is capable of recovering both the shape and phase shifting properties of the samples. We report on the details of the sample production and on the imaging tests with the holography set-up.

Photoemission electron microscopy using extreme ultraviolet attosecond pulse trains.

We report the first experiments carried out on a new imaging setup, which combines the high spatial resolution of a photoemission electron microscope (PEEM) with the temporal resolution of extreme ultraviolet (XUV) attosecond pulse trains. The very short pulses were provided by high-harmonic generation and used to illuminate lithographic structures and Au nanoparticles, which, in turn, were imaged with a PEEM resolving features below 300 nm. We argue that the spatial resolution is limited by the lack of electron energy filtering in this particular demonstration experiment. Problems with extensive space charge effects, which can occur due to the low probe pulse repetition rate and extremely short duration, are solved by reducing peak intensity while maintaining a sufficient average intensity to allow imaging. Finally, a powerful femtosecond infrared (IR) beam was combined with the XUV beam in a pump-probe setup where delays could be varied from subfemtoseconds to picoseconds. The IR pump beam could induce multiphoton electron emission in resonant features on the surface. The interaction between the electrons emitted by the pump and probe pulses could be observed.

A low-protein diet exacerbates postprandial chylomicron concentration in moderately dyslipidaemic subjects in comparison to a lean red meat protein-enriched diet.

OBJECTIVE: To investigate whether altering energy intake as red meat protein or carbohydrate modifies chylomicron homeostasis and postprandial lipaemia. DESIGN: Randomized single-blind dietary intervention trial. SETTING: School of Public Health, Division of Health Science, Curtin University, Perth, Western Australia. SUBJECTS: A total of 20 moderately hypertriglyceridaemic but otherwise healthy subjects were recruited and completed the study. INTERVENTION: Participants consumed an isocaloric weight maintenance diet low in protein (14, 53 and 30% of energy as protein, carbohydrate and fat, respectively) or high in protein (25, 35 and 30% energy as protein, carbohydrate and fat) for a period of 6 weeks. Fasting plasma lipids and postprandial lipoprotein studies (triglyceride and apolipoprotein B48) following an oral fat challenge were carried out at the start and conclusion of the dietary intervention period. RESULTS: Consumption of the low- or high-protein diet had no significant effect on fasting plasma or postprandial lipaemia, the latter determined as the incremental area under the triglyceride curve following a fat challenge. However, subjects who consumed a low-protein diet for 6 weeks had a substantially exaggerated postprandial chylomicron response, indicated as the area under the apo B48 curve following a fat challenge. The change in postprandial chylomicron kinetics could not be explained by changes in insulin sensitivity, which appeared to be similar before and after intervention with either diet. CONCLUSIONS: Daily moderate consumption of a lean red meat protein-enriched diet attenuates postprandial chylomicronaemia in response to ingestion of a fatty meal.

Effect of source of cattle and distance transported to a commercial slaughter facility on carcass bruises in mature beef cows.

OBJECTIVE: To determine the effect of source of cattle and distance cattle were transported to a commercial slaughter facility on prevalence and severity of carcass bruises in mature beef cows. DESIGN: Epidemiologic investigation. SAMPLE POPULATION: Carcasses from 3,955 beef cows from 11 states assembled in 89 procurement lots. PROCEDURE: Each carcass was scored for bruise severity and location. Source of cattle was categorized. Distance from source to slaughter facility was determined. An ANCOVA was used to determine effects of variables on carcass bruises. RESULTS: Mature beef cows marketed through livestock auctions conducting first-point testing for brucellosis, especially when transported longer distances (> 325 km) to slaughter facilities, had a greater number and severity of carcass bruises than cows originating from ranches or livestock auctions not conducting first-point testing. There was an increase in number of rib bruises in cows originating from livestock auctions not conducting first-point testing. Prevalence and severity of bruises were not significantly affected by transportation distance between cows originating from auctions not conducting first-point testing and cows originating from ranches. CLINICAL IMPLICATIONS: A strong association existed between handling for brucellosis testing prior to sale for slaughter and distance transported to slaughter facility with carcass bruises in mature beef cows. Bruises are major quality defects that decrease carcass value and slaughter-cow prices. Repeated handling and restraint as well as long-distance transport are issues to consider regarding the responsibility of the livestock industry to provide for the safety and well-being of cattle sold for slaughter.

Persistent pulmonary hypertension of the neonate and asymmetric growth restriction.

OBJECTIVE: To evaluate the possible associations between persistent pulmonary hypertension of the neonate, need for extra-corporeal membranous oxygenation, small for gestational age (SGA), and low ponderal index for gestational age in infants with persistent pulmonary hypertension of the neonate and in matched controls. METHODS: Eighty-six infants with persistent pulmonary hypertension of the neonate delivered from 1991 to 1994 at our hospital were matched with 430 contemporaneous control singleton neonates. Birth weight and ponderal indices (100 x weight/length3) less than the tenth percentile for gestational age and gender were defined as SGA and low ponderal index, respectively. We assessed associations between these markers, the presence of persistent pulmonary hypertension of the neonate, and the need for extracorporeal membranous oxygenation. RESULTS: Low ponderal index was associated with persistent pulmonary hypertension of the neonate (odds ratio [OR] 5.4), whereas SGA was not. Low ponderal index (OR 4.0) was an independent correlate of persistent pulmonary hypertension of the neonate after adjustment with logistic regression for 5-minute Apgar scores less than 7, umbilical arterial pH less than 7.10, and presence of meconium. Low ponderal index was associated with need for extracorporeal membranous oxygenation in neonates with persistent pulmonary hypertension (P < .001). CONCLUSION: Fetal developmental events may significantly affect neonatal pulmonary status. Diminished neonatal nutritional status, as measured by low ponderal index for gestational age, is associated with increased risk of persistent pulmonary hypertension of the neonate and severity of the disease process.

A comparison of two serological methods for detecting the immune response after rabies vaccination in dogs and cats being exported to rabies-free areas.

Levels of rabies virus neutralizing antibody in sera from dogs and cats were titrated to endpoint by the Rapid Fluorescent Focus Inhibition Test (RFFIT) and retested by the RFFIT and the Fluorescent Antibody Virus Neutralization test (FAVN). The two tests were compared for their ability to detect the 0.5 international units/ml (I.U.) of antibody required by the World Health Organization and the Office International des Epizooties as the minimum response for proof of rabies immunization. No difference was observed in sensitivity or specificity for either method in tests of 168 sera from unvaccinated animals or 70 sera from vaccinated animals with high levels of neutralizing antibody (an initial RFFIT titre of > or = 1.0 I.U.). Test to test variation occurred for results obtained by both RFFIT and FAVN for 95 sera from vaccinated animals with low to moderate levels of neutralizing antibody (RFFIT titre < 1.0 I.U.). No significant differences were detected for the 95 sera in the frequency for one methodology more often than the other to have a positive response (> or = 0.5 I.U.), nor were significant differences detected for the symmetry (P = 0.43) or the marginal homogeneity (P = 0.39) of results obtained by the two methods. Both methods can adequately identity unvaccinated animals, but false positive and false negative results are possible for either method when a single test is used to measure the antibody response of low-responding vaccinated animals. Nucleotide sequence analysis identified several amino acid differences in stocks of the challenge rabies virus from different laboratories. The small differences in neutralizing antibody titre that may result from mutations in the challenge virus are not important for evaluating immunity induced by vaccines which are themselves prepared from a variety of different rabies virus strains, but differences in the challenge virus, rather than differences in methodology, may account for at least some of the discrepant results reported in inter-laboratory surveys. Comparative studies of serological methods for measuring rabies antibodies should use well-characterized unpassaged virus stocks obtained from a single reference laboratory.

Estimating the maximum effective dose in a quantitative dose-response experiment.

A simulation study was conducted to compare several procedures for estimating the maximum effective dose in a quantitative dose-response experiment. Using four equally spaced dose levels, data were generated from four different model types: the quadratic growth curve, the Mitcherlich growth curve, the linear-linear plateau spline model, and the quadratic-linear plateau spline model. Each model type was parameterized to create three different model ranges, and for each range, data were generated from populations with three different standard deviations. The existence of unique dose-response curves is assumed; thus, all the procedures compared in this paper require that the data have been modeled by a polynomial or nonlinear regression model. An attempt was made to fit each generated data set with each of the four model types. Maximum effective dose estimation procedures were applied to a data set only when the data were adequately described by a given model. The stimulation indicated that the estimate of the maximum effective dose is influenced more by the choice of model than by the method of estimation. Because of the consistently low estimates produced when the data were modeled by the linear-linear plateau spline, this model is not recommended for use an maximum effective dose estimation experiments. The simulation also demonstrated that the design failed to provide sufficient information about the form of the dose-response curve. Designs with more than four dose levels should be considered.

Sensory evaluation of beef-flavor-intensity, tenderness, and juiciness among major muscles.

Twelve muscles from eight USDA Select/Choice grade steers were evaluated for beef-flavor intensity, tenderness, and juiciness. The biceps femoris, psoas major, gluteus medius, semimembranosus, and triceps brachii were similar in beef-flavor-intensity (P > 0·05) and were ranked as the most intensely flavored of all muscles. The rectus femoris, longissimus lumborum, serratus ventralis, infraspinatus, semitendinosus, pectoralis profundus, and supraspinatus generally were less intense in beef-flavor than the other muscles and were ranked from highest to lowest intensity in that order. The psoas major was the most tender (P > 0·05) followed by the infraspinatus, longissimus lumborum, and rectus femoris, which were similar (P > 0·05). Generally, muscles from the chuck and loin were juicier than those from the round.

Comparisons of the effect of electrical stimulation methods on postmortem pH decline in beef muscle.

Results of four electrical stimulation (ES) studies were summarized to demonstrate the impact of different ES parameters on pH decline patterns in postmortem M. longissimus thoracis et lumborum. Postmortem pH decline was modeled as a non-linear function of time, and estimates of minimum obtainable pH, pH decline rates, and time to reach pH 6·0 were compared for each study. The decline model for study 4 (AC, 60 Hz, 50 V; 5 min post mortem) had a larger (P < 0·05) estimate for decline rate than that for study 1 (AC, 60 Hz, 400 V; 1 h post mortem) and the control (non-stimulated) data. The model estimate of time to pH 6·0 (0·56 h) for study 4 was the shortest (P < 0·05) for all treatments. Different ES parameters produce different pH decline patterns post mortem and, therefore, may impact product quality and fabrication and chilling protocols adopted in fresh beef processing.

Longevity of rabies antibody titre in recipients of human diploid cell rabies vaccine.

Sera from individuals across the USA and Peace Corps Volunteers (PCV) were assayed for neutralizing antibody to human diploid cell rabies vaccine (HDCV). A pairwise comparison between intramuscular (i.m.) and intradermal (i.d.) administration, PCV and non-PCV, revealed statistically significant differences between i.m. and i.d. for PCV and non-PCV, and between PCV and non-PCV for i.d. and i.m. administration. Survival analysis and frequency tables employing chi 2 tests determined that the percentage probability of maintaining an adequate titre from 1.5-2.0 years after primary vaccination was 99% in non-PCV i.m., 93% in non-PCV i.d., 88% in PCV i.m., and 64% in PCV i.d.

Landing pattern of stable flies (Diptera: Muscidae) on the alsynite cylinder trap: effect of wind speed and direction.

The distribution of stable flies, Stomoxys calcitrans (L.), caught on adhesive-coated Alsynite cylinder traps indicated that a significantly higher proportion of flies landed on the side most protected from the wind, and that flies were distributed equally on both sides of the traps bisected by the direction of the prevailing wind, and that the proportion of trapped flies decreased significantly with height on the trap. The landing pattern of house flies, Musca domestica L., appeared to differ from that of stable flies. As wind speed increased, the proportion of stable flies landing on the lower and downwind parts of the trap increased.

Autoregressive errors with a repeated measures design in clinical trials.

Repeated measures designs are common in clinical trials where recordings on a patient are made repeatedly over time. A basic assumption for the analysis of repeated measures designs is that a common correlation structure between observations for a given patient exists. When several observations are made for each patient this assumption may be tenuous at best. This article presents a pragmatic approach for combining a repeated measures design with a first-order autoregressive error component. A method of filtering the observed data to account for the autoregressive structure of the errors is considered. The effect on the analysis of variance results after extraction of the autoregressive component is seen by comparison of ANOVA summaries.