Estimating the impact of various pathway parameters on tenderness, flavour and juiciness of pork using Monte Carlo simulation methods.

Monte Carlo simulation was investigated as a potential methodology to estimate sensory tenderness, flavour and juiciness scores of pork following the implementation of key pathway interventions known to influence eating quality. Correction factors were established using mean data from published studies investigating key production, processing and cooking parameters. Probability distributions of correction factors were developed for single pathway parameters only, due to lack of interaction data. Except for moisture infusion, ageing period, aitchbone hanging and cooking pork to an internal temperature of >74°C, only small shifts in the mean of the probability distributions of correction factors were observed for the majority of pathway parameters investigated in this study. Output distributions of sensory scores, generated from Monte Carlo simulations of input distributions of correction factors and for individual pigs, indicated that this methodology may be useful in estimating both the shift and variability in pork eating traits when different pathway interventions are applied.

Can deuterium stable isotope values be used to assign the geographic origin of an auxiliary hoverfly in south-western France?

Deuterium δD isotopic analysis is increasingly being used to trace wildlife movement, and undoubtedly has much to offer in this respect, but questions still remain as to the feasibility and practicality of the method in ecology. Here we report our attempt to determine the geographic origin of an auxiliary hoverfly, Episyrphus balteatus, in south-western France. We used quantile regression to calculate the minimum separation distance, based on the International Atomic Energy Agency/World Meteorological Organization (IAEA/WMO) data, at which two insects could be said to originate from different latitudes with a given degree of confidence. We collected larvae in spring 2007 and 2009 to obtain the δD signal of indigenous hoverflies and we trapped adults during one complete year (from Dec. 2006 to Nov. 2007). The smallest separation distance calculated was about 1400 km in western Europe. Our results revealed greater variability in δD of adults in autumn than in spring. From this we infer an autumnal migration. Because of the presence of mountains and the Mediterranean Sea, the δD gradient in precipitation in western Europe is less clear than on the American continent, where it has been used successfully to infer geographical origins of animals under certain conditions. Despite the complications encountered in Europe, the minimum separation distance model proved a useful first step to obtain a first range of possible origins of E. balteatus and the application of the model to other arthropod species in Europe warrants investigation.

Estimators of annual probability of infection for quantitative microbial risk assessment.

Four estimators of annual infection probability were compared pertinent to Quantitative Microbial Risk Analysis (QMRA). A stochastic model, the Gold Standard, was used as the benchmark. It is a product of independent daily infection probabilities which in turn are based on daily doses. An alternative and commonly-used estimator, here referred to as the Naïve, assumes a single daily infection probability from a single value of daily dose. The typical use of this estimator in stochastic QMRA involves the generation of a distribution of annual infection probabilities, but since each of these is based on a single realisation of the dose distribution, the resultant annual infection probability distribution simply represents a set of inaccurate estimates. While the medians of both distributions were within an order of magnitude for our test scenario, the 95th percentiles, which are sometimes used in QMRA as conservative estimates of risk, differed by around one order of magnitude. The other two estimators examined, the Geometric and Arithmetic, were closely related to the Naïve and use the same equation, and both proved to be poor estimators. Lastly, this paper proposes a simple adjustment to the Gold Standard equation accommodating periodic infection probabilities when the daily infection probabilities are unknown.

Validation of fixed sample size plans for monitoring lepidopteran pests of Brassica oleracea crops in North Korea.

The combined action of two lepidoteran pests, Plutella xylostella L. (Plutellidae) and Pieris rapae L. (Pieridae),causes significant yield losses in cabbage (Brassica oleracea variety capitata) crops in the Democratic People's Republic of Korea. Integrated pest management (IPM) strategies for these cropping systems are in their infancy, and sampling plans have not yet been developed. We used statistical resampling to assess the performance of fixed sample size plans (ranging from 10 to 50 plants). First, the precision (D = SE/mean) of the plans in estimating the population mean was assessed. There was substantial variation in achieved D for all sample sizes, and sample sizes of at least 20 and 45 plants were required to achieve the acceptable precision level of D < or = 0.3 at least 50 and 75% of the time, respectively. Second, the performance of the plans in classifying the population density relative to an economic threshold (ET) was assessed. To account for the different damage potentials of the two species the ETs were defined in terms of standard insects (SIs), where 1 SI = 1 P. rapae = 5 P. xylostella larvae. The plans were implemented using different economic thresholds (ETs) for the three growth stages of the crop: precupping (1 SI/plant), cupping (0.5 SI/plant), and heading (4 SI/plant). Improvement in the classification certainty with increasing sample sizes could be seen through the increasing steepness of operating characteristic curves. Rather than prescribe a particular plan, we suggest that the results of these analyses be used to inform practitioners of the relative merits of the different sample sizes.

The Northern Ireland Public Access Defibrillation (NIPAD) study: effectiveness in urban and rural populations.

OBJECTIVE: To assess the impact of mobile automated external defibrillators (AEDs) on out-of-hospital cardiac arrests (OHCAs) in urban and rural populations. DESIGN: Prospective before and after intervention, population study. SETTING: Urban and rural areas of 160,000 each. Patients, interventions and MAIN OUTCOME MEASURES: In 2004-6 the demographics of OHCAs were assessed. In 2005-6 AEDs were deployed (29 urban, 53 rural): 335 urban first responders (FRs) and 493 rural FRs were trained in AED use and dispatched to OHCAs. Call-to-response interval (CRI), resuscitation and survival-to-discharge rates for OHCA were compared. RESULTS: In 2004 there were 163 urban OHCAs and the emergency medical services (EMS) attended 158 (ventricular fibrillation (VF) 27/158 (17.1%)). In 2005-6 there were 226 OHCAs, EMS attended 216 (VF 30/216 (13.9%)). In 2005-6 FRs were paged to 128 OHCAs (56.6%), FRs attended 88/128 (68.8%): 18/128 (14.1%) reached before the EMS. The best combined FR/EMS mean (SD) CRI in 2005-6 (5 min 56 s (4)) was better than the EMS alone in 2004 (7 min (3); p = 0.002). Survival rate was 5.1% in 2004, 1.4% in 2005-6 (p = NS). In 2004 there were 131 rural OHCAs, EMS attended 121 (VF 19/121 (15.7%)). In 2005-6 there were 122 OHCAs, EMS attended 114 (VF 19/114 (16.7%)). In 2005-6 FRs were paged to 49 OHCAs, FRs attended 42/49 (85.7%): 23/49 (46.9%) reached before the EMS. The best combined FR/EMS mean (SD) CRI in 2005-6 (9 min 22 s (6)) was better than the EMS alone in 2004 (11 min 2 s (6); p = 0.018). Survival rate was 2.5% in 2004, 3.5% in 2005-6 (p = NS). CONCLUSIONS: Despite improvement in CRI there was no impact on survival (witnessed arrest 32.8%, VF 15.6%). TRIAL REGISTRATION NUMBER: ISRCTN07286796.

The obstacles to maximising the impact of public access defibrillation: an assessment of the dispatch mechanism for out-of-hospital cardiac arrest.

OBJECTIVES: To determine the diagnostic accuracy of advanced medical priority dispatch system (AMPDS) software used to dispatch public access defibrillation first responders to out-of-hospital cardiac arrests (OHCA). DESIGN: All true OHCA events in North and West Belfast in 2004 were prospectively collated. This was achieved by a comprehensive search of all manually completed Patient Report Forms compiled by paramedics, together with autopsy reports, death certificates and medical records. The dispatch coding of all emergency calls by AMPDS software was also obtained for the same time period and region, and a comparison was made between these two datasets. SETTING: A single urban ambulance control centre in Northern Ireland. POPULATION: All 238 individuals with a presumed or actual OHCA in the North and West Belfast Health and Social Services Trust population of 138 591 (2001 Census), as defined by the Utstein Criteria. MAIN OUTCOME MEASURES: The accurate dispatch of an emergency ambulance to a true OHCA. RESULTS: The sensitivity of the dispatch mechanism for detecting OHCA was 68.9% (115/167, 95% confidence interval (CI) 61.3% to 75.8%). However, the sensitivity for arrests with ventricular fibrillation (VF) was 44.4% (12/27) with sensitivity for witnessed VF of 47.1% (8/17). The positive predictive value was 63.5% (115/181, 95% CI 56.1% to 70.6%). CONCLUSIONS: The sensitivity of this dispatch process for cardiac arrest is moderate and will constrain the effectiveness of Public Access Defibrillation (PAD) schemes which utilise it. TRIAL REGISTRATION: controlled-trials.com ISRCTN07286796.

Regional-scale models for relating land cover to basin surface-water quality using remotely sensed data in a GIS.

Plant-based management systems implementing deep-rooted, perennial vegetation have been identified as important in mitigating the spread of secondary dryland salinity due to its capacity to influence water table depth. The Glenelg Hopkins catchment is a highly modified watershed in the southwest region of Victoria, where dryland salinity management has been identified as a priority. Empirical relationships between the proportion of native vegetation and in-stream salinity were examined in the Glenelg Hopkins catchment using a linear regression approach. Whilst investigations of these relationships are not unique, this is the first comprehensive attempt to establish a link between land use and in-stream salinity in the study area. The results indicate that higher percentage land cover with native vegetation was negatively correlated with elevated in-stream salinity. This inverse correlation was consistent across the 3 years examined (1980, 1995, and 2002). Recognising the potential for erroneously inferring causal relationships, the methodology outlined here was both a time and cost-effective tool to inform management strategies at a regional scale, particularly in areas where processes may be operating at scales not easily addressed with on-site studies.

Is the risk of illness through consuming vegetables irrigated with reclaimed wastewater different for different population groups?

The use of reclaimed wastewater for irrigation of horticultural crops is commonplace in many parts of the world and is likely to increase. Concerns about risks to human health arising from such practice, especially with respect to infection with microbial pathogens, are common. Several factors need to be considered when attempting to quantify the risk posed to a population, such as the concentration of pathogens in the source water, water treatment efficiency, the volume of water coming into contact with the crop, and the die-off rate of pathogens in the environment. Another factor, which has received relatively less attention, is the amount of food consumed. Plainly, higher consumption rates place one at greater risk of becoming infected. The amount of vegetables consumed is known to vary among ethic groups. We use Quantitative Microbial Risk Assessment Modelling (QMRA) to see if certain ethnic groups are exposed to higher risks by virtue of their consumption behaviour. The results suggest that despite the disparities in consumption rates by different ethnic groups they generally all faced comparable levels of risks. We conclude by suggesting that QMRA should be used to assess the relative levels of risk faced by groups based on divisions other than ethnicity, such as those with compromised immune systems.

Evaluation of fixed sample-size plans for Plutella xylostella (Lepidoptera: Plutellidae) on broccoli crops in Australia.

Fixed sample-size plans for monitoring Plutella xylostella (L.) (Lepidoptera: Plutellidae) on broccoli and other Brassica vegetable crops are popular in Australia for their simplicity and ease of application. But the sample sizes used are often small, approximately 10-25 plants per crop, and it may be that they fail to provide sufficient information upon which to base pest control decisions. We tested the performance of seven fixed sample-size plans (10, 15, 20, 30, 35, 40, and 45 plants) by resampling a large data set on P. xylostella in commercial broccoli crops. For each sample size, enumerative and presence-absence plans were assessed. The precision of the plans was assessed in terms of the ratio of the standard error to the mean; and at least 45 and 35 samples were necessary for the enumerative and presence-absence plans, respectively, to attain the generally accepted benchmark of < or = 0.3. Sample sizes of 10-20 were highly imprecise. We also assessed the consequences of classifications based on action thresholds (ATs) of 0.2 and 0.8 larvae per plant for the enumerative case, and 0.15 and 0.45 proportion of plants of infested for the presence-absence case. Operating characteristic curves and investigations of the frequency of correct decisions suggest improvements in the performance of plans with increased sample size. In both the enumerative and presence-absence cases, the proportion of incorrect decisions was much higher for the lower of the two ATs assessed, and type II errors (i.e., failure to suggest pest control upon the AT is exceeded) generally accounted for the majority of this error. Type II errors are the most significant from a producer's standpoint. Further consideration is necessary to determine what is an acceptable type II error rate.

Scytalidium dimidiatum causing recalcitrant subcutaneous lesions produces melanin.

Scytalidium dimidiatum is a pigmented dematiaceous coelomycete that typically causes chronic superficial skin diseases and onychomycosis, as well as deeper infections, such as subcutaneous abscesses, mycetoma, and even fungemia in immunocompromised patients. A second species, Scytalidium hyalinum, has hyaline hyphae and arthroconidia and is considered by some authors to be an albino mutant of S. dimidiatum. This study aimed to confirm the presence of melanin or melanin-like compounds (which have been previously implicated in the virulence of other fungal pathogens) in S. dimidiatum from a patient with multiple subcutaneous nodules. Treatment of the hyphae and arthroconidia with proteolytic enzymes, denaturant, and concentrated hot acid yielded dark particles, which were stable free radicals, consistent with their identification as melanins. Extracted melanin particles from S. dimidiatum cultures were labeled by melanin-binding monoclonal antibodies (MAbs) from Sporothrix schenckii, Aspergillus fumigatus, and Cryptococcus neoformans. Lesional skin from the patient infected with S. dimidiatum contained fungal cells that were labeled by melanin-binding MAbs, and digestion of the tissue yielded dark particles that were also reactive. S. hyalinum was also subjected to the melanin extraction protocol, but no dark particles were yielded.

Accounting for cluster sampling in constructing enumerative sequential sampling plans.

Green's sequential sampling plan is widely used in applied entomology. Green's equation can be used to construct sampling stop charts, and a crop can then be surveyed using a simple random sampling (SRS) approach. In practice, however, crops are rarely surveyed according to SRS. Rather, some type of hierarchical design is usually used, such as cluster sampling, where sampling units form distinct groups. This article explains how to make adjustments to sampling plans that intend to use cluster sampling, a commonly used hierarchical design, rather than SRS. The methodologies are illustrated using diamondback moth, Plutella xylostella (L.), a pest of Brassica crops, as an example.

A dynamic binomial sequential sampling plan for Plutella xylostella (Lepidoptera: Plutellidae) on broccoli and cauliflower in Australia.

Binomial sequential sampling plans have been used widely for monitoring invertebrate pest populations. Such plans are typically based upon a single action threshold (AT), which represents the level of infestation that the grower is prepared to accept before using a control measure. For many cropping systems this acceptable infestation level is likely to vary, being dependent on factors such as the growth stage of the crop and the value or demands of the destination market (e.g., local or high-quality export). We developed and validated a computer-assisted plan that uses a dynamic AT. The plan has been developed for monitoring diamondback moth, Plutella xylostella (L.) on broccoli (Brassica oleracea variety botrytis L.) and cauliflower (Brassica oleracea variety botrytis L.), but the concepts and methodologies could be readily applied to other systems.

Effects of high altitude and water deprivation on arginine vasopressin release in men.

High-altitude exposure changes the distribution of body water and electrolytes. Arginine vasopressin (AVP) may influence these alterations. The purpose of this study was to examine the effect of a 24-h water deprivation trial (WDT) on AVP release after differing altitude exposures. Seven healthy males (age 22 +/- 1 yr, height 176 +/- 2 cm, mass 75.3 +/- 1.8 kg) completed three WDTs: at sea level (SL), after acute altitude exposure (2 days) to 4,300 m (AA), and after prolonged altitude exposure (20 days) to 4,300 m (PA). Body mass, standing and supine blood pressures, plasma osmolality (Posm), and plasma AVP (PAVP) were measured at 0, 12, 16, and 24 h of each WDT. Urine volume was measured at each void throughout testing. Baseline Posm increased from SL to altitude (SL 291.7 +/- 0.8 mosmol/kgH2O, AA 299.6 +/- 2.2 mosmol/kgH2O, PA 302.3 +/- 1.5 mosmol/kgH2O, P < 0.05); however, baseline PAVP measurements were similar. Despite similar Posm values, the maximal PAVP response during the WDT (at 16 h) was greater at altitude than at SL (SL 1.7 +/- 0.5 pg/ml, AA 6.4 +/- 0.7 pg/ml, PA 8.7 +/- 0.9 pg/ml, P < 0.05). In conclusion, hypoxia appeared to alter AVP regulation by raising the osmotic threshold and increasing AVP responsiveness above that threshold.

Improving ultrasound reflectivity and stability of echogenic liposomal dispersions for use as targeted ultrasound contrast agents.

Targeted echogenic liposome dispersions for ultrasonic enhancement of vasoactive and pathological components of endothelium and atherosclerosis have recently been developed. The component lipids required for acoustic and targeting properties include phosphatidylcholine, phosphatidylethanolamine phosphatidylglycerol (PG), and cholesterol (CH), initially in a 60:8:2:30 mol % ratio. Component lipids, lyophilization, sugars, and freezing conditions were varied to optimize acoustic ultrasound reflectivity and acoustic stability. Echogenic liposome dispersions were made by using the dehydration-rehydration process. The lipid concentrations were varied (CH in the range 1 to 40 mol % and PG from 1 to 16 mol %). Variations in type and concentration of sugars were examined. The effect of freezing conditions and re-lyophilization was examined. Ultrasound reflectivity was assessed by using a 20-MHz intravascular ultrasound catheter and computer-assisted videodensitometry. Ultrasound reflectivity was optimized at a CH concentration of 10 mol %; PG concentration variation had essentially no effect on initial values of echogenicity. Optimal acoustic stability was observed with concentrations of 10-15 mol % CH and with a PG concentration greater than 4 mol %. Preparations made with 0.2 M mannitol were more ultrasound reflective than those made with lactose, trehalose, and sucrose. Re-lyophilization and freezing temperatures below -20 degrees C increased ultrasound reflectivity. We optimized the ultrasound properties of echogenic liposomal dispersions, the conditions of which provide some insight into the underlying lipid structures responsible. The preparations developed are now more stable and acoustically reflective than our previous preparations. This advances the development of echogenic lipid dispersions as targeted ultrasound contrast agents for use in general ultrasound as well as cardiovascular imaging.

Detection of melanin-like pigments in the dimorphic fungal pathogen Paracoccidioides brasiliensis in vitro and during infection.

Melanins are implicated in the pathogenesis of several human diseases, including some microbial infections. In this study, we analyzed whether the conidia and the yeasts of the thermally dimorphic fungal pathogen Paracoccidioides brasiliensis produce melanin or melanin-like compounds in vitro and during infection. Growth of P. brasiliensis mycelia on water agar alone produced pigmented conidia, and growth of yeasts in minimal medium with L-3,4-dihydroxyphenylalanine (L-DOPA) produced pigmented cells. Digestion of the pigmented conidia and yeasts with proteolytic enzymes, denaturant, and hot concentrated acid yielded dark particles that were the same size and shape as their propagules. Immunofluorescence analysis demonstrated reactivity of a melanin-binding monoclonal antibody (MAb) with the pigmented conidia, yeasts, and particles. Electron spin resonance spectroscopy identified the yeast-derived particles produced in vitro when P. brasiliensis was grown in L-DOPA medium as a melanin-like compound. Nonreducing polyacrylamide gel electrophoresis of cytoplasmic yeast extract revealed a protein that catalyzed melanin synthesis from L-DOPA. The melanin binding MAb reacted with yeast cells in tissue from mice infected with P. brasiliensis. Finally digestion of infected tissue liberated particles reactive to the melanin binding MAb that had the typical morphology of P. brasiliensis yeasts. These data strongly suggest that P. brasiliensis propagules, both conidia and yeast cells, can produce melanin or melanin-like compounds in vitro and in vivo. Based on what is known about the function of melanin in the virulence of other fungi, this pigment may play a role in the pathogenesis of paracoccidioidomycosis.

Molecular cloning, phylogenetic analysis and three-dimensional modeling of Cu,Zn superoxide dismutase (CnSOD1) from three varieties of Cryptococcus neoformans.

Cryptococcus neoformans (Cn), causal agent of fungal meningoencephalitis, has three varieties with variable host predilection. To explore mechanisms for these pathogenic differences, we have characterized Cu,Zn SOD gene (CnSOD1). A Saccharomyces cerevisiae sod1Delta mutant was complemented with Cn var. grubii yeast expression library. The complementing clone had an ORF of 462 bp and the deduced 154 aa sequence showed 61% identity with S. cerevisiae SOD1 and 53-65% with other eukaryotic SOD1s. Cn var. grubii CnSOD1 cDNA was used to clone corresponding cDNAs from var. neoformans and var. gattii. ORFs from three varieties revealed 20-29% differences in deduced aa (s) with a significant 6% non-synonymous aa substitution between Cn var. grubii and Cn var. gattii. Cosmid library screening and PCR cloning were used to obtain genomic SOD1, which was split by five introns with identical placements and a typical 5' splice junction sequence, GTNNGY. These introns also showed a large nt variation among the three Cn varieties. Phylogenetic analyses revealed CnSOD1 to be in a group distinct from other eukaryotic SOD1s and with a significant divergence of the var. grubii from var. gattii. The CnSOD1 -deduced protein was modeled based on the crystal structure of S. cerevisiae SOD1, which showed an excellent fit. Most of the non-synonymous aa substitutions occurred on the outside of the molecule and these may contribute to differences in antigenicity among the three varieties. Notably, Cn var. neoformans and var. gattii Cu,Zn SOD had three substitutions of glycine (Gly26, Gly92 and Gly123 for Asn26, Ser92 and Ser123) that may contribute to the observed lower thermostability of this enzyme vis-a-vis Cn var. grubii. This is the first nucleotide and structural comparison of a protein-encoding gene from the three Cn varieties, which may provide a framework for future studies on the role of Cu,Zn SOD in Cn pathogenesis.

Production of a species specific monoclonal antibody against a 56-kDa Mycobacterium marinum antigen which is potentially useful in culture identification.

A monoclonal antibody (MoAb) designated 1H11 has been produced against a 56-kDa antigen from Mycobacterium marinum. This MoAb has been shown to be species specific by both enzyme-linked immunosorbent assay and Western blot. The relationship of this 56 kDa to other known mycobacterial antigens is currently under investigation. MoAb 1H11 recognised bacilli taken from cultures by both immunofluorescence and immunoperoxidase staining. This MoAb 1H11 may provide a simple method for rapid identification of M. marinum from cultures of clinical isolates.

Analysis by enzyme-linked immunosorbent assay and immunoblot of the antibody responses of patients infected with Mycobacterium marinum.

The serum antibody responses of a total of 14 patients with active or recently cured Mycobacterium marinum infections were analysed via a combination of enzyme-linked immunosorbent assay (ELISA) and the immunodevelopment of Western blots of M. marinum antigen. Normal human sera and sera from patients with active pulmonary tuberculosis were also analysed as controls. The detectable IgG response of M. marinum patients, as demonstrated by ELISA, was highly variable and did not differ significantly from normal controls. IgA and IgM levels were generally low in the M. marinum patients and were not significantly different from normal controls. Immunodevelopment of Western blots of M. marinum antigen with the sera of patients with M. marinum infections revealed that a number of antigens were recognised. Of particular note was an 18-kDa species that was recognised by 11 out of 14 patients (and by none of the normal controls). The 18-kDa antigen may be a useful serodiagnostic marker in the identification of M. marinum infections.