The Intestinal Tumour Microenvironment.

The tumour microenvironment (TME) of intestinal tumours is highly complex and comprises a network of stromal cells, tumour cells, immune cells and fibroblasts, as well as microorganisms. The tumour location, environmental factors and the tumour cells themselves influence the cells within the TME. Immune cells can destroy tumour cells and are associated with better patient prognosis and response to therapy; however, immune cells are highly plastic and easily influenced to instead promote tumour growth. The interaction between local immune cells and the microbiome can lead to progression or regression of intestinal tumours. In this chapter, we will discuss how tumour development and progression can influence, and be influenced by, the microenvironment surrounding it, focusing on immune and fibroblastic cells, and the intestinal microbiota, particularly in the context of colorectal cancer.

Estimation of genetic parameters for traits associated with reproduction, lactation, and efficiency in sows.

Increased milk production due to high litter size, coupled with low feed intake, results in excessive mobilization of sow body reserves during lactation, which can have detrimental effects on future reproductive performance. A possibility to prevent this is to improve sow lactation performance genetically, along with other traits of interest. The aim of this study was to estimate breed-specific genetic parameters (by parity, between parities, and across parities) for traits associated with lactation and reproduction in Yorkshire and Landrace sows. Performance data were available for 2,107 sows with 1 to 3 parities (3,424 farrowings total). Sow back fat, loin depth and BW at farrowing, sow feed intake (SFI), and body weight loss (BWL) during lactation showed moderate heritabilities (0.21 to 0.37) in both breeds, whereas back fat loss (BFL), loin depth loss (LDL), and litter weight gain (LWG) showed low heritabilities (0.12 to 0.18). Among the efficiency traits, sow lactation efficiency showed extremely low heritability (near zero) in Yorkshire sows but a slightly higher (0.05) estimate in Landrace sows, whereas sow residual feed intake (SRFI) and energy balance traits showed moderate heritabilities in both breeds. Genetic correlations indicated that SFI during lactation had strong negative genetic correlations with body resource mobilization traits (BWL, BFL, and LDL; -0.35 to -0.70), and tissue mobilization traits in turn had strong positive genetic correlations with LWG (+0.24 to +0.54; < 0.05). However, SFI did not have a significant genetic correlation with LWG. These genetic correlations suggest that SFI during lactation is predominantly used for reducing sow body tissue losses, rather than for milk production. Estimates of genetic correlations for the same trait measured in parities 1 and 2 ranged from 0.64 to 0.98, which suggests that first and later parities should be treated as genetically different for some traits. Genetic correlations estimated between traits in parities 1 and 2 indicated that BWF and BWL measured in parity 1 can be used as indicator traits for SFI and SRFI measured in parities 1 and 2. In conclusion, traits associated with lactation in sows have a sizable genetic component and show potential for genetic improvement.

Genetic analysis of reproductive traits and antibody response in a PRRS outbreak herd.

Porcine reproductive and respiratory syndrome (PRRS) is the most economically significant disease impacting pig production in North America, Europe, and Asia, causing reproductive losses such as increased rates of stillbirth and mummified piglets. The objective of this study was to explore the genetic basis of host response to the PRRS virus (PRRSV) in a commercial multiplier sow herd before and after a PRRS outbreak, using antibody response and reproductive traits. Reproductive data comprising number born alive (NBA), number alive at 24 h (NA24), number stillborn (NSB), number born mummified (NBM), proportion born dead (PBD), number born dead (NBD), number weaned (NW), and number of mortalities through weaning (MW) of 5,227 litters from 1,967 purebred Landrace sows were used along with a pedigree comprising 2,995 pigs. The PRRS outbreak date was estimated from rolling averages of farrowing traits and was used to split the data into a pre-PRRS phase and a PRRS phase. All 641 sows in the herd during the outbreak were blood sampled 46 d after the estimated outbreak date and were tested for anti-PRRSV IgG using ELISA (sample-to-positive [S/P] ratio). Genetic parameters of traits were estimated separately for the pre-PRRS and PRRS phase data sets. Sows were genotyped using the PorcineSNP60 BeadChip, and genome-wide association studies (GWAS) were performed using method Bayes B. Heritability estimates for reproductive traits ranged from 0.01 (NBM) to 0.12 (NSB) and from 0.01 (MW) to 0.12 (NBD) for the pre-PRRS and PRRS phases, respectively. S/P ratio had heritability (0.45) and strong genetic correlations with most traits, ranging from -0.72 (NBM) to 0.73 (NBA). In the pre-PRRS phase, regions associated with NSB and PBD explained 1.6% and 3% of the genetic variance, respectively. In the PRRS phase, regions associated with NBD, NSB, and S/P ratio explained 0.8%, 11%, and 50.6% of the genetic variance, respectively. For S/P ratio, 2 regions on SSC 7 (SSC7) separated by 100 Mb explained 40% of the genetic variation, including a region encompassing the major histocompatibility complex, which explained 25% of the genetic variance. These results indicate a significant genomic component associated with PRRSV antibody response and NSB in this data set. Also, the high heritability and genetic correlation estimates for S/P ratio during the PRRS phase suggest that S/P ratio could be used as an indicator of the impact of PRRS on reproductive traits.

Genetic and phenotypic parameters for carcass and meat quality traits in commercial crossbred pigs.

Pork quality and carcass characteristics are now being integrated into swine breeding objectives because of their economic value. Understanding the genetic basis for these traits is necessary for this to be accomplished. The objective of this study was to estimate phenotypic and genetic parameters for carcass and meat quality traits in 2 Canadian swine populations. Data from a genomic selection study aimed at improving meat quality with a mating system involving hybrid Landrace × Large White and Duroc pigs were used to estimate heritabilities and phenotypic and genetic correlations among them. Data on 2,100 commercial crossbred pigs for meat quality and carcass traits were recorded with pedigrees compromising 9,439 animals over 15 generations. Significant fixed effects (company, sex, and slaughter batch), covariates (cold carcass weight and slaughter age), and random additive and common litter effects were fitted in the models. A series of pairwise bivariate analyses were implemented in ASReml to estimate phenotypic and genetic parameters. Heritability estimates (±SE) for carcass traits were moderate to high and ranged from 0.22 ± 0.08 for longissimus dorsi muscle area to 0.63 ± 0.04 for trimmed ham weight, except for firmness, which was low. Heritability estimates (±SE) for meat quality traits varied from 0.10 ± 0.04 to 0.39 ± 0.06 for the Minolta b* of ham quadriceps femoris muscle and shear force, respectively. Generally, most of the genetic correlations were significant (P < 0.05) and ranged from low (0.18 ± 0.07) to high (-0.97 ± 0.35). There were high negative genetic correlations between drip loss with pH and shear force and a positive correlation with cooking loss. Genetic correlations between carcass weight (both hot and cold) with carcass marbling were highly positive. It was concluded that selection for increasing primal and subprimal cut weights with better pork quality may be possible. Furthermore, the use of pH is confirmed as an indicator for pork water-holding capacity and cooking loss. The heritabilities of carcass and pork quality traits indicated that they can be improved using traditional breeding methods and genomic selection, respectively. The estimated genetic parameters for carcass and meat quality traits can be incorporated into the breeding programs that emphasize product quality in these Canadian swine populations.

Estimates of genetic parameters for stayability to consecutive calvings of Canadian Simmentals by random regression models.

Stayability to consecutive calvings was selected as a measure of cow longevity in the Canadian Simmental population. Calving performance data on 188,579 cows and culling information from the Total Herd Reporting System were used to determine whether a cow stayed in a herd for her second and later (up to the eighth) calvings, given that she had calved as 2 yr old. Binary records (n = 1,164,319) were analyzed with animal linear and threshold models including fixed effects of year of birth by season of birth by parity number and age of cow at first calving by parity number and random effects of contemporary group (CG) defined as herd of birth within year by season, animal additive genetic effect, and a cow permanent environmental (PE) effect. All random effects were Legendre polynomial regressions of the same order, defined on the scale from second to the eighth calving. Bayesian methods with Gibbs sampling were used to estimate covariance components and genetic parameters for random effects of models and selected variables on the longitudinal scale. Bayes factors and analyses of mean squared error and correlation between observed and predicted observations indicated that the linear model with regressions of order 3 was most plausible for generating the current data compared with a fixed regression and other random regression (both linear and threshold) models of order up to 4. Estimates of variances for all random effects from the best fitting model changed with the calving number. Estimates of heritability decreased in time: from 0.35 (SD = 0.006) for stayability to second calving to 0.13 (SD = 0.004) for stayability to the eighth calving. Variance due to PE effect constituted the largest part of the total variance of stayability for all longitudinal points followed by genetic and CG components. Genetic effects of stayability to different calvings were relatively highly correlated, from 0.62 (SD = 0.011) to 0.99 (SD = 0.001), and correlation decreased with the time span between calvings. Correlations for PE and CG effects showed similar trends. Animal genetic effect seemed to be less variable on the longitudinal scale compared with other random effects of the model. The first 2 principal components explained from 95% (PE effects) to 99% (genetic effect) of the total variance. The overall level of genetic stayability curve correlated well (from 0.87 to 0.99, with SD < 0.006) with genetic stayability to different calvings and therefore could be used as a single criterion in selection for stayability.

Myocardial blood flow in patients with low-flow, low-gradient aortic stenosis: differences between true and pseudo-severe aortic stenosis. Results from the multicentre TOPAS (Truly or Pseudo-Severe Aortic Stenosis) study.

BACKGROUND: Impairment of myocardial flow reserve (MFR) in aortic stenosis (AS) with normal left ventricular function relates to the haemodynamic severity. OBJECTIVES: To investigate whether myocardial blood flow (MBF) and MFR differ in low-flow, low-gradient AS depending on whether there is underlying true-severe AS (TSAS) or pseudo-severe AS (PSAS). METHODS: In 36 patients with low-flow, low-gradient AS, dynamic [13N]ammonia PET perfusion imaging was performed at rest (n = 36) and during dipyridamole stress (n = 20) to quantify MBF and MFR. Dobutamine echocardiography was used to classify patients as TSAS (n = 18) or PSAS (n = 18) based on the indexed projected effective orifice area (EOA) at a normal flow rate of 250 ml/s (EOAI(proj )0.55 cm(2)/m(2)). RESULTS: Compared with healthy controls (n = 14), patients with low-flow, low-gradient AS had higher resting mean (SD) MBF (0.83 (0.21) vs 0.69 (0.09) ml/min/g, p = 0.001), reduced hyperaemic MBF (1.16 (0.31) vs 2.71 (0.50) ml/min/g, p<0.001) and impaired MFR (1.44 (0.44) vs 4.00 (0.91), p<0.001). Resting MBF and MFR correlated with indices of AS severity in low-flow, low-gradient AS with the strongest relationship observed for EOAI(proj) (r(s) = -0.50, p = 0.002 and r(s) = 0.61, p = 0.004, respectively). Compared with PSAS, TSAS had a trend to a higher resting MBF (0.90 (0.19) vs 0.77 (0.21) ml/min/g, p = 0.06), similar hyperaemic MBF (1.16 (0.31) vs 1.17 (0.32) ml/min/g, p = NS), but a significantly smaller MFR (1.19 (0.26) vs 1.76 (0.41), p = 0.003). An MFR <1.8 had an accuracy of 85% for distinguishing TSAS from PSAS. CONCLUSIONS: Low-flow, low-gradient AS is characterised by higher resting MBF and reduced MFR that relates to the AS severity. The degree of MFR impairment differs between TSAS and PSAS and may be of value for distinguishing these entities.

Weaning, yearling, and preharvest ultrasound measures of fat and muscle area in steers, bulls, and heifers.

Longissimus muscle area and fat thickness were measured following weaning, at yearling, and prior to harvest using real-time ultrasound, and corresponding carcass measurements were recorded 3 to 7 d following the preharvest scan in composite steers (n = 116, 447 +/- 19 d), bulls (n = 224, 521 +/- 11 d), and heifers (n = 257,532 +/- 12 d). Although fat deposition was limited in bulls and heifers from weaning to yearling, coefficients of variation ranged from 8.46 to 13.46% for muscle area, and from 27.55 to 38.95% for fat thickness, indicating that significant phenotypic variance exists across genders. Residual correlations, adjusted for the effects of year of birth, gender, and age at measurement, were high and ranged from 0.79 to 0.87 among ultrasound and carcass measures of muscle area. Residual correlations among ultrasound and carcass measures of fat thickness were also high, ranging from 0.64 to 0.86. Weaning and/or yearling ultrasound muscle area yielded similarly accurate predictions of carcass muscle area. Yearling ultrasound fat thickness accounted for 13% more of the observed variance in carcass fat thickness than the weaning ultrasound measure in single-trait prediction models. When both weaning and yearling ultrasound measures were used to predict carcass fat thickness, partial R2 values were 0.15 and 0.61 for weaning and yearling ultrasound fat thickness, respectively. The difference between predicted and carcass measures with respect to muscle area (fat thickness) was less than 6.45 cm2 (2.5 mm) for 80.2 to 88.9% (90.3 to 95%) of animals. Preharvest ultrasound measures yielded standard errors of prediction of less than 4.95 cm2 for muscle area and 1.51 mm or less for fat thickness. These results indicate that ultrasound measures taken between weaning and yearling provide accurate predictors of corresponding carcass traits in steers, bulls, and heifers.

Genetic evaluation of carcass yield using ultrasound measures on young replacement beef cattle.

Live weight and ultrasound measures of fat thickness and longissimus muscle area were available on 404 yearling bulls and 514 heifers, and carcass measures of weight, longissimus muscle area, and fat thickness were available on 235 steers. Breeding values were initially estimated for carcass weight, longissimus muscle area, and fat thickness using only steer carcass data. Breeding values were also estimated for weight and ultrasound muscle area and fat thickness using live animal data from bulls and heifers, with traits considered sex-specific. The combination of live animal and carcass data were also used to estimate breeding values in a full animal model. Breeding values from the carcass model were less accurate and distributed more closely around zero than those from the live data model, which could at least partially be explained by differences in relative amounts of data and in phenotypic mean and heritability. Adding live animal data to evaluation models increased the average accuracy of carcass trait breeding values 91, 75, and 51% for carcass weight, longissimus muscle area, and fat thickness, respectively. Rank correlations between breeding values estimated with carcass vs live animal data were low to moderate, ranging from 0.16 to 0.43. Significant rank changes were noted when breeding values for similar traits were estimated exclusively with live animal vs carcass data. Carcass trait breeding values estimated with both live animal and carcass data were most accurate, and rank correlations reflected the relative contribution of carcass data and their live animal indicators. The addition of live animal data to genetic evaluation of carcass traits resulted in the most significant carcass trait breeding value accuracy increases for young replacements that had not yet produced progeny with carcass data.

Tumor-specific Tc1, but not Tc2, cells deliver protective antitumor immunity.

We investigated whether secretion of multiple cytokines by CD8+ T cells is associated with improved protection against tumor challenge. We show that antitumor immunity induced by immunization with dendritic cells and a MHC class I-binding tumor peptide are dependent on secretion of IFN-gamma but not IL-4 or IL-5 by host cells. To further address the role of IL-4 and IL-5 in antitumor immunity, tumor-specific TCR-transgenic CD8+ T cells were activated in vitro to generate cytotoxic T (Tc) 1 cells that secrete high IFN-gamma and no IL-4 or IL-5 or Tc2 cells that secrete IL-4, IL-5, and some IFN-gamma. Both cell types killed target cells in vitro. Tc1 and Tc2 cells were adoptively transferred into syngeneic hosts, and their ability to protect against tumor challenge was compared. Tc1 cells were able to significantly delay tumor growth, whereas Tc2 cells or Tc2 cells from IFN-gamma(-/-) donors had no effect. This was due to neither the inability of Tc2 cells to survive in vivo or to migrate to the tumor site nor their inability to secrete IL-4 and/or IL-5 in the presence of limiting amounts of anti-CD3. However, IFN-gamma secretion by Tc2 cells was triggered inefficiently by restimulation with Ag compared with anti-CD3. We conclude that the ability to secrete "type 2" cytokines, and cytotoxic ability, have a limited role in antitumor immune responses mediated by CD8+ T cells, whereas the capacity to secrete high amounts of IFN-gamma remains the most critical antitumor effector mechanism in vivo.

Effects of rooting and tree growth of selected woodland species on cap integrity in a mineral capped landfill site.

The above and below ground growth of three tree species (Alnus glutinosa, Pinus nigra var. maritima and Acer pseudoplatanus) was studied on a containment landfill site at Waterford, Hertfordshire, UK. Tree root architecture was studied using soil inspection pits excavated next to 12 trees of each species and mapped in detail. Tree height was related to soil thickness over the compacted mineral cap. No roots entered the cap where soil thickness was 1.3 m, but a few roots, especially of alder, were observed within it when the soil cover was 1.0 m or less. Micromorphological analysis of undisturbed samples of the mineral cap suggested that roots exploited weaknesses in the cap rather than actively causing penetration into it. Alder roots were more tolerant of anaerobic conditions within the cap than the other species examined. The results confirm that mineral caps should be covered by 1.5 m of soil or soil-forming material if tree establishment is intended over a restored landfill site, unless protected by other parts of a composite capping system.

Gene microarrays reveal extensive differential gene expression in both CD4(+) and CD8(+) type 1 and type 2 T cells.

An important subdivision of effector T cells can be made based on patterns of cytokine production and functional programs. Type 1 T cells produce IFN-gamma and protect against viral pathogens, whereas type 2 cells produce cytokines such as IL-4 and IL-5 and protect against large extracellular parasites. Both CD4(+) and CD8(+) T cells can be polarized into type 1 or type 2 cytokine-secreting cells, suggesting that both populations play a regulatory role in immune responses. In this study, we used high-density oligonucleotide arrays to produce a comprehensive picture of gene expression in murine CD4(+) Th1 and Th2 cells, as well as CD8(+) type 1 and type 2 T cells. Polarized type 1 and 2 cells transcribed mRNA for an unexpectedly large number of genes, most of which were expressed in a similar fashion between type 1 and type 2 cells. However, >100 differentially expressed genes were identified for both the CD4(+) and CD8(+) type 1 and 2 subsets, many of which have not been associated with T cell polarization. These genes included cytokines, transcription factors, molecules involved in cell migration, as well as genes with unknown function. The program for type 1 or type 2 polarization was similar for CD4(+) and CD8(+) cells, since gene expression patterns were roughly the same. The expression of select genes was confirmed using real-time PCR. The identification of genes associated with T cell polarization may give important insights into functional and phenotypic differences between effector T cell subsets and their role in normal responses and inflammatory disease.

Genetic parameters for ultrasound and carcass measures of yield and quality among replacement and slaughter beef cattle.

Real time ultrasound (RTU) measures of longissimus muscle area and fat depth were taken at 12 and 14 mo of age on composite bulls (n = 404) and heifers (n = 514). Carcass longissimus muscle area and fat depth, hot carcass weight, estimated percentage lean yield, marbling score, Warner-Bratzler shear force, and 7-rib dissectable seam fat and lean percentages were measured on steers (n = 235). Additive genetic variances for longissimus muscle area were 76 and 77% larger in bulls at 12 and 14 mo than the corresponding estimates for heifers. Heritability estimates for longissimus muscle area were 0.61 and 0.52 in bulls and 0.49 and 0.47 in heifers at 12 and 14 mo, respectively. The genetic correlations of longissimus muscle area of bulls vs heifers were 0.61 and 0.84 at 12 and 14 mo, respectively. Genetic correlations of longissimus muscle area measured in steer carcasses were 0.71 and 0.67 with the longissimus muscle areas in bulls and heifers at 12 mo and 0.73 and 0.79 at 14 mo. Heritability estimates for fat depth were 0.50 and 0.35 in bulls and 0.44 and 0.49 in heifers at 12 and 14 mo, respectively. The genetic correlation of fat depth in bulls vs heifers at 12 mo was 0.65 and was 0.49 at 14 mo. Genetic correlations of fat depth measured in bulls at 12 and 14 mo with fat depth measured in steers at slaughter were 0.23 and 0.21, and the corresponding correlations of between heifers and steers were 0.66 and 0.86, respectively. Live weights at 12 and 14 mo were genetically equivalent (r(g) = 0.98). Genetic correlations between live weights of bulls and heifers with hot carcass weight of the steers were also high (r(g) > 0.80). Longissimus muscle area measured using RTU was positively correlated with carcass measures of longissimus muscle area, estimated percentage lean yield, and percentage lean in a 7-rib section from steers. Measures of backfat obtained using RTU were positively correlated with fat depth and dissectable seam fat from the 7-rib section of steer carcasses. Genetic correlations between measures of backfat obtained using RTU and marbling were negative but low. These results indicate that longissimus muscle area and backfat may be under sufficiently different genetic control in bulls vs heifers to warrant being treated as separate traits in genetic evaluation models. Further, traits measured using RTU in potential replacement bulls and heifers at 12 and 14 mo of age may be considered different from the corresponding carcass traits of steers.

Dendritic cell elimination as an assay of cytotoxic T lymphocyte activity in vivo.

We show in this paper that the survival of antigen-loaded dendritic cells in vivo may be used as a sensitive readout of CTL activity. We have previously shown that dendritic cells labeled with the fluorescent dye CFSE and injected sub-cutaneously into mice migrate spontaneously to the draining lymph node where they persist for several days. In the presence of effector CTL responses, dendritic cells loaded with specific antigen rapidly disappear from the draining lymph node. In this paper we extend the above observations and set up a simple and sensitive method to reveal CTL activity in individual mice in vivo. Dendritic cells were labeled with two different fluorochromes, loaded with antigen or left untreated, and mixed together before injection into mice. We show that only the dendritic cells loaded with specific antigen were cleared from the draining lymph node, while dendritic cells not loaded with antigen remained unaffected. Cytotoxic responses generated by immunization with peptide-loaded dendritic cells, or by infection with influenza virus, could be revealed using this method. Comparison of the differential survival of dendritic cells populations mixed together also allowed us to accurately evaluate the disappearance of dendritic cells, irrespective of variability in the injection site and other parameters. Given the ability of dendritic cells to efficiently take up and present complex antigens, nucleic acids and apoptotic bodies, this method may also allow the evaluation of cytotoxic activity against antigens that are not characterized in terms of peptide epitopes.

Comparison of models for genetic evaluation of scrotal circumference in crossbred bulls.

The aim of this study was to evaluate the effect of including concomitant body weight and(or) a random dam effect in genetic evaluation models on variance component estimates and standard error of prediction for scrotal circumference (SC) at 6, 8, 10, and 12 mo. Variance components and average standard errors of prediction were compared under models differing in either the number of related traits (M11 [SC], M12 [SC and BW]) or an uncorrelated random dam effect (M21 [SC], M22 [SC and BW]) using records on 1,547 bull calves. In a single-trait model (M11), estimates of direct heritabilities (h2a) for SC were .45, .49, .57, and .66 at 6, 8, 10, and 12 mo, respectively. In a two-trait model (M12), h2a were similar to those in M11 model. In M21, h2a for SC were .37, .42, .54, and .65, whereas the proportions of phenotypic variance due to dams (d2) were .12, .11, .04, and .02 at 6, 8, 10, and 12 mo, respectively. Similarly, in M22, h2a for SC were .36, .44, .56, and .65 and d2 were .13, .10, .02, and .02. Standard errors of prediction for SC EBV from M22 were reduced by 2.86, 1.21, 3.02, and 1.99% relative to M21 and by 6.45, 2.70, 2.72, and 1.21% relative to M11 at 6, 8, 10, and 12 mo, respectively. Standard errors of prediction for SC EBV from M12 were reduced by .06, .73, 1.56, and .87% relative to M11 at 6, 8, 10, and 12 mo, respectively. The importance of the dam effect decreased with age for both SC and BW. These results demonstrate that a two-trait (SC and BW) animal model would result in more accurate evaluations of yearling SC EBV in beef cattle than a single-trait model.

Survival of Listeria monocytogenes in sea water and effect of exposure on thermal resistance.

Survival, recoverability and sublethal injury of two strains of Listeria monocytogenes, Scott A and an environmental strain KM, on exposure to sea water at 12.8 or 20.8 degrees C was determined using in situ diffusion chambers. Plate counts were used to assess recoverability and injury while 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) reduction was used to determine respiratory activity. T90 values (times for 10-fold decreases in numbers of recoverable cells) on non-selective medium (trypticase soya agar with 0.6% yeast extract) at 12.8 and 20.8 degrees C were 61.7 and 69.2 h for L. monocytogenes Scott A, and 103.0 and 67.0 h for L. monocytogenes KM, respectively. On selective medium (Oxford agar), T90 values at 12.8 and 20.8 degrees C were 60.6 and 56.9 h for L. monocytogenes Scott A, and 83.0 and 65.9 h for L. monocytogenes KM, respectively. With Scott A, the percentage of sublethally injured cells at 12.8 and 20.8 degrees C was 1.7 and 17.7%, respectively, while for KM the values were 19.0 and 1.6%, respectively. The fraction of cells reducing CTC but which were not recoverable on plating progressively increased on exposure to sea water. Listeria monocytogenes KM challenged at 58 degrees C showed an apparent increase in heat resistance after exposure to sea water at 20.8 degrees C for 7 d (D58 = 2.64 min) compared with before exposure (D58 = 1.24). This increase in thermal resistance was not apparent at temperatures greater than 63 degrees C, and analysis of the best-fit regression lines fitted to the thermal data obtained from the two cell populations indicated that their thermal resistance was not significantly different (P > 0.05) over the temperature range tested (58-62 degrees C).

Thermal death times of Hafnia alvei cells in a model suspension and in artificially contaminated hot-smoked kahawai (Arripis trutta).

In New Zealand the product most frequently implicated in cases of scombroid poisoning is hot-smoked kahawai (Arripis trutta). Using a Hafnia alvei strain, previously isolated from a portion of hot-smoked kahawai with a histamine level of 1,659.4 mg/kg, thermal death trials were carried out in a model suspension (0.1% peptone) at 54, 55, 56, 57, and 58 degrees C. From the linear regression line (R2 = 0.98) fitted to observed D values plotted against temperature, calculated D values for 54, 55, 56, 57, and 58 degrees C were estimated to be 0.63, 0.36, 0.20, 0.11, and 0.06 min, respectively, giving a z value of 4.14 degrees C. Thermal death trials were also carried out for H. alvei associated with hot-smoked kahawai at 54, 55, 55.5, 56, and 57 degrees C. From the linear regression line (R2 = 0.93) fitted to the data, calculated D values for 54, 55, 56, and 57 degrees C were estimated to be 1.42, 0.74, 0.38, and 0.20 min, respectively, giving a z value of 3.57 degrees C. Results indicate that hot smoking has the potential to eliminate H. alvei from seafood products.

The Family Project Approach.

The Family Project Approach (FPA) has been developed as an alternative to the placement of conduct-disordered adolescents into care. Since 1987, the target group of this project has been extended to include maltreated children and to parents who are in despair because of persistent child behaviour problems and other stressors. This article gives an overview of this home-based approach and the most important research results. One main starting point for FPA is that parents should be supported so that they can fulfil their child-rearing responsibilities. In families where the children are neglected or maltreated and in families where the parents feel desperate and powerless, the fundamental attitude of the FPA worker plays a decisive role. Process-outcome research corroborates the importance of encouraging and validating the parents' positive qualities and expertise.

Opening the black box: the relationship between neural networks and linear discriminant functions.

Over the last ten years feed-forward neural networks have become a popular tool for statistical decision making. During this time, they have been applied in many fields, including cytological classification. Neural networks are often treated as a black box, whose inner workings are concealed from the researcher. This is unfortunate, since the inner workings of a neural network can be understood in a manner similar to that of a linear discriminant function, which is the standard tool that researchers use for decision making. This paper discusses feed-forward neural networks and some methods to improve their performance for classification problems. Their relationship to discriminant functions will be examined for a simple two-dimensional classification problem.

Detection of malignancy associated changes in cervical cell nuclei using feed-forward neural networks.

Normal cells in the presence of a precancerous lesion undergo subtle changes of their DNA distribution when observed by visible microscopy. These changes have been termed Malignancy Associated Changes (MACs). Using statistical models such as neural networks and discriminant functions it is possible to design classifiers that can separate these objects from truly normal cells. The correct classification rate using feed-forward neural networks is compared to linear discriminant analysis when applied to detecting MACs. Classifiers were designed using 53 nuclear features calculated from images for each of 25,360 normal appearing cells taken from 344 slides diagnosed as normal or containing severe dysplasia. A linear discriminant function achieved a correct classification rate of 61.6% on the test data while neural networks scored as high as 72.5% on a cell-by-cell basis. The cell classifiers were applied to a library of 93,494 cells from 395 slides, and the results were jackknifed using a single slide feature. The discriminant function achieved a correct classification rate of 67.6% while the neural networks managed as high as 76.2%.

Therapist-parent interaction patterns in home-based treatments: exploring family therapy process.

At the current stage of development in the family therapy field, exploratory, small-scale process studies are necessary in order to understand through what interpersonal processes child and family change occur. The goal of this article is to show how relevant therapist-parent interactions within family-based approaches can be explored and linked to the reduction in childrearing and behavior problems. Sequential analyses on 13 treatments using the Family Project Approach revealed that, within the most successful treatments, therapist and parent interact in a collaborative way in the phase of Direct Influence. In the beginning of this treatment phase, the therapist must activate the parents to tackle the problems actively. Further explorations indicated that a collaborative interaction pattern between therapist and mother during the first three sessions of therapy contributes to a better outcome.