Growth and feed efficiency of Nordic Red Dairy Cattle, Holstein, and their F1 crossbreeds when limiting feed energy concentration in prepubertal heifers.

Milk production and overall dairy farm economics depend on rearing dairy heifers. This study investigated the presence of a genotype by environment interaction in Holstein (HOL), Nordic Red Dairy Cattle (RDC), and their F1 crossbreeds (HOLxRDC) when provided different feed rations. The aim of our study was to assess how different energy concentrations in feed rations affect growth, body condition scores, feed intake, and feed efficiency in the 3 groups during the prepubertal period. The 3 breed groups were randomly allocated to receive either a standard or a low energy feed ration. HOL heifers exhibited reduced growth and a lower body condition score when they were fed the low energy feed ration. In contrast, the RDC heifers demonstrated similar growth rates with the different feed rations and maintained similar body condition scores irrespective of feed energy concentration. HOLxRDC crossbred heifers performed as an intermediate between the HOL and RDC groups. There were significant differences in dry matter intake and energy intake in the HOL and HOLxRDC groups depending on feed ration treatment. The RDC heifers had similar feed intake irrespective of treatment. There were no significant differences in the feed conversion ratio between breeds and feed treatments. These results indicate the presence of a genotype by environment interaction in prepubertal HOL and RDC heifers in response to differences in feed ration treatment. Due to the influence of prepubertal growth on future milk production, reproduction, and health status, it is important to be aware of breed-specific requirements during the prepubertal period, particularly in mixed-breed and crossbred groups, to optimize growth rates and production potential.

Physical and barrier changes in gastrointestinal mucus induced by the permeation enhancer sodium 8-[(2-hydroxybenzoyl)amino]octanoate (SNAC).

Drug delivery systems (DDS) for oral delivery of peptide drugs contain excipients that facilitate and enhance absorption. However, little knowledge exists on how DDS excipients such as permeation enhancers interact with the gastrointestinal mucus barrier. This study aimed to investigate interactions of the permeation enhancer sodium 8-[(2-hydroxybenzoyl)amino]octanoate (SNAC) with ex vivo porcine intestinal mucus (PIM), ex vivo porcine gastric mucus (PGM), as well as with in vitro biosimilar mucus (BM) by profiling their physical and barrier properties upon exposure to SNAC. Bulk mucus permeability studies using the peptides cyclosporine A and vancomycin, ovalbumin as a model protein, as well as fluorescein-isothiocyanate dextrans (FDs) of different molecular weights and different surface charges were conducted in parallel to mucus retention force studies using a texture analyzer, rheological studies, cryo-scanning electron microscopy (cryo-SEM), and single particle tracking of fluorescence-labelled nanoparticles to investigate the effects of the SNAC-mucus interaction. The exposure of SNAC to PIM increased the mucus retention force, storage modulus, viscosity, increased nanoparticle confinement within PIM as well as decreased the permeation of cyclosporine A and ovalbumin through PIM. Surprisingly, the viscosity of PGM and the permeation of cyclosporine A and ovalbumin through PGM was unaffected by the presence of SNAC, thus the effect of SNAC depended on the regional site that mucus was collected from. In the absence of SNAC, the permeation of different molecular weight and differently charged FDs through PIM was comparable to that through BM. However, while bulk permeation of neither of the FDs through PIM was affected by SNAC, the presence of SNAC decreased the permeation of FD4 and increased the permeation of FD150 kDa through BM. Additionally, and in contrast to observations in PIM, nanoparticle confinement within BM remained unaffected by the presence of SNAC. In conclusion, the present study showed that SNAC altered the physical and barrier properties of PIM, but not of PGM. The effects of SNAC in PIM were not observed in the BM in vitro model. Altogether, the study highlights the need for further understanding how permeation enhancers influence the mucus barrier and illustrates that the selected mucus model for such studies should be chosen with care.

Investigating the effect of graphene oxide in chitosan/alginate-based foams on the release and antifungal activity of clotrimazole in vitro.

Polyelectrolyte complexes (PECs) have been used as the matrix of solid foams for drug delivery. This study aimed at investigating the effect of graphene oxide (GO) and the composition of excipients in chitosan/alginate-based buccal foams on the clotrimazole release and antifungal activities. The investigation has been focused on the interactions of the drug with excipients in the foams, and the changes of ionization degree upon exposure to various media are discussed. The solid foams were prepared by mixing the excipients and clotrimazole via probe sonication, followed by a freeze-drying method. The pH values of the formulations were measured during the foam preparation process to estimate the ionization degree of clotrimazole and the other excipients. The foam matrix was the PECs between the cationic chitosan and anionic alginate. The mechanical strength of clotrimazole-loaded foams was lower than that of drug-free foams due to the positively charged clotrimazole interacting with the anionic alginate and interfering the PECs between chitosan and alginate. Addition of GO in the clotrimazole-loaded matrix made the foams mechanically stronger and contributed to a faster release of clotrimazole from the buccal foams by disrupting the electrostatic interactions between alginate and clotrimazole. However, addition of 1 wt% GO in the formulations didn't affect the antifungal activity of clotrimazole-loaded foams significantly. A lower amount GO in the formulation may be required for enhancing the antifungal effect, which should be further investigated in future.

Competitive gene flow does not necessarily maximize the genetic gain of genomic breeding programs in the presence of genotype-by-environment interaction.

National and international across-population selection is often recommended and fairly common in the current breeding practice of dairy cattle, with the primary aims to increase genetic gain and genetic variability. The aim of this study was to test the hypothesis that the strategy of truncation selection of sires across populations [i.e., competitive gene flow strategy (CGF)] may not necessarily maximize genetic gain in the long term in the presence of genotype-by-environment interaction (G×E). Two alternative strategies used to be compared with CGF were forced gene flow (FGF) strategies, with 10 or 50% of domestic dams forced to be mated with foreign sires (FGF10%, FGF50%). Two equal-size populations (Ndams = 1,000) that were selected for the same breeding goal trait (h2 = 0.3) under G×E correlation (rg) of either 0.9 or 0.8 were simulated to test these 3 different strategies. Each population first experienced either 5 or 20 differentiation generations (Gd), then 15 migration generations. Discrete generations were simulated for simplicity. Each population performed a within-population conventional breeding program during differentiation generations and the 3 across-population sire selection strategies based on joint genomic prediction during migration generations. The 4 Gd_rg combinations defined 4 different levels of differentiation degree between the 2 populations at the start of migration. The true rate of inbreeding over the last 10 migration generations in each scenario was constrained at 0.01 to provide a fair basis for comparison of genetic gain across scenarios. Results showed that CGF maximized the genetic gain after 15 migration generations in 5_0.9 combination only, the case of the lowest differentiation degree, with a superiority of 0.4% (0.04 genetic SD units) over the suboptimal strategy. While in 5_0.8, 20_0.9, and 20_0.8 combinations, 2 FGF strategies had a superiority in genetic gain of 2.3 to 12.5% (0.21-1.07 genetic SD units) over CGF after 15 migration generations, especially FGF50%. The superiority of FGF strategies over CGF was that they alleviated inbreeding, introduced new genetic variance in the early migration period, and improved accuracy in the entire migration period. Therefore, we concluded that CGF does not necessarily maximize the genetic gain of across-population genomic breeding programs given moderate G×E. The across-population selection strategy remains to be optimized to maximize genetic gain.

Short communication: Heterosis and breed effects for milk production and udder health traits in crosses between Danish Holstein, Danish Red, and Danish Jersey.

During the last decade, the use of systematic crossbreeding in dairy cattle herds has increased in several countries of the world. The aim of this study was to estimate the effect of breed proportion and heterosis on milk production traits and udder health traits in dairy cattle. The study was based on records on milk yield (MY), protein yield (PY), fat yield (FY), somatic cell score (SCS), and mastitis (MAST) from 73,695 first-lactation dairy cows in 130 Danish herds applying systematic crossbreeding programs. Around 45% of the cows were crosses between Danish Holstein (DH), Danish Red (DR), or Danish Jersey (DJ), and the remaining were purebred DH, DR, or DJ. The statistical model included the fixed effects of herd-year, calving month, and calving age and an effect representing the lactation status of the cow. In addition, the model included a regression on calving interval from first to second lactation, a regression on the proportion of DH, DR, and DJ genes, and a regression on the degree of heterozygosity between DH and DR, DH and DJ, and DR and DJ. Random effects were the genetic effect of the cow and a residual. The effect of breed proportions was estimated relatively to DH. For MY, a pure DR yielded 461 kg milk less than DH, whereas a pure DJ yielded 2,259 kg milk less than a pure DH. Compared with DH, PY was 41.7 kg less for DJ, whereas PY for DR was 4.0 kg less than for DH. For FY, a DR yielded 10.6 kg less than DH, whereas there was no significant effect of breed proportion between DJ and DH. A DR cow had lower SCS (0.13) than DH, whereas DJ had higher SCS (0.14) than DH. There was no significant effect of breed proportion on MAST between the 3 breeds. Heterosis was significant in all combinations of breeds for MY, FY, and PY. Heterosis for crosses between DH and DR was 257 kg (3.2%), 11.9 kg (3.2%), and 8.9 kg (3.2%) for MY, PY, and FY, respectively. Corresponding figures for crosses between DH and DJ were 314 kg (4.4%), 14.3 kg (4.4%), and 10.4 kg (4.0%), whereas heterosis between DR and DJ was 462 kg (6.7%), 19.6 kg (6.7%), and 13.9 kg (5.4%) for MY, PY, and FY, respectively. Heterosis was only significant for SCS in the crosses between DH and DR. Heterosis effects for MAST were nonsignificant for all the crosses. The results obtained in this study demonstrate that in first lactation cows, there is a positive effect of heterosis on milk production traits, but limited effect on udder health traits.

Graphene oxide as a functional excipient in buccal films for delivery of clotrimazole: Effect of molecular interactions on drug release and antifungal activity in vitro.

Graphene oxide (GO) is an amphiphilic, high surface area material with great potential as a functional excipient in drug delivery. The present study aimed at incorporating GO in buccal polyelectrolyte films for delivery of antifungal drugs and investigating the effect of GO on the film properties and drug release profiles, as well as antifungal activities. Mucoadhesive excipients chitosan and alginate were selected to form polyelectrolyte films with the antifungal drug clotrimazole. The buccal formulations were prepared by mixing the excipients and clotrimazole via probe sonication, followed by film casting and drying. Inclusion of GO in the formulations increased clotrimazole release from the films in vitro (pH 6.8), possibly due to GO altering the electrostatic interactions between chitosan and alginate. An increase of in vitro activity against Candida albicans was observed when 0.04 wt% GO was added in the formulation containing clotrimazole. However, when the GO amount increased to 0.09 wt%, the films had similar antifungal ability to the films with 0.04 wt% GO, suggesting that the electrostatic and hydrophobic interactions between GO and clotrimazole also affects the antifungal effect of clotrimazole. In summary, GO has a great potential as a functional excipient for delivery of antifungal drugs.

Genetic analysis of vibriosis and viral nervous necrosis resistance in Atlantic cod (Gadus morhua L.) using a cure model.

The aim of this study was to investigate whether observed time-until-death of Atlantic cod (Gadus morhua L.) juveniles in separate challenge tests with Vibrio anguillarum (causes vibriosis) and nodavirus [causes viral nervous necrosis (VNN)] are due to differences in susceptibility (whether at risk or not) or increased endurance (individual hazard, given that the animal is susceptible) using a cure mixture (CURE) model with Gibbs sampling. Observed time-until-death, prepared as sequential binary records, were analyzed with the CURE model and results were compared with cross-sectional threshold (SIMPLE) and an ordinary longitudinal survival score (NAÏVE) model (i.e., assuming that all animals are susceptible). Overall mortality at the end of the test was 86 and 71% for vibriosis and VNN, respectively. But the CURE model estimated 92 and 82% of the population to be susceptible to vibriosis and VNN, respectively. Hence, a substantial fraction among the survivors were considered to be susceptible but with high endurance. The underlying heritability of susceptibility was moderate for vibriosis (0.33) and extremely high for VNN (0.91), somewhat greater compared with classical SIMPLE model (0.19 and 0.76 for vibriosis and VNN, respectively), analyzing end survival as a cross-sectional binary trait. Estimates of the underlying heritability were low for single test-day scores of both endurance (0.02 and 0.15 for vibriosis and VNN, respectively) in the CURE model and for the NAÏVE model (0.02 and 0.18 for vibriosis and VNN, respectively). Based on the CURE model, the genetic correlation between susceptibility and endurance was low to moderately positive and significantly different from unity (P < 0.01) for both vibriosis (0.13) and VNN (0.47). Estimated breeding values from the SIMPLE and NAÏVE models showed moderate to high correlations (0.41 to 0.96) with EBV for susceptibility and endurance in the CURE model. The analyses indicate that susceptibility and endurance are apparently distinct genetic traits. Still, the genetic variation estimated in the SIMPLE and NAÏVE models seems to a large extent to be controlled by susceptibility and an efficient genetic selection for reduced susceptibility to vibriosis and VNN is therefore likely feasible even when using classical (noncure) models. Earlier termination of the challenge test or back truncation of survival data is not recommended as this likely shifts the focus of selection towards endurance rather than susceptibility.

In vitro penetration properties of solid lipid nanoparticles in intact and barrier-impaired skin.

Treatment of skin diseases implies application of a drug to skin with an impaired epidermal barrier, which is likely to affect the penetration profile of the drug substance as well as the carrier into the skin. To elucidate this, the effect of skin barrier damage on the penetration profile of a corticosteroid applied in solid lipid nanoparticles (SLN) composed of different lipids, varying in polarity, was studied. The studies were carried out in vitro using impaired and intact porcine ear skin, and the SLN were compared with a conventional ointment. It was shown that a significantly higher amount of corticosteroid remained in the skin, intact as well as barrier impaired, when SLN was used as a vehicle. In general, the penetration profile of the drug substance into the skin was affected by the type of lipid used in the formulation and related to lipid polarity and drug substance solubility. When formulated in SLN and applied to intact skin, the permeation of the drug substance across the skin was significantly reduced, as compared to the ointment. Altogether, in both barrier-impaired and intact skin, a higher amount of drug substance remained in the skin during application of SLN for 6, 16, and 24h, as compared to the ointment. These results emphasize the applicability of SLN to create a drug reservoir in skin, with the drug localized distinctively in the stratum corneum.

In vivo clearance and metabolism of recombinant activated factor VII (rFVIIa) and its complexes with plasma protease inhibitors in the liver.

INTRODUCTION: Recombinant activated factor VII (rFVIIa, NovoSeven®) is injected intravenously for the treatment of haemophilia patients with inhibitory antibodies. In plasma, rFVIIa forms complexes with protease inhibitors, primarily antithrombin III (ATIII). The liver is believed to be involved in clearance of rFVIIa, however, it is not known whether the liver is also involved for the clearance of the rFVIIa-ATIII complex. In this study, we explored the fate of intravenously injected rFVIIa from plasma to the hepatic lysosomes. MATERIALS AND METHODS: A novel method using magnetic chromatography was used to isolate catabolic organelle (CO) fractions from mouse liver following injection of superparamagnetic dextran (SPD)-coated iron oxide particles and rFVIIa. The effect of co-circulating SPD particles on rFVIIa pharmacokinetic (PK) parameters was evaluated by ELISA. Cryo-immuno transmission electron microscopy (TEM) was used to study hepatic distribution of SPD particles and rFVIIa. The isolated hepatic CO fractions were characterized using Western Blotting (WB). RESULTS: Cryo-immuno TEM of the liver confirmed hepatic co-localisation of SPD particles and rFVIIa in identical endosomes and lysosomes of both hepatocytes and Kupffer cells. SPD particles did not affect the PK parameters of rFVIIa. WB analysis of plasma and CO fractions detected rFVIIa as the full-length protein and also in high molecular weight (HMW) complexes with ATIII and α-2 macroglobulin (α-2M). CONCLUSIONS: Following injection, both hepatocytes and Kupffer cells appeared to be involved in the hepatic clearance and metabolism of both full-length rFVIIa and rFVIIa in complex with at least two plasma protease inhibitors; ATIII and α-2M.

Optimum contribution selection using traditional best linear unbiased prediction and genomic breeding values in aquaculture breeding schemes.

The aim of this study was to compare genetic gain for a traditional aquaculture sib breeding scheme with breeding values based on phenotypic data (TBLUP) with a breeding scheme with genome-wide (GW) breeding values. Both breeding schemes were closed nuclei with discrete generations modeled by stochastic simulation. Optimum contribution selection was applied to restrict pedigree-based inbreeding to either 0.5 or 1% per generation. There were 1,000 selection candidates and a sib test group of either 4,000 or 8,000 fish. The number of selected dams and sires to create full sib families in each generation was determined from the optimum contribution selection method. True breeding values for a trait were simulated by summing the number of each QTL allele and the true effect of each of the 1,000 simulated QTL. Breeding values in TBLUP were predicted from phenotypic and pedigree information, whereas genomic breeding values were computed from genetic markers whose effects were estimated using a genomic BLUP model. In generation 5, genetic gain was 70 and 74% greater for the GW scheme than for the TBLUP scheme for inbreeding rates of 0.5 and 1%. The reduction in genetic variance was, however, greater for the GW scheme than for the TBLUP scheme due to fixation of some QTL. As expected, accuracy of selection increased with increasing heritability (e.g., from 0.77 with a heritability of 0.2 to 0.87 with a heritability of 0.6 for GW, and from 0.53 and 0.58 for TBLUP in generation 5 with sib information only). When the trait was measured on the selection candidate compared with only on sibs and the heritability was 0.4, accuracy increased from 0.55 to 0.69 for TBLUP and from 0.83 to 0.86 for GW. The number of selected sires to get the desired rate of inbreeding was in general less in GW than in TBLUP and was 33 for GW and 83 for TBLUP (rate of inbreeding 1% and heritability 0.4). With truncation selection, genetic gain for the scheme with GW breeding values was nearly twice as large as a scheme with traditional BLUP breeding values. The results indicate that the benefits of applying GW breeding values compared with TBLUP are reduced when contributions are optimized. In conclusion, genetic gain in aquaculture breeding schemes with optimized contributions can increase by as much as 81% by applying genome-wide breeding values compared with traditional BLUP breeding values.

Tissue microarrays compared with whole sections and biochemical analyses. A subgroup analysis of DBCG 82 b&c.

INTRODUCTION: The tissue microarray (TMA) technique comprises the potential of significantly reducing time and tissue spent on slicing and performing immunohistochemical (IHC) stainings of paraffin-embedded tumor tissue. Tissue heterogeneity is an argument against using TMAs, which has been dealt with by increasing the size and number of cores punched from each tumor. No consensus exists on the most optimal size, number, and position of TMA cores in the donor paraffin block and no information exist regarding agreement between TMA cores from two different paraffin blocks from the same tumor or between TMA cores and biochemical analyses. PATIENTS AND METHODS: A central and a peripheral 1mm core and a whole section from each of 54 paraffin blocks from 27 breast cancers included in a one-institution cohort, and a single 1mm central TMA core, from each breast tumor from 1000 patients included in the DBCG82 b&c trials, were IHC stained for ER, PgR and HER2. In addition, ER and PgR were measured in the DBCG82 b&c trials by a biochemical analysis. Statistical analyses included Kappa statistics, Kaplan-Meier survival curves, Log-rank tests, and Cox regression hazards analyses. RESULTS AND CONCLUSION: IHC stainings for ER, PgR, and HER2 showed a substantial agreement between a single 1mm TMA core and the corresponding whole section, between central and peripheral cores, and between cores from two different paraffin blocks from the same tumor. In addition, a fine agreement was found for ER and PgR between IHC stainings of TMA cores and biochemical analyses. Divergence between IHC and biochemical analyses was predominantly due to the chosen thresholds. IHC staining of one 1mm core from each tumor revealed a significant independent prognostic value of PgR and HER2 on overall survival. In conclusion, IHC stainings for ER, PgR, and HER2 of just a single 1mm TMA core seems to be sufficient, as no significant heterogeneity was noticed.

Impact of BCL2 and p53 on postmastectomy radiotherapy response in high-risk breast cancer. A subgroup analysis of DBCG82 b&c.

PURPOSE: To examine p53 and BCL2 expression in high-risk breast cancer patients randomized to postmastectomy radiotherapy (PMRT). PATIENTS AND METHODS: The present analysis included 1 000 of 3 083 high-risk breast cancer patients randomly assigned to PMRT in the DBCG82 b&c studies. Tissue microarray sections were stained with immunohistochemistry for p53 and BCL2. Median potential follow-up was 17 years. Clinical endpoints were locoregional recurrence (LRR), distant metastases (DM), overall mortality, and overall survival (OS). Statistical analyses included Kappa statistics, chi(2) or exact tests, Kaplan-Meier probability plots, Log-rank test, and Cox univariate and multivariate regression analyses. RESULTS: p53 accumulation was not significantly associated with increased overall mortality, DM or LRR probability in univariate or multivariate Cox regression analyses. Kaplan-Meier probability plots showed reduced OS and improved DM and LRR probabilities after PMRT within subgroups of both p53 negative and p53 positive patients. Negative BCL2 expression was significantly associated with increased overall mortality, DM and LRR probability in multivariate Cox regression analyses. Kaplan-Meier probability plots showed a significantly improved overall survival after PMRT for the BCL2 positive subgroup, whereas practically no survival improvement was seen after PMRT for the BCL2 negative subgroup. In multivariate analysis of OS, however, no significant interaction was found between BCL2 and randomization status. Significant reductions in LRR probability after PMRT were recorded within both the BCL2 positive and BCL2 negative subgroups. CONCLUSION: p53 was not associated with survival after radiotherapy in high-risk breast cancer, but BCL2 might be.

Plasma and CSF serpins in Alzheimer disease and dementia with Lewy bodies.

OBJECTIVE: Serine protease inhibitors (serpins), the acute phase reactants and regulators of the proteolytic processing of proteins, have been recognized as potential contributors to the pathogenesis of Alzheimer disease (AD). We measured plasma and CSF levels of serpins in controls and patients with dementia. METHODS: Using rocket immunoelectrophoresis, ELISA, and Luminex xMAP technology, we analyzed plasma levels of alpha(1)-antichymotrypsin and alpha(1)-antitrypsin, and CSF levels of alpha(1)-antichymotrypsin, alpha(1)-antitrypsin, and neuroserpin along with three standard biomarkers (total tau, tau phosphorylated at threonine-181, and the A beta(1-42)) in patients with AD (n = 258), patients with dementia with Lewy bodies (DLB; n = 38), and age-matched controls (n = 37). RESULTS: The level of CSF neuroserpin was significantly higher in AD compared with controls and DLB, whereas CSF alpha(1)-antichymotrypsin and alpha(1)-antitrypsin were significantly higher in both AD and DLB groups than in controls. Results from logistic regression analyses demonstrate a relationship between higher CSF levels of alpha(1)-antichymotrypsin and neuroserpin and increased predicted probability and odds ratios (ORs) of AD (OR 5.3, 95% CI 1.3 to 20.8 and OR 3.3, CI 1.3 to 8.8). Furthermore, a logistic regression model based on CSF alpha(1)-antichymotrypsin, neuroserpin, and A beta(1-42) enabled us to discriminate between AD patients and controls with a sensitivity of 94.7% and a specificity of 77.8%. CONCLUSIONS: Higher CSF levels of neuroserpin and alpha(1)-antichymotrypsin were associated with the clinical diagnosis of Alzheimer disease (AD) and facilitated the diagnostic classification of AD vs controls. CSF serpin levels did not improve the diagnostic classification of AD vs dementia with Lewy bodies.

An approach to derive economic weights in breeding objectives using partial profile choice experiments.

The aim of this study was to show how choice experiments can be used to derive economic weights in breeding objectives. In a choice experiment, respondents are asked to view various alternative descriptions of a good differentiated by their attributes and levels, and are asked to choose their most preferred alternative. Analysis of the data generated can be used to elicit a quantitative description of respondent preference for contrasting attributes and levels. We simulated a partial profile choice experiment with four different attributes (traits) each at three levels. In a partial profile design, the choices are simplified so that only a subset of traits is used for each comparison, making participation in the experimental process less onerous. Three different choice designs were compared. All three designs included four attributes each at three levels where respondents choose between two alternative genotypes. In the first design, respondents choose between two genotypes differing for all four traits simultaneously. In the second and third designs, respondents made choices based on three or two out of the four traits per choice set respectively. The effectiveness of different designs was evaluated based on comparisons between true and simulated preferences for varying numbers of respondents and choice sets per respondent. Choice design and the simulated respondent choice were analysed using a conditional logit model. Regression coefficients from the conditional logit model based on an average of 200 replicated choices across respondents were used to estimate the relative economic weights of traits. A need to account for discounted gene flow principles when formulating the survey questions was emphasised as a critical component of the method. When the relative importance's of four traits were considered, practical designs involving, e.g., 20 choice sets based on a subset of two traits at each choice, and over 30 respondents provided relatively accurate estimates of relative respondent preferences. The method based on a practical choice experiment design can be used to define economic weights for use in animal breeding selection indexes where traditional approaches such as profit equations and bioeconomic models are not practical. The approach may also be of interest to commercial breeding programs wishing to formulate a quantitative understanding of market preferences for attributes of the genestocks that they sell.

A method to define breeding goals for sustainable dairy cattle production.

The objective of this study was to present a method to define breeding goals for sustainable dairy cattle production by adding nonmarket values to market economic values for functional traits in the breeding goal. A nonmarket value can represent the value of improved animal welfare or societal influences for animal production. The nonmarket value for mastitis resistance, conception rate, and stillbirth were derived based on how much farmers or breeding companies were willing to lose in selection response for milk yield to improve functional traits. The desired response for milk yield corresponding to a given percent loss was obtained using desired gain indices. By allowing a 5% loss in the selection response for milk yield, the nonmarket value was found to be 40.4 euro for mastitis resistance, 16.1 euro for conception rate, and 9.7 euro for stillbirth. The nonmarket value increased proportionally with increasing loss in the selection response for milk yield, but the selection response was lower for conception rate than for mastitis resistance because of differences in market economic value and heritability. To increase the response for conception rate, the nonmarket value was also derived for 2 situations, in which the desired responses for milk yield, mastitis resistance, and conception rate were specified. The method can be used to define breeding goals for sustainable production and to increase the response for traits that are at critically low levels. When defining breeding goals for sustainable production, breeding organizations should predict the selection response based on market economic value and add non-market value for traits with unacceptable selection responses.

Preparing and evaluating delivery systems for proteins.

From a formulation perspective proteins are complex and therefore challenging molecules to develop drug delivery systems for. The success of a formulation depends on the ability of the protein to maintain the native structure and activity during preparation and delivery as well as during shipping and long-term storage of the formulation. Therefore, the development and evaluation of successful and promising drug delivery systems is essential. In the present review, some of the particulate drug delivery systems for parenteral delivery of protein are presented and discussed. The challenge for incorporation of protein in particulate delivery systems is exemplified by water-in-oil emulsions.

Derivation of sustainable breeding goals for dairy cattle using selection index theory.

The objective was to present 2 methods for the derivation of nonmarket values for functional traits in dairy cattle using deterministic simulation and selection index theory. A nonmarket value can be a value representing animal welfare and societal influences for animal production, which can be added to market economic values in the breeding goal to define sustainable breeding goals. The first method was restricted indices. A consequence of adding a nonmarket value to a market economic value for a given functional trait is less selection emphasis on milk yield. In the second method, the loss in selection response in milk resulting from greater emphasis on functional traits was quantified. The 2 methods were demonstrated using a breeding goal for dairy cattle with 4 traits (milk yield, mastitis resistance, conception rate, and stillbirth). Nonmarket values derived separately using restricted indices were 0.4 and 2.6 times the value of market economic values for mastitis resistance and conception rate, respectively. Nonmarket values for mastitis resistance and conception rate were both lower when derived simultaneously than when derived separately. This was due to the positive genetic correlation between mastitis resistance and conception rate, and because both traits are negatively correlated with milk yield. Using the second method and accepting a 5% loss in selection response for milk yield, nonmarket values for mastitis, conception rate, and stillbirth were 0.3, 1.4, and 2.9 times the market economic values. It was concluded that the 2 methods could be used to derive nonmarket values for functional traits in dairy cattle.

A simple method to test if the internal mammary lymph nodes are covered by the wide tangent technique in radiotherapy for high-risk breast cancer.

AIM: It is often complicated to include the internal mammary lymph nodes in the radiation field after breast-conserving therapy. Using the wide tangent technique the internal mammary lymph nodes are generally presumed to be included if the medial tangential field border is placed 3 cm across the midline. The current study was designed to test the validity of this assumption, and if possible, to correct the wide tangents without using computed tomography (CT) scanning. PATIENTS AND METHODS: Twenty-one consecutive, high-risk, post-lumpectomy patients were included. An arrangement of three copper wires was mounted in wax placed perpendicular to the skin surface at the ipsilateral border of sternum at intercostal spaces 2 to 4. During a standard simulation for wide tangents, it was examined if the length of the copper wires projected beneath the skin surface (representing the depth of the internal mammary lymph nodes, measured by ultrasound) were included in the wide tangent fields. RESULTS: In only one patient were the internal mammary lymph nodes covered by the wide tangent technique. In 14 of the remaining 20 patients the lateral tangential field border was subsequently moved in the posterior direction, and the internal mammary lymph nodes could be included without unacceptable normal tissue involvement. In the last six patients the irradiated heart and lung volumes exceeded acceptable tolerance levels with this correction, and these patients were referred for three-dimensional CT dose planning. CONCLUSION: The presented simple technique may be helpful if CT scanning is not available. In all other cases CT-based dose plan should ideally be used as a standard in the planning of radiotherapy after breast-conserving surgery to assure optimal inclusion of the relevant target, and to avoid irradiation of large volumes of critical normal tissue.

TR146 cells grown on filters as a model of human buccal epithelium: IV. Permeability of water, mannitol, testosterone and beta-adrenoceptor antagonists. Comparison to human, monkey and porcine buccal mucosa.

The objective of the present study was to evaluate the TR146 cell culture model as an in vitro model of human buccal epithelium. For this purpose, the permeability of water, mannitol and testosterone across the TR146 cell culture model was compared to the permeability across human, monkey and porcine buccal mucosa. Further, the permeability rates of ten beta-adrenoceptor antagonists (acebutolol, alprenolol, atenolol, labetalol, metoprolol, oxprenolol, pindolol, propranolol, timolol and tertatolol) across the TR146 cell culture model and porcine buccal mucosa were related to their lipophilicity (logD(oct; 7.4)) and capacity factor (k') and to their polar water accessible surface area (PWASA). For water, mannitol, testosterone and some of the beta-adrenoceptor antagonists, the permeability enhancement across the TR146 cell culture model in the presence of sodium glycocholate (GC) was determined. The mannitol and testosterone permeability across the TR146 cell culture model could be related to the permeability across porcine and human buccal mucosa. The permeability of the beta-adrenoceptor antagonists across the TR146 cell culture model varied between 2.2 x 10(-6) cm/s (atenolol) and 165 x 10(-6) cm/s (metoprolol). For propranolol the cellular permeability value (P(c)) was lower than expected, probably due to accumulation in the TR146 cell layers. Limited correlation of permeability with k' was observed both for the TR146 cell culture model and the porcine buccal mucosa, although the porcine permeability values were approximately 100 times less than the values determined with the TR146 cell culture model. The permeability values were also found to decrease with increasing PWASA. The PWASA value seemed to be more predictable for permeability than k'. The presence of 12.5 mM GC increased the permeability only for the hydrophilic atenolol, which may help explain the mechanism for GC-induced enhancement. The present results indicate that the TR146 cell culture model can be used as an in vitro model for permeability studies and mechanistic studies of human buccal drug delivery of drugs with different lipophilicity.

TR146 cells grown on filters as a model of human buccal epithelium: III. Permeability enhancement by different pH values, different osmolality values, and bile salts.

The aim of the present study was to evaluate the TR146 cell culture model as an in vitro model of human buccal epithelium with respect to the permeability enhancement by different pH values, different osmolality values or bile salts. For this purpose, the increase in the apparent permeability (P(app)) of the hydrophilic marker mannitol due to exposure to solutions with pH values or osmolality values different from the physiological values was studied. As in studies with solutions of either taurocholate (TC), glycocholate (GC) or glycodeoxycholate (GDC) the results were compared to the increase in P(app) of mannitol obtained in analog studies using porcine buccal mucosa in an Ussing chamber. The effect of the exposure on the electrical resistance of the TR146 cell culture model and the porcine buccal mucosa was measured, and the degree of protein leakage due to GC exposure was investigated in the TR146 cell culture model. The porcine buccal mucosa was approximately ten times less permeable to mannitol than the TR146 cell culture model. The P(app)TC. Increased P(app) values correlated with a decrease in the electrical resistance of the TR146 cell culture model and the porcine buccal mucosa. GC was shown to induce concentration dependent protein leakage in the TR146 cell culture but only from the site of application, and the results indicate that duration of exposure further than 120 min was of minor importance. The present results indicate that the TR146 cell culture model may be a suitable in vitro model for efficacy studies and mechanistic studies of enhancers with potential use in human buccal drug delivery.