Validation of dermatopathological criteria to diagnose cutaneous lesions of mastocytosis: importance of KIT D816V mutation analysis.

BACKGROUND: Cutaneous lesions of mastocytosis (CLM) are often subtle and may require biopsy. However, dermatohistopathological criteria for CLM remain undefined. OBJECTIVES: To establish criteria for CLM by validating histological and molecular parameters. METHODS: In skin samples from Caucasian patients with CLM and controls (atopic dermatitis, chronic urticaria, pruritus, tissue from tumor safety margin excisions), mast cell (MC) numbers, size, shape, distribution, immunostainability with a large panel of markers, pigmentation and presence of KIT D816V mutation were analysed. RESULTS: Forty-seven CLM patients (32 maculopapular cutaneous mastocytosis (MPCM), 15 mastocytomas) and 36 controls were included. Mastocytomas were easily identified by densely packed cuboidal MCs. In MPCM, skin MC density in CD117 stains was higher in CLM patients than in controls (P < 0.0001) and values correlated closely (r = 0.65, P < 0.0001) to results in tryptase stains. The optimized upper dermis cut-off number of 62 MC/mm2 had a sensitivity and specificity of 92% in both stainings, corresponding to approximately 12 MC/high power field (HPF). MC size was larger in MPCM than in controls (P = 0.01). Interstitial (= not perivascular or periadnexal) MCs and stronger basal pigmentation of the epidermis were indicative of MPCM (P < 0.0001 each) and clusters of >3 nucleated MC/HPF exclusively found in MCPM. Surface markers CD2, CD25 and CD30 stained T-lymphocytes, but only negligibly CLM MC. The KIT D816V mutation in formalin fixed paraffin embedded (FFPE) skin was evaluable in 87.5% of MCPM patients and had both 100% sensitivity and specificity. CONCLUSIONS: MPCM can be predicted by major and minor criteria combined in a scoring model. Presence of D816V mutation in FFPE skin and MC density > 27/HPF are >95%-specific major criteria for MPCM. MC densities 12/HPF, interstitial MC, clusters and basal pigmentation are minor criteria.

Effects on feed intake, milk production, and methane emission in dairy cows fed silage or fresh grass with concentrate or fresh grass harvested at early or late maturity stage without concentrate.

The objective of the study was to quantify the effects on dry matter intake (DMI), nutrient digestibility, gas exchange, milk production, and milk quality in dairy cows fed fresh grass harvested at different maturity stages. Sixteen Danish Holstein cows in mid-lactation were divided into 4 blocks and used in 4 incomplete 4 × 2 Latin squares with 2 periods of 21 d. The cows received 1 of 4 treatments in each period, resulting in 8 cows per treatment, as follows: grass-clover silage supplemented with 6 kg/d concentrate pellets (SILc), fresh grass harvested at late maturity stage supplemented with 6 kg/d concentrate pellets (LATc), fresh grass harvested at late maturity stage (LAT), and fresh grass harvested at early maturity stage (ERL). The cows were housed in tiestalls and milked twice daily. The cows had ad libitum access to the forage, and concentrate pellets were divided into equal amounts and fed separately in the morning and afternoon. Fecal samples were collected to determine apparent total-tract digestibility, and samples of rumen fluid were collected for determination of short chain fatty acid composition. Halters were used for measuring eating and rumination time. Gas exchange was measured in open-circuit respiration chambers. Total DMI was higher in LATc and ERL (16.9 ± 0.45 and 15.5 ± 0.39 kg/d, respectively) compared with LAT (14.1 ± 0.42 kg/d). Relative to SILc, cows fed fresh grass experienced a convex pattern in DMI during the experiment. The changes in DMI were related to changes in leaf to stem ratio, fiber concentration, and organic matter digestibility determined in vitro in samples of the fresh grass harvested throughout the experiment. The apparent total-tract digestibility of organic matter was higher in SILc and LAT compared with LATc. Methane yield was lower for LATc compared with LAT (19.5 ± 0.61 vs. 22.6 ± 0.55 g of CH4/kg of DMI), and was not different between LAT and ERL. Compared with LAT, milk yield was higher for ERL (21.1 ± 1.14 vs. 23.4 ± 1.11 kg/d) and energy-corrected milk (ECM) yield was higher for LATc (21.5 ± 0.99 vs. 25.3 ± 1.03 kg/d). We detected no differences in milk or ECM yield between SILc and LATc. Milk protein yield was higher and milk fat concentration was lower in LATc compared with LAT. The fatty acid percentages of ∑C4-C14:1 and ∑C16 in milk were higher for SILc compared with LATc, signifying pronounced de novo synthesis. The n-6:n-3 ratio in milk fatty acids was lower for SILc and LAT compared with LATc, indicating improved nutritional quality for SILc and LAT. However, retinol concentration in milk was lower in SILc compared with all other treatments. The study implies that feeding silage instead of fresh grass has no effect on DMI, ECM yield, or CH4 yield, and that concentrate supplementation can increase milk production, affects milk quality, and reduces the effect on climate, whereas feeding less mature grass increases DMI and milk yield, but has no effect on CH4 yield.

Clinically decisive (dis)agreement in multidisciplinary team assessment of esophageal squamous cell carcinoma; a prospective, national, multicenter study.

BACKGROUND: Decisions regarding tumor staging, operability, resectability, and treatment strategy in patients with esophageal cancer are made at multidisciplinary team (MDT) conferences. We aimed to assess interobserver agreement from four national MDT conferences and whether this would have a clinical impact. METHODS: A total of 20 patients with esophageal cancer were included across all four upper gastrointestinal (GI) cancer centers. Fully anonymized patient data were distributed among the MDT conferences which decided on TNM category, resectability, operability, curability, and treatment strategy blinded to each other's decisions. The interobserver agreement was expressed as both the raw observer agreement and with Krippendorff's α values. Finally, a case-by-case evaluation was performed to determine if disagreement would have had a clinical impact. RESULTS: A total of 80 MDT evaluations were available for analysis. A moderate to near-perfect observer agreement of 79.2%, 55.8%, and 82.5% for TNM category was observed, respectively. Substantial agreement for resectability and moderate agreement for curability were found. However, an only fair agreement was observed for the operability category. The treatment strategies had a slight agreement which corresponded to disagreement having a clinical impact in 12 patients. CONCLUSIONS: Esophageal cancer MDT conferences had an acceptable interobserver agreement on resectability and TM categories; however, the operability assessment had a high level of disagreement. Consequently, the agreement on treatment strategy was reduced with a potential clinical impact. In future MDT conferences, emphasis should be on prioritizing the relevant information being readily available (operability, T & M categories) to minimize the risk of disagreement in the assessments and treatment strategies, and thus, delayed or suboptimal treatment.

Basic symptoms influence real-life functioning and symptoms in individuals at high risk for psychosis.

OBJECTIVE: To investigate potential clinical differences in high-risk profiles presenting with and without basic symptoms, and additionally investigate the association between basic symptoms and clinical symptoms, functioning, and cognition. METHODS: High-risk individuals (n = 133) were stratified into individuals fulfilling ultra-high-risk (UHR) criteria (n = 59) and individuals fulfilling UHR+ basic symptoms criteria (BS) (n = 74). Group differences were assessed on clinical symptoms, real-life functioning, and cognition. Regression analyses were conducted to elucidate on the relationship between BS and clinical symptoms, functioning, neurocognition, and social cognition. RESULTS: The group fulfilling both UHR+ BS criteria had significantly more symptoms and lower real-life functioning and quality of life but not more cognitive deficits. BS influenced on attenuated psychotic, depressive, and general symptoms, but only modestly on negative symptoms. No relationship between BS and neuro- and social cognition was established except for an association with emotion recognition processing speed. BS influenced real-life functioning, and this finding was sustained when controlling for the effect of negative symptoms. CONCLUSIONS: Our findings indicate that BS contribute highly to the distress and symptom load of clinical high-risk individuals. Longitudinal findings are needed to establish the predictive validity of BS on high-risk individuals' clinical and functional prognosis.

Effects of an individual cow concentrate strategy on production and behavior.

The aim of this study was to investigate the effects of a concentrate strategy based on the individual cow partial mixed ration (PMR) intake compared with a flat rate. Half of the cows were fed the individual concentrate strategy, whereas the other half were fed the control strategy. The individually fed cows were offered a concentrate proportion equal to 30% of the ad libitum intake of the PMR in the automatic milking system, and the control cows were offered 3 kg of concentrate/d in the automatic milking system. The cows (83 Holstein and 64 Jersey), kept in 2 separate groups, were blocked between the treatments according to expected calving date, breed, and parity and were randomly divided between treatments. Lactation was divided into 3 periods (early, mid, and late lactation), and the MIXED and GLIMMIX procedures in SAS (SAS Institute Inc., Cary, NC) were used to analyze the production and behavioral responses. The response trajectories during lactation were analyzed with a MIXED procedure fitted by Wilmink parameters. The individually fed cows had a higher concentrate intake and a lower PMR intake than control cows. Moreover, the total dry matter intake and energy-corrected milk yield did not differ between concentrate strategies. The actual average concentrate intake reached 19% of the PMR intake during mid lactation and not 30% of the PMR intake, as intended with the strategy. This was due to leftovers and lower allocation than intended. The cow behavior did not differ between concentrate strategies. However, variation in PMR intake and eating rate between cows fed individually was lower during mid lactation and lower for lying bouts during early lactation compared with control cows. This may indicate that an individual cow concentrate strategy results in a more stable PMR intake and time budget, which potentially could improve welfare, but this needs further investigation. The overall conclusion is that the cows were robust to adjustments in the concentrate allowance.

Herd factors influencing free fatty acid concentrations in bulk tank milk.

Free fatty acid (FFA) concentrations can be elevated in raw milk due to improper handling and management at the dairy farm, and high concentrations of FFA can lead to off flavors in milk. This study aimed to describe how the herd production system, milking system, feeding and technological factors impact on FFA concentrations in bulk tank milk. FFA concentrations in bulk milk samples from 259 organic and 3326 conventional herds were analyzed by FT-IR during one year. The FFA content was significantly lower in bulk milk from organic than conventional herds. This was most evident during the summer half-year when the organic cows graze pasture. Bulk milk from automatic milking systems (AMS) and tie-stalls contained greater concentrations of FFA than any other milking parlor systems. In AMS, high milking frequency was found to be the most significant contributor to elevated FFA content in milk. Moreover, a strong interaction was found between milking interval and production system (organic vs. conventional). The technical factors, pre-cooling, onset time for cooling after milk inlet, contact between milk and agitation also impacted on the FFA concentration, whereas other technical factors including centrifugal pump type, length and height of pumping line and type of AMS manufacturer were found to be without significant effect on FFA. Feeding variables, based on feeding plans and evaluation, only explained a small part of the variation in bulk milk FFA. Overall, this study demonstrated that AMS compared to other milking system contributes significantly to increased FFA concentration in bulk tank milk, and within AMS high milking frequency contributes to increased FFA concentration.

Omalizumab prevents anaphylaxis and improves symptoms in systemic mastocytosis: Efficacy and safety observations.

BACKGROUND: Patients with systemic mastocytosis (SM) may suffer from mast cell (MC) mediator-related symptoms insufficiently controlled by conventional therapy. Omalizumab is an established treatment in other MC-driven diseases, but experiences in SM are limited. OBJECTIVE: To assess the efficacy and safety of omalizumab in SM. METHODS: In our patient cohort, we evaluated all SM patients treated with omalizumab. A physician global assessment of type and severity of symptoms was performed at baseline, at 3 and 6 months and at latest follow-up. Quality of life was assessed by visual analogue scale. S-tryptase and KIT D816V allele burden were monitored. RESULTS: A total of 14 adult SM patients (10 ISM, 2 BMM, 1 SSM, and 1 ASM-AHN) received omalizumab with a median duration of 17 months (range: 1-73 months). One patient was excluded due to concomitant cytoreductive therapy. In the remaining 13 patients, we observed a significant reduction in symptoms, with complete symptom control in five (38.5%), major response in three (23.1%), and a partial response in three (23.1%) patients, whereas two patients (15.4%) withdrew due to subjective side-effects at first dose. The treatment was most effective for recurrent anaphylaxis and skin symptoms, less for gastrointestinal, musculoskeletal, and neuropsychiatric symptoms. Patient-reported quality of life showed significant improvement. No significant changes in s-tryptase/KIT D816V allele burden were observed. No severe adverse events were recorded. CONCLUSIONS: Omalizumab appears to be a promising treatment option in SM, effectively preventing anaphylaxis and improving chronic MC mediator-related symptoms, insufficiently controlled by conventional therapy. Controlled studies are needed to substantiate findings.

Laboratory maintenance does not alter ecological and physiological patterns among species: a Drosophila case study.

Large comparative studies in animal ecology, physiology and evolution often use animals reared in the laboratory for many generations; however, the relevance of these studies hinges on the assumption that laboratory populations are still representative for their wild living conspecifics. In this study, we investigate whether laboratory-maintained and freshly collected animal populations are fundamentally different and whether data from laboratory-maintained animals are valid to use in large comparative investigations of ecological and physiological patterns. Here, we obtained nine species of Drosophila with paired populations of laboratory-maintained and freshly collected flies. These species, representing a range of ecotypes, were assayed for four stress-tolerance, two body-size traits and six life-history traits. For all of these traits, we observed small differences in species-specific comparisons between field and laboratory populations; however, these differences were unsystematic and laboratory maintenance did not eclipse fundamental species characteristics. To investigate whether laboratory maintenance influence the general patterns in comparative studies, we correlated stress tolerance and life-history traits with environmental traits for the laboratory-maintained and freshly collected populations. Based on this analysis, we found that the comparative physiological and ecological trait correlations are similar irrespective of provenience. This finding is important for comparative biology in general because it validates comparative meta-analyses based on laboratory-maintained populations.

Prospective evaluation of the diagnostic value of sensitive KIT D816V mutation analysis of blood in adults with suspected systemic mastocytosis.

BACKGROUND: Sensitive KIT D816V mutation analysis of blood has been proposed to guide bone marrow (BM) investigation in suspected systemic mastocytosis (SM). The aim of this prospective study was for the first time to compare the D816V status of the "screening blood sample" used to guide BM biopsy in suspected SM to the outcome of the subsequent BM investigation. METHODS: Fifty-eight adult patients with suspected SM were included. The outcome of sensitive KIT D816V analysis of blood was compared to the result of the BM investigation. RESULTS: Screening blood samples from 44 of 58 patients tested D816V-positive. In 43 of these, SM was subsequently diagnosed in the BM investigation. One patient with a D816V-positive screening sample was diagnosed with monoclonal MC activation syndrome. Screening blood samples from 14 patients tested D816V-negative. SM was subsequently diagnosed in five of these, whereas nine patients did not fulfill any diagnostic SM criteria (excluding tryptase criterion). Of the 48 SM patients, 90% tested D816V-positive. Thirteen SM patients presented with Hymenoptera venom-induced anaphylaxis, no skin lesions, and baseline serum tryptase ≤20 ng/mL. Of these, 92% tested D816V-positive in the screening blood sample. CONCLUSION: This prospective study demonstrates that a D816V-positive result in a screening blood sample identifies SM among patients with hymenoptera venom-induced anaphylaxis in whom the diagnosis would most probably have been missed, with potential severe implications. The observed false-negative screening results also underline that BM investigation is mandatory in all adult patients with clear signs of, or highly suspected SM, regardless of the KIT mutation status.

Patterns of anaphylaxis after diagnostic workup: A follow-up study of 226 patients with suspected anaphylaxis.

BACKGROUND: Most published studies on anaphylaxis are retrospective or register based. Data on subsequent diagnostic workup are sparse. We aimed to characterize patients seen with suspected anaphylaxis at the emergency care setting (ECS), after subsequent diagnostic workup at our Allergy Center (AC). METHODS: Prospective study including patients from the ECS, Odense University Hospital, during May 2013-April 2014. Possible anaphylaxis cases were daily identified based on a broad search profile including history and symptoms in patient records, diagnostic codes and pharmacological treatments. At the AC, all patients were evaluated according to international guidelines. RESULTS: Among 226 patients with suspected anaphylaxis, the diagnosis was confirmed in 124 (54.9%) after diagnostic workup; 118 of the 124 fulfilled WAO/EAACI criteria of anaphylaxis at the ECS, while six were found among 46 patients with clinical suspicion but not fulfilling the WAO/EAACI criteria at the ECS. The estimated incidence rate of anaphylaxis was 26 cases per 100 000 person-years and the one-year period prevalence was 0.04%. The most common elicitor was drugs (41.1%) followed by venom (27.4%) and food (20.6%). In 13 patients (10.5%), no elicitor could be identified. Mastocytosis was diagnosed in 7.7% of adult patients and was significantly associated with severe anaphylaxis. Atopic diseases were significantly associated only with food-induced anaphylaxis. Cofactors were present in 58.1% and were significantly associated with severe anaphylaxis. CONCLUSION: A broad search profile in the ECS and subsequent diagnostic workup is important for identification and classification of patients with anaphylaxis. Evaluation of comorbidities and cofactors is important.

Negative symptoms mediate the relationship between neurocognition and function in individuals at ultrahigh risk for psychosis.

OBJECTIVE: Neurocognition is known to impact functioning in individuals at ultrahigh risk (UHR) for psychosis, but studies investigating potential mediators of this relationship are scarce. Building on evidence from schizophrenia spectrum disorders, the study tested whether negative symptoms and social skills act as mediators between neurocognition and functional outcome in UHR individuals. METHODS: Ultrahigh risk participants (N = 84) underwent neurocognitive testing using the Brief Assessment of Cognition in Schizophrenia. Social skills and negative symptoms were assessed using the High-Risk Social Challenge task and the Scale for the Assessment of Negative Symptoms respectively. Four instruments were used to assess overall functioning, and one instrument assessed quality of life encompassing social functioning. RESULTS: The cross-sectional analyses revealed that neurocognition was related to the measures of functioning. Negative symptoms mediated the relationship between neurocognition and four of the five measures of functioning. We did not find social skills to mediate between neurocognition and functioning. CONCLUSION: Negative symptoms appear to mediate the relationship between neurocognition and functional outcome in UHR individuals, but the finding needs to be confirmed and extended to longitudinal studies. This underscores the importance of focusing on both neurocognition and negative symptoms when aiming at improving the functional outcome of UHR individuals.

Early lactation production, health, and welfare characteristics of cows selected for extended lactation.

Some cows are able to achieve relatively high milk yields during extended lactations beyond 305 d in milk, and farmers may be able to use this potential by selecting the most suitable cows for an extended lactation. However, the decision to postpone insemination has to rely on information available in early lactation. The main objectives of this study were, therefore, to assess the association between the information available in early lactation and the relative milk production of cows on extended lactation, and to investigate if this information can be used to differentiate time of first insemination between cows. Data came from 4 Danish private herds practicing extended lactation in which some cows are selected to have a delayed time of planned first insemination. Average herd size varied from 93 to 157 cows, and milk yield varied from 7,842 to 12,315 kg of energy-corrected milk (ECM) per cow per year across herds. The analysis was based on 422 completed extended lactations (427 ± 87 d), and each lactation was assigned to 1 of 3 (low, medium, and high) milk performance groups (MPG) within parity group within herd based on a standardized lactation yield. For cows in the high MPG, peak ECM yield, and ECM yield at dry off were significantly greater, the relative reduction in milk yield between 60 and 305 d in milk was significantly smaller, and a smaller proportion had a body condition score (scale: 1-5) at dry off of 3.5 or greater compared with cows in low MPG. Previous lactation days in milk at peak ECM yield and ECM yield at dry off were higher, the relative reduction in milk yield between 60 and 305 d in milk was smaller, and the number of inseminations per conception was higher for multiparous cows in high MPG compared with low. Current lactation ECM yield at second and third milk recording were greater for cows in high MPG compared with low. A principal component analysis indicated that variables related to fertility, diseases, and milk yield explained most of the total variation between primiparous cows, whereas variables related to milk yield, fertility, and days in milk at peak yield were the most dominating for multiparous cows. Our study indicated that milk yields in previous lactation and at second and third milk recording correlate well with milk production potential, and therefore, may be promising indicators when selecting the most suitable cows for extended lactation.

Effect of calving interval and parity on milk yield per feeding day in Danish commercial dairy herds.

The idea of managing cows for extended lactations rather than lactations of the traditional length of 1 yr primarily arose from observations of increasing problems with infertility and cows being dried off with high milk yields. However, it is vital for the success of extended lactation practices that cows are able to maintain milk yield per feeding day when the length of the calving interval (CInt) is increased. Milk yield per feeding day is defined as the cumulated lactation milk yield divided by the sum of days between 2 consecutive calvings. The main objective of this study was to investigate the milk production of cows managed for lactations of different lengths, and the primary aim was to investigate the relationship between CInt, parity, and milk yield. Five measurements of milk yield were used: energy-corrected milk (ECM) yield per feeding day, ECM yield per lactating day, cumulative ECM yield during the first 305 d of lactation, as well as ECM yield per day during early and late lactation. The analyses were based on a total of 1,379 completed lactations from cows calving between January 2007 and May 2013 in 4 Danish commercial dairy herds managed for extended lactation for several years. Herd-average CInt length ranged from 414 to 521 d. The herds had Holstein, Jersey, or crosses between Holstein, Jersey, and Red Danish cows with average milk yields ranging from 7,644 to 11,286 kg of ECM per cow per year. A significant effect of the CInt was noted on all 5 measurements of milk yield, and this effect interacted with parity for ECM per feeding day, ECM per lactating day and ECM per day during late lactation. The results showed that cows were at least able to produce equivalent ECM per feeding day with increasing CInt, and that first- and second-parity cows maintained ECM per lactating day. Cows with a CInt between 17 and 19 mo produced 476 kg of ECM more during the first 305 d compared with cows with a CInt of less than 13 mo. Furthermore, early-lactation ECM yield was greater for all cows and late-lactation ECM yield was less for second-parity and older cows when undergoing an extended compared with a shorter lactation. Increasing CInt increased the dry period length with 3 to 5d. In conclusion, the group of cows with longer CInt were able to produce at least equivalent amounts of ECM per feeding day when the CInt was up to 17 to 19 mo on these 4 commercial dairy farms.

Inbreeding depression across a nutritional stress continuum.

Many natural populations experience inbreeding and genetic drift as a consequence of nonrandom mating or low population size. Furthermore, they face environmental challenges that may interact synergistically with deleterious consequences of increased homozygosity and further decrease fitness. Most studies on inbreeding-environment (I-E) interactions use one or two stress levels, whereby the resolution of the possible stress and inbreeding depression interaction is low. Here we produced Drosophila melanogaster replicate populations, maintained at three different population sizes (10, 50 and a control size of 500) for 25 generations. A nutritional stress gradient was imposed on the replicate populations by exposing them to 11 different concentrations of yeast in the developmental medium. We assessed the consequences of nutritional stress by scoring egg-to-adult viability and body mass of emerged flies. We found: (1) unequivocal evidence for I-E interactions in egg-to-adult viability and to a lesser extent in dry body mass, with inbreeding depression being more severe under higher levels of nutritional stress; (2) a steeper increase in inbreeding depression for replicate populations of size 10 with increasing nutritional stress than for replicate populations of size 50; (3) a nonlinear norm of reaction between inbreeding depression and nutritional stress; and (4) a faster increase in number of lethal equivalents in replicate populations of size 10 compared with replicate populations of size 50 with increasing nutritional stress levels. Our data provide novel and strong evidence that deleterious fitness consequences of I-E interactions are more pronounced at higher nutritional stress and at higher inbreeding levels.

Feeding, production, and efficiency of Holstein-Friesian, Jersey, and mixed-breed lactating dairy cows in commercial Danish herds.

The objective of this paper was to compare efficiency measures, milk production, and feed intake for lactating cows in commercial herds using different breeds and production and milking systems. To accomplish this, we used all feed evaluations made by the Danish extension service during the period November 2012 to April 2013 for 779 herds, of which 508 were Holstein-Friesian (HOL); 100 were Jersey (JER); and 171 herds were a mixture of these 2 breeds, other dairy breeds, and crossbreeds (OTH). The annually recorded, herd-average energy-corrected milk (ECM) yield was 8,716kg (JER) and 9,606kg (HOL); and average herd size was 197 cows (HOL) and 224 cows (JER). All cows were fed a total mixed or partial mixed ration supplemented with concentrate from feeding stations, housed in loose housing systems with a slatted floor, and milked in either a parlor milking unit or an automatic milking system. Energy efficiency was calculated as net energy efficiency defined as total energy demand as a percentage of energy intake and as residual feed intake defined as energy intake (net energy for lactation; NEL) minus energy requirement. Production efficiency was expressed as kilograms of ECM per kilogram of dry matter intake (DMI), kilograms of ECM per 10 MJ of net energy intake (NEL), kilograms of ECM per 100kg of BW, and kilograms of DMI per 100kg of BW. Environmental efficiency was expressed by the nitrogen efficiency calculated as N in milk and meat as a percentage of N in intake, and as enteric emission of methane expressed as kilograms of ECM per megajoule of CH4. Mean milk yield for lactating cows was 30.4kg of ECM in HOL and 3kg less in JER, with OTH herds in between. Mean NEL intake was 122 MJ in JER, increasing to 147 MJ in HOL, whereas ration energy density between breeds did not differ (6.4-6.5 MJ of NEL per kg of DMI). The NEL intake and DMI explained 56 and 47%, respectively, of variation in production (ECM) for HOL herds but only 44 and 27% for JER. Jersey had a higher efficiency than HOL and OTH, except in nitrogen efficiency, where no significant difference between breeds existed. Most of the efficiency measures were internally significantly correlated and in general highly positively correlated with milk production, whereas the correlation to DMI was less positive and for JER negative for net energy efficiency, kilograms of ECM per kilogram of DMI, and nitrogen efficiency. Only little of the variation in efficiency between herds could be explained by differences in nutrient or roughage content of DMI. This could be explained by the fact that data were collected from herds purchasing feed planning and evaluation from the extension service.

Proposed diagnostic algorithm for patients with suspected mastocytosis: a proposal of the European Competence Network on Mastocytosis.

Mastocytosis is an emerging differential diagnosis in patients with more or less specific mediator-related symptoms. In some of these patients, typical skin lesions are found and the diagnosis of mastocytosis can be established. In other cases, however, skin lesions are absent, which represents a diagnostic challenge. In the light of this unmet need, we developed a diagnostic algorithm for patients with suspected mastocytosis. In adult patients with typical lesions of mastocytosis in the skin, a bone marrow (BM) biopsy should be considered, regardless of the basal serum tryptase concentration. In adults without skin lesions who suffer from mediator-related or other typical symptoms, the basal tryptase level is an important parameter. In those with a slightly increased tryptase level, additional investigations, including a sensitive KIT mutation analysis of blood leucocytes or measurement of urinary histamine metabolites, may be helpful. In adult patients in whom (i) KIT D816V is detected and/or (ii) the basal serum tryptase level is clearly increased (>25-30 ng/ml) and/or (iii) other clinical or laboratory features suggest the presence of 'occult' mastocytosis or another haematologic neoplasm, a BM investigation is recommended. In the absence of KIT D816V and other signs or symptoms of mastocytosis or another haematopoietic disease, no BM investigation is required, but the clinical course and tryptase levels are monitored in the follow-up. In paediatric patients, a BM investigation is usually not required, even if the tryptase level is increased. Although validation is required, it can be expected that the algorithm proposed herein will facilitate the management of patients with suspected mastocytosis and help avoid unnecessary referrals and investigations.

A Drosophila laboratory evolution experiment points to low evolutionary potential under increased temperatures likely to be experienced in the future.

The ability to respond evolutionarily to increasing temperatures is important for survival of ectotherms in a changing climate. Recent studies suggest that upper thermal limits may be evolutionary constrained. We address this hypothesis in a laboratory evolution experiment, encompassing ecologically relevant thermal regimes. To examine the potential for species to respond to climate change, we exposed replicate populations of Drosophila melanogaster to increasing temperatures (0.3 °C every generation) for 20 generations, whereas corresponding replicate control populations were held at benign thermal conditions throughout the experiment. We hypothesized that replicate populations exposed to increasing temperatures would show increased resistance to warm and dry environments compared with replicate control populations. Contrasting replicate populations held at the two thermal regimes showed (i) an increase in desiccation resistance and a decline in heat knock-down resistance in replicate populations exposed to increasing temperatures, (ii) similar egg-to-adult viability and fecundity in replicate populations from the two thermal regimes, when assessed at high stressful temperatures and (iii) no difference in nucleotide diversity between thermal regimes. The limited scope for adaptive evolutionary responses shown in this study highlights the challenges faced by ectotherms under climate change.

The effects of sustained aerobic swimming on osmoregulatory pathways in Atlantic salmon Salmo salar smolts.

Atlantic salmon Salmo salar smolts were exposed to one of the four different aerobic exercise regimens for 10 weeks followed by a 1 week final smoltification period in fresh water and a subsequent eight-day seawater transfer period. Samples of gill and intestinal tissue were taken at each time point and gene expression was used to assess the effects of exercise training on both branchial and intestinal osmoregulatory pathways. Real-time polymerase chain reaction (PCR) analysis revealed that exercise training up-regulated the expression of seawater relevant genes in the gills of S. salar smolts, including Na(+) , K(+) ATPase (nka) subunit α1b, the Na(+) , K(+) , 2 Cl(-) co-transporter (nkcc1) and cftr channel. These findings suggest that aerobic exercise stimulates expression of seawater ion transport pathways that may act to shift the seawater transfer window for S. salar smolts. Aerobic exercise also appeared to stimulate freshwater ion uptake mechanisms probably associated with an osmorespiratory compromise related to increased exercise. No differences were observed in plasma Na(+) and Cl(-) concentrations as a consequence of exercise treatment, but plasma Na(+) was lower during the final smoltification period in all treatments. No effects of exercise were observed for intestinal nkcc2, nor the Mg(2+) transporters slc41a2 and transient receptor protein M7 (trpm7); however, expression of both Mg(2+) transporters was affected by salinity transfer suggesting a dynamic role in Mg(2+) homeostasis in fishes.

A rapid qPCR method for genetic sex identification of Salmo salar and Salmo trutta including simultaneous elucidation of interspecies hybrid paternity by high-resolution melt analysis.

This study presents an improved duplex quantitative polymerase chain reaction (qPCR) method using the master sex-determining gene sdY as a marker for simultaneous genetic sex identification of salmonids of the Salmo genus and paternity elucidation for Salmo salar × Salmo trutta hybrids. This method will provide a new, simple and economical molecular tool for ecological studies of these species as well as for aquaculture purposes.

Long-term hypo-osmoregulatory capacity in downstream migrating Atlantic salmon Salmo salar L. smolts.

The duration of hypo-osmoregulatory capacity in downstream migrating Atlantic salmon Salmo salar L smolts previously stocked as startfed young-of-the year (YOY) parr was tested in the River Dalåa from mid-May to late-June 1999. Hypo-osmoregulatory capacity, measured as plasma osmolality and chloride, was assessed after seawater (SW) challenge tests (168 h, salinity = 35). All S. salar exhibited sufficient hypo-osmoregulatory capacity at the initiation of downstream migration in mid-May. Migrating S. salar smolts caught in mid-May and retained in fresh water displayed no signs of de-smoltification as they maintained hypo-osmoregulatory capacity through June. This indicates a physiological smolt window that lasts a minimum of 6 weeks (330 degree days; D°) for hatchery-produced S. salar smolts stocked as YOY parr. Based on the observed river migration speeds, it can be assumed that the S. salar smolts entered SW 2-4 weeks after initiation of migration in the upper parts of the River Dalåa. Hence, based on smolt migration and SW tolerance, it is suggested that stocking of YOY parr is a viable enhancement strategy in the River Dalåa.