Inter- and intra-observer variation in phytolith morphometry.

BACKGROUND: Archaeobotanists and palaeoecologists extensively use geometric morphometrics to identify plant opal phytoliths. Particularly when applied to assemblages of phytoliths from concentrations retrieved from closed contexts, morphometric data from archaeological phytoliths compared with similar data from reference material may allow taxonomic attribution. Observer variation is one aspect of phytolith morphometry that has received little attention but may be an important source of error, and hence cause of potential misidentification of plant remains. SCOPE: To investigate inter- and intra-observer variation in phytolith morphometry, eight researchers (observers) from different laboratories measured 50 samples each from three phytolith morphotypes, Bilobate, Bulliform flabellate and Elongate dendritic, three times, under the auspices of the International Committee for Phytolith Morphometrics (ICPM). METHODS: Data for 17 size and shape variables were collected for each phytolith by manually digitising a phytolith outline (mask) from a photograph, followed by measurement of the mask with open-source morphometric software. KEY RESULTS: Inter-observer variation ranged from 0 to 23% difference from the mean of all observers. Intra-observer variation ranged from 0 to 9% difference from the mean of individual observers per week. Inter- and intra-observer variation was generally higher among inexperienced researchers. CONCLUSIONS: Scaling errors were a major cause of variation and occurred more with less experienced researchers, which is likely related to familiarity with data collection. The results indicate that inter- and intra-observer variation can be substantially reduced by providing clear instructions for and training with the equipment, photo capturing, software, data collection and data cleaning. In this paper, the ICPM provides recommendations to minimise variation.Advances in automatic data collection may eventually reduce inter- and intra-observer variation, but until this is common practice, the ICPM recommends that phytolith morphometric analyses adhere to standardised guidelines to assure that measured phytolith variables are accurate, consistent and comparable between different researchers and laboratories.

Simultaneous Inference of Multiple Binary Endpoints in Biomedical Research: Small Sample Properties of Multiple Marginal Models and a Resampling Approach.

In biomedical research, the simultaneous inference of multiple binary endpoints may be of interest. In such cases, an appropriate multiplicity adjustment is required that controls the family-wise error rate, which represents the probability of making incorrect test decisions. In this paper, we investigate two approaches that perform single-step p $p$ -value adjustments that also take into account the possible correlation between endpoints. A rather novel and flexible approach known as multiple marginal models is considered, which is based on stacking of the parameter estimates of the marginal models and deriving their joint asymptotic distribution. We also investigate a nonparametric vector-based resampling approach, and we compare both approaches with the Bonferroni method by examining the family-wise error rate and power for different parameter settings, including low proportions and small sample sizes. The results show that the resampling-based approach consistently outperforms the other methods in terms of power, while still controlling the family-wise error rate. The multiple marginal models approach, on the other hand, shows a more conservative behavior. However, it offers more versatility in application, allowing for more complex models or straightforward computation of simultaneous confidence intervals. The practical application of the methods is demonstrated using a toxicological dataset from the National Toxicology Program.

Optimization of Enzymatic and Chemical Decellularization of Native Porcine Heart Valves for the Generation of Decellularized Xenografts.

One of the most important medical interventions for individuals with heart valvular disease is heart valve replacement, which is not without substantial challenges, particularly for pediatric patients. Due to their biological properties and biocompatibility, natural tissue-originated scaffolds derived from human or animal sources are one type of scaffold that is widely used in tissue engineering. However, they are known for their high potential for immunogenicity. Being free of cells and genetic material, decellularized xenografts, consequently, have low immunogenicity and, thus, are expected to be tolerated by the recipient's immune system. The scaffold ultrastructure and ECM composition can be affected by cell removal agents. Therefore, applying an appropriate method that preserves intact the structure of the ECM plays a critical role in the final result. So far, there has not been an effective decellularization technique that preserves the integrity of the heart valve's ultrastructure while securing the least amount of genetic material left. This study demonstrates a new protocol with untraceable cells and residual DNA, thereby maximally reducing any chance of immunogenicity. The mechanical and biochemical properties of the ECM resemble those of native heart valves. Results from this study strongly indicate that different critical factors, such as ionic detergent omission, the substitution of Triton X-100 with Tergitol, and using a lower concentration of trypsin and a higher concentration of DNase and RNase, play a significant role in maintaining intact the ultrastructure and function of the ECM.

QTL mapping and transcriptome analysis identify novel QTLs and candidate genes in Brassica villosa for quantitative resistance against Sclerotinia sclerotiorum.

KEY MESSAGE: Novel QTLs and candidate genes for Sclerotinia-resistance were identified in B. villosa, a wild Brassica species, which represents a new genetic source for improving oilseed rape resistance to SSR. Sclerotinia stem rot (SSR), caused by Sclerotinia sclerotiorum, is one of the most destructive diseases in oilseed rape growing regions. To date, there is no effective genetic resistance against S. sclerotiorum in the B. napus germplasm and knowledge of the molecular plant-fungal interaction is also limited. To identify new resistance resources, we screened a set of wild Brassica species and identified B. villosa (BRA1896) with a high level of Sclerotinia-resistance. Two segregating F2 populations for Sclerotinia-resistance, generated by interspecific crosses between the resistant B. villosa (BRA1896) and the wild susceptible B. oleracea (BRA1909) were assessed for Sclerotinia-resistance. Genetic mapping using a 15-k Illumina Infinium SNP-array resulted in a high-density genetic map containing 1,118 SNP markers and spanning a total genetic length of 792.2 cM. QTL analysis revealed seven QTLs explaining 3.8% to 16.5% of phenotypic variance. Intriguingly, RNAseq-based transcriptome analysis identified genes and pathways specific to B. villosa, of which a cluster of five genes encoding putative receptor-like kinases (RLKs) and two pathogenesis-related (PR) proteins are co-localized within a QTL on chromosome C07. Furthermore, transcriptomic analysis revealed enhanced ethylene (ET)-activated signaling in the resistant B. villosa, which is associated with a stronger plant immune response, depressed cell death, and enhanced phytoalexin biosynthesis compared to the susceptible B. oleracea. Our data demonstrates that B. villosa represents a novel and unique genetic source for improving oilseed rape resistance against SSR.

The thermal dependence of the protein-sparing effect in rainbow trout (Oncorhynchus mykiss, Walbaum 1792).

The authors performed an instantaneous bioenergetic study with rainbow trout (Oncorhynchus mykiss) of 206.3 g ± 2.9 g in a group respirometer of nine 250 l tanks at five different water temperatures (12, 14, 16, 18, 20°C) to determine the optimal thermal condition for a maximal visualization of the protein-sparing effect. Twelve fish per tank were tested at a stocking density of 9.94 kg m-3 ± 0.14 kg m-3 and fed three low-protein/high-energy diets with constant crude protein content of c. 35% and three different energy contents (17.35, 18.76, 20.50 MJ kg-1 ) once daily at a ration of 1.3% body weight (n = 3). Energy levels were increased by adding gelatinized wheat starch as a carbohydrate source and fish oil, canola oil and palmitin as lipid sources. Three different dietary digestible protein/digestible energy ratios (DP/DE: 20.38, 19.08, 18.09 mg kJ-1 ) were achieved by replacing bentonite as a non-nutritive filler with carbohydrates and lipids. Oxygen consumption and ammonia excretion were assessed to obtain the potentially retainable energy (RE) and ammonia quotient (AQ) as benchmarks for potential growth and protein-sparing effect. The results showed the lowest relative metabolic combustion of protein at 16.9°C ± 0.1°C. The authors determined this temperature to set the optimal thermal condition for the induction of a maximum protein-sparing effect in juvenile rainbow trout. Increasing the DP/DE ratio significantly altered the magnitude of the relative metabolic protein use but had no effect on its interactions with temperature. The authors were able to reduce average metabolic fuel use of protein across diets from 16.2% ± 2.3% at 12°C to 8.0% ± 1.2% at 16°C. This study found no relevant significant differences of RE with the environmental temperature.

Early prediction of final infarct volume with material decomposition images of dual-energy CT after mechanical thrombectomy.

PURPOSE: Evaluation of water material density images (wMDIm) of dual-energy CT (DECT) for earlier prediction of final infarct volume (fiV) in follow-up single-energy CT (SECT) and correlation with clinical outcome. METHODS: Fifty patients (69 years, ± 12.1, 40-90, 50% female) with middle cerebral artery (MCA) occlusions were included. Early infarct volumes were analyzed in monoenergetic images (MonoIm) and wMDIm at 60 keV and compared with the fiV in SECT 4.9 days (± 4) after thrombectomy. Association between infarct volume and functional outcome was tested by linear regression analysis. RESULTS: wMDIm shows a prior visible infarct demarcation (60.7 ml, ± 74.9 ml) compared with the MonoIm (37.57 ml, ± 76.7 ml). Linear regression analysis, Bland-Altman plots and Pearson correlation coefficients show a close correlation of infarct volume in wMDIm to the fiV in SECT (r = 0.86; 95% CI 0.76-0.92), compared with MonoIm and SECT (r = 0.81; 95% CI 0.69-0.89). The agreement with SECT is substantially higher in patients with infarct volumes < 70 ml (n = 33; 66%). Coefficients were smaller with r = 0.59 (95% CI 0.31; 0.78) for MonoIm and SECT compared with r = 0.77 (95% CI 0.57; 0.88) for wMDIm and SECT. At admission, the mean NIHSS score and mRS were 17.02 (± 4.7) and 4.9 (± 0.2). mRS ≤ 2 was achieved in 56% at 90 days with a mean mRS of 2.5 (± 0.8) at discharge. CONCLUSION: Material decomposition allows earlier visibility of the final infarct volume. This promises an earlier evaluation of the dimension and severity of infarction and may lead to faster initiation of secondary stroke prophylaxis.

Impact of energy turnover on fat balance in healthy young men during energy balance, energetic restriction and overfeeding.

Body weight control is thought to be improved when physical activity and energy intake are both high (high energy turnover (ET)). The aim of the present study was to investigate the short-term impact of ET on fat balance during zero energy balance (EB), energetic restriction (ER) and overfeeding (OF). In a randomised crossover study, nine healthy men (BMI: 23·0 (SD 2·1) kg/m2, 26·6 (SD 3·5) years) passed 3 × 3 d in a metabolic chamber: three levels of ET (low, medium and high; physical activity level = 1·3-1·4, 1·5-1·6 and 1·7-1·8) were performed at zero EB, ER and OF (100, 75 and 125 % of individual energy requirement). Different levels of ET were obtained by walking (4 km/h) on a treadmill (0, 165 and 330 min). Twenty-four-hour macronutrient oxidation and relative macronutrient balance (oxidation relative to intake) was calculated, and NEFA, 24-h insulin and catecholamine secretion were analysed as determinants of fat oxidation. During EB and OF, 24-h fat oxidation increased with higher ET. This resulted in a higher relative fat balance at medium ET (EB: +17 %, OF: +14 %) and high ET (EB: +23 %, OF: +17 %) compared with low ET (all P < 0·05). In contrast, ER led to a stimulation of 24-h fat oxidation irrespective of ET (no differences in relative fat balance between ET levels, P > 0·05). In conclusion, under highly controlled conditions, a higher ET improved relative fat balance in young healthy men during OF and EB compared with a sedentary state.

Multi-arm trials with multiple primary endpoints and missing values.

We present an extension of multiple contrast tests for multiple endpoints to the case of missing values. The endpoints are assumed to be normally distributed and correlated and to have equal covariance matrices for the different treatments. Different multivariate t distributions will be applied, differing in endpoint-specific degrees of freedom. In contrast to competing methods, the familywise error type I is maintained in the strong sense in an admissible range, and the problem of different marginal errors type I is avoided. The information of all observations is exploited, thereby enabling a gain in power compared with a complete case analysis.

Deciphering the complex nature of bolting time regulation in Beta vulgaris.

KEY MESSAGE: Only few genetic loci are sufficient to increase the variation of bolting time in Beta vulgaris dramatically, regarding vernalization requirement, seasonal bolting time and reproduction type. Beta species show a wide variation of bolting time regarding the year of first reproduction, seasonal bolting time and the number of reproduction cycles. To elucidate the genetics of bolting time control, we used three F3 mapping populations that were produced by crossing a semelparous, annual sugar beet with iteroparous, vernalization-requiring wild beet genotypes. The semelparous plants died after reproduction, whereas iteroparous plants reproduced at least twice. All populations segregated for vernalization requirement, seasonal bolting time and the number of reproduction cycles. We found that vernalization requirement co-segregated with the bolting locus B on chromosome 2 and was inherited independently from semel- or iteroparous reproduction. Furthermore, we found that seasonal bolting time is a highly heritable trait (h 2 > 0.84), which is primarily controlled by two major QTL located on chromosome 4 and 9. Late bolting alleles of both loci act in a partially recessive manner and were identified in both iteroparous pollinators. We observed an additive interaction of both loci for bolting delay. The QTL region on chromosome 4 encompasses the floral promoter gene BvFT2, whereas the QTL on chromosome 9 co-localizes with the BR 1 locus, which controls post-winter bolting resistance. Our findings are applicable for marker-assisted sugar beet breeding regarding early bolting to accelerate generation cycles and late bolting to develop bolting-resistant spring and winter beets. Unexpectedly, one population segregated also for dwarf growth that was found to be controlled by a single locus on chromosome 9.

Effect of dietary quebracho tannin extract on milk fatty acid composition in cows.

The aim of this study was to examine the capacity of quebracho tannin extract (QTE) to modulate the fatty acid (FA) profile in the milk fat of cows. Fifty Holstein cows yielding 33.2 ± 8.2 kg/d of milk were divided into 2 groups. The cows were fed a basal diet with a forage-concentrate ratio of 66:34 on a dry matter (DM) basis. Diets tested were control (CON, basal diet without QTE) and basal diet plus 15 or 30 g of QTE/kg of DM (QTE15 and QTE30, respectively). Two treatments could be tested simultaneously and were arranged along 6 periods. The milk FA profile was characterized by increments in the proportion of linoleic (LA) and α-linolenic acid (α-LNA) (QTE15 = 10 and 6.1%; QTE30 = 28 and 25%, respectively) compared to CON, which might indicate reduced ruminal biohydrogenation (BH) of both dietary LA and α-LNA. Vaccenic acid (VA) in the milk fat was reduced (QTE15 8.9% and QTE30 12%) compared to CON, which may be linked to inhibited BH of LA and α-LNA. Rumenic acid (RA), a conjugated LA (cis-9,trans-11 conjugated linoleic acid) and an important human health promoter, was unfortunately decreased (QTE15 8.3% and QTE30 16%) in the milk compared with CON, probably because of inhibited ruminal BH of LA. However, reduced RA in the milk was probably due to reduced availability of VA produced in the rumen and the consequently low VA available to be desaturated to RA in the mammary gland by Δ9-desaturase. The proportions of total polyunsaturated FA were increased with QTE15 and QTE30 by 4.7 and 15% compared to CON, respectively, and the long-chain FA proportions were also increased (QTE15 2.0% and QTE30 8.2%). Moreover, myristic and palmitic acid were reduced by QTE30 (9.6 and 3.3%, respectively) compared to CON, which also contributed to increasing the nutritional quality of milk because they are recognized to increase high-density lipoprotein in humans. Branched-chain FA in milk was reduced with QTE treatments, which indicates inhibited ruminal BH and microbial activity. In general, our findings suggest that dietary QTE have the potential to modulate FA profile of milk fat, and this effect is dosage dependent. Because QTE influenced the FA profile of milk fat both positively and negatively, further research is needed before concluding that QTE may improve the nutritional quality of cow milk fat in human diets.

Effect of dietary Quebracho tannin extract on feed intake, digestibility, excretion of urinary purine derivatives and milk production in dairy cows.

The aim of this study was to evaluate the effects of dietary Quebracho tannin extract (QTE) on feed intake, apparent total tract digestibility (ATTD), excretion of urinary purine derivatives (PD) and milk composition and yield in dairy cows. Fifty Holstein cows were divided into two groups. To reach a similar performance of both groups, cows were divided according to their milk yield, body weight, days in milk and number of lactations at the start of the experiment averaging 33.2 ± 8.2 kg/d, 637 ± 58 kg, 114 ± 73 d and 2.3 ± 1.6 lactations, respectively. The cows were fed a basal diet as total mixed ration containing on dry matter (DM) basis 34% grass silage, 32% maize silage and 34% concentrate feeds. Three dietary treatments were tested, the control (CON, basal diet without QTE), QTE15 (basal diet with QTE at 15 g/kg DM) and QTE30 (basal diet with QTE at 30 g/kg DM). Two treatments were arranged along six periods each 21 d (13 d adaptation phase and 8 d sampling phase). The ATTD of DM and organic matter were reduced only in Diet QTE30, whereas both QTE treatments reduced ATTD of fibre and nitrogen (N), indicating that QTE impaired rumen fermentation. Nevertheless, feed intake was unaffected by QTE. In Diet CON, urinary N excretion accounted for 29.8% of N intake and decreased in treatments QTE15 and QTE30 to 27.5% and 17.9%, respectively. Daily faecal N excretion increased in treatments CON, QTE15 and QTE30 from 211 to 237 and 273 g/d, respectively, which amounted to 39.0%, 42.4% and 51.7% of the N intake, respectively. Hence, QTE shifted N excretion from urine to faeces, whereas the proportion of ingested N appearing in milk was not affected by QTE (average 30.7% of N intake). Daily PD excretion as indicator for microbial crude protein (CP) flow at the duodenum decreased in treatment QTE30 compared with Diet CON from 413 to 280 mmol/d. The ratios of total PD to creatinine suggest that urinary PD excretion was already lower when feeding Diet QTE15. While there was no effect of Diet QTE15, treatment QTE30 reduced milk yield, milk fat and protein. Both QTE treatments reduced milk urea concentration, which suggest that ruminal degradation of dietary CP was reduced. In summary, adding QTE at dosages of 15 and 30 g/kg DM to diets of lactating dairy cows to improve feed and protein use efficiency is not recommended.

High-Resolution Disease Phenotyping Reveals Distinct Resistance Mechanisms of Tomato Crop Wild Relatives against Sclerotinia sclerotiorum.

Besides the well-understood qualitative disease resistance, plants possess a more complex quantitative form of resistance: quantitative disease resistance (QDR). QDR is commonly defined as a partial but more durable form of resistance and, therefore, might display a valuable target for resistance breeding. The characterization of QDR phenotypes, especially of wild crop relatives, displays a bottleneck in deciphering QDR's genomic and regulatory background. Moreover, the relationship between QDR parameters, such as infection frequency, lag-phase duration, and lesion growth rate, remains elusive. High hurdles for applying modern phenotyping technology, such as the low availability of phenotyping facilities or complex data analysis, further dampen progress in understanding QDR. Here, we applied a low-cost (<1.000 €) phenotyping system to measure lesion growth dynamics of wild tomato species (e.g., Solanum pennellii or Solanum pimpinellifolium). We provide insight into QDR diversity of wild populations and derive specific QDR mechanisms and their cross-talk. We show how temporally continuous observations are required to dissect end-point severity into functional resistance mechanisms. The results of our study show how QDR can be maintained by facilitating different defense mechanisms during host-parasite interaction and that the capacity of the QDR toolbox highly depends on the host's genetic context. We anticipate that the present findings display a valuable resource for more targeted functional characterization of the processes involved in QDR. Moreover, we show how modest phenotyping technology can be leveraged to help answer highly relevant biological questions.

Relationship between animal-based on-farm indicators and meat inspection data in pigs.

BACKGROUND: This study aimed to validate slaughterhouse indicators collected during meat inspection as an alternative to on-farm animal welfare indicators. For this purpose, the assessments of twelve on-farm and seven slaughterhouse indicators of 628 pigs from three different farms were combined into three indices, differentiated between on-farm and slaughterhouse: (1) limb health, (2) other organ health, and (3) respiratory health. At first, an assessment at animal-level using agreement parameters was carried out to ascertain whether the same welfare or health issues were identified on-farm and at slaughterhouse, taking the production period (farrowing, rearing and fattening period) and the last weeks before slaughtering into account. Second, the connection of slaughterhouse findings on the individual on-farm health indices was examined using logistic regressions, to determine whether certain welfare issues can be better monitored using slaughterhouse indicators. RESULTS: Acceptable agreement was determined using the Prevalence-Adjusted Bias-Adjusted Kappa (PABAK) for the farrowing and fattening period, but not for the rearing period. A more detailed analysis of the weeks before slaughter shows that there is still a poor agreement 8 weeks before slaughter and an acceptable agreement 4 weeks before slaughter. This indicated the slaughterhouse indicators pneumonia, pleuritis and pericarditis as possible estimators of fever and deviant behavior on-farm and the slaughterhouse indicators bursitis and joint inflammations as possible estimators of lameness. In the second part of the analysis, the connection of slaughterhouse findings on the individual on-farm health indices was investigated; a significant influence of the farm on the limb and respiratory indices and no significant influence of the slaughterhouse findings could be determined, provided that all weekly assessments during the lifetime of the pigs have been taken into account. However, an influence of the slaughterhouse findings on the respiratory index and on the other organ index could be determined if only the weekly assessments four and eight weeks before slaughter, respectively, were taken into account. CONCLUSIONS: In general, the possible suitable indicators detected by the PABAK, could replace some health-related indicators but a complete substitution of on-farm welfare assessment is not possible. In addition, the traceability over time must be investigated further.

SDA coefficient is temperature dependent in rainbow trout (Oncorhynchus mykiss, Walbaum 1792) in a practical approach using group respirometry.

Rising global temperatures have raised the need for detailed knowledge of the effects of rising temperatures on the physiology of animals used in aquaculture. Here we used a multifactorial bioenergetic approach using groups of rainbow trout (Oncorhynchus mykiss) with an average single fish weight of 183.75 g ± 0.65 g to investigate the interactions of feeding and temperature with key metabolic variables. We used a recirculating aquaculture respirometry system (RARS) to test three ration sizes (0.65; 0.975; 1.3% of live body weight (BW)) over a range of three consecutive temperatures (14; 17; 20 °C). The fish were fed once per day for 6 days at each temperature and subsequently starved for 5 days to return to standard metabolic rate (SMR). This study aimed to answer the highly discussed topic of the temperature dependency of key metabolic specific dynamic action (SDA)-variables SDAcoef and SDAdur. We were able to provide evidence, that in rainbow trout the SDAcoef is highly dependent on the environmental temperature in the first ever approach to assess these variables in a group respirometer with this species. We compared the results of this study with a sophisticated bioenergetic model by Elliot and Hurley (2002) and thereby provide evidence for the practicability of group respirometry as a method to assess bioenergetic data under culture like conditions.

UVB pretreatment of ß-lactoglobulin affects the temperature-induced formation of functional amyloid-like aggregates and promotes oxidative degradation.

Unfolding in combination with or without acid hydrolysis is crucial for the formation of functional amyloid (fibrillar) or amyloid-like (worm-like) ß-lactoglobulin (BLG) aggregates, which can be induced through temperature treatment for several hours at pH 2-4. A preceding conformational destabilization of BLG might affect its aggregation. We investigated ultraviolet (UV) B radiation as conformational perturbing treatment to facilitate temperature-induced protein aggregation. 2-h UVB pretreated BLG (UV-BLG) exhibited an accelerated worm-like aggregation at pH 3.5, while at pH 2 the formation of fibrils was decelerated. The UV-induced conformational destabilization lowered the thermal stability and thus facilitates unfolding during thermal treatment. Thereby, the formation of covalent and non-covalent intermolecular interactions was favored, which promoted assembly of intact proteins resulting in worm-like aggregates. The oxidative degradation of UV-BLG was suggested to alter fibrillation-prone protein regions and thereby impede peptide assembly.

Modification and oxidative degradation of ß-lactoglobulin by UVB irradiation.

Ultraviolet (UV) B irradiation induces protein modification, especially the conformational rearrangement of proteins, and is therefore promising as a non-thermal and non-chemical functionalization technique. Nevertheless, UVB irradiation introduces radicals and oxidizes side chains resulting in the loss of food quality. Thus, assessing the UVB irradiation-based functionalization of ß-lactoglobulin (BLG) versus its oxidative degradation is of interest. UVB irradiation of up to 8 h was successfully applied to loosen the rigid folding of BLG and increase its flexibility. Thereby, the cysteine at position 121 and hydrophobic regions became surface-exposed as indicated by the increase in accessible thiol groups and increased surface hydrophobicity. Furthermore, we demonstrated the cleavage of the "outer" disulfide bond C66-C160 by LC-MS/MS after tryptic digestion of BLG. The 2-h-irradiated BLG showed adequate conformational rearrangement for protein functionalization while being minimally oxidized.

Dietary carbohydrates induce a higher SDA than lipids in rainbow trout (Oncorhynchus mykiss) based on environmental temperature.

As global temperatures increase so do the needs to investigate how the energy metabolism of fish responds to a broad range of thermal condition. Limited resources make it additionally important to use them sustainably in the feeds for aquaculture. Here we investigated the use of three different carbohydrate to lipid ratios (1:1; 1: 0.6; 1.4: 1 as non-protein energy substrates (NPES) in diets for rainbow trout (Oncorhynchus mykiss) under five different thermal regimes (12; 14; 16; 18; 20 °C) in a bioenergetic approach using a group respirometer. The results showed that the diet with carbohydrate as the main NPES resulted in a quadratic relationship of the specific dynamic action (SDA) values to temperature while diets with lipid as main NPES or a balanced ratio did not show such a response. SDA values in the diet with carbohydrate as the main NPES were significantly higher at temperatures around the optimum (15-17 °C) than the diets with lipid as the main NPES or with a balanced carbohydrate to lipid ratio. The retained energy (RE) was highly dependent on the standard metabolic rate (SMR) and SDA values did not carry over onto them. The protein utilization for energy combustion was significantly lower at 12 °C in the diet with carbohydrate as the main NPES than in the diet with lipid as the main NPES thus indicating that carbohydrates hold a relevant nutritional value especially at lower temperatures.

Analysis of the adiponectin paradox in healthy older people.

BACKGROUND: It remains unknown why adiponectin levels are associated with poor physical functioning, skeletal muscle mass and increased mortality in older populations. METHODS: In 190 healthy adults (59-86 years, BMI 17-37 kg/m2 , 56.8% female), whole body skeletal muscle mass (normalized by height, SMI, kg/m2 ), muscle and liver fat were determined by magnetic resonance imaging. Bone mineral content (BMC) and density (BMD) were assessed by dual X-ray absorptiometry (n = 135). Levels of insulin-like growth factor 1 (IGF-1), insulin, inflammation markers, leptin and fibroblast growth factor 21 were measured as potential determinants of the relationship between adiponectin and body composition. RESULTS: Higher adiponectin levels were associated with a lower SMI (r = -0.23, P < 0.01), BMC (r = -0.17, P < 0.05) and liver fat (r = -0.20, P < 0.05) in the total population and with higher muscle fat in women (r = 0.27, P < 0.01). By contrast, IGF-1 showed positive correlations with SMI (r = 0.33), BMD (r = 0.37) and BMC (r = 0.33) (all P < 0.01) and a negative correlation with muscle fat (r = -0.17, P < 0.05). IGF-1 was negatively associated with age (r = -0.21, P < 0.01) and with adiponectin (r = -0.15, P < 0.05). Stepwise regression analyses revealed that IGF-1, insulin and leptin explained 18% of the variance in SMI, and IGF-1, leptin and age explained 16% of the variance in BMC, whereas adiponectin did not contribute to these models. CONCLUSIONS: Associations between higher adiponectin levels and lower muscle or bone mass in healthy older adults may be explained by a decrease in IGF-1 with increasing adiponectin levels.

Determinants of bone mass in older adults with normal- and overweight derived from the crosstalk with muscle and adipose tissue.

Lower bone mass in older adults may be mediated by the endocrine crosstalk between muscle, adipose tissue and bone. In 150 community-dwelling adults (59-86 years, BMI 17-37 kg/m2; 58.7% female), skeletal muscle mass index, adipose tissue and fat mass index (FMI) were determined. Levels of myokines, adipokines, osteokines, inflammation markers and insulin were measured as potential determinants of bone mineral content (BMC) and density (BMD). FMI was negatively associated with BMC and BMD after adjustment for mechanical loading effects of body weight (r-values between -0.37 and -0.71, all p < 0.05). Higher FMI was associated with higher leptin levels in both sexes, with higher hsCRP in women and with lower adiponectin levels in men. In addition to weight and FMI, sclerostin, osteocalcin, leptin × sex and adiponectin were independent predictors of BMC in a stepwise multiple regression analysis. Muscle mass, but not myokines, showed positive correlations with bone parameters that were weakened after adjusting for body weight (r-values between 0.27 and 0.58, all p < 0.01). Whereas the anabolic effect of muscle mass on bone in older adults may be partly explained by mechanical loading, the adverse effect of obesity on bone is possibly mediated by low-grade inflammation, higher leptin and lower adiponectin levels.

Multiple comparisons to both a negative and a positive control.

This paper addresses multiple comparisons in the presence of both a negative and a positive control. The methodology of the three-arm trial is extended to the case of many experimental treatment arms or different doses of a compound. In contrast to the classic three-arm trial, the focus is on the family-wise error type I. Normally distributed data with either homogeneous or heterogeneous group variances are considered. Explicit criteria for an optimal allocation are proposed. Depending on the pattern of heterogeneity, remarkably unbalanced designs are power-optimal. As an example, the method will be applied to a toxicological experiment.