Three-Dimensional Imaging of Bioinspired Lipidic Mesophases Using Multicolored Light-Emitting Carbon Nanodots.

Recent progress in the design of carbon nanostructures exhibiting strong multiphoton-excited emission opens new pathways to explore the self-organization of lipids found in living organisms. Phospholipid-based lyotropic myelin figures (MFs) are promising materials as simplified models of biomembranes due to their structural resemblance to a multilamellar sheath insulating the axon. This study demonstrates the possibility of selective labeling of MFs by strongly emitting multicolor phloroglucinol-derived carbon nanodots (PG CNDs). Such dopants are efficiently excited by visible and near-infrared light; therefore, one- and two-photon fluorescence microscopies are incorporated to gain 3D insights into the MFs. Combining nondestructive fluorescence microscopy and spectroscopy techniques along with polarized light microscopy gives details on the stability and morphology of lipidic mesophases. Our findings suggest that PG CNDs can be a viable and simple alternative to conventional fluorescent lipid stains to image biologically relevant phospholipid-based structures.

Strongly Emitting Folic Acid-Derived Carbon Nanodots for One- and Two-Photon Imaging of Lyotropic Myelin Figures.

Non-invasive imaging of morphological changes in biologically relevant lipidic mesophases is essential for the understanding of membrane-mediated processes. However, its methodological aspects need to be further explored, with particular attention paid to the design of new excellent fluorescent probes. Here, we have demonstrated that bright and biocompatible folic acid-derived carbon nanodots (FA CNDs) may be successfully applied as fluorescent markers in one- and two-photon imaging of bioinspired myelin figures (MFs). Structural and optical properties of these new FA CNDs were first extensively characterized; they revealed remarkable fluorescence performance in linear and non-linear excitation regimes, justifying further applications. Then, confocal fluorescence microscopy and two-photon excited fluorescence microscopy were used to investigate a three-dimensional distribution of FA CNDs within the phospholipid-based MFs. Our results showed that FA CNDs are effective markers for imaging various forms and parts of multilamellar microstructures.

Including genotypic information in genetic evaluations increases the accuracy of sheep breeding values.

The impact of inclusion of genome-wide genotypes into breeding value predictions for UK Texel sheep is addressed in this article. The main aim was to investigate the level of change in the accuracy values for EBVs when information from animal genotypes is incorporated into the genetic evaluations. New genetic parameters for a range of lamb growth, carcass composition and health traits are described and applied in the estimation of conventional breeding values (EBVs) for almost 822,000 animals as well as genomic breeding values (gEBVs) after adding 10,143 genotypes. Principal component analyses showed that there are no major distinct groups; hence, the population is mainly homogenous and genetically well-linked. Results suggested that the highest change in accuracy was observed for the animals that are not phenotyped but have good links to the reference population. This was seen especially for the lowly heritable health traits thereby proving that the use of genotypes in breeding values estimation may accelerate the genetic gain by producing more accurate values especially for young, un-phenotyped animals.

Non-toxic Polymeric Dots with the Strong Protein-Driven Enhancement of One- and Two-Photon Excited Emission for Sensitive and Non-destructive Albumin Sensing.

The need for efficient probing, sensing, and control of the bioactivity of biomolecules (e.g., albumins) has led to the engineering of new fluorescent albumins' markers fulfilling very specific chemical, physical, and biological requirements. Here, we explore acetone-derived polymer dots (PDs) as promising candidates for albumin probes, with special attention paid to their cytocompatibility, two-photon absorption properties, and strong ability to non-destructively interact with serum albumins. The PDs show no cytotoxicity and exhibit high photostability. Their pronounced green fluorescence is observed upon both one-photon excitation (OPE) and two-photon excitation (TPE). Our studies show that both OPE and TPE emission responses of PDs are proteinaceous environment-sensitive. The proteins appear to constitute a matrix for the dispersion of fluorescent PDs, limiting both their aggregation and interactions with the aqueous environment. It results in a large enhancement of PD fluorescence. Meanwhile, the PDs do not interfere with the secondary protein structures of albumins, nor do they induce their aggregation, enabling the PD candidates to be good nanomarkers for non-destructive probing and sensing of albumins.

The Effect of Rumination Time on Milk Performance and Methane Emission of Dairy Cows Fed Partial Mixed Ration Based on Maize Silage.

The objective of this study was to determine the effect of the rumination time on milk yield and composition as well as methane emission during lactation in high-yielding dairy cows fed a partial mixed ration based on maize silage without pasture access. A total of 365 high-yielding Polish Holstein-Friesian multiparous dairy cows were included in the study covering 24 to 304 days of lactation. Methane emission, rumination time, and milk production traits were observed for the period of 12 months. Next, the data from the cows were assigned to three groups based on daily rumination time: low rumination up to 412 min/day (up to 25th rumination percentile), medium rumination from 412 to 527 min/day (between the 25th and 75th percentile), and high rumination above 527 min/day (from the 75th percentile). Rumination time had no effect on milk yield, energy-corrected milk yield, or fat and protein-corrected milk yield. High rumination time had an effect on lower fat concentration in milk compared with the medium and low rumination groups. The highest daily CH4 production was noted in low rumination cows, which emitted 1.8% more CH4 than medium rumination cows and 4.2% more than high rumination cows. Rumination time affected daily methane production per kg of milk. Cows from the high rumination group produced 2.9% less CH4 per milk unit compared to medium rumination cows and 4.6% in comparison to low rumination cows. Similar observations were noted for daily CH4 production per ECM unit. In conclusion, a longer rumination time is connected with lower methane emission as well as lower methane production per milk unit in high-yielding dairy cows fed a maize silage-based partial mixed ration without pasture access.

Acetone-derived luminescent polymer dots: a facile and low-cost synthesis leads to remarkable photophysical properties.

Carbon-based dots have been attracting much attention as potentially superior alternatives to more conventional semiconductor nanoparticles, due to their fascinating optical properties, chemical and photochemical stability, unique environmental-friendliness, and the versatility of fabrication routes. Many commercial materials and organic compounds have been considered so far as carbon precursors but in many cases the fabrication required high-temperature conditions or led to inhomogeneous final products. Here we report on a simple low-cost synthesis of non-conjugated carbon-rich polymer dots (PDs) that uses acetone as carbon precursor. Both hydrophilic and hydrophobic fractions of PDs were obtained, with the respective average diameters of 2-4 nm and ca. 6 nm. The as-obtained PDs reveal greenish-blue photoluminescence (PL) and high quantum yields (∼5-7%) and complex kinetics of the decays with the average lifetime of ∼3.5 ns. Such luminescent acetone-derived PDs may find application in several fields, including sensing and bioimaging.

Genetic analysis of novel phenotypes for farm animal resilience to weather variability.

BACKGROUND: Climate change is expected to have a negative impact on food availability. While most efforts have been directed to reducing greenhouse gas emissions, complementary strategies are necessary to control the detrimental effects of climate change on farm animal performance. The objective of this study was to develop novel animal resilience phenotypes using reaction norm slopes, and examine their genetic and genomic parameters. A closely monitored dairy goat population was used for this purpose. RESULTS: Individual animals differed in their response to changing atmospheric temperature and a temperature-humidity index. Significant genetic variance and heritability estimates were derived for these animal resilience phenotypes. Furthermore, some resilience traits had a significant unfavourable genetic correlation with animal performance. Genome-wide association analyses identified several candidate genes related to animal resilience to environment change. CONCLUSIONS: Heritable variation exists among dairy goats in their production response to fluctuating weather variables. Results may inform future breeding programmes aimed to ensure efficient animal performance under changing climatic conditions.

Genome-wide association study of conformation and milk yield in mixed-breed dairy goats.

Identification of genetic markers that affect economically important traits is of high value from a biological point of view, enabling the targeting of candidate genes and providing practical benefits for the industry such as wide-scale genomic selection. This study is one of the first to investigate the genetic background of economically important traits in dairy goats using the caprine 50K single nucleotide polymorphism (SNP) chip. The aim of the project was to perform a genome-wide association study for milk yield and conformation of udder, teat, and feet and legs. A total of 137,235 milk yield records on 4,563 goats each scored for 10 conformation traits were available. Out of these, 2,381 goats were genotyped with the Illumina Caprine 50K BeadChip (Illumina Inc., San Diego, CA). A range of pseudo-phenotypes were used including deregressed breeding values and pseudo-estimated breeding values. Genome-wide association studies were performed using the multi-locus mixed model (MLMM) algorithm implemented in SNP & Variation Suite v7.7.8 (Golden Helix Inc., Bozeman, MT). A genome-wise significant [-log10(P-value) > 5.95] SNP for milk yield was identified on chromosome 19, with additional chromosome-wise significant (-log10(P-value) > 4.46] SNP on chromosomes 4, 8, 14, and 29. Three genome-wise significant SNP for conformation of udder attachment, udder depth, and front legs were identified on chromosome 19, and chromosome-wise SNP were found on chromosomes 4, 5, 6, 10, 11, 12, 13, 14, 15, 16, 17, 18, 21, 23, and 27. The proportion of variance explained by the significant SNP was between 0.4 and 7.0% for milk yield and between 0.1 and 13.8% for conformation traits. This study is the first attempt to identify SNP associated with milk yield and conformation in dairy goats. Two genome-wise significant SNP for milk yield and 3 SNP for conformation of udder attachment, udder depth, and front legs were found. Our results suggest that conformation traits have a polygenic background because, for most of them, we did not identify any quantitative trait loci with major effect.

Hypoglossal nerve palsy in the course of dissection of the internal carotid arteries - Case reports.

Internal carotid artery dissection (ICAD) has become an increasingly recognized cause of cerebrovascular accidents in young and middle-aged patients. We report 2 cases of hypoglossal nerve palsy in the course of dissection of the internal carotid arteries. The first patient was admitted to the Department of Neurology due to swallowing difficulty, speech articulation disorders and numbness of the right half of the tongue for 4 weeks. Extracranial vessel ultrasound (US) and transcranial colour Doppler (TCD) visualized thrombus causing occlusion of the right internal carotid artery (RICA). Angio-CT revealed a compression on right XII nerve and a dissection of the RICA. The second patient was referred to the Department of Neurology due to articulation disorders and swallowing difficulties. On admission, neurological examination revealed tongue deviation towards the right side with evidence of atrophy of the right half of the tongue, deviation of the uvula to the right side, absence of palatal and pharyngeal reflexes, rhinolalia and dysphagia. Vessel imaging was taken using angio-MR showing mural thrombus of the RICA. CONCLUSION: The diagnosis of spontaneous non-traumatic dissection of the carotid arteries is a major challenge for clinicians. ICAD must be considered for young and middle-aged patients when severe headache is preceded by the co-existence of focal neurological symptoms. The probability of ICAD increases in the presence of predisposing diseases. The final diagnosis is based on imaging studies: color duplex ultrasound, CT angiography or MR angiography.

Changes in Serum Ceruloplasmin Levels Based on Immunomodulatory Treatments and Melatonin Supplementation in Multiple Sclerosis Patients.

BACKGROUND The cause of multiple sclerosis (MS) is currently unknown, but it is thought that oxidative damage and iron metabolism mechanisms are involved. The aim of this study was to examine ceruloplasmin concentration in MS patients based on various immunomodifying therapies and to test the effect of antioxidative melatonin on ceruloplasmin levels. MATERIAL AND METHODS This prospective study included 102 MS patients and 15 healthy controls. Patients were divided into groups according to different immunomodifying therapies: interferons beta 1a, interferons beta 1b, glatiramer acetate, mitoxantrone, and immunomodifying pre-treatment (A, B, G, Mx, and P groups, respectively), and the relapse R group. MS patients were supplemented with melatonin for 3 months. Serum ceruloplasmin concentrations, EDSS, brain MRI, serum C-reactive protein level, and white blood cell count were examined. RESULTS The results indicated significantly increased levels of ceruloplasmin in MS patients. No differences in ceruloplasmin concentrations between the relapse group and controls were observed. In A and G groups, ceruloplasmin levels before and after melatonin were similar to levels in controls. In group B, ceruloplasmin concentration was significantly higher vs. control and relapse groups. After melatonin administration in group B, ceruloplasmin levels decreased. Ceruloplasmin concentrations in the Mx group were significantly higher compared to controls. CONCLUSIONS We found for the first time that ceruloplasmin concentration in MS patients varies depending on different immunomodulatory treatment and decrease after 3 months of melatonin administration. Ceruloplasmin could be a valuable serum marker for the chronic demyelinating process participating in oxidative stress mechanisms, as well as a neurodegenerative marker, but not a marker of acute-phase MS.

Genome-wide association study of footrot in Texel sheep.

BACKGROUND: This is the first study based on a genome-wide association approach that investigates the links between ovine footrot scores and molecular polymorphisms in Texel sheep using the ovine 50 K SNP array (42 883 SNPs (single nucleotide polymorphisms) after quality control). Our aim was to identify molecular predictors of footrot resistance. METHODS: This study used data from animals selected from a footrot-phenotyped Texel sheep population of 2229 sheep with an average of 1.60 scoring records per animal. From these, a subset of 336 animals with extreme trait values for footrot was selected for genotyping based on their phenotypic records. De-regressed estimated breeding values (EBV) for footrot were used as pseudo-phenotypes in the genome-wide association analysis. RESULTS: Seven SNPs were significant on a chromosome-wise level but the association analysis did not reveal any genome-wise significant SNPs associated with footrot. Based on the current state of knowledge of the ovine genome, it is difficult to clearly link the function of the genes that contain these significant SNPs with a potential role in resistance/susceptibility to footrot. Linkage disequilibrium (LD) was analysed as one of the factors that influence the power of detecting QTL (quantitative trait loci). A mean LD of 0.20 (r(2) at a distance of 50 kb between two SNPs) in the population analysed was estimated. LD declined from 0.15 to 0.07 and to 0.04 at distances between two SNPs of 100, 1000 and 2000 kb, respectively. CONCLUSIONS: Based on a relatively small number of genotyped animals, this study is a first step to search for genomic regions that are involved in resistance to footrot using the ovine 50 K SNP array. Seven SNPs were found to be significant on a chromosome-wise level. No major genome-wise significant QTL were identified.

Melatonin acts as antioxidant and improves sleep in MS patients.

The relationship between the prevalence of multiple sclerosis (MS) and sunlight's ultraviolet radiation was proved. Oxidative stress plays a role in the pathogenic traits of MS. Melatonin possesses antioxidative properties and regulates circadian rhythms. Sleep disturbances in MS patients are common and contribute to daytime fatigue. The aim of study was to evaluate 5 mg daily melatonin supplementation over 90 days on serum total oxidant status (TOS), total antioxidant capacity (TAC) and its influence on sleep quality and depression level of MS patients. A case-control prospective study was performed on 102 MS patients and 20 controls matched for age and sex. The Kurtzke's Expanded Disability Status Scale, magnetic resonance imaging examinations, Athens Insomnia Scale (AIS), Beck Depression Inventory questionnaires were completed. Serum TOS and TAC levels were measured. We observed higher serum levels of TOS in all MS groups, while after melatonin treatment the TOS levels significantly decreased. The TAC level was significantly lower only in mitoxantrone-treated group and it increased after melatonin supplementation. A strong positive correlation between T1Gd(+) number lesions and TAC level in interferon-beta-1A group was observed. AIS group mean score above 6 defining insomnia were observed in interferon-beta-1B-group, glatiramer acetate-group and mitoxantrone-group: 6.62 ± 2.88, 8.45 ± 2.07, 11.1 ± 3.25, respectively. After melatonin treatment the AIS mean scores decrease in glatiramer acetate-group and mitoxantrone-group achieving 5.25 ± 1.14 and 7.08 ± 2.39, respectively (p < 0.05). Finding from our study suggest that melatonin can act as an antioxidant and improves reduced sleep quality in MS patients.

Mapping quantitative trait loci affecting some carcass and meat traits in duck (Anas platyrhynchos).

Contrary to chicken and livestock mammals, duck genome has not been explored much. Nowadays a relatively small number of reports on molecular variability and mapping of loci in Peking ducks has been published. Therefore, the objective of this study was to detect single loci affecting body weight, carcass and meat traits in Peking ducks (Anas platyrhynchos). The study was based on an F2 cross between two parental lines A-55 and GL-30. Phenotypes of 387 birds from generation F2 including carcass and meat quality traits were collected. Linkage map, of the linkage group CAU1, consisting of 29 microsatellite markers was constructed. One highly significant (p < 0.01) QTL for skin weight was identified at the beginning (0 cM) of the linkage group CAU1. Two significant QTL (p < 0.05) were found for fatness of leg muscle and leg muscle electrical conductivity - 24 hours post slaughtering (EC24h) at 178 and 100 cM, respectively. Additionally one suggestive QTL for odour of leg muscle was identified at 265 cM.

Oxidative modification of serum proteins in multiple sclerosis.

Multiple sclerosis (MS) has been demonstrated to involve oxidative stress and augmented glycoxidation. In this study, several markers of protein oxidative damage and glycoxidation have been compared in 14 relapsing remittent in MS (RRMS) patients without immunomodifying treatment, 10 patients in clinical relapse, and clinically stable patient groups treated with interferon ß 1a (18) , ß 1b (19) and glatiramer acetate (GA; 6) in relation to healthy subjects (12). The glycophore content was increased in RRSM patients without treatment and in patients treated with GA. The level of advanced protein oxidation products (AOPP) was increased in RRSM patients without treatment and in patients with clinical relapse. The level of protein carbonyls was elevated in RRSM patients without treatment and in patients treated with interferon ß 1b. The levels of dityrosine level and N'-formylkynureine were elevated in RRSM patients without treatment while serum protein thiol groups were decreased in RRSM patients in clinical relapse as well as RRMS patients treated with interferon ß 1a. Several markers of protein modification showed correlation with the C-reactive protein level and white blood cell count, suggesting that oxidative protein modifications are linked to the inflammatory processes in MS. Results of this study confirm the occurrence of protein oxidative and glycoxidative damage in MS and show that spectrophotometric and fluorimetric markers of this damage, especially the AOPP level, may be useful in monitoring oxidative stress in the course of therapy of MS.

Comparison of analyses of the QTLMAS XIV common dataset. I: genomic selection.

BACKGROUND: For the XIV QTLMAS workshop, a dataset for traits with complex genetic architecture has been simulated and released for analyses by participants. One of the tasks was to estimate direct genomic values for individuals without phenotypes. The aim of this paper was to compare results of different approaches used by the participants to calculate direct genomic values for quantitative trait (QT) and binary trait (BT). RESULTS: Participants applied 26 approaches for QT and 15 approaches for BT. Accuracy for QT was between 0.26 and 0.89 for males and between 0.31 and 0.89 for females, and for BT ranged from 0.27 to 0.85. For QT, percentage of lost response to selection varied from 8% to 83%, whereas for BT the loss was between 15% and 71%. CONCLUSIONS: Bayesian model averaging methods predicted breeding values slightly better than GBLUP in a simulated data set. The methods utilizing genomic information performed better than traditional pedigree based BLUP analyses. Bivariate analyses was slightly advantageous over single trait for the same method. None of the methods estimated the non-additivity of QTL affecting the QT, which may be one of the constrains in accuracy observed in real data.

Comparison of analyses of the QTLMAS XIV common dataset. II: QTL analysis.

BACKGROUND: A quantitative and a binary trait for the 14th QTLMAS 2010 workshop were simulated under a model which combined additive inheritance, epistasis and imprinting. This paper aimed to compare results submitted by the participants of the workshop. METHODS: The results were compared according to three criteria: the success rate (ratio of mapped QTL to the total number of simulated QTL), and the error rate (ratio of false positives to the number of reported positions), and mean distance between a true mapped QTL and the nearest submitted position. RESULTS: Seven groups submitted results for the quantitative trait and five for the binary trait. Among the 37 simulated QTL 17 remained undetected. Success rate ranged from 0.05 to 0.43, error rate was between 0.00 and 0.92, and the mean distance ranged from 0.26 to 0.77 Mb. CONCLUSIONS: Our comparison shows that differences among methods used by the participants increases with the complexity of genetic architecture. It was particularly visible for the quantitative trait which was determined partly by non-additive QTL. Furthermore, an imprinted QTL with a large effect may remain undetected if the applied model tests only for Mendelian genes.

Comparison of methods for estimation of genetic covariance matrix from SNP or pedigree data utilised to predict breeding value.

BACKGROUND: The aim was to predict breeding values of non-phenotyped individuals based on a dataset prepared for the 13th QTL-MAS Workshop in Wageningen. METHODS: Genetic co-variance matrices between animals were estimated with three methods: one using pedigree information only and two based on SNP markers from the first chromosome. Quadratic regression of breeding values, estimated separately in each of the five time points, was used to predict the breeding values in the 6th time point. RESULTS: Based on the comparison (true - estimated BV) it can be concluded that SNP based methods provided better estimates (accuracy between 0.75 and 0.80) than pedigree (0.65). CONCLUSIONS: Even though only SNPs from chromosome 1 were used it was still possible to achieve fairly high accuracies. Most likely this was due to the fact that chromosome 1 contained the QTLs with the largest effects.