Interferon-gamma producing CD4+ T cells quantified by flow cytometry as early markers for Mycobacterium avium ssp. paratuberculosis infection in cattle.

Current diagnostic methods for Johne's disease in cattle allow reliable detection of infections with Mycobacterium avium ssp. paratuberculosis (MAP) not before animals are 2 years of age. Applying a flow cytometry-based approach (FCA) to quantify a MAP-specific interferon-gamma (IFN-γ) response in T cell subsets, the present study sought to monitor the kinetics of the cell-mediated immune response in experimentally infected calves. Six MAP-negative calves and six calves, orally inoculated with MAP at 10 days of age, were sampled every 4 weeks for 52 weeks post-inoculation (wpi). Peripheral blood mononuclear cells (PBMC) were stimulated with either purified protein derivatives (PPD) or whole cell sonicates derived from MAP (WCSj), M. avium ssp. avium or M. phlei for 6 days followed by labeling of intracellular IFN-γ in CD4+ and CD8+ T cells. No antigen-specific IFN-γ production was detectable in CD8+ cells throughout and the responses of CD4+ cells of MAP-infected and control calves were similar up to 12 wpi. However, the mean fluorescence intensity (MFI) for the detection of IFN-γ in CD4+ cells after WCSj antigen stimulation allowed for a differentiation of animal groups from 16 wpi onwards. This approach had a superior sensitivity (87.8%) and specificity (86.8%) to detect infected animals from 16 wpi onwards, i.e., in an early infection stage, as compared to the IFN-γ release assay (IGRA). Quantification of specific IFN-γ production at the level of individual CD4+ cells may serve, therefore, as a valuable tool to identify MAP-infected juvenile cattle.

The role of the A monovalent cation in the AVF3 perovskite series. A quantum mechanical investigation.

The relative stability of various phases of five AVF3 compounds (A = Li, Na, K, Rb and Cs) is investigated starting from the cubic (C) Pm3̄m (221) prototype structure, with five atoms (one formula unit) in the primitive cell. To the authors' knowledge, only three of these compounds have been investigated experimentally (Na, K and Rb), and they are reported as being cubic. The picture emerging from the present simulation is quite different: CsVF3 and RbVF3 are dynamically stable in the cubic structure, KVF3 is tetragonal, with space group (SG) I4/mcm (no. 140) and 10 atoms in the unit cell; note, however, that an orthorhombic Pnma (62) phase (four formula units) exists, which is not a subgroup of I4/mcm (140), and is very close in energy to the tetragonal phase. Further symmetry lowering is observed in the Na and Li compounds that are orthorhombic. The energy gain and volume reduction with respect to the cubic aristotype increase along the series K, Na and Li, and is very large for the last two compounds. Both FM and AFM solutions have been explored, and they show a very similar path along the SG modifications. The present scheme for determining the lowest energy SG is general, and can be applied to any perovskite. The B3LYP full range hybrid functional and the Hartree-Fock (HF) Hamiltonian, an all-electron Gaussian type basis set and the CRYSTAL code have been used.

How deeply are core electrons perturbed when valence electrons are spin polarized? The case study of transition metal compounds.

The ferromagnetic and antiferromagnetic wave functions of the KMnF3 perovskite have been evaluated quantum-mechanically by using an all electron approach and, for comparison, pseudopotentials on the transition metal and the fluorine ions. It is shown that the different number of α and ß electrons in the d shell of Mn perturbs the inner shells, with shifts between the α and ß eigenvalues that can be as large as 6 eV for the 3s level, and is far from negligible also for the 2s and 2p states. The valence electrons of F are polarized by the majority spin electrons of Mn, and in turn, spin polarize their 1s electrons. When a pseudopotential is used, such a spin polarization of the core functions of Mn and F can obviously not take place. The importance of such a spin polarization can be appreciated by comparing (i) the spin density at the Mn and F nuclear position, and then the Fermi contact constant, a crucial quantity for the hyperfine coupling, and (ii) the ferromagnetic-antiferromagnetic energy difference, when obtained with an all electron or a pseudopotential scheme, and exploring how the latter varies with pressure. This difference is as large as 50% of the all electron datum, and is mainly due to the rigid treatment of the F ion core. The effect of five different functionals on the core spin polarization is documented.

The role of spin density for understanding the superexchange mechanism in transition metal ionic compounds. The case of KMF3 (M = Mn, Fe, Co, Ni, Cu) perovskites.

In many recent papers devoted to first row transition metal fluorides and oxides, not much attention is devoted to the spin density, a crucial quantity for the determination of the superexchange mechanism, and then for the ferro-antiferromagnetic energy difference. Usually, only the eigenvalues of the system are represented, in the form of band structures or, more frequently, of density of states (DOS). When discussing the orbital ordering and the Jahn-Teller effect, simple schemes with cubes and lobes are used to illustrate the shape of the d occupancy. But the eigenvectors, and the resulting spin density function, as obtained from the calculations, are rarely shown. When represented, only a fuzzy shape that recalls the d orbital shape can be observed. On the basis of these considerations, spin density maps for 5 compounds of the KMF3 (M from Mn to Cu) family have been produced, which clearly illustrate which d orbital is singly or doubly occupied. At variance with respect to the near totality of the papers devoted to these systems, we use an all electron scheme, a Gaussian type basis set, and the Hartree-Fock Hamiltonian or the B3LYP hybrid functional (the resulting maps turn out to be very similar, in the scale used for our figures). The spin density in the five cases can easily be interpreted in terms of the shape of the d orbitals as appearing in textbooks.

The CRYSTAL code, 1976-2020 and beyond, a long story.

CRYSTAL is a periodic ab initio code that uses a Gaussian-type basis set to express crystalline orbitals (i.e., Bloch functions). The use of atom-centered basis functions allows treating 3D (crystals), 2D (slabs), 1D (polymers), and 0D (molecules) systems on the same grounds. In turn, all-electron calculations are inherently permitted along with pseudopotential strategies. A variety of density functionals are implemented, including global and range-separated hybrids of various natures and, as an extreme case, Hartree-Fock (HF). The cost for HF or hybrids is only about 3-5 times higher than when using the local density approximation or the generalized gradient approximation. Symmetry is fully exploited at all steps of the calculation. Many tools are available to modify the structure as given in input and simplify the construction of complicated objects, such as slabs, nanotubes, molecules, and clusters. Many tensorial properties can be evaluated by using a single input keyword: elastic, piezoelectric, photoelastic, dielectric, first and second hyperpolarizabilities, etc. The calculation of infrared and Raman spectra is available, and the intensities are computed analytically. Automated tools are available for the generation of the relevant configurations of solid solutions and/or disordered systems. Three versions of the code exist: serial, parallel, and massive-parallel. In the second one, the most relevant matrices are duplicated on each core, whereas in the third one, the Fock matrix is distributed for diagonalization. All the relevant vectors are dynamically allocated and deallocated after use, making the code very agile. CRYSTAL can be used efficiently on high performance computing machines up to thousands of cores.

The Molpro quantum chemistry package.

Molpro is a general purpose quantum chemistry software package with a long development history. It was originally focused on accurate wavefunction calculations for small molecules but now has many additional distinctive capabilities that include, inter alia, local correlation approximations combined with explicit correlation, highly efficient implementations of single-reference correlation methods, robust and efficient multireference methods for large molecules, projection embedding, and anharmonic vibrational spectra. In addition to conventional input-file specification of calculations, Molpro calculations can now be specified and analyzed via a new graphical user interface and through a Python framework.

Monitoring of iron deficiency in calves by determination of serum ferritin in comparison with serum iron: A preliminary study.

Background: Iron deficiency can cause anemia in calves and is, therefore, of economic importance for the cattle industry. Low iron levels are commonly caused by feeding whole milk without the addition of dietary supplements and led to the most frequent cause of anemia in calves. Other reasons for the development of anemia include congenital iron deficiency, malnutrition, bleeding ulcers, or bloodsucking parasites. Aim: This study compared laboratory parameters that are commonly used to diagnose iron deficiency anemia in calves. Additionally, serum ferritin values were compared amongst calves fed different milk meals. Methods: For this purpose, blood samples from 40 calves were analyzed for different hematologic parameters as well as the content of copper, glutathione peroxidase, serum iron, and serum ferritin. Results: Eight calves showed decreased hemoglobin and hematocrit values and a significantly lower number of erythrocytes compared with non-anemic calves. Interestingly, 19 of 40 calves had a low serum iron. Considering their serum ferritin levels, only 14 calves, including six calves with both low iron and low ferritin levels, were classified as iron deficient. No direct correlation between serum ferritin and serum iron was detected. Comparing milk diets, more calves fed milk replacer showed reduced levels of ferritin compared with calves fed whole milk. Conclusion: Our data indicate that the determination of hemoglobin and serum iron is of limited suitability for the diagnosis of iron deficiency in calves. We suggest that the determination of serum ferritin should be the preferred parameter, since serum iron levels are subject to physiological fluctuation and a deficiency can be caused by inflammation or neoplastic diseases.

Black Current: Structure, Characterization, and Optoelectronic Properties of Ce3 Cl3 [MoO6 ].

The admixture of CeO2 , Ce, CeCl3 , and MoO3 with an excess of LiCl as flux in evacuated silica ampules leads to large black single crystals as well as a black microcrystalline powder of Ce3 Cl3 [MoO6 ] after tempering at 850 °C for three days. The title compound crystallizes in the hexagonal space group P63 /m (a=934.93(4), c=538.86(2) pm) with two formula units per unit cell. The crystal structure consists of rather unusual trigonal-prismatic [MoO6 ]6- units besides Ce3+ ions in a tetra-capped trigonal-prismatic coordination, formed by four Cl- and six O2- ions. The black color is related to an optical band gap of 1.35(2) eV, which was determined by diffuse reflectance spectroscopy and confirmed by theoretical calculations. The low band gap between the 4f1 state of cerium (HOMO) and the 5d0 state of molybdenum (LUMO) gave rise to the idea of electronic excitation between these two states by IR irradiation, creating a drop in the resistivity of the material, which was detected by appropriate measurements.

Barium Sulfide under Pressure: Discovery of Metastable Polymorphs and Investigation of Electronic Properties on ab Initio Level.

Barium sulfide (BaS) is an important precursor to other barium compounds with applications from ceramics and flame retardants to luminous paints and additives, and recent research shows potential technological applications in electrical and optical devices. Under normal conditions, BaS crystallizes in the NaCl type of structure, and with the increase in pressure BaS undergoes a structural phase transition to a CsCl type modification. This study presents modeling of barium sulfide under pressure with special focus on structural aspects and electronic properties. We predict metastable BaS polymorphs which have not yet been observed in the experiment or in previous calculations, and we investigated their vibrational and thermodynamical properties. Furthermore, we investigate the electronic properties of experimentally known structures as well as novel predicted modifications of BaS on ab initio level using Hartree-Fock, GGA-PBE, and the hybrid B3LYP functional. In this way, we address new possibilities of synthesizing BaS and possible band gap tuning which can have great applications in optoelectrical technologies.

Electronic and transport properties of LiCoO2.

Using first principles density functional theory (DFT), the electronic and magnetic properties as well as the Li-ion migration in LiCoO2 have been studied with a gradient corrected functional. The magnetic properties were also investigated in addition using a gradient corrected functional in combination with an on-site repulsion U and a hybrid functional. We find LiCoO2 to be non-magnetic under ambient conditions. A magnetic ground state can be obtained by a volume expansion corresponding to a negative pressure of -8 GPa due to a competition between Hund's rules favoring magnetism on the Co(3+) ions and the crystal field splitting, which suppresses magnetism at zero pressure. The barrier for lithium transport is determined to be 0.44 eV from nudged elastic band (NEB) calculations on the Li0.917CoO2 system.

Sterically active electron pairs in lead sulfide? An investigation of the electronic and vibrational properties of PbS in the transition region between the rock salt and the α-GeTe-type modifications.

Recently, we have investigated the energy landscape of PbS for many different pressures on the ab initio level by using Hartree-Fock and density functional theory to globally search for possible thermodynamically stable and metastable structures. The perhaps most fascinating observation was that besides the experimentally known modification exhibiting the rock salt structure a second minimum exists close-by on the landscape showing the low-temperature α-GeTe-type structure. In the present study, we investigate the possible reasons for the existence of this metastable modification; in particular we address the question, whether the α-GeTe-type modification might be stabilized (and conversely the rock salt modification destabilized) by steric effects of the non-bonding electron pair.

Transcription factor binding site polymorphism in the motilin gene associated with left-sided displacement of the abomasum in German Holstein cattle.

Left-sided displacement of the abomasum (LDA) is a common disease in many dairy cattle breeds. A genome-wide screen for QTL for LDA in German Holstein (GH) cows indicated motilin (MLN) as a candidate gene on bovine chromosome 23. Genomic DNA sequence analysis of MLN revealed a total of 32 polymorphisms. All informative polymorphisms used for association analyses in a random sample of 1,136 GH cows confirmed MLN as a candidate for LDA. A single nucleotide polymorphism (FN298674:g.90T>C) located within the first non-coding exon of bovine MLN affects a NKX2-5 transcription factor binding site and showed significant associations (OR(allele) = 0.64; -log(10)P(allele) = 6.8, -log(10)P(genotype) = 7.0) with LDA. An expression study gave evidence of a significantly decreased MLN expression in cows carrying the mutant allele (C). In individuals heterozygous or homozygous for the mutation, MLN expression was decreased by 89% relative to the wildtype. FN298674:g.90T>C may therefore play a role in bovine LDA via the motility of the abomasum. This MLN SNP appears useful to reduce the incidence of LDA in German Holstein cattle and provides a first step towards a deeper understanding of the genetics of LDA.

Approaching the bulk limit with finite cluster calculations using local increments: the case of LiH.

Finite-cluster calculations employing high-level wavefunction-based ab initio methods and extended atomic-orbital basis sets are used to determine local energy increments for bulk LiH. It is shown that these increments can be converged with respect to cluster size and point-charge embedding so as to yield bulk cohesive energies with an accuracy of better than 1 mE(h), both at the Hartree-Fock and at correlated levels. Instrumental for the efficiency of the scheme is the introduction of non-orthogonal orbitals, at an intermediate stage.

Vaccine-induced antibodies linked to bovine neonatal pancytopenia (BNP) recognize cattle major histocompatibility complex class I (MHC I).

A mysterious disease affecting calves, named bovine neonatal pancytopenia (BNP), emerged in 2007 in several European countries. Epidemiological studies revealed a connection between BNP and vaccination with an inactivated vaccine against bovine virus diarrhea (BVD). Alloantibodies reacting with blood leukocytes of calves were detected in serum and colostrum of dams, which have given birth to calves affected by BNP. To understand the linkage between vaccination and the development of alloantibodies, we determined the antigens reacting with these alloantibodies. Immunoprecipitation of surface proteins from bovine leukocytes and kidney cells using sera from dams with a confirmed case of BNP in their gestation history reacted with two dominant protein species of 44 and 12 kDa. These proteins were not detected by sera from dams, free of BVDV and not vaccinated against BVD, and from sera of animals vaccinated with a different inactivated BVD vaccine. The 44 kDa protein was identified by mass spectrometry analysis as MHC I, the other as ß-2-microglobulin. The presence of major histocompatibility complex class I (MHC I) in the vaccine was confirmed by Western blot using a MHC I specific monoclonal antibody. A model of BNP pathogenesis is proposed.

Detection of colostrum-derived alloantibodies in calves with bovine neonatal pancytopenia.

Bovine neonatal pancytopenia (BNP) is an emerging calf disease of unknown cause characterized by a pronounced susceptibility to bleeding as a result of a pancytopenia and bone marrow depletion. In this study we investigated whether this phenomenon is related to colostrum-derived alloantibodies directed against neonatal leukocytes. In a first experiment and using a flow cytometric approach sera from 6 BNP-dams (had given birth to BNP-calves; vaccinated against bovine viral diarrhea virus [BVDV]) and 6 control-dams (no herd history of BNP; no BVDV vaccination) were analyzed for the presences of alloantibodies (IgG) able to bind to the surface of leukocytes isolated from 7 calves from a herd with no history of BNP (no BVDV vaccination). In a second experiment, 4 neonates from 3 BNP-dams were fed colostrum from their corresponding mothers and sampled on a regular basis from birth up to day 21 of life under clinically controlled conditions. Sample analysis of the 4 neonates included hematology (white blood cell count and platelets), bone marrow cytology and histopathology as well as the flow cytometric detection of the percentage of IgG+-lymphocytes/monocytes in the peripheral blood. Experiment #1 showed that all BNP-dam sera harbored significantly higher alloantibody titers than the control dam sera (p<0.001). In the peripheral blood of the two neonates (Experiment #2), the percentage of IgG+-cells increased dramatically within 12h post colostrum intake (p.c.i.), remaining at over 95% for up to 3 days. Both calves developed BNP-associated clinical symptoms, one died. Both twin calves showed no clinical symptoms accompanied by a minor increase of IgG+ cells for up to 12h. Thus, the level of IgG+-cells and the duration of the detection thereof correlated with the severity of BNP developed by these animals. The results show that BNP-dams harbor alloantibodies against surface antigens of neonatal leukocytes in their sera that are readily transferred to the offspring via colostrum. These alloantibodies probably play a crucial role in the pathogenesis of BNP.

Differential proteome analysis of Mycobacterium avium subsp. paratuberculosis grown in vitro and isolated from cases of clinical Johne's disease.

Bovine Johne's disease (paratuberculosis), caused by Mycobacterium avium subspecies paratuberculosis, poses a significant economic problem to the beef and dairy industry worldwide. Despite its relevance, however, pathogenesis of Johne's disease is still only partially resolved. Since mycobacterial membrane proteins expressed during infection are likely to play an important role in pathogenesis, membrane-enriched fractions, namely mucosa-derived membranes (MDM) and culture-derived membranes (CDM), of M. avium subsp. paratuberculosis from three cows with clinical paratuberculosis were investigated. An initial analysis by 2D difference gel electrophoresis (2D DIGE) and MALDI-TOF-MS analysis revealed four differentially expressed proteins with only one predicted membrane protein. Due to this limited outcome, membrane preparations were subjected to a tube-gel trypsin digestion and investigated by using nanoflow-liquid-chromatography-coupled tandem MS. Based on this approach a total of 212 proteins were detected in MDM including 32 proteins of bovine origin; 275 proteins were detected in CDM; 59 % of MDM and CDM proteins were predicted to be membrane-associated. A total of 130 of the proteins were detected in both MDM and CDM and 48 predicted membrane proteins were detected in MDM from at least two cows. Four of these proteins were not detected in CDM, implying differential expression in the host. All membrane-associated proteins, especially the four identified as being differentially expressed, might be relevant targets for further analyses into the pathogenesis of bovine paratuberculosis.

Addressing chemical diversity by employing the energy landscape concept.

Exploring the structural diversity of a chemical system rests on three pillars. First, there is the global exploration of its energy landscape that allows one to predict which crystalline modifications can exist in a chemical system at a given temperature and pressure. Next, there is the development of new synthesis methods in solid-state chemistry, which require only very low activation energies such that even metastable modifications corresponding, for example, to minima on the landscape surrounded by low barriers can be realized. Finally, there is the theoretical design of optimal synthesis routes, again based on the study of the system's energy landscape. In this paper the energy landscape approach to the prediction of stable and metastable compounds as a function of temperature and pressure is presented, with a particular focus on possible phase transitions. Furthermore, several examples are presented, where such predicted compounds were subsequently successfully synthesized, often employing a newly developed synthesis method, low-temperature atom-beam deposition.

Congenital infiltrative lipomas in a calf.

The current report describes a case of congenital subcutaneous and intramuscular tumors of the neck and tail base in a 4-week-old female Angus-Charolais crossbred calf. Results of clinical and ultrasound examination are summarized. Biopsy and necropsy findings indicated an infiltrative lipoma. Congenital lipomas are uncommon tumors in bovids. Clinical and morphologic differentials, as well as classification and the possible pathogenesis of congenital neoplasms, are discussed.