Precalving behavior in dairy cattle with different calving times.

Predicting the calving time in dairy cattle can help in avoiding calving accidents and reducing burdens on animal caretakers. In this study, we analyzed the behavior of pregnant dairy cattle for 7 days prior to calving, to assess the feasibility of predicting the calving time. Eleven Holstein cows were divided into two groups based on their calving times, that is, in the morning (the Morning Parturition Group) or the evening (the Evening Parturition Group). Their behavior was recorded on video. An analysis was conducted of the daily occurrences of each type of behavior and the number of switches of behavior during the day and at night. A statistical analysis was conducted, using a two-way factorial analysis. The behavioral sequence was analyzed using an adjacency matrix. Hierarchical structure charts were created, using Interpretive Structural Modeling. The results suggest that feeding and exploratory behaviors are associated with the calving time period and thus can be useful when predicting that period. The hierarchical structure charts suggest that the Morning Parturition Group had no definite behavioral sequence pattern, unlike the Evening Parturition Group. The detection of an unstable behavioral sequence pattern might predict the calving time period.

The effect of exposure on cattle thyroid after the Fukushima Daiichi nuclear power plant accident.

Nuclear plant accidents can be a risk for thyroid cancer due to iodine radioisotopes. Near the Fukushima Daiichi nuclear power plant, cattle were exposed to radiation after the accident occurred in May 2011. Here we estimated the total radiation exposure to cattle thyroid and its effects on thyroid function. Until October 2016, the estimated external exposure dose in Farm A was 1416 mGy, while internal exposure dose of 131I, 134Cs, and 137Cs were 85, 8.8, and 9.7 mGy in Farm A and 34, 0.2, and 0.3 mGy in Farm B, respectively. The exposed cattle had thyroid with relatively lower weight and lower level of stable iodine, which did not exhibit any pathological findings. Compared with the control, the plasma level of thyroid-stimulating hormone (TSH) was higher in Farm A cattle born before the accident, while the plasma thyroxine (T4) was higher in Farm A cattle born after the accident, suggesting that exposed cattle showed slight hyperactivation of the thyroid gland. In addition, Farm A cattle have higher level of cortisol, one of the anterior pituitary gland-derived hormones. However, we did not observe a causal relationship between the radiation exposure and cattle thyroid.

A Molecule Having 13 Unpaired Electrons: Magnetic Property of a Gadolinium(III) Complex Coordinated with Six Nitronyl Nitroxide Radicals.

A gadolinium(III) complex coordinated with six nitronyl nitroxide radicals showed intriguing temperature-dependent changes in magnetic susceptibilities. The gadolinium(III) ion in the complex is pseudo-eight-coordinated by three singlet-ground-state diradical anion species based on nitronyl nitroxide radicals. The magnetic susceptibility (χpT) of the gadolinium(III) complex at 298 K, whose value corresponded to that of a system with 13 unpaired electrons (seven-spin system), decreased upon a lowering of the temperature to 11 K but increased upon a further lowering of the temperature. Finally, the χpT value at 2 K was found to be higher than that at room temperature. The temperature-dependent magnetic behavior could be explained by a structural change in the diradical anion ligand due to its flexibility.

Smallest Organic Tetracation in the Gas Phase: Stability of Multiply Charged Diiodoacetylene Produced in Intense Femtosecond Laser Fields.

Coulomb explosion imaging, which is the reconstruction of a molecular structure by measuring the three-dimensional momenta of atomic ions formed by a Coulomb explosion of multiply charged molecular cations (MMCs), has been utilized widely. In contrast, intact MMCs, whose properties and reactions are interesting from both fundamental and applied scientific perspectives, themselves have been little explored to date. This study demonstrates that the four-atom molecule diiodoacetylene (DIA) can survive as a long-lived species in the gas phase after the removal of four electrons in intense femtosecond laser fields. The electron configurations of the equilibrium structures of the electronic ground states calculated by the complete active space self-consistent field (CASSCF) method reveal the stability of multiply charged DIA. The dissociation energies are estimated to be 3.01, 3.59, 2.57, 1.82, and 1.61 eV for neutral, cation radical, dication, trication radical, and tetracation, respectively. A fairly deep potential well suggests that a DIA tetracation is metastable toward dissociation, whereas the repulsive potential of a pentacation radical confirms its absence in the mass spectrum. With their sufficiently long lifetimes, minimum number of atoms, and simple dissociation paths, DIA MMCs are promising candidates for further experimental and theoretical investigations of multiply charged ion chemistry.

Neu1 deficiency induces abnormal emotional behavior in zebrafish.

NEU1 sialidase hydrolyzes sialic acids from glycoconjugates in lysosomes. Deficiency of NEU1 causes sialidosis with symptoms including facial dysmorphism, bone dysplasia, and neurodegeneration. However, the effects of NEU1 deficiency on emotional activity have not been explored. Here, we conducted the behavioral analysis using Neu1-knockout zebrafish (Neu1-KO). Neu1-KO zebrafish showed normal swimming similar to wild-type zebrafish (WT), whereas shoaling was decreased and accompanied by greater inter-fish distance than WT zebrafish. The aggression test showed a reduced aggressive behavior in Neu1-KO zebrafish than in WT zebrafish. In the mirror and 3-chambers test, Neu1-KO zebrafish showed more interest toward the opponent in the mirror and multiple unfamiliar zebrafish, respectively, than WT zebrafish. Furthermore, Neu1-KO zebrafish also showed increased interaction with different fish species, whereas WT zebrafish avoided them. In the black-white preference test, Neu1-KO zebrafish showed an abnormal preference for the white region, whereas WT zebrafish preferred the black region. Neu1-KO zebrafish were characterized by a downregulation of the anxiety-related genes of the hypothalamic-pituitary-adrenal axis and upregulation of lamp1a, an activator of lysosomal exocytosis, with their brains accumulating several sphingoglycolipids. This study revealed that Neu1 deficiency caused abnormal emotional behavior in zebrafish, possibly due to neuronal dysfunction induced by lysosomal exocytosis.

Aurophilic Interactions in Multi-Radical Species: Electronic-Spin and Redox Properties of Bis- and Tris-[(Nitronyl Nitroxide)-Gold(I)] Complexes with Phosphine-Ligand Scaffolds.

Multinuclear AuI complexes with two or three nitronyl nitroxide-2-ide radical anion and phosphine-ligand scaffolds, (NN-Au)2 -1 o, (NN-Au)2 -1 m, and (NN-Au)2 -1 p, have been synthesized to investigate the influence of AuI -AuI (aurophilic) interactions on the properties of multispin molecular systems. The desired complexes were successfully prepared in moderate yields in a one-pot synthesis from the corresponding phosphine ligand, AuI source, parent NN, and sodium hydroxide. Among the prepared complexes, (NN-Au)2 -1 o, in which an aurophilic interaction was clearly observed by crystal structure analysis, showed characteristic spin-spin interactions, electrochemical properties, and solvatochromic behavior. The results from theoretical calculations also suggested that the differences in properties between complex (NN-Au)2 -1 o and the other complexes are due to intramolecular aurophilic interactions.

Syntheses and Properties of (Nitronyl nitroxide)-substituted Tri-phenylamine ortho-Bridged by Two Oxygen and Sulfur Atoms.

A nitronyl nitroxide unit (NN) was linked with a triphenylamine-based condensed polycyclic skeleton DOTT to form a radical substituted donor NN-DOTT. X-ray crystal structure analysis demonstrated a flat bowl shape of the DOTT unit. EPR spectra showed the localization of electron spin on the NN unit. Chemical oxidation of the DOTT unit produced radical-substituted radical cation salts NN-DOTT+ ⋅ SbF6 - and NN-DOTT+ ⋅ FeBr4 - that are stable under ambient conditions. The magnetic behavior of NN-DOTT+ ⋅ SbF6 - is characterized by the strong intramolecular ferromagnetic interaction between NN and DOTT+ . The X-ray structural analysis of NN-DOTT+ ⋅ FeBr4 - shows planar structure of DOTT and 1D mixed-stack column of NN-DOTT+ and FeBr4 - . Magnetic measurements established that NN-DOTT+ ⋅ FeBr4 - undergoes magnetic phase transition into a weak ferromagnet at 7 K.

Assessments of DNA Damage and Radiation Exposure Dose in Cattle Living in the Contaminated Area Caused by the Fukushima Nuclear Accident.

Since the Fukushima nuclear accident in 2011, various abnormalities have been reported in animals living in the contaminated area. In the present study, we examined DNA damage in cattle living in the "difficult-to-return zone" by 8-hydroxy-2'-deoxyguanosine, comet, and micronucleus assays using their peripheral blood. The radiation exposure dose rate at the sampling time was approximately 0.25 or 0.38 mGy/day and the cumulative dose was estimated at approximately 1000 mGy. Significant increase in DNA damage was not detected by any of the three methods. As DNA damage is a stochastic effect of radiation, it might be occurring in animals living in the contaminated area. However, the present results suggest that radiation-induced DNA damage in the cattle did not increase to the level detectable by the assays we used due to the low dose rate in this area.

Condensed Phenoxazine Dimer and Its Radical Cation.

A condensed phenoxazine dimer was synthesized and characterized. X-ray crystallographic analysis of the dimer shows a double-butterfly structure, in which the nitrogen atoms are located above and below the molecular plane. A radical cation salt of the dimer was obtained using tris(4-bromophenyl)aminium hexafluoroantimonate as the oxidant. The salt is air-stable in solid and solution states. The cation structure was evaluated by X-ray crystallographic analysis, showing that the phenoxazine units were converted to a planar structure upon oxidation.

Establishment and characterization of Neu1-knockout zebrafish and its abnormal clinical phenotypes.

Mammalian sialidase Neu1 is involved in various physiological functions, including cell adhesion, differentiation, cancer metastasis, and diabetes through lysosomal catabolism and desialylation of glycoproteins at the plasma membrane. Various animal models have been established to further explore the functions of vertebrate Neu1. The present study focused on zebrafish (Danio rerio) belonging to Cypriniformes as an experimental animal model with neu1 gene deficiency. The results revealed that the zebrafish Neu1 desialyzed both α2-3 and α2-6 sialic acid linkages from oligosaccharides and glycoproteins at pH 4.5, and it is highly conserved with other fish species and mammalian Neu1. Furthermore, Neu1-knockout zebrafish (Neu1-KO) was established through CRISPR/Cas9 genome editing. Neu1-KO fish exhibited slight abnormal embryogenesis with the accumulation of pleural effusion; however, no embryonic lethality was observed. Although Neu1-KO fish were able to be maintained as homozygous, they showed smaller body length and weight than the wild-type (WT) fish, and muscle atrophy and curvature of the vertebra were observed in adult Neu1-KO fish (8 months). The expression patterns of myod and myog transcription factors regulating muscle differentiation varied between Neu1-KO and WT fish embryo. Expression of lysosomal-related genes, including ctsa, lamp1a, and tfeb were up-regulated in adult Neu1-KO muscle as compared with WT. Furthermore, the expression pattern of genes involved in bone remodeling (runx2a, runx2b, and mmp9) was decreased in Neu1-KO fish. These phenotypes were quite similar to those of Neu1-KO mice and human sialidosis patients, indicating the effectiveness of the established Neu1-KO zebrafish for the study of vertebrate Neu1 sialidase.

16S rRNA-based amplicon analysis of changes in the bacterial population in the lesions of papillomatous digital dermatitis in dairy cattle after topical treatment with allyl isothiocyanate.

Papillomatous digital dermatitis (PDD) is a foot disease causing lameness in dairy cattle. It is regarded as a polymicrobial infection, although its etiology is not fully understood. PDD is treated by the topical or systemic administration of antibiotics such as lincomycin (LCM); however, the milk of the cows cannot be marketed during the treatment and withdrawal period due to the residual antibiotics in milk. Allyl isothiocyanate (AITC), an extract of Wasabia japonica (known as wasabi or Japanese horseradish) widely employed as a food additive, can be used as an alternative antimicrobial agent that overcomes this problem. We previously showed that AITC is as effective as LCM in PDD treatment. Here, using the samples obtained in the previous clinical study, we analyzed changes in the bacterial population in the PDD-associated microbiota after AITC treatment and compared those with that following LCM treatment by 16S ribosomal RNA (rRNA)-based amplicon analysis. Both treatments induced major changes in the bacterial population, and Treponema species, which have been regarded as the major causative agents of PDD, were efficiently eliminated by both agents. However, the AITC-treated samples exhibited higher diversity compared with pretreatment samples, but this trend was not observed for LCM treatment, probably reflecting different antibacterial activities of the two agents. Importantly, this analysis detected population changes before morphological changes in PDD lesions (clinical signs of healing) became evident, indicating that 16S rRNA-based amplicon analysis represents an efficient strategy for analyzing and monitoring the treatment efficiency of PDD as well as other polymicrobial diseases.

Inhibition of Ligand Binding Ability of Three Porphyrins by an Organic Effector.

A stimulus-responsive receptor 1 was designed and prepared to control the ligand-binding ability of three active sites, two zinc tetraphenylporphyrin units (P1) and one zinc diethynyldiphenylporphyrin unit (P2), with one effector molecule 2. Bulky hexarylbenzene units were incorporated as shielding panels in the middle of the flexible side arms of 1. Spectroscopic titrations indicated that a stable supramolecular complex 1⋅2 (K1⋅2 =6.7×106 m-1 ) was produced by the cooperative formation of multiple hydrogen and coordination bonds. As a result, the binding of a ligand to P1 was inhibited by 2 in a competitive manner. Additionally, the formation of 1⋅2 brought about conformational restriction of the side arms to cover both faces of P2 with the shielding panels. The binding constant of 4-phenylpyridine with P2 in 1⋅2 decreased to 8.9 % of that in 1. Namely, the ligand-binding ability of P2 was inhibited according to an allosteric mechanism.

Redox-Induced Modulation of Exchange Interaction in a High-Spin Ground-State Diradical/Triradical System.

A triplet ground-state diradical molecule, bis(nitronyl nitroxide)-substituted diphenyldihydrophenazine (1.. ), that can be converted into a one-electron oxidized species, 1…+ , in the quartet ground state has been developed. Surprisingly, these species, 1.. and 1…+ , can be used under ambient conditions because they are reasonably stable under aerobic conditions, even in solution. The temperature-dependent magnetic susceptibilities reveal that 1.. and 1…+ are in the triplet state, with a weak exchange interaction (J1 /kB = +3.1 K) and quartet ground state with a strong exchange interaction (J2 /kB = +160 K), respectively. The interconversion between the neutral and one-electron oxidized species can be realized through electrochemical reactions. Significantly different absorption bands in the near-IR region newly appeared in the electronic spectra acquired during electrochemical oxidation/reduction.

Spin-dependent electron transfer dynamics in a platinum-complex-donor-acceptor triad studied by transient-absorption detected magnetic field effect.

For realization of efficient organic light-energy conversion systems, controlling the lifetime of photogenerated charge separated states in donor (D)-acceptor (A) molecules is of much importance; the spin dynamics is one of the important controlling factors. We previously reported that the covalently-linked 1,3-bis(2-pyridylimino)-isoindolate platinum (BPIPt)-dimethoxytriphenylamine (D)-naphthaldiimide (A) triad molecule (BPIPt-DA) exhibits a triplet-born long-lived charge separated state (BPIPt-D•+A•-), the lifetime of which is significantly increased from 4 µs to 10 µs by an applied magnetic field of 270 mT in room temperature tetrahydrofuran (THF). The purpose of the present study is to clarify detailed dynamics of spin-dependent generation and the decay of BPIPt-D+A-. For this purpose, we measured transient optical absorption (TA) and the TA-detected magnetic field effect (MFE) as functions of temperature and dispersion media. In THF at 183 K, MFE-detected transient spectra of the intermediate BPIPt•--D•+A state are observed. We have successfully quantified the recombination loss at this state by a kinetic simulation of MFE without using any reference molecules. The lifetime of the final BPIPt-D•+A•- state in a cellulose acetate polymer matrix at room temperature is significantly prolonged to 20 µs at 0 mT and 96 µs at 250 mT compared to those in THF. From the comparison of temperature dependences of the two media, effects of molecular motions on the electronic coupling and the spin relaxation are discussed.

Pathological characteristics of thyroid glands from Japanese Black Cattle living in the restricted area of the Fukushima Daiichi Nuclear Power Plant accident.

To study the effect of ionizing radiation on thyroid glands, 66 Japanese Black cattle residing in the restricted area of the Fukushima Daiichi Nuclear Power Plant in 2013-2017 were pathologically examined. There were no clinical symptoms of thyroid disease in these cattle. Three cases of goiter and seven of atrophy were found in two among the four farms examined. Cases of goiter exhibited normal morphological structure without mass or nodule formation in thyroid glands. Cellular atypia or capsular invasion of the follicular epithelium was absent. The estimated integrated dose of external radiation in goiter cases ranged from maximum 797 mSv to minimum 24 mSv. All lobules in the seven atrophic thyroid glands were affected, but pathological findings, such as inflammatory cell infiltration or stromal fibrosis, were not observed. The estimated integrated dose of external radiation in atrophic thyroids ranged from maximum 589 mSv to minimum 8 mSv. Immunohistochemical analysis of anti-nitroguanosine and the TUNEL method in goiter and atrophic thyroid glands did not reveal any positive findings. The present study indicates that there was no significant relationship between a radiation effect and pathological findings in any thyroid glands.

Comparison of urine and blood as a convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium.

Blood and urine were compared to clarify which is a better sample for estimating the concentration of radioactive cesium in muscles of live cattle. The most probable concentration of 137 Cs in muscles was 21.0 times that in blood, and the error of this estimation was 28%. The concentration of 137 Cs in blood was estimated accurately using urine samples. The estimation error did not increase so much (33%), even when the concentration of 137 Cs in muscles was estimated using urine samples. On the other hand, the maximum volume of blood that can be collected with one syringe is 50 ml, whereas it is easy to collect 1,000 ml of urine. It took at least 360 min to confirm that a cow meets the legal standard by inspecting 50 ml of blood. However, with 1,000 ml of urine, a 20 min measurement time was sufficient for this purpose. This difference in the required measurement time is critically important for practical use. In addition, urine can be collected by farmers themselves, whereas the blood collection requires a veterinarian. Therefore, urine is a more convenient and practical sample for estimating the contamination level of live cattle with radioactive cesium.

Triplet Diradical-Cation Salts Consisting of the Phenothiazine Radical Cation and a Nitronyl Nitroxide.

The spin-spin and magnetic properties of two (nitronyl nitroxide)-(di-p-anisylamine-phenothiazine) diradical cation salts, (DAA-PTZ)+ -NN⋅MBr4 - (M=Ga, Fe), have been investigated. These diradical-cation species were prepared by the cross-coupling of iodophenothiazine DAA-PTZ-I with NN-AuPPh3 followed by oxidation with the thianthrenium radical cation (TA+ ⋅MBr4 - ). These salts were found to be highly stable under aerobic conditions. For the GaBr4 salt, large ferromagnetic intramolecular and small antiferromagnetic intermolecular interactions (J1 /kB =+320 K and J2 /kB =-2 K, respectively) were observed. The magnetic property of the Fe3+ salt was analyzed by using a six-spin model assuming identical intramolecular exchange interaction (J3 /kB =+320 K) and the other exchange interactions (J4 /kB =-7 K and J5 /kB =-4 K). A significant color change was observed in the UV/Vis/NIR absorption spectra upon electrochemical oxidation of the doublet DAA-PTZ-NN to the triplet (DAA-PTZ)+ -NN.

Effects of Treatment Time and Thickness of Meat on the Removal of Radioactive Cesium from Beef Slices by Boiling and Water Extraction.

HIGHLIGHTS: Boiling effectively decreased radioactive cesium concentrations in beef. Water extraction was also effective but not as effective as boiling. The decontamination effect increased with treatment time. The decontamination effect was inversely proportion to the square of the thickness of the beef slice.

Dairy cattle behavior classifications based on decision tree learning using 3-axis neck-mounted accelerometers.

Demand has been increasing recently for an automated monitoring system of animal behavior as a tool for the management of livestock animals. This study investigated the association between the behavior of dairy cattle and the acceleration data collected using three-axis neck-mounted accelerometers, as well as the feasibility of improving the precision of behavior classifications through machine learning. In total 38 Holstein dairy cows were used, and kept in four different farms. A logger was mounted to each collar to obtain acceleration data for calculating the activity level and variations. At the same time the behavior of the cattle was observed visually. Characteristic acceleration waves were recorded for eating, rumination, and lying, respectively; and the activity level and variations were significantly different among these behaviors (p < 0.01). Decision tree learning was performed on the data set from Farm A and validated its precision; which proved to be 99.2% in cross-validation, and 100% in test data sets from Farms B to D. This study showed that highly precise classifications for eating, rumination, and lying is possible by using decision tree learning to calculate the activity level and variations of cattle based on the data obtained by three-axis accelerometers mounted to a collar.

Synthesis and Properties of Nitrogen-Containing Pyrenes.

We developed a short-step synthesis of 2,4,10-triazapyrenes involving two sequential C-H substitutions: Pd-catalyzed cross-coupling reactions via C-H arylation followed by intramolecular Cu-catalyzed C-H functionalization. This method was successfully applied to the preparation of 4,10-diaza-, 1,4,10-triaza-, and 1,3,4,10-tetraazapyrenes. Crystal structure analysis of 5,9-di(4-methylphenyl)-2,4,10-triazapyrene showed that planar triazapyrene cores have π-stack packing. Incorporating nitrogen atoms into the pyrene framework bathochromically shifted the lowest energy onsets of the absorption bands and increased the first reduction potentials. The nitrogen-containing pyrenes showed fluorescence with a weaker intensity (Φf = 0.041-0.12) than that of the parent pyrene. The number and position of nitrogen atoms influenced the extent of these effects.