Divergent ornamentation within a single population of the barn swallow.

Differential migration strategies favour different sets of characteristics, including sexually selected ornamentation. Such phenotypic variation is particularly evident in a population with partial migration, where migrants and non-migrants co-exist. Partial migration provides insights into the link between migration, local environment and ornamentation, although empirical studies remain scarce. Here, we studied the plumage traits of barn swallows (Hirundo rustica) in southern Japan, where both winterings and migrants breed sympatrically. We further examined this relationship with multiple isotopes (δ2H, δ13C, δ15N and δ34S), which provides insight into their moulting habitat. Among males, winterings and migrants differed in their morphological traits: wintering males had shorter wings, which suggests the high demand for flight apparatus in migratory birds. Moreover, wintering males had larger white tail spots and less colourful throat patches than migratory males, indicating ornament divergence between them. Wintering males had a significantly smaller isotopic space when examining the combinations of δ34S with the other isotopes compared to migratory males, which indicates a differential geographic range between them, perhaps because of the limited variation in the distance to the sea in wintering males. As in males, wintering females had a significantly smaller isotopic space than migrant females, but there were few morphological differences between migratory and wintering females. Instead, some morphological traits were related to isotope values in females. These results indicate sex-specific linkage between migration, local environment, and ornamentation.

Identification of a Weak Interaction between the Spin-Only 4f-Electronic System of Trivalent Gd Ion and a Photoexcited Cyclic π Electronic System.

An electronic interaction between the spin-only 4f-electronic system and the photoexcited π cyclic system has been identified in the anionic bis(phthalocyaninato)gadolinium(III) complex [Pc2Gd]- by using variable-temperature variable-magnetic-field magnetic circular dichroism (VTVH MCD). Two positive MCD A-term patterns, corresponding to the QH and QL absorption bands, were observed to increase in intensity as the temperature decreased, indicating a ferromagnetic-type interaction between spin angular momentum S of the 4f system and orbital angular momentum L of the photoexcited π system. A theoretical model incorporating the S-L interaction constant CSL was fitted to the VTVH MCD spectra, confirming the presence of a weak interaction, whose intensity was determined as CSL = 0.27 cm-1 for QL and CSL = 0.02 cm-1 for QH. Ab initio calculations using the RASSCF and RASSI methods revealed that the inclusion of the excited state calculations with the spin multiplicity (2S + 1) of 6 was crucial for assigning ΔSL and CSL values. This underscores the essential role of the 6PJ term in accurately modeling the S-L interaction in [Pc2Gd]-.

Electronic structure analysis of phthalocyanine complexes using magnetic circular dichroism and magnetic circularly polarized luminescence spectroscopy.

In the study of phthalocyanine complexes using magnetic circular dichroism (MCD) spectroscopy, the electronic structure of excited states is generally discussed based on the rigid-shift approximation, in which the band profiles for left-handed circularly polarized (lcp) and right-handed circularly polarized (rcp) light are assumed to be the same. This assumption may not necessarily be valid for cases where there are multiple initial states having different geometries. Magnetic circularly polarized luminescence (MCPL) from phthalocyanine complexes can be regarded as an example of such cases, since the two degenerate emission states are split in a magnetic field and can undergo a structural deformation. Here, we investigated an alternative approach, where the lcp and rcp components are independently determined. This method, which we refer to as the direct-separation approach, allows direct determination of the distribution of the two emission states as well as the orbital angular momentum L z $$ \left|{L}_z\right| $$ . Using this approach, L z $$ \left|{L}_z\right| $$ and the distribution were determined from MCD and MCPL spectra of a series of phthalocyanine complexes. Comparison of the two methods shows that the rigid-shift and the direct-separation approaches give practically equivalent results for the systems under study, but the latter is advantageous for systems where the former is not applicable.

Construction of high-throughput magnetic circular dichroism measurement system and its application to research on magnetic and optical properties of phthalocyanine complexes.

Magnetic circular dichroism (MCD) spectroscopy is a powerful method for evaluating the electronic structure and magnetic and optical properties of molecules. In particular, MCD measurements have been performed on phthalocyanines and porphyrins with various central metal ions, axial ligands, and substituents to elucidate their properties. It is essential to develop a robust high-throughput technique to perform these measurements comprehensively and efficiently. However, MCD spectroscopy requires very high optical quality for each component of the instrument, and even slight cell distortions can impair the baseline flatness. Consequently, when versatility and data quality are important, an optical system designed for a microplate reader is not suitable for the MCD spectrometer. Therefore, in this study, we develop a new magnetic flow-through cell and combine it with an existing CD spectrometer and autosampler to construct a high-throughput system. The effectiveness and performance of this new system are then evaluated. In addition, based on the MCD and absorption spectra of various phthalocyanine complexes, the effects of substituents and solvents on their magnetic and optical properties and the causes of these effects are discussed. The results demonstrate that this system is effective for the evaluation of the physicochemical properties of various phthalocyanine complexes.

Application of a porous graphitic carbon column to carbon and nitrogen isotope analysis of underivatized individual amino acids using high-performance liquid chromatography coupled with elemental analyzer/isotope ratio mass spectrometry.

RATIONALE: Isolation of underivatized amino acids (AAs) using high-performance liquid chromatography (HPLC) is becoming a popular method for carbon (δ13 C) and nitrogen isotope (δ15 N) analyses of AAs because of the high analytical precision and for performing dual-isotope analysis. However, some AAs in natural samples, especially small, hydrophilic AAs, are not suitably separated using reversed-phase columns (e.g., C18) and ion-exchange columns (e.g., Primesep A). METHODS: We developed a new method for HPLC using a porous graphitic carbon column for the separation of nine hydrophilic AAs. After purification, δ13 C and δ15 N values of AAs were determined using elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). We demonstrated the application of this method by determining δ13 C and δ15 N values of individual hydrophilic AAs in a biological sample, the muscle of blue mackerel (Scomber australasicus). RESULTS: Chromatographically, the baseline separation of hydrophilic AAs was achieved in both the standard mixture and the biological sample. We confirmed that δ13 C and δ15 N values of AA standards remained unchanged during the whole experimental procedure. The δ13 C values of AAs in mackerel muscle are also in good agreement with the values obtained using another verified method for δ13 C analysis. CONCLUSIONS: The good separation performance of hydrophilic AAs and the reliability of δ13 C and δ15 N analyses of individual AAs using the porous graphite column offer a significant advantage over conventional settings. We suggest that, in the future, the HPLC × EA/IRMS method can be used for reliable δ13 C and δ15 N analyses of AAs in natural samples.

Janus Pyrrolopyrrole Aza-dipyrrin: Hydrogen-Bonded Assemblies and Slow Magnetic Relaxation of the Cobalt(II) Complex in the Solid State.

A novel pyrrolopyrrole azadipyrrin (Janus-PPAD) with Janus duality was synthesized by a Schiff base-forming reaction of diketopyrrolopyrrole. The orthogonal interactions of the hydrogen-bonding ketopyrrole and metal-coordinating azadipyrrin moieties in Janus-PPAD enabled the metal ions to be arranged at regular intervals: zinc(II) and cobalt(II) coordination provided metal-coordinated Janus-PPAD dimers, which can subsequently form hydrogen-bonded one-dimensional arrays both in solution and in the solid state. The supramolecular assembly of the zinc(II) complex in solution was investigated by 1 H NMR spectroscopy based on the isodesmic model, in which a binding constant for the elongation of assemblies is constant. Owing to the tetrahedral coordination, in the solid state, the cobalt(II) complex exhibited a slow magnetic relaxation due to the negative D value of -27.1 cm-1 with an effective relaxation energy barrier Ueff of 38.0 cm-1 . The effect of magnetic dilution on the relaxation behavior is discussed. The relaxation mechanism at low temperature was analyzed by considering spin lattice interactions and quantum tunneling effects. The easy-axis magnetic anisotropy was confirmed, and the relevant wave functions were obtained by ab initio CASSCF calculations.

Antiparallel Coupling between a 4f System and a Photoexcited Cyclic π System in a Dysprosium(III) Monoporphyrinato Complex.

Magnetic coupling resulted by an orbital angular momentum L of a photoexcited cyclic π-conjugated system and total angular momentum J of a localized 4f electronic system in [Dy(TPP)(cyclen)]Cl (TPP = 5,10,15,20-tetraphenylporphyrinato; cyclen = 1,4,7,10-tetraazacyclododecane) has been identified using magnetic circular dichroism (MCD). The MCD A-term patterns of the compound show a negative increment in the intensity by decreasing temperature and nonproportionality to the magnetic field. This phenomenon is experimentally observed for the first time in porphyrin-based lanthanide complexes and indicates the presence of the J-L interaction, with the preferred orientation being antiparallel. A theoretical model in which J and L are assumed to couple antiferromagnetically yielded a quantitative agreement to the experimental data of varied-temperature and varied-magnetic-field MCD spectral measurements. The ab initio calculations were performed, revealing the relationship between two orbital angular momenta, i.e., L(π) and L(f), which result from the delocalized π system and the localized 4f system, respectively.

Intramolecular Magnetic Interaction in a Photogenerated Dual Angular Momentum System in a Terbium-Phthalocyaninato 1:1 Complex.

Intramolecular magnetic interaction between a localized open-shell 4f-electronic system and a photoexcited macrocyclic π-conjugate system in terbium-phthalocyaninnato (Tb-Pc) 1:1 complex was investigated using variable-temperature variable-field magnetic circular dichroism (VTVH MCD) spectroscopy. The 1:1 complex [Tb(Pc)(cyclen)]Cl (Pc2- = phthalocyaninato dianion, cyclen = 1,4,7,10-tetraazacyclododecane) with the capping ligand providing an exact fourfold symmetry showed a significant temperature dependence and a nonlinear field dependence in the MCD intensity of the Pc-centered highest occupied molecular orbital (HOMO)-lowest unoccupied molecular orbital (LUMO) π-π* transition, while a diamagnetic congener [Y(Pc)(cyclen)]Cl showed a temperature-independent MCD with a linear-field dependence. This indicates that the (4f)8 system of the Tb ion with a total angular momentum J and the photoexcited π-system of the Pc macrocycle with an orbital angular momentum L are magnetically coupled. By numerical simulation using a model where ground doublet state |Jz⟩ = |±6⟩ and excited quartet state |Jz, Lz⟩ = |±6, ±|Lz|⟩ are included, the J-L interaction magnitude ΔJL and the Pc-centered orbital angular momentum |Lz|ℏ were determined to be 1.1 cm-1 and 2.0 ℏ, respectively. From ab initio restricted active space self-consistent field (RASSCF)-restricted active space state interaction (RASSI) calculations on the π-π* excited states of the Tb complex, the magnitude of the J-L interaction was estimated. The comparison between the calculations on the Y and Tb complexes revealed that the ferromagnetic-type coupling occurs between the orbital component in the J of Tb and the L on Pc, supporting the model that we employed for the analysis of the experimental data.

Tracking long-distance migration of marine fishes using compound-specific stable isotope analysis of amino acids.

The long-distance migrations by marine fishes are difficult to track by field observation. Here, we propose a new method to track such migrations using stable nitrogen isotopic composition at the base of the food web (δ15 NBase ), which can be estimated by using compound-specific isotope analysis. δ15 NBase exclusively reflects the δ15 N of nitrate in the ocean at a regional scale and is not affected by the trophic position of sampled organisms. In other words, δ15 NBase allows for direct comparison of isotope ratios between proxy organisms of the isoscape and the target migratory animal. We initially constructed a δ15 NBase isoscape in the northern North Pacific by bulk and compound-specific isotope analyses of copepods (n = 360 and 24, respectively), and then we determined retrospective δ15 NBase values of spawning chum salmon (Oncorhynchus keta) from their vertebral centra (10 sections from each of two salmon). We then estimated the migration routes of chum salmon during their skeletal growth by using a state-space model. Our isotope tracking method successfully reproduced a known chum salmon migration route between the Okhotsk and Bering seas, and our findings suggest the presence of a new migration route to the Bering Sea Shelf during a later growth stage.

A method for stable carbon isotope measurement of underivatized individual amino acids by multi-dimensional high-performance liquid chromatography and elemental analyzer/isotope ratio mass spectrometry.

RATIONALE: To achieve better precision and accuracy for δ13 C analysis of individual amino acids (AAs), we have developed a new analytical method based on multi-dimensional high-performance liquid chromatography (HPLC) and elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). Unlike conventional methods using gas chromatography, this approach omits pre-column chemical derivatization, thus reducing systematic errors associated with the isotopic measurement. METHODS: The separation and isolation of individual AAs in a standard mixture containing 15 AAs and a biological sample, spear squid (Heterololigo bleekeri) were performed. AAs were isolated using an HPLC system equipped with a reversed-phase column and a mixed-mode column and collected using a fraction collector. After the chromatographic separation and further post-HPLC purification, the δ13 C values of AAs were measured by EA/IRMS. RESULTS: The complete isolation of all 15 AAs in the standard mixture was achieved. The δ13 C values of these AAs before and after the experiment were in good agreement. Also, 15 AAs in the biological sample, H. bleekeri, were successfully measured. The δ13 C values of AAs in H. bleekeri varied by as much as 30‰ with glycine being most enriched in13 C. CONCLUSIONS: The consistency between the δ13 C values of reference and processed AAs demonstrates that the experimental procedure generates accurate δ13 C values unaffected by fractionation effects and contamination. This method is therefore suitable for δ13 C analysis of biological samples with higher precision than conventional approaches. We propose this new method as a tool to measure δ13 C values of AAs in biological, ecological and biogeochemical studies.

Spectroscopic Investigation of Interaction between the 4f Electronic System and the Photoexcited Cyclic π System in Terbium(III) Monoporphyrinato Complex.

Electronic interaction between the total angular momentum (J) of the 4f electronic system and an angular momentum (L) of the photoexcited cyclic π system in a terbium(III) monoporphyrinato complex with a capping ligand cyclen, [Tb(TPP)(cyclen)]Cl (TPP = 5,10,15,20-tetraphenylporphyrinato; cyclen = 1,4,7,10-tetraazacyclododecane), have been investigated by varied-temperature and varied-magnetic-field magnetic circular dichroism (MCD) spectroscopy. Three MCD A-term patterns which correspond to B(0,0), Q(0,0), and Q(1,0) absorption bands were observed for the complex which was incorporated into a thin film of poly(methyl methacrylate), PMMA, as well as that in solution phase. The A-term patterns in the Soret band region and Q-band region were found to show opposite behavior with changes in temperature and magnetic field. This finding clearly indicated not only the presence of the J-L interaction but also the different directions of the J-L interaction; i.e., J and L interact ferromagnetically in the Q(0,0) and Q(1,0) states, while antiferromagnetically in the B(0,0) state. The magnitude of the interaction was determined by simulation-based fitting to experimental A1/D0 ratios, where A1 refers to the A-term intensity and D0 to the oscillator strength of the band. The first ab initio study was also carried out for further insight into the interaction. From the computational study, the relationship between the orbital angular momentum of π electronic system, L(π), and the orbital angular momentum of the 4f system, L(f), was clearly revealed. A computational approach to determine the magnitude and the direction of J-L interaction was established.

Membrane raft-mediated regulation of glucose signaling pathway leading to acrosome reaction in chicken sperm†.

Despite knowledge that glucose metabolism is essential for the regulation of signaling cascades in the sperm that are pre-assembled into specific areas and function at multistage for fertilization, the physiological roles of glucose in avian sperm are poorly understood. Accumulated results of studies conducted in our laboratory and others indicate that sperm possess membrane microdomains, or membrane rafts, which play important roles in several processes, including the induction of acrosome reaction (AR). When characterizing proteomes associated with chicken sperm rafts, we observed marked enrichment of glucose transporter 3 (GLUT3). Here we show that glucose uptake is mediated by membrane rafts and stimulates AR induction by activating AMP-activated protein kinase (AMPK). Using a specific antibody, we observed that GLUT3 is localized to the entire flagellum and acrosome region and highly associated with membrane rafts. The addition of glucose stimulated AR in a dose-dependent manner without affecting sperm motility. AR and glucose uptake assays were performed using both inhibitors and activators, and demonstrated that glucose-dependent AR results from the activity of a glucose transporter located in membrane rafts and associated with AMPK. To better understand the mechanism of AMPK activation by glucose, we evaluated localization and phosphorylation status of AMPKα, showing that glucose uptake stimulates AMPKα phosphorylation, leading to its complete activation. Together, these results lead us to propose a novel mechanism by which glucose uptake stimulates the AMPK signaling pathway via membrane rafts, resulting in maximal acrosomal responsiveness in avian sperm as migrating upward to a fertilization site.

Synthesis of a Neutral Mononuclear Four-Coordinate Co(II) Complex Having Two Halved Phthalocyanine Ligands That Shows Slow Magnetic Relaxations under Zero Static Magnetic Field.

Syntheses of a novel pseudotetrahedral four-coordinate mononuclear Co(II) complex that has two halved phthalocyanine moieties as the ligands, [Co(half-Pc)2] (1), and its magnetic properties as a single molecule magnet (SMM) are reported. A one pot reaction of phthalonitrile and lithium methoxide followed by the coordination to a Co(II) ion gave 1 as an orange solid in a moderate yield. X-ray crystallography on 1 reveals tetragonally distorted coordination geometry around the Co(II) ion. The M- HT-1 plots suggest that 1 has large axial magnetic anisotropy. The ac magnetic susceptibility data of the magnetically diluted 1 (dil.1) clearly show that the complex acts as an SMM even in the absence of the external static magnetic field ( Hdc). The influence of intermolecular and intramolecular interactions for the magnetic relaxation behaviors has been discussed by comparison of the magnetic data of 1 and dil.1. The Orbach process is suggested as the predominant mechanism of magnetic relaxations in the high-temperature range, and the Arrhenius plots provide the effective relaxation energy barrier and pre-exponential factor of Ueff = 54.0 cm-1 and τ0 = 3.17 × 10-10 s, respectively. The direct estimation of the axial anisotropic parameter of 1 was successfully performed by high-field, multifrequency ESR measurements up to 55 T and 2.5 THz. The evaluated axial zero-field splitting (ZFS) energy of 57.0 cm-1 is comparable to the Ueff energy, confirming that the magnetic relaxations are initiated by the thermal excitation from the ground | M S⟩ = |±3/2⟩ states to the |±1/2⟩ states in the high-temperature range. The results of the ab initio calculations based on the CAS(7,5) SCF wave functions indicate that the ground states of 1 consist mainly of | M S⟩ = |±3/2⟩ states, while the first excited states are the mixture of | M S⟩ = |+1/2⟩ and |-1/2⟩.

Using food network unfolding to evaluate food-web complexity in terms of biodiversity: theory and applications.

Food-web complexity often hinders disentangling functionally relevant aspects of food-web structure and its relationships to biodiversity. Here, we present a theoretical framework to evaluate food-web complexity in terms of biodiversity. Food network unfolding is a theoretical method to transform a complex food web into a linear food chain based on ecosystem processes. Based on this method, we can define three biodiversity indices, horizontal diversity (DH ), vertical diversity (DV ) and range diversity (DR ), which are associated with the species diversity within each trophic level, diversity of trophic levels, and diversity in resource use, respectively. These indices are related to Shannon's diversity index (H'), where H' = DH  + DV  - DR . Application of the framework to three riverine macroinvertebrate communities revealed that D indices, calculated from biomass and stable isotope features, captured well the anthropogenic, seasonal, or other within-site changes in food-web structures that could not be captured with H' alone.

Improved Method for Isolation and Purification of Underivatized Amino Acids for Radiocarbon Analysis.

We have improved a method for isolation and purification of individual amino acids for compound-specific radiocarbon analysis (CSRA). To remove high-performance liquid chromatography (HPLC) eluent blanks from isolated amino acid fractions prior to the radiocarbon (Δ14C) measurement, each fraction was filtered through a membrane filter and then washed with diethyl ether twice. Radiocarbon measurements on standard amino acids processed and purified with the above method using elemental analyzer-accelerator mass spectrometry resulted in Δ14C values that were in strong agreement ( R2 = 0.998) with the original Δ14C value of each amino acid standard. From these measurements, we calculate dead and modern carbon contamination contributions as 1.2 ± 0.2 and 0.3 ± 0.1 µgC, respectively, which are consistent with direct assessments of HPLC procedural blanks of 1.0 ± 0.8 µgC per sample. These contamination constraints allow correction of measured Δ14C values for accurate and precise CSRA and are widely applicable to future archeological and biogeochemical studies.

Membrane rafts regulate sperm acrosome reaction via cAMP-dependent pathway in chickens (Gallus gallus domesticus).

Both transcriptionally and translationally inactive sperm need preassembled pathways into specific cellular compartments to function. Although initiation of the acrosome reaction (AR) involves several signaling pathways including protein kinase A (PKA) activation, how these are regulated remains poorly understood in avian sperm. Membrane rafts are specific membrane regions enriched in sterols and functional proteins and play important roles in diverse cellular processes, including signal transduction. Our recent studies on chicken sperm demonstrated that membrane rafts exist and play a role in multistage fertilization. These, combined with the functional importance of membrane rafts in mammalian sperm AR, prompted us to investigate the roles of membrane rafts in signaling pathways leading to AR in chicken sperm. Using 2-hydroxypropyl-ß-cyclodextrin (2-OHCD), we found that the disruption of membrane rafts inhibits PKA activity and AR without affecting protein tyrosine phosphorylation; however, these inhibitions were abolished in the presence of a cyclic 3,5-adenosine monophosphate (cAMP) analog. In addition, biochemical experiments showed a decrease in cAMP content in 2-OHCD-treated sperm, suggesting the involvement of soluble adenylyl cyclase (sAC) and transmembrane adenylyl cyclase (tmAC). Pharmacological experiments, combined with transcriptome analysis, showed that sAC and tmAC are present and involved in AR induction in chicken sperm. Furthermore, stimulation of both isoforms reversed the inhibition of PKA activity and AR in 2-OHCD-treated sperm. In conclusion, our results demonstrated that membrane rafts play an important role in AR induction by regulating the cAMP-dependent pathway and that they provide a mechanistic insight into membrane regulation of AR and sperm function in birds.

Synthesis of a Series of Monophthalocyaninato Cyclen Heavy Lanthanide(III) Complexes with Tetragonal Symmetry.

A series of the new monophthalocyaninato lanthanide complexes, [Ln(Pc)(cyclen)]Cl (Ln = Y, Tb, Dy, Ho, Er, Tm, and Yb; Pc = phthalocyaninato; cyclen = 1,4,7,10-tetraazacyclododecane) was synthesized and characterized. The crystallographic study of monophthalocyaninato complexes with a capping macrocyclic ligand having no π-conjugation while keeping the 4-fold symmetry is presented for the first time. All the six complexes were crystallized in a tetragonal structure of the I4mm space group. In addition to this, the Tb complex exhibited a triclinic crystal structure of the P1̅ space group. All of the complexes are isostructural, where both Pc and cyclen are coordinated to an Ln3+ ion giving an eight-coordinate square-antiprismatic geometry. The skew angle between the Pc and cyclen ligands was 45° in the tetragonal crystals and 41.5° in the triclinic crystal. The metal-nitrogen bond lengths were shorter on the Pc ligand side than cyclen side, presumably due to the difference in the charges of the ligands and the steric factor.

System Composed of Three Types of Electronic Angular Momenta: A J-S-L Triad in a Photoexcited π-Radical Bis(phthalocyaninato)terbium Single-Molecule Magnet.

Interactions of three different types of electronic angular momenta, namely, spin, orbital, and total angular momenta from different origins in a photoexcited neutral bis(phthalocyaninato)terbium single-molecule magnet (Pc2Tb, where Pc2- denotes a phthalocyaninato anion) have been studied. We have conducted varied-temperature and varied-magnetic-field magnetic circular dichroism (MCD) measurements on the highest occupied molecular orbital-lowest unoccupied molecular orbital electronic transition in the ligand side of the neutral Pc2Tb to reveal the quantum nature of the system composed of a π-radical with a spin angular momentum S, the 4f system with a total angular momentum J, and the cyclic π conjugate system with a photoinduced orbital angular momentum L. We have constructed a new theoretical model that gives a quantitative agreement to the temperature and magnetic field dependence of MCD. The theoretical analysis revealed that the system takes eight quantum states that can be expressed as | J z S z L z⟩ = |±±±⟩, where the three angular momenta are quantized along the fourfold symmetry axis. Thus, we have identified the existence of the magnetic interactions among the three angular momenta and quantitatively determined their magnitudes for the first time.

Modification of membrane cholesterol and desmosterol in chicken spermatozoa improves post-thaw survival and prevents impairment of sperm function after cryopreservation.

During cryopreservation, spermatozoa are subjected to cryodamage that leads to a decline in fertilisation ability. Due to the complex nature of this process, the initial trigger for cryodamage remains unknown. Recently, we demonstrated that cryopreservation induces early apoptotic changes characterised by phosphatidylserine (PS) translocation via sterol loss from the plasma membrane of chicken spermatozoa. This led us to hypothesise that sterol incorporation into membranes minimises cryodamage, thereby improving the quality of cryopreserved chicken spermatozoa. In the present study, treating spermatozoa with 1.5mgmL-1 cholesterol- and 3mgmL-1 desmosterol-loaded cyclodextrin (CLC and DLC respectively) increased post-thaw survival and motility. These effects appeared to be highly dependent the amount of sterol loaded into the spermatozoa. Localisation experiments confirmed the incorporation of exogenous cholesterol into the sperm head region. Detection of PS translocation showed that elevation of these sterols inhibited early apoptotic changes, thereby enhancing post-thaw survival. Furthermore, CLC and DLC treatment suppressed spontaneous acrosome reaction after cryopreservation, preserving the ability of spermatozoa to undergo acrosome reactions in response to physiological stimulation. These results demonstrate that loading sterols into chicken spermatozoa before cryopreservation enhances their quality by inhibiting early apoptotic changes and spontaneous acrosome reactions. The present study provides new mechanistic insight into cryodamage in chicken spermatozoa.

Selective Stabilization of the Spin States of a Magnetically Anisotropic Dysprosium Ion Induced by Photo-Excitation of the Associated Cyclic π-Conjugated System.

The presence of a new electronic interaction, which couples a 4 f-electronic system with a total angular momentum J and a photoexcited cyclic π-conjugated system with an orbital angular momentum L, in the bis(phthalocyaninato)dysprosium single-molecule magnet ([DyPc2 ]- ) is reported. Two π-π* excited states in the visible spectral region of the [DyPc2 ]- complex, which are denoted here as QL and QH , showed significantly different temperature and field dependences of the magnetic circular dichroism (MCD) A-term intensity. This phenomenon not only indicates the presence of a "J-L" interaction, but also that the interaction generates two different preferred orientations of the J-L pair, either parallel (for the QH band) or antiparallel (for the QL band), depending on the excitation energy. We have constructed a theoretical model that reproduces the temperature and field dependences, and quantitatively evaluated the J-L interaction.