The luteotrophic function of galectin-1 by binding to the glycans on vascular endothelial growth factor receptor-2 in bovine luteal cells

The corpus luteum (CL) is a temporary endocrine gland producing a large amount of progesterone, which is essential for the establishment and maintenance of pregnancy. Galectin-1 is a β-galactose-binding protein that can modify functions of membrane glycoproteins and is expressed in the CL of mice and women. However, the physiological role of galectin-1 in the CL is unclear. In the present study, we investigated the expression and localization of galectin-1 in the bovine CL and the effect of galectin-1 on cultured luteal steroidogenic cells (LSCs) with special reference to its binding to the glycans on vascular endothelial growth factor receptor-2 (VEGFR-2). Galectin-1 protein was highly expressed at the mid and late luteal stages in the membrane fraction of bovine CL tissue and was localized to the surface of LSCs in a carbohydrate-dependent manner. Galectin-1 increased the viability in cultured LSCs. However, the viability of LSCs was decreased by addition of β-lactose, a competitive carbohydrate inhibitor of galectin-1 binding activity. VEGFR-2 protein, like galectin-1, is also highly expressed in the mid CL, and it was modified by multi-antennary glycans, which can be recognized by galectin-1. An overlay assay using biotinylated galectin-1 revealed that galectin-1 directly binds to asparagine-linked glycans (N-glycans) on VEGFR-2. Enhancement of LSC viability by galectin-1 was suppressed by a selective inhibitor of VEGFR-2. The overall findings suggest that galectin-1 plays a role as a survival factor in the bovine CL, possibly by binding to N-glycans on VEGFR-2.     

Nitrogen promotes water consumption in seedlings of Cryptomeria japonica but not in Chamaecyparis obtusa

To evaluate the influence of excessive N deposition on the water consumption of a Japanese plantation forest, 1-year-old seedlings of major plantation trees, Cryptomeria japonica (Japanese cedar) and Chamaecyparis obtusa (Hinoki cypress), were treated with combinations of two N levels (Moderate N and High N) and two soil water conditions (Dry and Moist) for 4 months. The High N treatment received five times as much N as in the Moderate N treatment; the total amount of N added in the High N treatment was roughly 25 times the annual N deposition in precipitation. An increase in soil N availability increased the needle transpiration rate, needle biomass, and needle N content of C. japonica under the Moist treatment, whereas those of C. obtusa were not significantly affected by soil N treatment at either soil water level. Needle N content in C. japonica was positively related to needle photosynthetic rate and transpiration rate. Our results suggested that excessive N deposition has the potential to enhance water consumption in C. japonica stands on moist soils. However, the effects of increased N deposition would be insignificant for C. japonica grown on dry sites. Unlike in C. japonica, water consumption in C. obtusa would be unlikely to respond to excess N deposition, regardless of the soil moisture level. Moreover, the significant reduction in the fine root to needle ratio observed with excessive N application in C. japonica under both Dry and Moist treatments suggests that excessive N deposition is likely to cancel out the tree's morphological adaptation to drought.     

Abnormal clonalities of B-lymphocytes in bovine leukemia virus-infected cattle with persistent lymphocytosis

Peripheral B-lymphocyte clonality of 274 bovine leukemia virus-infected cattle with lymphocytosis was analyzed using clonality PCR based on sequences of the variable region of the bovine immunoglobulin H chain. None of the cattle showed monoclonal proliferation, while 10, 31, and 233 showed minor-clonal, oligoclonal, and polyclonal proliferation, respectively. A total of 163 cattle were analyzable the following year, and lymphocytosis was maintained in 157, indicating persistent lymphocytosis (PL). B-lymphocyte clonality of the 157 PL cattle was minor-clonal in 6 (3.8%), oligoclonal in 8 (5.1%), and polyclonal in 143 (91.1%). A higher rate of enzootic bovine leukosis (EBL) onset within a year was observed in PL cattle with minor-clonal (50.0% (3/6)) and oligoclonal (25.0% (2/8)) proliferation compared to those with polyclonal (5.6% (8/143)) proliferation. Minor-clonal and oligoclonal proliferation in PL cattle may be a prognosis factor for developing EBL.     

Epidemiological study of Kabuto Mountain virus,a novel uukuvirus,in Japan

Kabuto Mountain virus (KAMV), the new member of the genus Uukuvirus, was isolated from the tick Haemaphysalis flava in 2018 in Japan. To date, there is no information on KAMV infection in human and animals. Therefore, serological surveillance of the infection among humans and wild mammals was conducted by virus-neutralization (VN) test and indirect immunofluorescence assay (IFA). Sera of 24 humans, 59 monkeys, 171 wild boars, 233 Sika deer, 7 bears, and 27 nutria in Yamaguchi Prefecture were analyzed by VN test. The positive ratio of humans, monkeys, wild boars, and Sika deer were 20.8%, 3.4%, 33.9% and 4.7%, respectively. No positive samples were detected in bears and nutria. The correlation coefficients between VN test and IFA in human, monkey, wild boar, and Sika deer sera were 0.5745, 0.7198, 0.9967 and 0.9525, respectively. In addition, KAMV was detected in one pool of Haemaphysalis formosensis ticks in Wakayama Prefecture. These results indicated that KAMV or KAMV-like virus is circulating among many wildlife and ticks, and that this virus incidentally infects humans.     

Mixed Thymoma in a Young Cynomolgus Monkey (Macaca fascicularis)

A mass with a diameter of 1.5 cm was found in the thymus of a 4-year and 3-month-old male cynomolgus monkey. Microscopically, the mass consisted of two different patterns of proliferation, dense or fascicular proliferation of elongated spindle cells in a sporadic storiform pattern and dense proliferation of thymic cortex-like lymphoid cells in which the multifocal pale nests resembling the thymic medulla were distributed. In these pale nests, large dendriform cells sometimes forming Hassall's corpuscles were present. The proliferating spindle cells were positive for cytokeratin. The lymphoid cells in the mass were positive for CD3. We concluded that the mass consisted of the neoplastic thymic epithelium with thymocytes proliferation containing medullary differentiation. The mass was diagnosed as a mixed thymoma according to the WHO classification of thymomas in humans. Mixed thymoma is characterized as a mixture of two types of proliferative lesions, spindle-shaped epithelial proliferation and a lymphocyte predominant lesion with or without medullary differentiation. To the best of our knowledge, this is the first report concerning thymoma in monkeys.     

Silkworm recombinant bovine zona pellucida protein 4 (bZP4) as a potential female immunocontraceptive antigen; impaired sperm-oocyte interaction and ovarian dysfunction

Porcine zona pellucida proteins (ZPs) have been utilized as female immunocontraceptive antigens. The purpose of this study was to explore the potential use of silkworm recombinant bovine ZP4 as an alternative. When the protein was injected with monophosphoryl lipid A (MPL) – an immuno-stimulative agent – into two female goats, marked elevation of the anti-ZP4 titer was detected. Application of the purified specific IgG to a porcine in vitro fertilization system reduced the sperm penetration rate. In one goat, the cyclic profile of serum progesterone disappeared as the anti-ZP4 titer increased. Histological examination of the ovaries revealed degeneration of antral follicles with sparse infiltration of inflammatory cells in the theca, indicating that autoimmune oophoritis had been induced. Together, the present results suggest that recombinant ZP4 disturbs fertilization and exerts a pathogenic effect on follicle development in goats, thus indicating its potential as a female immunocontraceptive antigen.     

Comparison of Physical Properties of Wheat Starch Gels with Different Amylose Content

The effects of amylose content and other starch properties on concentrated starch gel properties were evaluated using 10 wheat cultivars with different amylose content. Starches were isolated from grains of two waxy and eight nonwaxy wheat lines. The amylose content of waxy wheat lines was 1.4–1.7% and that of nonwaxy lines was 18.5–28.6%. Starch gels were prepared from a concentrated starch suspension (30 and 40%). Gelatinized starch was cooled and stored at 5°C for 1, 8, 16, 24, and 48 hr. The rheological properties of starch gels were studied by measuring dynamic viscoelasticity with parallel plate geometry. The low‐amylose starch showed a significantly lower storage shear modulus (G′) than starches with higher amylose content during storage. Waxy starch gel had a higher frequency dependence of G′ and properties clearly different from nonwaxy starches. In 40% starch gels, the starch with lower amylose showed a faster increase in G′ during 48 hr of storage, and waxy starch showed an extremely steep increase in G′. The amylose content and concentration of starch suspension markedly affected starch gel properties.     

Porencephaly in a cynomolgus monkey ( macaca fascicularis )

Porencephaly was observed in a female cynomolgus monkey (Macaca fascicularis) aged 5 years and 7 months. The cerebral hemisphere exhibited diffuse brownish excavation with partial defects of the full thickness of the hemispheric wall, and it constituted open channels between the lateral ventricular system and arachnoid space. In addition, the bilateral occipital lobe was slightly atrophied. Histopathologically, fibrous gliosis was spread out around the excavation area and its periphery. In the roof tissue over the cavity, small round cells were arranged in the laminae. They seemed to be neural or glial precursor cells because they were positive for Musashi 1 and negative for NeuN and GFAP. In the area of fibrous gliosis, hemosiderin or lipofuscin were deposited in the macrophages, and activated astroglias were observed extensively around the excavation area.     

Histopathology of incidental findings in cynomolgus monkeys ( macaca fascicularis ) used in toxicity studies

The purpose of our publication is to widely communicate pictures of spontaneous findings occurring in cynomolgus monkeys. Focal lymphoplasmacytic infiltration is commonly seen in the general organs. The frequency and severity of these lesions may be influenced by the administration of drugs with an effect on the immune system. Lymphoplasmacytic infiltration in the lamina propria of the stomach is also frequently seen in cynomolgus monkeys, and it is caused mainly by a Helicobacter pylori infection. Various degrees of brown pigments are observed in various organs, and it is possible to distinguish the material of the pigments by its morphological features and site. A focal/segmental glomerular lesion is occasionally seen in a section of the kidney, and the minimal lesion has no influence on the urinalysis. We showed the common glomerular lesions in HE-stained sections, as well as in PAM- or PAS-stained sections, for understanding the details. Young and pubertal monkeys are usually used in toxicity studies; therefore, understanding various maturation stages of the genital system is important. In particular, the female genital system needs to be understood in the morphology, because their cyclic changes are different from other laboratory animals. Thus, we present the normal features of the cyclic changes of the female genital organs. Furthermore, we provide more information on spontaneous findings in cynomolgus monkeys for exact diagnoses in toxicity studies.     

ELISAs based on rNcGRA7 and rNcSAG1 antigens as an indicator of Neospora caninum activation

Bovine abortion caused by the Apicomplexa parasite Neospora caninum is a major economical problem in the livestock industry worldwide. However, the relationship between N. caninum infection and abortion is still unknown. Our study aimed to elucidate the relationship between parasite-specific antibody responses, parasite stages and abortion. In experimentally infected cattle, anti-NcGRA7 IgG1 antibody was predominantly detected during the acute infection stage, while the production of anti-NcSAG1 IgG1 antibody was observed during both acute and chronic stages. Furthermore, levels of anti-NcGRA7 IgG2 antibody were lower than those of anti-NcSAG1 IgG2 antibody. When tested on cattle with Neospora-associated abortion, positive rates of the anti-NcGRA7 IgG2 antibody were significantly lower than those of the anti-NcSAG1 antibody, although there was no difference in IgG1 antibody-positive rates between the two antigens. In addition, anti-NcGRA7 IgG2 antibodies were not detected in cattle for more than 30days after abortion. Our results suggested that anti-NcGRA7 and anti-NcSAG1 antibodies are suitable indicators for the activation stage of N. caninum infection and broad detection of the infection, respectively. In conclusion, the use of recombinant NcGRA7 and NcSAG1-based ELISAs will be useful for evaluating the abortion risk associated with N. caninum infection.