Aberrant development of mammary glands, but precocious expression of beta-casein in transgenic females ubiquitously expressing whey acidic protein transgene

It has been suggested that whey acidic protein (WAP) may function as a protease inhibitor. However, the actual function of WAP remains obscure. We investigated the histological development of the mammary glands of transgenic mice ubiquitously expressing WAP and CAG/WAP transgene. Ubiquitous expression of WAP induced aberrant development of the lobular alveoli of the mammary glands: mammary alveoli that were either aberrantly large or small in size increased in number in the developing mammary glands of these transgenic females during pregnancy and lactation. The expression of beta-casein was precociously induced in the mammary glands of the transgenic females during early pregnancy and accompanying this was a histological observation that abnormally developed lobular alveoli filled with milk proteins appeared in the mammary glands of transgenic females during early pregnancy. However, during lactation, the development of mammary glands was impaired in transgenic females. To investigate the possible paracrine action of WAP associated with mammary gland aberration, we transplanted the mammary tissue of CAG/EGFP transgenic females into the fat pad of virgin CAG/WAP transgenic females and initiated pregnancy by mating. The development of mammary tissue transplanted to the recipient was histologically examined on day 3 of lactation. The results revealed that the development of grafted mammary tissues was impaired in a manner similar to that of the mammary glands of CAG/WAP transgenic females, indicating that the inhibitory effect of WAP acts via a paracrine mechanism. In vitro experiments using HC11 cells with forced expression of exogenous WAP demonstrated the inhibitory function of WAP on proliferation of mammary epithelial cells.     

Role of actin microfilaments in canine distemper virus replication in vero cells

Several studies have indicated that viruses require a specific cytoskeletal structure for replication in host cells. In this study, we examined the role of actin fiber in the replication of canine distemper virus (CDV), belonging to the Morbillivirus genus of the family Paramyxoviridae. For this purpose, we used two actin depolymerizing agents, cytochalasin-D (C-D) and mycalolide-B (ML-B). In Vero cells, C-D disrupted actin fibers distributed in the cytosol, but peripheral actin fibers remained intact. On the other hand, ML-B completely disrupted the actin fibers distributed in both areas. Treatment of Vero cells with C-D or ML-B inhibited the replication of CDV. Double staining of CDV-infected Vero cells with antibody to N-protein and rhodamine-phalloidin revealed the presence of N-protein in mid-cytoplasm. However, the N-protein was specifically localized at the submembrane region in the presence of C-D, whereas it was clustered in the presence of ML-B. Viral mRNA levels of N- and H-proteins were rather increased by treatment with C-D or ML-B. The treatment with ML-B strongly inhibited N-protein expression, whereas C-D only slightly inhibited N-protein expression. These results suggest that actin microfilaments distributed in the cytoplasm and on the membrane region in host cells may have a different role in the process of CDV replication.     

Antioxidant, anti-hyaluronidase and antifungal activities of Melaleuca leucadendron Linn. leaf oils

The present study examined the chemical composition, in vitro antioxidant, anti-hyaluronidase and antifungal activities of essential oils of Melaleuca leucadendron Linn. from Gundih-Central Java, Indonesia in different plant ages of 5, 10 and 15 years old. The Chemical composition of essential oils were analyzed by GC/MS. Twenty-six components were identified, of which 1,8-cineole (49.22–55.04 %), α-terpineol (8.79–10.70 %), d-limonene (5.58–6.39 %), and β-caryophyllene (5.03–7.64 %) were the main compounds in these oils. The antioxidant assay and anti-hyaluronidase assay showed that M. leucadendron leaf oils possess mild antioxidant activity with IC₅₀ between 7.21 and 9.23 mg/ml and anti-hyaluronidase activity with IC₅₀ between 1.94 and 3.03 mg/ml. The antifungal assay showed the effectiveness of these essential oils against Fomitopsis palustris (IC₅₀ 0.12–3.16 mg/ml), Trametes versicolor (IC₅₀ 0.01–0.06 mg/ml), Cladosporium cladosporioides (IC₅₀ 0.03–0.49 mg/ml), and Chaetomium globosum (IC₅₀ 0.06–0.15 mg/ml).     

Development of an air-injection press for preventing blowout of particleboard V: effects of board density and thickness on property of board manufactured from high-moisture particles

Particleboards with thickness of 10 mm and densities of 0.6, 0.7 and 0.8 g/cm³ were manufactured from high-moisture particles using urea–formaldehyde resin and the effectiveness of air injection was examined. The temperature in the 0.6 and 0.7 g/cm³ boards was lower with air injection than without during the initial to middle stages of pressing, while the temperature in the 0.8 g/cm³ board remained lower with air injection than without throughout the entire pressing process. Air injection reduced the pressing time required to manufacture the 0.6 and 0.7 g/cm³ boards and also increased the internal bond strength of boards of all densities. In the 0.6 and 0.7 g/cm³ boards, air injection reduced the modulus of rupture (MOR), while in the 0.8 g/cm³ boards, the MOR was similar between those manufactured by injecting and not injecting air. Air injection was also found to be effective for boards of high densities. The effectiveness of the air injection on thick boards was investigated by manufacturing 20-mm-thick boards of 0.7 g/cm³. Without air injection, it was not possible to manufacture the 20-mm-thick boards, even by extended hot pressing, but air injection allowed the boards to be manufactured by pressing for 16 min. Air injection was also shown to be effective for manufacturing thick boards.     

Performance of particleboard manufactured using air-injection press II: effects of board density and thickness on the performance of board manufactured from low-moisture particles

Particleboards of different densities (0.6, 0.7 and 0.8 g/cm³) and thicknesses (10 and 20 mm) were manufactured from low-moisture particles using an air-injection press. The effects of the air injection on preventing blowout of the boards of different densities and thicknesses were investigated by artificially creating blowout-prone conditions using metal frames. The effects of the air-injection pressure on the board performance were also investigated. 10-mm-thick boards of 0.8 g/cm³ pressed at 170 °C blew out when air was not injected, but were successfully manufactured by injecting air. 10-mm-thick boards at 150 °C showed constant internal bond (IB), regardless of density, but at 170 °C, IB was higher in boards of higher densities. This was likely due to accelerated hardening of the urea–formaldehyde resin at 170 than 150 °C. At both pressing temperatures, low air-injection pressure did not cause blowout and a reduction in board performance. Air injection also prevented the blowout of thick boards of 20 mm and enabled successful manufacture, showing its effectiveness. The IB of the 20-mm-thick board manufactured using the air-injection press exceeded that of 20-mm-thick board manufactured using an ordinary hot press.     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

Development of biosensor for measuring oxidative stress of fish

Fisheries Science - To elucidate the dynamics of oxidative stress in fish, it is necessary to know the concentration of superoxide anions as a precursor to various reactive oxygen species in the...     

First application of PCR-Luminex system for molecular diagnosis of fungicide resistance and species identification of fungal pathogens

Fungicide resistance in plant pathogens is often caused by a single point mutation in a gene encoding fungicide target proteins. Such is the case for resistance to MBI-D (inhibitors of scytalone dehydratase in melanin biosynthesis) fungicides in rice blast fungus (Magnaporthe oryzae), which is caused by a mutation in the scytalone dehydratase gene that results in a replacement of valine with methionine at codon 75 of the fungicide target protein. PCR-Luminex, a novel system developed for high-throughput analysis of single nucleotide polymorphisms (SNPs) was successfully introduced to diagnose MBI-D resistance using specific oligonucleotide probes coupled with fluorescent beads. The PCR-Luminex system was further tested for its potential in identifying species causing Fusarium head blight on wheat. Four major pathogens, Fusarium graminearum (=F. asiaticum), F. culmorum, F. avenaceum, and Microdochium nivale, known to cause the disease, were tested, and the species were identified using the PCR-Luminex method. So far, this report is the first on the application of the DNA-based PCR-Luminex system in the area of crop protection and/or agricultural sciences.     

Genetic analysis and PCR-based identification of major <Emphasis Type="Italic">Fusarium</Emphasis> species causing head blight on wheat in Japan

Identifying the Fusarium species cause Fusarium head blight (FHB) and produces mycotoxins in wheat and other cereal is difficult and time consuming because of confusing phenotypic classification systems. In Japan, the F. graminearum complex, F. culmorum, F. avenaceum, and Microdochium nivale predominantly cause FHB. The internal transcribed spacer (ITS) and 5.8S of rDNA, a partial sequence of β-tubulin and mitochondrial cytochrome b (cytb) genes of the four species were PCR-amplified and analyzed. On the basis of the ITS, β-tubulin and cytb sequences, F. avenaceum and M. nivale are distinct from the F. graminearum complex and F. culmorum, whereas the F. graminearum complex is closely related to F. culmorum. Moreover, thiophanate–methyl-resistant isolates of the F. graminearum complex and F. culmorum did not have an amino acid substitution at amino acid codon 198 or 200 of β-tubulin. In contrast, very highly or highly thiophanate–methyl-resistant isolates of M. nivale had Glu (GAG) substituted with Ala (GCG) or Lys (AAG) at codon 198, respectively. The allele-specific PCR assay was used to identify the F. graminearum complex and F. culmorum, and these Fusarium species could be distinguished rapidly.