Relationship between the responsiveness to multiple-ovulation treatment and the number of bovine oocytes collected by transvaginal follicle aspiration

To characterize factors affecting the number of bovine oocytes recovered transvaginally, a regression analysis was performed between the responsiveness to multiple-ovulation treatment and the number of oocytes recovered transvaginally. The number of embryos recovered following multiple-ovulation treatment and the number of oocytes recovered transvaginally increased when the number of follicles to be aspirated transvaginally increased (P<0.05. P<0.01). The number of cumulus-oocyte complexes recovered transvaginally also increased when the number of oocytes to be aspirated transvaginally increased (P<0.001). However, the number of viable embryos that recovered following multiple-ovulation treatment had no relation to the number of cumulus-oocyte complexes recovered transvaginally. These results suggested that more oocytes can be recovered from donors that have a high responsiveness to multiple-ovulation treatment.     

Apoptosis-independent Poor Morphology of Bovine Embryos Produced by Multiple Ovulation

In multiple ovulation and embryo transfer (MOET) programmes in cattle, a considerable number of morphologically poor-quality embryos continue to be produced; this is one of the limiting factors of the technique. Apoptosis has often been implicated in developmental arrest and fragmentation; these are regarded as poor traits of embryonic quality in mammalian pre-implantation embryos. In the present study, apoptosis was assessed in morphologically poor-quality embryos in comparison with good-quality embryos that were recovered from a MOET programme. Retarded embryos (two to 16 cell stage), morulae with severe fragmentation and morphologically good-quality morulae recovered from superstimulated cows at day 7 post-insemination were subjected to TdT-mediated dUTP nick-end labelling (TUNEL) and Hoechst staining. Cell nuclei that showed both TUNEL staining and apoptotic morphology were considered to be apoptotic. Apoptotic index (AI) was calculated as the percentage of apoptotic cells per embryo. Fifteen of 17 retarded embryos and 10 of 15 morphologically poor-quality morulae did not show signs of apoptosis. The mean AIs in the morphologically poor-quality embryos (two to 16 cell stage, 2.2%; poor morulae, 1.3%) were as low as that in the good-quality embryos (2.9%). These results suggest that another mode of developmental arrest and/or fragmentation that is independent of apoptosis occurs in morphologically poor-quality embryos recovered from MOET programmes.     

Concurrent West Nile virus and mycobacterium avium infection in a black-necked swan (Cygnus melanocoryphus).

An adult male black-necked swan (Cygnus melanocoryphus) was evaluated for a routine physical examination and West Nile virus vaccination. Nine days later, the swan was hospitalized for weakness, dehydration, tremors, and leukocytosis. Gradual clinical improvement was seen over a 2-wk course of supportive care. However, the animal became acutely weak and was found dead. Histopathology revealed severe granulomatous pneumonia and air sacculitis with dissemination to the liver, spleen, intestine, and coelom. Mycobacterial infection was confirmed by identification of acid-fast bacilli within tissue sections and by polymerase chain reaction. Concurrent West Nile virus infection was identified in sections of brain by using immunohistochemistry.     

猪隐秘杆菌型出血性坏死性脾炎

六月龄去势公猪表现昏睡、食欲不振、站立困难。尸检发现脾脏边缘呈多重出血灶。从脾脏、肾脏、肌肉和肝脏中分离出革兰氏阳性杆菌,对分离株(TO16177)的16S rDNA基因序列比较分析发现,其可能与未发表过的隐秘杆菌属HJ57-14E菌株(登记号:gi18873551)(比较675bp个碱基,相似性达99.7%)为同一个属。脾脏组织切片的组织学检查发现呈广泛性坏死和炎症,其中革兰氏阳性杆菌显而易见。肝脏中可见多病灶的坏死斑。免疫组织化学检测发现,分离株与抗化脓性隐秘杆菌属(Arcanobacterium pyogenes)和内氏放线菌(Actinomyces naeslundii)的多克隆抗体具有交叉反应,且与后者的交叉反应更强烈。相似的反应也见于扁桃体的化脓灶中分离株,偶尔也见于脾脏和淋巴结中的分离株。本研究结果表明,这种未公开发布的隐秘杆菌属的细菌会引起生长肥育猪多器官功能衰竭,而后继发急性出血性坏死性脾炎。     

Induction of cell proliferation in the rat liver by the short-term administration of ethyl tertiary-butyl ether

In the present study, in continuation of our previous experiment in order to investigate the mode of action (MOA) of ethyl tertiary-butyl ether (ETBE) hepatotumorigenicity in rats, we aimed to examine alterations in cell proliferation, that are induced by short-term administration of ETBE. F344 rats were administered ETBE at doses of 0, and 1,000 mg/kg body weight twice a day by gavage for 3, 10, 17 and 28 days. It was found that the previously observed significant increase of P450 total content and hydroxyl radical levels after 7 days of ETBE administration, and 8-OHdG formation at day 14, accompanied by accumulation of CYP2B1/2B2, CYP3A1/3A2, CYP2C6, CYP2E1 and CYP1A1 and downregulation of DNA oxoguanine glycosylase 1, was preceded by induction of cell proliferation at day 3. Furthermore, we observed an increase in regenerative cell proliferation as a result of ETBE treatment at day 28, followed by induction of cell cycle arrest and apoptosis by day 14. These results indicated that short-term administration of ETBE led to a significant early increase in cell proliferation activity associated with induction of oxidative stress, and to a regenerative cell proliferation as an adaptive response, which could contribute to the hepatotumorigenicity of ETBE in rats.     

Isolation and characterization of a low molecular weight peptide contained in sourdough

To investigate a sourdough-specific peptide, low molecular weight peptides were extracted from sourdough. The peptide fraction was subjected to two kinds of chromatography to separate the peptides. Reverse-phase chromatography of the peptide fraction in the sourdough showed certain specific peptides. The specific peptide fraction was further separated by gel filtration chromatography. Liquid chromatography tandem mass spectrometry analysis identified one of the peptides as VPFGVG (six-mer). This sequence was estimated to occur at the 287-292 position of a low molecular weight glutenin subunit. The peptide (designed as SDP1) was produced by proteases derived from wheat flour. SDP1 showed angiotensin-converting enzyme (ACE) inhibitory activity, and the 50% inhibitory peptide concentration (IC50) was 336 microM. It is possible that the SDP1 peptide partially confers ACE inhibitory activity in sourdough.     

Subcutaneous Soft Tissue Sarcoma with Rhabdoid Features in a Dog

A nine-year-old male beagle dog had a white spherical mass in the subcutis of the left lumbar region. Microscopically, spindle to oval cells diffusely proliferated in the fibrous and myxoid stroma. Many neoplastic cells showed rhabdoid features or vacuolated cytoplasm. Immunohistochemically, the neoplastic cells were positive for vimentin and S100 and partly positive for neuron-specific enolase and glial fibrillary acidic protein but were negative for von Willebrand factor, desmin and α-smooth muscle actin. Ultrastructurally, the neoplastic cells had abundant cytoplasmic processes and desmosome-like structures. Cytoplasmic inclusions of rhabdoid-featured cells in HE sections were composed of aggregates of intermediate filaments, and cytoplasmic vacuoles were identified as an invagination of cytoplasm. Although malignant peripheral nerve sheath tumor was suggested according to these results, the present case was diagnosed as a soft tissue sarcoma with rhabdoid features due to a lack of identification of the basal lamina under electron microscopy.     

An evaluation of one example of biotope roof in Japan: Plant development and invertebrate colonisation after 8 years

Biotope roofs in Japan are usually intensive green roofs that primarily include native plants and food plants for invertebrates and a pond and stones to create a wide range of habitats. The study aimed to evaluate the survivability of planted species, and colonisation by plants and invertebrates on a biotope roof and to suggest an appropriate planting design and maintenance scheme to optimize biodiversity benefits. An intensive green roof (150 m², substrate depth of 50 cm) was installed in 2002 on the ninth storey of a building at Chiba University, Japan. Twelve species of trees, 18 species of shrubs and 8 species of forbs (mainly native species) were planted and volcanic stones were used as mulch and to create habitats for invertebrates. No maintenance and no irrigation were applied for almost 8 years. A limited number of tree species, such as Myrica rubra and Cinnamomum camphora could grew well without maintenance and irrigation at a substrate depth of 50 cm. Overall, shrubs grew successfully and a high density planting seemed effective in increasing wind resistance. Most forbs disappeared, probably because of drought and competition with these colonising plants. Eleven plant species spontaneously colonised resulting in domination by Solidago altissima and Miscanthus sinensis. These two species were too aggressive, and selective weeding is required for species richness. In an invertebrate study, 46 species in 11 orders were observed and the highest number of invertebrate species was observed in the pond and shady areas.     

Revisiting the mechanism of <Emphasis Type="Italic">β-O</Emphasis>-4 bond cleavage during acidolysis of lignin IV: dependence of acidolysis reaction on the type of acid

The dependence of the acidolysis reaction of a C₆-C₃ dimeric nonphenolic β-O-4 type lignin model compound, 2-(2-methoxyphenoxy)-1-(3,4-dimethoxyphenyl) propane-1,3-diol (veratrylglycerol-β-guaiacyl ether, VG), on the type of acid applied was examined using three different acids [0.2 mol/l HCl, 0.2 mol/l HBr, and 0.1 mol/l (0.2 N) H₂SO₄ in 82% aqueous 1,4-dioxane at 85°C]. In the HCl system, the major reaction modes of the corresponding benzyl cation-type intermediate (BC), which is produced by protonation of the α-hydroxyl group of VG and successive release of the water molecule, are the abstraction of the β-proton and hydride transfer from the β-to the α-position. The liberation of formaldehyde from the γ-hydroxymethyl group of BC is the predominant reaction mode in the H₂SO₄ system. Apparently, an unknown reaction mode or modes is operative in the early stage of the HBr system that causes rapid disappearance of VG accompanied by the quantitative formation of 2-methoxyphenol without affording the common counterpart of a Hibbert’s ketone, 1-hydroxy-3-(3,4-dimethoxyphenyl) propan-2-one. The reaction mode in the HBr system changes with the progress of the reaction and is the same as that in the HCl system after the early stage.     

Axle-less F1-ATPase rotates in the correct direction

F1-adenosine triphosphatase (ATPase) is an ATP-driven rotary molecular motor in which the central gamma subunit rotates inside a cylinder made of three alpha and three beta subunits alternately arranged. The rotor shaft, an antiparallel alpha-helical coiled coil of the amino and carboxyl termini of the gamma subunit, deeply penetrates the central cavity of the stator cylinder. We truncated the shaft step by step until the remaining rotor head would be outside the cavity and simply sat on the concave entrance of the stator orifice. All truncation mutants rotated in the correct direction, implying torque generation, although the average rotary speeds were low and short mutants exhibited moments of irregular motion. Neither a fixed pivot nor a rigid axle was needed for rotation of F1-ATPase.