Improvement of the extraction procedure for hyaluronan from fish eyeball and the molecular characterization

The object of this study was to improve the isolation procedure of hyaluronan and to compare characteristics of hyaluronan from the eyeball of bigeye tuna Thunnus obesus with other sources. General sources of hyaluronan are from Streptococcus zooepidemicus and rooster comb. Hyaluronan can be also obtained from the vitreous of fish eyes. Pure hyaluronan of higher molecular weight was obtained by the following improved extraction procedure: the frozen vitreous of a tuna eyeball was used to avoid contamination with blood, muscle tissue, and other factors; extracting was carried out over a long time period under cold conditions; cetylpyridinium chloride was used in order to separate mucopolysaccharides containing hyaluronan in the initial procedure without the process of removing fat and protein by reagents. The hyaluronan obtained was characterized by gel permeation chromatography, dynamic light scattering measurements, and viscometry. The characteristics of hyaluronan from tuna eyeballs were similar to those from other sources. However, the viscosity was lower. The possible reason could be ascribed to the wide distribution of molecular size in the vitreous humor of fish eye.     

Effect of surimi processing on dimethylamine formation in fish meat during frozen storage

Trimethylamine-N-oxide (TMAO) degrades to dimethylamine (DMA) and formaldehyde, which cause a decline in fish quality. Suppression of DMA formation in frozen surimi was investigated using croaker, lizardfish, and walleye pollock. The leaching process in surimi processing was shown to reduce not only the TMAO, iron, and taurine content, but also to reduce unidentified factors that accelerate DMA formation in lizardfish muscle; in contrast, unidentified factors that suppress DMA formation were reduced in croaker and pollock muscle. Sucrose, used as a cryoprotectant, was shown to decrease DMA formation in vitro, likely due to the reduction in freezing-induced concentration of solutes. The effects of pH on DMA formation were different in minced frozen meat among the three species. DMA formation was not observed in croaker when the pH varied between 6 and 8. On the other hand, DMA was elevated in lizardfish under acidic conditions, and DMA formation in pollock was maximal when the pH of the meat was neutral. Thus, the suppression of TMAO degradation by surimi processing results from the removal of TMAO, iron, and reductants from fish meat; sucrose also reduces DMA formation. Furthermore, unidentified factors in croaker, lizardfish, and pollock meat substantially affect DMA formation.     

Cell corpse engulfment mediated by C. elegans phosphatidylserine receptor through CED-5 and CED-12

During apoptosis, phosphatidylserine, which is normally restricted to the inner leaflet of the plasma membrane, is exposed on the surface of apoptotic cells and has been suggested to act as an "eat-me" signal to trigger phagocytosis. It is unclear how phagocytes recognize phosphatidylserine. Recently, a putative phosphatidylserine receptor (PSR) was identified and proposed to mediate recognition of phosphatidylserine and phagocytosis. We report that psr-1, the Caenorhabditis elegans homolog of PSR, is important for cell corpse engulfment. In vitro PSR-1 binds preferentially phosphatidylserine or cells with exposed phosphatidylserine. In C. elegans, PSR-1 acts in the same cell corpse engulfment pathway mediated by intracellular signaling molecules CED-2 (homologous to the human CrkII protein), CED-5 (DOCK180), CED-10 (Rac GTPase), and CED-12 (ELMO), possibly through direct interaction with CED-5 and CED-12. Our findings suggest that PSR-1 is likely an upstream receptor for the signaling pathway containing CED-2, CED-5, CED-10, and CED-12 proteins and plays an important role in recognizing phosphatidylserine during phagocytosis.     

Isolation of monoclonal antibodies that inhibit the binding of infectious bursal disease virus to LSCC-BK3 cells

Three hybridoma cell lines producing monoclonal antibodies (MAbs) against LSCC-BK3 cells which are susceptible to infectious bursal disease virus (IBDV) infection were produced and characterized. The MAbs, designated T7, Q11 and Q13, inhibited the attachment of IBDV to LSCC-BK3 cells. Furthermore, these MAbs bound to LSCC-BK3 but not to nonpermissive cells in flow cytometry. MAb T7 detected a 110-kDa membrane protein of LSCC-BK3 cells, whereas Q11 and Q13 reacted with membrane proteins of molecular weights 58-, 85-, 90- and 110-kDa. These observations imply that the 110-kDa protein recognized by all the MAbs is associated with IBDV binding. The MAbs established in this study are useful for studying the interaction between IBDV and its target cell.     

Mast cell tumor in the nasal cavity of a dog

An 11-year-old male Shetland sheepdog displayed epistaxis and nasal discharge from the left nasal foramen. Cytological examination of a smear sample obtained by rhinotomy revealed neoplastic mast cells in the nasal cavity, a definitive diagnostic sign of mast cell tumor. The case was treated by surgery combined with radiotherapy and chemotherapy. Eighteen days after the last treatment, marked enlargement of the mandibular lymph nodes and facial edema developed, and the dog was euthanized at the owner's request. At necropsy, metastatic proliferation of mast cells was confirmed in the lymph nodes and liver, but no neoplastic mast cells were observed in the nasal cavity.     

Determination of GHG and ammonia emissions from stored dairy cattle slurry by using a floating dynamic chamber

We developed a system for measuring emissions from stored slurry by using a floating dynamic chamber. CH₄, CO₂, N₂O and NH₃ emitted from the storage tank of a dairy cattle farm in eastern Hokkaido were measured during summer 2008 (7/16–8/6), fall 2008 (10/2–10/26), spring 2009 (6/2–6/21) and winter 2009 (3/11). Average daily gas emission rates in summer, fall and spring were, respectively, 54.8, 54.2 and 34.3 g/m² for CH₄; 602, 274 and 254 g/m² for CO₂; 55.4, 68.2 and trace mg/m² for N₂O; and 0.55, 0.73 and 0.46 g/m² for NH₃. CH₄, CO₂ and NH₃ emission rates during the brief measurement period in winter were reduced to 1/4, 1/23 and 1/2, respectively, of summer emission rate levels. All gas emissions showed diurnal fluctuation and were greatest during the daytime, when the ambient temperature rose. CH₄, NH₃ and CO₂ emissions increased significantly during the daytime, and the daily emission (in grams) of each gas was positively correlated with maximum daily temperature. According to the combined spring, summer and fall measurements, the CH₄, N₂O and NH₃ annual emission factors were 1.42% (g CH₄/g volatile solids), 0.02% (g N₂O‐N/g total N) and 0.43% (g NH₃‐N/g total N), respectively.     

Anterior segment dysmorphogenesis of the eye and glaucoma in MG-W gerbils

Unilaterally swollen eyes were histopathologically characterized in four MG-W gerbils. The primary lesions resided in the anterior segment of the eye where neural crest cells play a critical role in embryonic development. They included indistinct filtration angle, unformed canal of Schlemm, hypoplastic iris, and ciliary body. The findings noted in the retina, optic nerve, optic tract, and lateral geniculate nucleus were consistent with the lesions induced following the persistent elevation of intraocular pressure as a result of insufficient drainage of aqueous humor. Thus, the present cases observed in the eyes of MG-W gerbils exemplified the anterior segment dysmorphogenesis associated with inadequate neural crest migration or differentiation, leading to subsequent glaucoma.     

Pyridinoline concentrations in muscular and skin collagen of fish and relationship between collagen solubility and pyridinoline concentration in fish muscular collagen

Pyridinoline (Pyr), one of the mature crosslinks of collagen, was determined in muscular collagen of three species of fish. The amounts of muscular Pyr in red sea bream, yellowtail, and tiger puffer were 3.4, 8.8, and 50.3 mmol/mol collagen, respectively, indicating that the Pyr concentration in muscular collagen differs greatly among fish species. The Pyr concentration in tiger puffer muscular collagen was the greatest, but it was only one-fourth that in rabbit muscle. As in mammalian skin collagen, Pyr was not detected in skin collagen of red sea bream and yellowtail. However, tiger puffer skin contained Pyr (3.75 mmol/mol collagen). The presence of Pyr would have a relationship to some features of tiger puffer skin, such as mechanical strength and thickness. Pyr concentrations in acid-soluble collagen (ASC), pepsin-solubilized collagen (PSC), and insoluble collagen (ISC) in muscles of three species of fish were determined. Pyr was found in ISC > PSC > ASC, from the highest to the lowest concentration, and the concentration in ISC was 45–200 times that in ASC. Therefore, Pyr crosslinks that are formed between collagen molecules would have a close relationship to collagen solubility.     

Genetic parameter estimation for number born alive at different parities in Landrace and Large White pigs

We estimated genetic parameters for number born alive (NBA) from the first to the seventh parities in Landrace and Large White pigs using three models. Analyzing 55,160 farrowing records for 12,677 Landrace dams and 43,839 for 10,405 Large White dams, we used a single‐trait animal model to estimate the heritability of NBA at each parity and a two‐trait animal model and a single‐trait random regression model to estimate the genetic correlations between parities. Heritability estimates of NBA at each parity ranged from 0.08 to 0.13 for Landrace and from 0.05 to 0.16 for Large White. Estimated genetic correlations between parities in all cases were positive. Genetic correlations between the first and second parities were slightly lower than those between other neighboring parities. Genetic correlations between more distant parities tended to be lower, in some cases <0.8. The results indicate the necessity to investigate the applicability of evaluating NBA at different parities as different traits (e.g., the first and later parities), although a repeatability model might still be reasonable.