Establishment of a new canine cell line (CCT) originated from a cutaneous malignant histiocytosis

A new canine cell line, named CCT, was established from the cutaneous malignant histiocytosis in a 4-year-old male Borzoi. CCT proliferated with loose adherence and doubling time was approximately 30 hr. When co-cultured with latex beads, CCT phagocytized beads vigorously. Lysozyme and vimentin were positive by immunostaining, and non-specific esterase and acid phosphatase were positive by cytochemical staining. These features indicated the cells had a histiocytic nature. Furthermore, by subcutaneous injection to nude mice CCT could successfully form tumors with the morphological and immunohistochemical features similar to the original tumor.     

Possible degradation of type I collagen in relation to yellowtail muscle softening during chilled storage

ABSTRACT:   In this paper, the detection of type I collagen degradation during the softening phenomenon of yellowtail muscle, was examined. Acid soluble collagen was isolated from dorsal ordinary muscle at death and after 24-h chilled storage. In the abundant ratio of subunit components, an increase in β12 chain (5.4 points) and a decrease in components with molecular weights larger than γ chain (7.0 points) after 24-h chilled storage, was found. Type I collagen was detected in the alkali-soluble fraction by SDS-PAGE. Its amount calculated from hydroxyproline contents in alkali-soluble fraction was increased from 0.097 mg/g muscle to 0.155 mg/g muscle during 24-h storage. The increased alkali-soluble collagen (0.058 mg/g muscle) was about 1.4% of whole collagen. These results suggest that a slight decomposition of type I collagen of yellowtail muscle may occur and subsequently becomes alkali-soluble corresponding to postmortem softening.     

Recycling of water for irrigation: Persistence of enteroviruses in sewage effluent and natural waters receiving the effluent

A virological analyses of a sewage treatment plant which provided chlorinated, activated sludge treated sewage effluent to irrigate a complete two-year crop of sugarcane was made. The raw, the activated sludge treated and the chlorinated sewage effluent, as well as streams and a harbor receiving sewage effluents were concentrated by either the polymer two-phase, PE-60, Al(OH)₃, protamine sulfate, or cellulose membrane method and assayed for human enteric viruses. Viruses were recovered from 100% (11/11) of the raw sewages tested at concentrations ranging from 27 to 19 000 PFU l^?1 while 76% (13/17) of the activated sludge treated effluent was positive at concentrations ranging from 7 to 5222 PFU l^?1. After chlorination, 58% (31/53) of the samples was positive for virus at concentrations ranging from 2 to 750 PFU l^?1. Human enteroviruses were also isolated from shallow flowing streams at distances up to 3 mi (5 km) from the closest known sewage effluent discharge point and from a harbor approximately 0.5 mi (0.8 km) from the point of sewage discharge entering the harbor. The viruses most often isolated were echovirus 7, coxsackievirus B-4, B-5 and poliovirus 1, 2, and 3. These results indicate that although activated sludge treatment plus chlorination remove approximately 90% of the virus from the raw sewage, the final treated sewage effluent, which is normally discharged into a stream and in this experimental study to irrigate sugarcane, still contains a significant concentration of infectious viruses. Furthermore, the recovery of enteroviruses from waterways at points distant from the sewage treatment plants indicates that sewage-borne viruses persist in natural water environment. The significance of enteric viruses in waters accessible to the public and used for irrigation purposes remains to be determined.     

Forest Floor Quality and N Transformations in a Temperate Forest Affected by Avian-Derived N Deposition

To evaluate the effects of avian-derived N deposition on forest C and N status, we investigated quantity and quality of litterfall, and chemical quality and N transformations in the forest floor from five sites that had been exposed to waterbird breeding colonies to differing degrees. The highest litterfall input of 2.6 t ha^?1 month^?1 was observed in the forest stand (Site 2) where intense bird breeding activity had been observed. Litterfall C/N was highest at the control site (no breeding activity) and decreased as the colonization stages advanced. Nitrogen concentration in litterfall and the forest floor was higher at those sites (Sites 3 and 4) where bird breeding had ceased and which represented the 'post-colony' situation, and consequently C/N and lignin/N of forest floor decreased. NH₄-N pool size in the forest floor was higher at Site 2, probably due to ongoing input and mineralization of bird excreta. Nitrification rate and percent nitrification were highest at Sites 3 and 4. From these results we conclude that study sites exposed to bird colonies show signs of N excess even after colonies are abandoned.     

Improved malonaldehyde assay using headspace solid-phase microextraction and its application to the measurement of the antioxidant activity of phytochemicals

A modified malonaldehyde (MA) assay for antioxidant activity, which involves derivatization and headspace solid-phase microextraction (HS-SPME) was developed and validated. The recovery of MA as 1-methylpyrazole (product of MA and N-methylhydrazine) from a headspace of an aqueous solution containing MA, buffer, surfactant, and cod liver oil using HS-SPME with a PDMS/DVB fiber was 91.3 +/- 3.38%. MA was analyzed by a gas chromatograph with a nitrogen-phosphorus detector, and its detection limit was 0.0103 nmol/mL. The antioxidant activities of natural compounds were determined as the percentage inhibition of MA formed from cod liver oil oxidized by Fenton's reagents in the above aqueous solution. Sesamol inhibited MA formation most (86.1%), followed by eugenol (84.4%), capsaicin (80.7%), ethylvanillin (45.3%), and vanillin (31.6%) at a level of 50 microg/mL. This method did not require any organic solvents and is a simple, fast, and a highly sensitive method for MA determination.     

Localization of the motoneurons innervating the hindlimb muscles in the spinal cord of the domestic fowl

The origins of the motor nerve fibers supplying the individual hindlimb muscles were elucidated in the fowl by the retrograde degeneration method. Chromatolytic cells were seen in the ipsilateral lamina 9 of the lumbosacral enlargement. Motoneurons innervating the individual hindlimb muscles were localized in the characteristic position of lamina 9. Both hip and thigh muscles are supplied by motoneurons of almost all the columns in lumbar segments and by those of dorsolateral columns in sacral segments, while the shank and foot muscles are supplied by motoneurons of ventromedial columns in sacral segments. The origins of the dorsal cord of the lumbosacral plexus are situated laterally in the lamina 9 and those of the ventral cord are located medially in it.     

A case of feline pancreatitis

A 14-year-old, spayed female, domestic shorthair cat was referred to us with anorexia, pyrexia, and jaundice. Total bilirubin (TBIL) and feline trypsin-like immunoreactivity (fTLI) levels were remarkably high. Based on laparoscopic biopsy of the pancreas, the cat was diagnosed as having pancreatitis. As a result of treatment with a synthetic protease inhibitor and corticosteroid, the TBIL and fTLI values returned to normal and the clinical course was good.     

Anthocyanin composition and antioxidant activity of the Crowberry (Empetrum nigrum) and other berries

The anthocyanin composition and antioxidant activity of the crowberry ( Empetrum nigrum) were studied. High-performance liquid chromatography (HPLC) combined with a diode array detector and electrospray ionization mass spectrometry were used for identification and quantification of individual anthocyanins. Freeze-dried crowberry powder was extracted with 80% methanol containing 0.5% acetic acid and subjected to HPLC. Thirteen kinds of anthocyanins were identified. The major anthocyanins were cyanidin-3-galactoside and delphinidin-3-galactoside, at 8.04 and 8.62 mg/g extract, respectively. The HPLC profile of crowberry extract was similar to bilberry and blueberry. The total content of anthocyanins in crowberry was 41.8 mg/g extract, higher than the other nine major berry species (2.5-38.8 mg/g extract). The antioxidant activity was also evaluated using the 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis(3-ethybenzothiazoline-6-sulfonic acid) radical quenching assays and the ferric reducing activity power assy. Crowberry extract exerted the strongest antioxidant activity. In conclusion, individual anthocyanins in crowberry were identified and then quantified in this study. Additionally, crowberry is suggested to be associated with a reduction in the risk of developing chronic diseases because of its strong antioxidant activity.     

Expression of alpha-hemoglobin stabilizing protein and cellular prion protein in a subclone of murine erythroleukemia cell line MEL

Alpha-Hemoglobin stabilizing protein (AHSP) functions as the erythroid-specific molecular chaperon for alpha-globin. AHSP gene expression has been reported to be downregulated in hematopoietic tissues of animals suffering from prion diseases though the mechanism remains to be clarified. Herein, we demonstrate that MELhipod8 cells, a subclone of murine erythroleukemia (MEL) cells, have prion protein (PrPc) on the cell surface and have highly inducible expression of the AHSP and alpha- and beta-globin genes, resembling the expression pattern of the PrP and AHSP genes in bipotential erythroid- and megakaryocyte-lineage cells followed by erythroid differentiation in normal erythropoiesis. Moreover, MELhipod8 cells exhibit greater effective erythroid differentiation with a population of hemoglobinized normoblast-like cells than that observed for the parental MEL cells. These findings suggest that MELhipod8 cells could provide a mechanism for downregulation of the AHSP gene in prion diseases.