Physicochemical properties of akabane virus: A member of the simbu arbovirus group of the family bunyaviridae

The multiplication of Akabane virus was not inhibited in the presence of 5-iodo-2′-deoxyuridine, indicating the presence of RNA. The virus was considered to have an envelope, as it was sensitive to ether and chloroform. It was readily inactivated by deoxycholate and trypsin, but was not precipitated by protamine sulphate. The virus was very labile at pH 3 and also rather heat-labile. Akabane virus was readily filtered through membrane filters of 200 or 100-nm pore size, but not through 50-nm filters. Equilibrium centrifugation in a CsCl density gradient gave a peak of infectivity and hemagglutinin at a density of 1.22 g/ml. The peak fractions thus obtained contained numerous virus particles, roughly spherical, variable in size, 70 to 130 nm in diameter, and mostly having a ragged, closely adherent envelope with projections, when examined, following phosphotungstic acid negative staining, in an electron microscope.     

Shifts in disease resistance induced by growth regulators

Experiments on the effect of various compounds with plant growth regulating activity led to the hypothesis that conditions inhibitory to indoleacetic acid (IAA) action or leading to a decrease in the IAA level in cucumber seedlings would be unfavourable for the development of cucumber scab, caused byCladosporium cucumerinum. Susceptibility decreased as the result of treatment with growth retardants, which cause an increase in IAA-oxidase activity in the plants, whereas application of indoleacetic acid or compounds expected to decrease the rate of oxidative breakdown of IAA in the seedlings resulted in an increase of susceptibility. Growing the plants under various periods of illumination also influenced both the susceptibility of the plants and the IAA-oxidase activity in the hypocotyl tissue.To obtain further information about the relation between the increase in resistance and the increase in the rate of IAA oxidation, the effect of one compound, viz. L-threo-β-phenylserine (abbr. phenylserine) was studied in more detail. The rate of oxidative breakdown of IAA was increased in extracts of hypocotyls and cotyledons of plants treated with phenylserine as compared with extracts of tissue of untreated plants. Extracts of phenylserine-treated plants were found to contain a higher cofactor and/or a lower inhibitor content than those of control plants. The substances involved were not identified, but this result may indicate a shift in the concentrations of various phenols in the tissue.The question must remain open whether a shift in the concentration of phenols some-how effected by phenylserine treatment determines both degree of susceptibility and rate of IAA oxidation independently, or whether the rate of IAA breakdown and the resulting change in IAA content in the plants is directly related to the degree of susceptibility.     

Matrix-metalloproteinases-2 and -9 production in bovine endometrial cell culture

In vitro cell culture is a convenient tool for studying cellular mechanisms. In the present study, production of matrix-metalloproteinases (MMPs) in bovine endometrial (containing both epithelial and stromal cells) monolayer cells was examined. Blastocysts attached to the endometrial cells in a monolayer culture were examined for their effects on MMP-2 production. Initial attachment of blastocysts to the monolayer inhibited MMP-2 production by endometrial cells. But once trophoblast cells began to migrate into the endometrial cell layer, MMP-2 production increased, and at the same time MMP-9 production also became evident in the medium. In order to understand how blastocysts affected MMP-2 production, we examined the effect of progesterone, estradiol, insulin-like growth factors (IGFs), tumor necrosis factors (TNFs), and interferon-tau (IFN-tau) supplementation. It was IFN-tau that inhibited the production of MMP-2. In addition, progesterone at a lower dose appeared to inhibit MMP-2 production. Both TNF-alpha and TNF-beta strongly stimulated the production of MMP-2 and MMP-9, whereas IGFs had no effect. Based on these findings, it appears that conceptus has the capacity to inhibit MMP activity.     

Matrix metalloproteinases-2 and -9 activities in bovine follicular fluid of different-sized follicles: relationship to intra-follicular inhibin and steroid concentrations

Matrix metalloproteinases (MMPs) play very important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, ovulation and atresia. The aim of the present study was to determine the content of gelatinases in follicular fluid in various sized bovine follicles. Bovine ovaries were collected from local slaughterhouse and follicular fluid from follicles of 2 to over 25 mm in diameter was collected. Gelatinase activity within the follicular fluid was analyzed by gelatin zymography. The concentration of inhibin in the follicular fluid was also measured by immunoblot analysis. The proMMP-2 and alpha-subunit (alphaN) inhibin was detected in all follicles regardless of their size. The abundance of proMMP-2 varied with follicular size, while alphaN inhibin increased significantly (P<0.01) in follicles of 10-14 and 15-20 mm in size. There was a positive and negative correlation between estradiol (E(2)) and progesterone (P(4)) concentrations with abundance of proMMP-2, respectively. Follicles of diameter over 25 mm had greater proMMP-9 activity than other follicles. These same follicles had significantly (P<0.01) lower inhibin levels than follicles of 10-14 and 15-20 mm in size. In conclusion, these results suggest a significant role of these proteases in growth and development of bovine follicle, particularly proMMP-2 and active MMP-2 activities in the follicular fluid could serve as markers of follicular health while abundance of proMMP-9 may possibly denote a follicular cyst.     

Concentrations and specific antibodies to staphylococcal enterotoxin-C and toxic shock syndrome toxin-1 in bovine mammary gland secretions,and inflammatory response to the intramammary inoculation of these toxins

To investigate the pathological role of staphylococcal enterotoxins (SEs) and toxic shock syndrome toxin-1 (TSST-1) in bovine mastitis, the production of SEs and TSST-1 was investigated in staphylococci isolated from 120 mammary gland secretions (MGS, 51 from no clinical sign-mammary glands and 69 from staphylococcal mastitic-mammary glands) collected from dairy farms where staphylococcal mastitis frequently occurred in Miyagi and Yamagata prefectures from 1997 to 1998. Concentrations of these toxins and specific antibody titers in each MGS were also measured. Furthermore, SEC and TSST-1 were inoculated into lactating mammary glands and inflammatory responses were analyzed. A high percentage of staphylococci including Staphylococcus aureus and coagulase-negative staphylococci isolated from both no clinical sign- and mastitic-MGS produced both SEC and/or TSST-1. The concentration of SEC increased with the severity of the mastitis, and was significantly higher (P<0.05) in acute mastitic-than in no clinical signs-MGS. Titers of specific antibodies to TSST-1 in MGS were significantly higher (P<0.05) than those to SEC, regardless of whether or not the cows were lactating or mastitic. Specific antibodies purified from MGS neutralized each toxin in vitro. A significant increase (P < 0.05) in somatic cell counts was induced by the intramammary inoculation of SEC but not TSST-1. These findings indicated that SEC rather than TSST-1 plays an important role in the pathology of staphylococcal bovine mastitis. The inflammatory activity of TSST-1 was probably neutralized by specific antibodies in MGS.     

Bronchoalveolar lavage fluid findings in the atelectatic regions of anesthetized horses

To evaluate the effects of compression atelectasis on the composition of pulmonary secretions in anesthetized horses, cytological and biochemical examinations were performed on bronchoalveolar lavage (BAL) fluids obtained from both dependent and independent lung regions. Six horses were anesthetized with isoflurane and oxygen for 120 min, and were positioned in right lateral recumbency. Percentage of neutrophils and total protein concentration in BAL fluids significantly increased at the end of anesthesia, and total phosphorous concentration significantly decreased at 72 hr after anesthesia in dependent lung. Such changes were not observed within 168 hr after anesthesia. These findings suggest that the effects of compression atelectasis on the composition of pulmonary secretions may be eliminated within 168 hr after anesthesia.     

Survey on wild rodents for endoparasites in Iwate Prefecture, Japan

Wild rodents (58 Apodemus speciosus, 29 A. argenteus and 7 Microtus montebelli) were surveyed for endoparasites in Iwate Prefecture, Japan, from October to December 1995 and from April to October 1996. Two trematodes (Echinostoma macrorchis, Plagiorchis muris), 4 or more cestodes (Hymenolepis diminuta, Raillietina coreensis, Cladothyridium spp., Cysticercus fasciolaris), 12 nematodes (Carolinensis minutus, Eucoleus sp., Heligmosomoides kurilensis, H. protobullosus, H. speciosus, Heterakis spumosa, Rhabditis (Pelodera) orbitalis, Rictularia cristata, Syphacia emileromani, S. frederici, S. montana, Trichuris sp.) and 3 protozoans (Giardia sp., Trichomonas sp., Trypanosoma sp.) were identified. The two species of Apodemus were similar to each other, but they were extremely different from M. montebelli in parasite fauna.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518