Development and biological properties of a new live attenuated mumps vaccine

To develop a new live attenuated mumps vaccine, a wild mumps Y7 strain isolated from a patient who developed mild parotitis was treated with nitrosoguanidine and ultraviolet, followed by selection of a temperature-sensitive clone. The selected clone, Y125, showed stable temperature-sensitivity in Vero cells. Intraspinal inoculation of marmosets with the Y125 produced only minimal histopathological changes, while intracerebral inoculation of neonatal rats revealed that the Y125 did not cause hydrocephalus. Both these effects of the Y125 were similar to those of the non-neurovirulent Jeryl Lynn strain. Furthermore, subcutaneous inoculation of the Y125 induced high levels of neutralizing antibodies in all Cercopithecus monkeys examined. Although the safety and immunogenicity should be confirmed in further field trials in humans, the present results indicate that the Y125 could be a promising vaccine candidate.     

Comparison of the glucosinolate-myrosinase systems among daikon (Raphanus sativus, Japanese white radish) varieties

Myrosinase is a cytosolic plant enzyme present in daikon ( Raphanus sativus, Japanese white radish) roots that hydrolyzes 4-methylthio-3-butenyl glucosinolate (MTBGLS) into the natural pungent agent 4-methylthio-3-butenyl isothiocyanate (MTBITC), which possesses antimicrobial, antimutagenic, and anticarcinogenic properties. The concentration of MTBGLS, myrosinase activity, and production of MTBITC in seven daikon varieties (one conventional and six heirlooms) were determined to rank the activity of the glucosinolate-myrosinase system and identify critical factors influencing the production of MTBITC. The six heirloom varieties produced 2.0-11.5 times higher levels of MTBITC as compared to the conventional variety, Aokubi, which is consumed by the present Japanese population. The myrosinase was located exclusively in the outer epidermal layer in Aokubi, and MTBGLS was widely distributed throughout the root tissue. Although the skin is a potentially rich source of myrosinase in Aokubi, the skin is usually peeled off in the current practice of preparing daikon for cooking. New practices are therefore proposed for the preparation of daikon tubers that eliminate the peeling of the skin to avoid removing the enzyme needed to convert MTBGLS to the health-beneficial MTBITC. It is also concluded that the consumption of heirloom daikon varieties in addition to changes in food preparation will optimize the health benefits of daikon.     

Analysis of phenolic compounds in white rice, brown rice, and germinated brown rice

Two hydroxycinnamate sucrose esters, 6'-O-(E)-feruloylsucrose and 6'-O-(E)-sinapoylsucrose, were isolated from methanol extracts of rice bran. Soluble and insoluble phenolic compounds as well as 6'-O-(E)-feruloylsucrose and 6'-O-(E)-sinapoylsucrose from white rice, brown rice, and germinated brown rice were analyzed using HPLC. The results demonstrated that the content of insoluble phenolic compounds was significantly higher than that of soluble phenolics in rice, whereas almost all compounds identified in germinated brown rice and brown rice were more abundant than those in white rice. 6'-O-(E)-Feruloylsucrose (1.09 mg/100 g of flour) and 6'-O-(E)-sinapoylsucrose (0.41 mg/100 g of flour) were found to be the major soluble phenolic compounds in brown rice. During germination, an approximately 70% decrease was observed in the content of the two hydroxycinnamate sucrose esters, whereas free phenolic acid content increased significantly; the ferulic acid content of brown rice (0.32 mg/100 g of flour) increased to 0.48 mg/100 g of flour and became the most abundant phenolic compound in germinated brown rice. The content of sinapinic acid increased to 0.21 mg/100 g of flour, which is nearly 10 times as much as that in brown rice (0.02 mg/100 g of flour). In addition, the total content of insoluble phenolic compounds increased from 18.47 mg/100 g of flour in brown rice to 24.78 mg/100 g of flour in germinated brown rice. These data suggest that appropriate germination of brown rice may be a method to improve health-related benefits.     

Cerebellar focal granulomatous meningoencephalitis in a dog: clinical findings and MR imaging

The authors encountered a dachshund dog, presenting vestibular disorder. On magnetic resonance (MR) imaging, a mass showing isointensity on the T1- and T2-weighted images and enhanced by contrast medium, was observed in the right cerebellum. In addition, the periphery of the mass showed isointensity on the T1-weighted image and hyperintensity on the T2-weighted image, suggesting sever oedema. Although the dog underwent surgery, it died. The mass was diagnosed pathologically as a granulomatous meningoencephalitis.     

Peptides derived from dolicholin, a phaseolin-like protein in country beans (Dolichos lablab), potently stimulate cholecystokinin secretion from enteroendocrine STC-1 cells

Peptides derived from soybean beta-conglycinin and pork protein stimulate cholecystokinin (CCK) secretion from the enteroendocrine cells (EECs) and suppress food intake. Here we examined CCK-releasing activities from the enteroendocrine cell line STC-1, in peptides derived from underutilized legumes, and found much higher activity in the peptic hydrolysate of Country beans (CBP) compared to that from other legume-derived peptides including beta-conglycinin peptone. Active components in CBP were separated into acetonitrile-soluble fractions, but the activities were abolished after pronase treatment. To identify the Country bean protein containing the active peptides, Country bean protein extracts in an alkaline solution (CBE) were fractionated based on isoelectric point or molecular weight. Peptones prepared from CBE fractions containing a 51 kDa major protein stimulated CCK release, but other fractions did not. N-Terminal sequence analysis indicated that the 51 kDa protein is a phaseolin-like globular protein, and we designated this protein dolicholin. These results indicate that Country bean-derived peptides are very potent legume peptides in stimulating CCK secretion from EECs and that the stimulant peptides originate from dolicholin, a newly identified phaseolin-like globular protein in Country beans.     

Identification of a protein disulfide isomerase of Neospora caninum in excretory-secretory products and its IgA binding and enzymatic activities

A protein disulfide isomerase of Neospora caninum (NcPDI) with a molecular weight of 50kDa was identified in tachyzoite lysate and excretory-secretory (ES) products. The IgA antibody in 58.0% of the individual cattle tear samples recognized the NcPDI, which suggests that the PDI-specific antibody may be involved in defense against parasites. In addition, PDI-specific inhibitors and NcPDI antiserum showed inhibitory effects on the growth of N. caninum tachyzoites. Furthermore, the purified recombinant NcPDI demonstrated biological activities in vitro by catalysis and refolding of reduced RNase A and assisted in the recovery of native lysozyme. These findings indicate that NcPDI possesses PDI-specific enzymatic activity and could be a putative target for chemotherapy for neosporosis.     

The effect of 7.2% hypertonic saline solution on the duration of sodium gradient between the cerebrospinal fluid and the venous circulation in the dog

To determine the duration of water movement from cerebrospinal fluid (CSF) into venous blood by the infusion of 7.2% hypertonic saline solution (HSS), the sodium gradient between venous blood and CSF were examined. Venous sodium concentrations remained higher than that in CSF for duration of 60 min following HSS infusion. By 90 min, the CSF sodium concentration reached the equilibrium with venous sodium concentration. Those data suggests that the duration of time during which water moved from CSF into capillaries in brain by the gradient of sodium concentration was less than 90 min.     

Quantification of the polyphenols and triterpene acids in chinese hawthorn fruit by high-performance liquid chromatography

The levels of seven polyphenols (epicatechin, procyanidin B2, procyanidin B5, procyanidin C1, hyperoside, isoquercitrin, and chlorogenic acid) and two triterpene acids (oleanolic acid and ursolic acid) in the matured fruits of Chinese hawthorn (Crataegus pinnatifida Bge. var. major N.E.Br.) were determined by high-performance liquid chromatography methods. The average contents of those constituents in 37 representative cultivars were 1405, 1505, 339, 684, 56, 41, 234, 952, and 147 microg/g fresh weight (FW), respectively. A significant inverse correlation between the procyanidin contents and the latitude of the geographical origin of the cultivars was observed (r = 0.3851, P < 0.02). Correlation analysis of the levels of the nine compounds in the 37 cultivars yielded a strong correlation (P < 0.001) between the individual levels of the four procyanidins and the sum of the procyanidins level (r = 0.7413-0.9898) and between the flavonoids and the chlorogenic acid (r = 0.5383-0.9212). The changes in level of the nine compounds in the hawthorn fruit were evaluated during maturation using the Hebei Dajinxing cultivar. Sixty-one days after blossom, the polyphenol level reached the highest point and the sum of the contents was 1.36 g/100 g FW.     

Improvement in isolation and identification of food-derived peptides in human plasma based on precolumn derivatization of peptides with phenyl isothiocyanate

For the isolation and detection of food-derived peptides in blood, an approach based on the derivatization of peptides with phenyl isothiocyanate (PITC) was developed. This approach allows hydrophilic peptides to be resolved and specifically detected by reversed-phase (RP) HPLC. For the rapid capturing and clarification of peptides in human plasma, solid-phase extraction by using a mini spin column (5 mmx5 mm) packed with a strong cation exchanger was used. The clarified peptide fraction was further fractionated by size-exclusion chromatography (SEC). The peptides in the SEC fractions were derivatized with PITC, and the derivatives were resolved by RP-HPLC by using an ammonium acetate buffer or a trifluoroacetic acid system. An automatic peptide sequencer based on Edman degradation with a modified program can directly analyze the resolved derivatives. Some synthetic peptides and food-derived peptides in human plasma were successfully isolated and identified by this approach.