Release of Bound Lipids in Cereal Starches Upon Hydrolysis by Glucoamylase

The raw starch granules from corn, rice, and wheat were hydrolyzed by practically pure glucoamylase (Rhizopus niveus). The bound lipids remaining in the residual starches were investigated, of which the major components of the lipids, free fatty acids (FFA) in corn starch, FFA and phospholipids (PL) in rice starch, and PL in wheat starch were determined. In each case, the bound FFA and PL were decreased to some extent during the initial stage of hydrolysis. During the later stages, the FFA continued to gradually decrease, while the level of PL stabilized. It was interesting that some of the bound lipids were released from the granules upon glucoamylase hydrolysis, differing from the model amylose-lipid complexes. Furthermore, the structures of the residual starches were investigated. The blue value and λ_max of the starches were increased by partial hydrolysis of the starch granules using practically pure glucoamylase. Two gel-permeation chromatography analyses revealed that the relative amount of amylose fraction was increased by glucoamylase hydrolysis, and also that the increments were reduced by the defatting of bound lipids. The results suggest that the increase in amylose fraction is attributable to the existence of bound lipids in the granules.     

AlgU contributes to the virulence of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">tomato</Emphasis> DC3000 by regulating production of the phytotoxin coronatine

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen to investigate the molecular basis of plant–pathogen interactions. The function of many potential virulence factors encoded in the Pst DC3000 genome and their modes of action are not fully understood. P. syringae is known to produce the exopolysaccharide alginate. Although AlgU, a sigma factor, is known to regulate the expression of genes such as algD related to alginate biosynthesis, the molecular mechanisms of AlgU in the virulence of Pst DC3000 is still unclear. To investigate the function of AlgU and alginate in plant–bacterial pathogen interactions, we generated ΔalgU and ΔalgD mutants. After inoculation with ΔalgU but not ΔalgD, host plants of Pst DC3000 including tomato and Arabidopsis had milder disease symptoms and reduced bacterial populations. Expression profiles of Pst DC3000 genes revealed that AlgU can regulate not only the expression of genes encoding alginate biosynthesis, but also the expression of genes related to type III effectors and the phytotoxin coronatine (COR). We also demonstrated that the ΔalgU mutant showed full virulence in the Arabidopsis fls2 efr1 double mutant, which is compromised in the recognition of PAMPs. Further, the application of COR was able to restore the phenotype of the ΔalgU mutant in the stomatal response. These results suggest that AlgU has an important role in the virulence of Pst DC3000 by regulating COR production.     

Work-related increases in titer of Campylobacter jejuni antibody among workers at a chicken processing plant in Miyazaki prefecture,Japan, independent of individual ingestion of edible raw chicken meat

Workers in poultry abattoirs may be frequently exposed to Campylobacter jejuni, which is a leading cause of bacterial food poisoning in Japan. The present study was conducted to measure the titers of IgG and IgA antibodies against C. jejuni among 104 female workers in a chicken processing plant in Miyazaki prefecture, Japan. Information regarding habitual ingestion of raw chicken meat and potential occupational risk factors was collected using a questionnaire. Acid extracts of four C. jejuni strains representing the genotypes most dominant in Miyazaki were used as antigens. The levels of both immunoglobulins measured by ELISA were not correlated with ingestion of edible raw chicken meat, the amount consumed in one sitting, or its frequency. Although age was correlated with antibody levels, the length of employment was not. Furthermore, the IgG and IgA levels in workers at the evisceration step were significantly higher than those at other locations in the plant. To identify the bacterial proteins recognized by the workers’ IgG and IgA antibodies, Western blotting followed by LC/MS was conducted. Flagellin was identified as the common protein recognized in the sera of workers for whom ELISA demonstrated both the highest and lowest antibody levels. We concluded that the titers of IgG and IgA against C. jejuni in workers at the processing plant had been increased by occupational exposure to Campylobacter, regardless of raw chicken meat ingestion.     

Difference in ultraweak photon emissions between sulfonylurea-resistant and sulfonylurea-susceptible biotypes of Scirpus juncoides following the application of a sulfonylurea herbicide

We examined the ultraweak photon emissions from a paddy weed, Scirpus juncoides , to assess the availability of photon emissions for the identification of weed biotypes resistant to sulfonylurea herbicides. The emission intensity from the plant organs increased when treated with a sulfonylurea herbicide in a concentration-dependent manner. The increment in emissions was higher in the sulfonylurea-resistant biotypes than in the sulfonylurea-susceptible biotypes. The difference between the biotypes was greater in the culms than in the roots and remained so through the vegetative growth stage to the flowering stage. This difference was independent of the seed source or mutations in the acetolactate synthase genes of the resistant biotypes. These results suggest that the determination of ultraweak photon emissions can be a useful method for identifying the sulfonylurea-resistant biotypes of S. juncoides.     

Screening of weed extracts for antifungal properties against Colletotrichum lagenarium,the causal agent of anthracnose in cucumber

The antifungal activity of the leaf extracts from 203 weed species was investigated by performing a bioassay using cucumber plants and Colletotrichum orbiculare. The leaf extracts from four families, namely, Urticaceae, Onagraceae, Commelinaceae, and Solanaceae, showed a relatively stronger inhibition of the anthracnose lesions in cucumber plants when compared with the other families investigated in the study. A remarkable inhibition of anthracnose infection in cucumber leaves was observed with the extracts from the following 19 weed species: Boehmeria nipononivea and Boehmeria longispica, Persicaria scabra, Ranunculus japonicus and Ranunculus sceleratus, Cardamine flexuosa, Oenothera biennis, Aeschynomene indica, Indigofera pseudo‐tinctoria, Torilis scabra, Calystegia japonica, Solanum americanum, Bidens pilosa, Gnaphalium japonicum, Kalimeris yomena, Bromus catharticus, Cynodon dactylon, Alopecurus aequalis, and Scirpus tabernaemontani. In particular, it is noteworthy that the extracts from C. dactylon, K. yomena, and S. americanum completely inhibited anthracnose infection in cucumber.     

Development and introduction of detection dogs in surveying for scats of small Indian mongoose as invasive alien species

The small Indian mongoose was introduced to the island of Okinawa in 1910 to control rats and snakes, however, it poses a threat to many endemic species in the Yambaru forest region in Okinawa Island. An important concern in eradication is that, as much as possible, capture should be carried out without affecting native species. To protect endemic species in the Yambaru region of Okinawa Island, we trained and used detection dogs to search for mongoose scats that indicated that mongoose were present in the area. We confirmed the ability of the dogs to detect scats in positive and negative control tests in the Yambaru forest region after basic, obedience, and mongoose-scat-detection specific training. In 30 trials, the dogs found 60 of 65 scats samples. For the trained alert, the dogs carried out 59 trained alerts for the 60 scats found. In the positive and negative control tests, the dogs showed no reaction to 129 of 130 negative controls. We then surveyed the forest in the Yambaru region for mongoose scats using the detection dogs. In this study, the detection dogs found 79 scats in a survey of a 69.0-km area. We divided the survey area into 43 grids and calculated the rate of detection in each grid. The rate of detection of scats was higher in the southern part of the Yambaru region. These result showed that our mongoose scat detection dogs can be useful tools to assist in determining the presence of mongoose in the Yambaru region of Okinawa Island.     

Neutropenia associated with osteomyelitis due to Hepatozoon canis infection in a dog

A 4-year-old, intact male Shiba dog was referred to Yamaguchi University Animal Medical Center, Yamaguchi, Japan, for the following complaints: anorexia, lethargy, intermittent fever, gingival bleeding and abdominal purpura. The dog presented with persistent neutropenia. Histopathological examination of a bone marrow sample revealed round to oval structures that resembled Hepatozoon micromerozoites and formed a "wheel-spoke" pattern. Furthermore, mature neutrophils were observed around these structures. PCR and sequencing using bone marrow aspirate confirmed Hepatozoon canis (H. canis) infection. These findings suggest that the neutropenia observed in this case was associated with osteomyelitis due to H. canis infection. This is the first report of neutropenia associated with H. canis infection. H. canis infection can be included in the differential diagnosis in canine cases of neutropenia in areas where the disease is endemic.     

Two forms of cytochrome P450 cDNA from 3-methylcholanthrene-treated European eel Anguilla anguilla

ABSTRACT:     The cytochrome P450 (CYP) represents a large group of microsomal monooxygenases that catalyze drugs as well as a host of lethal environmental contaminants such as dioxins, leading to either detoxification and excretion from the animal or generation of carcinogenic intermediates. In the present study two forms of cDNA were cloned (Eu MC1 and Eu MC2) for European eel CYP1A genes by polymerase chain reaction (PCR) techniques. The cDNA of Eu MC1 was 3368 bp long coding 521 amino acid residues, and that of Eu MC2 was 2464 bp long coding 517 amino acid residues. Identities of deduced amino acid sequences between Eu MC1 and Japanese eel CYP1A1 and that between Eu MC2 and the second form of Japanese eel CYP1A were 98% and 97%, respectively, showing decisively that Eu MC1 and Eu MC2 are orthologous to Japanese eel CYP1A1 and the second form of CYP1A, respectively. A striking difference between the two eel species was that the Eu MC1 peptide was two amino acid residues longer than that of the Japanese eel CYP1A1. Existence of two loci of CYP1A in Japanese and European eels may suggest that the two forms of CYP1A exist widely among the eel species, because the divergence between the two eel species has been shown to be close to the basal divergence among eels. The identities in CYP1A may help to estimate genetic distance between European and Japanese eels.     

Cytotoxic constituents in the holdfast of cultivated Laminaria japonica

ABSTRACT:     Four steroidal ketones were isolated from the holdfast of cultivated Laminaria japonica . The structures were characterized as ergosta-4,24(28)-diene-3-one (1), ergosta-4,24(28)-diene-3,6-dione (2), stigmasta-4,24(28)-diene-3-one (3) and stigmasta-4,24(28)-diene-3,6-dione (4), by spectral data. Compounds 2 and 4 were shown to be cytotoxic against the MCF-7 human breast cancer cell, and the growth of MCF-7 was inhibited by 96% and 79%, respectively, at 10 µg/mL. It is the first report on the isolation of cytotoxic steroidal ketones from the kelp in the genus of Laminaria.