Automatic counting of FISH-labeled microbes by an LED illuminated detecting apparatus

ABSTRACT:   The Bioplorer (BP) (Matsushita Ecology Systems Co. Ltd, Kasugai, Aichi-ken, Japan) is an apparatus that consists of a light-emitting diode and optical and image analysis systems. This instrument has been developed and used for the semi-automatic counting of total microbes, mainly in food, cosmetics and industrial products. In this study, the applicability of BP to the detection and enumeration of eukaryotic and prokaryotic cells, after the fluorescent in situ hybridization (FISH) procedure, was examined. The cells of a yeast ( Candida albicans ) and a bacterium ( Escherichia coli ) were specifically labeled using an oligonucleotide probe with fluorochrome and then counterstained with 4', 6-diamidino-2-phenylindole (DAPI). The numbers of cells after FISH treatment and DAPI staining agreed well, indicating that the BP optical device and image analysis system had enough sensitivity to detect and quantify eukaryotic and prokaryotic cells. Because of its simplicity and reliability, BP can be a new tool for quantification of specifically labeled target microorganisms, both in industrial products and natural samples.     

Effect of blue and red LED light irradiation on β-cryptoxanthin accumulation in the flavedo of citrus fruits

β-Cryptoxanthin (β-cry), an antioxidant abundant in citrus fruits, plays an important role in the prevention and treatment of certain diseases, especially cancers. In the present study, to increase the content of β-cry in citrus flavedo, the effects of blue (470 nm) and red (660 nm) light-emitting diode (LED) lights on the accumulation of carotenoids and expression of genes related to carotenoid biosynthesis were investigated in the flavedo of Satsuma mandarin. The results showed that accumulation of β-cry was induced by red light, while it was not affected by blue light. The accumulation of β-cry under red light was attributed to simultaneous increases in the expression of CitPSY, CitPDS, CitZDS, CitLCYb1, CitLCYb2, CitHYb, and CitZEP. The results presented herein might provide new strategies to enhance the commercial and nutritional value of citrus fruits.     

Responses of fees used for revegetation of desert areas to iron deficiency stress

We investigated the responses to iron deficiency of four major tree species, Prosopis cineraria (local name: Ghaf), Acacia tortilis (Samar), Zizyphus spina-christi (Sidr), and Leptadenia pyrotechnica (Markh), used for revegetation of desert areas in the United Arab Emirates (UAE). The responses to iron deficiency differed among the tree species. Ghaf, Samar, and Sidr decreased the nutrient solution pH in response to iron deficiency. Markh and iron-deficient Sidr did not release protons in the CaCl₂ solution. The Fe reducing capacity of the roots of Ghaf, Samar, and Sidr was significantly enhanced by iron deficiency, whereas a smaller increase in the reducing activity of the roots of Markh was observed. In all the tree species, the amount of reductant released from iron-deficient plants was higher than that from iron-sufficient ones. Markh released a small amount of reductant under irondeficient conditions. In the present study, the reductant released by all the trees was caffeic acid. Acidification treatment enhanced the amount of reductant released irrespective of iron treatments in Ghaf, Samar, and Sidr but had no effect on the amount of reductant released in iron-deficient Markh. The activity of p-coumarate hydroxylase was higher in Sidr and Ghaf than in the other trees. This activity decreased in iron-deficient Samar. Ghaf, Samar, and Sidr showed a high capacity to lower the pH of the nutrient solution, and Ghaf markedly enhanced the effectiveness of all the examined mechanisms, while Markh showed a lower ability compared to the other tree species.     

Comparison of the effects explained by variations in the bovine PLAG1 and NCAPG genes on daily body weight gain,linear skeletal measurements and carcass traits in Japanese Black steers from a progeny testing program

The c.1326T>G single nucleotide polymorphism (SNP) in the NCAPG gene, which leads to an amino acid change of Ile442 to Met442, was previously identified as a candidate causative variation for a bovine carcass weight quantitative trait loci (QTL) on chromosome 6, which was associated with linear skeletal measurement gains and daily body weight gain at puberty. Recently, we identified the stature quantitative trait nucleotides (QTNs) in the PLAG1‐CHCHD7 intergenic region as the causative variations for another carcass weight QTL on chromosome 14. This study aimed to compare the effects of the two QTL on growth and carcass traits using 768 Japanese Black steers from a progeny testing program and to determine whether a genetic interaction was present between them. The FJX_250879 SNP representing the stature QTL was associated with linear skeletal measurements and average daily body weight gain at early and late periods during adolescence. A genetic interaction between FJX_250879 and NCAPG c.1326T>G was detected only for body and rump lengths. Both were associated with increased carcass weight and Longissimus muscle area, and NCAPG c.1326T>G was also associated with reduced subcutaneous fat thickness and increased carcass yield estimate. These results will provide useful information to improve carcass weight in Japanese Black cattle.     

Estimation of variance components for carcass traits in Japanese Black cattle using 50K SNP genotype data

Genomic selection using high‐density single nucleotide polymorphism (SNP) genotype data may accelerate genetic improvements in livestock animals. In this study, we attempted to estimate the variance components of six carcass traits in fattened Japanese Black steers using SNP genotype data. Six hundred and seventy‐three steers were genotyped using an Illumina Bovine SNP50 BeadChip and phenotyped for cold carcass weight, ribeye area, rib thickness, subcutaneous fat thickness, estimated yield percent and marbling score. Additive polygenic variance and the variance attributable to a set of SNPs that had statistically significant effects on the trait were estimated via Gibbs sampling with two models: (i) a model with the chosen SNPs and the additive polygenic effects; and (ii) a model with the polygenic effects alone. The proportion of the estimated variance attributable to the SNPs became higher as the number of SNP effects that fit increased. High correlations between breeding values estimated with the model containing the polygenic effect alone and those estimated by chosen SNPs were obtained. No fraction of the total genetic variance was explained by SNPs associated with the trait at P ≥ 0.1. Our results suggest that for the carcass traits of Japanese Black cattle, a maximum of half of the total additive genetic variance may be explained by SNPs between 100 several tens to several 100s.     

Effects of helical angle of router bit on acoustic emission

The purpose of this study was to investigate the effects of changes in the helical angle of the router bit on the acoustic emission (AE) signal for various workpiece grain angles. The helical angle varied from 0° to 50° at 10° increments. The workpiece grains were oriented from 0 degrees (cutting parallel to the grain), through 90° (cutting end of the grain), to 165° at 15° increments. The AE signals and machined surface roughness were measured in an attempt to clarify the relations between them. The results were summarized as follows: (1) The AE signals were lowest in the with the grain cutting zones and slightly increased in the against the grain cutting zones; they rapidly reached the highest values at the 135°–165° grain angles. The greater the helical angle of the router bit, the smaller were the AE count rates for each grain angle investigated. There was no significant change in AE generation for helical angles of 0° and 10°. Moreover, the greater the feed rate, the greater was the AE count rate for every cutting condition investigated. (2) The surface roughness, similar to the AE count rate, had the lowest values in the with the grain cutting zones, slightly increased until the 120° grain angle, and then rapidly become extremely rough, reaching a maximum at the grain angles of 135°–150°. There was no remarkable change in the machined surface roughness while routing with the grain using the router bit of greater helical angle. However, when routing against the grain, the greater the helical angle the smoother was the machined surface. (3) There were correlations between the AE count rate and the machined surface roughness for each helical angle investigated. Therefore, acoustic emission has shown promise for monitoring and controlling the routing operation, including various grain angles and helical angles of the router bit.     

Sterol ferulates, sterols, and 5-alk(en)ylresorcinols from wheat, rye, and corn bran oils and their inhibitory effects on Epstein-Barr virus activation

Sterol ferulate, free sterol, and 5-alk(en)ylresorcinol constituents of wheat, rye, and corn bran oils were studied. Among the sterol ferulates, one novel compound, 24-methylenecholestanol ferulate (7), along with six known compounds, namely, 24-methylcholestanol ferulate (1), 24-methylcholesterol ferulate (2), 2-methyllathosterol ferulate (3), stigmastanol ferulate (4), sitosterol ferulate (5), and schottenol ferulate (6), were isolated and characterized. Five known free sterols, namely, 24-methylcholesterol (8), stigmastanol (9), sitosterol (10), schottenol (11), and stigmasterol (12), were isolated and identified. 5-Alk(en)ylresorcinols were found in wheat and rye bran oils but not in corn bran oil. Of these, one new compound, 5-n-(2'-oxo-14'-Z-heneicosenyl) resorcinol (19), and seven known compounds, namely, 5-n-heptadecyl- (13), 5-n-nonadecyl- (14), 5-n-heneicosyl- (15), 5-n-tricosyl- (16), 5-n-pentacosyl- (17), 5-n-(14'-Z-nonadecenyl)- (18), and 5-n-(2'-oxoheneicosyl)resorcinols (20), were isolated and characterized. These compounds were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells, which is known to be a primary screening test for antitumor promoters. Four compounds, 1, 2, 4, and 11, showed potent inhibitory effects on EBV-EA induction.     

Azaphilones, furanoisophthalides, and amino acids from the extracts of Monascus pilosus-fermented rice (red-mold rice) and their chemopreventive effects

Six azaphilones, monascin (1), ankaflavin (2), rubropunctatin (3), monascorburin (4), rubropunctamine (5), and monascorburamine (6), two furanoisophthalides, xanthomonasin A (7) and xanthomonasin B (8), and two amino acids, (+)-monascumic acid (9) and (-)-monascumic acid (10), isolated from the extracts of Monascus pilosus-fermented rice (red-mold rice) were evaluated for their inhibitory effects on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in mice, on the induction of Epstein-Barr virus early antigen (EBV-EA) by TPA in Raji cells, and on the activation of (+/-)-(E)-methyl-2[(E)-hydroxy-imino]-5-nitro-6-methoxy-3-hexemide (NOR 1), a nitric oxide (NO) donor. Among the compounds tested, seven compounds (1-6 and 10) on TPA-induced inflammation, and six compounds (1, 3-5, 9, and 10) on EBV-EA activation, exhibited potent inhibitory effects. All of the compounds tested showed moderate inhibitory effects on NOR 1 activation.     

Isolation of antifouling substances from the Palauan sponge Protophlitaspongia aga

ABSTRACT: Fouling organisms cause serious problems by settling on ships' hulls, and other marine infrastructures. Organotin compounds have been developed as effective antifoulants. However, environmental problems are associated with these compounds. It is necessary to find antifouling substances that exhibit a lower toxicity than those substances currently used. A new pyrimidine derivative, 3,4,5,6-tetrahydro-6-hydroxymethyl-3,6-dimethyl-4-pyrimidinecarboxylic acid, (compound 1) and zooanemonin (compound 2) were isolated from the marine sponge Protophlitaspongia aga as antifouling substances against the barnacle Balanus amphitrite . We also found that α-nicotinamide ribose (compound 3) inhibited germination and the attachment of Ulva spores.