Mycobacterium pseudoshottsii isolated from 24 farmed fishes in western Japan

Mycobacteria isolated from epizootics of farmed fishes in western Japan were examined for the first time using multigenotypic analysis. By analysis of the sequences of the internal transcribed spacer between the 16S and 23S rRNA genes (ITS) region and the partial 16S rRNA, hsp65 and rpoB genes, M. pseudoshottsii was identified as the causative agent in these infections. Prior to this study, only M. marinum has been known as the causative agent of lethal mycobacterial disease in marine fishes in Japan.     

Neutrophils contact to plasma membrane of keratinocytes including desmosomal structures in canine pemphigus foliaceus

Pemphigus foliaceus (PF) is an autoimmune blistering skin disease that affects certain mammals including dogs. In canine PF, neutrophils are infiltrated intensely into pustular lesions including acantholytic cells, although neutrophilic infiltration is not characterized in human PF. The roles of the neutrophils in the cutaneous lesions of canine PF have not yet been understood. The purpose of this study was to characterize the ultrastructural features underlying the acantholysis with pustule formation in canine PF. Four dogs diagnosed as PF on the basis of clinical signs, histopathological findings, and direct and indirect immunofluorescence examinations were performed. Electron microscopy revealed that the acantholytic cells were adjacent to multiple neutrophils in the pustules. At the contact points between neutrophils and acantholytic keratinocytes, half-desmosomes of acantholytic keratinocytes with intact attachment plaques were observed within invaginations of neutrophils. Furthermore, on the surface of acantholytic cells in the pustules, neutrophil granules seemed to be secreted to the surface of acantholytic cells and to degenerate the half-desmosome structures. Neutrophils were also observed within the epidermis adjacent to the pustule. At the intercellular gap between two dissociated keratinocytes, neutrophils inserted its pseudopodia into the gap between the two half-desmosomes of keratinocytes. These findings taken together suggested that, at least in the areas where we analyzed ultrastructurally, neutrophils contact desmosomal structures and seem to play some parts in separation of keratinocytes and degeneration of split-desmosomes in pustules of dogs with PF.     

Calmodulin is involved in the occurrence of extracellular Ca2+-dependent full-type hyperactivation in boar ejaculated spermatozoa incubated with cyclic AMP analogs

In mammals, hyperactivation is essential for sperm fertilization with oocytes in vivo. Two types of hyperactivation “full-type and nonfull-type patterns” can be observed in the spermatozoa from boars, bulls, and mice. We have a hypothesis that the full-type hyperactivation is a physiological (in vivo) pattern and are elucidating its molecular bases. The aims of this study were to detect calmodulin in boar sperm flagella by Western blotting and indirect immunofluorescence and to investigate effects of extracellular Ca²⁺ and calmodulin antagonists “W-7 and W-5 (W-5; a less potent antagonist)” on the occurrence of full-type hyperactivation in boar spermatozoa. Calmodulin was specifically detected as the 17-kDa antigen in the flagella and postacrosomal region of the heads. Full-type hyperactivation could be induced effectively in the samples incubated with 3.42 mM CaCl₂ for 120–180 min, and it was significantly reduced in the concentration-dependent manners of W-7 and W-5. Suppressing effects of W-7 on the full-type hyperactivation were stronger than those of W-5. These observations indicate that flagellar calmodulin is involved in the occurrence of extracellular Ca²⁺-dependent full-type hyperactivation in boar spermatozoa. This is the first indication of the intracellular Ca²⁺-sensing molecule which can function in the full-type hyperactivation.     

Effects of Acid Stress on Reproductive Functions in Immature Carp, Cyprinus Carpio

Changes in plasma cortisol, estoradiol-17β (E2) and 11-ketotestosterone (11-KT) levels under chronic acid exposure were examined to clarify the endocrinological effects of lowered environmental pH on gametogenesis in immature and gonadal developing carp, Cyprinus carpio. Plasma cortisol levels increased markedly 3 hours after the acid exposure and high levels were maintained for 48 hours, and then gradually decreased. Plasma E2 levels were significantly elevated and peaked at 1 week after the acid exposure. After that, E2 levels decreased gradually, but remained at significantly higher levels than the control group even at 4 weeks after the exposure. Plasma 11-KT levels showed no significant change throughout the experiment. These results indicate that acidic water disrupts the endocrine system that may have adverse effects on gametogenesis in freshwater fish.     

Behaviors of D- and L-lactic acids during the brewing process of sake (Japanese rice wine)

The amounts of D- and L-lactic acids during the brewing process of sake were determined by capillary electrophoresis using 2-hydroxypropyl-beta-cyclodextrin as a chiral selector. Because L-lactic acid, which prevents the growth of nonuseful microorganisms, is a raw material of sake, the ratio of L-lactic acid to total lactic acid is almost 1.0 at the initial stage of sake brewing. During brewing, the ratio decreased gradually and finally reached 0.39. Yeast (Saccharomyces cerevisiae) for sake brewing produced D-lactic acid, but not L-lactic acid in a culture medium. These results suggest that the decrease in the ratio of L-lactic acid to total lactic acid during sake brewing resulted in D-lactic acid production by yeast. The ratios in 18 brands of sake obtained commercially ranged from 0.23 to 0.78. The levels of D-lactic acid in sake (140-274 mg/L) were in a narrower range than those of L-lactic acid (61-461 mg/L). Although the D-lactic acid level in sake did not correspond to total lactic acid level, the L-lactic acid level correlated well with total lactic acid level (R(2) = 0.867). These results suggest that the ratio of L-lactic acid to total lactic acid in sake reflected the amount of L-lactic acid added at the initial stage of sake brewing.     

A preliminary investigation on microstructural characteristics of interfacial zone between cement and exploded wood fiber strand by using SEM-EDS

The hydration behavior and strength performance of cement mixed with exploded wood fiber strand (WFS) obtained by the water-vapor explosion process have been studied previously. In the current study, the microstructural characteristics of cement–exploded WFS interfacial zone were examined using scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDS). The Ca/Si ratios at the interfacial zones and the elemental compositions of hydration products deposited in the tracheid lumen were investigated. In addition, the morphological differences and compositional variations of hydration products that developed on the wood surfaces were examined. The results revealed that the Ca/Si ratios at the interfacial zones were strongly influenced by the mixture compositions, and that the elemental compositions of the hydration products that filled the tracheid lumen were significantly different from those of the cement paste in the mixtures. Differences in morphology and composition of hydration products at the wood surfaces were also observed to correspond to the different mixture compositions. These characteristics are considered to be directly related to the bond property, and thus, to the mechanical performance of WCM.Part of this report was presented at the 53rd Annual Meeting of the Japan Wood Research Society, Fukuoka, March 2003     

Suppressive mechanism of seed coat pigmentation in yellow soybean

In soybean seeds, numerous variations in colors and pigmentation patterns exist, most of which are observed in the seed coat. Patterns of seed coat pigmentation are determined by four alleles (I, iⁱ, i^k and i) of the classically defined I locus, which controls the spatial distribution of anthocyanins and proanthocyanidins in the seed coat. Most commercial soybean cultivars produce yellow seeds with yellow cotyledons and nonpigmented seed coats, which are important traits of high-quality seeds. Plants carrying the I or iⁱ allele show complete inhibition of pigmentation in the seed coat or pigmentation only in the hilum, respectively, resulting in a yellow seed phenotype. Classical genetic analyses of the I locus were performed in the 1920s and 1930s but, until recently, the molecular mechanism by which the I locus regulated seed coat pigmentation remained unclear. In this review, we provide an overview of the molecular suppressive mechanism of seed coat pigmentation in yellow soybean, with the main focus on the effect of the I allele. In addition, we discuss seed coat pigmentation phenomena in yellow soybean and their relationship to inhibition of I allele action.     

Relationship between spike morphology and habitat of four <Emphasis Type="Italic">Aegilops</Emphasis> species of section Sitopsis

This study examines the relationship between spike morphology and natural habitat for 84 accessions of four Aegilops species, belongs to section Sitopsis, Ae. bicornis, Ae. longissima, Ae. searsii, and Ae. sharonensis in genus Aegilops, section Sitopsis, wild relatives of Triticum aestivum L. These species are considered valuable genetic resources for future cultivation and breeding of domesticated wheat. The goals of the study were to: (1) document variation in spike morphology among these four species; (2) examine the relationship between spike morphology and native habitat; (3) document geographical distribution of distinct spike morphology; and (4) examine the relationship between spike morphology and heading time and value for these four species. The results reveal significant differences in spike morphology among species of section Sitopsis. The most noteworthy variation involved the absence/presence of lateral awn, such that species with lateral awn were restricted in coastal, though species without lateral awn were mainly distributed in inland. This suggests that local climate may be a determinant of variation in lateral awn, and that this trait may be subject to convergent evolution. Differences in heading time in sympatric area were also observed. The differences may enhance species divergence and could represent a lead speciation event. The results of this study will facilitate identification of populations or accessions of wild wheat with favorable traits and/or novel adaptive genes.     

Protective effects of dietary chamomile tea on diabetic complications

Matricaria chamomilla L., known as "chamomile", has been used as an herbal tea or supplementary food all over the world. We investigated the effects of chamomile hot water extract and its major components on the prevention of hyperglycemia and the protection or improvement of diabetic complications in diabetes mellitus. Hot water extract, esculetin (3) and quercetin (7) have been found to show moderate inhibition of sucrase with IC50 values of 0.9 mg/mL and 72 and 71 microM, respectively. In a sucrose-loading test, the administration of esculetin (50 mg/kg body weight) fully suppressed hyperglycemia after 15 and 30 min, but the extract (500 mg/kg body weight) and quercetin (50 mg/kg body weight) were less effective. On the other hand, a long-term feed test (21 days) using a streptozotocin-induced rat diabetes model revealed that the same doses of extract and quercetin showed significant suppression of blood glucose levels. It was also found that these samples increased the liver glycogen levels. Moreover, chamomile extract showed potent inhibition against aldose reductase (ALR2), with an IC50 value of 16.9 microg/mL, and its components, umbelliferone (1), esculetin (3), luteolin (6), and quercetin (7), could significantly inhibit the accumulation of sorbitol in human erythrocytes. These results clearly suggested that daily consumption of chamomile tea with meals could contribute to the prevention of the progress of hyperglycemia and diabetic complications.     

A comparison of the test tube and the dialysis tubing in vitro methods for estimating the bioavailability of phosphorus in feed ingredients for swine

The validity of a simplified in vitro test tube (TT) method was compared with a more complicated dialysis tubing (DT) method to estimate the percentage of available phosphorus (P) in 41 plant origin and five animal origin feed ingredients for swine. The TT method using 1.0 or 0.25 g samples was compared with the DT method using 1.0 g samples at two pancreatic incubation times (2 vs 4 h) in a 3 x 2 factorial arrangement of treatments. Each DT and TT method treatment was replicated three and six times, respectively. Both methods utilize three enzymatic digestions: (i) predigestion with endoxylanase and beta-glucanase for 1 h, (ii) pepsin digestion for 2 h, and (iii) pancreatin digestion for 2 or 4 h. For the TT method, the entire procedure was conducted in a 50 mL conical centrifuge tube and replicated six times. For the DT method, the first two digestions were conducted in a 10 mL plastic syringe before the contents were quantitatively transferred into a segment of DT for the pancreatic digestion. The percentages of hydrolyzed P for plant origin ingredients measured by the DT method using 1.0 g samples and the TT method using 0.25 g samples were highly correlated (r = 0.94-0.97, P < 0.001) with each other and with published in vivo available P values for swine. Repeatabilities for these two methods ranged from 99.64 to 99.86%. The TT method using 1.0 g samples, however, did not provide valid estimates of P availability for all ingredients. For animal origin ingredients, neither method was significantly correlated (r = 0.1-0.6, P >or = 0.4) with published in vivo available P values. In conclusion, the accuracy and validity of the TT method using 0.25 g samples with a 2 h pancreatic digestion time was equal to or superior to the DT method using 1.0 g samples with a 4 h pancreatic digestion time for estimating P availability in plant origin feed ingredients.