Effect of the methoxyiminoacetamide fungicide,SSF129, on respiratory activity in Botrytis cinerea

(E)-2-Methoxyimino-N-methyl-2-[2-(2,5-dimethylphenoxymethyl)phenyl]acetamide (SSF129) has been developed as a broad-spectrum systemic fungicide for control of cereal and fruit diseases. This compound inhibited NADH-oxidation by submitochondrial particles from mycelial cells of Botrytis cinerea, with an EC₅₀ value of 14.5 nM , due to blockage of electron transport through the cytochrome bc₁ complex in the mitochondrial respiratory chain. However, SSF129 did not suppress, but rather increased, oxygen consumption by mycelial cells of the fungus. This was because mycelial cells contain an alternative oxidase protein and the cells have the ability to rapidly switch electron flux from the main cytochrome pathway to the alternative pathway on blockage of the former by SSF129. The alternative pathway of the mycelia seems not to be operative when the cytochrome pathway is functional. Naturally occurring flavonoids inhibited the alternative oxidase of the mycelial cells in a dose-dependent manner, with EC₅₀ values of 68.4 µM for flavone and 63.7 µM for flavanone. These observations suggested that plant components play an important role in control of gray mould by SSF129. © 1999 Society of Chemical Industry     

Measurements of density contrast and sound-speed contrast for target strength estimation of Neocalanus copepods (Neocalanus cristatus and Neocalanus plumchrus) in the North Pacific Ocean

The mass density and sound-speed contrasts against surrounding seawater (g and h, respectively) of Neocalanus copepods (N. cristatus and N. plumchrus) were measured in 2006 and 2007 to compute the theoretical target strength (TS). The values of g ranged from 0.997 to 1.009 in N. cristatus and from 0.995 to 1.009 in N. plumchrus. There were no correlations between prosome length (PL) and g. The values of h ranged from 1.006 to 1.021 in N. cristatus and from 1.013 to 1.025 in N. plumchrus and varied with changes in temperature. TS was estimated with the theoretical sound scattering model using the values of g and h based on the temperature, salinity, and depth of the location where the specimens were collected. Regressions of the tilt-averaged TS versus PL were obtained at 38, 120, and 200 kHz. The averaged TS of N. cristatus and N. plumchrus at 120 kHz, which is widely used as a high frequency, ranged from −110.0 to −103.1 dB and from −121.4 to −109.7 dB, respectively. There was a positive correlation between frequency and averaged TS: the higher the frequency, the higher the value of averaged TS. The TS at 120 and 38 kHz varied from 14.8 to 16.4 dB in N. cristatus and from 17.9 to 18.7 dB in N. plumchrus, respectively; that at 200 and 120 kHz varied from 2.9 to 5.5 dB in N. cristatus and from 5.3 to 6.5 dB in N. plumchrus, respectively.     

cDNA cloning and primary structure comparison of tauropine dehydrogenase and β-alanopine dehydrogenase from the limpet <Emphasis Type="Italic">Cellana grata</Emphasis>

The primary structures of β-alanopine dehydrogenase (β-AlDH) and tauropine dehydrogenase (TaDH) from the limpet Cellana grata were determined by amino acid sequence analysis and complementary DNA (cDNA) cloning. β-AlDH and TaDH cDNAs comprised 1,479 nucleotides and 1,444 nucleotides, respectively, and both included an open reading frame of 1,206 nucleotides corresponding to 402 amino acids. The enzymes showed very high homology, with 96% amino acid identity. These enzymes were homologous to other marine invertebrate opine dehydrogenases (OpDHs), except TaDH of the marine sponge Halichondria japonica. The highest homologies were to alanopine dehydrogenase from Fusitriton oregonensis, being 57% for both enzymes. A phylogenetic tree constructed on the basis of marine invertebrate OpDHs and developed using a sequence distance method and neighbor-joining algorithm showed a tendency for the classification of animals from taxonomically derived evolutionary trees. Additionally, Cellana grata OpDHs belong to the same group as the Gastropoda OpDHs. This represents the first report concerning the primary structure of marine invertebrate β-AlDH, and primary structure comparisons of clearly different enzymes from the same species.     

Endocarditis in chickens caused by subclinical infection of Streptococcus gallolyticus subsp. gallolyticus

Streptococcus gallolyticus subsp. gallolyticus causes endocarditis in humans and acute septicemia in domestic birds. We describe here the infective endocarditis caused by the bacterium found among clinically healthy broilers at two abattoirs in Japan. The chickens were thought to be healthy because of the lack of clinical symptoms and normal levels of mortality before slaughtering. At the time of inspection, some chickens were condemned because of organ disorders characterized by vegetative valvular endocarditis as well as focal necrosis in heart, liver, and spleen. Streptococcus gallolyticus subsp. gallolyticus was isolated from the organs as a pure culture, indicating that the bacterium probably was the causative agent of the disorders. Amplified fragment length polymorphism analysis of the isolates collected at the abattoirs from chickens grown in nine different farms indicated that the isolates were different variants of the same clonal lineage and may have been derived from the same ancestor. These results suggest that S. gallolyticus subsp. gallolyticus causes infectious endocarditis in chickens and that healthy chickens may possess the bacterium in their normal flora as an opportunistic pathogen.     

Indole-3-carbaldehyde: a tyrosinase inhibitor from fungus YL185

Indole-3-carbaldehyde (1) was isolated as a tyrosinase inhibitor from the ethyl acetate-soluble fraction of extracellular fluids of unknown fungus YL185. The partial sequencing data of 18S ribosomal DNA (18S rDNA) indicate that this isolate belongs to the family Polyporaceae or Corticiaceae sensu lato. Indole-3-carbaldehyde inhibited the oxidation of l-3,4-dihydroxyphenylalanine (l-DOPA) by mushroom tyrosinase with a 50% inhibitory concentration (IC₅₀) of 1.3mM and showed inhibitory activity on melanin production in B16 melanoma cells. The aldehyde group of 1 plays an important role in eliciting tyrosinase inhibitory activity.     

Evidence for existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa

Cyclic adenosine 3',5'-monophosphate (cAMP) signaling regulates the expression of fertilizing ability in mammalian spermatozoa. Many articles indicate that this signaling is mediated mainly via protein kinase A. Recently, a guanine nucleotide exchange factor for small G protein Rap1 (an exchange protein directly activated by cAMP: Epac) was discovered as a new mediator of cAMP signaling in somatic cells. The aim of this study was to reveal the existence of cAMP-Epac signaling in mouse spermatozoa. Northern blot analysis and in situ hybridization suggested that Epac1 and Epac2 mRNAs were transcribed in the seminiferous epithelia of the testis. This shows that expression of Epac mRNAs is present in mouse testicular germ cells. Indirect immunofluorescence with specific polyclonal antibodies suggested possible co-localization of Epac1 and Rap1 proteins in the heads of epididymal spermatozoa. Moreover, treatment of epididymal spermatozoa with an Epac-specific cAMP analog, 8-pMeOPT-2'-O-Me-cAMP, induced activation of Rap1, as revealed with a commercial kit for pull-down assay. These results indicate the existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa.     

A 32-kDa tyrosine-phosphorylated protein shows a protease-dependent increase in dead boar spermatozoa

Boar sperm TyrP32 is a 32-kDa tyrosine-phosphorylated protein that increases during the capacitation and acrosome reaction and during cryocapacitation. However, it is still unclear whether the increase in TyrP32 is an event that is limited to the process of sperm fertilization, including cryocapacitation. The aims of the present study were to demonstrate that TyrP32 is increased in dead spermatozoa after freeze-thawing without a cryoprotectant and to find the causal factors for this increase. Washed spermatozoa were resuspended in a salt solution and then frozen. The frozen samples were rapidly thawed in a warm water bath and then used for sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE)/Western blotting to detect TyrP32, SDS-PAGE/silver staining of sperm proteins and staining of acrosomal contents with fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA). In the samples before freezing, TyrP32 was barely detectable, and the distribution of the acrosomal contents was normal in most spermatozoa. One cycle of freeze-thawing induced an increase in TyrP32, a decrease in major sperm proteins and disorder in the acrosomal contents. However, the addition of a protease inhibitor (APMSF, 1 mM) suppressed the increase in TyrP32 and the decrease in the major sperm proteins, although it did not have any influence on the disorder in the acrosomal contents. Additionally, the spermatozoa did not exhibit any flagellar movement after freeze-thawing, which showed that almost all of them were dead. These results indicate that TyrP32 can show a protease-dependent increase in dead spermatozoa after freeze-thawing without a cryoprotectant even though the dead spermatozoa do not undergo cryocapacitation.     

Modulation of gut microbiota associated with abalone Haliotis gigantea by dietary administration of host-derived Pediococcus sp. Ab1

In the present study, we evaluated how dietary administration of host-derived Pediococcus sp. Ab1 has an effect on the abalone gut microbiota using a culture-dependent method and 16S rRNA gene library analysis. The culturable lactic acid bacteria number in the probiotic sample was 10⁵ higher than that in the non-probiotic sample, and we speculate that this significant increase was due to colonization of Ab1 into abalone gut. The result of a culture-dependent method showed that the proportion of Vibrio halioticoli clade, which is known to be a beneficial resident bacterium to abalone, was much higher in the probiotic sample than in the non-probiotic sample. 16S rRNA gene clone sequences revealed that gut microbiota in the probiotic sample was obviously diverse compared to the non-probiotic sample, probably due to improvement of the gut environment by Ab1 colonization. In addition, some beneficial bacteria–like sequences such as V. halioticoli were only found in the probiotic sample. These results suggest that the dietary administration of Ab1 to abalone gut has a great effect on modulation of not only culturable but also unculturable gut microbiota. Our results are useful for future investigations into understanding the effect of probiotics on gut microbiota.