Transmission of mitochondrial plasmids in protoplast cell fusion between compatible monokaryons of <Emphasis Type="Italic">Lentinula edodes</Emphasis>

Electrophoretic analysis of the transmission pattern of mitochondrial plasmids in protoplast cell fusion between compatible monokaryons of Lentinula edodes indicates that three of the four plasmids carried in parental monokaryons are effectively transferred and replicated in the protoplast fusants. The two monokaryons, 1158a and 1569a, carried different plasmids that could be distinguished by a single restriction digest. Electrophoresis of intact plasmids and restriction analyses indicate that all but one of the fusants carry three of the four possible plasmids, indicating that transmission of plasmids in protoplast fusions is principally biparental in L. edodes. Thus, heterocytoplasmic cells of L. edodes can be effectively constructed by protoplast cell fusion. In addition, plasmids of the same homology group cannot coexist in the heteroplasmic cells after protoplast cell fusion. Contribution No. 382 of the Tottori Mycological Institute     

Analysis of the equine ovarian structure during the first twelve months of life by three-dimensional internal structure microscopy

A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of follicles with a diameter of ≥10 mm and the diameter of the largest follicle increased gradually after 6 months of age. On a volume basis, the medulla accounted for nearly 90% of the whole ovary at 0 to 1 months of age, but significantly decreased from 2 to 3 months of age. The volume of the cortex increased progressively after birth and reached approximately 60% of the total volume at 8 to 12 months of age. This significant development of the cortex coincided with the increased number and size of large follicles observed from 6 months of age. These results suggest that the development of the cortex plays a role in the maturation of the follicles and the equine ovary undergoes substantial morphological changes postnatally until puberty.     

Springiness of Pancake and Its Relation to Binding of Prime Starch to Tailings in Stored Wheat Flour

Wheat flours were stored at room temperature (15–25°C), 40, 60, 80, and 100°C for various times. The baking performance of these flours was then evaluated in terms of the springiness of pancakes (recovery from crushing). Baking performance improved with increased storage time at each temperature. Brabender Amylograph tests of the flours indicated that the onset temperature in viscosity decreased with increased storage time at each temperature. When the flours were fractionated by acetic acid (pH 3.5) with mortar and pestle, recoveries of the water-solubles and gluten fractions were unchanged, but recoveries of prime starch and tailings fractions changed remarkably with increased storage time. On the other hand, those changes were not observed when flours were fractionated with a Waring blender. The binding of prime starch to tailings was correlated significantly with baking performance.     

A review of tower flux observation sites in Asia

Aggregating and sharing the metadata of flux observation sites results in a strong collaboration among various fields of study. Such data sharing will also be a part of the future design of a tower flux observation network in Asia. The aim of this review is to comprehend the state of tower flux observation sites in Asia. There are 109 tower flux observation sites in Asia including 51 forest sites. There are more new sites under construction in Asia than in America and Europe. These sites range from the taiga in Siberia to the rainforest in Southeast Asia, and from the equatorial to polar Koeppen climate zones. There are many highly humid areas in Asia, not only at low latitudes but also at middle latitudes. This climate condition has developed unique vegetation such as lucidophyllous (evergreen broadleaf) forest, which is distributed in warm areas with high precipitation in the growing season. However, there are only a few observations taking place in lucidophyllous forest. Rice paddy fields are also unique land cover in Asia. It is important to accumulate long-term data for rice fields with their management records, because plant activity depends highly on both climate conditions and land-use management. Flux data, especially net ecosystem exchange and related elements, are used for widespread studies not only within the flux-research community but also in other fields of study, for example remote sensing. At present, however, both the quantity and quality of the data are not sufficient for these studies. Regarding the quantity, there are many recently established sites that have not published data yet; regarding quality, flux data include uncertainties caused by methodological problems. Flux researchers are required not only to obtain flux data but also to improve their quality. Meanwhile, data users must understand there are still uncertainties in flux data. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Tissue distribution of tetrodotoxin and its analogs in Lagocephalus pufferfish collected in Vietnam

Fisheries Science - Pufferfish belonging to Lagocephalus are composed of several species, some of which have been recognized to be non-toxic. Although fish belonging to this genus inhabit a wide...     

Molecular Phylogenetic Analysis of Ribosomal DNA Internal Transcribed Spacer Regions and Comparison of Fertility in <Emphasis Type="Italic">Phomopsis </Emphasis>Isolates from Fruit Trees

Sequences of the ribosomal DNA (rDNA) internal transcribed spacer (ITS) regions were examined to infer a molecular phylogeny of small-spored Phomopsis isolates, designated W-type (mainly white colony, weakly virulent, bearing both alpha and beta conidia at 25°C on PDA) and G-type (mainly gray colony, highly virulent, bearing only alpha conidia at 25°C on PDA), and P. amygdali from fruit trees. Phomopsis G-type and P. amygdali were a monophyletic group distinct from the W-type. The W-type isolates were divided into two monophyletic groups. Diaporthe citri, D. tanakae, P. asparagi, P. viticola, P. vitimegaspora and D. nomurai, which are morphologically distinguishable from W- and G-types, differed from the W- and G-types in molecular phylogenetic analyses. PCR-RFLP analysis of rDNA ITS regions was useful to distinguish each of the Phomopsis species and groups using three restriction enzymes. In mating tests, W-type isolates from fruit trees were heterothallic and inter-fertile even between isolates belonging to the different monophyletic groups. Isolates of the G-type and P. amygdali collected in Japan were cross-fertile. Some isolates from Lunaria annua, Ulmus glabra and Juglans regia belonged to one of the two monophyletic groups of the W-type and were cross-fertile with W-type isolates from Rosaceous fruit trees. Received 27 September 1999/ Accepted in revised form 27 January 2000     

Reduction in the IgE reactivity of Pacific mackerel parvalbumin by mutations at Ca2+-binding sites

ABSTRACT:   Parvalbumin is a sarcoplasmic Ca²⁺-binding protein of 12 kDa and represents the major fish allergen. Several peptide segments are identified as immunoglobulin E (IgE)-binding epitopes of cod parvalbumin. However, carp parvalbumin (Cyp c 1) shows a markedly reduced IgE-binding ability upon depletion of Ca²⁺, suggesting the importance of conformational epitopes associated with Ca²⁺-chelating. In this study, the IgE reactivity of Pacific mackerel Scomber japonicus parvalbumin (Sco j 1) was demonstrated to be markedly reduced (60–100% reduction) by Ca²⁺-depletion, similar to Cyp c 1. Three Sco j 1 mutants (D51A, D90A, D51/90A), with modifications in either one or both of the two Ca²⁺-binding sites, were then constructed by site-directed mutagenesis, followed by expression in Escherichia coli , and evaluated for their IgE reactivity. Interestingly, the double mutant (D51/90A), probably devoid of Ca²⁺-binding capacity, exhibited a significantly reduced IgE reactivity (equivalent to 0.0–7.5% of the IgE reactivity of natural Sco j 1). The results suggest that the IgE-binding ability of Sco j 1 largely depends on the solid conformation mediated by Ca²⁺-chelating, and that the hypoallergenic D51/90A will be a useful tool for the specific immunotherapy of fish allergy.     

Oolong tea theasinensins attenuate cyclooxygenase-2 expression in lipopolysaccharide (LPS)-activated mouse macrophages: structure-activity relationship and molecular mechanisms

Oolong tea theasinensins are a group of tea polyphenols different from green tea catechins and black tea theaflavins. The present study reports the inhibitory effects of oolong tea theasinensins on the expression of cyclooxygenase-2 (COX-2) and underlying molecular mechanisms in lipopolysaccharide (LPS)-activated murine macrophage RAW264 cells. The structure-activity data revealed that the galloyl moiety of theasinensins played an important role in the inhibitory actions. Theasinensin A, a more potent inhibitor, caused a dose-dependent inhibition of mRNA, protein, and promoter activity of COX-2. An electrophoretic mobility shift assay (EMSA) revealed that theasinensin A reduced the complex of NF-κB- and AP-1-DNA in the promoter of COX-2. Signaling analysis demonstrated that theasinensin A attenuated IκB-α degradation, nuclear p65 accumulation, and c-Jun phosphorylation. Furthermore, theasinensin A suppressed the phosphorylation of MAPKs, IκB kinase α/β (IKKα/β), and TGF-β activated kinase (TAK1). These data demonstrated that the down-regulation of TAK1-mediated MAPKs and NF-κB signaling pathways might be involved in the inhibition of COX-2 expression by theasinensin A. These findings provide the first molecular basis for the anti-inflammatory properties of oolong tea theasinensins.     

Real time detection of farm-level swine mycobacteriosis outbreak using time series modeling of the number of condemned intestines in abattoirs

Mycobacteriosis in swine is a common zoonosis found in abattoirs during meat inspections, and the veterinary authority is expected to inform the producer for corrective actions when an outbreak is detected. The expected value of the number of condemned carcasses due to mycobacteriosis therefore would be a useful threshold to detect an outbreak, and the present study aims to develop such an expected value through time series modeling. The model was developed using eight years of inspection data (2003 to 2010) obtained at 2 abattoirs of the Higashi-Mokoto Meat Inspection Center, Japan. The resulting model was validated by comparing the predicted time-dependent values for the subsequent 2 years with the actual data for 2 years between 2011 and 2012. For the modeling, at first, periodicities were checked using Fast Fourier Transformation, and the ensemble average profiles for weekly periodicities were calculated. An Auto-Regressive Integrated Moving Average (ARIMA) model was fitted to the residual of the ensemble average on the basis of minimum Akaike’s information criterion (AIC). The sum of the ARIMA model and the weekly ensemble average was regarded as the time-dependent expected value. During 2011 and 2012, the number of whole or partial condemned carcasses exceeded the 95% confidence interval of the predicted values 20 times. All of these events were associated with the slaughtering of pigs from three producers with the highest rate of condemnation due to mycobacteriosis.