Muscle fibre distribution in forelimb,hindlimb and trunk muscles in three bat species: The little Japanese horseshoe,greater horseshoe and Egyptian fruit

This study characterised muscle fibres in trunk, forelimb and hindlimb muscles of three bat species: little Japanese horseshoe (Rhinolophus cornutus), greater horseshoe (Rhinolophus ferrumequinum) and Egyptian fruit (Rousettus aegyptiacus). Twenty-seven muscles from trunk, forelimb and hindlimb were dissected, weighed and analysed by immunohistochemistry and sodium didecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and determined their cross-sectional areas (CSA). Results showed that Type IIa and Type IIa/x made the highest proportion of total muscle mass. Moderate proportion was formed by Type IIb. Type I and IIx appeared at very low levels in all bats. Type IIb was the only fibre type detected in patagial muscles in wing membrane of greater horseshoe while other fibre types were not observed. Type I muscle fibres were very few and appeared infrequently in fifteen muscles of Egyptian fruit and in only one muscle in each, greater horseshoe and little Japanese horseshoe. Type IIx was also detected in three muscles in greater horseshoe and only one muscle in Egyptian fruit but none in little Japanese horseshoe. The highest average CSA μm² was detected in Type IIb and values were 734.2μm² for LHB; 1537.9μm² for GHB and 1,720.9μm² for EFB. Lowest and undetermined values were observed for Type I and IIx. These data demonstrate that Type IIa, IIa/x and IIb form significant proportion of adult bat muscle mass and Type IIb is the largest fibre type. The distribution pattern is suggestive of specialised functions of the fibres in relation to orientation and speed of bats during flight.     

Databases for technical aspects of immunohistochemistry: 2021 update

With the aim of sharing information about the technical aspects of immunohistochemistry (IHC) and facilitating the selection of suitable antibodies for histopathological examination, this technical report describes the results of a questionnaire distributed during the period of 2018 to 2019 among members of the Conference on Experimental Animal Histopathology. Additionally, it describes the immunological properties and supplier details (clone, supplier, catalog number, species reactivity, etc.) as well as the IHC staining conditions (fixing solution, fixing time, embedding, antigen retrieval method, antibody dilution, incubation time, incubation temperature, positive control tissue, blocking condition, secondary antibody information, etc.) for a total of 509 primary antibodies (comprising 220 different types). These survey results were an update on the contents reported by CEAH in 2017.     

Effects of tea polyphenols on the invasion and matrix metalloproteinases activities of human fibrosarcoma HT1080 cells

The effects of tea polyphenols on the invasion of highly metastatic human fibrosarcoma HT1080 cells through a monolayer of human umbilical vein endothelial cells (HUVECs) and the accompanying basal membrane were investigated. Among the tea polyphenols tested, epicatechin gallate (ECg), epigallocatechin gallate (EGCg), and theaflavin strongly suppressed the invasion of HT1080 cells into the monolayer of HUVECs/gelatin membrane, whereas epicatechin, epigallocatechin, tea flavonols, tea flavones, and gallate derivatives had no effect. Both theaflavin-digallate and theasinensin D showed a weak invasion inhibitory effect. ECg significantly inhibited the invasion without cytotoxicity against cancer cells and HUVECs. Ester-type catechins (ECg and EGCg) and theaflavin strongly suppressed the gelatin degradation mediated by matrix metalloproteinase (MMP) 2 and MMP-9, which were secreted into the conditioned medium of HT1080 cells. In conclusion, among the tea polyphenols tested, ECg was considered to be the agent with the most potential antimetastasis activity because it inhibited invasion in the absence of cytotoxicity.     

Experimental studies on SO2 injuries in higher plants III: Inhibitory effect of suite ion on 14 CO 2 fixation

Photosynthesis decreases reversibly in plants exposed to SO₂. Photosynthesis recovers when the exposure to SO₂ is discontinued. Inactivation of a photosynthetic enzyme, ribulose-1,5-diphosphate carboxylase, by sulfonation of its SH groups was investigated as a cause of the reversible reduction of photosynthesis. The relationship between the sulfite ion concentration in the reaction mixture and ¹⁴CO₂ fixation catalized by the enzyme which was prepared from alfalfa leaves was explored by using radioactive NaHCO₃. About 50% and 85% inhibitions of ¹⁴CO₂ fixation were observed at 3 × 10^?3 M and 3 × 10^?2 M concentration of sulfite ion in the reaction mixture, respectively. The accumulation of 3 × 10^?4 M sulfite ion on the reaction site of the enzyme involved in the plants which were exposed to SO₂ could considerably reduce the CO₂ assimilation of the plant.     

Effects of dietary phytol on tissue accumulation of phytanic acid and pristanic acid and on the tissue lipid profiles in mice

Recent in vitro evidence suggests that the phytol‐derived fatty acids, phytanic acid (PA) and pristanic acid (PrA), are components of animal products with the potential to cause both beneficial and harmful effects on human health. In this study, we investigated the in vivo tissue accumulation of PA and PrA and the changes in tissue lipid profiles, using mice fed a phytol‐containing diet. After 4 weeks of treatment with a diet containing 1.0% phytol, plasma, adipose tissue, liver, and brain were collected and their lipid profiles were biochemically and gas‐chromatographically determined. Dietary phytol caused PA and PrA accumulation in the adipose tissue and liver but not in the brain, and reduced plasma and liver triacylglycerol levels. Phytol intake also decreased the fatty acid concentrations in the adipose tissue, especially polyunsaturated fatty acids such as linoleic acid, but increased the concentrations of these fatty acids in the liver. However, dietary phytol had a low impact on the brain lipid profile. This study suggests that dietary phytol intake caused accumulation of PA and PrA and modified lipid profiles in the adipose tissue and liver, but that the brain is an insusceptible tissue to dietary phytol‐induced changes.     

Effect of oral administration of a single bolus of six different protein sources on digestive physiology of red seabream Pagrus major juveniles

Fish Physiology and Biochemistry - To reveal direct effects of various protein sources on digestive physiology of red seabream, Pagrus major (38.5 ± 0.4 g), six different...     

A plastidic glucose-6-phosphate dehydrogenase is responsible for hypersensitive response cell death and reactive oxygen species production

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in the supply of reduced nicotine amide cofactors for resistance to biotic and abiotic stresses. Here, we show participation of the plastidic P2 isoform of G6PDH in plant immunity. A cytosolic isoform (NbG6PDH-Cyto) and two plastidic isoforms (NbG6PDH-P1 and NbG6PDH-P2) cloned from Nicotiana benthamiana were localized in cytosol and chloroplasts, respectively. Hypersensitive response (HR) cell death and NADPH oxidase (RBOH; respiratory burst oxidase homolog)-dependent reactive oxygen species (ROS) production after recognition of INF1 elicitin, secreted by oomycete Phytophthora infestans, decreased in NbG6PDH-P2-silenced plants, but not in NbG6PDH-Cyto- and NbG6PDH-P1-silenced plants. Silencing of the cytosolic NAD kinase NbNADK1, which phosphorylates NADH to form NADPH, compromised HR cell death and ROS production, and concomitant silencing with NbG6PDH-P2 reduced HR cell death and ROS to levels near those in NbG6PDH-P2-silenced plants. Similarly, silencing NbG6PDH-P2 and NbNADK1 resulted in high susceptibility to P. infestans. These results suggest that NADPH produced by the P2 isoform of G6PDH in chloroplasts is responsible for HR cell death and ROS production mediated by RBOH and that NbNADK1 is involved in this pathway.     

Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms

Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.     

Genetic variation of high-molecular-weight glutenin subunit composition in Asian wheat

To clarify the genetic properties of the HMW glutenin subunit composition of Asian endemic wheats, SDS–PAGE analysis was conducted using 1,139 bread wheat accessions that were originally collected in Asia. The samples were divided into six regional groups, Western Asia, Caucasia, Central Asia, Afghanistan, Southern Asia, and Eastern Asia. The genotype Glu-A1c, Glu-B1b, and Glu-D1a encoding subunits null, 7+8, and 2+12 had an overall frequency of 55.2%. Thus, we conclude that it is the typical genotype of the HMW glutenin subunits that characterize Asian endemic wheat. The frequency of the typical Asian genotype was relatively high in the central belt of Asia (Western Asia, Afghanistan, and Eastern Asia) and low in the marginal regions (Caucasia, Central Asia, and Southern Asia). In Southern Asia, the frequency of Glu-B1i, which encodes subunit 17+18, was the highest at the Glu-B1 locus. In Caucasia and Central Asia, the frequency of Glu-D1d, which encodes subunit 5+10 (which is considered to be the most useful for making bread), was high. The level of genetic variation, as estimated using the frequencies of the various alleles, was relatively low in the central belt of Asia and high in the marginal regions. Among the three Glu-1 loci, the highest number of alleles was detected at the Glu-D1 locus. This result was caused by the presence of rare Asian specific alleles at the Glu-D1 locus, in which a newly found allele, Glu-D1bs, encoding subunit 2.1+12 was included.     

Annual variation of natural 15N abundance in tea leaves and its practicality as an organic tea indicator

To obtain basic knowledge about the relationship between the application of organic fertilizers and the δ15N values of leaves of organically grown tea plants, annual variations in the δ15N values of the tea leaves were investigated. Although variations did not immediately arise after the application of organic fertilizers, differences in the δ15N values between organic and conventional cultivations appeared basically after three years from the beginning of the organic cultivation except when an organic fertilizer with a low δ15N value was applied, and the variation depended on the δ15N values of the fertilizers. In addition, the effectiveness of the δ15N values as a practical indicator of organic teas was examined. The tea leaves collected from organic farms did not always have higher δ15N values than the commercially available nonorganic teas. This result demonstrates that it is not easy to discriminate organic teas from nonorganic teas simply by their δ15N values.