A simple and specific assay of glycosyltransferase and glycosidase activities by an enzyme-linked immunosorbent assay method, and its application to assay of galactosyltransferase activity in sera from patients with cancer.

A simple, sensitive, and specific assay method for glycosyltransferase and glycosidase activities has been established by means of an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody, H-11 directed to lactoneotetraosylceramide (nLc4Cer). Enzyme activity was determined by assaying the amount of reaction product, nLc4Cer with the ELISA method. For the assay of galactosyltransferase activity, lactotriaosylceramide (Lc3Cer) immobilized on a 96-well microtiter plate was incubated with bovine milk galactosyltransferase in cacodylate buffer (pH 6.8) containing Triton CF-54, Mn2+, and UDP-galactose. Optimum incubation conditions for the enzyme were determined. Glycosidase activity was also assayed by the ELISA method by using Clostridium perfringens sialidase and neolacto-series gangliosides as substrates, and the substrate specificities towards the gangliosides were examined. By this method, 3-100 pmol of reaction product could be determined. The assay method has several advantages as follows: 1, the method is simple; 2, separation of the reaction product is not required; 3, quantification and identification of the reaction product were done simultaneously; 4, naturally occurring substrates are available (especially for glycosidase); 5, many samples can be assayed in one microplate; 6, sensitivity is very high. The present method was applied for the detection of galactosyltransferase in human sera. Significant elevations of the galactosyltransferase levels were observed in the sera from cancer patients. The formation of nLc4Cer was confirmed by employing the TLC-immunostaining method for bands of Lc3Cer after incubation of the bands with serum and cofactors on an HPTLC plate.     

Changes in coenzyme Q level in mitochondria of cirrhotic rat liver

In the cirrhotic rat liver induced by carbon tetrachloride and phenobarbitone, the concentrations of mitochondrial Coenzyme Q were measured in comparison with other respiratory components. The concentration of cytochrome a(+a3) and Coenzyme Q significantly increased in the cirrhotic liver, without any changes in the ratio of Coenzyme Q to cytochrome a(+a3). It is suggested that such increase of Coenzyme Q plays an important role as one of the adaptive responses to compensate for the prolonged metabolic overload on the mitochondrial respiratory assembly. Also, from the findings that the concentrations of cytochrome a(+a3) in the mitochondria of cirrhotic liver increase concomitant with the severity of cirrhosis, it is suggested that the rise of Coenzyme Q levels may be one of the indicators for the decreased functional reserve capacity in liver cirrhosis.     

In vitro ectomycorrhization of Tricholoma matsutake strains is differentially affected by soil type

The Japanese delicacy Tricholoma matsutake has been conducted in vitro ectomycorrhizal syntheses for more than 20 y. The development of its ectomycorrhizal structures varies among experimental systems. Here, we examined the effects of soil-fungus interactions on the early stage of in vitro T. matsutake ectomycorrhization. Axenic Pinus densiflora seedlings were transplanted into autoclaved natural inorganic soil, inoculated with the cultured mycelium of T. matsutake, and incubated for 90 d in vitro. Both soil type and fungal strain significantly affected host plant growth; host plant growth and mycorrhization levels significantly differed among soil type/fungal strain combinations. Therefore, the selection of T. matsutake strains for optimal mycorrhization must take into account such fungal and soil properties.     

Design,synthesis and evaluation of curcumin-based fluorescent probes to detect Aβ fibrils

Amyloid β fibrillation is an early event in Alzheimer’s disease, so its detection is important to understand its roles in Alzheimer’s disease. Curcumin, which has poor water solubility, has been reported to have many pharmacological activities including potent anti-amyloid β fibril activity in Alzheimer’s disease. In this study, we found that curcumin analogues with the fluorescence property instead of non-inhibition of amyloid β fibrils. The development of new curcumin analogue, Me-CUR (9), as fluorescent switchable probe to detect amyloid β fibrils is described. Me-CUR (9) shows excellent fluorescence, especially higher than ThT (4), in the presence of amyloid β fibrils. These results suggest that Me-CUR (9) can become a useful in vitro amyloid fluorescence sensor for diagnosis of Alzheimer’s disease.     

Analysis of ear side of mobile phone use in the general population of Japan

This study aimed to clarify the distribution of the ear side of mobile phone use in the general population of Japan and clarify what factors are associated with the ear side of mobile phone use. Children at elementary and junior high schools (n = 2,518) and adults aged ≥20 years (n = 1,529) completed an Internet‐based survey. Data were subjected to a logistic regression analysis. In children, due to the tendency to use the dominant hand, we analyzed the factors associated with the use of right ear in right‐handed people. Statistically significant differences were observed only in talk time per call (odds ratio (OR) = 2.17; 95% confidence interval (CI): 1.22–3.99). In adults, due to the tendency to use the left ear, we analyzed factors associated with the use of left ear in right‐handed people. Significant differences were observed in those aged 30–39 years (OR = 2.55; 95% CI: 1.79–3.68), those aged 40–49 years (OR = 3.08; 95% CI: 2.15–4.43), those aged >50 years (OR = 1.85; 95% CI: 1.20–2.85), and in those with a percentage of total talk time when using mobile phones at work of 51–100% (OR = 1.75; 95% CI: 1.21–2.55). We believe that future epidemiological studies on mobile phone use can be improved by considering the trends in mobile phone use identified in this study. Bioelectromagnetics. 39:53–59, 2018. © 2017 Wiley Periodicals, Inc.     

The SMXA: a new set of recombinant inbred strain of mice consisting of 26 substrains and their genetic profile

A new set of recombinant inbred (RI) strain SMXA consisting of 26 substrains was established between SM/J and A/J. The history of the SMXA RI strains and their genetic prolife covering 158 genetic marker loci are reported. From the strain distribution pattern among SMXA RI strains, the chromosomal location of salivary and tear protein genes Spel-r, Spel-s, Spe2, and Tpe1 were newly determined.     

Juveniles of two sillaginids,Sillago aeolus andS. sihama,occurring in a surf zone in the Philippines

Sillaginid juveniles collected from the surf zone at Tigbauan, Iloilo, Philippines, between May 1986 and September 1987 were identified asSillago aeolus (n=702, 8.9–26.0 mm SL) andS. sihama (n=3414, 8.6–22.9 mm SL), based on the numbers of dorsal and anal soft fin rays and vertebrae. The two species were easily distinguishable by the pattern of melanophores distributed on the caudal fin base,S. aeolus having a triangular-shaped cluster, whereas the melanophores formed a vertical line inS. sihama. The ratios of pre-anal and caudal peduncle lengths to SL also differed between the species, both being higher inS. aeolus. The occurrence ofS. aeolus was limited to the dry season, from January to March. On the other hand,S. sihama occurred year-round, although a peak was observed in the dry season, from November to April.     

Peg5/Neuronatin is an imprinted gene located on sub-distal chromosome 2 in the mouse.

We have established a systematic screen for imprinted genes using a subtraction-hybridization method with day 8.5 fertilized and parthenogenetic embryos. Two novel imprinted genes, Peg1/Mest and Peg3, were identified previously by this method, along with the two known imprinted genes, Igf2 and Snrpn. Recently three additional candidate imprinted genes, Peg5-7 , were detected and Peg5 is analyzed further in this study. The cDNA sequence of Peg5 is identical to Neuronatin, a gene recently reported to be expressed mainly in the brain. Two novel spliced forms were detected with some additional sequence in the middle of the known Neuronatin sequences. All alternatively spliced forms of Peg5 were expressed only from the paternal allele, confirmed using DNA polymorphism in a subinterspecific cross. Peg5/Neuronatin maps to sub-distal Chr 2, proximal to the previously established imprinted region where imprinted genes cause abnormal shape and behavior in neonates.     

Function and structure of leaves contributing to increasing water storage with height in the tallest <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> trees of Japan

Key message

In Cryptomeria japonica , transfusion tissue in leaves may have functions of water storage and supply, which could compensate for hydraulic constraints with increasing height.

Abstract

The tallest trees of Cryptomeria japonica occur in climatic regions similar to the world’s tallest trees. We hypothesized that tall C. japonica trees would have evolved adaptive mechanisms to overcome height growth limitation. Here, we focused on foliar water storage, a mechanism recently discovered in Sequoia sempervirens. In C. japonica, leaf water potential at turgor loss did not change with height or light availability, while leaf hydraulic capacitance and succulence (water content per leaf surface area) increased, suggesting hydraulic compensation. Plasticity of leaf morphology could contribute to avoiding negative effects of height on photosynthesis. We also focused on the structure and function of transfusion tissue in leaves and its role in water storage and supply. Cross-sectional area of transfusion tissue increased with height, whereas that of xylem was constant. We confirmed that water flowed from vascular bundle to mesophyll via the transfusion tissue. Cryo-scanning electron microscopy images of leaf cross sections showed that transfusion cells were flattened, but not fully dehydrated when leaf water potential decreased in situ and by experimental dehydration, and cell deformation was more marked for treetop leaves than for lower-crown leaves. The shape of transfusion cells recovered at predawn as well as after experimental rehydration. As in S. sempervirens, transfusion tissue of C. japonica may function as a hydraulic buffer, absorbing and releasing water according to leaf water status. Anatomical and hydraulic properties contributing to foliar water storage may be an adaptive mechanism acquired by tall Cupressaceae trees to overcome the hydraulic constraints on physiological function with increasing height.