Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.     

Effects of whey protein hydrolysate on growth promotion and immunomodulation in mouse pups in artificial rearing system

This study aimed to investigate the effects of whey protein hydrolysate (WPH) on the growth and immunity of mouse pups in artificial rearing (AR) system. Mouse pups were reared in the AR system with artificial milk including 5% WPH (AR with WPH) or not (AR without WPH), and the remaining pups were reared by their mother (dam) for 14 days after birth. The body weight change and body weight gain rates in the AR with WPH group were significantly higher than those observed in the AR without WPH group and similar to those in the dam group. Moreover the feed and protein efficiencies in the AR with WPH group were significantly higher than those of the AR without WPH group. In addition, the supplement of WPH in the AR system was shown to significantly elevate the number of CD3⁺CD8⁺, B220⁺CD19⁺, IA/IE⁺CD11c⁺, and CD11b⁺ in the thymocyte and/or splenocyte, and the thymus weight. Furthermore, MALDI‐TOF/MS analysis identified the amino acid sequences corresponding to some peptides, and indicated that VRTPEVDDE had the highest relative intensity among the peptides from tested WPH. Therefore, WPH would be required to not only promote growth, but also exert immunomodulatory activities in mouse pups in AR system.     

Exposure to blue LED light before the onset of darkness under a long‐day photoperiod alters melatonin secretion,feeding behaviour and growth in female dairy calves

The effect of blue LED on melatonin secretion, feeding behaviour and growth was addressed in Holstein female dairy calves. In Exp.1, six animals (8 weeks old, 97 ± 4.1 kg BW) were exposed to yellow or blue LED for 2 hr before darkness over 7 days under a long‐day photoperiod (LDPP). In Exp. 2, six animals (8 weeks old, 88.5 ± 4.8 kg BW) were exposed to blue light from a white LED all daytime or a yellow LED for 2 hr before the darkness of LDPP (blue light cut) over 3 weeks. In Exp. 1, blue light mildly suppressed melatonin secretion during the 2‐hr treatment but did not affect the timing of the nightly melatonin rise. However, the rise in nighty melatonin levels was higher with yellow than blue LED. In Exp. 2, white LED completely suppressed melatonin secretion during the 2‐hr treatment, but plasma melatonin concentrations were similar during the darkness. Grass hay intake, rumination time, frequency of water intake and body weight gain were higher in animals exposed to the yellow rather than the white LED. Overall results indicate that exposure to blue light from white LEDs under an LDPP suppresses melatonin secretion and might negatively impact the development of female dairy calves.     

Outbreak of human infections with uncommon Salmonella serotypes linked to pet bearded dragons, 2012–2014

Reptiles are one of the fastest growing sectors in the United States pet industry. Reptile‐associated salmonellosis (RAS) continues to be an important public health problem, especially among children. We investigated an outbreak of human Salmonella infections resulting from serotypes Cotham and Kisarawe, predominately occurring among children. An outbreak of illnesses was identified in persons with exposure to pet bearded dragon lizards. Human and animal health officials, in cooperation with the pet industry, conducted epidemiologic, traceback and laboratory investigations. Onsite sampling was conducted at two US breeding facilities, one foreign breeding facility, and a large pet retail chain. A total of 166 patients in 36 states were identified with illness onset dates from 02/2012–06/2014. The median patient age was 3 years (range, <1–79 years), 57% were aged ≤5 years, and 37% were aged ≤1 year. Forty‐four patients (37%) were hospitalized, predominantly children. Sampling at breeding facilities and a national pet store chain resulted in isolation of outbreak serotypes at each facility; isolation proportions ranged from 2%–24% of samples collected at each facility.Epidemiologic, microbiologic and traceback evidence linked an outbreak of uncommon Salmonella serotypes to contact with pet bearded dragons. The high proportion of infants involved in this outbreak highlights the need to educate owners about the risk of RAS in children and the potential for household contamination by pet reptiles or their habitats. Strategies should be developed to improve breeding practices, biosecurity and monitoring protocols to reduce Salmonella in the pet reptile trade.     

Wood density and carbon and nitrogen concentrations in deadwood of Chamaecyparis obtusa and Cryptomeria japonica

Estimating carbon (C) and nitrogen (N) stocks in deadwood in forests nationwide is required for understanding large-scale C and N cycling. To do so requires estimated values of wood density and C and N concentrations. Additionally, parameters that show variation should be examined. In this study, we clarified the estimated values and the variation in three parameters in each decay class of each of two tree species and examined whether dead log diameter and region contribute to variation in the parameters. Data were collected from 73 Chamaecyparis obtusa (Sieb. et Zucc.) Endl. plantations and 66 Cryptomeria japonica D. Don plantations throughout Japan. Wood densities decreased from 386 to 188?kg?m^?3 for C. obtusa and from 334 to 188?kg?m^?3 for C. japonica in decay classes 1–4. The variation in wood density increased with decay class, and the coefficient of variance increased from 13.9% to 46.4% for C. obtusa and from 15.2% to 48.1% for C. japonica. The N concentrations increased from 1.04 to 4.40?g?kg^?1 for C. obtusa and from 1.11 to 2.97?g?kg^?1 for C. japonica in decay classes 1–4. The variation in N concentration increased with decay class, and the coefficient of variance increased from 51.9% to 76.7% for C. obtusa and from 50.3% to 70.4% for C. japonica. Log diameter and region contributed to variations in wood density and N concentration in decay classes 1 and 2 for C. obtusa and C. japonica. However, no relationship was observed between regional climates and the two parameters. In contrast, C concentrations ranged from 507 to 535?g?kg^?1 and were stable with much lower coefficients of variance throughout the decay classes for both tree species. Thus, we recommend that the same C concentration can be adapted for all decay classes of both tree species.     

Chemical and morphological changes of porous hydrated calcium silicate in paddy soil

The chemical and morphological changes of porous hydrated calcium silicate material (PS) during the dissolution process in paddy soil were investigated by using both a scanning electron microscope (SEM) and an energy dispersive X-ray (EDX) analysis. The results showed that original PS consisted of agglomerate of various sizes with almost the same elemental composition. The SEM images at a high magnification showed that the original PS consisted of agglomerates of small platelike crystals (≦ μm) of tobermorite [Ca₅(Si₆O₁₈-H₂)·4H₂O]. On the other hand, elemental composition showed that the PS agglomerates in paddy soil altered to Si- and Ca-rich agglomerates after incubation for 53 d. The morphological differences between the two types of agglomerates were observed by SEM at a high magnification. The Si-rich agglomerates were similar to the original PS in morphology, whereas the Ca-rich agglomerate appeared as a large crystal. The X-ray diffraction patterns obtained in a previous study suggested the presence of quartz and calcite in PS after incubation for 53 d (Saigusa et al. 2000: Soil Sci. Plant Nutr., 46, 89–95). Si- and Ca-rich agglomerates were identified as silica skeletons of PS and calcite, respectively. It was suggested that the silica skeletons of PS remained as a silicon source for rice plants even after the disappearance of tobermorite revealed by the X-ray diffractogram.     

Phospholipid fatty acid composition of microbiota in the percolating water from a rice paddy microcosm

The microbiota in the percolating water from the plow layer soil in paddy fields was studied based on the composition of phospholipid fatty acids (PLFAs) in a pot experiment. The mean concentrations of PLFAs in the percolating water were 17±5 and 11±4 µg L^-1 in the planted and non-planted pots, respectively. The dominant PLFAs in the percolating water were 16: 0, 16: 1ω7c, 18: 1ω7, 18: 1ω9, il5: 0, and ail5: 0 PLFAs in both the planted and non-planted pots. The dominance percentage of 18: 3ω6c and 17: 1ω8 PLFAs increased at the late stage of rice growth in the planted pots. The percolating water from the planted pots contained in a higher percentage of straight mono-unsaturated PLFAs and a lower percentage of branched-chain PLFAs than that from the non-planted pots. Considerable differences in the PLFA composition in the percolating water were observed between the planted and non-planted treatments and with the duration of the growth period. Principal component analysis indicated that the microbiota in the percolating water was derived from the microbiota in the floodwater and in the plow layer soil. Cluster analysis showed that the similarity of the PLFA composition in the percolating water to the PLFA composition in the plow layer soil was higher than that in the floodwater. The stress factor that was estimated from the trans/cis ratio of 16: 1ω7 PLFA was 0.08±0.04 and 0.14±0.05 in the percolating water from the planted and non-planted pots, respectively, which indicated that the degree of stress on the microbiota in the percolating water from the planted pots was low in a similar way to the degree of stress on the microbiota in the floodwater, while the degree in the percolating water from the non-planted pots was similar to that in the plow layer soil, respectively.     

Responses of antioxidant enzymes in the needles of hinoki cypress (Chamaecyparis obtusa) seedlings to nutrient solutions containing various calcium/aluminum ratios

An attempt was made to examine the possible connection between the various ratios of calcium/aluminum (Ca/Al) in the nutrient solution of plant cultures and the active oxygen scavenging system of hinoki cypress (Chamaecyparis obtusa) seedlings. The hinoki cypress seedlings were transferred to nutrient solutions containing 5 mM AlCl₃ together with various concentrations of Ca(NO₃)₂ in pots containing glass beads and Teflon tips. The activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) in the needles were estimated at several stages during the 7-day treatment. The samples treated with the lower Ca/Al solutions show the highest SOD activities. The activities of APX and GR, both of them involved in the ascorbate-glutathione cycle, showed the same tendency with decrease to higher Ca/Al ratio. These results indicate that rhizospheric Ca might compete with Al and ameliorate Al toxicity on and in the roots, the Al stress is not transformed to the needles after a few days, and the ascorbate-glutathione cycle in the hinoki cypress needles might fluctuate and be suppressed by the rhizospheric Al stress during the 7 days. This work was supported in part by funding from the Japan Science & Technology Corporation, the CREST program 1996–2001, and the Center for Forest Decline Studies.     

Reactivity of green tea catechins with formaldehyde

In the reaction of green tea catechins with formaldehyde at room temperature (25°C), tea catechins were found to have reactivity. In particular, (-)-epicatechin gallate and (-)-epigallocatechin gallate, which have a galloyl moiety at the C-3 position, showed higher reactivity than (+)-catechin, (-)-epicatechin, or (-)-epigallocatechin. Reactivity of various kinds of simple phenolic compounds and flavonoids with formaldehyde was also examined. Among these compounds, only phloroglucinol showed reactivity to the same degree as that of nongalloylated catechins. These results suggest that factors for reactivity with formaldehyde at room temperature may be the presence of a phloroglucinolic A-ring structure and the absence of the electron-attractive group such as a carbonyl group in Cring. The comparison of the reactivity of 3-O-acylated catechins with that of 3-O-galloylated catechins indicated that only a galloyl group effectively enhanced reactivity with formaldehyde.     

Autophagy is essential for preimplantation development of mouse embryos

After fertilization, maternal proteins in oocytes are degraded and new proteins encoded by the zygotic genome are synthesized. We found that autophagy, a process for the degradation of cytoplasmic constituents in the lysosome, plays a critical role during this period. Autophagy was triggered by fertilization and up-regulated in early mouse embryos. Autophagy-defective oocytes derived from oocyte-specific Atg5 (autophagy-related 5) knockout mice failed to develop beyond the four- and eight-cell stages if they were fertilized by Atg5-null sperm, but could develop if they were fertilized by wild-type sperm. Protein synthesis rates were reduced in the autophagy-null embryos. Thus, autophagic degradation within early embryos is essential for preimplantation development in mammals.