Neuroanatomical distribution of disease-associated prion protein in cases of bovine spongiform encephalopathy detected by fallen stock surveillance in Japan

Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disorder of cattle characterized by accumulation of the disease-associated prion protein (PrP(Sc)) in the central nervous system (CNS). The immunohistochemical patterns and distribution of PrP(Sc) were investigated in the CNS, brains, and spinal cords of 7 naturally occurring BSE cases confirmed by the fallen stock surveillance program in Japan. No animals showed characteristic clinical signs of the disease. Coronal slices of 14 different brain areas in each case were immunohistochemically analyzed using an anti-prion protein antibody. Immunolabeled PrP(Sc) deposition was widely observed throughout each brain and spinal cord. Intense PrP(Sc) deposition was greater in the thalamus, brainstem, and spinal cord of the gray matter than in the neocortices. The topographical distribution pattern and severity of PrP(Sc) accumulation were mapped and plotted as immunohistochemical profiles of the different brain areas along the caudal-rostral axis of the brain. The distribution pattern and severity of the immunolabeled PrP(Sc) in the CNS were almost the same among the 7 cases analyzed, suggesting that the naturally occurring cases in this study were at the preclinical stage of the disease. Immunohistochemical mapping of the PrP(Sc) deposits will be used to clarify the different stages of BSE in cattle.     

Approachability and contact behavior of commercial dairy calves to humans

Calves (n = 106) on four dairy farms were observed for their approachability to humans. All calves experienced similar rearing conditions: Beginning individual pen, after birth until weaning at about 2 months, where they were housed individually and fed milk and a milk replacement; Late individual pen, after weaning until grouping at about 3.5 months, where they were housed individually and fed hay, silage and concentrate feed; Beginning group pen, after grouping until 5 months, where they were housed in groups of 2–5 animals and fed hay, silage and concentrate feed; later group pen, from 5 to 7 months. The number of calves that contacted an experimenter who stood in front of their pens for 10 min was recorded on 6 separate days over 3 months. Latency to touch and time spent in activities during touching such as sucking, licking, biting and rubbing were also measured. There were no significant differences in the latency to touch and the ratio of touch to non‐touch calves between the rearing conditions and the farms. The time spent touching was significantly affected by the interaction between the rearing condition and the farm (P < 0.01). In detail, the time spent sucking (P < 0.001) and licking (P < 0.01) was different between the rearing condition × farm variables. The proportion of calves that approached and touched the experimenter tended to be higher in the farms in which a stockperson worked longer inside and outside their pens (both ρ = 0.95, P = 0.051). These results were interpreted according to the perspectives of early positive reinforcement with food and the habituation process to humans existing nearby.     

Sex and age differences in meat composition of Yeso sika deer (Cervus nippon yesoensis) reared for a short period after capture in the wild

Yeso sika deer captured in winter around Lake Akan in Hokkaido were reared for 8–10 months at Tokyo University of Agriculture in Abashiri. Six 1‐year‐old females and males and six 2‐year‐old or older (adult) females and males were slaughtered and their carcasses were processed. The chemical composition, mineral contents and fatty acid composition of the loin were measured. No marked influence of gender or age was noted in the chemical composition of loin. In the mineral contents, significant differences were noted. The potassium and sulfur contents were lower and the sodium content was higher in adult deer meat (P < 0.05, respectively) and the potassium content was higher in male deer meat (P < 0.05). Arsenic, cadmium or lead were not detected. In the unsaturated fatty acid, a significant interaction was detected (P < 0.05), and it was high in 1‐year‐old female deer meat and low in 1‐year‐old male deer meat. Significant gender or age differences were noted only in the mineral contents in the loin of deer reared for a short period after capture.     

New Terpenes from the Egyptian Soft Coral Sarcophyton ehrenbergi

Chemical investigations of the Egyptian soft coral Sarcophyton ehrenbergi have led to the isolation of compounds 1–3 as well as the previously reported marine cembranoid diterpene sarcophine (4). Structures were elucidated by comprehensive NMR and HRMS experimentation. Isolated compounds were in vitro assayed for cytotoxic activity against human hepatocarcinoma (HepG2) and breast adenocarcinoma (MCF-7) cell lines.     

Fuzzy neural network model for habitat prediction and HEP for habitat quality estimation focusing on Japanese medaka (Oryzias latipes) in agricultural canals

In the field of agriculture, development of evaluation techniques for environmental changes is urgently required for the purpose of finding a balance between growth in agricultural productivity and environmental considerations. The habitat evaluation procedures (HEP) constitute one technique for habitat assessment. While HEP is widely applied to estimate both habitat quality and quantity in an environment, it appears to be necessary to develop an accurate habitat prediction model in order to evaluate environments precisely. In fact, habitat selection by fish is affected by complicated interaction between multiple environmental factors, which makes it difficult to relate physical environments to habitat preference. In the present study, we utilize artificial neural networks (ANNs), which are commonly applied to model complex systems, to predict the habitat selection of Japanese medaka (Oryzias latipes) in agricultural canals. Considering the essential vagueness of fish behavior, fuzzy membership functions are introduced into the input layer, which advances ANN to fuzzy neural network (FNN). In addition, symmetric triangular fuzzy numbers are employed to account for uncertainty in measurement errors and dispersions of physical environment. The FNN model precisely predicts the habitat preference of Japanese medaka in an agricultural canal, and the results show a good agreement between the calculated and observed habitat suitability indices (HSI). Finally, the habitat quality of two different reaches at the same point in time is compared using HEP, with a view of suitable habitat for Japanese medaka.     

Comparison of the early response to aluminum stress between tolerant and sensitive wheat cultivars: Root growth,aluminum content and efflux of K+

In order to characterize the mechanism of Al tolerance (Atlas 66) and Al sensitivity (Scout 66) in two cultivars of wheat (Triticum aestivum L.), the early responses to Al stress under acidic conditions were investigated. Marked inhibition of root elongation of Scout was observed upon treatment with 10 μM AlCl₃ for less than 3 h. The inhibition of root elongation of Scout was reversed within 3 days when the treated samples were transferred to a solution without Al. However, treatment for 6 h with AlCl₃ repressed root elongation almost completely and irreversibly. Root elongation of Atlas was only partially inhibited by the treatment with 10 μM AlCl₃ for more than 6 h. Levels of Al in two portions of roots, namely, portions 0–5 mm and 5–10 mm from the tip, were lower in Atlas than those in Scout. In Atlas the levels of Al on a fresh weight basis in both portions were very similar, while the level of Al in the portion 0–5 mm from the tip was almost double than that in the 5–10 mm portion in Scout. A distinct increase in levels of Al in the 0–5 mm portion over that in the 5–10 mm portion of Scout was observed even after 3 h of treatment with AlCl₃.

Both Atlas and Scout were preloaded with K⁺ at pH 5.5 and transferred to distilled water at various pH values to monitor the efflux of K⁺. A reduction in the pH induced increases in the efflux of K⁺ in both cultivars, and the rate of efflux in Scout was twice that in Atlas at pH 4.2. AlCl₃ at concentrations as low as 5 μM markedly repressed K⁺efflux at pH 4.2 and this effect was more pronounced in Scout. Ca²⁺ also had a repressive effect on K⁺ efflux, while EGTA increased K⁺ efflux. Vanadate increased K⁺ efflux, a result that suggests the involvement of a H⁺ pump in K⁺ efflux. Ca²+ failed to repress the increased efflux of K⁺ caused by vanadate while Al repressed the K⁺ efflux even in the presence of vanadate. These results suggest that a low extracellular pH may cause an increase in the cytoplasmic concentration of H⁺ that is followed by depolarization of the plasma membrane, which may be modified by the efflux of K⁺ and H⁺. The characteristic difference in terms of K⁺ efflux between Atlas and Scout at low pH may be caused by differences associated with plasma membrane potentials, as follows. The net influx of H+ at low pH, which causes depolarization of the plasma membrane, is higher in Scout than in Atlas. The difference in the net influx of H⁺ may be regulated in part by Ca2⁺, that either repress the influx of H⁺ or the activate of the H⁺ pump. Inhibition of K⁺ efflux by Al, which tends to depolarize the plasma membrane at low pH, may be an important factor in determining sensitivity and/or tolerance to Al.     

Enhancement of nitrate efflux and the plasma membrane hydrogen ion‐transport activity of barley roots by nitrate treatment

The effect of nitrate (NO₃) on the activities of hydrogen (H⁺)‐ATPase and H⁺‐transport in the plasma membrane vesicles isolated from barley (Hordeum vulgare L. cv. Kikaihadaka) roots was investigated. After treatment with 10 mM NO₃ for longer than 24 h, both activities of H⁺‐ATPase and H⁺‐transport assayed in the presence of chloride (Cl) were stimulated less than 20%. While H⁺‐transport activity was approximately two times higher in NO₃‐grown roots than in control roots when assayed in the presence of NO₃ instead of Cl. This result suggests that NO₃ is permeable in the plasma membrane vesicles isolated from NO₃‐grown roots rather than that of the control, and thereby stimulate H⁺‐transport activity through the collapse of positive potentials established by H⁺‐ATPase. Furthermore, NO₃ efflux from roots was markedly enhanced after 48 h of the exposure to NO₃. Taken together, NO₃ efflux may be mediated by the permeability to NO₃ which is induced by NO₃.     

Quantitative analysis of the initial transport of fixed nitrogen in nodulated soybean plants using 15N as a tracer

The quantitative analysis of the initial transport of fixed isotope 15-nitrogen (¹⁵N) in intact nodulated soybean plants (Glycine max [L.] Merr. cv. Williams) was investigated at the vegetative stage (36 days after planting, DAP) and pod-filling stage (91 DAP) by the ¹⁵N pulse-chase experiment. The nodulated roots were exposed to N₂ gas labeled with a stable isotope ¹⁵N for 1 h, followed by 0, 1, 3 and 7 h of exposure with normal air. Plant roots and shoots were separated into three sections (basal, middle and distal parts) with the same length of the main stem or primary root. Approximately 80 and 92% of fixed N was distributed in the basal part of the nodulated roots at the vegetative and pod-filling stages by the end of 1 h of ¹⁵N₂ exposure, respectively. In addition, about 90% of fixed ¹⁵N was retained in the nodules and 10% was exported to root and shoot after 1 h of ¹⁵N₂ exposure at 91 DAP. The percentage distribution of ¹⁵N in the nodules at the pod-filling stage decreased from 90% to 7% during the 7 h of the chase period, and increased in the roots (14%), stems (54%), leaves (12%), pods (10%) and seeds (4%). The ¹⁵N distribution was negligible in the distal root segment, suggesting that N fixation activity was negligible and recycling fixed N from the shoot to the roots was very low in the initially short time of the experiment.     

Energy-dispersive x-ray analysis of phytin globoids in aleurone particles of developing rice grains

Abstract

In this study energy-dispersive X-ray analysis was used to discover changes in the composition of phytin globoids in aleurone particles of developing rice grains.

At early milky stage (the 7th to 10th day after flowering) many aleurone particles were observed as electron lucent vacuole-like particles in aleurone cells, some of which contained a small electron-dense inclusion (phytin globoid). The major mineral elements present in phytin globoids were phosphorus (P) and magnesium (Mg). Potassium (K) was also detectable but its concentration was extremely low relative to these two. Calcium (Ca) and zinc (Zn) were found as minor components. At early dough stage (the 17th to 19th day after ftowering) P, Mg and K were observed as the major mineral elements. The composition of mineral elements in phytin globoids tend to be constant in the late staae of ripening. On the other hand, minor elements, i.e. Ca and Zn, were only detectable in the early stage of ripening, suuesting that these elements accumulated in the aleurone particles only during the early stage, and in later stage their accumulation was either complete or at very low levels compared with those of K and Mg. The relative amount of Mg existing in phytin globoid remained roughly constant throughout the ripening periods, while that of K varied.

Energy-dispersive X-ray analysis of phytin globoids in developing rice aleurone particles confirmed that the accumulation of P, Mg and K in rice grains was closely related to the formation of phytin globoids. This analysis suggested further that the mechanism of accumulation of P into aleurone particles was very similar to that of Mg, while that of K differed from both. The results obtained suggested that Ca and Zn might be required in the formation of phytin globoid at the early ripening stage. EDX analysis of a protein body in the starchy endosperm revealed that there was no significant element detectable in an EDX spectrum other than sulfur. This suggests that lamellar concentric structures in protein bodies are not due to phytin.