Overexpression of the Barley Nicotianamine Synthase Gene HvNAS1 Increases Iron and Zinc Concentrations in Rice Grains

In humans, iron (Fe) and zinc (Zn) deficiencies result in major worldwide health problems. Transgenic technologies to produce Fe- and Zn-biofortified rice varieties offer a promising potential solution. Nicotianamine, the precursor of phytosiderophores, chelates Fe²⁺ and Zn²⁺ and plays an important role in transporting these metals to both vegetative and reproductive organs within the plant. Our objective was to increase Fe and Zn contents in rice grains by overexpressing the barley nicotianamine synthase gene HvNAS1. HvNAS1-overexpressing transgenic rice showed increased HvNAS1 expression and subsequent increases in endogenous nicotianamine and phytosiderophore content in shoots, roots, and seeds. Fe and Zn concentrations in polished T₁ seeds from transgenic plants increased more than three and twofold, respectively; Fe and Zn concentrations also increased in both polished and brown T₂ seeds. These results suggest that the overproduction of nicotianamine enhances the translocation of Fe and Zn into rice grains.     

Establishment of an Invasive Prostate Cancer Model in Transgenic Rats by Intermittent Testosterone Administration

We have established a transgenic rat for adenocarcinoma of the prostate (TRAP) model that features uniform adenocarcinoma development in prostatic lobes at high incidence within a short experimental period. However, no invasive carcinomas with reactive stroma characteristics similar to those in man were observed. We therefore have focused on a new model for invasive carcinoma of the prostate using TRAP rats. In experiment 1, male TRAP rats in groups 1 and 2 were treated with orchiectomy at day 0 of the experiment. Rats in groups 1–3 underwent testosterone propionate (TP) implantation from weeks 1 to 4 and from weeks 6 to 16. Rats in groups 1 and 3 were given 3,2’-dimethyl-4-aminobiphenyl (DMAB) after TP implantation. The rats of group 4 served as controls. In experiment 2, the rats were divided into three groups, none of which received DMAB or orchiectomy, treated with TP continuously or with the treatment withdrawn once or twice. In experiment 1, invasive adenocarcinomas with abundant collagenous stroma were found in the dorsolateral and anterior prostate, some of which showed perineural space invasion at week 16. The number of invasive carcinoma foci was most frequent in group 3. In experiment 2, invasive adenocarcinoma development in the lateral prostates was correlated with the number of TP administration/withdrawal cycles. In conclusion, our newly established rat model for invasive adenocarcinoma of the prostate could serve as a useful preclinical model for evaluating the in vivo efficacy of preventive and therapeutic agents targeting of the tumor microenvironment.     

A trial of intrauterine insemination using a fiberscope in the giant panda (Ailuropoda melanoleuca)

An intrauterine insemination technique using a fiberscope was investigated in the giant panda. A septal wall was present about 5 cm from the vulva, the external urethral orifice was present in the ventral region, and the entrance (pseudocervix) to the vagina was present in the dorsal region. The uterovaginal region protruded in the dorsal region about 15 cm from the pseudocervix. The external uterine orifice was present in the uterovaginal region, revealing that intrauterine insemination can be easily performed. This technique may greatly contribute to artificial reproduction of the giant panda.     

Nitrogen, phosphorus and potassium utilization and their cycling in a beef-forage production system

The objective of this study was to quantify nitrogen (N), phosphorus (P) and potassium (K) utilization and their cycling in a beef-forage production system. A survey was conducted at the Kyoto University experimental farm in Kyoto Prefecture, Japan, and included 9.5 ha of meadow and about 160 head of beef cattle. Forages cultivated on the meadow were ensiled and offered to breeding cows. The whole farm was regarded as a beef-forage production system. The system consisted of three components: animals, manure and soil/crop. Nutrient inputs, flows through the soil-plant-animal pathway and outputs were quantified for 2 periods for the whole farm. Nutrient utilization was evaluated by nutrient balances, utilization efficiencies (ratio of nutrient export to nutrient input) and cycling index. The average nutrient balances of N, P and K for the whole farm (kg/year/ha) were 435.7, 136.3, and 262.8, and the average nutrient utilization efficiencies were 0.08, 0.06, and 0.01, respectively. The characteristics of nutrient balances and utilization efficiencies for each component differed among the nutrients. The average cycling indices for N, P and K were 0.19, 0.17 and 0.49, respectively. This study illustrates the importance of internal nutrient cycling for improving nutrient utilization in beef production systems.     

Effects of structure on radical-scavenging abilities and antioxidative activities of tea polyphenols: NMR analytical approach using 1,1-diphenyl-2-picrylhydrazyl radicals

Tea (Camellia sinensis) leaves contain various antioxidants such as ascorbic acid (1) and polyphenols. This study tries to clarify the molecular mechanisms underlying the antioxidative and radical-scavenging activities of these antioxidants, and the reactivities of each antioxidant have been compared against that of the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH, 2) using nuclear magnetic resonance (NMR) analysis. Catechol (3) and (+)-taxifolin (4) were oxidized to o-quinone by 2. However, ethyl protocatechuate (5) and quercetin (6) were not oxidized to o-quinone, even though they possess a catechol structure. The radical-scavenging ability of o-dihydroxyl phenolic compounds with a conjugated olefinic double bond (e.g., 6) was superior to that of compounds without this bond (e.g., 4), whereas the ability of o-dihydroxyl phenolic compounds possessing a conjugated carbonyl bond (5) was inferior to that of compounds lacking this bond (3). Vicinal trihydroxyl phenolic compounds with a conjugated olefinic double bond [e.g., myricetin (7)] had an inferior scavenging ability as compared with compounds lacking this bond [e.g., pyrogallol (8)], but 7 was a better scavenger than compounds with a conjugated carbonyl double bond [e.g., ethyl gallate (9)]. In addition, vicinal trihydroxyl phenolic compounds (e.g., 9) were superior to o-dihydroxyl phenolic compounds (e.g., 6). Finally, 1 scavenged radicals more quickly than 8.     

Extraction of soybean oil from single cells

Single cells prepared from autoclaved soybeans and cellulase treatment of the cells were effective in digesting the cell walls of and extracting the oil from soybeans. The first cell wall of the soybean single cell was completely removed using cellulases; the thin and transparent second cell wall of the cell was swollen. Oil in the cell formed spherical or hemispherical oil drops, and oil leaking from the oil bodies was observed. The oil was almost retained within the second cell wall. Water-extractable substances were obtained at approximately >60% of the weight. Flotation of oil drops by centrifugation was easily done. Ambient n-hexane extraction was also possible; however, residual oil remained in the oil bodies. Protease or peptidase digested the structure of the oil bodies; however, separation of the oil and the hydrolysates was impossible. The oil from the oil bodies was obtained effectively (>85%) by pressing the single cells and/or cellulase-treated single cells.     

Cleaning procedures and cleanliness assessments of bucket milkers and suckling buckets on Japanese dairy farms

Cleanliness of milking equipment is known to be important for the safety of dairy products and to prevent the spread of diseases among cows. We investigated the cleaning procedures of milking equipment and suckling equipment on Japanese dairy farms, and the cleanliness of bucket milkers, suckling buckets, milk receivers, and bulk tanks, using adenosine triphosphate (ATP) bioluminescence test. Bulk tanks (except one bulk tank) and milk receivers were washed by automated cleaning, but all bucket milkers and suckling buckets were washed by manual cleaning. Detergents were often not used to clean bucket milkers and suckling buckets. The log₁₀ transformed relative luminescence units (LRLU) of equipment washed by manual cleaning was higher than equipment washed by automated cleaning. Clean surfaces (≤2.2 LRLU) were only observed on the bulk tank and the milk receiver. More than 50% of the LRLU of the mouthpiece, the rubber packing of claw, and the nipple of the suckling bucket were determined dirty. These results suggest that the cleanliness of the bucket milkers and the suckling buckets washed by manual cleaning was lower than that of the equipment washed by automated cleaning, and may be due to insufficient cleaning procedures.     

Swim bladder tumor in the young adult scoliotic medaka (Oryzias latipes)

A swim bladder tumor was detected in one scoliotic medaka aged 22 weeks. The tumor was located in the dorsal abdominal cavity, with maximum dimension of 1,850 × 1,500 µm. No swim bladder lumen was identified, and the region where the swim bladder lumen would have been located, was replaced with adipose tissues. The tumor was a non-invasive, expansile, and encapsulated solid mass with a few cysts, and comprised a homogenous population of well-differentiated, densely packed, gas glandular epithelium-like cells. The tumor mass was connected to a rete mirabile that showed a hyperplastic capillary plexus; however, the tumor cells did not invade the rete mirabile, thereby revealing that the tumor was an adenoma originating from the gas glandular epithelium of the swim bladder. Since proliferative lesions in the swim bladder have been reported in some teleosts with skeletal deformations, including medaka, the occurrence of a spontaneous swim bladder tumor in teleosts is considered to be closely associated with various types of skeletal deformation, and spinal curvature in particular.     

Estimation of the proportions of offspring having soft fruit and juicy fruit as evaluated by sensory tests in Japanese persimmon

Firmness and juiciness are important fruit attributes targeted in the breeding of Japanese persimmon (Diospyros kaki Thunb.). To facilitate the efficient production of cultivars with soft and juicy fruit, we successfully used the mid-parental values (MP) of sensory evaluations as independent variables in logistic regression models to estimate the proportions of offspring from a given cross that would have soft fruit and juicy fruit. The best model for estimating firmness contained only the MP for firmness (MPf) as an explanatory variable, whereas the best model for juiciness contained both the MP for juiciness (MPj) and the MPf as explanatory variables. The regression lines from these models suggested that crosses with a low MPf and high MPj can increase the proportion of offspring that have soft fruit and juicy fruit, respectively. However, the slopes of the regression lines for both firmness and juiciness were gentle, indicating that genetic segregation in offspring is wide for both attributes. We infer from these data that softer and juicier fruit can be bred from parents having only moderate ratings for these traits.     

Apoptotic force and tissue dynamics during Drosophila embryogenesis

Understanding cell morphogenesis during metazoan development requires knowledge of how cells and the extracellular matrix produce and respond to forces. We investigated how apoptosis, which remodels tissue by eliminating supernumerary cells, also contributes forces to a tissue (the amnioserosa) that promotes cell-sheet fusion (dorsal closure) in the Drosophila embryo. We showed that expression in the amnioserosa of proteins that suppress or enhance apoptosis slows or speeds dorsal closure, respectively. These changes correlate with the forces produced by the amnioserosa and the rate of seam formation between the cell sheets (zipping), key processes that contribute to closure. This apoptotic force is used by the embryo to drive cell-sheet movements during development, a role not classically attributed to apoptosis.