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.     

Seasonal variations in major components of Crassostrea gigas from Seto Inland Sea

Fisheries Science - The proximate composition and nutritional content of oysters define their quality and commercial value. Cultivation site and harvest date may impact these characteristics. We...     

Changes in the charged metabolite and sugar profiles of pasteurized and unpasteurized Japanese sake with storage

Japanese sake (rice wine) is commonly heat treated (pasteurized) to maintain its quality. In this study, temporal changes in the metabolite profiles of pasteurized and unpasteurized sake were investigated during storage. Metabolomic analyses were conducted for eight sets of pasteurized and unpasteurized sake obtained from single process batches stored at 8 or 20 °C for 0, 1, 2, or 4 months. Capillary electrophoresis time-of-flight mass spectrometry and liquid chromatography tandem mass spectrometry were used to obtain charged metabolite and sugar profiles, respectively. The total amino acid concentration decreased with storage, and the decrease was faster in pasteurized sake than in unpasteurized. The organic acid concentrations were relatively constant in both types of sake. Peptide and glucose concentrations increased and polysaccharide concentrations decreased in unpasteurized sake, while they were relatively constant in pasteurized sake. Rather than stabilizing the sake metabolite profile during storage, pasteurization results in characteristic changes compared to unpasteurized sake.     

Ultrasonographic patterns,clinical findings,and prognostic variables in dogs from Asia with gallbladder mucocele

BackgroundGallbladder mucocele (GBM) is a common biliary disorder in dogs that can be categorized into 6 types, but the value of this classification scheme remains unknown. Cholecystectomy is associated with high death rates and warrants additional interrogation.ObjectivesInvestigate the clinical value of ultrasonographic diagnosis of type of GBM and identify prognostic factors in dogs with GBM undergoing cholecystectomy.AnimalsTwo hundred sixteen dogs.MethodsRetrospective cohort study. Dogs with GBM diagnosed from 2014 to 2019 at 6 veterinary referral hospitals in Asia. Ultrasonogram images were reviewed and a GBM type (ie, types I‐VI) assigned.ResultsDogs with GBM type V as compared to I (OR, 8.6; 95% CI, 2.6‐27.8; P < .001) and III (OR, 10.0; 95% CI, 2.5‐40.8; P = .001), and dogs with type VI compared to I (OR, 10.5; 95% CI, 1.8‐61.2; P = .009) and III (OR, 12.3; 95% CI, 1.8‐83.9; P = .01) were more likely to exhibit signs of biliary tract disease. Independent predictors of death after cholecystectomy included age (OR, 2.81; 95% CI, 1.41‐5.59; P = .003) and intraoperative systolic blood pressure (SBP) nadir. There was an interaction between SBP nadir and gallbladder rupture; SBP nadir in dogs with (OR, 0.92; 95% CI, 0.89‐0.94; P < .001) and without (OR, 0.88; 95% CI, 0.82‐0.93; P < .001) gallbladder rupture.Conclusion and Clinical ImportanceIncreasing developmental stage of GBM could be associated with an increased likelihood of biliary tract related clinical signs. Nadir SBP deserves further investigation as a prognostic or potentially modifiable variable, particularly in the presence of gallbladder rupture.     

Early establishment of broadleaved trees after logging of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> and <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> plantations with different understory treatments

We studied the effects of the amount of advanced regeneration before logging and different understory treatments on forest recovery, particularly the establishment of lucidophyllous (evergreen broadleaved) species, following the logging of conifer plantation with a focus on the sprouting ability. The establishment of lucidophyllous tree and shrub species originating from advanced regeneration was closely related to the total amount of these elements in the advanced regeneration, regardless of understory treatment. The understory-left treatment could preserve understory as surviving individuals by high proportion (60% or more of the understory) compared with the understory-cleared practice after logging. However, half of the surviving individuals were damaged to some extent by logging and skidding. Relatively large trees were likely to survive with no damage, indicating that they face a lower risk than smaller trees. In understory-cleared stands, a high proportion (60% or more) of the understory regenerated by resprouting. However, the resprouting ratio depended on tree size and species. Therefore, as well as the amount of the understory, the tree size of understory before logging would be also an important factor for early forest recovery. Further, the infrequent (low-density) species could easily disappear after logging because of their low resprouting ratio and low density. Thus, recovery from resprouting alone would risk simplification of the species composition after logging. We concluded that leaving the developed understory intact before logging can enhance the speed and biodiversity of forest recovery.     

Reproductive characteristics of Japanese monkeys on Shimokita Peninsula,Japan, the northernmost habitat of wild primates in the world

The Shimokita Peninsula in Aomori Prefecture, Japan, which is inhabited by Japanese monkeys (Macaca fuscata), is the northernmost habitat for wild primates in the world. This study was the first to determine the conception dates of specific individuals and estimate the pregnancy rate of wild populations in this region. The pregnancy rate of animals aged 5 years or more at delivery was estimated to be 40.9% (27/66). Conception dates of each fetus were also estimated using a regression line of Pig-tail monkeys (Macaca nemestrina), which are taxonomically related to Japanese monkeys and have a similar physique. The conception dates were distributed across 90 days between September 24th and December 23rd, with a mean conception date of November 4th (SD=22.3 days, n=53). Using these findings, the mean birth date was estimated as April 25th, more than two weeks earlier than the mean birth date in previous research determined using direct observations carried out over the past 20 years ago. Global warming due to climate change is thought to be one of the main causes of this difference.     

Motility and fertility of cryopreserved spermatozoa of the Japanese sea cucumber Apostichopus japonicus

We developed both a cryopreservation method for Japanese sea cucumber spermatozoa and an artificial fertilization method using post‐thaw spermatozoa. Twenty per cent dimethyl sulfoxide (DMSO), 16% foetal bovine serum, and 64% artificial seawater were suitable cryodiluent, and the diluent was pre‐cooled to 0°C. Semen was diluted with the solution and enclosed in a 250 μl straw, cooled to ?50°C at 10.4 ± 0.4°C/min, and immediately immersed in liquid nitrogen. Although this method showed the highest post‐thaw motility in all the conditions we examined, its post‐thaw motility was still less than approximately 15%. Artificial fertilization was carried out by adding post‐thaw semen with a cryodiluent to the oocytes. The fertilization rate of 200 oocytes/ml seawater increased with the amount of post‐thaw semen from 1 to 5 μl but showed a significant decrease at 25 μl. This decrease was considered to be due to DMSO in the cryodiluent, because the fertilization rate of the fresh semen decreased sharply when the DMSO concentration around the oocytes was 1.0% or more. Further improvement in increasing post‐thaw motility and lowering the cryoprotectant concentration is necessary for commercial‐scale artificial fertilization.     

Synthesis and insect antifeedant activity of aurones against Spodoptera litura larvae

A series of aurones were prepared from various phenols via phenoxy acetic acids and coumaranones and evaluated for insect antifeedant activity against the common cutworm (Spodoptera litura). The naturally occurring aurone was most active at an ED50 of 0.12 micromol/cm2. The synthetic precursor, coumaranones, showed that the introduction of methoxyl and methyl groups to the benzene ring increased insect antifeedant activity. Similarly, the tested aurones showed that the introduction of methoxyl group to the A and/or B rings increased the insect antifeedant activity, but 4,5,6- and 3',4',5'-trisubstituted compounds did not show this activity in this test. The hydroxylation of aurones in the B ring should be disadvantageous for insect antifeedant activity against S. litura. Although the melting points did not correlate well with the insect antifeedant activity, compounds that were nearly inactive had high melting points. A significant correlation was noted between biological activity (pED50) and a hydrogen-bonding parameter calculated from the Rf value obtained from SiOH thin-layer chromatography and a lipophilicity parameter (log k) calculated from the retention time in ODS high-performance liquid chromatography. The respective correlation coefficients (r) were -0.83 and -0.70. The introduction of alkoxy and alkyl groups along with adequate hydrogen bonding seems to contribute to the antifeedant activity of the compounds tested.     

Improvement of cumulus-free oocyte maturation in vitro and its application to microinsemination with primary spermatocytes in mice

In micromanipulation experiments using immature oocytes, final ooplasmic maturation is often compromised because the oocytes are usually first freed from their nurturing cumulus cells. This study was undertaken to determine whether cumulus-free in vitro maturation (IVM) in mice could be improved by modifying IVM medium having defined components. Cumulus-free germinal vesicle (GV) stage oocytes were subjected to IVM in either alphaMEM medium, TYH medium, or a 1:1 mixture of the two (termed TaM). TYH medium produced a better maturation rate (181/196; 92.3%) than alphaMEM (184/257; 71.6%). However, alphaMEM supported better embryo development to the morula/blastocyst stage than TYH following in vitro fertilization (93.3% vs. 76.5%) or parthenogenetic activation (82.4% vs. 60.4%). Mitochondrial distribution in MII oocytes was diffuse following IVM in alphaMEM, but was aggregated with TYH. The maturation promoting factor (MPF) activity in MII oocytes was significantly higher in TYH than in alphaMEM (P<0.05). Oocytes cultured in TaM had intermediate characteristics and essentially resembled in vivo matured oocytes, with the mitochondrial distribution pattern being most typical of that condition. The highest rate of development from GV oocytes to full-term fetuses following in vitro fertilization and embryo transfer to foster mothers (23.8%) was obtained using TaM. When this IVM system was applied to MI oocytes injected with spermatocytes, offspring were first obtained without cytoplasmic replacement at MII. Thus, optimization of the culture medium can considerably improve the quality of cumulus-free oocyte IVM in mice.