Correlations of the electrophysiological activity of neonicotinoids with their binding and insecticidal activities

The electrophysiological actions of various neonicotinoids, including substituted benzyl derivatives, against recombinant Drosophila SAD/chicken beta2 hybrid nicotinic acetylcholine receptor (nAChR) were measured to analyze the relationships between the in vivo (insecticidal) and in vitro (binding and agonist) activities. Most of the neonicotinoids tested were capable of inducing inward currents by activating the hybrid nAChRs expressed in Xenopus laevis oocytes, whereas some compounds had no agonist activity and only blocked the acetylcholine-induced currents. Variations in the agonist activity were well correlated with those in the binding potency evaluated using [3H]imidacloprid as well as insecticidal activities.     

Genetic variation in wood stiffness and strength properties of hybrid larch (Larix gmelinii var. japonica × L. kaempferi)

Genetic parameters for wood stiffness and strength properties were estimated in a 29-year-old hybrid larch stand (Larix gmelinii var. japonica × Larix kaempferi). The study included 19 full-sib larch families from Hokkaido, northern Japan. Implications of these genetic parameters in wood quality improvement are subsequently discussed. Traits included in the analyses were the dynamic modulus of elasticity of green logs (E _log), the modulus of elasticity (MOE), the modulus of rupture (MOR), compression strength parallel to the grain (CS) in small clear specimens, wood density (DEN), and diameter at breast height (DBH). DEN had the lowest coefficients of variation and MOE the highest. The narrow-sense heritability estimates of E _log, MOE, MOR, and CS were 0.61, 0.44, 0.60, and 0.43, respectively, and those of DEN and all mechanical properties increased from an inner to outer position within the stem. E _log and DEN had high positive phenotypic (0.52–0.83) and genetic (0.70–0.92) correlations with MOE, MOR, and CS. The mechanical properties of the inner position of the stem had rather high phenotypic and genetic correlations with those of the outer position and overall mean. The predicted gains in wood stiffness (E _log and MOE) were higher than those of the strength properties (MOR and CS). The predicted correlated responses in MOE, MOR, and CS when selecting for E _log and DEN were 72.6%–97.8% of a gain achievable from direct selection of these traits. DBH showed an insignificant correlation with all mechanical properties, although selection of this trait had a slightly negative effect on the mechanical properties.     

Production of transgenic cloned pigs expressing the far-red fluorescent protein monomeric Plum

Monomeric Plum (Plum), a far-red fluorescent protein with photostability and photopermeability, is potentially suitable for in vivo imaging and detection of fluorescence in body tissues. The aim of this study was to generate transgenic cloned pigs exhibiting systemic expression of Plum using somatic cell nuclear transfer (SCNT) technology. Nuclear donor cells for SCNT were obtained by introducing a Plum-expression vector driven by a combination of the cytomegalovirus early enhancer and chicken beta-actin promoter into porcine fetal fibroblasts (PFFs). The cleavage and blastocyst formation rates of reconstructed SCNT embryos were 81.0% (34/42) and 78.6% (33/42), respectively. At 36–37 days of gestation, three fetuses systemically expressing Plum were obtained from one recipient to which 103 SCNT embryos were transferred (3/103, 2.9%). For generation of offspring expressing Plum, rejuvenated PFFs were established from one cloned fetus and used as nuclear donor cells. Four cloned offspring and one stillborn cloned offspring were produced from one recipient to which 117 SCNT embryos were transferred (5/117, 4.3%). All offspring exhibited high levels of Plum fluorescence in blood cells, such as lymphocytes, monocytes and granulocytes. In addition, the skin, heart, kidney, pancreas, liver and spleen also exhibited Plum expression. These observations demonstrated that transfer of the Plum gene did not interfere with the development of porcine SCNT embryos and resulted in the successful generation of transgenic cloned pigs that systemically expressed Plum. This is the first report of the generation and characterization of transgenic cloned pigs expressing the far-red fluorescent protein Plum.     

Microinsemination with first-wave round spermatids from immature male mice

In several mammalian species, including mice, round spermatids have been used to produce normal offspring by means of microinsemination techniques. In this study, we examined whether mouse round spermatids retrieved from immature testes undergoing the first wave of spermatogenesis had acquired fertilizing ability comparable to cells from mature adults. Microinsemination with round spermatids was performed by direct injection into preactivated oocytes, as previously reported. About 60-85% of the successfully injected oocytes developed to the morula/blastocyst stage after 72 h in culture, irrespective of the age of the males (17-25 days old). After embryo transfer, normal pups were obtained from all age groups, including the day-17 group, the stage at which the first round spermatids appeared. A high correlation (r=0.90) was found between the birth rate and male age (P<0.01, Spearman rank correlation), indicating that the efficiency of producing offspring was dependent on the age of the donor males. Imprinted genes (H19, Igf2, Meg3, and Igf2r) were expressed from the correct parental alleles (maternal, paternal, maternal, and maternal, respectively) in all (n=12) day-9.5 fetuses derived from day-20 spermatids. These results clearly indicate that at least some first-wave spermatogenic cells have a normal haploid genome with the correct paternal imprint and are capable of supporting full-term embryo development, as do mature spermatozoa from adults. The use of male germ cells from immature animals may save time in the production of inbred/congenic strains and rescue male-factor infertility of early onset.     

An experimental study of apico-aortic valved conduit (AAVC) for surgical treatment of aortic stenosis in dogs

A new valved conduit was developed using a canine aortic valve. The bioprosthetic valve was fixed with glutaraldehyde and epoxy compound (Denacol-EX313/810). A vascular graft composed of ultra-fine polyester fiber (10 mm in diameter, 200 mm in length) was used. Four dogs underwent apico-aortic valved conduit (AAVC) implantation and aortic banding (bypass group, BG), while another 4 dogs underwent aortic banding without AAVC implantation (control group, CG). Cardiac catheterization and angiocardiography were performed for assessment of hemodynamics 2 weeks and 6 months after surgery. Left ventricular systolic pressure, left ventricular end-diastolic pressure and the left ventricular-aortic pressure gradient differed significantly (P<0.01) between the BG and CG dogs. Left ventricular angiocardiography showed patency of the valved conduit in all the BG dogs. Echocardiography was performed before and 2, 4 and 6 months after surgery, and showed that while pressure overload caused concentric myocardial hypertrophy in the CG dogs, the left ventricle dilated eccentrically in the BG dogs. Furthermore, relief of left ventricular pressure overload by AAVC was maintained.     

Percutaneous transcatheter coil embolization of a ventricular septal defect in a dog

A 4-month-old male French Bulldog weighing 5.0 kg (11 lb) was referred for a heart murmur. A grade 3/6 systolic murmur was detected at the left heart base and a grade 4/6 systolic murmur was detected at the right heart base. By use of color-flow Doppler ultrasonography and cardiac catheterization, a diagnosis of supracristal ventricular septal defect (VSD) with accompanying aortic regurgitation was made. Percutaneous transcatheter coil embolization was used to close the VSD. Because residual shunt was detected via echocardiography after coil implantation, the residual shunt was followed periodically via echocardiography to detect spontaneous closure of the VSD. Volume overload in the left ventricle was detected in the dog 131 days after admission. Additional coils were placed 137 days after admission. Hemolysis resulting in hemoglobinuria was detected, but this complication was mild. In the dog of this report, results of coil occlusion for correction of VSD were promising. Thus, coil occlusion should be considered as an alternative treatment for VSD in dogs.     

Structural determination of the oligosaccharides in the milk of a giant anteater (Myrmecophaga tridatyla)

There appears to be strong evidence supporting four major clades of placental mammals: (i) Afrotheria; (ii) Xenarthea; (iii) Euarchontoglires; and (iv) Laurasiathera. This is the first study in which the carbohydrates have been analyzed in the milks of Xenarthea. Our aim was to clarify whether the saccharides of giant anteater milk resemble those of other eutherian species, especially of the Asian elephant, which is one of the Afrotherea. We found that lactose is a dominant saccharide in this milk, as it is in that of most eutherians, and that the milk contains smaller amounts of oligosaccharides, whose structures were determined as Gal(α1‐3)Gal(β1‐4)Glc (isoglobotriose), Neu5Ac(α2‐6)Gal(β1‐4)Glc (6'‐N‐acetylneuraminyllactose), Neu5Gc(α2‐6)Gal(β1‐4)Glc (6'‐N‐glycolylneuraminyllactose), Neu5Ac(α2‐6)Gal(β1‐4)GlcNAc(β1‐3)Gal(β1‐4)Glc (LST c) and Neu5Gc(α2‐6)Gal(β1‐4)GlcNAc(β1‐3)Gal(β1‐4)Glc. This pattern of milk carbohydrates is rather close to that of the milks of other eutherian species. It is notable that the milk contained oligosaccharides with Neu5Gc residues but no oligosaccharides containing α(2‐3) linked Neu5Ac residues such as Neu5Ac(α2‐3)Gal(β1‐4)Glc.     

Effects of high potassium chloride supplementation on growth rate and renal function in mice

Fifty‐four mice were assigned to a control diet group or a KCl diet group to clarify the effects of KCl supplementation on growth rate, water intake and renal function in mice, and 5% of KCl was supplemented in KCl diets for 1–4 weeks. Bodyweights of KCl supplemented mice were significantly lower than those of control mice, and bodyweights of control and KCl supplemented mice at 28 days after treatment were 45.5 and 41.2 g, respectively. Feed intake was not affected by KCl supplementation, but water intake of KCl supplemented mice was significantly higher than that of control mice. Bone weights of KCl supplemented mice at 2 weeks after treatment were significantly lower than those of control mice. Serum urea nitrogen concentration at 4 weeks and serum K and Cl concentrations at 2 weeks were significantly lower in KCl supplemented mice. Histological alteration using hematoxylin–eosin and Sirius red staining was not found in the kidney of each mouse at 1, 2, and 4 weeks after treatment. These results suggest that high KCl supplementation decreases the rate of bodyweight gain and increases water intake in mice.     

Coumestrol decreases intestinal alkaline phosphatase activity in post-delivery mice but does not affect vitamin D receptor and calcium channels in post-delivery and neonatal mice

In this study, we investigated the effects of administration of coumestrol during pregnancy on calcium (Ca) metabolism in post-delivery maternal and neonatal mice. From 6.5 to 16.5 days post coitus (dpc), pregnant females were administered daily doses of coumestrol (200 microg/kg body weight/day). One day after parturition, blood samples and the kidneys, liver, jejunum and duodenum were obtained from each of maternal mouse, and blood samples and the kidneys and liver were obtained from neonatal mice. Coumestrol did not have any significant effect on the Ca and inorganic phosphorus concentrations in the sera of the maternal and neonatal mice. No notable effects of coumestrol were observed in relation to Vitamin D receptor expression in the maternal and neonatal mice by immunohistochemical analysis. Coumestrol did not affect the Vitamin D receptor and epithelial calcium channel and 2 mRNA levels in any of the organs investigated. Enzyme histochemical analysis showed that coumestrol decreased intestinal alkaline phosphatase activity in the maternal jejunum and duodenum. In the duodenum, coumestrol decreased expression of intestinal alkaline phosphatase, c-fos and vascular endothelial growth factor at the mRNA level. However, we did not observe any significant effects of coumestrol on the expression of these genes. In conclusion, coumestrol decreased intestinal alkaline phosphatase activity in the small intestines of maternal mice at the level used in the present study, and the mechanisms underlying this effect are different for the jejunum and duodenum.