Reproductive phenotypes in mice with targeted disruption of the 20alpha-hydroxysteroid dehydrogenase gene

In the corpus luteum of rats and mice, 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) catalyzes the conversion of progesterone to a biologically inactive metabolite, 20alpha-dihydroprogesterone (20alpha-OHP). The reduction of progesterone by 20alpha-HSD is believed to be important for functional luteolysis in these rodent species. In addition to the corpus luteum, expression of 20alpha-HSD has been demonstrated in tissues such as the placenta, endometrial epithelia, and fetal skin, although the roles it plays in the latter tissues remain to be determined. To determine the contribution of 20alpha-HSD to functional luteolysis and to the rodent reproductive system more generally, we generated a strain of mice with targeted disruption of the 20alpha-HSD gene. In the 20alpha-HSD-/- mice we obtained, which lacked the genomic region essential for catalytic reaction, neither 20alpha-HSD activity in the corpus luteum nor an increase in the serum concentrations of 20alpha-OHP during pseudopregnancy or pregnancy was detected. The durations of the estrous cycle, pseudopregnancy, and pregnancy were significantly prolonged in the 20alpha-HSD-/- mice, although the serum progesterone levels decreased to levels low enough for delivery of pups at term of pregnancy. In addition, the number of pups, especially live pups, was markedly decreased in the 20alpha-HSD-/- mice. These findings suggest that the role of 20alpha-HSD in functional luteolysis is relatively minor but that it is involved in the survival of newborn mice.     

The product of the human c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity

Antibodies were raised against a synthetic peptide corresponding to 14 amino acid residues at the COOH-terminus of a protein deduced from the human c-erbB-2 nucleotide sequence. These antibodies immunoprecipitated a 185-kilodalton glycoprotein from MKN-7 adenocarcinoma cells. Incubation of the immunoprecipitates with (gamma-32P)ATP resulted in the phosphorylation of this protein on tyrosine residues. These results indicate that the human c-erbB-2 gene product is the 185-kilodalton glycoprotein that is associated with tyrosine kinase activity. Although the c-erbB-2 protein was predicted to encode a protein very similar to epidermal growth factor (EGF) receptor, EGF did not stimulate this kinase activity either in vivo or in vitro.     

Azolla incorporation and dual cropping influences CH4 and N2O emissions from flooded paddy ecosystems

ABSTRACT

To investigate the influence of Azolla (A. filiculoides Lam.) incorporated as a green manure and its subsequent growth as a dual crop with rice on simultaneous methane (CH₄) and nitrous oxide (N₂O) emissions from a flooded alluvial soil planted with rice, a pot experiment with three treatments, chemical fertilizers (NPK) as the control, incorporation of Azolla as green manure (AGM), and AGM plus basal chemical fertilizers (NPK+AGM) was conducted in Tsuruoka, Yamagata, Japan in 2017. AGM and NPK+AGM treatments significantly increased CH₄ emissions at early rice growth stages before 63 days after transplanting (DAT) by 123.0% and 176.7% compared to NPK, respectively. At late rice growth stages (after 63 DAT), only the NPK+AGM treatment significantly increased CH₄ emission by 22.1% compared to NPK. However, percentage of CH₄ emitted after 63 DAT relative to the seasonal CH₄ emission followed the order of NPK (86.2%) > AGM (76.5%) > NPK+AGM (73.3%). Higher CH₄ emissions from AGM and NPK+AGM before 63 DAT were attributed to the incorporated Azolla, while the higher emissions after 63 DAT in all treatment groups were ascribed to rice photosynthesis. AGM and NPK+AGM treatments significantly decreased N₂O emissions by 71.6% and 81.1% compared to NPK, respectively, at early rice growth stages. Azolla incorporation may have restricted N₂O emission from initial soil nitrate before 63 DAT and not have contributed to N₂O emissions after 63 DAT. Significantly higher grain yields were observed under the AGM (32.5%) and NPK+AGM (36.3%) compared to NPK. Together, AGM and NPK+AGM treatments significantly increased seasonal CH₄ emissions by 31.5% and 43.5%, and decreased seasonal N₂O emissions 3.4- and 4.6- fold compared to NPK, respectively. There were no significant differences in the CH₄ emissions per grain yield among the treatments; however compared to NPK, AGM and NPK+AGM treatments significantly reduced N₂O emissions per grain yield by 78.7% and 84.1%, respectively.     

Impact of fresh and aged palm shell biochar on N2O emissions,soil properties,nutrient content and yield of Komatsuna (Brassica rapa var. perviridis) under sandy soil conditions

ABSTRACT

Biochar can reduce N₂O emissions and it can be added to the soil once, whereas fertilizers are often applied every cultivation season. The aging of biochar in soil affects its functioning but it is unclear whether palm shell biochar (PSB) could still mitigate N₂O emissions even when additional basal N fertilizers are applied 1 year after the initial biochar application. We studied the impact of fresh and aged PSB (0%, 6%, 12%, and 18% w/w of dry soil) on N₂O emissions, soil properties, nutrient content and yield of Komatsuna (Brassica rapa var. perviridis) under sandy soil conditions. The aged PSB non-significantly reduced N₂O emissions but significantly offset soil acidification, and maintained a high soil nutrient status. Biochar application with fertilizer significantly increased plant tissue K and Ca content but decreased N, P and Mg content compared to the treatments without biochar. At higher application rates, biochar had negative effects on crop yield but as it aged, the negative effects were offset as a result of the similar variation in plant N uptake. Since seasonal N fertilizer application seems to be inevitable in Komatsuna cultivation, addition of biochar could be a possible way of counteracting the effects of excessive fertilizer use. Further research is needed to assess the feasible biochar application rates for Komatsuna fields in various soil types under field conditions.     

Relationship between increment of groundwater level at the beginning of irrigation period and paddy filed area in the Tedori River Alluvial Fan Area, Japan

There are many paddy fields and large amounts of groundwater in the Tedori River Alluvial Fan in Ishikawa Prefecture, Japan. Water infiltration from paddy fields during irrigation may significantly contribute to groundwater recharge. Groundwater recharge is known to be one outcome of paddy farming, and in general is usually related to land use. However, a decreased area of paddy fields because of socioeconomic factors such as urbanization and increasing area of fallow fields has possibly affected the groundwater environment. Evaluation of the quantitative effect of paddy fields on groundwater is necessary for groundwater conservation. This study examined the relationship between differences in the depth of groundwater from just before the irrigation period to just after the first irrigation of paddy fields (increments of groundwater levels) in observation wells and the area of paddy fields around each well. The paddy areas within circular buffer zones, which were delineated at 0.2 km intervals between 0.2 and 2.0 km centered on each observation well, were calculated. A positive relationship was found between the rise in groundwater and the area of paddy field within different buffer zones at most wells. In addition, in the middle or upper part of the fan, the effect of changes in the area of paddy fields surrounding the well on the groundwater level rise was greater than that on the lower part of the fan.     

Changes in the pericellular connective tissue and breaking strength of the three types of muscles of the cultured carp Cyprinus carpio during storage in ice

To elucidate the structural changes in pink (P), white (W), and red (R) muscles during storage in ice, we measured the breaking strength and changes in pericellular connective tissues of cultured carp. The breaking strength just after killing was highest in R muscle (1.00 ± 0.20 N), lowest in W muscle (0.37 ± 0.07 N), and intermediate (0.84 ± 0.12 N) in P muscle. During the storage period, the breaking strength decreased first in R muscle, then in P muscle, followed by W muscle. The diameter of muscle fibers was greater in W muscle (113 ± 15 μm) than in P muscle (72 ± 3 μm) and R muscle (48 ± 2 μm). Destruction of the honeycomb structure of the pericellular connective tissue occurred most rapidly in W muscle and most slowly in R muscle. These results suggest that the interposing of P muscle fibers in the dorsal ordinary muscle contributes to the acceleration of post-mortem tenderization in fish.     

Differential Expression of Two Cytosolic Ascorbate Peroxidases and Two Superoxide Dismutase Genes in Response to Abiotic Stress in Rice

Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide t...     

Intravenous Administration of Azumolene to Reverse Malignant Hyperthermia in Swine

Background: The efficacy of intravenous (IV) administration of azumolene (Az), an analogue 30‐fold more soluble than dantrolene, on pigs susceptible to malignant hyperthermia (MH) is incompletely understood. Objective: To evaluate efficacy of Az on MH crisis in pigs. Animals: Eight normal (MHN) and 7 susceptible to MH (MHS) pigs (Landrace × Large White × Pietran). Methods: Prospective, laboratory trial. Hypermetabolic crisis was observed in MHS pigs, but not in MHN pigs, after a combined administration of inhaled halothane (1.5%) and IV injection of succinylcholine (SCh; 2.5 mg/kg). Susceptibility was confirmed using a caffeine and halothane contracture test. Az was administered 15 minutes after administration of SCh. Results: Respiratory acidosis (pH 7.16 ± 0.02; Pco ₂, 46.2 ± 9.1 mmHg, HCO₃, 22.5 ± 2.3 mmol/L), fever (38.2 ± 1.1°C), cardiac arrhythmias, and muscle contracture were observed in MHS pigs. MHS pigs (n = 5) treated with Az (2 mg/kg IV) survived the crisis with attenuation of signs (pH 7.30 ± 0.10; Pco ₂, 36.3 ± 4.5 mmHg; HCO₃, 22.9 ± 2.3 mmol/L) and recovery of normal muscle tone and cardiac rhythm. Conclusions and Clinical Importance: Az represents a possible substitute for dantrolene to reverse MH crisis in susceptible pigs.     

CMA staining analysis of chromosomes in several species of Aurantioideae

Fluorochrome staining with chromomycin A₃ (CMA) was used to characterize and compare the CMA banding patterns of chromosomes of 17 species from 13 genera of Aurantioideae, which is one of the seven subfamilies of Rutaceae. All species used in this study had 2n = 18 chromosomes. These chromosomes were classified into five types based on the number and position of CMA-positive bands; B: one telomeric and one proximal band, C: two telomeric bands, D: one telomeric band, E: without bands and Dst: type D with a satellite chromosome. Each species possessed two or four types of chromosomes and unique CMA banding patterns. The CMA banding patterns were 6D + 12E in Clausena anisata (Willd.) Hook. f.,1C + 3D + 14E in Murraya koenigii (L.) Spreng., 1C + 11D + 6E in Murraya paniculata (L.) Jack, 1B + 1C + 10D + 6E in Merrillia caloxylon (Ridl.) Swing., 1C + 9D + 7E + 1Dst in Paramignya lobata Burkill, 2C + 5D + 11E in Severinia buxifolia (Poir.) Tenore, 5D + 13E in Hesperethusa crenulata (Roxb.) Roem., 16D + 2E in Citropsis gabunensis (Engl.) Swing. et M. Kell, 14D + 4E in Citropsis schweinfurthii (Engl.) Swing. et M. Kell, 8D + 10E in Atalantia monophylla DC., 1C + 3D + 14E in Atalantia ceylanica (Arn.) Oliv., 2D + 16E in Atalantia roxburghiana Hook. f., 18E in Swinglea glutinosa (Blanco) Merr., 2D + 16E in Aegle marmelos (L.) Corr., 4D + 14E in Afraegle paniculata (Schum.) Engl., 2B + 2D + 13E + 2Dst in Feronia limonia (L.) Swing. and 3B + 9D + 6E in Feroniella oblata Swing. The advanced genera in each tribe and subtribe such as Merrillia and Feroniella showed heterochromatin (CMA-positive bands) rich chromosome configuration in general. Swinglea glutinosa without heterochromatin rich chromosome, which is considered to be the most primitive chromosome configuration, seems to be the key genus for clarifying the evolution of the tribe Citreae.     

Effect of temperature decrease on oocyte development,sex steroids,and gonadotropin β-subunit mRNA expression levels in female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To improve understanding of the mechanism of early ovarian development in eels, the effects of water temperature decrease on oocyte development, plasma levels of sex steroids [estradiol 17β (E2), testosterone (T), 11-ketotestosterone (11-KT)], and gonadotropin β-subunit [follicle-stimulating hormone (FSHβ), luteinizing hormone (LHβ)] messenger RNA (mRNA) expression levels were investigated. A total of 27 female Japanese eels Anguilla japonica were divided into initial, control, and test (water temperature decrease) groups. Starting on 22 September 2009, eels in the test group were reared in a tank with gradual temperature decrease from 25°C to 15°C over 39 days, while the control group was maintained at 25°C. The test group accumulated more oil droplets in their oocytes than did the other groups. Levels of sex steroids, especially 11-KT, were higher in the test group. In contrast, FSHβ and LHβ mRNA expression levels were lower in the test group. These results suggest that water temperature decrease only induced an early stage of ovarian development that was partly affected by an 11-KT increase. For further maturation, other environmental factors related to induction of gonadotropin increase appear to be needed.