Effect of Cetrorelix administration on ovarian stimulation in aged mice

In mice, ovarian stimulation via hormone administration is an effective method for obtaining many ova simultaneously, but its effect is reduced by the influence of aging. In this study, we demonstrate that this problem can be improved by administering the gonadotropin-releasing hormone antagonist Cetrorelix prior to ovarian stimulation. Before 12-month-old female mice were injected with 5 IU pregnant mare serum gonadotropin and 5 IU human chorionic gonadotropin, we administered 5 µg/kg Cetrorelix for 7 consecutive days (7 times) or 3 times once every 3 days. As a result, 8.7 ± 1.9 (mean ± SEM, n=10) and 9.8 ± 1.3 (n=10) oocytes were obtained, respectively, as opposed to 4.7 ± 1.2 oocytes (n=9) in the case of no administration. Collagen staining of ovarian tissue showed that Cetrorelix administration reduced the degree of fibrosis, which improved ovarian function. In addition, equivalent fertilization and fetal development rates between control and Cetrorelix-treated aged mouse-derived oocytes were confirmed by in vitro fertilization and embryo transfer (Fertilization rate; control: 92.2% vs. 3 times: 96.9%/7 times: 88.5%, Birth rate; control: 56.4% vs. 3 times: 58.3%/7 times: 51.8%), indicating the normality of the obtained oocytes. It is concluded that Cetrorelix improved the effect of superovulation in aged mice without reducing oocyte quality. This procedure will contribute to animal welfare by extending the effective utilization of aged female breeding mice.     

Differential impact of liana colonization on the leaf functional traits of co-occurring deciduous and evergreen trees in a tropical dry scrub forest

The present study was carried out to analyze the leaf functional traits of co-occurring evergreen and deciduous tree species in a tropical dry scrub forest. This study also intended to check whether the species with contrasting leaf habits differ in their leaf trait plasticity, responding to the canopy infestation by lianas. A total of 11 leaf functional traits were studied for eight tree species with contrasting leaf habits (evergreen and deciduous) and liana-colonization status (with or without liana). In the liana-free environment (L^–), evergreen trees had significantly higher leaf tissue density (LTD) and total chlorophyll (CHL_t) than the deciduous species. Whereas the deciduous trees had higher specific leaf area (SLA) and mass-based leaf nitrogen concentration (N_mass). The leaf trait-pair relationship in the present study agreed with the well-established global trait-pair relationships (leaf thickness (LT) vs. SLA, N_mass vs. LT, SLA vs. N_mass, and LDMC vs. SLA). There was a significant difference between L⁺ and L^– individuals in leaf area (LA), petiole length (PL), SLA, LDMC, and CHL_t in the deciduous species. On the other hand, evergreen species showed marked differences across LT, SLA, LTD, N_mass, and chlorophyll components between L⁺ and L^– individuals of the same species. The results revealed the differential impact of liana colonization on the host trees with contrasting leaf habits. The deciduous species with the acquisitive strategy can have a competitive advantage over evergreen species in the exposed environments (L^–), whereas evergreen species with shade-tolerant properties were better acclimated to the shaded environments (L⁺). Therefore, liana colonization can significantly impact the C-fixation strategies of the host trees by altering their light environment and further, the magnitude of such impact may vary among species of different leaf habits. The result also indicated the patterns of convergence and divergence in some of the leaf functional traits between evergreen and deciduous species explaining the patterns of species co-existence.

     

D-dihexanoylphosphatidylcholine is not a pure competitive inhibitor of phospholipase A2 hydrolysis of L-dihexanoylphosphatidylcholine

The inhibition of Crotalus adamanteus phospholipase A2 hydrolysis of L-dihexanoylphosphatidylcholine by D-dihexanoylphosphatidylcholine was investigated with inhibitor and substrate in the monomeric concentration range. The results showed that the D-enantiomer acts as a partial (not pure) competitive inhibitor of the enzyme. These results suggest that an ESI complex exists, in which hydrolysis of the substrate still occurs. Thus, binding of the D-enantiomer to the enzyme decreases the affinity for the substrate by a factor, alpha, while Vmax is unaffected. The value of alpha was determined to be 4.70 +/- 0.14. These findings complicate the use of D-phosphatidylcholines in mixed micelles with the L-enantiomer as a possible method to investigate the mechanism of interfacial activation of this enzyme.     

Contamination and health risk assessment of trace elements in soil at play centers of urban low-income settings

Abstract

Young children are considered critical receptors of potentially toxic trace elements (PTEs) by non-dietary ingestion of contaminated soil. The study assessed the potential enrichment of soil and the health risk of PTEs to 471 children less than seven years via non-dietary soil ingestion at six Early Childhood Development Centers (ECDCs) in urban low-income settings. The total concentrations of PTEs were determined by ICP-AES after wet acid digestion. The extent of soil contamination with PTEs and their source apportionment were assessed by the enrichment factor (EF). The US-EPA risk assessment model was used to determine the risk of PTE exposure by children. Multivariate statistical analyses and the EF suggested anthropogenic origin of PTEs in playgrounds and indoors, especially Cd and Pb from atmospheric deposition. Indoor floor dust at ECDCs was enriched (significant to extreme) with PTEs of anthropogenic origin imported from the outside environment. Children at the six ECDCs were not at significant non-carcinogenic risk of PTEs in soil and dust through non-dietary ingestion. The study setting is typical of urban child play centers in low-income countries which needs regular risk assessment and the enforcement of legislation in order to reduce the exposure of children to PTEs.     

Conservation genetics of bull trout: Geographic distribution of variation at microsatellite loci

We describe the genetic population structure of65 bull trout (Salvelinus confluentus)populations from the northwestern United Statesusing four microsatellite loci. Thedistribution of genetic variation as measuredby microsatellites is consistent with previousallozyme and mitochondrial DNA analysis. Thereis relatively little genetic variation withinpopulations (H ^S = 0.000 – 0.404,average H ^S = 0.186, but substantialdivergence between populations (F ^ST = 0.659). In addition, those populations that had low genetic variation forallozymes also tended to have low geneticvariation at microsatellite loci. Microsatellite analysis supports the existenceof at least three major geneticallydifferentiated groups of bull trout: (1)``Coastal' bull trout populations, (2) ``SnakeRiver' populations, which also include the JohnDay, Umatilla, and Walla Walla Rivers and, (3)populations from the upper Columbia River,primarily from the Clark Fork basin. Withinthe major assemblages, populations are furthersubdivided, primarily at the river basin level. Most of the genetic similarities we havedetected probably reflect patterns of historicisolation and gene flow. However, in somecases, genetic drift and low levels ofvariation appear to have influenced therelationships inferred from these data. Finally, we suggest using a hierarchicalapproach to direct management actions inspecies such as bull trout for which most ofthe genetic variation exists among populationsand local populations in close proximitytypically are genetically distinct.     

Purification and properties of multiple forms of mouse trypsinogen

Three trypsinogens and one chymotrypsinogen were found in and purified from the pancreas of a mouse strain (CFO). The molecular weights of the trypsinogens and the chymotrypsinogen were all estimated as 25 000. The enzyme properties of the three trypsinogens were studied and they showed very low K_m values (3.2?6.5 μM) for the substrates, BzArgOEt and TosArgOMe, and the dame pH optimum profile between pH 8.0–10.0. However, the ratios of catalytic rate constants, k_cat (s^?1), with BzArgOEt as substrates compared to that with TosArgOMe were very different. The values of Try-III were similar with the two substrates, Try-I was slightly higher value with TosArgOMe than with BzArgOEt, and the values of Try-II were much higher with TosArgOMe than with BzArgOEt. Also, the trypsinogens and the chymotrypsinogen were purified from pancreas of Mol-A strain mice. When the enzyme properties of the three trypsinogens were examined, one form of trypsinogen (Try-I) was shown to have different properties in k_cat (s^?1) for the two substrates, compared to the trypsinogen of CFO mice.     

Identification of the facultative heterochromatic X chromosome in females of 25 rodent species.

Treatment of the chromosomes of 25 rodent species with a 50 degrees C hypotonic solution and Giemsa staining permitted identification of the heterochromatic X chromosome in 24 species. With this technique, the facultative of the heterochromatic X chromosome or the facultative portion of large, composite-type X chromosoms is stained darker than the other chromosomes, allowing it to be distinguished from the homologous euchromatic X chromosome in female metaphase cells. Intense staining of the single X chromosome was not observed in male metaphase cells. It is suggested that this differential staining of one of the two X chromosomes might be due to qualitative differences in chromosomal proteins rather than to differences in the degree of chromosomal condensation or in DNA base sequence.     

Amino Acid Nucleic Acids: Synthesis and Hybridization Properties of a Novel Class of Antisense Oligonucleotides

Abstract

Oligonucleotides containing novel phoshoramidite 12 were synthesized and studied for their hybridization properties for the first time. Interestingly, these modified oligonucleotides showed a remarkable resistance to exonuclease.