The simultaneous analysis of mixed discrete and continuous outcomes using nonlinear threshold models

Mixed discrete and continuous outcomes are commonly measured on each experimental unit in dose-response studies in toxicology. The dose-response relationships for these outcomes often have dose thresholds and nonlinear patterns. In addition, the endpoints are typically correlated, and a statistical analysis that incorporates the association may result in improved precision. We propose an extension of the generalized estimating equation (GEE) methodology to simultaneously analyze binary, count, and continuous outcomes with nonlinear threshold models that incorporates the intra-subject correlation. The methodology uses a quasi-likelihood framework and a working correlation matrix, and is appropriate when the marginal expectation of each outcome is of primary interest and the correlation between endpoints is a nuisance parameter. Because the derivatives of threshold models are not continuous at each point of the parameter space, we describe the necessary modifications that result in asymptotically normal and consistent estimators. Using dose-response data from a neurotoxicity experiment, the methodology is illustrated by analyzing five outcomes of mixed type with nonlinear threshold models. In this example, the incorporation of the intra-subject correlation resulted in decreased standard errors for the threshold parameters.     

Two new chalcone glycosides from the stems of Entada phaseoloides

Two new chalcone glycosides 4′-O-(6″-O-galloyl-β-d-glucopyranosyl)-2′,4-dihydroxychalcone (1) and 4′-O-(6″-O-galloyl-β-d-glucopyranosyl)-2′-hydroxy-4-methoxychalcone (2) together with one known chalcone glycoside 4′-O-β-d-glucopyranosyl-2′-hydroxy-4-methoxychalcone (3) were isolated from the stems of Entada phaseoloides. The structures of the new compounds were elucidated on the basis of extensive spectroscopic analysis, including HSQC, HMBC, ¹H–¹H COSY, and chemical evidences. This is the first report of chalcone-type compounds isolated from the genus Entada.     

Regulation of mammalian tooth cusp patterning by ectodin

Mammalian tooth crowns have precise functional requirements but cannot be substantially remodeled after eruption. In developing teeth, epithelial signaling centers, the enamel knots, form at future cusp positions and are the first signs of cusp patterns that distinguish species. We report that ectodin, a secreted bone morphogenetic protein (BMP) inhibitor, is expressed as a "negative" image of mouse enamel knots. Furthermore, we show that ectodin-deficient mice have enlarged enamel knots, highly altered cusp patterns, and extra teeth. Unlike in normal teeth, excess BMP accelerates patterning in ectodin-deficient teeth. We propose that ectodin is critical for robust spatial delineation of enamel knots and cusps.     

成团泛菌低分子脂多糖的急性毒性、刺激性、致敏性及遗传毒性评估

本研究对成团泛菌低分子脂多糖（Pantoea agglomerans lipopolysaccharide,LPSp）的安全性进行初步评估。本研究采用一次限量法,用昆明种小鼠进行LPSp急性经口毒性试验,了解LPSp的急性毒性;采用新西兰兔分别进行LPSp急性和多次皮肤刺激性试验以及急性眼刺激性试验,了解LPSp的皮肤和粘膜刺激性;采用豚鼠进行LPSp皮肤变态反应试验,了解LPSp的致敏性;应用平板掺入法进行鼠伤寒沙门氏菌/回复突变试验和小鼠骨髓细胞微核试验考察LPSp的遗传危害。急性毒性试验结果显示,LPSp对小鼠经口一次灌胃的LD50大于5000mg/kg体重,属实际无毒级别;LPSp急性和多次皮肤刺激性试验以及急性眼刺激性试验结果显示,皮肤刺激和眼刺激积分均为0分,LPSp对皮肤无刺激性、对眼睛无急性刺激性;在皮肤变态反应试验中,LPSp在各观察时间点的皮肤变态反应积分均为0分,其致敏率均为0%,说明LPSp对豚鼠无致敏性;LPSp的鼠伤寒沙门氏菌/回复突变试验结果呈阴性（P〉0.05）;LPSp的小鼠骨髓细胞微核试验结果亦呈阴性,LPSp各剂量组的微核发生率与阴性对照组未见统计学差异（P〉0.05）,而与阳性对照组有明显差异（P〈0.01）。本研究结果表明,在本实验剂量范围内,LPSp对小鼠经口毒性极低,属实际无毒级别,对家兔皮肤和眼睛无明显刺激性,对豚鼠无致敏性,对所试菌株和小鼠体细胞无诱变性和致突变性。     

Mouse embryo co‐culture with autologous cumulus cells and fetal development post‐embryo transfer

This study was conducted to examine the potential for implantation and sustainable fetal development of mouse embryos cultured from the pronuclear to blastocyst stage. Pronuclear embryos from ICR mice (Harlan Sprague‐Dawley) were cultured in Sydney IVF sequential media (Cook) to the blastocyst stage in medium only or co‐cultured with autologous cumulus cells. We also experimented with co‐culture in 100 µL drops. Drop co‐culture produced blastocyst formation rates with a mean of 47.0%, which was significantly higher (P < 0.05) compared to embryos cultured in identical culture conditions except without cumulus cells at 27.3%. Blastocysts obtained in vitro in Cook medium only and co‐cultured in Cook medium with cumulus cells were transferred to pseudopregnant females of ICR strain. The day of blastocyst transfer into surrogate females was designated as post‐transfer of blastocyst day 1 (PT 1). The implantation and fetal development was compared to embryo transfer of in vivo derived blastocysts, which served as controls. There were no statistical differences for implantation and fetal development rates for blastocysts cultured in vitro in either Cook medium only or co‐culture in Cook medium with cumulus cells compared to in vivo‐derived blastocysts. The advantage of the co‐culture system is in generating more blastocysts available for transfer.     

Characteristics of fertilizer uptake and biodistribution in strawberry plants in two Japanese cultivars in hydroponic culture

ABSTRACT

The fertilizer absorption characteristics of strawberries are not clear, although appropriate fertilization is definitely necessary to ensure produce quality and quantity. This study aimed to determine the amounts of macro- and micronutrients absorbed during cultivation of strawberries and their biodistribution and utilization in the plant body. We cultivated Japanese strawberries ‘Benihoppe’ and ‘Kirapika’ in small hydroponic equipment containing a nutrient solution and determined the amounts of N, P, K, Ca, Mg, Fe, Mn, B, Zn, Cu, and Mo absorbed during and at the end of cultivation. The results revealed the adsorption levels of these elements during the cultivation period. The nutrient concentrations varied greatly among plant organs. In particular, P and B accumulated at high levels in the leaves and stem, K, Ca, Mg, Mn, Zn, and Cu accumulated in the crown, and N, Fe, and Mo accumulated in the roots. In addition, the uptake levels of N, P, K, Mg, Mn, Zn, and Cu differed between Benihoppe and Kirapika. Our results provide useful information for determining fertilizer application rates in strawberry cultivation.     

Food-drug interaction between ferulic acid and nateglinide involving the fluorescein/H+ cotransport system

In clinical, patients usually take many kinds of drugs at the same time. Thus, drug-drug interactions involving transporters can often directly affect the therapeutic safety and efficacy of many drugs. However, there have been few studies on food-drug interactions involving transporters. Dietary polyphenols have been widely assumed to be beneficial to human health. Polyphenols are commercially prepared and used as functional foods. We report here for the first time that ferulic acid, which is widely used as a functional food, affects the transport of clinical agents. It is important to be aware of the potential of food-drug interactions and to act in order to prevent undesirable and harmful clinical consequences.     

Fractional extraction of compounds from grape seeds by supercritical fluid extraction and analysis for antimicrobial and agrochemical activities

White grape seeds were subjected to sequential supercritical fluid extraction. By increasing the polarity of the supercritical fluid using methanol as a modifier of CO(2), it was possible to fractionate the extracted compounds. Two fractions were obtained; the first, which was obtained with pure CO(2), contained mainly fatty acids, aliphatic aldehydes, and sterols. The second fraction, obtained with methanol-modified CO(2), had phenolic compounds, mainly catechin, epicatechin, and gallic acid. The fractions were bioassayed. Antimicrobial activities were checked on human pathogens, and a high degree of activity was obtained with the lipophilic fraction. Agrochemical activities on phytopathogenic fungi and activities on the etiolated wheat coleoptile bioassay were also checked. The more polar fraction was active in the latter bioassay.