Improvement of an enzyme-linked immunosorbent assay for equine herpesvirus type 4 by using a synthetic-peptide 24-mer repeat sequence of glycoprotein G as an antigen

To increase the sensitivity of an enzyme-linked immunosorbent assay (ELISA) for equine herpesvirus type 4 (EHV-4) that uses a 12-mer peptide of glycoprotein G (gG4-12-mer: MKNNPIYSEGSL) [4], we used a longer peptide consisting of a 24-mer repeat sequence (gG4-24-mer: MKNNPIYSEGSLMLNVQHDDSIHT) as an antigen. Sera of horses experimentally infected with EHV-4 reacted much more strongly to the gG4-24-mer peptide than to the gG4-12-mer peptide. We used peptide ELISAs to test paired sera from horses naturally infected with EHV-4 (n=40). gG4-24-mer ELISA detected 37 positive samples (92.5%), whereas gG4-12-mer ELISA detected only 28 (70.0%). gG4-24-mer ELISA was much more sensitive than gG4-12-mer ELISA.     

家蚕肾形卵的荧光差异显示分析

Liang等建立的mRNA的荧光差异显示法 (flourescentdifferentialdisplay,FDD)具有高效、安全、有用信息多等特点。利用这一方法在mRNA水平上探讨了肾形卵 (ki)与正常卵在初期胚胎阶段的基因表达 ,回收了差异片段 10条 ,并对其差异进行了验证。其中之一的片段序列分析表明为低分子量热激蛋白基因 (smallheatshockgene)。     

家蚕体形突变第2数珠蚕(mf-2)的遗传学研究

经X射线诱导发现的家蚕体形突变第2数珠蚕(mf-2)在形态上与数珠蚕(mf:12-39.8)相似。通过二者间的杂交实验表明,mf-2与mf为独立遗传;与各染色体的标记基因进行连锁分析,发现mf-2与第15染色体的眼纹全黑(bl:15-0.0)连锁,说明mf-2基因属于第15连锁群;初步测定mf-2与bl间的遗传距离为27.7 cM。     

Fetal heart rate monitoring from maternal body surface potentials using independent component analysis

The fetal heart rate is indispensable for monitoring the health of unborn cattle fetuses. To monitor the fetal heart rate, a method employing independent component analysis (ICA) to extract the fetal electrocardiogram (fECG) from potentials measured on the maternal body surface and composed of a mixture of the maternal ECG (mECG), fECG, baseline drift and noise is described. A mixing of the raw data was simplified using a linear time‐invariant model. To separate the fECG from the mECG, baseline drift, and noise, an ICA strategy was applied, using a hyperbolic tangent as the contrast function and treating mutual information with the minimization principle to find the optimum demixing matrix to derive the fECG from the measured signals. After the feasibility of this method was shown on simulated signals obtained by randomly mixing pure fECG, pure mECG, low frequency sinusoidal drift and noise, real signals from three cloned pregnant Holstein cows with 157, 177 and 224‐day gestation periods were used to verify the separation method. The results show that the fECG, mECG, low‐frequency sinusoidal drift and noise can be clearly segregated in simulations, and that the fECG, mECG, baseline drift and noise can be successfully derived from real signals. The ICA approach has great potential in effectively detecting the fECG from maternal body surface potentials.     

Initial design for a fish bioenergetics model of Pacific saury coupled to a lower trophic ecosystem model

A fish bioenergetics model coupled with an ecosystem model was developed to reproduce the growth of Pacific saury. The model spatially covers three different oceanographic spatial domains corresponding to the Kuroshio, Oyashio, and interfrontal (mixed water) regions. In this coupled model, three (small, large, and predatory) zooplankton densities which were derived from the lower trophic level ecosystem model were input to the bioenergetics model of saury as the prey densities. Although certain model parameters were imposed from other species’ bioenergetics, several model parameters were estimated from observational data specific to Pacific saury. The integrated model results reproduced appropriate growth rates of Pacific saury. Model sensitivities to water temperature and prey density are examined and observational methods to evaluate the model parameters are discussed.     

Mass spectrometric detection of proopiomelanocortin (POMC)-related peptides following molecular cloning of POMC cDNA in bigeye tuna Thunnus obesus

Several pituitary hormones, including adrenocorticotropin (ACTH), melanotropin (MSH) and β-endorphin, are generated from a common precursor protein, proopiomelanocortin (POMC). In fish, in addition to steroidogenesis of ACTH and melanogenesis of MSH, immunomodulating activity has been found in some POMC-related peptides. To investigate the functions of these peptides in the homologous system, it is necessary to establish a convenient detection method for the peptides. The present study aimed to establish a method for the detection of POMC-related peptides in bigeye tuna Thunnus obesus using a small amount of tissue sample, but not requiring peptide purification. We first determined the nucleotide sequence of tuna POMC cDNA. The cDNA was composed of 1084 base pairs (excluding the poly A tail) that encoded POMC consisting of 222 amino acids. We then fractionated an acid-acetone extract of one pituitary by reverse-phase high performance liquid chromatography (HPLC) and determined the molecular weight of each separated peptide by mass spectrometry. Consequently, we detected eight POMC-related peptides by comparing the values to the deduced amino acid sequence. Thus, the present study enabled the detection of POMC-related peptides from a small amount of tissue without the use of several purification steps.     

Efficacy of five commercial disinfectants and one anionic surfactant against equine herpesvirus type 1

We investigated the influences of various reaction conditions on equine herpesvirus type 1 (EHV-1) disinfection by 5 commercial disinfectants (3 quaternary ammonium compounds [QACs] and 2 chlorine-based disinfectants) and 1 anionic surfactant. QACs at their highest recommended concentrations had no virucidal effect on EHV-1 with a 10-min reaction time at 0°C or a 1-min reaction time at room temperature. Chlorine-based disinfectants achieved EHV-1 disinfection with a 10-min reaction time at −10°C or a 30-sec reaction time at room temperature. In the presence of 5% fetal bovine serum, QACs (except for benzalkonium chloride) showed more stable virucidal effects than did chlorine-based disinfectants. The virucidal effect of the anionic surfactant was almost equivalent to that of the QACs.