赤眼蜂田间混合种群的分离、鉴定及其性比的确定

介绍了一种适用于赤眼蜂Trichogramma田间混合种群分离、鉴定及性比确定的简单而有效的方法,并据此方法成功地鉴定出4种自然寄生于小菜蛾卵的寄生蜂种类,拟澳洲赤眼蜂TrichogrammaconfusumViggiani、玉米螟赤眼蜂Trichogramma.ostriniaePangetChen、微突赤眼蜂TrichogrammaraioNagaraja及卷蛾分索赤眼蜂TrichogrammatoideabactraeNagaraja。     

A method for preparing freeze-clamped tissue samples for metabolite analyses

A rapid and simple method for preparing freeze-clamped tissue samples for metabolite determinations is described. Freeze-clamped rat heart tissue samples weighing from 0.8 to 1.0 g were homogenized directly in an Ultra-Turrax homogenizer for 60 s in 3.5 ml of ice-cold 0.6 M HClO4 without pulverizing them in liquid nitrogen. After centrifugation, the pellet was rehomogenized in the Ultra-Turrax homogenizer for 30 s in 1.5 ml of HClO4. Following a further centrifugation the extracts were combined and the pH was adjusted to 7.0 by adding 5 M K2CO3. The neutralized supernatant was used for the desired assays. The analyses of the tissue extracts obtained from isolated perfused rat hearts by the present method give similar results for different kinds of metabolites than those processed according to the previous classical method. Moreover, the values of the various parameters determined from the tissue extracts prepared according to the method described here are similar to the data reported in literature. The method can be readily applied to any other freeze-clamped tissue. The greatest improvement obtained is that the homogenization procedure can be accomplished easily and conveniently in about one-tenth of the time required for the earlier classical method without the time-consuming and unpleasant tissue grinding in liquid nitrogen.     

A new method for mite extraction from leaf samples

Debris often hampers the detection of mites in washed leaf samples. We describe in detail a method for the extraction of mites from leaf samples, based on the adherence of mite cuticles to liquid paraffin, at the interface of paraffin and ethanol in a so-called mite-counting channel. We demonstrate its efficacy by comparing the mite numbers in samples before and after extraction. We illustrate the method's reliability by extracting known numbers of a taxonomic variety of plant-inhabiting mites, manually added to mite-free debris: for 13 of the 15 taxa all mites were retrieved. This method can also be used to extract small non-mite arthropods such as scales, whiteflies, thrips, cicadellids, hymenopteran parasitoids and psyllids.     

A simplified paraffin embedding method for small botanical samples

Abstract

We present a simplified paraffin embedding method suitable for unsuberized or unlignified small botanical samples (diameter < 0.3 cm). Only 2 h are required to yield plant tissues embedded in paraffin for anatomical observation and molecular analysis. Our method achieved morphological preservation of cell structures and conservation of nucleic acids that were equivalent to the traditional protocol. Fourier transform infrared spectrometry showed that the degree of degradation of the cytoplasmic components (e.g., protein) resulting from our simplified protocol was similar to that of the traditional protocol. The DNA samples embedded using the simplified method was extractable and could be used for PCR analysis. The DNA quality was equivalent to that embedded using the traditional method.     

A method for preserving saliva samples at ambient temperatures

To facilitate biochemical and biopharmaceutical studies when cold storage is unavailable, we assessed the stability of saliva samples containing preservatives stored at room temperature over a 1-year period. Two preservative mixtures were evaluated: sodium benzoate and citric acid (P1), and ethyl and propyl paraben (P2). Saliva samples were spiked with acetaminophen (APAP) or antipyrine (AP) and stored in preservative-coated vials and examined for concentrations of APAP, AP, melatonin, and cortisol at regular intervals as a function of preservative type and storage duration. Samples were stored at room temperature or at -20 degrees C (positive control) and analyzed periodically for APAP and AP by high-performance liquid chromatography and for melatonin and cortisol by radioimmunoassay. The effectiveness of the preservatives was determined by calculating the value of samples stored at room temperature in terms of percent of control (-20 degrees C) values. P1 effectively maintained the stability of APAP (100%) and AP (100%) for 360 days at room temperature; concentrations in samples at room temperature on day 360 were comparable to those on day 01. P1 also effectively maintained melatonin (100%) and cortisol (95%) concentrations for 180 days at room temperature. P2 preserved AP and cortisol in saliva for 60 days, but APAP for only 14 days.     

A new method for clitoroplasty in male-to-female sex reassignment surgery.

Sex reassignment surgery has proved to be the best resolution for primary transsexuals. In 1980, Dr. Rubin stated that preservation of the glans to reconstruct the clitoris in male-to-female sex reassignment surgery gave good cosmetic and functional results. In his series, Rubin used the corpus spongiosum as the vascular pedicle of the neoclitoris, but urine leakage is a complication. From 1988 to the present, the dorsal portion of the glans penis with the dorsal neurovascular pedicle has been used here for clitoroplasty in nine male-to-female primary transsexuals. All neoclitorides survived well, with good preservation of light touch and sexual sensation. No urine leakage has occurred. Six patients who were followed reported sexual satisfaction.     

A rapid head-space method for ethyl alcohol determination in saliva samples

At neutral pH, tetraphenylboron dissolved in 3-heptanone extracts acetylcholine but not acetylcarnitine from aqucous media. At acid pH, both acetylcholine and acetylcarnitine are extracted. A pH curve of the efficiency of extraction of acetylcholine and acetylearnitine is given and the method of chemical synthesis of [1-acetyl-¹⁴C]carnitine is deseribed.     

A simplified method for determination of radioactive iron in whole-blood samples

For studies on iron absorption in man radioisotopes represent an easy and simple tool However, measurement of the orbital electron emitting radioiron, 55Fe, in blood is difficult and insufficiently described in the literature. The present study describes a relatively simple method for simultaneous determination of 55Fe and 59Fe in blood, using a dry-ashing procedure and recrystallization of the remaining iron. The detection limit of the method permits measurements of 0.1 Bq/ml blood thus allowing detection of less than 1% absorption from a 40 kBq dose, which is ethically acceptable in humans. The overall recovery of radioiron from blood is more than 90%, and the coefficient of variation, as judged by the variation in the ratio 55Fe/59Fe is in the order of 4%. Combined with whole-body counting of 59Fe and direct gamma-counting of 59Fe on blood samples, this method represents a sensitive method for studying the intestinal absorption of 55Fe and 59Fe in man and at the same time allows estimation of the amount of radioiron located in the vascular compartment.     

A new method for the detection of carotenoids in chlorophyll samples

Summary A new method is developed for the detection of carotenoids in chlorophyll samples.The typical colour curve of chlorophyll in 80 % methyl alcohol exhibits marked absorption in 690 to 610 and less absorption in 500 to 430. The colour curve of chlorophyll contaminated with carotenoids exhibits higher values in the region 530 to 430. If carotin is present, the band maximum in the region 530 to 430 is located at 500, and if xanthophyll is the impurity the band maximum is shifted to 430. On a comparison of the colour curve of the sample to be tested with that of the typical colour curve of chlorophyll the presence of carotenoids at once becomes evident, and carotin and xanthophyll are identified separately by the positions of the band maxima in the region 500 to 430.Carotenoids in as low a concentration as 0.05 % are detected by the new method described.     

A mixed two-stage method for detecting interactions in genomewide association studies

Genomewide association studies (GWAS) are being conducted to unravel the genetic etiology of complex diseases, in which complex epistasis may play an important role. One-stage method in which interactions are tested using all samples at one time may be computationally problematic, may have low power as the number of markers tested increases and may not be cost-efficient. A common two-stage method may be a reasonable and powerful approach for detecting interacting genes using all samples in both two stages. In this study, we introduce an alternative two-stage method, in which some promising markers are selected using a proportion of samples in the first stage and interactions are then tested using the remaining samples in the second stage. This two-stage method is called mixed two-stage method. We then investigate the power of both one-stage method and mixed two-stage method to detect interacting disease loci for a range of two-locus epistatic models in a case-control study design. Our results suggest that mixed two-stage method may be more powerful than one-stage method if we choose about 30% of samples for single-locus tests in the first stage, and identify less than and equal to 1% of markers for follow-up interaction tests. In addition, we compare both two-stage methods and find that our two-stage method will lose power because we only use part of samples in both two stages.     

A simple biosynthetic method for stereospecific resonance assignment of prochiral methyl groups in proteins

A new method for stereospecific assignment of prochiral methyl groups in proteins is presented in which protein samples are produced using U-[¹³C]glucose and subsaturating amounts of 2-[¹³C]methyl-acetolactate. The resulting non-uniform labeling pattern allows proR and proS methyl groups to be easily distinguished by their different phases in a constant-time two-dimensional ¹H-¹³C correlation spectra. Protein samples are conveniently prepared using the same media composition as the main uniformly-labeled sample and contain higher levels of isotope-enrichment than fractional labeling approaches. This new strategy thus represents an economically-attractive, robust alternative for obtaining isotopically-encoded stereospecific NMR assignments of prochiral methyl groups.     

A non-destructive method for identifying the sex of ant larvae

The differences between adult male and female ants are often striking and obvious, yet both sexes appear virtually identical at the larval stage. Current methods for determining larval sex rely on genetic analyses or histology, both of which require killing all larvae examined. Here, we describe a method for identifying larval sex in vivo based on visible differences in genital imaginal discs. Using a light microscope, clear differences in genital disc morphology were observed between male and female larvae of the ponerine ant Harpegnathos saltator. Next, we investigated whether this technique could be broadly applied within ants and found similar differences in genital discs between male and female larvae of Aphaenogaster cockerelli and Camponotus floridanus. Taken together, our results show that genital discs can be used as a reliable indicator of larval sex in species from at least three major ant subfamilies. This technique should facilitate research into topics where information about larval sex is required.