Allium cepa L. Cultivars from Four Continents Compared by Flow Cytometry show Nuclear DNA Constancy

In 1965 Van't Hof estimated the nuclear DNA amount of an unidentifiedAllium cepa L. cultivar as 2C = 33.55 pg (Experimental CellResearch39: 8–58). This value has been adopted by commonusage as the main calibration standard for angiosperm DNA C-valueestimations. However, different cultivars have been used whileassuming species DNA C-value constancy. Surprisingly this assumptionhas never been tested. A. cepa is an outbreeder with telomericheterochromatic segments, so intraspecific variation in C-value,possibly correlated with environmental factors as seen in Zeamays L., might be expected. We used laser flow cytometry tocompare nuclear DNA amounts in roots of six A. cepa cultivarsused as calibration standards or from different environments.Tissues from one cultivar, or similar volumes of tissue fromtwo cultivars, were run and the variance between nuclei in 2Cpeaks compared. Only one shoulderless 2C peak was seen for allpairs of co-chopped cultivars. Thus, no large differences inC-value between cultivars from different environments were found.Moreover, comparing cultivars run singly or as pairs showedno evidence for increased variation in 2C peaks in the latter,and hence of critical differences in DNA amounts between ‘AilsaCraig’ and another cultivar. Such variation was insufficientto make their use as alternative calibration standards, or thepractice of imputing Van't Hof's original C-value estimate tothem, unacceptable for most practical purposes. Given the mechanismsknown which can generate genome size variation, the degree ofconstancy in DNA C-value found seems remarkable. Copyright 2000Annals of Botany Company Allium cepa, onion cultivars, calibration standards, DNA C-value constancy, flow cytometry     

1-Alkyl-3-amino-5-aryl-1H-[1,2,4]triazoles: novel synthesis via cyclization of N-acyl-S-methylisothioureas with alkylhydrazines and their potent corticotropin-releasing factor-1 (CRF(1)) receptor antagonist activities.

Cyclizations of alkylhydrazines with N-acyl-S-methylisothioureas, readily synthesized from acyl chlorides, sodium thioisocyanate, dialkylamines then methyl iodide in a one-pot reaction, gave 1-alkyl-3-dialkylamino-5-phenyltriazoles 7 as major products. The regioisomers were assigned through the use of NOE NMR experiments. While bearing a N-bis(cyclopropyl)methyl-N-propylamino group, this series of compounds shows very good binding affinity on the human CRF₁ receptor. Among them, 1-methyl-3-[N-bis(cyclopropyl)methyl-N-propylamino]-5-(2,4-dichlorophenyl)-1H-[1,2,4]triazole 7a had the best binding affinity for the CRF₁ receptor (K_i=9 nM).     

Defoliation-induced enhancement of total aboveground nitrogen yield of grasses

We conducted an experiment in a northern mixed-grass prairie at Wind Cave National Park, South Dakota, USA to evaluate the effect of defoliation frequency on aboveground net primary production (ANPP), shoot nitrogen concentration, and aboveground N yield of graminoids. ANPP was significantly reduced at weekly and biweekly defoliation frequencies, but unaffected relative to unclipped controls at monthly and bimonthly frequencies. By contrast, clipping at all frequencies increased shoot N concentration above that of controls, and this increase was greatest at monthly or more frequent defoliations. Total aboveground N yield and potential N yield to grazers were greatest at intermediate (bimonthly to biweekly) frequencies. We suggest that grazers may maximize their nutritional status in this system by periodically regrazing areas at frequencies near the approximately monthly optimum that we observed.     

Use of Radiation Hybrid panels to map genetic loci

The mapping of genetic loci within organisms has been accelerated by the advent of Radiation Hybrid (RH) panels. These panels are available for humans and non-humans including mice, baboon, rat, and canine. This article contains a general protocol for the use of the Genebridge 4 whole genome RH panel to map a human locus. This protocol may also be adjusted to suit the other RH panels currently available.     

HYLAS: program for generating H curves (abstract three-dimensional representations of long DNA sequences)

The computer program HYLAS generates from a standard DNA lettersequence a three-dimensional space curve (H curve) which embodiesthe entire information content of the original nucleotide sequence.The program can display H curves either as two-dimensional (frontand side view) projections or as stereo-pair images. The curvescan be marked at specific nucleotide locations, annotated, rotatedfor observation from any viewing angle, and manipulated forconvenient side-by-side comparisons. Unlike the cumbersome lettersequences, H curves can be drastically condensed in size withoutlosing their ability to reflect the global nucleotide-distributionpattern of the entire DNA sequence. Often, biologically importantloci can be visually identified on the H curves. HYLAS is writtenin FORTRAN with separate mainframe (IBM- VM/CMS) and microcomputer(MS-DOS) versions. It uses the Tektronix-TCS library of graphicsubroutines. Received on October 24, 1988; accepted on July 15, 1989     

Simple reversed-phase high-performance liquid chromatography quantitation of ganciclovir in human serum and urine

A fast, simple, and cost-effective HPLC method for the quantitation of the antiviral drug ganciclovir is described. The serum samples are extracted with perchloric acid and neutralized with potassium phosphate buffer, and urine samples are diluted with distilled water. A reversed-phase column with isocratic elution by 15 mM potassium phosphate buffer (pH 2.5) containing 0.25% acetonitrile is used to separate ganciclovir; quantitation is by UV absorbance at 254 nm. Total turnaround time is 22 min; more than 3000 samples can be run on a single column without loss of peak quality. The limit of quantitation is 0.05 μg/ml. Recoveries varied from 91 to 10% with coefficients of variation ranging from 0.387 to 7.95%.     

Estimating recent growth in the cuttlefish Sepia officinalis: are nucleic acid-based indicators for growth and condition the method of choice?

A laboratory calibration study was undertaken with juvenile Sepia officinalis (80-85 g initial wet weight) to investigate the effects of different food rations and different starving intervals on RNA/dry weight (DW) ratios and RNA/DNA ratios in cephalopod mantle muscle at two different temperatures. The digestive gland index was also used as an additional indicator of recent growth. High food rations and low temperature went along with high RNA/DW ratios and high RNA/DNA ratios. Starving resulted in a linear decline in growth performance and a concomitant decrease in RNA/DW and RNA/DNA ratio, with RNA/DNA ratios representing the growth data better. RNA/DNA ratios decreased faster at higher temperatures. A fluorimetric assay for nucleic acid analysis was optimized for cephalopod mantle tissues and yielded reproducible RNA/DNA ratios with a relative variance below 10%. Thus, it may be possible to use this estimator of recently encountered feeding regime for the evaluation of mortality rates of early teuthid paralarvae to eventually support stock management. Also, log relative digestive gland weight showed a strong relationship with starving time, but, surprisingly, not with temperature. Data from the two temperatures analyzed could be combined to form a common regression line of relative digestive gland index with starving time. This indicator for recent growth might be especially suitable for large specimens with a well-developed digestive gland.     

In vitro metabolism or testosterone by breast microcysts

Breast microcysts are considered to be a normal findings in the adult female breast without any increased risk of developing carcinomatous change. Breast cysts fluid contains steroid but not studies have been reported on the ability of breast microcysts to metabolise steroid hormones. It was, therefore, the aim of this study to identify the metabolites formed on incubation of radiolabelled testosterone with microcysts. In all instances dihydrotestosterone and androstenedione were formed. Oestrogens were not identified. Tis study, therefore, provides evidence for th presence of 5-alpha-reductase and 17-oxidoreductase enzyme systems in breast microcysts.