The isolation and characterization of 3-(2-carboxyethyl)cytosine following in vitro reaction of β-propiolactone with calf thymus DNA

The new adduct 3-(2-carboxyethyl)cytosine (3-CEC) was isolated following in vitro reaction of the carcinogen β-propiolactone (BPL) with calf thymus DNA. The structure of 3-CEC was confirmed by synthesis from BPL and dCyd. Reaction of BPL with cCyd (pH 7.0–7.5, 37°C) gave 3-(2-carboxyethyl)deoxycytidine (3-CEdCyd) (9% yield) and 3,N⁴-bis(2-carboxyethyl)deoxycytidine (3,N⁴-BCEdCyd) (0.6% yield). 3-CEdCyd and 3,N⁴-BCEdCyd were hydrolyzed (1.5 N HC1, 100°C, 2 h) to 3-CEC and 3,N⁴-bis(2-carboxyethyl)cytosine (3,N⁴-BCEC), respectively. The structure of 3-CEC was assigned on the basis of UV and NMR spectra and the electron impact (EI) mass spectra of 3-CEC and a tri-trimethylsilyl (TMS) derivative of 3 CEC as well as deuterated (d₂₇) tri-TMS derivative of 3-CEC. The structure of 3,N⁴-BCEC was assigned on the basis of UV spectra and the EI mass spectra of a tri-TMS derivative. EI and isobutane chemical ionization mass spectra of 3-methylcytosine (3-MeCyt) and a di-TMS derivative of 3-MeCyt were obtained and were helpful in deducing the structures of 3-CEC and 3,N⁴-BCEC. This is the first report of the alkylation by BPL of an exocyclic atom on a base in DNA. Compound 3,N⁴-BCEC was not detected in BPL-reacted calf thymus DNA. The relative amounts of 1-(2-carboxyethyl)adenine (1-CEA), 7-(2-carboxyethyl)guanine (7-CEG), 3-(2-carboxyethyl)thymine (3-CET) and 3-CEC isolated from BPL-reacted DNA following perchloric acid hydrolysis were 0.23, 1.00, 0.39 and 0.41 respectively, when the alkylation reaction was conducted in phosphate buffer at 0–5°C and pH 7.5 and 0.10, 1.00, 0.29 and 0.28 respectively when the reaction was conducted in H₂O at 37°C and pH 7.0–7.5.     

A comparison of the north-and southe european associations of Corallina officinalis L.

A comparison betweenCorallina officinalis associations from the northern Adriatic and the Icelandic coastal area is given regarding their floristic composition, zonal position and physionomy, as related to the environmental parameters and the different origins of the respective floras.     

Temperature-Sensitive Behavior of Paramecium caudatum

SYNOPSIS. The effect of temperature on the behavior of swimming cells of Paramecium caudatum has been investigated by photographic analyses of their tracks in uniform temperature, in temperature gradient, or in temperature changing with time. When the cells were placed in the temperature gradient, the frequency of discontinuous directional changes of cells swimming toward the optimal temperature, the temperature of the culture, was much lower than that of the cells swimming in the opposite direction. This difference in the frequency of directional changes explained the observed accumulation of the cells at - the optimal temperature. When the temperature was suddenly changed toward the optimum, a transient decrease of the frequency of directional changes was observed and when the temperature was changed in the reverse direction, a transient increase of the frequency was noted. This transient response to the temperature change was the origin of the dependence of the frequency of directional changes on the swimming direction in the temperature gradient. Finally, the relation between the magnitude of the transient response and the rate of the temperature change was derived.     

Adenosine cyclic monophosphate in potato tubers during storage at +10° and +2°

The levels of adenosine 3′: 5′ cyclic monophosphate in mature potato tubers (variety King Edward) stored at + 10°, for varying periods at + 2° and then after 28 days at + 2° transferred to + 10°, have been measured. The concentrations were very low (3–30 pmol/g fr. weight) and did not show marked changes with storage conditions.     

The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island wallaby (Macropus eugenii): development and hatching of the egg and the freeliving stages

Smales L. R. The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): development and hatching of the egg and the free living stages. International Journal for Parasitology7: 449–456. Labiostrongylus eugenii (Trichonematidae) occurs in the stomach of the Kangaroo Island Wallaby. Egg morphology is similar to that of other strongyloids. When incubated at 25°C embryogenesis is completed in about 30 h. An incomplete moult occurs within the egg, and larvae hatch at a sheathed second-stage $\text{43}\text{1}\text{2}\text{–83}\text{1}\text{2}$ h later. Development occurred at all temperatures between 2° and 37°C with an optimum about 25°C and an upper limit near 37°C. The hatching process is very rapid, taking about 2 min. It is signalled by increased larval activity followed by a change in shell permeability. The larva hatches at that pole of the shell which has become plastic.The sheathed second-stage larva measures 659.50 ± 22.54 μm by 27.98 ± 1.22 μm. Its internal structures are concealed by a mass of opaque granules which were demonstrated as neutral lipid by oil red O staining. A second incomplete moult at 3–4 days results in a doubly sheathed infective larva from which the lipid gradually disappears. The mouth never appears patent and the larvae neither feed nor grow but rather decrease in size with age. Optimal temperatures for larvae range between 15°–25°C with 37°C about the upper limit. The significance of this developmental pattern is discussed.     

Choline transport by synaptosomal membrane vesicles isolated from insect nervous tissue

Membrane vesicles derived from insect nervous tissue are capable of accumulating choline via a high affinity, carrier-mediated process with ion gradients as the sole driving force. The transport is strictly dependent on the presence of Na+ and Cl- in the medium and is stimulated by a membrane potential.     

Protein turnover in a psychrotrophic bacterium

Protein breakdown in pulse-labelled and longlabelled cells of Arthrobacter S ₁-55, a psychrotrophic bacterium, has been assessed at different temperatures. The temperature at which the cells were grown and labelled affected the breakdown of pulsed-labelled but not long-labelled proteins. Inhibitors of ATP synthesis inhibited proteolysis. Miscoding antibiotics stimulated the production of rapidly degradable proteins.     

Relations between elevated temperatures and fermentation behaviour of Kloeckera apiculata and Saccharomyces cerevisiae associated with winemaking in mixed cultures

One of the important factors affecting wine fermentation is temperature. The influence of elevated temperatures from 10 to 25 °C at 5 °C intervals on yeast growth and fermentation products were studied in mixed cultures of Kloeckera apiculata and Saccharomyces cerevisiae in grape juice. In the experiments carried out at 10 and 15 °C, K. apiculata grew and survived longer compared to trials conducted above 20 °C. In most cases, higher temperatures stimulated the production of higher alcohols but lowered the formation of esters and acetaldehyde. This revised version was published online in July 2006 with corrections to the Cover Date.     

An improved model for determining degree-day values from daily temperature data

Although using hourly weather data offers the greatest accuracy for estimating growing degree-day values, daily maximum and minimum temperature data are often used to estimate these values by approximating the diurnal temperature trends. This paper presents a new empirical model for estimating the hourly mean temperature. The model describes the diurnal variation using a sine function from the minimum temperature at sunrise until the maximum temperature is reached, another sine function from the maximum temperature until sunset, and a square-root function from then until sunrise the next morning. The model was developed and calibrated using several years of hourly data obtained from five automated weather stations located in California and representing a wide range of climate conditions. The model was tested against an additional data-set at each location. The temperature model gave good results, the root-mean-square error being less than 2.0 °C for most years and locations. The comparison with published models from the literature showed that the model was superior to the other methods. Hourly temperatures from the model were used to calculate degree-day values. A comparison between degree-day estimates determined from the model and those obtained other selected methods is presented. The results showed that the model had the best accuracy in general regardless of the season. Received: 25 October 2000 / Revised: 2 July 2001 / Accepted: 2 July 2001