Characterization of DNA methylation in the rat

In the rat, differentiation and cell proliferation both affect DNA methylation. We studied 5-methylcytosine at the inner cytosine of the sequence C-C-G-G, a common methylation site, using endonuclease MspI (which cleaves C-C-G-G- and C-^mC-G-G), and its isoschizomer HpaII (which cleaves only C-C-G-G). DNA from all tissues and cell lines studied was methylated at C-C-G-G, at levels ranging from 45 to 80%, but the methylation sites were not distributed uniformly. Our analysis suggests a model in which cells contain variable amounts of three DNA methylation states, averaging 30–40, 70–80 and 95–100% methylation, respectively. One biological parameter that alters methylation is the prolferative state of the cell. We observed that NRK, a non-transformed cell line, increased its DNA methylation from 45 to 67% when monolayer cultures became confluent and non-dividing. We also observed that a class of repetitive DNA was completely methylated in DNA from all sources except a transformed cell line.     

Entamoeba histolytica and Giardia lamblia: Growth responses to reducing agents

The growth responses of Entamoeba histolytica strains HM-1:IMSS and HK-9 to a variety of reducing agents were tested for one subculture in TYI-S-33 medium, prepared with no cysteine or ascorbic acid. Amoebae did not grow in this medium. Addition of l-ascorbic acid, d- or l-cysteine, or l-cystine each permitted the maximum growth observed. Dithiothreitol supported 68% maximum growth of HK-9 amoebae, but only 12% of HM-1. In contrast, growth of both strains was greatly diminished (0–13% growth) with 11 other compounds tested including glutathione, thiomalic acid, thioglycolic acid, and methionine. The growth responses of Giardia lamblia were similarly tested in TYI-S-33, as well as in TP-S-1 media. If l-cysteine was omitted from either medium, trophozoites did not grow, and eventually lysed. In TYI-S-33 medium, the requirement for l-cysteine was specific, whereas in TP-S-1 medium, other sulfhydryl compounds were partially effective and lower concentrations of l-cysteine satisfied the requirement. Ascorbic acid or l-cystine alone was totally ineffective; however, in combination, 30 to 60% of maximum growth was achieved. Once added to either medium, cysteine was rapidly oxidized. Amino acid analysis of the growth media revealed that the broth components of TP-S-1 medium contained 2.8 mM and TYI-S-33 broth 2.1 mM endogenous levels of cysteine (or half-cystine), with an additional 3 mM contributed by 10% serum.     

Contribution a l'etude limnologique du lac Pavin (France). II: Relations entre parametres abiotiques-phyto et zooplancton dans la zone 0–20 metres

At 1197 m in the French Massif Central, lake Pavin has a volcanic origin. It is a meromictic lake of 44 ha area and 98 meters depth.Zooplancton samples (250 liters) were collected bimonthly with a water pump from May 1973 to April 1974 at six depths from surface to 20 meters.Countings were made to study the dynamics of two Cladocera: (Daphnia longispina and Ceriodaphnia quadrangula), two copepode populations, the calanoide Acanthodiaptomus denticornis and the cyclopoide Cyclops abyssorum. For the last two species, males, females, ovigerous females, Nauplius larvae and each of the five copepodite stages were differentiated.If Cladocera have a period of development limited from June to September for Ceriodaphnia and from June to December for Daphnia, copepods are presents all around the year with two principals periods of reproduction. The first during spring (March–April), the second during fall (October–November). If this second generation presents an important development for Cyclops abyssorum, that is not the same for Acanthodiaptomus denticornis, in which species nauplius larvae died in winter. The synthetical mathematical model realized by Principal Components Analysis illustrates the relationships between phyto-and zooplancton and bio-physico-chemicals parameters and specially the fact that, in this lake, the highest values of phytoplanctonic primary production and activity coefficient (Production/Standing crop) are found when herbivorous zooplancton presents the more important development and nutrients their lowest concentrations.

     

Identification of two nuclear polyhedrosis viruses from the cabbage moth, Mamestra brassicae (Lepidoptera: Noctuidae)

Two nuclear polyhedrosis viruses from the cabbage moth Mamestra brassicae found in two geographical areas in Europe have been characterized and compared. These two virus isolates have similar biological activities and have the same host range. The two M. brassicae nuclear polyhedrosis viruses can be distinguished by restriction endonuclease analysis of their DNA. They appear to be distinct but related virus strains.     

A new variant of Autographa californica nuclear polyhedrosis virus

A variant of the baculovirus, Autographa californica nuclear polyhedrosis virus, has been isolated in Idaho during an epizootic disease in a field population of A. californica. Genotypic characterization indicates that the virus is distinct from variants previously characterized. Analysis of five clones, derived by plaque purification in cell culture, indicates relative homogeneity of the original virus isolate. Further exploration of the factors involved in natural genetic variability of baculoviruses is appropriate.     

The hypothalamo-hypophysial system in the ground squirrel,Citellus erythrogenys Brandt

Summary Monthly observations of the Gomori-positive hypothalamo-hypophysial neurosecretory system (HHNS) of the ground squirrel, Citellus erythrogenys Brandt, were carried out light microscopically using several quantitative methods. From the beginning of hibernation, formation of neurosecretory material (NSM) in the neurosecretory cells (NSC) progressively decreases and release of neurohormones from the HHNS is almost fully inhibited. A maximal accumulation of NSM in the perikarya of the NSC and in the posterior pituitary (PP) is found in December. By this time the volume of the cell nuclei and nucleoli is at a minimum. Signs of activation of the HHNS appear and become more conspicuous as the time of arousal from torpor approaches. The amount of NSM in the NSC and the PP decreases simultaneously with the increase in volume of the NSC. Hyperemia and activation of glial elements is visible throughout the HHNS. The morphological signs of activation reach their peak in March. After reproduction is completed (April to beginning of May), the NSC and the PP are almost devoid of NSM. Beginning with June and during the summer and autumn months a progressive accumulation of NSM in the NSC and the PP parallels gradual diminution in the volume of the NSC structures and the glial cells. Mechanisms and effector pathways by which the HHNS influences seasonal adaptation of the organism and reproduction are discussed.     

Parameter identification of the calvin photosynthesis cycle

Summary This article is concerned with the determination of kinetic parameters of the Calvin photosynthesis cycle which is described by seventeen nonlinear ordinary differential equations. It is shown that the task requires dynamic data for several sets of initial conditions. The numerical technique is based upon an algorithm for non-linear optimization and Gear's numerical integration scheme for stiff systems of differential equations. The sensitivity of the parameters to noise in the data is tested with a method adapted from Rosenbrook and Storey. A preliminary set of parameters has been obtained from a preliminary set of experimental data. The numerical methods are then tested with synthetic data derived from these parameters. The mathematical model and the results obtained in the simulation are used as an aid in designing new experiments.