Chromatin structure. Further evidence against the existence of a beaded subunit for the 30-nm fiber

The size distribution of chromatin fragments released by micrococcal nuclease digestion of liver chromatin at various ionic strengths was examined. Below 20 mM ionic strength, gradient profiles with a peak centered at 6 nucleosomes are generated, whereas between 20 and 50 mM the peak is always centered on 12 nucleosomes, and above 50 mM ionic strength the 30-nm fiber becomes less accessible to the nuclease and there is a corresponding increase in the size distribution of fragments in the gradients. However, extensive digestions always give profiles with a peak of 12 nucleosomes as nuclease-resistant dodecamers accumulate. All of these observations are consistent with the winding of the 10-nm polynucleosome chain into a helical coil commencing at about 20 mM ionic strength. The helical turns are stabilized by histone H1 interactions between 20 and 50 mM ionic strength producing stable dodecamers. Above 50 mM ionic strength the coil condenses longitudinally and the profiles are consistent with a random attack of this fiber by the nuclease. Consequently it is not necessary to invoke the existence of a subunit bead to explain the profiles. We further define the conditions at which specific structural transitions take place and provide methodology for the preparation of chromatin at various levels of condensation.     

Chromatin structure. Evidence that the 30-nm fiber is a helical coil with 12 nucleosomes/turn

Sedimentation analysis has been used to compare the structure of 30-nm chromatin fibers, isolated and digested under conditions that maintain the native structure, with relaxed-refolded chromatin. The native chromatin fibers show sharp, ionic strength-dependent changes in sedimentation coefficient that are not apparent in relaxed-refolded fibers. The first transition at approximately 20 mM ionic strength reflects the organization of the 10-nm polynucleosome chain into a loose helically coiled 30-nm fiber. Between 20 and 60 mM ionic strength there is considerable interaction between nucleosomes within the coils to generate a stable helical array with 12 nucleosomes/turn. Above 60 mM ionic strength the helical coil continues to condense until it precipitates at ionic strengths slightly greater than those considered physiological, indicating that there is no end point in fiber formation. The data is incompatible with a solenoid model with 6 nucleosomes/turn and also rules out the existence of a beaded subunit structure.     

Some cytochemical studies on the cell surface ofAmphidinium carterae (Dinophyceae)

Summary The surface coat of the dinoflagellateAmphidinium carterae Hulburt was examined by fluorescence and transmission electron microscopy, using various fluorochromes and cationic dyes. The overall results showed cell-surface reactions typical of acid mucopolysaccharides. The cationic dye staining revealed an outer fine fibrillar layer (15–70 nm thick) overlying a dense anionic coat (40–60 nm thick) which appeared to thicken progressively with age. In general, the structure of the amphiesmal vesicles was similar to that previously described by other investigators. However, an acidic mucopolysaccharide layer was observed on the inner surface of these vesicles. Each of these structures is traversed by 1–3 pores and at least 2 types of extrusomes are formed, the spindle trichocysts and the mucocysts. Cell to cell adhesion through the surface coat was frequently observed. Evidence was also obtained for internalization of all the surface-coat markers used.This investigation forms part of a doctoral thesis submitted by the first author to the University of British Columbia, Vancouver, B.C.     

Mechanism of cadmium uptake by activated sludge

Summary The significance of metabolic activity in cadmium uptake by unacclimated activated sludge was studied. Below 30 mg/l cadmium in solution, biosorption was found to follow the Freundlich isotherm, which is the most common pattern for physico-chemical adsorption. More than 95% of total cadmium uptake was achieved within 5 min metal-sludge contact time. Biosorption increased strongly when the initial cadmium concentration in solution was raised from 10 to 100 mg/l, whereas in the same concentration range the metabolic activity of the sludge, as measured by respiratory activity and extracellular protein production, was very significantly inhibited. The addition of nutrients at low but significant levels failed to increase cadmium uptake in 2 h contact time, while in 24 h the addition of nutrients caused the biosorption to increase by only 5–10% without any significant growth of the biomass. Biosorption was found to increase with temperature between 5° C and 40° C, in correlation with an increase in the metabolic activity of the sludge. Pretreatment of the sludge with metabolic inhibitors (NaN₃ and UV rays) appeared to cause only a very slight decrease (5–10%) of biosorption. These results suggest that metabolic uptake of cadmium was low and that adsorption to the surface of the cells was the major mechanism of uptake.Offprint requests to: S. S. Sofer     

Tubificidae (oligochaeta) from the Ross Sea (Antarctica), with descriptions of one new genus and two new species

Three species,Torodrilus gelidus sp. nov. (subfamily Rhyacodrilinae),Rossidrilus terraenovae gen. et sp. nov. (Limnodriloidinae), and a second unnamed species of Limnodriloidinae, are reported from marine sediments in Terra Nova Bay, Ross Sea. Torodrilus gelidus is distinguished fromT. lowryi Cook, 1970 by its setal pattern (with few exceptions, both anterior and posterior setae are single-pointed in sexually mature specimens ofT. gelidus) and the morphology of its male protuberances (the latter folded over a midventral bursa in segment XI).Rossidrilus terraenovae is characterized by large diverticula attached to the oesophagus in the posterior part of segment IX, unpaired male and spermathecal pores, heavily muscular and entally ciliated atrial ampullae, elongate prostatic pads, and a deep, unpaired and muscular, copulatory sac. It is the first species of its subfamily to be described from Antarctic waters.     

Effect of osmotic stress on the growth of epicotyls of Cicer arietinum in relation to changes in the autolytic process and glycanhydrolytic cell wall enzymes

Simulation of drought by polyethylene glycol (PEG) inhibited elongation of epicotyls of Cicer arietinum L. cv. Castellana but had no effect on growth capacity since growth was restored once the inhibitory condition had been removed. The amount of proteins in the cell wall was correlated with the elongation of the epicotyls and decreased when elongation was inhibited. PEG-induced inhibition of elongation had different effects on the various glycanhydrolytic cell wall enzymes. Only α-galactosidase (EC 3. 2. 1. 22) seemed related to the lack of elongation, increasing its activity when elongation was inhibited. The β-galactosidase (EC 3. 2. 1. 23) and β-glucosidase (EC 3. 2. 1. 21) studied did not show changes in their specific activities during the inhibition of elongation. β-Galactosidase is responsible for the autolytic process in Cicer arietinum . This enzyme hydrolyzes specified linkages in the cell wall, releasing sugar constituents. Our present results show that β-galactosidase is not directly related with elongation because no changes could be observed during inhibition of elongation. The autolytic process is related with chemical processes taking place in the cell wall and preceding elongation of the epicotyls, i. e. the loosening process. Cell wall loosening is necessary for elongation to take place but elongation does not necessarily follow loosening if the osmotic conditions are unfavorable     

Isolation and partial characterization of a new mutant for sterol biotransformation in Mycobacterium sp.

Summary A new sterol biotransforming mutant was isolated from NRRL-B3683 Mycobacterium sp. after nitrosoguanidine mutagenesis. The mutant showed an enhanced ability to biotransform stigmasterol into 17-ketosteroids compared with the parental strain.     

Sterols and frost hardening of winter rape

Winter rape ( Brassica napus L., var. olifera cv. Górczanski) seedlings were exposed to hardening conditions and the content and composition of free sterols as well as the ratio of free sterols to total phospholipids were determined. There was a reduction in free sterol content in the leaves at the most advanced stage of hardening. The ratio of free sterol to total phospholipids was significantly reduced by hardening due to a decrease in the level of the former and an increase in that of the latter compounds. There was a negative correlation between this ratio and the temperature at which half of the seedlings died. Thus, adaptation of membranes to temperature takes place also at the level of sterol-phospholipid interactions. Exposing seedlings already hardened to freezing temperatures caused injury higher than 50%, and brought about a drastic increase in the level of free sterols and an elevation in the ratio of free sterols to phospholipids. The results are discussed in terms of a possible role of the molecular architecture of membranes in surviving at subzero temperatures.     

Growth and cell wall changes in rice coleoptiles

A fractionation of non-cellulosic sugars of Oryza sativa L. coleoptile cell walls was carried out and the composition of each fraction was studied during coleoptile growth.Percentages of fractions extracted with boiling water and with oxalate (pectic substances) were almost constant throughout development. An increase in the K II hemicellulosic fraction (extracted with 24% KOH) content, and a decrease in the K I hemicellulosic fraction (extracted with 10% KOH) were detected, when coleoptile growth finished.The percentage of glucose content in the K I hemicellulosic fraction was highest in young coleoptiles and lowest in old ones. Furthermore, a highly significant linear relationship between amounts of glucose and growth rate was obtained, while a inverse relationship between the amount of xylose and arabinose and growth rate was attained.Abreviations GLC gas liquid chromatography - IAA indole-3-acetic-acid - TFA trifluoroacetic acid - T_o minimum stress-relaxation time     

Strains of Colletotrichum coffeanum Noack Causing Coffee Berry Disease in Angola and Malawi with Characteristics Different to the Kenya Strain

Strains of Colletotrichum isolated from berries of Coffea arabica from Angola and Malawi were identified as C. coffeanum causing coffee berry disease (CBD). The isolates were highly aggressive to berries of the cultivar ‘Catimor’ and produced high numbers of acervuli and extremely large masses of pink conidia when cultivated in the dark. These strains are apparently different in aggressiveness and morphology to those described in the past from Kenya.