Clustering of null mutations in the EcoRI endonuclease

EcoRI endonuclease mutants were isolated in a methylase-deficient background following in vitro hydroxylamine mutagenesis of plasmid pKG2 (Kuhn et al.: Gene 44:253-263, 1986). Mutants which survived high-level endonuclease expression (IPTG induction) were termed null mutants. Sixty-two of 121 null mutants tested by Western blot contained normal levels of endonuclease cross-reacting protein. The complete endonuclease gene was sequenced for 27 null mutants. This group was found to consist of 20 single base-change missense mutations, 6 double mutations, and 1 triple mutation. Ten of the 20 single mutations were clustered between residues 139 and 144. When examined with respect to the structure of the EcoRI-DNA complex (McClarin et al.: Science 234:1526-1541, 1986), these alterations were found to fall predominantly into two classes: substitutions at the protein-DNA interface or substitutions at the protein-protein (dimer) interface. Protein from several of the mutants was purified and sized by using HPLC. Wild-type EcoRI endonuclease and protein from three of the DNA interface mutations (Ala139----Thr, Gly140----Ser, Arg203----Gln) appeared to be dimeric, while protein from subunit interface mutations (Glu144----Lys, Glu152----Lys, Gly210----Arg) migrated as monomers.     

The effect of a chemical deterrent, released from the frass of caterpillars of the garden pebble moth, on cabbage root fly oviposition

The cabbage root fly, Delia radicum (L.), was deterred from laying eggs on cauliflower plants that had been sprayed with a suspension of the frass of caterpillars of the garden pebble moth, Evergestis forficalis (L.). Polar extracts of the frass deterred oviposition irrespective of the cruciferous plant species on which the caterpillars had been feeding. Non-polar extracts of the frass had no effect. Spraying plants with macerates from Brassica leaves stimulated fly oviposition whereas spraying plants with macerates from garlic mustard leaves deterred fly oviposition. Macerates from the leaves of all other plants tested had no effect. In field experiments the deterrent effect persisted 2–3 days after leaves were sprayed with frass extracts. Plants infested with feeding caterpillars and contaminated with only a few discrete frass pellets were as deterrent to the fly as those sprayed with frass suspensions.

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Résumé D. radicum a évité de pondre sur des pieds de chou-fleur, qui avaient été pulvérisés avec une suspension d'excréments d'E. forficalis. Les extraits polaires de ces excréments empêchent la ponte quelle que soit l'espère de crucifère sur laquelle les chenilles se sont alimentées. Les extraits non polares sont sans effet. Dans la nature, l'action dissuadante persiste 2 à 3 jours après la pulvérisation des feuilles avec les extraits d'excréments. La pulvérisation tous les deux jours a maintenu l'action dissuadante. Des plantes contaminées par des chenilles en train de s'alimenter et souillées par quelques crottes sont aussi dissuadantes pour la mouche que celles pulvérisées avec une suspension d'excréments.

     

Connective tissue influences on the expression of epithelial cell-surface antigens

Summary Adult mice were found to show regional variation in the epithelial expression of some molecules of the blood-group antigen series. To investigate connective tissue influences on such differences, heterotypic recombinants of epithelia and connective tissues from various regions were prepared and examined using monoclonal antibodies directed against bloodgroup antigens H and Le^y. The results indicate that epithelia may maintain a preexisting regionally specific pattern following recombination but that, in some recombinant matches, the connective tissue is capable of signalling redirection of the pattern of expression towards that typical of the epithelium with which it is normally associated.This work was supported by NIH-NIDR RO1-DEO-5190     

The effects of gregarine parasites on longevity, weight loss, fecundity and developmental time in the field crickets Gryllus veletis and G.pennsylvanicus

ABSTRACT. 1. Gregarines, debilitating gut parasites, significantly reduced longevity and increased weight loss in the field cricket Gryllus pennsylvanicus Burmeiser when adults were kept on suboptimal diets, but in G.pennsylvanicus and G.veletis (Alexander and Bigelow) fed ad libitum , weight change, longevity, and fecundity were not significantly different in infected and control individuals.
2. Adults harbouring gregarines at eclosion took significantly longer to develop from the second and third instars than did uninfected adults.
3. Longevity of nymphs kept in the laboratory without food was decreased in infected individuals compared with uninfected ones, but not significantly so.
4. The evolutionary and ecological implications of nonlethal parasites are discussed.     

Immunofluorescent localization of collagen types I,III and IV,fibronectin, laminin,and basement membrane proteoglycan in developing mouse skin

Summary The distribution of various extracellular matrix components was studied in frozen sections of embryonic (14–18 days) and early postnatal (birth and 4 days post parturn) dorsal mouse skin using monospecific antibodies and indirect immunofluorescence. Basement membrane zone components — type IV collagen, laminin and heparan sulphate proteoglycan — were found to be uniformly and unchangingly distributed along the dermal-epidermal junction. In contrast, the distribution of interstitial matrix components — types I and III collagen, and fibronectin — was heterogeneous and varied with the stages of hair development. Collagens became sparse and were eventually completely removed from the prospective dermal papilla and from a one-cell-thick sheath of dermal cells around hair buds. They remained absent from the dermal papilla throughout hair organogenesis. Fibronectin was always present around dermal papilla cells and was particularly abundant along the dermal-epidermal junction of hair rudiments, as well as underneath hair buds. In contrast, in interfollicular skin, collagens accumulated in increasing density, while fibronectin became progressively sparser. It thus appears that interstitial collagens and fibronectin are distributed in a manner which is related to hair morphogenesis. In morphogenetically active regions, collagen density is low, while that of fibronectin is high. Conversely, in histologically stabilized zones, collagen is abundant and fibronectin is sparse. This microheterogeneous distribution of interstitial collagens and of fibronectin might thus constitute part of the morphogenetic message that the dermis is known to transmit to the epidermis during the development of skin and of cutaneous appendages.     

Evolutionary steps of ecophysiological adaptation and diversification of ruminants: a comparative view of their digestive system

Summary A review is made of the ruminant digestive system in its morphophysiological variations and adaptations relating to foraging behaviour, digestive physiology, to interactions between plants and ruminants and to geographic and climatic diversity of ruminants' ecological niches. Evidence is provided for evolutionary trends from an extreme selectivity mainly for plant cell contents and dependence upon a fractionated fore- and hindgut fermentation, to an unselective intake of bulk roughage subjected to an efficient plant cell wall fermentation, mainly in the forestomachs. The review is based on detailed comparative morphological studies of all portions of the digestive system of 65 ruminant species from four continents. Their results are related to physiological evidence and to the classification of all extant ruminants into a flexible system of three overlapping morphophysiological feeding types: concentrate selectors (40%), grass and roughage eaters (25%) and intermediate, opportunistic, mixed feeders (35%). Several examples are discussed how ruminants of different feeding types are gaining ecological advantage and it is concluded that ruminants have achieved high levels of digestive efficiency at each evolutionary stage, (including well-documented seasonal adaptations of the digestive system) and that ruminant evolution is still going on. Deductions made from the few domesticated ruminant species may have, in the past, biased scientific evaluation of the free-ranging species' ecology. The main threat to a continuous ruminant evolution and diversity appears to be man's neglect for essential ecological interactions between wild ruminants and their specific habitats, which he alters or destroys.Abbreviations bw body weight - CS concentrate selector - DFC distal fermentation chamber (distended caecocolon) - GR grass and roughage eater - IM intermediate (mixed) feeder - PFC proximal fermentation chamber (ruminoreticulum/forestomachs) - RR Ruminoreticulum - SCFA Short-chain fatty acis (acetic, butyric, propionic acid set free by rumen bacteriae) - SE Surface enlargement (of absorptive mucosa) Supported by German Research Community grant DFG Ho 273/6Dedicated to Professor Dr. Dr. h.c. Dietrich Starck on the occasion of his 80th birthday     

Relationships between some genetic parameters and test environments in open-pollinated families of Pinus elliottii in South Africa

Summary Growth traits, stem form and branch characteristics in three series of well-replicated trials, comprising open-pollinated families of Pinus elliottii Engelmann, were analysed to investigate whether the absolute value of the component of variance for family effect and its relative value (which is equivalent to one-quarter of heritability on individual values) were related, with trial averages of the traits studied taken as productivity indices of the sites tested. The first series, called the foreign selection series, consisted of four trials comprising open-pollinated families from single clones from a Florida seed orchard and from selected trees in Queensland, Australia. Two trials were established in the eastern Transvaal, at Tweefontein and Frankfort State Forests (SF), and two in Zululand, at Dukuduku and KwaMbonambi SF. The second series, called the Zululand selection series, consisted of open-pollinated families from single trees selected in Zululand that were compared in three trials: in the eastern Transvaal at Wilgeboom SF, in Zululand at Dukuduku SF and in the southern Cape at Lottering SF. The third series, called the South African selection series, consisted of open-pollinated families from single trees selected in the different forest areas of South Africa and compared in four trials: two trials in the eastern Transvaal at Tweefontein and Mariti SF and two in the Natal province at Dukuduku and Weza SF. Some positive and linear relationships between absolute sizes of the variance component for family and trial average for all traits studied, except stem form, were discernible for the foreign selections and for the Zululand selections. The South African selections presented a more erratic pattern of variation. However, when the origin of the genetic material was disregarded, the positive trends were undeniable. Curvilinear relationships between relative values and site averages were discernible, when the origin of the genetic material was disregarded. Further research is needed in order to confirm the suggested effect of site on genetic expressivity of open-pollinated families of slash pine grown in South Africa.     

Application of the Fröhlich theory to the modelling of rouleau formation in human erythrocytes

The details of Fröhlich's theory and some recent experiments on the rouleau formation of human erythrocytes which exhibit a strong interaction that appears to satisfy the prerequisites of the Fröhlich theory, are summarized. To verify whether the Fröhlich theory of long-range coherence in biological systems is applicable to the phenomenon of rouleau formation in human erythrocytes, the interactions between erythrocytes are modelled as those between two large, coupled oscillating dipoles. Relevant expressions for the resonant long-range and the van der Waals interaction are then derived. Using the available numerical data, the eigenfrequencies and the interaction energies corresponding to the experimental conditions are then derived. In the range of postulated frequencies (10¹¹–10¹² Hz) the effective interaction coefficient due to the resonant long-range forces is, indeed, found to agree with its experimental value of 3.0. However, the same value of can also be achieved through the ordinary van der Waals interactions between dipoles oscillating at lower frequencies. It is concluded that the resonant long-range interaction between erythrocytes may be responsible for the onset of rouleau formation. However, other mechanisms cannot be ruled out at this stage, especially since the Fröhlich mechanism requires a number of unconfirmed preconditions.     

The effect of low- and high-density lipoprotein cholesterol on steroid hormone production and ACTH-induced differentiation of rat adrenocortical cells in primary culture

Summary The choroid plexus consists of the choroidal epithelium, a derivative of the neural tube, and the choroidal stroma, which originates from the embryonic head mesenchyme. This study deals with epithelio-mesenchymal interactions of these two components leading to the formation of the organ. Grafting experiments of the prospective components have been performed using the quail-chicken marker technique. Prospective choroidal epithelium of quail embryos, forced to interact with mesenchyme of the body wall of chicken embryos, gives rise to a choroid plexus showing normal morphogenesis and differentiation. The choroidal epithelium induces the differentiation of organtypical fenestrated capillaries, which are highly permeable to intravenously injected horseradish peroxidase. The choroidal epithelium of the grafts constitutes a blood-cerebrospinal fluid barrier. On top of the choroidal epithelium, there are epiplexus cells displaying a typical ultrastructure. The experimental results show that these cells do not originate from the transplanted neural epithelium. Prospective choroidal stroma of chicken embryos does not exert a choroid plexus-inducing influence upon a quail embryo's neural epithelium isolated from parts of the brain that normally do not develop a choroid plexus. The experiments show that the choroidal epithelial cells are determined at least three days before the first organ anlage is detectable.This work was supported by the Deutsche Forschungsgemeinschaft (grant Ch 44/7-1)     

Membrane-damaging action of ricin on DPPC and DPPC-cerebrosides assemblies

Perturbations induced by a toxic lectin (ricin) on lipid organisation of model membranes prepared with DPPC and DPPC-cerebrosides mixtures have been analysed by Raman and infrared spectroscopy, two powerful and non-invasive methods. Our approach involves the observation of changes in the vibrational spectra of liquid multilayers in the PO ₂ ^- , C=0 and CH₂ spectral regions for two lipid: ricin molar ratios (225:1, 75:1).The interfacial and polar regions of the multilayers, analysed by FTIR, appear to be perturbed by the protein. With both kinds of membranes, ricin mainly perturbs the C=0 ester groups of the sn-2 acylchain of DPPC. In the PO ₂ ^- stretching region, the frequency shifts are correlated with changes in polar group hydration.In the hydrophobic core of the multilayer membrane studied by Raman spectroscopy, the interaction of ricin is associated with changes in lipid packing. These perturbations depend upon the lipid composition of the membrane. With DPPC membranes, an affect is detected at temperatures lower than T _m .It corresponds to a decrease of the lipid ordering. With DPPC-cer membranes, the protein increases the acylchain packing order regardless of the temperature of the experiments (10°C<T<75°C). No perturbation of T _m is observed after addition of ricin to either DPPC or DPPC-cer membranes.The different perturbations detected by Raman and FTIR suggest that ricin mainly interacts with the interfacial domains of the membranes.