DNA loop domains in mammalian spermatozoa

The highly condensed and tightly packaged DNA of hamster spermatozoa was found to be organized into topologically constrained DNA loop domains attached at their bases to a nuclear matrix. The loop domains of the sperm nuclei differed from somatic cell loop domains from the same animal in two aspects. Sperm loop domains were 60% smaller than somatic cell loop domains, with an average DNA length of 46±7 kb in sperm as compared with 16±11 kb in brain. Secondly, unlike virtually all somatic cell DNA known which is negatively supercoiled, sperm DNA was devoid of detectable supercoiling. The presence of the loop domain structure in the highly condensed DNA of mammalian spermatozoa suggests that this motif is a fundamental aspect of eukaryotic DNA organization.     

Behavioral responses of Diabrotica adults to plant-derived semiochemicals encapsulated in a starch borate matrix

The concept of encapsulating semiochemicals into a starch matrix is being studied for potential use in corn rootworm (CRW) management programs. During 1987, experiments were conducted to determine: 1) If volatile plant-derived Diabrotica spp. attractants could be encapsulated in a starch borate matrix (SBM), and 2) If various SBM-semiochemical formulations would attract Diabrotica species over time in field corn. Chemical analyses of fresh SBM formulations indicated that indole, estragole, veratrole, phenylacetaldehyde, and trans-anethole were not retained during formulation but trans-cinnamaldehyde, Beta-ionone, 1,2,4,-trimethoxybenzene, eugenol and isugenol were successfully encapsulated. Encapsulated semiochemical formulations were made into 20 mesh granules, placed in Pherocon ® 1C traps that were tied to corn plants, and sampled for CRW adults every 4 days from 11 July to 8 September. Field data indicated that encapsulated semiochemicals were retained in the SBM for varying lengths of time and were released at rates attractive to CRW adults. A two-component mixture of trans-cinnamaldehyde and 1,2,4-trimethoxybenzene was the most effective formulation tested; however, no formulation was effective during corn silking and pollination. Although seasonal variation in CRW response could limit the usefulness of some plant-derived semiochemicals, the starch matrix concept may be useful as a delivery system for semiochemicals and may have potential as a tool that could be used in the development of new more biorational CRW management programs.

---

Résumé Le programme expérimental de 1987 était destiné à déterminer: 1) si les substances dérivées de végétaux et attractives pour Diabrotica pouvaient être encapsulées dans de l'amidon additionné d'acide borique; 2) si différentes formules attireraient les différentes espèces de Diabrotica dans un champ de maïs.L'indol, l'estragol, le vératrol, le phénylacétaldéhyde et le trans-anéthol n'ont pas été retenus, tandis que le trans-cinnamaldéhyde, la \-ionone, le 1,2,4-triméthobenzène, l'eugénol et l'isugénol ont été encapsulés avec succès dans des pièges attachés à des pieds de maïs (les détails techniques sont fournis). Les pièges ont été relevés tous les 4 jours du 11 juillet au 8 septembre. Les résultats montrent que les substances allélochimiques sont conservées dans la capsule pendant des durées variables et libérées à des concentrations attractives pour les Diabrotica adultes. Un mélange de trans-cinnamaldéhyde et de 1,2,4,-triméthoxybenzène a été la formule la plus efficace, à l'exception des périodes de formation des barbes et du pollen, où aucune formule n'a été attractive. Bien que la variation saisonnière des réactions de Diabrotica limite l'utilisation des substances allélochimiques d'origine végétale, la capsule d'amidon peut être employée pour libérer des substances allélochimiques et constitue un outil potentiel pour la mise au point d'une méthode plus rationnelle de lutte contre Diabrotica.

     

Glandular trichome exudate is the critical factor in geranium resistance to foxglove aphid

Resistance to the foxglove-aphid (Acyrthosiphon solani Kaltenbach) has been demonstrated in some inbred geranium lines (Pelargonium Xhortorum Bailey). To establish more definitively the cause/effect relationship between tall glandular trichome exudate and resistance, an intact plant bioassay was performed comparing a resistant plant line, a resistant plant line from which the tall glandular trichome exudate had been removed using a basic buffer solution, a susceptible line and a susceptible line treated with the buffer wash. After 5 days of isolation on the respective surfaces, the number of surviving adult aphids as well as the number of nymphs produced and remaining alive were determined. Aphids on the buffer washed, resistant line exhibited mortality and fecundity which was not significantly different from that produced by the susceptible line. In contrast, the untreated resistance line was clearly resistant with lower adult survival and fewer living nymphs. The tall glandular trichome exudate must therefore be a critical factor in geranium resistance to the foxglove aphid.

---

Zusammenfassung Widerstandsfähigkeit dem Fingerhut-Blattlaus (Acyrthosiphon solani Kaltenbach) gegenüber wurde in einigen durch Inzucht erzeugten Pelargonie-Linien (Pelargonium Xhortorum Bailey) gezeigt. Um das Verhältnis von Ursache und Wirkung zwischen dem hochgewachsenen glandulären Trichom-Exudat und Widerstandsfähigkeit genauer zu bestimmen, wurde eine Bio-Untersuchung an intakten Pflanzen unternommen. Dabei wurden eine widerstandsfähige Pflanzenlinie, eine widerstandsfähige Pflanzenlinie, von der das hochgewachsene glanduläre Trichom-Exudat durch eine basische Pufferlösung entfernt worden war, eine anfällige Linie und eine mit Pufferlösung behandelte Linie verglichen. Zwei erwachsene weibliche Blattläuse wurden fünf Tage durch ein engmaschiges Netz auf den zu untersuchenden dritten und vierten Knotenblättern eingesperrt. Bei jeder Pflanze wurde die Untersuchung an einem nichtbehandelten Blatt und an einem Blat, von dem das Exudat durch Waschen mit der Pufferlösung entfernt worden war, durchgeführt. Für jede Linie wurden fünf Pflanzen gebraucht, und der ganze Versuch wurde sechsmal wiederholt. Nach einer fünftägigen Isolierung auf den jeweiligen Oberflächen wurden die Blätter von der Pflanze entfernt, und sowohl die Zahl der überlebenden erwachsenen Blattläuse wie auch die der produzierten und noch am Leben gebliebenen Nymphen festgestellt. Mit einer niedrigeren Überlebensrate der Erwachsenen und weniger noch lebenden Nymphen war die nichtbehandelte widerstandsfähige Linie deutlich widerstandsfähig. Im Gegensatz dazu zeigten die Blattläuse auf der mit Puffer gewaschenen widerstandsfähigen Linie eine Sterblichkeit und Fruchtbarkeit, die nicht erheblich höher waren, als die auf der anfälligen Linie, was beweist, daß das Waschen mit der Pufferlösung den Widerstandsfaktor entfernt hatte. Das hochgewachsene glanduläre Trichom-Exudat muß deshalb ein kritischer Faktor in der Widerstandsfähigkeit gegen Pelargonie-Schädlinge sein.

     

Chromatin reorganization during emergence of malignant Friend tumors: early changes in H2A and H2B variants and nucleosome repeat length

Serial examination of five newly derived Friend murine tumors during early subcutaneous passages showed continuing changes in chromatin composition and structure over the first 10 to 20 passages followed by a period of stabilization over the subsequent 20 passages. These changes were reflected in a decrease in two major histone variants, H2A.1 and H2B.2, with a coordinate increase in histone variants, H2A.2 and H2B.1, and a changing nucleosome repeat length (NRL). The absolute values differed among the five tumors, but all five showed the same general direction of change. There was no obvious relationship among the NRL, H2A, and H2B histone variant values. A low H2A.1/H2A.2 ratio was found in Friend tumors of high malignant potential. Cell lines derived in vitro also showed directional changes in the H2A and H2B variants similar to those of their tumor cell parents, but with different kinetics. Our findings suggest that Friend tumor establishment is accompanied by an early period of chromatin reorganization marked by changes in several parameters of chromatin structure.     

Carbonyldiphosphonate, a selective inhibitor of mammalian DNA polymerase delta

Twenty-three pyrophosphate analogues were screened as inhibitors of proliferating cell nuclear antigen independent DNA polymerase delta (pol delta) derived from calf thymus. Carbonyldiphosphonate (COMDP), also known as alpha-oxomethylenediphosphonate, inhibited pol delta with a potency (Ki = 1.8 microM) 20 times greater than that displayed for DNA polymerase alpha (pol alpha) derived from the same tissue. Characterization of the mechanism of inhibition of pol delta indicated that COMDP competed with the dNTP specified by the template and was not competitive with the template-primer. In the case of pol alpha, COMDP did not compete with either the dNTP or the polynucleotide substrate. COMDP inhibited the 3'----5' exonuclease activity of pol delta weakly, displaying an IC50 greater than 1 mM.     

Solution structures of proteins from NMR data and modeling: alternative folds for neutrophil peptide 5

The structure of neutrophil peptide 5 in solution has recently been reported (Pardi et al., 1988). The structure determination was accomplished by using a distance geometry algorithm and 107 interproton distance constraints obtained from 2D NMR data. In each of the eight independent solutions to the distance geometry equations, the overall fold of the polypeptide backbone was identical and the root mean square (rms) deviation between backbone atoms of the superimposed structures was small (approximately 2.4 A). In this paper we report additional NP-5 structures obtained by using a new structure generation algorithm: a Monte Carlo search in torsion angle space. These structures have a large rms backbone deviation from the distance geometry structures (approximately 5.0 A). The backbone topologies differ in significant respects from the distance geometry structures and from each other. Structures are found that are pseudo mirror images of part or all of the fold corresponding to that first obtained with the distance geometry procedure. For small proteins, the problem of distinguishing the correct structure among pseudo mirror images is likely to be greater than previously recognized. When a set of test distance constraints constructed from a novel Monte Carlo structure is used as input in the distance geometry algorithm, the fold of the resulting structure does not correspond to that of the target. The results also demonstrate that the previously accepted criteria (the magnitude of the rms deviation between multiple solutions of the distance geometry equations) for defining the accuracy and precision of a peptide structure generated from NMR data are inadequate. An energetic analysis of structures corresponding to the different folding topologies has been carried out. The molecular mechanics energies obtained by minimization and molecular dynamics refinement provide sufficient information to eliminate certain alternative structures. On the basis of a careful comparison of the different trial structures with the experimental data, it is concluded that the NP-5 peptide fold which was originally reported is most consistent with the data. An alternative fold corresponding to structures with low energies and small total distance violations is ruled out because for this fold predicted NOEs are not observed experimentally.     

Effects of alpha-2 adrenoreceptor blockade by yohimbine on the hormonal response to hypoglycaemic stress in normal man

In order to evaluate the effect of alpha-2 adrenoreceptor blockade on the ACTH response to insulin-induced hypoglycaemia, six normal men were studied with and without yohimbine (30 mg p.o.) premedication. Despite a similar hypoglycaemic stimulus and significant suppression of the growth hormone response (P less than 0.05), no change was observed in basal or stimulated plasma ACTH, cortisol, arginine vasopressin (AVP) or prolactin responses following yohimbine. We conclude that alpha-2 adrenoceptor blockade with yohimbine does not significantly affect the ACTH response to hypoglycaemia in man.     

Functional bioassays utilizing zooplankton: a comparison

Functional zooplankton bioassays based on ingestion, reproduction and respiration are described, with methods for a new ingestion bioassay included. All bioassays are compared using three indices, including the variability of controls, the range of experimental responses, and a listing of contaminants causing inhibition/stimulation of response. The ingestion bioassay showed the greatest range of response, and was sensitive to pesticides, PCBs and heavy metals. It was also commonly characterized by a hormesis response. The reproduction bioassay showed the lowest variability, illustrated a reduced range of response, and was sensitive to nutrients and heavy metals. In one study, the respiration bioassay was sensitive only to PCBs.     

Quantitative genetics of seed size variation inPhlox

Summary Because seed size is often associated with survival and reproduction in plant populations, genetic variation for seed size may be reduced or eliminated by natural selection. To test this hypothesis we assessed genetic sources of variation in seed size in a population ofPhlox drummondii to determine whether genetic differences among seeds influence the size they attain. A diallel cross among 12 plants from a population at Bastrop, Texas, USA allowed us to partition variance in the mass of seeds among several genetic and parental effects. We found no evidence of additive genetic variance or dominance genetic variance for seed mass in the contribution of plants to their offspring. Extranuclear maternal effects accounted for 56% of the variance in seed mass. A small interaction was observed between seed genotype and maternal plant. Results of this study support theory that predicts little genetic variation for traits associated with fitness.     

The occurrence and distribution of certain polypeptides within the human carotid body

Summary Both carotid bodies from 26 patients coming to necropsy were fixed in 10% neutral buffered formalin and sections 4 m thick were stained for various peptides by use of the immunogold technique. The results show that the human carotid body contains met- and leu-enkephalin, substance P, vasoactive intestinal peptide (VIP), neurotensin and bombesin. The distribution of these six peptides within the carotid body differs. Thus met- and leu-enkephalin are both present predominantly within glomic chief cells but with a marked tendency to favour the dark variant of these cells. Substance P and VIP both show a weak immunoreactivity in comparison to the enkephalins and are present in all three variants of chief cell. Neurotensin shows the weakest immunoreactivity of all and is restricted to a few glomic chief cells in a minority of cases. Bombesin also shows a weak immunoreactivity in glomic chief cells but a strong reaction in glomic arteries and arterioles. In these vessels bombesin appears to be confined to smooth muscle cells in the media but we cannot say whether it is secreted by them or merely bound to receptor sites on their membranes. These findings are related to quantitative data on the concentration of peptides in the human carotid body from a previous paper with which we were associated.