Quantification of the calcium-induced secondary structural changes in the regulatory domain of troponin-C.

The backbone resonance assignments have been completed for the apo (1H and 15N) and calcium-loaded (1H, 15N, and 13C) regulatory N-domain of chicken skeletal troponin-C (1-90), using multidimensional homonuclear and heteronuclear NMR spectroscopy. The chemical-shift information, along with detailed NOE analysis and 3JHNH alpha coupling constants, permitted the determination and quantification of the Ca(2+)-induced secondary structural change in the N-domain of TnC. For both structures, 5 helices and 2 short beta-strands were found, as was observed in the apo N-domain of the crystal structure of whole TnC (Herzberg O, James MNG, 1988, J Mol Biol 203:761-779). The NMR solution structure of the apo form is indistinguishable from the crystal structure, whereas some structural differences are evident when comparing the 2Ca2+ state solution structure with the apo one. The major conformational change observed is the straightening of helix-B upon Ca2+ binding. The possible importance and role of this conformational change is explored. Previous CD studies on the regulatory domain of TnC showed a significant Ca(2+)-induced increase in negative ellipticity, suggesting a significant increase in helical content upon Ca2+ binding. The present study shows that there is virtually no change in alpha-helical content associated with the transition from apo to the 2Ca2+ state of the N-domain of TnC. Therefore, the Ca(2+)-induced increase in ellipticity observed by CD does not relate to a change in helical content, but more likely to changes in spatial orientation of helices.     

Brinckochrysa scelestes [Neur.: Chrysopidae] as predator ofMyzus persicae [Hom.: Aphididae] on tobacco

Laboratory and field studies were conducted to study the suitability and efficacy ofBrinckochrysa scelestes Banks as a predator onMyzus persicae Sulzer in tobacco fields. Laboratory bred 2nd instar larvae ofB. scelestes at 6 per tobacco plant reduced the aphid population to the extent of 78% in 2 weeks. The movement of the predatory larvae on the tobacco plant and leaf was not at all hampered by the glandular trichomes and their secretions. The larvae moved freely and fed well and their pupae were recovered from all over the plant.

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Résumé On étudie en laboratoire et en champ de tabac la valeur et l'efficacité deBrinckchrysa scelestes Banks comme agent de lutte biologique contreMyzus persicae Sulzer. Les larves du 2ème stade élevées en laboratoire de ce chrysope à raison de 6 par plante sont les plus actives et réduisent la population de pucerons de 78% en 2 semaines. Les sécrétions des trichomes des feuilles de tabac n'ont pas d'effet sur les larves ni sur leurs mouvements. Les larves se déplacent librement et se nourissent bien; des nymphes ont été trouvées sur tous les plants, ce qui indique que le prédateur est valable contreM. persicae et bien adapté à la culture de tabac.

     

Production and characteristics of inhibitory factor of raw starch digestion from Aspergillus niger

Summary The glucoamylase preparation of Aspergillus niger 19 inhibited the raw starch digestion by it at high enzyme concentration. The inhibitory factor (IF) was isolated from the glucoamylase preparation by heat treatment and purified by DEAE-Sephadex A-25 column chromatography, an initial Sephadex G-50 gel filtration followed by SP-Sephadex C-25 column chromatography (twice) and then second Sephadex G-50 gel filtration. The IF thus purified was homogenous in polyacrylamide gel electrophories. The inhibitory activity of IF increased with the increasing IF concentration but decreased with an increasing quantity of raw starch or enzyme concentration. The IF had no effect on the hydrolysis of boiled soluble starch. It was completely adsorbed onto raw starch. The IF had a molecular weight of about 10,500. It was abundant in hydroxy amino acids such as threonine and serine. Xylose, mannose, glucose, galactose, and galacturonic acid were present in it.     

Immunochemical evidence for the presence of 5mC, 6mA and 7mG in human, Drosophila and mealybug DNA

We have reported that production and characterization of antibodies highly specific to 5-methyl-cytosine (5mC) and the development of a sensitive immunochemical method for the detection of 5mC in DNA [FEBS Lett. (1982) 150, 469]. Extension of this method to two other modified bases, 6-methyladenine (6mA) and 7-methylguanine (7mG), is reported here. By use of this immunochemical approach, we are able to detect 5mC, 6mA and 7mG in human and Drosophila DNA and confirm their presence in the DNA of two mealybug species.     

Feasibility of Outpatient Self-Administration of Parenteral Antibiotics

It is customary to treat patients with infective endocarditis or osteomyelitis for a prolonged period in hospital with parenteral antibiotics. It was felt that it might be feasible to allow parenteral administration of antibiotics by the patients themselves at home. Results in 13 patients who administered antibiotics parenterally themselves (experimental group) were compared with those in 7 patients (control group) treated entirely within the hospital. Antibiotic-related complications were similar in both groups. There was no instance of infection of the intravenous cannula in either group. The average daily cost of antibiotic therapy decreased from $243.22 for inpatients to $69.35 for outpatients. The average cost of illness was $6,357.22 in the experimental group and $10,022.23 in the control group. If patients are carefully selected and well educated, the outpatient self-administration of antibiotics parenterally is both economical and safe.     

Separation of poly(A) RNA's synthesized by soybean embryos

A stepwise elution procedure in which the MAK column with 0.85M buffered NaCl is held in a stationary phase (2–3 psi,35?C, 1 hr) permits the separation of two tenaciously boundRNA fractions. Poly(A) RNA's elute with the ribosomal and tenaciouslybound RNA's. MAK fractions contain different percent of RNA'sthat bind to poly(dT) cellulose and have messenger RNA activity.The results suggest that the soybean embryonic tissue synthesized,at least, four poly(A) RNA's. (Received August 17, 1976; )     

Release of Esterase Following Germination of Lettuce Seed (Lactuca sativa L.)

Light-insensitive lettuce seeds, Lactuca sativa L. cv. Great Lakes, release esterases for a period following radicle protrusion. Very little or no enzymes are released prior to 24 hours or after 48 hours of germination. As compared to intact seeds, half-seeds readily release esterases and the release is not affected by far red irradiation. Bulk of the released esterases are derived from the endosperm tissue and presumably exists in the intact seed as a component of the extraembryonic fluid.     

Influence of 4′-6-diamidino-2-phenylindole on the secondary structure and template activities of DNA and polydeoxynucleotides

Summary The interaction between 4-6-Diamidino-2-Phenylindole-hydrochloride (DAPI) and a variety of DNAs and synthetic polydeoxynucleotides was investigated in order to delineate the nucleic acid structural features necessary for binding. The spectra of DAPI-DNA complexes, measured at various DAPI-DNA molar ratios (r), are hypochromic relative to DNA in the region of its maximum absorption. All the curves pass through an isosbestic point at 268 nm. A new maxima appears in the region of 380–392 nm for DAPI-DNA complexes. The magnitude of the peaks in the region are directly proportional to the amount of drug present in the complex.Studies with various DNA types and synthetic polydeoxynucleotides indicate that the drug preferentially binds to dAT-rich regions of DNA. This was also confirmed by enzymatic studies. The inhibition of template action by DAPI in a purified DNA-polymerase reaction was dependent on the dAT-content of the template. The implication of these data to explain a selective binding of DAPI to mitochondrial DNA have been discussed.