Curvature of mouse satellite DNA and condensation of heterochromatin

Cloned, sequenced mouse satellite DNA exhibits properties characteristic of molecules that possess a stable curvature. Circularly permuted fragments containing the region predicted to bend were used to map the curvature relative to DNA sequence. The altered mobility of these fragments in polyacrylamide gels is reversed when gels are run in the presence of distamycin A, a drug that binds preferentially to AT-rich DNA. Treatment of living mouse cells with this drug dramatically reduces the condensation of centromeric heterochromatin, the exclusive location of satellite sequences. In situ hybridization of satellite probes to extended chromosomes at the electron microscope level shows that satellite does not comprise a single block but is distributed throughout the centromere region. Based on these experiments, we hypothesize that the structure of mouse satellite DNA is an important feature of centromeric heterochromatin condensation.     

Necator americanus: temperature, pH, light, and larval development, longevity, and desiccation tolerance

The effect of incubation temperature and pH on the hatch rate of eggs of Necator americanus, and the desiccation tolerance of the resulting infective stage-3 larvae were investigated in the laboratory under controlled conditions. Hatching did not occur below 15 C and above 35 C. A 21% hatch rate was obtained at 15 C while a 10.6% hatch rate was obtained at 35 C. The highest hatch rate (93.7%) was obtained at 30 C. The optimum pH for hatching was 6.0, but the larvae did not reach the infective stage. Incubation temperature of the eggs affected the longevity and desiccation tolerance of resultant infective larvae. Larvae hatched at 30 C and maintained at 26 C under bright fluorescent light had a 50% survival time (S50) of 4 days. In the dark or shade, the S50 for larvae raised at 30 C was 5 weeks, while that of larvae hatched at 20 C was 7 weeks. Incubation temperature also affected the desiccation tolerance of larvae. Larvae developed at 20 C were more resistant to desiccation at various relative humidity values than larvae hatched at 30 C.     

Analysis of S-methylmethionine and S-adenosylmethionine in plant tissue by a dansylation, dual-isotope method

A method is presented for determining the levels of S-methylmethionine (MeMet) and S-adenosylmethionine (AdoMet) in the same plant tissue sample, utilizing readily available equipment. The bottom limit of sensitivity, ca. 100 pmol, can be lowered if required. A trichloracetic acid homogenate of the tissue is supplemented with [carboxyl-14C]MeMet and [carboxyl-14C]AdoMet. After separation of MeMet and AdoMet from each other and from endogenous homoserine on a phosphocellulose column, the two fractions are heat treated at appropriate pH values to liberate [14C]homoserine. Quantitation is via the 3H/14C ratio of [3H]dansyl-[14C]homoserine isolated by thin-layer chromatography. The method is validated with pea cotyledon, corn root, and cauliflower leaf.     

Methyl iodide, a potent inducer of the adaptive response without appreciable mutagenicity in E. coli

Methyl iodide (MeI), a very weak mutagen, induced the adaptive response in E. coli to a similar extent to those induced by potently mutagenic methylating agents. MeI potentiated the mutagenicity of a methylating mutagen, N-methyl-N-nitrosourea, by its co-treatment. These results might give indication that MeI directly methylates O6-methylguanine-DNA methyltransferase resulting in induction of the adaptive response and depletion of the repair capacity of enzyme.     

Autochthonous fungi are central components in microbial community structure in raw fermented sausages

Raw meat sausage represents a unique ecological niche rich in nutrients for microbial consumption, making it particularly vulnerable to microbial spoilage. Starter cultures are applied to improve product stability and safety as well as flavour characteristics. However, the influence of starter cultures on microbial community assembly and succession throughout the fermentation process is largely unknown. In particular the effect on the fungal community has not yet been explored. We evaluate the microbiological status of four different raw meat sausages using high-throughput 16S rRNA gene and ITS2 gene sequencing. The objective was to study temporal changes of microbial composition during the fermentation process and to identify potential keystone species that play an important role within the microbial community. Our results suggest that fungi assigned to the species Debaryomyces hansenii and Alternaria alternata play a key role in microbial community dynamics during fermentation. In addition, bacteria related to the starter culture Lactobacillus sakei and the spoilage-associated genera Acinetobacter, Pseudomonas and Psychrobacter are central components of the microbial ecosystem in raw fermented sausages. Elucidating the exact role and interactions of these microorganisms has the potential to have direct impacts on the quality and safety of fermented foods.     

Mice: postpartum aggression is elevated following prenatal progesterone exposure

Pregnant Rockland-Swiss (R-S) mice were injected with sesame oil or 250 or 500 micrograms of progesterone (P) on Days 12 through 16 of gestation and the postpartum aggressive behavior of their female offspring was examined in adulthood. Both doses of P significantly increased the intensity of aggression (number of attacks) exhibited by the female offspring toward an adult male intruder. The low dose of P also produced significant increases in relative anogenital distance. These effects were seen in the absence of any effects on body weight at birth or in adulthood, or on reproductive performance. The findings support previous research, in both animals and humans, showing that prenatal brain differentiation and subsequent behavior are masculinized by prenatal exposure to progesterone.     

Comparison of the inhibitory effects of mexiletine and lidocaine on the calcium current of single ventricular cells

Effects of mexiletine and lidocaine on inward calcium current (ICa) of single ventricular myocytes from guinea pigs were studied using tight seal whole cell clamp method. Mexiletine at the concentrations of 10, 30 and 100 microM decreased ICa by 23.0, 28.9 and 55.4%, respectively, while lidocaine decreased it by 8.9, 16.8 and 25.2%. At all concentrations tested, a potency for ICa inhibition in mexiletine was significantly greater than that in lidocaine (p less than 0.05). The results suggest that mexiletine has, at therapeutic concentrations, a considerable blocking action on the Ca channels other than well-known action on the Na channels.     

Closed channel-open channel equilibrium of the sodium channel of nerve. Simple models of macromolecular equilibria

The consistency of an electrodiffusion kinetics to describe the time-dependent opening of sodium channels of nerve suggests that motions over relatively long distances (on the atomic scale) are involved in the equilibrium as well. As a result, it is expected that a relatively large fraction of possible macromolecular conformations are unreactive. An equilibrium constant between locally reactive forms and the unreactive conformations is introduced. The consequences of this formalism is investigated in a square well potential, a harmonic potential, and a system consisting of two harmonic potentials with different spatial extents. The limits of knowledge from Nernstian behavior are shown. As an alternative to the Nernstian analysis, the experimental data of the sodium channel's quasi-equilibrium - the probability of the channel's being open as a function of voltage - can be described as resulting from motion caused by an electric field on a charge which is confined by a harmonic potential. A force constant is found from this analysis. (Such Hookian force constants cannot be found from spectroscopic experiments in condensed systems where the large-displacement vibrations are overdamped and, hence, spectroscopically unobservable). From the force constant, an approximate value of the Young's modulus can be calculated. The modulus' value falls in the range for rubber. As for rubbers, the restoring force is, then, expected to be mostly entropic rather than enthalpic in origin. Using the appropriate theory for linear chains of rubber and the Young's modulus, the approximate length of the chain causing the rubber-like force is calculated. The result is found to be near the length suggested for the hydrophilic chains that connect transmembrane sections of the sodium channel.     

Antibacterial hemolymph proteins of Manduca sexta

Exclusion column fractionated immune hemolymph of the M. sexta larva contains five peaks of anti-E. coli activity with molecular weights of greater than 140 kD and approximately 91, 54, 14 and 4 kD, plus one peak of lysozyme activity with a molecular weight of 17 kD. Purification of the 54 kD peak showed that this peak consists of the previously described M18 proteins which have monomeric weights of approximately 20 kD and had antibacterial activity against certain gram negative bacteria. Approximately 80% of the total hemolymph antibacterial activity was detected in the 14 and 4 kD peaks. These proteins, which kill both gram negative and gram positive bacteria, appeared to be directly analogous to the cecropins of H. cecropia. The greater than 140 and 91 kD peaks constituted only a minor part of the total antibacterial activity.     

Identification of ten additional nucleotides in the primary structure of yeast 18S rRNA

The ten-nucleotide-long sequence have been omitted while sequencing the 18S rRNA gene from yeast Saccharomyces cerevisiae [Rubtsov et al., Nucl. Acids Res. 8 (1980) 5779-5794]. This GAAGAUGAUC sequence and some other minor corrections are reintroduced into the yeast 18S rRNA primary structure.