Replication-competent picornaviruses with complete genomic RNA 3' noncoding region deletions.

The genomic RNA 3' noncoding region is believed to be a major cis-acting molecular genetic determinant for regulating picornavirus negative-strand RNA synthesis by promoting replication complex recognition. We report the replication of two picornavirus RNAs harboring complete deletions of the genomic RNA 3' noncoding regions. Our results suggest that while specific 3'-terminal RNA sequences and/or secondary structures may have evolved to promote or regulate negative-strand RNA synthesis, the basic mechanism of replication initiation is not strictly template specific and may rely primarily upon the proximity of newly translated viral replication proteins to the 3' terminus of template RNAs within tight membranous replication complexes.     

Mapping the diabetes polygene Idd3 on mouse Chromosome 3 by use of novel congenic strains

Development of novel congenic mouse strains has allowed us to better define the location of the diabetogenic locus, Idd3, on Chromosome (Chr) 3. Congenic strains were identified by use of published and newly developed microsatellite markers, their genomes fingerprinted by a rapid, fluorescence-based approach, and their susceptibility to type 1 diabetes evaluated. The maximum interval containing Idd3 is now approximately 4 cM.     

Studies on growth hormone secretion IX. Prostaglandins do not act like ionophores

To gain further insight on the mechanism of GH secretion in general and on the stimulation of this process by prostaglandins in particular, we compared the effects of PGE₁ and PGE₂ on hormone release and cyclic nucleotide levels with those of the ionophores A23187 and X537A under a variety of experimental conditions. All these substances (in the presence but not in the absence of calcium) enhanced GH release in incubated rat anterior pituitaries , prostaglandins being considerably more potent than ionophores. However, while PGE₂ caused a dose-dependent rise in pituitary cyclic AMP levels (from doubling at 10⁻⁷ M to a two-hundred fold increase at 10⁻⁵ M), the ionophores had no effect on the concentrations of this nucleotide. Neither PGE₂ nor the ionophores had any measurable effect on cyclic GMP levels. Exposure of tissues to ionophores for 60 minutes rendered them refractory to subsequent stimulation by PGE₁ or to ionophores themselves, whereas preincubation with PGE₁ did not diminish GH responses during a second incubation period. On the other hand, 60-minute preincubation of hemipituitaries in the presence of ionophores (10⁻⁵ M) did not suppress subsequent PGE₁-promoted cyclic AMP accumulation. Metabolic blockers inhibited PGE₂ and A23187-promoted GH-release but failed to suppress GH-response to X537A. Verapamil partially inhibited PGE₂ but not ionophore induced GH secretion. Ionophores particularly X537A, accelerated 45Ca efflux while PGE₁ did not influence this. Electronmicroscopy revealed extensive vacuolization localized chiefly at the Golgi apparatus when tissues were incubated with X537A. PGE₁ and A23187 had no such morphological effect. It is concluded that prostaglandins E and ionophores promote GH secretion by different mechanisms.     

Diagnosis of encephalitozoonosis in experimentally infected rabbits by intradermal and immunofluorescence tests.

The indirect immunofluorescence antibody test was performed on serial blood samples from eight young New Zealand White rabbits with experimental encephalitozoonosis. The test showed seroconversion in six of the eight infected rabbits by the 8th day after inoculation and in all rabbits by the 15th day. Antibody titers reached a peak by about the 36th day after inoculation and remained significantly elevated until the termination of the experiment at 84 days after inoculation. None of four sham-inoculated rabbits showed an immunofluorescence response by the 60th day after inoculation. Immunofluorescence and intradermal test responses were compared before infection and at the 60th day after inoculation in a total of 32 experimentally infected rabbits. Both tests were equally effective (100%) in detecting infected animals. Six of eight (first group) and 22 of 24 (second group) experimentally infected rabbits were confirmed histologically to have lesions compatible with encephalitozoonosis. No cross reactions were observed between Encephalitozoon cuniculi and Toxoplasma gondii, Eimeria perforans, or Eimeria stiedai by intradermal test or immunofluorescence test.     

Cycling of organic and inorganic sulphur in a chestnut oak forest

Summary Sulfur (S) cycling in a chestnut oak forest on Walker Branch Watershed, Tennessee, was dominated by geochemical processes involving sulfate. Even though available SO ₄ ^2- was present far in excess of forest nutritional requirements, the ecosystem as a whole accumulated 60% of incoming SO₄–S. Most (90%) of this accumulation occurred by SO ₄ ^2- adsorption in sesquioxide-rich subsurface soils, with a relatively minor amount accumulating and cycling as SO ₄ ^2- within vegetative components. Organic sulfates are thought to constitute a large proportion of total S in surface soils, also, and to provide a pool of readily mineralized available S within the ecosystem.Research sponsored by Division of Biomedical and Environmental Research, U.S. Department of Energy, under contract W-7405-eng-26 with Union Carbide Corporation. Soil ester sulfate work sponsored by contract RP-1813-1 with the Electric Power Research Institute. Publication No. 1990, Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37830     

The nitrogen cycle in a ‘Terra Firme’ rainforest on oxisol in the Amazon territory of Venezuela

Standing stocks and fluxes of nitrogen, including nitrogen fixation and denitrification, were measured in a tropical rainforest on Oxisol in the Amazon Territory of Venezuela. The standing stock of nitrogen was comparable to that of temperate forests, but was higher than that in an adjacent forest on Spodosol. Fluxes were higher than in forests in the temperate zone, but lower than in another tropical forest on more fertile soil. Even though nitrogen was abundant, this does not mean that nitrogen could not be limiting to agriculture if the forest is cleared and the land cultivated. The nitrogen fixing and nitrogen conserving mechanisms are dependent upon the structure of the undisturbed forest, and destruction of the forest would eventually decrease the input of nitrogen to the soil.     

Improved Ultrastructural Detail in Tissues Fixed with Potassium Permanganate

An improved method has been developed for fixation with potassium permanganate. Although this is one of the methods widely used to preserve the dense cores of adrenergic storage vesicles, fixation of other tissue components is usually poor. The main differences from previously reported methods using potassium permanganate are the use of a physiological saline as the vehicle for all solutions, and, following this, very rapid dehydration before infiltration with plastic. Cellular and intercellular details of tissue ultrastructure may, in general, be evaluated as satisfactorily as with conventional fixatives, with the exception of certain protein elements associated with ribosome, microtubule, and myofilament organization. Nerve endings with agranular or clear vesicles may be distinguished from adrenergic endings since the dense cores of the vesicles of the latter are preserved by this method.     

Biliary excretion of synthetic sex steroids in heterozygous and homozygous Gunn rats.

87.9% of a given dose of [³H]Norethisterone ([³H]N) and 76.7% of [³H]Ethinyloestradiol ([³H]EE₂) were excreted in the bile of male heterozygous Gunn rats in 2 hours Similarly, 86.9% of a given dose [³H]N and 84.0% of [³H]EE₂ were excreted in the bile of male homozygous Gunn rats in 2 hours. In both heterozygous and homozygous rats glucuronide conjugates were present. Despite the lesion in UDP-glucuronyltransferase, the homozygous rats is able to conjugate the synthetic steroids apparently normally.