Molecular recognition of anions by synthetic receptors

Over the past year, several significant developments have been made in the field of anion binding. The fundamental principles of molecular recognition are increasingly being better understood, and of particular interest are reports of several synthetic anion receptors able to perform their task in complex natural media. Additionally, macroscopic devices such as anion specific electrodes and membranes based on a molecular recognition approach are now being made.     

Repair mechanisms involved in muscle regeneration following partial excision of the rat gastrocnemius muscle

The sequential cytological events of the regeneration process, after partial excision of the gastrocnemius muscle in the rat, were followed by light and electron microscopy. During the first 2 days after injury leukocytes and macrophages infiltrate into the traumatized area. Myogenic regeneration is then characterized by mainly two repair mechanisms. Mononucleated cells, that populate the excised area, most probably fuse together to give rise to newly formed multinucleated myotubes that further develop to striated myofibers. Another mechanism involves the repair of injured muscle fibers by the possible fusion of mononucleated cells with their necrotic cut ends. Consequently, by addition of nuclei and new muscular material, sarcoplasmic outgrowths from the injured fibers are formed. It is concluded that mainly two repair mechanisms are involved in the regeneration process following partial excision of a muscle: addition of new muscle fibers in a process similar to that of embryonic myogenesis and also meristic growth from the injured fibers.     

Stream drift,size-specific predation,and the evolution of ovum size in an amphibian

Summary A stream-breeding race of small-mouthed salamanders (Ambystoma texanum) in central Kentucky produces ova that are twice as large as those of a pond-breeding race found nearby. Embryos of stream-breeders also hatch at a more advanced developmental stage than those of pond-breeders. Morphological evidence indicates that stream-breeders were derived from pond-breeding stock. Assuming that differences between stream and pond-breeders reflect evolutionary change, and that the ancestral pond stock that invaded streams was similar to extant pond-breeders, we examined three hypotheses that might explain changes in ovum size and stage at hatching following the invasion of streams. (1) Larger ovum size evolved indirectly as a consequence of selection for rapid development which minimizes mortality risk from stream drying. (2) Increased ovum (hatchling) size and stage at hatching of stream-breeders are adaptations to resist stream current. (3) Increased ovum (hatchling) size and stage at hatching are adaptations to reduce predation on hatchlings from stream invertebrates. The results of field and laboratory studies only support hypotheses (2) and (3). Hatchlings that were relatively large or at a more advanced developmental stage had slower drift rates and were less vulnerable to predation by Phagocata gracilis, a flatworm abundant in streams in central Kentucky. Developmental and growth parameters were not correlated significantly with ovum size in populations of either geographic race. Differences in degree of parental care among races also cannot explain variation in ovum size since both races abandon their eggs immediately after oviposition.     

A physical domain of herpes simplex virus ICP8 is expressed and active in Escherichia coli.

In this report, we describe a series of procedures to assay the function of fusion genes in Escherichia coli and the specific application to the carboxy-terminal third of the herpes simplex virus type 1 (HSV-1) DNA-binding protein ICP8. E. coli cells containing the cloned HSV-1 BamHI G fragment with the HSV-1 BamHI-G-V site, map unit 0.388, nearest the tet promoter in pBR322 synthesized an active product containing a portion of ICP8. The new product induced phenotypic alterations in recipient hosts that were measurable and stable yet limited to the stability of the plasmid. The corresponding cloned DNA from the characterized HSV-1 DNA-binding protein mutant tsHA1 exhibited a predictable temperature-sensitive phenotype. Screening procedures based on the loss of induction of the parental plasmid-induced phenotype in E. coli cells allowed us to select additional mutations. One of these, which conferred a phenotype different from that of tsHA1, was transferred to the viral genome by marker transfer techniques. We suggest that any mutant could be isolated in any sequence, provided that the wild-type coding sequences induce alterations in E. coli cells. The observed alterations should have relevance in determining the mode of action of the protein in its normal environment.     

Antagonism by theophylline of respiratory inhibition induced by adenosine

The effects on respiration of an analogue of adenosine, L-2-N6-(phenylisopropyl)adenosine (PIA), and of the methylxanthine, theophylline, were determined in 19 vagotomized glomectomized cats whose end-tidal PCO2 was kept constant by means of a servo-controlled ventilator. Integrated phrenic nerve activity was used to represent respiratory output. Our results show that PIA, whether given systemically or into the third cerebral ventricle, depressed respiration. Systemically administered theophylline stimulated respiration. Theophylline given intravenously, or into the third ventricle not only reversed the depressive effects of previously administered PIA but caused further increases of respiration above the control level. Prior systemic administration of theophylline blocked both respiratory and hypotensive effects of subsequently administered PIA. Effects of either agent on medullary extracellular fluid pH did not explain the results. We conclude that the adenosine analogue PIA, acts to inhibit neurons in the brain that are involved in the control of respiration and that its effects are blocked by theophylline. We suggest that adenosine acts as a tonic modulator of respiration and that theophylline stimulates breathing by competitive antagonism of adenosine at neuronal receptor sites.     

Thiostrepton resistance mutations in the gene for 23S ribosomal RNA of halobacteria

Mutants of Halobacterium (H.) halobium and H. cutirubrum were isolated which are resistant to the 70S ribosome inhibitor thiostrepton. Using primer extension analysis, resistance was shown to correlate with base changes at position 1159, which corresponds to position 1067 of the E. coli 23S rRNA. In four mutants, A1159 was replaced by U, in one mutant by G. The results show that not only methylation (Cundliffe & Thompson (1979) Nature 278, 859-861) of A1067 (E. coli nomenclature), but also base changes at this position cause high-level resistance to thiostrepton.     

Circumsporozoite protein of Plasmodium berghei: gene cloning and identification of the immunodominant epitopes.

The gene encoding the circumsporozoite (CS) protein of the rodent malaria parasite Plasmodium berghei was cloned and characterized. A cDNA library made from P. berghei sporozoite RNA was screened with a monoclonal antibody for expression of CS protein epitopes. The resulting cDNA clone was used to isolate the CS protein gene from a lambda library containing parasite blood-stage DNA. The CS protein gene contains a central region encoding two types of tandemly repeated amino acid units, flanked by nonrepeated regions encoding amino- and carboxy-terminal signal and anchorlike sequences, respectively. One of the central repeated amino acid unit types contains the immunodominant epitopes.