Eleven structurally similar ketolide antibiotics were tested at a concentration of 1 μg/ml for their relative inhibitory effects on growth and ribosome activities in Staphylococcus aureus cells. Ten of the compounds examined had an inhibitory effect on protein synthesis at this concentration and eight of the 11 compounds were also effective inhibitors of the formation of the 50S ribosomal subunit. All of the drugs tested inhibited protein synthesis to a greater extent than they affected 50S subunit formation. The decline in growth rate and cell number was proportional to the effect on ribosome formation and function. The growth of an ermC erythromycin-resistant strain of S. aureus was also significantly inhibited by nine ketolide compounds, suggesting that they were not inducers of methylase gene expression. These inhibitory activities can be related to structural differences between these ketolide antibiotics.

Nine structurally similar macrolide antibiotics were tested at a concentration of 0.5 μg/ml for their relative inhibitory effects on ribosome functions in Staphylococcus aureus cells. Eight of the compounds examined inhibited protein synthesis at this concentration. Seven of the nine compounds were also effective in blocking formation of the 50S ribosomal subunit. Roxithromycin and 14-hydroxy clarithromycin inhibited protein synthesis to a greater extent than they affected 50S subunit formation. Conversely, the compound 11,12-carbonate-3 deoxy-clarithromycin affected 50S assembly more than translation. Only clarithromycin had any effect on 30S ribosomal subunit assembly. The decline in growth rate and cell number was proportional to the effect on ribosome formation or function by each compound. These inhibitory activities can be related to structural differences between these macrolide antibiotics.

The ID50 values for azithromycin and clarithromycin inhibition of translation and of 50S assembly in Staphylococcus aureus cells have been measured. For clarithromycin, 50% inhibition of growth occurred at 0.075 μg/ml, and the effects on translation and 50S formation were equivalent at 0.15 μg/ml. The inhibition of these processes by azithromycin was less effective, with an ID50 of 2.5 μg/ml for growth and 5 μg/ml for inhibition of translation and 50S formation. The additive effects of each of these drugs on translation and 50S formation account quantitatively for their observed influence on cellular growth rates. In macrolide-treated cells, there was also a direct relationship between the loss of ribosomal RNA from the 50S subunit and its accumulation as oligoribonucleotides. These results are compared with the previously described effects of erythromycin on these same processes.

Erythromycin and other macrolide antibiotics have been examined for their effects on ribosome assembly in growing Escherichia coli cells. Formation of the 50S ribosomal subunit was specifically inhibited by erythromycin and azithromycin. Other related compounds tested, including oleandomycin, clarithromycin, spiramycin, and virginiamycin M1, did not influence assembly. Erythromycin did not promote the breakdown of ribosomes formed in the absence of the drug. Two erythromycin-resistant mutants with alterations in ribosomal proteins L4 and L22 were also examined for an effect on assembly. Subunit assembly was affected in the mutant containing the L22 alteration only at erythromycin concentrations fourfold greater than those needed to stop assembly in wild-type cells. Ribosomal subunit assembly was only marginally affected at the highest drug concentration tested in the cells that contained the altered L4 protein. These novel results indicate that erythromycin has two effects on translation, preventing elongation of the polypeptide chain and also inhibiting the formation of the large ribosomal subunit.

The binding of the aminocyclitol antibiotic spectinomycin to 70S ribosomes and to 30S subunits from Escherichia coli has been investigated. The association was influenced by the presence of messenger RNA. The Kd for [3H]-4 OH-spectinomycin binding to 70S ribosomes was 2×10-7 M without mRNA (polyinosinic acid), and 1×10-6 M with polyinosinic acid. Dissociation of the antibiotic from the ribosomes was significantly affected by the presence of a bound messenger RNA, which reduced the rate of dissociation by a factor of 5.7. The presence of mRNA did not influence the association of spectinomycin with the 30S subunit. The dissociation rate from the small subunit was comparable to the rate of dissociation from the 70S ribosome and was not affected by the presence of mRNA. © 1990 Springer-Verlag New York Inc.

Trypsin, when added to a bioassay for tumoricidal macrophages, produced killing of tumor cells. Trypsin cleaved fetal calf serum proteins to produce a protein fragment that activated macrophages to lyse tumor cells. Diisopropyl fluorophosphate-inhibited trypsin did not produce tumoricidal macrophages either by direct action on the macrophage or by action on serum proteins. The macrophage activation factors produced from serum proteins were fractionated into molecular weight ranges of 150,000, 68,000, and 30,000-5000. The effects of neutral proteinases and proteinase inhibitors on the ability of macrophages to lyse tumor cells is discussed.

Osmotically induced alterations in the ultrastructure of the apical crypts of chloride cells and changes in chloride efflux were studied in striped bass (Morone saxatilis). Striped bass were divided into three groups: fish adapted to freshwater, fish transferred directly from freshwater to 100% seawater (3% salt, w/v) for 24 hr or less, and fish adapted to 100% seawater for 7 days or more. Transmission electron microscopy studies revealed multicellular complexes of cells in both freshwater‐ and seawater‐adapted fish. Cytoplasmic indigitations between cells in the complex were more numerous in seawateradapted bass. Scanning electron microscopy studies showed that the apical extensions in freshwater fish were uniform in size. Changes in ultrastructure and chloride efflux were observed within 3 hr after transfer to seawater. Initially the apical extensions of chloride cells become longer, more prominent, and branched. After 7 days in seawater some of the apical crypts develop into a deeper “pit” structure, while others remain like those of freshwater fish. An increase in the number of apical crypts is measured by 14 days after transfer. Chloride efflux increases to five times freshwater values after 24 hr and 17 times freshwater values after 7 days in seawater. Mitochondrial density is not significantly different between freshwater and seawater fish (7 or more days). The response of chloride cell apical crypts is not an all‐or‐none phenomenon as observed in other species. Striped bass are able to increase chloride efflux when osmotically stressed with little ultrastructural alteration.

The surface ultrastructure of the gill arches of the killifish, Fundulus heteroclitus, adapted to seawater or freshwater, was found to be similar to that reported for other euryhaline teleosts. Two rows of gill filaments (about 42 filaments per row) extended posterolaterally, and two rows of gill rakers (about 10 rakers per row) extended anteromedially from each arch. Leaf‐like respiratory lamellae protruded along both sides of each filament, from its base to its apex. The distributions, sizes, and numbers of various surface cells and structures were also determined. All surfaces were covered by a mosaic of pavement cells, which measured about 7 × 4 μm and exhibited concentrically arranged surface ridges. Taste buds were especially prominent on the rakers and the pharyngeal surfaces of the first and second gill arches, but were often replaced by horny spines on the third and fourth gill arches. Apical crypts of chloride cells occurred mostly on the surfaces of the gill filaments adjacent to the afferent artery of the filament. In seawater adapted killifish, crypts resembled narrow, deep holes along the borders of adjacent pavement cells, had openings of about 2 μm2, and occurred at a frequency of about 1 per 70 μ2 of surface area. In freshwater fish, the crypts usually had larger openings (about 10 μ2), occurred less frequently (1 per 123 μ2), and exhibited many cellular projections in their interiors. Changes in crypt morphology may be related to the ion transport function of chloride cells.

This paper presents a study which utilizes routine transmission (TEM) and scanning (SEM) electron microscopy, and vascular corrosion casting (VCC) to describe the normal microvasculature of the rabbit bladder. A clear 3-D view of the microvasculature of the normal rabbit bladder is provided by the methods used in this study.

Rats were given bilateral aspiration lesions of the hippocampus. Some of these rats then received bilateral transplants of fetal hippocampal or dorsal ventricular ridge tissue that was dissected from embryonic rat brains at 16 or 17 days of gestation. The remaining rats with hippocampal lesions did not receive fetal brain transplants. Rats with neocortical aspiration lesions, but without transplants, and rats without brain damage were also included in the study. All of the rats were trained to find a submerged platform in a Morris water maze. Rats with the fetal brain transplants were more impaired in some measures of maze learning than were rats with hippocampal lesions only. The results indicate that transplants of fetal brain tissue are not always associated with recovery of behavioral function after brain damage and may even increase a lesion-induced behavioral impairment in tasks that require complex cognitive functioning.