A NEW CHONDRODYSTROPHIC MUTANT IN MICE : Electron Microscopy of Normal and Abnormal Chondrogenesis

The occurrence of a new mutation affecting cartilage and bone in mice is reported. The gene is lethal, shows autosomal recessive inheritance, and has high penetrance. It is not allelic to shorthead and probably not to phocomelia or achondroplasia. It results in a foreshortened face, cleft palate, defective trachea, and shortened long bones with flared metaphyses. Chondrocytes of epiphyseal cartilage from the mutant are not aligned in columns, and there is a decrease in the usual staining of the cartilage matrix. Electron microscope observations show large, wide collagen fibrils with "native" banding in the matrix of mutant cartilage, which are not present in normal cartilage. Possible explanations for the expression of this genetic disorder of cartilage development are put forward.     

Cumulation of active metabolites of levo-alpha-acetylmethadol in the rat fetus and neonate

Levo-alpha-acetylmethadol (LAAM, 0.2 or 2.0 mg/kg/day) was orally administered to female Sprague-Dawley rats for one month prior to and throughout pregnancy. The rats were killed on the 18th day of gestation along with a group of 18-day pregnant females given a single oral 2.0 mg/kg dose of LAAM 24 hours earlier. Although cumulation of LAAM or its active metabolites was not seen in plasma or brain of pregnant rats given drug chronically, significant cumulation was observed in whole fetus and in fetal brain. In addition, a 2–3 fold elevation in the concentrations, and an even greater elevation of total content, was noted in the newborn pup. These data suggest that opiate intoxication soon after birth may be a factor responsible for the increased morbidity and mortality of rat pups prenatally exposed to LAAM.     

The mutagenicity of heterocyclic N-nitrosamines for Salmonella typhimurium

14 carcinogenic and noncarcinogenic heterocyclic N-nitrosamines were evaluated for mutagenicity to Salmonella typhimurium TA-1535, which responds to mutagens inducing base-pair substitutions. Both suspension and plate tests were used, with mouse and rat liver in vitro metabolic activation systems.All carcinogenic nitrosamines showed a positive response in at least one test system, as did the noncarcinogens. In general, the mutagenic responses obtained with mouse liver were equal to, or greater than, the responses obtained with rat liver in both the suspension and plate tests. Although it is difficult to make quantitative comparisons between plate and suspension tests, both systems appeared to be responsive to the same dose ranges for the individual nitrosamines.     

Factors affecting the contribution by epiphytic algae to the primary productivity of an oligotrophic freshwater lake

A diatom-dominated population of epiphytic algae was studied in an oligotrophic lake to determine the factors which limit epiphyte growth and to measure their contribution to primary productivity. Algae were collected from plants growing at four sites in Lake George, N.Y., during the spring, summer, and fall of 1974. Samples were taken from 3 m, corresponding to the depth at which macrophytes were most productive. Algae exhibited an optimum temperature for H¹⁴CO₃^- uptake at 30 C, although the summer littoral lake temperature ranged from 18 to 25 C. Light saturation occurred at an intensity of 8,608 lux, approximating the environmental intensity at the depth from which algae were taken. Epiphytes exhibited their maximum photosynthetic capacity of 0.6 mg of carbon fixed/m² of macrophyte surface area per h in the early afternoon in mid-August. They assimilated approximately 5% as much inorganic carbon as the macrophytes from which they were taken. Epiphyte population densities followed the seasonal growth patterns of the macrophytes, with maximal leaf colonization remaining essentially constant relative to the leaf position on the plant. There was little change in density between sampling sites at any given time. Productivities of epiphytes from bottom leaves were 10-fold greater than those of epiphytes from top leaves. Addition of PO₄^-3, NO₃^-, NH₃, Si, and SO₄^-2 had no stimulatory effect on photosynthesis. Addition of HCO₃^- stimulated photosynthesis greater than 30%, suggesting that carbon may be a limiting nutrient for epiphytic algae in Lake George.     

Distribution of sister chromatid exchanges in the euchromatin and heterochromatin of the Indian muntjac

The frequency of sister chromatid exchanges (SCEs) was determined for the chromosomes (except Y2) of the Indian muntjac stained by the fluorescence plus Giemsa (FPG) or harlequin chromosome technique. The relative DNA content of each of the chromosomes was also measured by scanning cytophotometry. After growth in bromodeoxyuridine (BrdU) for two DNA replication cycles. SCEs were distributed according to the Poisson formula in each of the chromosomes. The frequency of SCE in each of the chromosomes was directly proportional to DNA content. A more detailed analysis of SCEs was performed for the three morphologically distinguishable regions of the X-autosome composite chromosome. The SCE frequency in the euchromatic long arm and short arm were proportional to the amount of DNA. In contrast, the constitutive heterochromatin in the neck of this chromosome contained far fewer SCEs than expected on the basis of the amount of DNA in this region. A high frequency of SCE, however, was observed at the point junctions between the euchromatin and heterochromatin.     

An electrophysiological study of the membrane properties of the immature and mature oocyte of the Batstar,Patiria miniata

Immature oocyte membrane properties of a starfish, Patiria miniata, were investigated by microelectrode techniques. The resting membrane potential in artificial seawater (ASW) was ?78.5 ± 6.7 mV (n = 61, inside negative). This was mainly accounted for by a selective permeability to potassium ions. Potassium ion-selective microelectrodes were used to measure intracellular K⁺ ion activity, which was 350 mM. The sodium to potassium permeability ratio was 0.02 ± 0.01 (n = 4). The current-voltage relation was nonlinear. The I–V curve included both areas of inward and outward rectification. The dependence of inward rectification upon the K⁺ ion electrochemical gradient was demonstrated. The membrane was capable of a regenerative action potential due to permeability changes for Ca²⁺ and Na⁺ ions. The Ca and Na components of the action potential were identified. The Ca component was reversibly suppressed by cobalt and irreversibly blocked by D-600. The Na component was tetrodotoxin (TTX) insensitive. The excitable response of P. miniata oocytes is similar to that described by Miyazaki et al. (1975a) for those of the starfish Asterina pectinifera.Immature oocytes were stimulated to mature with 10^?5M 1-methyladenine (1-MA) during continuous monitoring of the membrane potential. The resting potential in ASW became more inside negative during maturation. This change of the passive membrane property of the oocyte may be accounted for by the increased selectivity to K⁺ ions. The specific membrane resistance near the resting potential increased from 4.2 ± 1.4 to 21 ± 8.7 kΩ·cm² (n = 15) during maturation, while the specific membrane capacitance decreased slightly from 2 ± 0.5 to 1.7 ± 0.6 μF/cm² (n = 5). Maturation had little effect upon the active membrane properties.     

Regulatory mutants of dihydrofolate reductase in Escherichia coli K12

Summary Trimethoprim inhibits dihydrofolate reductase. Mutations conferring trimethoprim-resistance on E. coli K12 result in either an altered reductase with decreased affinity for the drug, or in 2–30 fold higher levels of the enzyme. Studies of the latter class of mutants indicate that dihydrofolate reductase is regulated by a diffusible molecule, and is probably under negative control. The regulatory mutants, some of which are temperature-sensitive, act cis.     

Factors Limiting the Number of Radiation-Induced Chromosome Exchanges : I. Distance: Evidence from Non-Interaction of x-Ray— and Neutron-Induced Breaks

Soaked seeds of Vicia faba were exposed to fractionated doses of x-rays or x-rays and fast neutrons. When the two-hit (exchange) chromosome aberrations were scored at the first mitosis of the root tip, it was observed that with short fractionation times the radiation-induced breaks from the two x-ray doses could rejoin with one another to form exchanges in proportion to the square of the total dose. If, however, one dose was x-rays and the second neutrons, then no quantitatively determinable interaction occurred between the breaks induced by each of the doses, and the aberration yield was simply the sum of that induced by each fraction. The phenomenon of non-interaction as observed by these dose fractionation studies and also by the linear dose response curve for two-break aberrations induced by neutrons has led to calculations of the distance over which two breaks can rejoin. The distance is evidently much smaller than the previously accepted value of 1 µ.     

AN ELECTRON MICROSCOPE STUDY OF THE EPITHELIUM IN THE NORMAL MATURE AND IMMATURE MOUSE CORNEA

1. An electron microscope study at high resolution of the corneal epithelium of the normal mature and immature mouse revealed new information regarding the submicroscopic appearance of these cells. 2. Two thin dense lines separated by a less dense area constituted the structure of the limiting surface membrane of epithelial cells; the thickness of this membrane was about 80 A. 3. Some differences in the appearance of the cytoplasm and mitochondria of cells from the immature mouse cornea and the appearance of the cytoplasm and mitochondria of cells from the adult mouse cornea were observed. 4. The basement membrane appeared as a dense band about 600 A wide separating the basal epithelial cells from the substantia propria. Suggestions of periodicity were seen in some phosphotungstic acid-treated specimens. 5. Round bodies believed to be bacteria were seen on the surface of the outer epithelial cells in the adult mouse cornea but not in the immature, unopened eye.     

A resonance Raman study of substrate and inhibitor binding to protocatechuate-3,4-dioxygenase

Resonance Raman spectra were obtained for complexes of protocatechuate-3,4-dioxygenase with substrate and hydroxybenzoate inhibitors. The data establish metal coordination by these bound species and demonstrate further that tyrosine ligation, present in the resting enzyme, is not altered in the complexes. For the inhibitors, 3-chloro-4-hydroxybenzoate and 3-fluoro-4-hydroxybenzoate, the data are interpreted as indicating iron ligation by the phenolate functionality. For the substrate, 3,4-dihydroxyphenylproprionate, chelation via the o-dihydroxy grouping is proposed. In all three complexes tyrosine ligands present in the resting enzyme are not displaced. The inhibitor scattering intensity was utilized as an internal standard to estimate that two tyrosines are coordinated to the iron at the active site.