EFFECT OF CYANIDE AND ELECTRICAL STIMULATION ON PHOSPHOINOSITIDE METABOLISM IN LOBSTER NERVES

The contents of phosphoinositides, ATP, glucose and lactate in leg and claw nerves of the lobster were determined. Nerves were also analysed after cyanide poisoning, after electrical stimulation, and 1 h after removing the leg from the lobster. Cyanide poisoning decreased the levels of ATP and glucose and increased the content of lactate but did not alter the levels of phosphoinositides. Nerves left in situ for 1 h after disconnection from the central nervous system exhibited a decrease in the content of tri-phosphoinositides (TPI) of 50 per cent, without changes in ATP, glucose or lactate. The TPI change was reversed after incubation for 1 h in oxygenated seawater. Nerves labelled in vivo with ³²P were removed and stimulated at 50 Hz for 5 min. The turnover of TPI phosphorus increased on stimulation in both normal and cyanide-poisoned nerves. In contrast, turnover of ATP increased after stimulation in normal nerves but not in cyanide-treated nerves. We sought to determine whether polyphosphoinositides play a greater role in resting metabolism of the nerve or in the conducting mechanisms. Our results make more likely the involvement of TPI in permeability changes of neural membranes during excitation.     

Properties of Hamster Embryo Fibroblasts Transformed In Vitro After Exposure to Ultraviolet-Irradiated Herpes Simplex Virus Type 2

An in vitro method which led to the transformation of hamster embryo fibroblasts after exposure to herpes simplex virus type 2 (HSV-2) inactivated with ultraviolet irradiation is described. The transformed cells (333-8-9) produced tumors when inoculated into newborn Syrian hamsters but not when injected into weanling Syrian hamsters of the same LSH inbred strain. However, after one in vivo passage, the 333-8-9 cells became highly oncogenic in weanling hamsters. No infectious virus was recovered from these cells. Herpes simplex virus antigens were detected in the transformed cells by the indirect immunofluorescence technique. Sera from tumor-bearing hamsters contained antibody with highly specific neutralizing activity against HSV-2. These studies indicate the continued involvement of the HSV-2 genome in an oncogenic cell line.     

NUCLEAR-CYTOPLASMIC RELATIONS IN THE MITOSIS OF SEA URCHIN EGGS : III. γ-Ray-Induced Damage to Whole Eggs and Nucleate and Anucleate Half-Eggs

Sea urchin eggs were cut into halves. The nucleate and anucleate halves and whole eggs were irradiated with γ-rays and then fertilized with normal sperm. The first mitosis of the diploid half-egg was more delayed than the division of the whole egg. There was a small, but highly significant, delay of the mitosis of the haploid half-egg, thus demonstrating cytoplasmic sensitivity to ionizing radiation. Since the sensitivity of nucleate cells is influenced by cytoplasmic volume, the problem of the role of cytoplasm in repair is considered in relation to these data and other reports in the literature.     

THE BINDING OF BOTULINUM TOXIN TO MEMBRANE LIPIDS: SPHINGOLIPIDS, STEROIDS AND FATTY ACIDS

A number of lipids known to be constituents of nerve-ending membranes were tested for their ability to inactivate botulinum toxin. Inactivation of the toxin by a lipid was taken as presumptive evidence that the lipid might be the in vivo receptor for the toxin. Several sphingolipids (sphingosine, galactosylceramide, glucosylceramide, lactosylceramide, cytolipin K and cytolipin R), steroids (cholesterol and deoxycholic acid) and fatty acids (palmitic acid, stearic acid, prostaglandin E₁) did not affect the potency of botulinum toxin, and thus were discounted as potential toxin receptors. However, the gangliosides did inactivate botulinum toxin rapidly (in less than 5 min), within a temperature range of 2°-40°C, and at ionic strengths of 0.05-0.40. Inactivation diminished as pH fell below 6. The activity of gangliosides in suppressing the potency of botulinum toxin was a function of the number of sialic acid residues in the lipid. Thus, the data suggest that a molecule containing sialic acid may be the receptor for the toxin.     

Applicability of radiotracer methods of measuring14CO2 assimilation for determining microbial activity in soil including a newin situ method

The applicability of two methods (pyrolysis gas chromatography and acidification-wet oxidation) for determining¹⁴CO₂ incorporation into soil microorganisms was investigated. Both methods were able to distinguish biologically incorporated¹⁴C from abiotically adsorbed¹⁴C, but to varying degrees, there being a larger carryover of abiotic¹⁴C into the organic fraction and a higher percentage of assimilated¹⁴C in the organic fraction with the wet oxidation method. Using¹⁴C assimilation as a measure, it was possible to determine microbial activities in soils of diverse properties under a variety of conditions, including polar soils under harsh environmental conditions. Both light and dark¹⁴CO₂ fixation was measurable.¹⁴CO₂ assimilation was not always proportional to the enumerable microorganisms. A new design for measurement of microbial activityin situ enabled measurement of total C influx (primary productivity) into soils with minimal perturbation to the natural soil ecosystem.     

Lymphangiography: Its Practical Value in Surgically Staged Patients with Hodgkin's Disease

Since the introduction of “staging laparotomy” (to determine the disease''s stage) in assessing Hodgkin''s disease, some observers have argued that lymphangiography could be safely omitted in the initial diagnostic evaluation.To test these opinions a series of 75 patients with Hodgkin''s disease who had a staging laparotomy and histological correlation with lymphangiograms was reviewed. Of 16 examinations with positive results, one proved to be a false positive. Of the 14 examinations with equivocal results, one proved histologically positive. In the remaining 45 lymphangiograms, five were falsely negative. In all five of these patients abdominal lymph nodes were involved, but in areas that do not routinely opacify on lower extremity lymphangiography. The overall accuracy was 90 percent.Therapeutically, the lymphangiogram permits accurate planning for treatment by radiation therapy so that all known disease is treated and yet bone marrow is not excessively irradiated. Changes in lymph node architecture after therapy provide valuable information as to regression of the disease or signs of its early recurrence.     

Preliminary characterization of Thy-1.1 and Ag-B antigens from rat tissues solubilized in detergents

1. A radioactive binding assay for Thy-1.1 alloantigen which functions in the presence of detergents was established by using glutaraldehyde-fixed thymocytes as target cells. Thy-1.1 activity in detergent extracts was then assayed by measuring inhibition of the binding assay. 2. Solubilization of Thy-1.1 from whole thymocytes, and their membranes by a large number of non-ionic detergents and deoxycholate was studied. In the same extracts Ag-B(4) histocompatibility antigenic activities were measured. With the exception of Nonidet P-40, the detergents did not affect the antigenicity of Thy-1.1, but only Lubrol-PX and deoxycholate gave effective solubilization as measured by activity remaining in the supernatant after centrifugation at 200000g for 40min. With Ag-B(4) antigen, Triton X-100, Triton X-67 and Nonidet P-40 gave effective solubilization as well as Lubrol-PX and deoxycholate. Solubilization of Thy-1.1 activity from leukaemia cells and a brain homogenate was also studied, but none of the non-ionic detergents gave satisfactory results with these tissues. 3. Extracts from thymocyte membranes were further examined by gel filtration and sucrose gradient centrifugation. The Thy-1.1 activity behaved as a single component in deoxycholate with a density similar to that of a globular protein, but in Lubrol-PX the antigen was contained in a low-density complex. In Lubrol-PX extracts Ag-B(4) was also found in aggregates not observed in deoxycholate. 4. The s(20,w) values for Thy-1.1 and Ag-B(4) antigens in deoxycholate were 2.4 and 4.4, and v values were 0.70 and 0.75 respectively. The Stokes radius observed for Thy-1.1 was 3.1nm and for Ag-B(4) 5.3nm. By using these values the molecular weights for the antigen-detergent complexes were calculated to be 28000 for Thy-1.1 and 100000 for Ag-B(4).