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C Hurwitz

Publications and source records attributed to C Hurwitz.

At least 37 records · Page 2Linked to original sources

Localization of polyamine enhancement of protein synthesis to subcellular components of Escherichia coli and Pseudomonas sp. strain Kim.

At 5 mM Mg2+, spermidine stimulation of polyphenylalanine synthesis by cell-free extracts of Escherichia coli was found to be about 30 times greater than that by extracts of Pseudomonas sp. strain Kim, a unique organism which lacks detectable levels of spermidine. By means of reconstitution experiments, the target of spermidine stimulation was localized to the protein fraction of the highspeed supernatant component (S-100) of E. coli and was absent from, or deficient in, the S-100 fraction of Pseudomonas sp. strain Kim. The spermidine stimulation did not appear to be due to the presence in the E. coli S-100 fraction of ribosomal protein S1, elongation factors, or E. coli aminoacyl-tRNA synthetases. The failure to observe spermidine stimulation by the Pseudomonas sp. strain Kim S-100 fraction was also not due to a spermidine-enhanced polyuridylic acid degradation. The synthesis of polyphenylalanine by Pseudomonas sp. strain Kim extracts was stimulated by putrescine and by S-(+)-2-hydroxyputrescine to a greater degree than was synthesis by E. coli extracts. The enhancement by putrescine and by S-(+)-2-hydroxyputrescine with Pseudomonas sp. strain Kim extracts was found to be due to effects on its ribosomes.

Bacterial Proteins↗

T-lymphocyte subsets in smoking and lung cancer: Analysis of monoclonal antibodies and flow cytometry.

In order to determine whether abnormalities of immunoregulatory T-cells occur in patients with lung cancer, we characterized peripheral T-lymphocytes in 26 patients with untreated lung cancer. The results in patients with primary squamous cancer (SC) (n = 10), primary adenocarcinoma (AC) (n = 7), and secondary lung metastases (M) (n = 9) were compared with each other and to subjects without cancer (n = 48), including nonsmokers (n = 29) and smokers (n = 19). We found that OKT3+ (mature, peripheral (T-lymphocytes, including both OkT4+ (inducer/helper) and OKT8+ (cytotoxic/suppressor) lymphocytes, were increased in light-to-moderate smokers, but that OKT4+ cells were decreased n heavy smokers (p less than 0.05). The ratio of OKT4+ to OKT8+ (4/8) lymphocytes, reflecting the balance of immunoregulatory cells, was normal in light-to-moderate smokers, but was decreased in heavy smokers (p less than 0.05). The profile of circulating T-cells in patients with SC was similar to the smokers. In contrast, in patients with AC, we found a decreased percentage of OKT8+ cells (p less than 0.05). The 4/8 ratio was elevated in patients with AC (p less than 0.05). In patients with M, there was a decreased percentage of OKT3+ cells reflected in both OKT4+ and OKT8+ subsets. The 4/8 ratio in patients with M was low. Thus, a number of abnormalities in circulating T-cells was found both in smokers and in patients with lung cancer. These results suggest that immunoregulatory abnormalities contribute to the pathogenesis of lung cancer.

Adenocarcinoma↗

Role of ribosome recycling in uptake of dihydrostreptomycin by sensitive and resistant Escherichia coli.

Exposure of streptomycin-resistant cells to puromycin results in uptake of dihydrostreptomycin comparable to that found with streptomycin-sensitive cells. This finding indicates that the enhanced phase of uptake, previously reported only in sensitive cells, may result from an increase in internal binding sites, presumably run-off ribosomes. The increased uptake of dihydrostreptomycin resulting from exposure to puromycin is greatest in both sensitive and resistant cells at concentrations below 100 microgram/ml. At 100 microgram/ml, exposure to puromycin in vivo results in significant, but not complete, polysome degradation and inhibition of protein synthesis. At 500 microgram/ml, where polysome degradation is complete in less than 2 min and where growth and protein synthesis are inhibited more than 90%, uptake of dihydrostreptomycin by both sensitive and resistant cells is inhibited. Puromycin has no effect on binding of dihydrostreptomycin to 70-S monosomes, as measured by equilibrium dialysis. The increased uptake of dihydrostreptomycin by resistant cells resulting from exposure to puromycin has no effect on viability. Addition of N-ethylmaleimide immediately and completely inhibits the puromycin-induced uptake of dihydrostreptomycin even when added after substantial polysome degradation has occurred.

Dihydrostreptomycin Sulfate↗

Determination of C1q in human sera.

All detectable C1q in serum is precipitated with the euglobulin fraction, and all other detectable hydroxyproline-containing protein in serum is excluded from this fraction. Since C1q contains 4.3% hydroxyproline, an estimate of its content in serum can be calculated by multiplying the hydroxyproline content of the euglobulin fraction by 23.3. The mean value of C1q in five normal male subjects determined by this method was found to be 63 microgram/ml as compared with 144 microgram/ml when determined by RID, indicating that the latter procedure seriously overestimates C1q in serum. C1q content of serum from patients with RA was found to be elevated by both procedures.

Arthritis, Rheumatoid↗

Newly discovered polyamine, 2-hydroxyspermidine, in Pseudomonas acidovorans.

A previously unknown hydroxylated polyamine has been recovered from Pseudomonas acidovorans 29. It has been identified as 2-hydroxyspermidine, N4-(3-aminopropyl)-1,4-diaminobutane-2-ol, by its chromatographic behavior, electrophoretic mobility, and reaction with metaperiodate. It can be synthesized enzymatically from 2-hydroxyputrescine by cell-free preparations from Escherichia coli or P. acidovorans 29 which contain propylamine transferase. It is interesting to note that the naturally occurring compound is the 2-hydroxyspermidine and not the 3-hydroxyspermidine, N1-(3-aminopropyl)-1,4-diaminobutane-2-ol, indicating that the propylamine transferase reacts preferentially with the amine distal to the hydroxyl group. A mixture of 2- and 3-hydroxyspermidines and hydroxyspermine was synthesized by reacting acrylonitrile with 2-hydroxyspermidine and catalytic reduction of the products with hydrogen. N-(gamma-aminopropyl)-beta-alanine, used to help identify the hydroxyspermidines, was synthesized from N-(3-aminopropyl)-3-aminopropanenitrile by hydrolysis with 10% NaOH.

Pseudomonas↗

Antagonistic action between spermidine and putrescine on association and dissociation of purified, run-off ribosomes from Escherichia coli.

The effects of polyamines on the equilibrium between prokaryotic ribosomal subunits and 70 S ribosomes have been studied as a function of concentration of Mg2+ from 2.5 to 7.5 mM. Run-off ribosomes were obtained from Escherichia coli and were washed with buffered 1 M NH4C1. Spermidine at 1 mm favors association of subunits at all concentrations of Mg2+. Putrescine, at concentrations above 8 mM, favors net dissociation at concentrations of Mg2+ below 4.5 mM. Streptomycin behaves like spermidine, while putrescine behaves like initiation factor 1 and initiation factor 3. The effect of putrescine on dissociation is time-dependent and appears to have a half-life of about 3.5 min at 30 degrees. When added after the effects of spermidine or streptomycin on association have occurred, putrescine still causes dissociation. The data suggests that putrescine may reduce net formation of vacant 70 S ribosomes. Another possibility is that putrescine and spermidine may act antagonistically to maintain a labile equilibrium between ribosomal subunits and vacant 70 S ribosomes. It may be significant that the putrescine effect is observed at the concentration of Mg2+ found to be optimum for initiation.

Escherichia coli↗

Separation of hydroxyproline-containing protein from C1q (a subcomponent of complement) in serum.

Precipitation of the euglobulin fraction from serum separates C1q from another hydroxyproline-containing protein(s), which remains in the serum-minus-euglobulin fraction. The presence of C1q in the euglobulin fraction has been verified by radial immunodiffusion assay, by sensitivity to collagenase, by hydroxyproline content, and by electrofocusing. The presence of another hydroxyproline-containing protein(s) in the serum-minus-euglobulin fraction has been verified by the same criteria. The isoelectric pH range of C1q is 6.1-7.0; the isoelectric pH range of the other hydroxyproline-containing protein(s) is 4.2-5.5.

Blood Protein Electrophoresis↗

Effect of tetracycline on puromycin-induced polysome degradation: influence of magnesium and polyamines.

Puromycin-induced polysome degradation has been shown to require G factor, guanosine 5'-triphosphate, and the presence of a ribosome release factor (Hirashima and Kaji, 1972, 1973). Tetracycline, which does not inhibit formation of peptidyl-puromycin (Gottesman, 1967; Sarkar and Thach, 1968) nor the guanosine 5'-triphosphate hydrolysis mediated by elongation factor Tu (Ono et al., 1969), inhibits polysome degradation. The tetracycline inhibition requires Mg(2+) at concentrations above 8 mM, which are inhibitory to protein synthesis in vitro. At concentrations of Mg(2+) below 8 mM, polysome degradation is insensitive to tetracycline, but not to fusidic acid. Addition of spermidine, but not of other polyamines, enables the tetracycline inhibition to occur at concentrations of Mg(2+) as low as 2 mM. The inhibition by tetracycline and by fusidic acid suggests that ribosome movement may be essential for the function of ribosome release factor, or that these antibiotics may directly affect its action.

Escherichia coli↗

Evidence for a magnesium pump in Bacillus cereus T.

Unlike Escherichia coli, Bacillus cereus T appears to accumulate Mg(2+) in its cell sap against a concentration gradient. Over a range of Mg(2+) in the growth medium from 5 x 10(-5) to 1.35 x 10(-2)m, the concentration of Mg(2+) in the cell sap of B. cereus T was maintained at about 6 x 10(-3)m, and ribosome-bound Mg(2+) and spermidine, as well as the spermidine concentration in the cell sap, appear to be unaffected by the concentration of Mg(2+) in the growth medium. Inhibition of growth of E. coli by streptomycin is progressively reversed by increasing the concentration of Mg(2+) in the growth medium above 5 mm. The finding that similar increases of Mg(2+) in the growth medium did not reverse the inhibition of B. cereus T is also consistent with the conclusion that B. cereus T, unlike E. coli, accumulates Mg(2+) to a constant concentration in its cell sap.

Amines↗

Loss of colicinogeny in Escherichia coli strains infected by certain resistance factors.

The original observation that in wild-type colicinogenic Escherichia coli strains the introduction of some R factors abolish their colicin production was studied in certain col(+) strains bearing well-defined col factors. Two resistance (R) factors were used and introduced by conjugation in these strains, namely the 222 factor of Watanabe and a Salmonella typhimurium ST factor (coding for resistance to streptomycin and tetracycline only). The introduction of above mentioned R factors abolished the colicin production of col(+) strains most probably by elimination of col factors. All col factors, however, were not equally susceptible to elimination by the R factors tested, since colicin production in ML strains was abolished by infection by the 222 factor but not by the R factor of S. typhimurium ST, which is able to eliminate other col factors.

Chloramphenicol↗