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S Salzberg

Publications and source records attributed to S Salzberg.

At least 19 recordsLinked to original sources
PubMed1992

Predicting protein secondary structure with a nearest-neighbor algorithm.

We have developed a new method for protein secondary structure prediction that achieves accuracies as high as 71.0%, the highest value yet reported. The main component of our method is a nearest-neighbor algorithm that uses a more sophisticated treatment of the feature space than standard nearest-neighbor methods. It calculates distance tables that allow it to produce real-valued distances between amino acid residues, and attaches weights to the instances to further modify the the structure of feature space. The algorithm, which is closely related to the memory-based reasoning method of Zhang et al., is simple and easy to train, and has also been applied with excellent results to the problem of identifying DNA promoter sequences.

Algorithms↗
PubMed1992

Persistent measles virus infection enhances major histocompatibility complex class I expression and immunogenicity of murine neuroblastoma cells.

The effect of persistent measles virus infection on the expression of major histocompatibility complex (MHC) class I antigens was studied. Mouse neuroblastoma cells C1300, clone NS20Y, were persistently infected with the Edmonston strain of measles virus. The persistently infected cell line, NS20Y/MS, expressed augmented levels of both H-2Kk and H-2Dd MHC class I glycoproteins. Activation of two interferon(IFN)-induced enzymes, known to be part of the IFN system: (2'-5')oligoadenylate synthetase and double-stranded-RNA-activated protein kinase, was detected. Measles-virus-infected cells elicited cytotoxic T lymphocytes that recognized and lysed virus-infected and uninfected neuroblastoma cells in an H-2-restricted fashion. Furthermore, immunization of mice with persistently infected cells conferred resistance to tumor growth after challenge with the highly malignant NS20Y cells. The rationale for using measles virus for immunotherapy is that most patients develop lifelong immunity after recovery or vaccination from this infection. Patients developing cancer are likely to have memory cells. A secondary response induced by measles-virus-infected cells may therefore induce an efficient immune response against non-infected tumour cells.

2',5'-Oligoadenylate Synthetase↗
PubMed1992

Biological effects of photoactivated-HPD and cholesteryl hemisuccinate on erythroid differentiation.

In order to establish the ability of porphyrins to distinguish between differentiated and undifferentiated cells of the erythropoietic pathway, the cytotoxic effects of hematoporphyrin derivative (HPD) on Friend erythroleukemic cells (FLC) were derivative (HPD) on Friend erythroleukemic cells (FLC) were studied. Since cholesterol affects the fluidity of cell membranes, the inhibitory effect of cholesteryl hemisuccinate (CHS) on HPD was similarly tested. FLC were induced with either dimethyl sulfoxide (DMSO), hemin, or both. The cells responded to DMSO-treatment by the synthesis of a large amount of hemoglobin and a decrease in both the cell volume and rate of cell-growth. Hemin, on the other hand, did not drive FLC to synthesize hemoglobin and reduced only moderately the growth rate compared to untreated cultures. The combined effect of DMSO and hemin led to a profound inhibition of the growth rate, but did not increase the synthesis of hemoglobin compared to the level observed in cells treated with DMSO only. The binding property of HPD to FLC was also determined. DMSO-treatment significantly reduced the amount of HPD-binding, and combined treatment with DMSO and hemin resulted in even a lower level of HPD-binding. On the other hand, induced as well as non-induced cells showed the same sensitivity to photoactivated HPD when both DNA and protein synthesis were examined. Pretreatment of both differentiated and undifferentiated cells with CHS reduced the cytotoxicity of photoactivated HPD to the same level. We conclude that the decreased binding of HPD to differentiated cells is a result of a reduction in their cell volume. Thus, the differentiation process per se does not protect cells against the photodynamic effect of HPD. Moreover, CHS may act as a scavenger or first acceptor of singlet oxygen blocking the photodynamic effect.

Animals↗
PubMed1990

Activation of the interferon system during myogenesis in vitro.

Differentiation of skeletal muscle involves withdrawal of myoblasts from cell replication, fusion to form multinucleated myotubes, coordinate appearance of a variety of muscle-specific proteins and the disappearance of a set of other proteins responsible for cell growth. The possible activation of the interferon (IFN) system in this process was studied. Thus, the activity of two IFN-induced enzymes known to be part of the system-(2'-5') oligoadenylate synthetase (2-5A synthetase) and double-stranded RNA-activated protein kinase as well as the expression of 2-5A synthetase coding genes were examined during myogenesis. It is demonstrated that the activity of the enzymes is transiently increased in cultured myoblasts, reaching a peak activity on the 3rd day in culture and then declining to a basal level. This peak activity precedes both cell fusion and the appearance of muscle-specific proteins--acetylcholine receptors (AChR) and creatine kinase. The same kinetics of 2-5A synthetase activity was evident in myoblasts from chick, rat or mouse origin. The enzymatic product appears to be primarily the trimer form of 2-5A, rather than a set of oligomers observed in enzymatic reactions performed on IFN-treated cells, including muscle cultures. The kinetics of 2-5A synthetase gene expression revealed that the largest amount of specific RNA transcripts appeared on the 1st day after seeding, followed by a reduction thereafter. In addition, a decrease was also observed in expression of c-myc, a cell-growth-associated protooncogene. However, an increase towards the 2nd day of both AChR and myosin light chain gene expression was evident, indicating selective regulation of gene expression during myogenesis.

2',5'-Oligoadenylate Synthetase↗
PubMed1990

The activity of 2,5-oligoadenylate synthetase, an interferon-induced enzyme, is coupled to the differentiation state of mouse condylar cartilage.

The enzyme 2,5-oligoadenylate synthetase (2-5A synthetase) is associated with the interferon system, with special reference to the differentiation process of various cell types. The present study investigated whether 2-5A synthetase is also involved in the differentiation of neocartilage in perinatal mice. The cartilage of the mandibular condyle, a secondary type of cartilage, develops relatively late in prenatal life; and consequently it was possible to obtain a relatively embryonic cartilage at a developmental stage that could be manipulated enzymatically, in order to separate and thereby obtain its undifferentiated, proliferative portion along with its more mature fraction. Immunohistochemical studies using antibodies against type I and type II collagen and cartilage-specific proteoglycans could have determined the differentiation status of various portions of the developing condyle. However, the above methodology lacks the necessary precision and accuracy to indicate subtle changes in cellular differentiation. It became evident that the activity of 2-5A synthetase was indeed different in cellular compartments that were at different stages of differentiation. In the neonatal condyle the highest level of activity was encountered in proliferating and as yet undifferentiated prechondrocytes, whereas fully differentiated chondrocytes showed a marked decrease in the activity of this enzyme.

2',5'-Oligoadenylate Synthetase↗
PubMed1990

Involvement of interferon-system in the regulation of cell growth and differentiation.

In this report we review the current knowledge on the involvement of the interferon (IFN) system in the regulation of cell growth and differentiation. We also summarize our own data which provide evidence for the strong correlation between IFN-mediated growth-arrest of transformed cells and the elevated enzymatic activity of an IFN-induced protein. Similarly, it is demonstrated that elevated levels of IFN-induced proteins accompany the early phases of in-vitro cell differentiation. IFN-treatment of NIH/3T3 mouse fibroblasts transformed by Moloney-murine sarcoma virus (MSV) resulted in a significant reduction in the rates of cell growth, protein synthesis and cloning efficiency. In parallel, 2-5A-synthetase activity was induced ten-fold above the background level. Treatment of these cells for 3 days with 450 international units (IU)/ml of IFN followed by its removal, resulted in a gradual increase in all parameters associated with cell growth while the 2-5A-synthetase activity was reduced to its normal level. However, almost no recovery occurred when cells were treated with 1,800 IU/ml. In parallel, 2-5A-synthetase activity remained highly elevated even at 3 days after the removal of IFN. In these cells, the expression of both c-myc and v-mos was reduced rapidly following IFN treatment. Upon removal of IFN after 24 h of treatment, the expression of both genes was resumed but with a different kinetics, suggesting that different mechanisms are responsible for the reduction in gene expression. In rat skeletal muscle cultures which differentiate to form myotubes, the level of both 2-5A-synthetase and protein kinase activities was transiently elevated, reaching a peak at 3 days followed by a decrease to background levels. This peak activity precedes the appearance of the major muscle differentiating proteins.

2',5'-Oligoadenylate Synthetase↗
PubMed1990

Reversibility of the antiproliferative effect of interferon.

The reversibility of the antiproliferative effect of interferon (IFN) and its correlations to the induction of (2',5') oligoadenylate synthetase (2-5A synthetase) activity was studied on NIH/3T3 cells transformed by Moloney murine sarcoma virus. The cells were treated with various doses of mouse beta-IFN. At 72 h after treatment, the cultures were subdivided. While half received fresh doses of IFN, the second half received no IFN. Reversibility of the IFN effect was then followed. Three different parameters as indicators for cell proliferation were used: cell growth, protein synthesis and cloning efficiency. In parallel, the IFN-induced activity of 2-5A synthetase was determined. The data obtained led to the following conclusions. (1) The antiproliferative effect of IFN increases with increased IFN concentration (90-1,800 IU/ml) and with time of treatment, up to 72 h after treatment. (2) The induced activity of 2-5A synthetase increases with a much faster rate, reaching maximum activity at 24 h after treatment with 450 IU/ml. This means that the induction of the enzyme precedes the antiproliferative effects of IFN. (3) There is almost no recovery of the IFN antiproliferative effect following treatment for 72 h with high doses of IFN (1,200-1,800 IU/ml). However, at lower doses, recovery is evident. (4) Removal of IFN after treatment for 3 days with 450 IU/ml resulted in a gradual decrease of 2-5A synthetase activity, reaching the basal level at 72 h after removal. However, there is no reduction of enzyme activity following treatment for 72 h with 1,800 IU/ml of IFN.

2',5'-Oligoadenylate Synthetase↗
PubMed1989

Combined treatment with Pseudomonas aeruginosa toxin and interferon on mouse and human cells.

The combined biological effect of Pseudomonas toxin and beta-interferon on mammalian cells was studied on two cell lines. The first was a virus-producing clone derived from NIH/3T3 mouse fibroblasts transformed by Moloney murine sarcoma virus. The second was a clone derived from human amnion cells. The parameters examined were either retrovirus release from the mouse cells or the rate of protein synthesis in both cell lines. When applied together with Pseudomonas toxin, interferon inhibits virus release even at a Pseudomonas toxin concentration that by itself does not exhibit any biological effect on NIH/3T3 cells. This enhancement phenomenon is both Pseudomonas toxin and interferon dose-dependent. Likewise, the combined treatment of either mouse or human cells with Pseudomonas toxin and the appropriate species-specific interferon, inhibits protein synthesis to a much greater extent than either of these agents alone. The kinetics of the inhibition of virus release is different from that seen with protein synthesis indicating that the enhancement phenomenon observed on virus release is not a result of the inhibition of total cellular protein synthesis. Interferon potentiates the effect of Pseudomonas toxin in a species-specific manner, thus suggesting that this process does not occur at the level of cell receptors but is a consequence of a subsequent intracellular event. It is concluded that the enhancement phenomenon does not reflect a direct interaction between interferon and Pseudomonas toxin, since Pseudomonas incubated together with interferon retained its normal biological activity as indicated by the ability of the toxin molecule to transfer the adenine diphosphoribose (ADP-ribose) moiety of nicotinamide-adenine dinucleotide (NAD) onto elongation factor 2 (EF-2).

ADP Ribose Transferases↗
PubMed1989

2'-5'-oligoadenylate synthetase gene expression in normal and murine sarcoma virus-transformed NIH 3T3 cells.

Mouse fibroblasts transformed by murine sarcoma virus (MSV) are highly sensitive to the antiproliferative effect of interferon (IFN) (M. Bakhanashvili, D. H. Wreschner, and S. Salzberg, Cancer Res. 43:1289-1294, 1983). To elucidate the mechanism leading to this IFN sensitivity, the expression of the 2'-5'-oligoadenylate synthetase (2-5A synthetase) gene, the presence of the 2-5A synthetase protein, and the level of its enzymatic activity were determined in IFN-treated and untreated cultures. NIH 3T3 mouse fibroblasts were compared with two different NIH 3T3 clones transformed by MSV. Cultures were treated with 300 IU of beta IFN (IFN-beta) per ml for 16 to 24 h. While no detectable 2-5A synthetase-derived transcripts were seen in untreated NIH 3T3 cells, two size classes of RNA transcripts, i.e., 1.7 and 4.2 kilobases, were detected in IFN-treated cultures. Surprisingly, a similar amount of transcripts were present in untreated transformed cells. However, following IFN treatment, an eightfold increase in the level of RNA was readily detected in these cells, with no change in the size classes. Similar results were obtained with the 2-5A synthetase protein, for which three size classes of 42, 71, and 102 kilodaltons were demonstrated by immunoblotting, and with the enzymatic activity, for which again, the highest level was seen in IFN-treated MSV-transformed cultures. The basal level of 2-5A synthetase gene expression in the transformed cells has biological significance since these cells were more resistant to mengovirus infection than NIH 3T3 mouse fibroblasts. Medium collected from transformed cultures failed to induce 2-5A synthetase activity in NIH 3T3 cells. Furthermore, antibodies directed against mouse IFN-beta failed to inhibit 2-5A synthetase activity detected in transformed cultures. These results suggest that at least IFN-beta secretion is not involved in the elevated level of 2-5A synthetase gene expression in these cells.

2',5'-Oligoadenylate Synthetase↗
PubMed1989

Effect of alpha interferon on growth of leukemic and normal hematopoietic progenitors. Synergism with H2 histamine receptor antagonists.

We studied whether the histamine H2 receptor antagonist, cimetidine, potentiates antiproliferative effects of recombinant alpha interferon (IFN alpha) on in vitro clonal growth of certain normal and malignant hematopoietic cells. Cimetidine alone, at therapeutic serum concentrations, was not inhibitory to the cells studied. IFN alpha alone inhibited the growth of HL-60 leukemic cells only at concentrations greater than 1000 U/ml, whereas with cimetidine, inhibition was seen at greater than 1 U/ml of IFN alpha. The enhancing effect of cimetidine on IFN alpha inhibition of clonal growth was neutralized by histamine and was not seen with histamine H1 receptor antagonist. In HL-60 cells, cimetidine also increased the enzymatic activity of (2'-5')-oligoadenylate synthetase, induced by IFN alpha. The combination of cimetidine and IFN alpha had a synergistic inhibitory effect on the growth of leukemic granulocyte-macrophage colony-forming units (CFU-GM) from chronic myeloid leukemia patients, normal CFU-GM, and normal erythroid burst-forming unit (BFU-E) progenitors. These data suggest that cimetidine may play a role in overcoming resistance to IFN alpha therapy in leukemia, but may also increase IFN alpha hematopoietic toxicity.

2',5'-Oligoadenylate Synthetase↗
PubMed1988

Photosensitization of differentiating Friend erythroleukemic cells by hematoporphyrin derivative and the cholesterol effect.

The Friend erythroleukemia cell line was used to study the binding and biological properties of the photosensitizer hematoporphyrin derivative (HPD) on a differentiating system. In addition, the effect of cholesterol hemisuccinate (CHS) enrichment of cell membranes on HPD activity was tested on the same cell system. Differentiation of Friend erythroleukemia cells (FLC) was induced with dimethyl sulfoxide (DMSO) and resulted in a decreased cell volume and an increased rate of hemoglobin synthesis as a function of the duration of DMSO treatment. Differentiated cells seem to bind less porphyrin than their undifferentiated counterparts. Thus, cells treated for 6 days with DMSO bound 30-40% less dye than an identical number of untreated FLC. In contrast, a similar inhibition of both DNA and protein synthesis by photoactivated HPD was evident in either DMSO-treated or untreated FLC. Enrichment of cell membranes with CHS led to the same degree of protection from the damaging activity of the photoactivated dye in both differentiated and undifferentiated FLC. The decreased binding of HPD to DMSO-treated FLC is most likely a result of a reduction in cell volume of differentiated cells and is not related to an intrinsic property of the differentiation process.

Animals↗
PubMed1988

The psychiatric uses of cold wet sheet packs.

The cold wet sheet pack is a treatment that is seldom discussed anymore. The authors present results of a national survey which demonstrated that this treatment is rarely used in modern American psychiatry. They retrospectively review its recent use for 46 hospitalized psychiatric patients and conclude that the treatment is safe and has interesting and useful effects that go beyond the concept of simple restraint. Further study of treatment with cold wet sheet packs is recommended before it disappears altogether.

Adolescent↗
PubMed1985

Serum interferon as a biological marker in malignant tumors.

A total of 745 serum interferon (IFN) determinations were carried out on the serum of 332 cancer patients with a variety of tumors. Disease activity was defined as either 'active' or 'nonactive' as based on clinical examination, the results of laboratory tests and radiological criteria. IFN levels were estimated for both disease activity and mode of treatment. A statistically significant higher level of IFN in the serum of active, as compared to that of nonactive disease, was established. Furthermore, a slight increase in the IFN level was observed in patients before initiating chemotherapy as compared to untreated patients, followed by a decrease in those patients undergoing chemotherapy. These differences between groups were not found to be statistically significant. However, in the cases of rectum, prostate and female genital cancers, a significant elevated level of serum IFN was observed at a late stage of the disease, followed by a decrease after chemotherapy treatment. In addition, a significant lower level of serum IFN was detected in breast cancer patients shortly after radiotherapy as compared to those before radiotherapy. We conclude that IFN, known to play a role in the control mechanisms of developing malignant processes, can be determined in the serum and used to monitor disease activity in individual patients. The inhibition of IFN release depends in part on therapy and can be used for monitoring treatment schedules along the course of disease.

Antineoplastic Agents↗
PubMed1985

Protective effect of cholesterol on Friend leukemic cells against photosensitization by hematoporphyrin derivative.

The cytotoxic effects of hematoporphyrin derivative (HPD) on Friend erythroleukemic cells were studied. Upon binding of the porphyrin to the cells, the fluorescence spectra was shifted from 613 to 633 nm regarding the main band and from 676 to 667 nm concerning the secondary band. The kinetics of HPD binding was then determined. Maximum binding already occurred at 60 s after exposure of the cells to HPD. It could be demonstrated that the effect of the photoactivated HPD on cell viability was drug, dose, and light fluorescence dependent. Cellular protein synthesis and Friend virus complex release from the cells were equally inhibited by the photodynamic sensitization of the drug, indicating no specific effect on virus maturation. Since cholesterol affects the fluidity of cell membranes, it was important to study the effect of cholesterol enrichment on the photodynamic sensitization by HPD. It was found that, while a 50% reduction in protein synthesis was monitored following treatment with 20 micrograms of HPD per ml and illumination by a 6-milliwatt white light for 60 s, no inhibition was observed following preenrichment of the cells with 0.5, 1, or 2% of cholesterol hemisuccinate. The same trend of cholesterol protection was demonstrated with longer illumination periods up to 10 min. The protective effect of cholesterol hemisuccinate was also seen using scanning electron microscopy. It is thus concluded that the cholesterol hemisuccinate content of Friend erythroleukemic cell membranes is an important factor in regulating the cytotoxicity of photoactivated HPD.

Animals↗
PubMed1979

Adsorption, penetration, and uncoating of murine leukemia virus studied by using its reverse transcriptase.

A procedure using the virus-associated reverse transcriptase was developed for following the kinetics of adsorption, penetration, and uncoating of murine leukemia virus. Viral adsorption to cell membrane was determined by assaying this enzyme activity in isolated debris of mechanically disrupted cells after infection with murine leukemia virus in the presence of actinomycin D. At 37 degrees C, viral adsorption proceeded at a high initial rate, but after 5 min of incubation with the virus, it gradually slowed down. At 4 degrees C, viral adsorption was slower but proceeded at a linear rate. Intracellular virus was determined by centrifuging the cytoplasmic fraction of the disrupted cells at 105,000 x g for 45 min and assaying reverse-transcriptase activity in the high-speed pellet thus obtained. Sucrose gradient analysis of the enzyme activity recovered from the cytoplasm of infected cells indicated that this activity represented intact virus particles. No appreciable amount of such particles was recovered from the cytoplasm of cells infected at 4 degrees C. This indicates that the virions recovered from the cytoplasm of cells infected at 37 degrees C are indeed intracellular virus particles which penetrated into the cells and not just membrane-bound particles mechanically released to the cytoplasmic fraction during cell disruption. By this procedure intracellular virus was found to accumulate in the cytoplasm, reaching a maximal level within 20 min. The accumulated intracellular virus particles gradually disappeared from the cytoplasm, evidently due to their uncoating which was completed within 80 min.

Adsorption↗
PubMed1979

Viral genome RNA serves as messenger early in the infectious cycle of murine leukemia virus.

When NIH/3T3 mouse fibroblasts were infected with the Moloney strain of murine leukemia virus, part of the viral genome RNA molecules were detected in polyribosomes of the infected cells early in the infectious cycle. The binding appears to be specific, since we could demonstrate the release of viral RNA from polyribosomes with EDTA. Moreover, when infection occurred in the presence of cycloheximide, most viral RNA molecules were detected in the free cytoplasm. Size analysis on polyribosomal viral RNA molecules indicated that two size class molecules, 38S and 23S, are present in polyribosomes at 3 h after infection. Analysis of the polyriboadenylate [poly(rA)] content of viral RNA extracted from infected polyribosomes demonstrated that such molecules bind with greatest abundance at 3 h after infection, as has been detected with total viral RNA. No molecules lacking poly(rA) stretches could be detected in polyribosomes. Furthermore, when a similar analysis was performed on unbound molecules present in the free cytoplasm, identical results were obtained. We conclude that no selection towards poly(rA)-containing viral molecules is evident on binding to polyribosomes. These findings suggest that the incoming viral genome of the Moloney strain of murine leukemia virus may serve as a messenger for the synthesis of one or more virus-specific proteins early after infection of mouse fibroblasts.

Animals↗