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Biomedical subjects

M Lederman

Publications and source records attributed to M Lederman.

At least 37 records · Page 2Linked to original sources

Natural parvovirus infection in laboratory rabbits.

Laboratory rabbits from various commercial and private sources were found to have high serum antibody titers specific for lapine parvovirus (LPV). By both immunofluorescence and hemagglutination inhibition assays, 75% of these sera were positive for LPV. This finding, together with the recovery of LPV from kidneys of neonatal rabbits, suggested that LPV infection is common in commercially available rabbits in the United States. It was concluded that use of infected rabbits could interfere with research in which rabbit cell cultures or in vitro immunologic assays are used.

Animals↗

Analysis of the termini of the DNA of bovine parvovirus: demonstration of sequence inversion at the left terminus and its implication for the replication model.

The distribution of terminal-sequence orientations in the viral DNA of bovine parvovirus (BPV), an autonomous parvovirus, was studied by end labeling and restriction enzyme digestion and also by cloning. The left (3') end of the minus strand of BPV was found in two alternative sequence orientations (designated as flip and flop, which are reverse complements of each other), with a 10-fold excess of flip. This is in contrast to the autonomous rodent parvoviruses which encapsidate minus-strand DNA with only the flip orientation at this end. The right (5') end of the minus strand of BPV contained both sequence orientations with equal frequencies, as in the rodent parvoviruses. Sequence inversions were also detected at both ends of the plus strand, which makes up about 10% of the encapsidated BPV DNA. Each terminus of BPV DNA had a characteristic ratio of flip to flop forms, and this ratio was restored in the progeny DNA resulting from transfection with genomic clones of different defined terminal conformations. Replicative-form DNA showed the same distribution of terminal-sequence orientations as the reannealed plus and minus virion DNAs, suggesting that the distribution of flip and flop forms observed in virion DNA is not due to selective encapsidation, but rather to the specific distribution of replicative forms. The current replication model for autonomous parvoviruses, which was based on the available data for the rodent parvoviruses, cannot account for the observed distribution of BPV DNA. An alternative model is suggested.

Animals↗

Genomic clones of bovine parvovirus: construction and effect of deletions and terminal sequence inversions on infectivity.

Genomic clones of the autonomous parvovirus bovine parvovirus (BPV) were constructed by blunt-end ligation of reannealed virion plus and minus DNA strands into the plasmid pUC8. These clones were stable during propagation in Escherichia coli JM107. All clones tested were found to be infectious by the criteria of plaque titer and progressive cytopathic effect after transfection into bovine fetal lung cells. Sequencing of the recombinant plasmids demonstrated that all of the BPV inserts had left-end (3')-terminal deletions of up to 34 bases. DNA isolated from progeny virions arising from transfected infectious clones was found to be indistinguishable from wild-type DNA by restriction enzyme analysis. Defective genomes could also be detected in the progeny DNA even though the infection was initiated with homogenous, cloned DNA. Full-length genomic clones with 3' flip and 3' flop conformations were constructed and were found to have equal infectivity. Analysis of low-molecular-weight DNA isolated from lysates of cells transfected with these clones demonstrated that rescue and replication of BPV DNA could be detected 3 to 8 days after transfection. Expression of capsid proteins from transfected genomes was demonstrated by hemagglutination, indirect immunofluorescence, and immunoprecipitation of [35S]methionine-labeled cell lysates. Use of appropriate antiserum for immunoprecipitation showed the synthesis of BPV capsid and noncapsid proteins after transfection. Independently, a series of genomic clones with increasingly larger 3'-terminal deletions was prepared from separately subcloned 3'-terminal fragments. Transfection of these clones into bovine fetal lung cells revealed that deletions of up to 34 bases at the 3' end lowered but did not abolish infectivity, while deletions of greater than 52 bases were lethal. End-label analysis showed that the 34-base deletion was repaired to wild-type length in the progeny virus.

Amino Acid Sequence↗

Cephalic vein guide wire technique for implantation of permanent pacemakers.

This report describes a modified cephalic vein guide wire technique for the implantation of permanent pacemakers. The procedure was attempted in 11 patients only when pacing leads could not be passed directly into the exposed cephalic vein. Eight bipolar DDD (two leads), two unipolar DDD, and one bipolar VVI (one lead) pulse generators were implanted successfully, with only one relatively minor complication. The cephalic vein guide wire technique provides easy access to the subclavian vein and may allow implantation of most single- and dual-chamber pacemakers with only a cephalic vein cutdown.

Aged↗

Bovine parvovirus DNA-binding proteins: identification by a combined DNA hybridization and immunodetection assay.

We have investigated the interaction between bovine parvovirus (BPV) capsid and non-capsid proteins and restriction fragments of the BPV genome by a combined DNA hybridization and immunodetection assay. 32P-labelled DNA was bound to nitrocellulose membranes bearing lysates of mock-infected and virus-infected cells whose proteins had been separated by SDS-polyacrylamide gel electrophoresis. The position of bound DNA was determined by autoradiography. The proteins on the membrane were still accessible to specific antibodies, allowing confirmation of the DNA-binding species by an immunodetection reaction. In 0.2 M-NaCl, BPV capsid proteins VP2 (72,000 daltons) and VP3 (62,000 daltons) bound the 0 to 16 map unit EcoRI fragment of BPV DNA which contained label in either the minus or plus strand. At higher salt concentration (0.5 M), only VP2 still bound DNA. Within this fragment, the capsid protein binding was restricted to those nucleotides between map units 0 and 4. No binding to capsid proteins was seen with the fragment spanning the middle of the genome and minor binding to VP3 was seen with the 5' end. Binding to the BPV non-capsid protein NP-1 was observed with the 0 to 16 map unit fragment when label was in the virion strand and to other possibly BPV-coded proteins when label was in the plus strand. The NP-1 binding was localized to map units 4 to 16. We did not detect binding to the BPV homologue(s) of the autonomous parvovirus non-capsid protein NS1, due in part to its low concentration in the cell lysates used. Points of the parvovirus replication cycle at which DNA-binding proteins may serve controlling functions are discussed.

Animals↗

Detection of bovine parvovirus proteins homologous to the nonstructural NS-1 proteins of other autonomous parvoviruses.

Two nonstructural proteins of bovine parvovirus (BPV) with apparent molecular sizes of 75,000 and 83,000 daltons have been detected. The proteins were immunoprecipitated from lung cells infected with various isolates of BPV and from in vitro translations of infected cell mRNA. These proteins were expressed as nuclear phosphoproteins and were synthesized early in infection, before the peak of capsid protein synthesis. Early in infection, the 75-kilodalton-size species could be resolved into two bands of equal intensity, but later in infection, the lower-molecular-size form predominated. Antibodies directed against bacterial fusion proteins encoding amino acid sequences from a highly conserved region of the NS-1 polypeptides of two other parvoviruses, minute virus of mice and the human virus B19, gave specific nuclear fluorescence with BPV-infected cells, although the antibodies failed to immunoprecipitate any viral proteins. The noncapsid proteins appear to be homologous to the previously characterized NS-1 proteins of other autonomous parvoviruses.

Animals↗

Possible sequences for nuclear accumulation of parvoviral proteins.

Parvoviral genomes have been searched for sequences which may code for the nuclear transport of viral proteins. Sequences similar to those which regulate the nuclear transport of T antigen and yeast mating type protein were detected within the sequences coding for capsid and non-capsid proteins.

Amino Acid Sequence↗

Complete nucleotide sequence and genome organization of bovine parvovirus.

We determined the complete nucleotide sequence of bovine parvovirus (BPV), an autonomous parvovirus. The sequence is 5,491 nucleotides long. The terminal regions contain nonidentical imperfect palindromic sequences of 150 and 121 nucleotides. In the plus strand, there are three large open reading frames (left ORF, mid ORF, and right ORF) with coding capacities of 729, 255, and 685 amino acids, respectively. As with all parvoviruses studied to date, the left ORF of BPV codes for the nonstructural protein NS-1 and the right ORF codes for the major parts of the three capsid proteins. The mid ORF probably encodes the major part of the nonstructural protein NP-1. There are promoterlike sequences at map units 4.5, 12.8, and 38.7 and polyadenylation signals at map units 61.6, 64.6, and 98.5. BPV has little DNA homology with the defective parvovirus AAV, with the human autonomous parvovirus B19, or with the other autonomous parvoviruses sequenced (canine parvovirus, feline panleukopenia virus, H-1, and minute virus of mice). Even though the overall DNA homology of BPV with other parvoviruses is low, several small regions of high homology are observed when the amino acid sequences encoded by the left and right ORFs are compared. From these comparisons, it can be shown that the evolutionary relationship among the parvoviruses is B19 in equilibrium with AAV in equilibrium with BPV in equilibrium with MVM. The highly conserved amino acid sequences observed among all parvoviruses may be useful in the identification and detection of parvoviruses and in the design of a general parvovirus vaccine.

Amino Acid Sequence↗

Spiroperidol binding sites on mouse lymphoid cells. Effects of ascorbic acid and psychotropic drugs.

Since highly differentiated cells of mammalian immune systems reportedly have binding sites for a variety of neurohumoral agents, we investigated parameters related to possible existence of dopamine receptors on murine lymphoid cells. Using a dopamine antagonist, [3H]spiroperidol, we found evidence for displaceable binding on mouse spleen cells. Total and displaceable (10 microM haloperidol) binding of spiroperidol was markedly enhanced by the absence of ascorbic acid in the incubation medium. Displaceable binding of a dopamine receptor agonist [( 3H]ADTN) could not be found on lymphoid cells either in the presence or absence of ascorbic acid. In the absence of ascorbate, displaceable spiroperidol binding to mouse spleen cells revealed partially saturable, but complex, kinetics and we calculated positive cooperativity at low ligand concentrations.

Animals↗

Virally coded noncapsid protein associated with bovine parvovirus infection.

A phosphorylated protein (NP-1) with an Mr of 28,000 has been detected in nuclei of bovine parvovirus (BPV)-infected cells in association with chromatin. No protein in this size range was detected after infection of appropriate cells with several autonomous rodent parvoviruses although the BPV-specific protein is similar in size to noncapsid proteins associated with rabbit parvovirus or adeno-associated virus infection. Structural homology between NP-1 and a BPV capsid protein could be detected by electrophoretic analysis of the products of proteolysis with chymotrypsin. This protein can be detected after in vitro translation of RNA from BPV-infected cells and BPV-specific RNA. Homology between the in vivo- and in vitro-synthesized species was shown by the similarity of the chymotryptic products.

Animals↗

Malignant epibulbar melanoma: natural history and treatment by radiotherapy.

One hundred and eighty-four cases of malignant melanoma of the lids and epibulbar region were seen in the Radiotherapy Department of the Royal Marsden Hospital during the period 1943-74. Some of these tumours were radiosensitive and could thus be treated by radiotherapy without loss of the affected eye. Their clinical and histological features are described. The treatment policy adopted was based on radiotherapy first, and surgery reserved for the failures. A policy of observation is advised for precancerous melanosis, and active treatment should be undertaken only if malignancy supervenes. The contraindications for radiotherapy, the factors governing radiosensitivity, and the incidence and site of metastases are reported, and the possible effect on the metastatic and survival rates of leaving the eye are discussed. Active treatment of the node-free neck is not advised. Results are given according to site, histology, and treatment method, and the frequency with which the eye was lost is shown, since the sole advantage of radiotherapy over surgery is the possibility of saving the eye without worsening the prognosis for survival.

Adult↗

In vitro and in vivo studies of bovine parvovirus proteins.

Total cytoplasmic RNA from bovine parvovirus (BPV)-infected cells or BPV-specific RNA selected by hybridization to cloned BPV genomic sequences were translated in a message-dependent rabbit reticulocyte lysate. Immunoprecipitation, using immunoglobulin G from rabbits injected with purified BPV, resulted in the detection of [35S]methionine-labeled polypeptides with MrS of 80,000, 72,000, 62,000, and 60,000. These in vitro translation products had the same mobility on sodium dodecyl sulfate-polyacrylamide gels as that of the four proteins found in purified virions. The three largest polypeptides had amino acid sequence homology, as judged by serological methods and partial proteolysis with Staphylococcus aureus V8 protease. Additional noncapsid proteins with MrS of 25,000, 27,000, and 31,000 were also detected as translation products of these RNAs. All of the above species were immunoprecipitated by immunoglobulin G from a calf which was naturally infected with BPV. All four capsid proteins but only one of the lower-molecular-weight polypeptides were detected after the immunoprecipitation of BPV-infected cells. The results presented here indicate that the BPV genome codes for four capsid proteins and a noncapsid protein which may be structurally related to the capsid proteins.

Animals↗

Elastin fibers in the sputum of patients with necrotizing pneumonia.

We have observed five patients for whom the presence of fibers of elastin in potassium hydroxide (KOH) preparations of sputum represented the first evidence of necrotizing pulmonary disease. In four cases, the discovery of elastin fibers in sputum provided additional evidence supporting initiation or modification of antibiotic therapy. Necrotizing disease was confirmed in all cases by autopsy or by the development of cavitation on chest x-ray film. Cytochemical staining, electron microscopy, and elastase digestion all suggest that the refractile fibers seen on KOH wet mount of sputum are elastin. The test, first described in 1846, is simple to perform, requires little experience to read, and may be a valuable adjunct to the chest roentgenogram in the diagnosis of pulmonary parenchymal destruction.

Adult↗

Cancer of the oropharynx: classification and natural history.

The development, comparative anatomy, and function of the oropharynx are briefly described. The text book descriptions of the anatomical boundaries of the oropharynx are commented on and an attempt is made to define these boundaries more rigidly and accurately. Although the advantages and limitations of the UICC classification of oropharyngeal cancer is accepted, an alternative classification is suggested based on the function of the site involved rather than its descriptive anatomy. This has resulted in a necessary rearrangement of the tumor sites and subsites. Subdivision into three sites is suggested: 1. An anterior or glosso-epiglottic; 2. Lateral or tonsillar; 3. Postero-superior or pharyngoepiglotti. The main change has been in the lateral group, the faucial pillars being separated from the tonsil and reallocated. Three different forms of tonsillar cancer are described in some detail; the linguo (glosso) tonsillar sulcus receives mention. A general account is given of the remaining anterior and posterior sites.

Epiglottis↗