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Biomedical subjects

B Lorber

Publications and source records attributed to B Lorber.

At least 55 records · Page 3Linked to original sources

Investigation of an outbreak of listeriosis: new hypotheses for the etiology of epidemic Listeria monocytogenes infections.

From December 1986 to March 1987 an outbreak of Listeria monocytogenes infection occurred in the Philadelphia metropolitan area. A patient-control study showed patients were more likely than controls to have had an ill family member and to have used antidiarrheal medication during the month before their illness. Diet histories showed patients were significantly more likely to have eaten ice cream or salami than were controls, and to have shopped at one grocery store chain. Subtyping of L. monocytogenes isolates of patients showed no predominant strain, and cultures of food products eaten by patients were negative except for Brie cheese eaten by one patient. With no predominant strain of L. monocytogenes in the patients, a common source for this outbreak is unlikely. Thus, the identified risk factors may have been associated with carriage of L. monocytogenes and a coinfecting organism may have precipitated disseminated disease. Possible cofactors should be considered in investigations of future outbreaks of listeriosis.

Adolescent↗

Properties of N-terminal truncated yeast aspartyl-tRNA synthetase and structural characteristics of the cleaved domain.

Cytoplasmic aspartyl-tRNA synthetase from Saccharomyces cerevisiae is a dimer made up of identical subunits of Mr 64,000 as shown by biochemical and crystallographic analyses. Previous studies have emphasized the high sensitivity of the amino-terminal region (residues 1-32) to proteolytic enzymes. This work reports the results of limited tryptic or chymotryptic digestion of the purified enzyme which gives rise to a truncated species that has lost the first 50-64 residues with full retention of both the activity and the dimeric structure. In contrast the larger tryptic fragment is distinguished from the whole enzyme by its weaker retention on heparin-substituted agarose gels. The cleaved N-terminal part presents peculiar structural features, such as a high content in lysine residues arranged in a palindromic fashion. The properties of the trypsin-modified enzyme and of the cleaved amino-terminal region are discussed in relation to the known structural characteristics of aspartyl-tRNA synthetase and of other eukaryotic aminoacyl-tRNA synthetases.

Amino Acyl-tRNA Synthetases↗

A high resolution diffracting crystal form of the complex between yeast tRNAAsp and aspartyl-tRNA synthetase.

Three new crystal forms of the complex between yeast tRNAAsp and aspartyl-tRNA synthetase have been produced. The best crystals, obtained after modifying both purification and crystallization conditions, belong to space group P2(1)2(1)2(1) and diffract to 2.7 A. Unit cell parameters are a = 210.4 A, b = 145.3 A and c = 86.0 A (1 A = 0.1 nm), with one dimeric enzyme and two tRNA molecules in the asymmetric unit.

Amino Acyl-tRNA Synthetases↗

Chronic meningococcal meningitis. An association with C5 deficiency.

A 44-year-old man with a history of presumed meningococcal meningitis 32 years before, presented with a three-month illness, characterized by fever, 13.5-kg (30-lb) weight loss, occipital headache, shoulder pain, and muscle weakness, which had been diagnosed as "polymyositis" and treated accordingly. Evaluation revealed meningococcal meningitis due to serogroup W135 and a C5 deficiency. Evidence for the occurrence of chronic meningococcal meningitis is described.

Adult↗

Changing patterns of infectious diseases.

In the 20 years that have passed since I received my medical degree, some of the most evident, striking, and interesting changes in patterns of diseases have occurred among those due to transmissible agents. Among other changes are the recognition of new diseases, new clinical manifestations for old diseases, new ecologic niches for traditional pathogens, and new modes of disease transmission. Implicated causes for these changes include alterations in lifestyle such as sexual behaviour, leisure activity, and dietary trends, along with the impact of immigration and the effects of medical progress. A review of these changing patterns demonstrates the dynamic nature of medicine and suggests lessons for medical education.

Acquired Immunodeficiency Syndrome↗

The microheterogeneity of the crystallizable yeast cytoplasmic aspartyl-tRNA synthetase.

Yeast aspartyl-tRNA synthetase is a dimeric enzyme (alpha 2, Mr 125,000) which can be crystallized either alone or complexed with tRNAAsp. When analyzed by electrophoretic methods, the pure enzyme presents structural heterogeneities even when recovered from crystals. Up to three enzyme populations could be identified by polyacrylamide gel electrophoresis and more than ten by isoelectric focusing. They have similar molecular masses and mainly differ in their charge. All are fully active. This microheterogeneity is also revealed by ion-exchange chromatography and chromatofocusing. Several levels of heterogeneity have been defined. A first type, which is reversible, is linked to redox effects and/or to conformational states of the protein. A second one, revealed by immunological methods, is generated by partial and differential proteolysis occurring during enzyme purification from yeast cells harvested in growth phase. As demonstrated by end-group analysis, the fragmentation concerns exclusively the N-terminal end of the enzyme. The main cleavage points are Gln-19, Val-20 and Gly-26. Six minor cuts are observed between positions 14 and 33. The present data are discussed in the perspective of the crystallographic studies on aspartyl-tRNA synthetase.

Amino Acid Sequence↗

Yeast tRNA(Asp)-aspartyl-tRNA synthetase complex: low resolution crystal structure.

Yeast aspartyl-tRNA synthetase, a dimer of molecular weight 125,000, and two molecules of its cognate tRNA (Mr = 24160) cocrystallize in the cubic space group I432 (a = 354 A). The crystal structure was solved to low resolution using neutron and X-ray diffraction data. Neutron single crystal diffraction data were collected in five solvents differing by their D2O content in order to use the contrast variation method to distinguish between the protein and tRNA. The synthetase was first located at 40 A resolution using the 65% D2O neutron data (tRNA matched) tRNA molecules were found at 20 A resolution using both neutron and X-ray data. The resulting model was refined against 10 A resolution X-ray data, using density modification and least-squares refinement of the tRNA positions. The crystal structure solved without a priori phase knowledge, was confirmed later by isomorphous replacement. The molecular model of the complex is in good agreement with results obtained in solution by probing the protected part of the tRNA by chemical reagents.

Amino Acyl-tRNA Synthetases↗

Comparative efficacy and safety of ceftizoxime, cefotaxime and latamoxef in the treatment of bacterial pneumonia in high risk patients.

One hundred and thirty-five patients with bacterial pneumonia who had risk factors (alcoholism, chronic obstructive pulmonary disease, corticosteroid therapy diabetes mellitus, advanced age, solid tumours) were randomly allocated in a double-blind fashion to receive either ceftizoxime (2-4 g every 8 h), cefotaxime (1-2 g every 4 h), or latamoxef (2-4 g every 8 h). Of the 84 patients evaluable for efficacy, clinical cure was achieved in 91%, 85%, and 89% of ceftizoxime- (20/22), cefotaxime-(23/27), and latamoxef-treated (31/35) patients, respectively. Adverse reactions occurred in one of 45 ceftizoxime-treated patients, one of 43 cefotaxime-treated patients, and seven of 47 latamoxef-treated patients. Abnormal laboratory values during therapy were seen in 50% of latamoxef-treated and 43% of cefotaxime-treated patients and in 29% of ceftizoxime-treated patients. Hypoprothrombinaemia occurred in five latamoxef-treated patients and one of these patients experienced an episode of haematemesis. In this study, ceftizoxime, cefotaxime, and latamoxef were similarly effective; however, the incidence of side effects was most frequent with latamoxef.

Aged↗

Analysis of the cytosolic proteins of chick embryo gonads by two-dimensional gel electrophoresis.

Gonads and mesonephros dissected from normal chick embryos, as well as a blood sample, were labelled in vitro with [35S]methionine and [14C]leucine. The patterns of cytosolic protein synthesis of the different tissues were analysed using two-dimensional (2-D) polyacrylamide gel electrophoresis. Three developmental stages, i.e. 8, 12 and 17 days of incubation, were investigated. Five sex-specific proteins were detected in the male and two in the female. In the testis, only one protein is already present at the 8-day stage. In the ovary, one protein exists since the 8-day stage, but it is also synthesized in the mesonephros. The second ovary-specific protein appeared only at the 12-day stage.

Animals↗

Case report: prosthetic valve endocarditis due to a nutritionally variant streptococcus.

The first report of prosthetic valve endocarditis due to a nutritionally variant streptococcus is presented. A 21-year-old woman developed persistent fever within one week of mitral valve replacement. Prosthetic valve dysfunction developed necessitating valve replacement. Cultures of blood and the excised prosthetic valve using routine media were negative; Streptococcus mitior grew as satellite colonies around Staphylococcus aureus and in beef heart infusion broth supplemented with 0.001% pyridoxine HCl. Treatment with penicillin G and streptomycin for six weeks was curative. Nutritionally variant streptococci should be considered in patients with prosthetic valve endocarditis and negative routine cultures.

Adolescent↗

Yeast tRNAAsp: codon and wobble codon-anticodon interactions. A transferred nuclear Overhauser enhancement study.

The conformations of the ribotrinucleoside bisphosphates GpApC and GpApU, the codon and wobble codon for aspartic acid respectively, bound to yeast tRNAAsp in solution, have been examined by means of time-dependent transferred nuclear Overhauser enhancement measurements to determine distances between bound ligand protons. The conformations of the two bound ribotrinucleoside bisphosphates are shown to be very similar with an overall root-mean-square difference in interproton distances of 0.03 nm. The ribose conformations of all the residues are 3'-endo; the glycosidic bond torsion angles of the A and C residues of GpApC and of the A and U residues of GpApU are in the low anti range. These features are typical of an A-RNA type structure. In contrast, the G residue of both GpApC and GpApU exists as a mixture of syn and anti conformations. The overall conformation of the two bound ribotrinucleoside bisphosphates is also similar to A-RNA and the stability of the complexes is enhanced by extensive base-base stacking interactions. In addition, it is shown that the binding of the codon GpApC to tRNAAsp induces self-association into a multicomplex system consisting of four GpApC-tRNAAsp complexes, whereas the wobble codon GpApU fails to induce any observable self-association.

Anticodon↗

Crystallization of transfer ribonucleic acids.

A compilation of crystallization experiments of tRNAs published in literature as well as original results are given and discussed in this paper. Up to now 17 different tRNA species originating from Escherichia coli and from the yeast Saccharomyces cerevisiae have been crystallized. All structural tRNA families are represented, namely the tRNAs with large or small extra-loops and among them the initiator tRNAs. The tRNAs with small variable loops (4 to 5 nucleotides), e.g. tRNAAsp and tRNAPhe, yield the best diffracting crystals. Crystalline polymorphism is a common feature; about 100 different crystal forms have been observed, but only 6 among them enabled structure determination studies by X-ray diffraction. Crystallization strongly depends upon experimental parameters such as the presence of polyamines and magnesium as well as upon the purity and the molecular integrity of the tRNAs. Crystals are usually obtained by vapour diffusion methods using salts (e.g. ammonium sulfate), organic solvents (e.g. isopropanol, dioxane or 2-methyl-2,4-pentane diol) or polyethylene glycol as precipitants. A methodological strategy for crystallyzing new tRNA species is described.

Crystallization↗

Branhamella catarrhalis pneumonia and immunoglobulin abnormalities: a new association.

A case of Branhamella catarrhalis pneumonia in a patient with multiple myeloma is reported. With the inclusion of this case, 5 of 17 (29.4%) reported cases of Branhamella pneumonia or empyema have occurred in patients with underlying diseases associated with immunoglobulin abnormalities. This strong clinical association suggests that qualitatively and quantitatively normal immunoglobulins are important host defense mechanisms in preventing infection with this pathogen. The sputum Gram stain demonstrating kidney-shaped gram-negative diplococci may be an early clue to the diagnosis, as well as an initial guide to empiric therapy, and may help the laboratory isolate and identify this pathogen, which, because of its morphologic resemblance to Neisseria, is frequently reported as "normal flora".

Adult↗

Large scale purification and structural properties of yeast aspartyl-tRNA synthetase.

A large scale purification procedure of baker's yeast aspartyl-tRNA synthetase is described which yields more than 200 mg pure protein starting from 30 Kg of wet commercial cells. The synthetase is an alpha 2 dimer of Mr = 125,000 +/- 5,000 which can be crystallized (J. Mol. Biol. 138, 1980, 129-135). The enzyme has an elongated shape with a Stokes radius of 50 A and a frictional ratio of 1.5. The synthetase has a tendency to aggregate but methods are described where this effect is overcome.

Amino Acyl-tRNA Synthetases↗

Crystallization of a tRNA . aminoacyl-tRNA synthetase complex. Characterization and first crystallographic data.

A complex formed between the dimeric aspartyl-tRNA synthetase from yeast (Mr congruent to 125,000) and two molecules of its cognate yeast tRNAAsp (Mr = 24,160) was crystallized using ammonium sulfate as the precipitant. The crucial parameter which governs a successful crystallization is the enzyme tRNA stoichiometry. Crystals are only obtained when the starting solution precisely contains two tRNA molecules for one enzyme molecule. It was demonstrated by electrophoresis, biological activity assays, and crystallographic data that the crystals contain the two components in the same two to one stoichiometric ratio. The crystals, of cubic shape with edges up to 0.8 mm, belong to space group 1432. The cell parameter is 354 A and the asymmetric unit contains one particle of complex. The solvent content is about 78%, higher than the values commonly observed. Although particularly soft, the quality of the crystals is suitable for x-ray diffraction studies up to 7-A resolution.

Amino Acyl-tRNA Synthetases↗

Isoelectric focusing and isoelectric points of aminoacyl-tRNA synthetases.

Isoelectric points and isoelectric focusing behaviour of 10 highly purified eukaryotic aminoacyl-tRNA synthetases from 3 sources, Saccharomyces cerevisiae, Euglena gracilis and Phaseolus vulgaris were examined. The pI-values measured on polyacrylamide gels under native conditions are situated between pH 5.0-7.5. A microheterogeneity was observed for 9 enzymes appearing otherwise homogeneous on gel electrophoresis. A compilation of the isoelectric points of aminoacyl-tRNA synthetases is given and literature data are compared with our experimental results.

Amino Acyl-tRNA Synthetases↗