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Biomedical subjects

R Laufs

Publications and source records attributed to R Laufs.

At least 163 records · Page 9Linked to original sources
PubMed1979

Molecular characterization of a small Haemophilus influenzae plasmid specifying beta-lactamase and its relationship to R factors from Neisseria gonorrhoeae.

The ampicillin-resistant Haemophilus influenzae strain Ve445 which caused purulent meningitis and septicaemia in a newborn child in Germany contained a 4.4 megadalton (Mdal) plasmid (pVe445) and produced a TEM type beta-lactamase. The transformation to ampicillin resistance of a sensitive Escherichia coli strain with isolated pVe445 DNA proved that the structural gene for the beta-lactamase resided on this plasmid genome. Molecular DNA-DNA hybridization studies and electron microscope DNA heteroduplex analysis indicated that pVe445 probably contained 38 to 41% of the ampicillin translocation DNA segment (TnA) found on R factors of enteric origin. The TnA fragment present in pVe445 most likely does not contain both of the inverted repeat sequences of TnA. DNA-DNA polynucleotide sequence studies indicated that the 4.4 Mdal plasmid pVe445 was unrelated to the 30 to 38 Mdal H. influenzae R plasmids but was closely related to the 4.1 Mdal ampicillin resistance specifying H. influenzae plasmid RSF0885 isolated in the U.S.A. The H. influenzae plasmid pVe445 shared 91% of its base sequences with the beta-lactamase specifying Neisseria gonorrhoeae plasmid pMR0360 (4.4 Mdal) and had 85% of its base sequences in common with the beta-lactamase specifying N. gonorrhoeae plasmid pMR0200 (3.2 Mdal). All of the four 3.2 to 4.4 Mdal beta-lactamase specifying R plasmids of H. influenzae and N. gonorrhoeae investigated probably have a common evolutionary origin.

Ampicillin↗
PubMed1979

Molecular nature of two Haemophilus influenzae R factors containing resistances and the multiple integration of drug resistance transposons.

The 36-megadalton Haemophilus influenzae R plasmid pHK539 was found to specify resistance to tetracycline (Tc) and ampicillin (Ap). It was shown by molecular hybridization studies and by electron microscopy that the plasmid pHK539 contained the tetracycline translocation deoxyribonucleic acid (DNA)segment (TnTc) as well as the ampicillin translocation segment (TnAp). The TnAp was integrated in the stem of TnTc. The 34-megadalton H. influenzae R plasmid pRI234 carried a translocatable DNA segment which specified both tetracycline and chloramphenicol (Cm) resistance. Self-annealing and DNA-DNA heteroduplex experiments indicated that this transposon is probably composed of TnTc containing an insertion of a chloramphenicol resistance transposon (TnCm). TnCm is inserted into one of the components of the TnTc inverted repetitions and is itself flanked on both sides by long inverted repetitions. The H. influenzae plasmids pHK539 and pRI234 had more than 60% of their polynucleotide sequences in common with all the other 30- to 40-megadalton R factors recently found in H. influenzae isolates from different countries. The tetracycline-chloramphenicol resistance transposon of pRI234 was integrated twice at different sites in the plasmid after its growth in medium containing tetracycline. The presence of the two copies of the transposon was correlated with higher minimum inhibitory concentrations against tetracycline as well as against chloramphenicol. After its growth in medium containing tetracycline, the H. influenzae R plasmid pFR16017 specifying Tc resistance contained one, two, three, or even four copies of TnTc integrated at different sites in the plasmid, or the loop of TnTc was amplified. The heterogeneity of the pFR16017 plasmid was seen in all single-colony isolates and correlated with a higher minimum inhibitory concentration against tetracycline.

Ampicillin↗
PubMed1978

[Infectious resistance to antibiotics in Haemophilus influenzae (author's transl)].

Ampicillin-resistant Haemophilus influenzae does occur now in the FRG. In one isolate a plasmid with resistance genes (R-factor) could be demonstrated as cause of the ampicillin resistance. This R-factor influences production of a beta-lactamase of the TEM type which destroys ampicillin. The infectious nature of the ampicillin resistance was proven by the fact that it was transferable to other bacterial species through cocultivation. Parallel to ampicillin resistance tetracycline resistant Haemophilus influenzae has occurred in the FRG. Here the resistance was equally bound to plasmids. These R-factors are infectious as well. Molecular analysis of the 3 isolated resistance factors in Haemophilus influenzae showed that they carry the same resistance genes which are known from R-factors of Enterobacteriaceae. In the therapy of purulent infections due to Haemophilus influenzae such as childhood meningitis one can no longer rely on general ampicillin sensitivity of the offender. Apart from ampicillin and tetracycline resistant Haemophilus influenzae chloramphenicol resistance has been observed in a few cases.

Ampicillin↗
PubMed1978

Molecular properties of transmissible R factors of Haemophilus influenzae determing tetracycline resistance.

The tetracycline-resistant Haemophilus influenzae strains LU121 and FR16017, recently isolated in West Germany, each harbour a plasmid; that of the former (pLU12U) has a mol. wt of 31.5 X 10(6) and that of the latter (pFR16017) has a mol. wt of 33 X 10(6). Conjugation and DNA-DNA hybridization studies have shown that both plasmids are self-transmissible and carry tetracycline-resistance genes. The purified plasmid DNA of H. influenzae strain LU121 transformed a sensitive Escherichia coli strain to tetracycline resistance. The two R factors are closely related to the H. influenzae plasmid specifying ampicillin resistance (pKRE5367). Electron microscope DNA heteroduplex analysis indicated that pLU121 and pFR15017 probably carry the tetracycline-resistance transposon TnD and that pKRE5367 probably carries the ampicillin-resistance transposon TnA. There is more than one integration site for the insertion which probably represents TnD in pFR15017. All three plasmids have a similar plasmid core and could have a common evolutionary origin.

DNA, Bacterial↗
PubMed1978

Antibiotic resistance factors and other plasmids in bacterial isolates from hospitalized patients.

Among 186 arbitrarily selected gram-negative as well as gram-positive bacterial strains isolated from hospitalized patients, 63% were found to harbour plasmids, 42% of the strains with extrachromosomal DNA showed more than one plasmid DNA band in the agarose gel electrophoresis. The highest plasmid frequency was found in the enteric gram-negative microorganisms: 95% to 76% of the analyzed Enterobacter species, Klebsiella pneumoniae and E. coli isolates harboured plasmids. The plasmid frequency in the Staphylococci and group D Streptococci ranged between 36% and 75%. The examined Pneumococci strains were sensitive to all antibiotics and were found to be free of plasmids. In 45% of the enteric strains studied, the resistance markers were found to be transferable to a sensitive E. coli recipient by conjugation. Some of the resistance genes had not formed a stable complex with a plasmid and were lost during in vitro passage of their bacterial host. The loss of R-factors in vitro causes false diagnostic findings in regard to the in vivo antibiotic sensitivity of such strains. With purified plasmid DNA, individually isolated from E. coli strains, transformants were obtained carrying ampicillin resistance specifying plasmids of different molecular size. This result indicates that plasmids of different molecular size in a single bacterial cell can simultaneously carry ampicillin resistance genes. All findings indicated that the plasmids play an important role for the transfer and the maintenance of antibiotic resistance specifying genes in the bacterial isolates from clinical patients. However, most of the functions coded for by the numerous plasmids in the bacterial isolates remained unknown.

Anti-Bacterial Agents↗
PubMed1978

Detection and preliminary characterization of extrachromosomal DNA in clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis.

Isolates of Staphylococcus aureus and Staphylococcus epidermidis frequently harboured extrachromosomal DNA. From the 20 Staphylococcus aureus strains examined 85% contained extrachromosomal DNA and 65% carried more than one plasmid DNA species. The results obtained with the 19 Staphylococcus epidermidis strains analyzed were similar, 84% of the strains had extrachromosomal DNA and 68% contained several DNA species. The agarose gel electrophoresis of the DNA obtained from cleared lysates of the staphylococci proved to be efficient for the detection of plasmids and the determination of their molecular weights. The gel method appeared therefore to be suitable for the establishment of the role of plasmids in the phage typability of staphylococci. Using a Staphylococcus epidermidis strain as an example it was shown by electron microscopy that the extrachromosomal DNA demonstrated by the agarose gel electrophoresis represented covalently closed circular plasmid DNA. This plasmid DNA could be sufficiently labeled with 3H-thymidine to undertake studies on the relationship and molecular characterization of staphylococcal plasmids.

DNA, Bacterial↗
PubMed1976

Oncornaviruslike particles in human malignancies.

Mesenchymal and epithelial human malignancies were examined in the simultaneous detection test for the presence of particles with 70 S RNA and reverse transcriptase. Out of the 23 tumors tested 14 were found to be positive (61%). The finding that 2 uterine cancers contained 70 S RNA and reverse transcriptase indicates that uterine cancers may belong to the list of malignancies where the involvement of oncornaviruses is suspected. Out of the 10 human sarcomas examined in this study 4 were shown to contain particles with properties of oncornaviruses.

Centrifugation, Density Gradient↗
PubMed1976

Vaccination of nonhuman primates with killed oncogenic herpesviruses.

Killed herpesvirus cancer vaccines were prepared by inactivation of oncogenic herpesviruses (Herpesvirus saimiri and Herpesvirus ateles) with heat and formaldehyde. The killed vaccines, which are not free of viral nucleic acid, were safe in 121 vaccinated monkeys of 4 different species. Some of these monkeys have now been under observation for 2 years. The vaccines induced high titers of specific humoral antibodies in all vaccinated monkeys. The killed vaccines proved to be efficient against the i.m. challenge with cell-free oncogenic herpesviruses. The challenged animals remained clinically well without signs of an infection and have now been under observation for 1 year, while the nonvaccinated control monkeys died of malignant lymphoma 34 to 52 days after inoculation. The protection against H. saimiri could be passively transmitted with the serum of the vaccinated monkeys. The vaccination did not prevent but delayed tumor development after tumor cell transplantation. The resistance against the oropharyngeal route of infection remains to be determined.

Animals↗
PubMed1975

The effect of an immunosuppressive drug (azathioprine) on the latent infection of marmoset monkeys (Callithrix jacchus) with the oncogenic Herpesvirus saimiri.

Callithrix jacchus (CJ) marmoset monkeys can be latently infected with Herpesvirus saimiri (HVS). In order to determine whether this resistance to the oncogenic potential of HVS could be due to the immune surveillance, azathioprine, a known immunosuppressive drug, was given to 7 latently infected CJ marmosets. The animals died within 147 days probably from side effect caused by azathioprine, but no animal developed a tumor.

Animals↗
PubMed1975

A killed vaccine derived from the oncogenic Herpesvirus ateles.

A killed herpesvirus vaccine was prepared by inactivation of the oncogenic Herpesvirus ateles (HVA) with heat and formaldehyde. The vaccine proved safe in 9 marmosets (Saguinus oedipus) during a period of 461 days. The vaccinated marmosets developed high titers of serum antibodies against HVA and were resistant to 316 lethal dose 50 (LD50) cell-free HVA; the nonvaccinated control monkeys died of malignant lymphoma. The 5 challenged monkeys are clinically well and have now been under observation for 362 days.

Animals↗
PubMed1975

No evidence for particles encapsulating RNA-instructed DNA polymerase and high molecular weight virus-related RNA in herpesvirus induced tumours of non-human primates.

The simultaneous detection test gave no evidence for the presence of RNA tumour viruses in herpesvirus induced malignant lymphomas of non-human primates. The 12 tumours tested were obtained from three different monkey species inoculated with Herpesvirus saimiri or herpesvirus ateles. Particles encapsulating RNA-instructed DNA polymerase and high mol. wt. virus-related RNA were easily demonstrated in tumours of the mouse induced by type-C or type-B oncornaviruses and in human lymphoid cells infected with simian sarcoma virus type I which were examined in parallel. Attempts to demonstrate partial expression of an oncornavirus genome in the herpesvirus induced tumours and attempts to detect an interspecies antigen related to monkey oncornaviruses were negative and strengthened the observations made with the simultaneous detection test.

Animals↗
PubMed1975

Experiments to vaccinate marmoset monkeys against malignant lymphoma.

Humoral antibodies can be induced in cotton topped (CT) marmosets with killed Herpesvirus saimiri (HVS) vaccines. The serum antibodies delayed, but did not prevent, malignant lymphoma in actively and passively immunized monkeys after inoculation with 10(3.8)TCID50 of cell-free HVS. Experiments are in progress to determine whether the vaccines are able to protect against smaller challenge dosages of HVS. The immunized monkeys were not resistant against transplantation of tumor cells. Infection with Epstein-Barr virus (EBV) and Marek's disease virus (MDV) did not protect against the tumors caused by superinfection with HVS. Attenuation of HVS was not achieved by passaging the HVS genome through different monkey species.

Animals↗