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Biomedical subjects

M Varon

Publications and source records attributed to M Varon.

At least 19 recordsLinked to original sources

Enzyme immunoassay for the diagnosis of cat-scratch disease defined by polymerase chain reaction.

Whole-cell immunofluorescent antibody (IFA) tests for detection of anti-Bartonella henselae immunoglobulin (Ig) G are commonly used to diagnose cat-scratch disease (CSD). The need to cultivate B. henselae in Vero cells for antigen preparation and the absence of routinely applied IFA assays for IgM constitute the major disadvantages of this form of test. We describe the results of an enzyme immunoassay (EIA) for IgM and IgG that used N-lauroyl-sarcosine-insoluble outer membrane antigens from agar-grown B. henselae performed in 84 patients with definite CSD (regional lymphadenitis, cat contact, and > or =1 confirmatory test: polymerase chain reaction, skin test, or B. henselae culture). Although this method has been used as a diagnostic tool in several case reports, it has not previously been evaluated in a large study of definitively proven CSD cases. Results of this study indicate that the EIA described herein can play an important role in the serodiagnosis of CSD, although improvement of the sensitivity, particularly that of the IgM, would be desirable.

Adolescent↗

DNA amplification for the diagnosis of cat-scratch disease in small-quantity clinical specimens.

Diagnosis of cat-scratch disease (CSD) by polymerase chain reaction (PCR) of lymph node fineneedle aspiration (FNA) and primary lesion specimens can be difficult owing to the minute amount of available material. A PCR assay specifically suited to test these specimens was developed. First, small-quantity (10 microL) samples were prepared from 17 CSD-positive and 16 CSD-negative specimens, and DNA extraction and amplification from these samples were compared using 3 methods. Sensitivity and specificity of PCR were 100% using material collected on glass microscope slides and by using Qiagen (Hilden, Germany) columns for DNA extraction. Then, this method was used to test 11 archival glass microscope slides of FNA (7 malignant neoplasms, 4 undiagnosed lymphadenitis) and 2 primary lesion specimens. Two of the 4 lymphadenitis samples and the 2 primary lesion specimens were PCR positive. The technique presented could facilitate CSD diagnosis from a wider range of clinical samples.

Bartonella henselae↗

Organization and cell-cell interaction in starved Saccharomyces cerevisiae colonies.

Cell growth in yeast colonies is a complex process, the control of which is largely unknown. Here we present scanning electron micrographs of Saccharomyces cerevisiae colonies, showing changes in the pattern of cell organization and cell-cell interactions during colony development. In young colonies (</=36 h), cell density is relatively low, and the cells seem to divide in a random orientation. However, as the colonies age, cell density increases and the cells seem to be oriented in a more orderly fashion. Unexpectedly, cells in starved colonies form connecting fibrils. A single connecting fibril 180 +/- 50 nm wide is observed between any two neighboring cells, and the fibrils appear to form a global network. The results suggest a novel type of communication between cells within a colony that may contribute to the ability of the community to cope with starvation.

Saccharomyces cerevisiae↗

Rpb7 can interact with RNA polymerase II and support transcription during some stresses independently of Rpb4.

Rpb4 and Rpb7 are two yeast RNA polymerase II (Pol II) subunits whose mechanistic roles have recently started to be deciphered. Although previous data suggest that Rpb7 can stably interact with Pol II only as a heterodimer with Rpb4, RPB7 is essential for viability, whereas RPB4 is essential only during some stress conditions. To resolve this discrepancy and to gain a better understanding of the mode of action of Rpb4, we took advantage of the inability of cells lacking RPB4 (rpb4Delta, containing Pol IIDelta4) to grow above 30 degrees C and screened for genes whose overexpression could suppress this defect. We thus discovered that overexpression of RPB7 could suppress the inability of rpb4Delta cells to grow at 34 degrees C (a relatively mild temperature stress) but not at higher temperatures. Overexpression of RPB7 could also partially suppress the cold sensitivity of rpb4Delta strains and fully suppress their inability to survive a long starvation period (stationary phase). Notably, however, overexpression of RPB4 could not override the requirement for RPB7. Consistent with the growth phenotype, overexpression of RPB7 could suppress the transcriptional defect characteristic of rpb4Delta cells during the mild, but not during a more severe, heat shock. We also demonstrated, through two reciprocal coimmunoprecipitation experiments, a stable interaction of the overproduced Rpb7 with Pol IIDelta4. Nevertheless, fewer Rpb7 molecules interacted with Pol IIDelta4 than with wild-type Pol II. Thus, a major role of Rpb4 is to augment the interaction of Rpb7 with Pol II. We suggest that Pol IIDelta4 contains a small amount of Rpb7 that is sufficient to support transcription only under nonstress conditions. When RPB7 is overexpressed, more Rpb7 assembles with Pol IIDelta4, enough to permit appropriate transcription also under some stress conditions.

Dimerization↗

Trans-acting regulation of antibiotic TA genes in Myxococcus xanthus.

Two regulatory mutations of Myxococcus xanthus, which cause an increase in the transcription of genes required for antibiotic TA synthesis, were mapped by transduction and their effect on transcription of four TA genes examined. The two regulatory mutations were closely linked and located within the 40-kb TA gene cluster on the M. xanthus chromosome. Recombinants were constructed which contained one of the regulatory mutations and promoter probes in the four different TA genes. Both regulatory mutations enhanced transcription of three of the four TA genes. However, construction of a strain containing the over-expression regulatory mutation in a wild-type background produced less antibiotic than the parental strain.

Anti-Bacterial Agents↗

Restraining patients as part of hospital policy.

In Israel, senior citizens spend more time in hospitals than do members of any other age group. Chronic, age-related illnesses make the senior population especially dependent on the hospital system. Hospitalization, however, has its own risks-frequent and/or prolonged hospital stay can increase susceptibility to infections and complications, and can lead to impairment of mobility and of overall functioning. In this study, we'll be taking a look at the use of patient restraints in geriatric institutions. We'll look at the factors that induce attending staff to restrain elderly patients, the kinds of restraints used, and the feeling of the staff on this controversial issue. We'll also present the results of interviews with nursing services administrators in several geriatric institutions on the subject of restraining policies and procedures. Our findings indicate that procedures and policies regarding restraint vary from one institution to another. In some cases there are no clear written procedures, and patients are restrained without the written and signed order of a physician. We also found that attending staff are not sufficiently knowledgeable about the legal issues involved in patient restraint.

Aged↗

Mutation and mapping of genes involved in production of the antibiotic TA in Myxococcus xanthus.

Transposition of TnV and Tn5lac into Myxococcus xanthus yielded 8,381 kanamycin-resistant mutants that were tested for antibiotic TA production. Twenty-four of the mutants were nonproducers of TA (less than 0.4 ng/ml), and 3 produced a higher level (2.5 micrograms/ml) than the parent strain (1.5 micrograms/ml). For most of the strains, there was 100% cotransduction between kanamycin resistance and the altered TA phenotype. Southern blot analysis of restriction digests of the mutant DNA indicated that the transposons were inserted at different sites on the M. xanthus chromosome. The TA genes were mapped by cotransduction between pairs of mutants following replacement of the initial insert of one of the pair with the tetracycline resistance transposon Tn5-132. Nine of the 13 nonproducers tested were linked over a 36-kb stretch of the chromosome. There was no linkage between one of the overproducers and any of the nonproducers tested.

Anti-Bacterial Agents↗

Use of Tn5lac to study expression of genes required for production of the antibiotic TA.

The beta-galactosidase activities arising from Tn5lac insertions in several genes required for antibiotic TA production were measured under different growth conditions. In all of the non-TA-producing mutants, the beta-galactosidase specific activity was higher when the cells were grown in nutrient-limited 0.5CTS medium (0.5% Casitone plus alanine, serine, and glucose) than in rich 2CT medium (2% Casitone). One of the mutants, 420, had low beta-galactosidase specific activity in both media. The other seven mutants containing inserts in genes essential for TA production had specific activities of 139 to 367 U/mg of protein in 0.5CTS medium and 11 to 48 U/mg of protein in 2CT medium. The beta-galactosidase specific activities of two strains, 1030 and 420, increased during exponential growth in 0.5CTS medium. The beta-galactosidase specific activities of both strains increased greatly when the cells were grown in the presence of magnesium phosphate, which traps ammonium ions. The Tn5lac insertions in 1030 and 420 were used to screen for mutants with increased levels of transcription. An N-methyl-N'-nitro-N-nitrosoguanidine-induced mutation in 1030 that mapped 17 kb from the omega 1010 insert increased the specific activity of beta-galactosidase 21 times in 2CT medium. The regulatory mutation appears to release the repression caused by 2CT medium. A UV-induced mutation in 420 increased the beta-galactosidase specific activity 1.4 to 2.4 times. Medium conditions that affect the transcription of TA genes are discussed in terms of enhanced antibiotic TA production.

Anti-Bacterial Agents↗

Effect of adhesive antibiotic TA on plaque and gingivitis in man.

The adhesive antibiotic TA was applied to the dento-gingival junction of 8 human volunteers, suffering from moderate to severe gingivitis. 2 diametrically opposed quadrants of the mouth received 4 applications of 0.1 mg TA, while the other 2 quadrants were treated with a placebo and served as controls. The plaque index, gingival index and bleeding index were scored periodically for 2 weeks and in 4 patients for up to 30 days. The TA-treated quadrants showed a rapid decrease in all 3 indices following 2 treatments. A further improvement was observed with the 3rd and 4th treatments. 30 days after the onset of the experiment and 23 days after the last application, the indices were still considerably lower than the initial values.

Adult↗

Cell-density-dependent killing of Myxococcus xanthus by autocide AMV.

Autocide AMV of Myxococcus xanthus was purified and identified as phosphatidylethanolamine. Alkaline hydrolysis of AMV yielded a high proportion of mono- and diunsaturated fatty acids. The bactericidal activity of AMV on M. xanthus depended upon the density of target cells: the greater the cell density, the greater the killing by AMV. For example, at 2 U of AMV per ml, 0, 50, and 99% killing was measured with 2 X 10(4), 2 X 10(5), and 2 X 10(7) target cells per ml, respectively. The cell-density-dependent activity of AMV was also observed on solid medium. Studies with model lipid compounds suggest that the inhibitory activity of AMV is due to the fatty acid moiety, released from phosphatidylethanolamine by the concerted (enzymatic) activity of many cells. Mutants of M. xanthus selected for resistance to AMI (a mixture of fatty acids) were also resistant to AMV. The possible role of AMV in developmental lysis is discussed.

Chromatography, Thin Layer↗

Gaucher's disease associated with chronic lymphocytic leukaemia.

The association of Gaucher's disease with chronic lymphocytic leukaemia (CLL) is very rare. To our knowledge only five cases have been reported (Chang-Lo, Yam & Rubenstone 1967; Chang-Lo et al. 1975; Fox et al. 1984; Lee 1982; Mark, Dominguez & Rywlin 1982). We report a patient with this association, the CLL had been diagnosed 12 years after the demonstration of Gaucher's disease.

Bone Marrow Cells↗

Myxococcus xanthus autocide AMI.

Autocide AMI of Myxococcus xanthus was purified and shown to be a mixture of fatty acids: 46.4% saturated, 49.3% monounsaturated, and 4.3% diunsaturated. The specific autocidal activities (units per milligram) were as follows: purified AMI, 1,000; saturated fraction, 100; monounsaturated fraction, 800; diunsaturated fraction, 2,200. Model fatty acids mimicked to some extent the activity of AMI, although none of the fatty acids tested were as active as purified AMI. Spontaneous and induced mutants of M. xanthus were selected for resistance to AMI and to fatty acids. The AMI-resistant mutants were also resistant to the model fatty acids, whereas resistance to fatty acids was specific to the compound used for mutant selection. All AMI- and fatty acid-resistant mutants examined were found to be blocked in fruiting body formation. Some of these mutants were able to form normal fruiting bodies when mixed with the extracellular fluid of the parental strain. The data suggest that AMI plays a role in developmental lysis of M. xanthus.

Bacteriolysis↗

Adsorption of antibiotic TA to dental hard tissues.

Antibiotic TA (TA) is a wide-spectrum, bactericidal antibiotic produced by Myxococcus xanthus strain TA. It was previously demonstrated that TA binds tightly to soft tissues while retaining its bactericidal activity in the bound form. The present study was undertaken to investigate TA adsorption to dental hard tissues. Slabs of dental tissues that had been cut from periodontally-involved extracted human teeth were treated with TA and then washed in saline with shaking (saline being replaced every 15 minutes). After 30, 60, and 120 minutes of washing, 45, 39, and 27% of the input TA activities were retained on the slabs, respectively. The tooth-bound TA was released slowly into the aqueous medium in active form. Similar experiments with beta-lactam antibiotics resulted in no significant adhesion to the slabs. The study demonstrates that TA binds tightly to dental tissues while retaining its bactericidal properties.

Adsorption↗

Autocides produced by Myxococcus xanthus.

Ethanol extracts of Myxococcus xanthus contained several substances, referred to as autocides, which were bactericidal to the producing strain but showed no activity against other bacteria. The autocides were produced by growing cells and remained largely cell bound throughout the growth cycle; ca. 5% of the autocidal activity was found in the supernatant fluid at the time cell lysis began. The autocides were separated by sequential-column and thin-layer chromatography into five active fractions (AM I through AM V). Each of the fractions was at least 20 times more active against M. xanthus than against the other gram-negative or gram-positive bacteria tested. AM I, AM IV, and AM V were inactive against yeasts, whereas a mixture of fractions AM II and AM III was active against Rhodotorula sp. At low concentrations, AM I reversibly inhibited the growth of M. xanthus; at higher concentrations of AM I, the cells lysed within 1 h. The lowest concentration of AM IV that showed any activity caused rapid cell death and lysis. The mode of action of the major autocide, AM V, was different from that of AM I and AM IV. During the initial 2 h of treatment, the viable count of M. xanthus cells remained constant; during the next few hours killing occurred without lysis; within 24 h lysis was complete. The autocidal activity of each of the fractions was expressed when the cells were suspended in buffer, as well as in growth medium. The possible role of autocides in developmental lysis of M. xanthus is discussed.

Anti-Bacterial Agents↗

Multisystem presentation of eosinophilic fasciitis.

A 20-year-old man experienced the onset of progressive scleroderma-like skin changes with eosinophilia and hypergammaglobulinaemia after strenuous military activity. A biopsy showed faciitis. Concomitant splenomegaly, polysynovitis, restrictive lung function, myositis, decrease hepatic clotting factors and proteinuria were documented. He responded well to corticosteroids, relapse upon their discontinuation and had a complete remission of both skin and systemic features while on penicillamine and azathioprine. He is new in good health and has received no medication for the past two years.

Adult↗