Molecular basis of streptomycin resistance in Mycobacterium tuberculosis: alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosomal RNA pseudoknot

Multidrug-resistant strains of Mycobacterium tuberculosis have resulted in several recent outbreaks. Recognition of drug resistance is important both for treatment and to prevent further transmission. Here we use molecular biology techniques to study the basis of streptomycin resistance in single and multi-drug-resistant M. tuberculosis. We demonstrate that streptomycin resistance is associated with mutations implicated in ribosomal resistance. The mutations found either lead to amino acid changes in ribosomal protein SI2 or alter the primary structure of the 16S rRNA. The 16S rRNA region mutated perturbs a pseudoknot structure in a region which has been linked to ribosomal S12 protein.     

Confirmation of a dopamine metabolite in parkinsonian brain tissue by gas chromatography—mass spectrometry

Gas chromatography—mass spectrometry was used to identify a dopamine metabolite isolated from the substantia nigra of parkinsonian brain tissue. Incubation of dopamine with monoamine oxidase B gave the same product which was identified as 3,4-dihydroxyphenylacetaldehyde. The structure of the compound was established by chemical synthesis, metastable ion measurement and high-resolution mass spectrometry.     

STUDIES OF VEGETATIVE COMPATIBILITY-INCOMPATIBILITY IN HIGHER PLANTS. IV. THE DEVELOPMENT OF TENSILE STRENGTH IN A COMPATIBLE AND AN INCOMPATIBLE GRAFT

Three phases of adhesion between the stock and scion are observable during the formation of a compatible autograft in Sedum telephoides and an incompatible heterograft between Sedum telephoides and Solanum pennellii. The first phase of adhesion is similar in both systems in that it 1) lasts 2 to 3 days, and 2) is characterized by an average increase in tensile strength of 1 g breaking weight (BW)/mm² graft area (GA)/day. In the compatible Sedum autograft, the second phase of adhesion lasts from Days 3 to 11 after grafting and is correlated with a 28-fold increase in the tensile strength of the graft union to approximately 56 g BW/mm² GA by 11 days after grafting. The third phase of adhesion in the compatible autograft is characterized by a leveling off of the tensile strength of the graft union at approximately 56 g BW/mm² GA, roughly equal to that of an ungrafted internode. Graft formation is now complete. These results suggest that the ratio of the tensile strength of the graft union : tensile strength of a comparable ungrafted internode provides an estimate of the percent development of compatible autografts. In the incompatible heterograft between Sedum and Solanum, Phase II adhesion 1) lasts from Days 2 to 5 after grafting, and 2) peaks at 12 g BW/mm² GA at 5 days after grafting. Phase III adhesion in the incompatible heterograft occurs subsequent to Day 5 after grafting and is characterized by an average decrease in the tensile strength of the graft union of 0.3 g BW/mm² GA/day. The results of this study are discussed relative to the quantitative contributions of various structural events to the tensile strength of a graft union.     

Effect of rhesus or vervet cytomegalovirus infection of DNA synthesis in untreated and 5-iodo-2''-deoxyuridine-arrested cells.

Infection of permissive cells with either rhesus or vervet cytomegalovirus resulted in suppression of cellular DNA synthesis, only viral DNA was resolved by isopycnic centrifugation after 24 h postinoculation. Infection of 5-iodo-2'-deoxyuridine (IUdR)-arrested cells with either of the simian cytomegaloviruses, however, induced synthesis of cellular DNA of normal density; synthesis of cellular DNA substituted with IUdR as evidenced by resolution of a heavy DNA peak after isopycnic centrifugation was not observed. Stimulation of DNA synthesis in IUdR-arrested cells was not observed with virus inactivated with UV light.     

Effects of Light and Temperature on the Association between Zea mays and Spirillum lipoferum

Zea mays was grown on a low N nutrient solution under 16 conditions of light and temperature in a crossed-gradient room in an attempt to determine whether or not variation in climatic conditions influences N₂ fixation by the association between maize and Spirillum lipoferum. Temperatures were 28, 32, 36, and 40 C and 10 C lower at night; light intensities were 500, 1,250, 2,400, and 3,000 ft-c. Plants harvested after 94 days showed no significant benefit from association with S. lipoferum either in dry weight production or in total N content; variations in temperature and light had only a small influence on N₂ fixation under the conditions tested. Measurements of total N, together with designated assumptions, indicated that less than the equivalent of 0.5 kilogram of N was fixed/hectare during the entire growing period by the maize-S. lipoferum association. Rates of C₂H₂ reduction by replicate root samples generally were low and variable and did not correlate with the measurements of total N.     

The Vascularization of the Kidneys in Bufo bufo (L.), Bombina variegata (L.), Rana ridibunda (L.) and Xenopus laevis (D.) (Amphibia,Anura) as Revealed by Scanning Electron Microscopy of Vascular Corrosion Casts

Abstract The vascular patterns of the ventral side of the kidneys in Bufo bufo and Rana ridibunda is similar. Strong venulae renales revehentes dominate. In Bombina variegata and Xenopus laevis, however, also many superficially located glomeruli are found at the ventral side. In Xenopus further branching of the renal arteries into afferent arterioles attracts attention. Bundles of delicate afferent arterioles originate within circumscribed areas of the renal artery. The glomerular layer of the kidney is tightest in Bombina, and has its maximal extension in Rana. It covers up to 2/3 of the thickness of the kidney while in the other species studied the glomeruli are restricted to the ventral third of the kidney. Glomeruli with double afferent or efferent arterioles were rarely found in Xenopus. The vascularization of the dorsal side of the kidneys is characterized by the presence of large (Bufo, Rana, Xenopus) or small (Bombina) venulae renales advehentes.     

The Vascularization of the Anuran Brain: The Mesencephalon

The angioarchitecture of the toad's mesencephalon is studied by scanning electron microscopy of vascular corrosion casts. The arterial supply is performed by the superior mesencephalic artery and by an artery arising from the ramus posterior of the arteria carotis cerebralis just in front of the retroin-fundibular communicating artery. The venous drainage is exerted by the vena diencephalica, the vena lateralis mesencephalis and by anterior and posterior branches of the encephaloposthypophysial portal vein. Characteristics of the mesencephalic angioarchitecture are the centrifugal direction of the blood flow, the intensive capillarization, and the formation of vascular meshworks in the region of the stratum griseum periventriculare tecti and of the stratum griseum superficiale tecti.     

Acute and chronic infection of human lymphoblastoid cell lines with measles virus.

Several human continuous lymphoblastoid cell lines (LCL) having T or B characteristics were infected with low and high passage strains of measles virus. All of the cell lines were susceptible to one or the other or to both strains of measles virus with the production of typical syncytial giant cells and released cell-free infectious virus into the supernatant medium. There was no consistent pattern of susceptibility of LCL with either T or B characteristics to infection by measles virus. Viral induced cytolysis of the lymphoblastoid cells in many of the lines was marked, but in the LCL that could be maintained over longer periods of time, a state of chronic, less cytolytic and persistent infection could be established. The infection was characterized by the production of moderate amounts of cell-free infectious virus for up to 4 1/2 months after initial infection with little change in the number of viable cells in culture. Long-term low multiplicity of infection (MOI) experiments demonstrated that the cell-free infectious virus was being produced only by a small number of cells, but the majority of cells in culture contained measles antigen that was in a cell-restricted, noninfectious, or defective form. Electron microscopic examination of the chronically infected cells demonstrated that many of them contained aggregates of hollow tubular intranuclear nucleocapsids whose "stripped" appearance was in marked contrast to the larger granular intracytoplasmic nucleocapsids found during earlier stages of infection. It is theorized that the persistent infection of LCL may serve as a model in understanding the immune mechanisms which permit latent and chronic measles infection in man.     

Thrombin, epidermal growth factor, and phorbol myristate acetate stimulate tubulin polymerization in quiescent cells: a potential link to mitogenesis.

Previous studies suggest that alterations in the microtubule (MT)-tubulin equilibrium during G0/G1 affect mitogenesis. To determine the effect of growth factors on the MT-tubulin equilibrium, we developed a radioactive monoclonal antibody binding assay (Ball et al.: J. Cell. Biol. 103:1033-1041, 1986). With this assay, 3H-Ab 1-1.1 binding to cytoskeletons in confluent populations of cultured cells is proportional to the number of tubulin subunits polymerized into MTs. We now show that purified alpha-thrombin increases 3H-Ab 1-1.1 binding to cytoskeletons of serum-arrested mouse embryo (ME) fibroblasts from 1.5- to 3-fold. This stimulation is dose-dependent and correlates with concentrations of thrombin required for initiation of DNA synthesis. Other mitogenic factors, epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA), also stimulate MT polymerization. Addition of colchicine (0.3 microM) eight hours after growth factor addition, blocks stimulation of 3H-thymidine incorporation by thrombin, EGF, or PMA, suggesting that tubulin polymerization or subsequent events triggered by MT polymerization are required for cells to enter a proliferative cycle. Consistent with models for autoregulation of tubulin synthesis, thrombin, EGF, and PMA all increase tubulin synthesis 9 to 15 hr after growth factor addition, raising the possibility that the decrease in free tubulin and subsequent stimulation of tubulin synthesis is linked to progression of cells into a proliferative cycle. Colchicine addition to these cells also stimulates DNA synthesis, but colchicine-stimulated cells enter S phase 6 to 8 hr later than those stimulated by growth factors. This delayed stimulation may be related to the time required for degradation of tubulin-colchicine complexes below a critical level. These data suggest that regulation of cell proliferation may be linked to increased MT polymerization and the resulting decrease in free tubulin pools.