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PubMed1977

Induction of Escherichia coli and Vibrio cholerae enterotoxins by an inhibitor of protein synthesis.

Enterotoxigenic Escherichia coli or Vibrio cholerae 569B (Inaba) grown in the presence of the antibiotic lincomycin, an inhibitor of protein synthesis, produced elevated levels of heat-labile enterotoxin or choleragen, respectively, as assayed by both vascular permeability factor and capacity to elicit fluid accumulation in rabbit ileal loops. This induction of enterotoxin did not reflect either a coupling of lincomycin resistance with increased enterotoxigenicity or an effect of lincomycin on cellular release of enterotoxin, since spontaneously isolated lincomycin-resistant mutants of both E. coli and V. cholerae still required lincomycin for induction, and large increases in E. coli permeability factor activity were found intracellularly as well as extracellularly. After the period of exponential growth, E. coli became refractory to induction by lincomycin, although most of the induced enterotoxin activity appeared only after this period. No increase in copy number of the enterotoxin plasmid in E. coli 711 (P307) was found in induced cells by analysis of deoxyribonucleic acid reassociation kinetics. These and other data suggest that synthesis of enterotoxin, or at least its accumulation, is normally limited by cellular factors whose synthesis is preferentially inhibited by lincomycin. A possible connection between this phenomenon and lincomycin-associated diarrhea is considered.

Animals↗
PubMed1988

Inactivation of lincosaminide antibiotics in Staphylococcus. Identification of lincosaminide O-nucleotidyltransferases and comparison of the corresponding resistance genes.

Resistance to lincomycin by inactivation has been detected in numerous clinical isolates of Staphylococcus; in crude extracts of Staphylococcus haemolyticus BM4610 and Staphylococcus aureus BM4611, inactivation of lincomycin and clindamycin requires the presence of a nucleoside 5'-triphosphate (ATP, GTP, CTP, or UTP) as nucleotidyl donor and Mg2+ as cofactor. The biochemical mechanism of lincosaminide inactivation was elucidated by determination of the structure of inactivated lincomycin and clindamycin by physicochemical techniques, including UV absorption spectrophotometry, 31P and 1H nuclear magnetic resonance, and periodate oxidation. In the two strains, inactivation of lincomycin gave rise to lincomycin 3-(5'-adenylate), whereas clindamycin was inactivated through its conversion to clindamycin 4-(5'-adenylate). The gene linA' encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase in S. aureus BM4611 has been sequenced and displays 93% homology with the gene linA encoding the 3-lincomycin, 4-clindamycin O-nucleotidyltransferase found in S. haemolyticus BM4610. The two enzymes are 161 amino acids long and differ by 14 amino acid substitutions.

Amino Acid Sequence↗
PubMed1977

Effect of linocmycin on the chlorophyll protein complex I content and photosystem I activity of greening leaves.

1. Greening barley and pea leaves treated with lincomycin have a reduced chlorophyll content. Lincomycin dose not alter the proportion of chlorophyll in chlorophyll-protein complex II (CPII) but greatly reduces that in chlorophyll-protein complex I (CPI). 2. Difference spectra show that chloroplasts from lincomycin-treated leaves are deficient in at least two long wavelength forms of chlorophyll ratio of chloroplasts is unaffected by lincomycin but the photochemical P-700/chlorophyll ratio is less than half of that of the control. It is less affected than the chlorophyll-protein complex I content. 4. Photosystem I activity expressed on a chlorophyll basis is unaffected by linocomycin but the light intensity for half saturation is increased 8-fold. 5. Chlorophyll-protein complex I apoprotein content is reduced by lincomycin. No evidence was found for an accumulation of its precursor(s). The relative abundance of major peptides of 18 000, 15 000 and 12 000 daltons in lincomycin-treated chloroplasts is attributed to a general inhibition of greening and associated membrane of formation.

Chlorophyll↗
PubMed1983

Effect of subminimal inhibitory concentrations of antimicrobial agents on the piliation and adherence of Neisseria meningitidis.

Neisseria meningitidis is known to be highly resistant to lincomycin (minimum inhibitory concentration greater than 32 micrograms/ml). However, during studies on meningococcal piliation, we noticed a significant reduction in the number of pili after cultivation on lincomycin-containing selective media. This observation was followed up by in vitro and in vivo studies on the relation between lincomycin and meningococcal adherence to human epithelial cells. We found a remarkable decrease in in vitro piliation and adherence after exposure to lincomycin at concentrations as low as 0.05 micrograms/ml. By giving four healthy meningococcal carriers lincomycin orally for 3 to 6 days, the possible in vivo effect of lincomycin was studied. A marked decrease in the meningococcal counts of the pharyngeal secretion was observed. One person completely lost his meningococcal strain during the observation period.

Adhesiveness↗
PubMed1976

Prophylaxis against anaerobic sepsis in bowel surgery.

Sixty-two patients were admitted to a prospective randomized controlled trial to investigate the influence of a prophylactic antibiotic, lincomycin, on anaerobic sepsis following bowel surgery. The incidence of postoperative sepsis was reduced from 45 to 18 per cent (P less than 0-025). Wound infections were reduced from 38 to 12 percent (P less than 0-05). Intra-abdominal or pelvic abscess occurred in 1 of the treated group compared with 3 controls. Septicaemia occurred after operation in 1 patient receiving lincomycin and in 3 of the controls; in 2 of the latter, pure growths of bacteroides were isolated from the blood cultures and 1 of these patients died. Although lincomycin had no influence on the number of patients who developed aerobic postoperative infections, there was a significant reduction in the incidence of sepsis due to bacteroides, which occurred in 10 of the control group compared with 1 in the lincomycin group (P less than 0-005). No patients developed complications attributable to lincomycin, such as pseudomembranous colitis. These data indicate that the genus Bacteroides are important pathogenic organisms and are responsible for postoperative morbidity. Furthermore, anaerobic sepsis can be reduced by appropriate prophylactic antibiotics.

Aged↗
PubMed1982

[Cooperative functioning of 2 systems of protection against its own antibiotic in Str. erythreus 1571, a superproducer of erythromycin].

Protein synthesis in the mycelium of various ages and the protoplasts of Str. erythreus 1571 was slightly sensitive even to high concentrations of erythromycin and lincomycin. Oxytetracycline and neomycin in low concentrations induced marked inhibition of the protein synthesis. 14C-Erythromycin was binding with the mycelium of the organism producing it in much lower amounts than with the mycelium of Str. roseolus, a lincomycin-producing culture. No products of erythromycin biotransformation by the cell-free preparations of Str. erythreus containing cofactors for enzymatic phosphorylation and demethylation were detected with the methods of microbiological titration and radiochromatography. Enzymatic N-demethylation of lincomycin was observed in the cell-free preparations of Str. erythreus. Erythromycin and lincomycin had practically no effect on polylysine synthesis in the cell-free system of Str. erythreus with poly-A and inhibited the synthesis of this polypeptide in the cell-free systems of E. coli MRE 600 and B. subtilis ATCC 6633. These antibiotics had either no effect on the fragment (puromycin) reaction with the ribosomes of Str. erythreus. In case of the ribosomes of E. coli MRE 600 the fragment reaction was sensitive to lincomycin but not to erythromycin. The ribosomes isolated from Str. erythreus and their 50S subunits bound 14C-erythromycin in much lower amounts than those isolated from E. coli and their 50S subunits. The data are indicative of the presence of 2 systems protecting the culture from erythromycin in Str. erythreus 1571, i.e. at the level of ribosomes and at the level of the cytoplasmic membrane.

Biotransformation↗
PubMed1979

Resistance of Mycoplasma pneumoniae to macrolide and analogous antibiotics.

1. The susceptibility of M. pneumoniae to antibiotics can be determined by the microtiter method. The adequate technique requires that the final volume of broth medium in a well is 0.2 ml and that the dilution is made after the parent solution of antibiotic in the test tube is dropped into a well every fifth wells. 2. M. pneumoniae was cultured on agar media containing two-fold concentrations of macrolide and analogous antibiotics, and the following results were obtained. 1) The growth of eight strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of thet strains of M. pneumoniae on agar media containing two-fold concentrations of the antibiotics revealed that, in six strains, one CFU (colony forming unit) per 10(5) to 10(6) CFU of an inoculum dose was resistant to the antibiotics. 2) The MIC (minimum inhibitory concentration) of erythromycin for the subculture of the colony grown as an average of 0.5 to eight on agar media containing erythromycin in four strains was 0.1 to 1.6 micrograms/ml in some colonies, and 400 to 800 micrograms/ml in most colonies. The results disclosed that the broth culture contains a small number of mycoplasma cells with a definite, high degree of resistance to the antibiotics, but no cells with intermediate degrees of resistance. 3) The FH strain was made resistant to erythromycin, oleandomycin, midecamycin, acetylspiramycin, leucomycin, josamycin, tylosin, lincomycin, or clindamycin by subculture in broth medium from the colony grown at the highest concentrations of each of the antibiotics in agar media. The degree of the resistance developed was 16 to 128,000 in the MIC radio and showed high values of MIC in most strains. The resistance developed was not lost by subculturing the resistant strain in broth medium without antibiotic. 4) The FH strain made resistant to the antibiotics had cross resistance to other macrolides. Strains resistant to some of the antibiotics had cross resistance to lincomycin and clindamycin, and strains resistant to others did not. Some strains made resistant to macrolides with cross resistance to lincomycin and clindaymycin and strains made resistant to lincomycin or clindamycin had no cross resistance to vernamycin B alpha, while all the resistant strains without cross resistance to lincomycin and clindamycin had cross resistance to vernamycin B alpha. No strain had cross resistance to vernamycin A...

Agar↗
PubMed2001

Putative lmbI and lmbH genes form a single lmbIH ORF in Streptomyces lincolnensis type strain ATCC 25466.

The lincomycin-production gene cluster of the industrial overproduction strain Streptomyces lincolnensis 78-11 has been sequenced (Peschke et al. 1995) and twenty-seven putative open reading frames with biosynthetic or regulatory functions (lmb genes) identified. Two distinct hypothetical genes, lmbI and lmbH, were found downstream of the lmbJ gene, coding for LmbJ protein, which is believed to participate in the last lincomycin biosynthetic step, i.e. conversion of N-demethyllincomycin (NDL) to lincomycin. In the present study, we demonstrate the presence of a single larger open reading frame, called lmbIH, in the lincomycin low-production type strain Streptomyces lincolnensis ATCC 25466, instead of two smaller lmbI and lmbH genes. The product, LmbIH, is a protein of an unknown function and is homologous with the T1dD protein family. Escherichia coli T1dD protein was previously shown to be involved in the control of DNA gyrase by LetD protein. Moreover, our experiments indicate co-regulation of lmbJ and lmbIH expression. This translation coupling probably reflects an eight nucleotide overlap between the lmbJ and lmbIH genes, as well as the lack of a Shine-Dalgarno sequence upstream of the lmbIH gene.

Amino Acid Sequence↗
PubMed1975

Antibiotics in human hematoma and wound fluid.

Using the total hip replacement patient, we measured the level of antibiotic in the wound at the time of surgery, the wound drainage for 36 to 48 hours after surgery and over a 10 day period in incubated surgical debris obtained at the time of total hip replacement. The antibiotics sodium oxacillin, lincomycin hydrochloride and sodium cephalothin were administered in standard doses at 6 hour intervals. In one group of patients the antibiotic was begun at the time of surgery, in a second group the antibiotic was begun 24 hours after the completion of surgery. Sodium oxacillin, lincomycin hydrochloride and sodium cephalothin produce antibiotic levels in the venous serum consistently above the level obtained in the simultaneous would drainage specimens. Sodium oxacillin and lincomycin hydrochloride used as described in this study are capable of bathing the area of the surgical wound with concentrations of antibiotics above the minimal inhibitory concentrations. This is evidenced by the antibiotic levels above minimal inhibitory concentrations found in the wound drainage. Sodium cephalothin levels in the wound drainage were below measurable levels in 3 of 4 patients. Sodium oxacillin and lincomycin hydrochloride antibiotic levels in the surgical wound debris and simultaneous venous clotted specimens diminished progressively in a similiar manner indicating no additional effects of the surgical wound debris on the antibiotic's biologic effectiveness. Sodium cephalothin dropped below measurable levels in three days. Measurable levels of antibiotics appeared in the wound aspirate well above the minimal inhibitory concentration in patients receiving their initial dose of sodium oxacillin 24 hours after surgery.

Anti-Bacterial Agents↗
PubMed1977

Selective medium for isolation of Bacteroides nodosus.

The sensitivity of Bacteroides nodosus, the causative agent of sheep foot rot, to 24 selected antimicrobial agents was tested. Many contaminants ordinarily associated with foot rot lesions were sensitive to lincomycin, whereas B. nodosus demonstrated resistance to this antibiotic. A concentration of 1 microgram of lincomycin per ml of basal medium optimally inhibited contaminants while allowing growth of B. nodosus. The basal medium was Eugon agar with 0.2% yeast extract and 10.0% defibrinated horse blood. Parallel inoculations of 31 foot rot lesion specimens onto basal medium and basal medium containing lincomycin (selective medium) were performed. B. nodosus was isolated from 16 of the specimens cultured on the selective medium and from only 3 of the specimens cultured on the basal medium. The concentration of agar in the standard Eugon medium was found to influence the growth of B. nodosus in the presence of lincomycin. Trimethoprim also exhibited potential selectivity for B. nodosus.

Agar↗
PubMed2006

Programmed cell death in plants: effect of protein synthesis inhibitors and structural changes in pea guard cells.

Pea leaf epidermis incubated with cyanide displayed ultrastructural changes in guard cells that are typical of apoptosis. Cycloheximide, an inhibitor of cytoplasmic protein synthesis, and lincomycin, an inhibitor of protein synthesis in chloroplasts and mitochondria, produced different effects on the dynamics of programmed death of guard cells. According to light microscopy data, cycloheximide reinforced and lincomycin suppressed the CN(-)-induced destruction of cell nuclei. Lincomycin lowered the effect of cycloheximide in the light and prevented it in the dark. According to electron microscopy data, the most pronounced effects of cycloheximide in the presence of cyanide were autophagy and a lack of apoptotic condensation of nuclear chromatin, the prevention of chloroplast envelope rupturing and its invagination inside the stroma, and the appearance of particular compartments with granular inclusions in mitochondria. Lincomycin inhibited the CN(-)-induced ultrastructural changes in guard cell nuclei. The data show that programmed death of guard cells may have a combined scenario involving both apoptosis and autophagy and may depend on the action of both cytoplasm synthesized and chloroplast and mitochondrion synthesized proteins.

Apoptosis↗