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Sensitivity of group A streptococci to antibiotics. Prevalence of resistance to erythromycin in Japan.

More than 60% of all strains of group A streptococci isolated during the period from 1974 to 1975 from children with streptococcal infections in Hokkaido district, Japan, were highly resistant to erythromycin. These strains were found to be multiply resistant to lincomycin hydrochloride monohydrate, chloramphenicol, and tetracycline, and were exclusively type 12 by T-protein typing. The clinical symptoms produced by these organisms were rather mild, responded to penicillin well, and were rarely complicated with glomerulonephritis. The high prevalence of resistant group A streptococci was nationwide, which may have been related to recent excessive use of erythromycin and other macrolide antibiotics. Erythromycin can no longer be considered the drug of choice in the management of streptococcal infections in Japan. This suggests that a periodic surveillance of antibiotic sensitivity of streptococcal isolates may be necessary in other countries in which macrolide antibiotics are frequently prescribed.

Anti-Bacterial Agents↗

Unusual multiresistant Staphylococcus aureus in a newborn nursery.

An apparently unique strain of Staphylococcus aureus, resistant to penicillins (including penicillinase-resistant penicillins), cephalosporins, cephamycins, aminoglycosides (all but amikacin), tetracycline, lincomycin, erythromycin, and, in a lesser percentage, to chloramphenicol as well, was isolated on more than 100 occasions over a 20-month period in a general hospital in San Sebastian, Spain. Forty-two (60%) of the 70 patients infected were patients in the newborn nursery. Many of these infants contracted serious disease. In the epidemiologic study of the nursery, this Staphylococcus strain was found in the air and on the clothing of the staff, although no nasal, perineal, or axillary carriers were found among the personnel.

Anti-Bacterial Agents↗

A case of pneumococcal typhlitis.

A 29-year-old man had abdominal pain for 24 hours. This and the results of an abdominal examination were typical of acute appendicitis. He had suffered sinusitis for two weeks. At operation, the appendix was normal; there was an abscess in the cecal wall, the exudate of which grew pneumococci. Incidental appendectomy was done and the patient was treated successfully with lincomycin hydrochloride, and later, cephalexin monohydrate. It is possible that the typhlitis was secondary to the upper respiratory infection.

Adult↗

Drug-induced colitis as a surgical disease.

Colitis has been reported after parenteral administration of penicillin, ampicillin, streptomycin, tetracycline, and, most recently, lincomycin and its analogue, clindamycin. An incidence of diarrhea and accompanying proctocolitis as high as 20% with or without pseudomembrane formation has been noted. In most cases, withdrawal of the drug, supportive measures, and administration of corticosteroids have resulted in reversal of the colonic disease. In the past year, however, it has become apparent that there is a substantial mortality (as high as 38%) associated with the conservative management of this entity. We have used subtotal colectomy as a life-sparing procedure, and we report here its application in two specific instances, along with the clinical course, roentgenographic findings, and laboratory data. We believe drug-induced colitis is increasing in frequency and severity and is of importance to the surgeon.

Adrenal Cortex Hormones↗

Influence of environmental chemicals on drug therapy in humans: studies with contraceptive steroids.

The effects have been studied of various environmental factors on the variability in response to oral contraceptive steroid therapy in women. Ten- to thirty-fold variations in plasma concentrations of norethisterone, L-norgestrel and ethinyloestradiol have been shown in samples taken 12 h after administration of oral contraceptives in mid-menstrual cycle. Factors shown to be responsible for this variation include passage into the enterohepatic circulation, a variable first-pass effect, and changes in metabolism in the gut wall or liver due to diet, disease, smoking or administration of drugs. Phenobarbitone and the antibiotic rifampicin increase both oestrogen and progestogen metabolism in women and in experimental animals by increasing hepatic and gut wall metabolism. In animals, other antibiotics (ampicillin, neomycin and lincomycin) suppress the gut flora that normally hydrolyse steroid conjugates excreted in bile; enterohepatic circulation or oral contraceptive steroids is thus reduced and their plasma concentrations lowered by up to 90%. In the human, ampicillin has a variable but less dramatic effect on elimination of oral contraceptives. Samples of gut wall mucosa obtained from patients with coeliac disease are defective in their ability to metabolize oral contraceptives. Cigarette smokers eliminate ethinyloestradiol more rapidly than non-smokers; an increased production of reactive steroid metabolites may thus be a cause of vascular disease in women who smoke and take contraceptive steroids.

Animals↗

[Chemotherapeutic mixtures for the selective growth of anaerobes].

In order to find a selective medium for the cultivation of Gram-positive spore-less rods several chemotherapeutics have been tested. By adding of 5 micrograms metronidazol/ml to brain-heart-infusion-supplemented agar as well as by putting metronidazol discs on the agar it became possible to cultivate selective test tribes. Lincomycin (5 micrograms/ml), clindamycin (5 micrograms/ml) and novobiocin (5 micrograms/ml) inhibited almost all the test tribes. Spectinomycin (5 micrograms/ml) and neomycin (5 and 10 micrograms/ml) failed to inhibit. With 100 micrograms neomycin/ml Gram-positive anaerobes were clearly inhibited.

Anti-Bacterial Agents↗

Liquid chromatography of clindamycin 2-phosphate on triethylaminoethyl cellulose.

The separation of clindamycin 2-phosphate from clindamycin 3-phosphate, clindamycin 4-phosphate, clindamycin B 2-phosphate, and lincomycin 2-phosphate was achieved by liquid chromatography on triethylaminoethyl cellulose using a 254-nm monitor. The compounds have low molar absorptivities at 254 nm (smaller than 17), and UV detection is made possible by the high capacity support triethylaminoethyl cellulose. Linear peak height response versus concentration allows rapid quantitation of clindamycin 2-phosphate.

Borates↗

High-throughput screening for multi-class veterinary drug residues in animal muscle using liquid chromatography/tandem mass spectrometry with on-line solid-phase extraction.

A rapid qualitative method using on-line column-switching liquid chromatography/tandem mass spectrometry (LC/MS/MS) was developed and validated for screening 13 target veterinary drugs: four macrolides - erythromycin A, josamycin (leucomycin A3), kitasamycin (leucomycin A5), and tylosin A; six (fluoro)quinolones - ciprofloxacin, danofloxacin, enrofloxacin, flumequine, oxolinic acid, and sarafloxacin; and lincomycin, virginiamycin M1, and trimethoprim in different animal muscles. Clindamycin, norfloxacin, nalidixic acid, oleandomycin, ormetoprim, and roxithromycin were used as the internal standards. After simple deproteination and analyte extraction of muscle samples using acetonitrile, the supernatant was subjected to on-line cleanup and direct analysis by LC/MS/MS. On-line cleanup with an extraction cartridge packed with hydrophilic-hydrophobic polymer sorbent followed by fast LC using a short C18 column resulted in a total analysis cycle of 6 min for 19 drugs. This screening method considerably reduced the time and the cost for the quantitative and confirmatory analyses. The application of a control point approach was also introduced and explained.

Animals↗

Over-production of the D1:2 protein makes Synechococcus cells more tolerant to photoinhibition of photosystem II.

Over-expression of the psbAIII gene encoding for the D1 protein (form II; D1:2) of the photosystem II reaction centre in the Synechococcus sp. PCC 7942 was studied using a tac promoter and the lacIQ system. Over-expression was induced with 40 microgram/ml IPTG in the growth medium for either 6 or 12 h at growth irradiance (50 mumol photons m-2 s-1). This treatment doubled the amount of psbAII/III mRNA and the D1:2 protein in membranes but decreased the amount of psbAI messages and the D1:1 protein. The total amount of both heterodimeric reaction centre proteins, D1 and D2, remained constant under growth light conditions, indicating that the number of PSII centres in the membranes was not affected, only the form of the D1 protein was changed from D1:1 to D1:2 in most centres. When the cells were photoinhibited either at 500 or 1000 mumol photons m-2 s-1, in the presence or absence of the protein synthesis inhibitor lincomycin, the D1:2 protein remained at a higher level in cells in which over-expression had been induced by IPTG. These cells were also less prone to photoinhibition of PSII. It is suggested that the tolerance of cells to photoinhibition increases when most PSII reaction centres contain the D1:2 protein at the beginning of high irradiance. This tolerance is further strengthened by maintaining psbAIII gene over-expression during the photoinhibitory treatment.

Amino Acid Sequence↗

Mechanisms of macrolide resistance in Ureaplasma urealyticum: a study on collection and clinical strains.

Ureaplasma urealyticum is considered as a species which is intrinsically sensitive to macrolides (MIC less than 1 microgram/ml). Nevertheless, some of the strains recently isolated in our laboratories showed moderate to high levels of resistance (MICs ranging from 2 micrograms/ml to 100 micrograms/ml). In particular, a strain (CT28) isolated from a patient with nongonococcal urethritis long treated with erythromycin revealed a MIC greater than 100 micrograms/ml for this antibiotic. In order to investigate the mechanisms of resistance, strain CT28 and ten clinical and laboratory U. urealyticum strains were compared for the sensitivity to six antibiotics including three macrolides. Moreover the amount of macrolide uptake and the specific antibiotic binding to ribosomes were studied. Strain CT28 was resistant to josamycin, erythromycin, roxithromycin, lincomycin and clindamycin but sensitive to minocycline. When compared to a sensitive strain, strain CT28 showed a six-fold reduction in intracellular macrolide influx and accumulation and a reduction in antibiotic binding to ribosomes. The mechanisms implicated in these differences may be important for macrolide resistance in U. urealyticum.

Biological Transport↗

Molecular cloning in streptococci: physical mapping of the vehicle plasmid pSM10 and demonstration of intergroup DNA transfer.

The streptococcal resistance plasmid pSM10 (8.3 kb), a deletion derivative of pSM10419 (22.9 kb) determining constitutive erythromycin and lincomycin resistance, was physically mapped with the restriction endonucleases AvaI, AvaII, EcoRI, HpaI, KpnI, PvuII (one site each), HindIII, HaeII (three sites each), HincII (four sites), and HhaI (five sites). Using the cryptic plasmid pVA318 as cloning vehicle, the largest HindIII fragment of pSM10 (3.3 kb) was shown to contain the erythromycin/lincomycin resistance gene(s) of the plasmid. The AvaII site of pSM10 proved to be suitable as a site for cloning AvaII-generated chromosomal DNA fragments from a group C streptococcal strain in the Challis strain of Streptococcus sanguis (group) H). A detailed physical map of the chimeric plasmid pSM10221 (12.8 kb), a fusion product of pSM10 and the staphylococcal chloramphenicol resistance plasmid pC221 (4.5 kb), is also presented. The plasmid chimera has properties making it potentially useful in development of a doubly selective streptococcal cloning vehicle by searching for insertional inactivation.

Chromosome Mapping↗

Cloning and expression of a tylosin resistance gene from a tylosin-producing strain of Streptomyces fradiae.

A gene conferring high-level resistance to tylosin in Streptomyces lividans and Streptomyces griseofuscus was cloned from a tylosin-producing strain of Streptomyces fradiae. The tylosin-resistance (Tylr) gene (tlrA) was isolated on five overlapping DNA fragments which contained a common 2.6 Kb KpnI fragment. The KpnI fragment contained all of the information required for the expression of the Tylr phenotype in S. lividans and S. griseofuscus. Southern hybridization indicated that the sequence conferring tylosin resistance was present on the same 5 kb SalI fragment in genomic DNA from S. fradiae and several tylosin-sensitive (Tyls) mutants. The cloned tlrA gene failed to restore tylosin resistance in two Tyls mutants derived by protoplast formation and regeneration, and it restored partial resistance in a Tyls mutant obtained by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) mutagenesis. The tlrA gene conferred resistance to tylosin, carbomycin, niddamycin, vernamycin-B and, to some degree, lincomycin in S. griseofuscus, but it had no effect on sensitivity to streptomycin or spectinomycin, suggesting that the cloned gene is an MLS (macrolide, lincosamide, streptogramin-B)-resistance gene. Twenty-eight kb of S. fradiae DNA surrounding the tlrA gene was isolated from a genomic library in bacteriophage lambda Charon 4. Introduction of these DNA sequence into S. fradiae mutants blocked at different steps in tylosin biosynthesis failed to restore tylosin production, suggesting that the cloned Tylr gene is not closely linked to tylosin biosynthetic genes.

Cloning, Molecular↗

Isolation of a DNA sequence related to several plasmids from Bacillus thuringiensis after a mating involving the Streptococcus faecalis plasmid pAM beta 1.

The transmissible plasmid pAM beta 1, which codes for resistance to erythromycin and lincomycin, was transferred from Streptococcus faecalis to several strains of Bacillus thuringiensis by a filter-mating process. Introduction of pAM beta 1 into the Emr transconjugant strains of B. thuringiensis was confirmed by Southern hybridisation using the 32P-labelled pAM beta 1 as a probe. In the B. thuringiensis transconjugant strains, used as donors, the beta plasmid conserved its ability to be transferred during intraspecific mating, with a frequency of 10(-4) per recipient cell. In addition, the transconjugant clones acted as donors of the erythromycin resistance marker and permitted the transfer of cryptic plasmids present in the B. thuringiensis (beta) strains used as donors. From a transconjugant clone of B. thuringiensis a hybrid plasmid resulting from an in vivo insertion into pAM beta 1 of a 3 Md DNA sequence was isolated. This 3 Md DNA molecule originated from a 54 Md plasmid of a kurstaki strain and is related to several plasmids found in different serotypes of B. thuringiensis.

Bacillus thuringiensis↗

The antimicrobial susceptibility of Staphylococcus species isolated from canine dermatitis.

In a retrospective study, 1538 strains of beta-haemolysin-producing Staphylococcus species isolated from dermatitis in dogs at three veterinary clinical microbiology laboratories in Norway during 1986-87 and 1993-94 were investigated for their antimicrobial susceptibility. None of the strains was resistant to cloxacillin, cephalexin or the quinolones enrofloxacin and ciprofloxacin. More than 96% of the strains were susceptible to trimethoprim-sulphonamide, bacitracin and fucidic acid. Between 67% and 89% of the strains were susceptible to erythromycin, lincosamides, tetracycline, neomycin and chloramphenicol. Only 37.9% of the strains were susceptible to penicillin. The frequency of penicillin resistance increased significantly between the first and second periods, from 46.0% to 58.6%. The frequency of resistance to lincomycin, clindamycin and erythromycin also increased significantly between the first and second periods, from 3.0%, 2.1% and 3.3% to 25.5%, 19.5% and 24.8%, respectively. A moderate increase in resistance to tetracycline was also noted, from 20.4% in the first to 27.6% in the second period. On the other hand, the frequency of resistance to trimethoprim-sulphonamide decreased significantly from 4.1% in the first to 0.9% in the second period. Many different resistance patterns were observed in each period. However, the proportion of multiresistant strains increased from 2.1% in the first to 10.2% in the second period. There was a decrease in resistance to the combination of trimethoprim-sulphonamide and penicillin from the first to the second period. Resistance to the combination of lincosamides and penicillin increased. For the combinations penicillin-tetracycline-lincosamides, penicillin-lincosamides-erythromycin, and penicillin-tetracycline-lincosamides-erythromycin, there was a striking increase in resistance between the first and the second periods.

Animals↗

Blue light-dependent regulation of cytoplasmic ribosomal RNA synthesis in Chlorella.

Effect of blue and red light on ribosomal RNA synthesis in autotrophic synchronous cultures of Chlorella pyrenoidosa (strain 211-8b) is studied by pulse labeling experiments with tritiated guanosine. Nucleic acids were separated by electrophoresis on polyacrylamide gels. Compared with darkness and red light (679 nm), blue light (457 nm) of equal quantum flux (0.5-5x10(-10) Einstein cm-2 s-1) stimulates incorporation into ribosomal RNA. This blue light effect is observed in the cytoplasmic ribosomal RNA after 5 min of illumination, whereas the stimulation of chloroplast ribosomal RNA synthesis by blue light appears later. Maturation of chloroplast ribosomal RNA is slower than that of cytoplasmic ribosomal RNA. The blue light effect on the cytoplasmic ribosomal RNA formation does not require chloroplast RNA or protein synthesis as shown by inhibitor studies with rifampicin or lincomycin. The blocking of cytoplasmic protein synthesis by cycloheximide inhibits the blue light effect on ribosomal RNA formation. It is concluded that the cytoplasmic ribosomal RNA transcription is controlled by a blue light sensitive system.

Chlorella↗

The effects of mucus on the binding of cationized ferritin by human and animal gastrointestinal epithelium.

Human gallbladder and gastric epithelial cells are normally covered with a layer of mucus. When specimens were exposed to cationized ferritin (CF) in vitro, they did not regularly bind nor internalise it. If the tissues were first exposed to the mucolytic agents cysteamine or pepsin, then the gallbladder epithelium readily bound CF and the gastric epithelium irregularly. The in vivo binding of CF by guinea pig gallbladder could be abolished by the induction of mucous hypersecretion by the antibiotic lincomycin. The removal of the mucus by mucolytic agents restored the binding of CF. The irregular binding of CF by gastric mucosa after the use of mucolytic agents suggests other factors may be at play.

Animals↗

Drug utilization in paediatrics: non-medical factors affecting decision making by prescribers.

The study was done to show that in certain areas of paediatric pharmacotherapy unexpected discrepancies may arise between accepted therapeutic principles and the actual behaviour of a prescribing doctor. The first example was of a great reduction in penicillin use in a university teaching hospital after certain therapeutic accidents: in one year, there were 2 fatal cases of rhabdomyolysis due to use of procaine benzyl-penicillin. Other antimicrobial drugs inferior to penicillin, such as lincomycin and sulphonamides, replaced penicillins. The second example showed the inverse relationship between the use of antitussives and other drugs in symptomatic treatment of respiratory diseases in outpatients and inpatients; the pressure of unduly optimistic expectations of therapy imposes a high prescribing rate of these drugs in the outpatient population, in contrast to hospitalized patients, whose doctors, being spared such pressure, prescribe antitussives far less often. The third example demonstrates the possibility of inadequate education in the use of antimicrobial drugs. Although doctors from regional hospitals receive their training at an university hospital, they tend to prescribe chloramphenicol ten times more per bed-day than their colleagues in an university hospital. In terms of the cost/effectiveness ratio, a high prescribing rate of cephalosporins is not economically favourable in a university teaching hospital. It is also shown that studies of drug utilization in children are feasible if age--appropriate adaptation of the statistical value expressed as the defined daily dose is performed. The adaptation was evaluated by comparing pharmacy-based drug consumption data expressed in "paediatric defined daily doses", with actual days of treatment with particular drugs, i.e. data from patient records for 244 beds in the University Teaching Hospital.

Ambulatory Care↗

[Antibiotic-associated diarrhoea and enterocolitis (author's transl)].

Many antibiotics, particularly the lincomycins, may cause diarrhoea with or without enterocolitis. The pathogenesis of antibiotic-associated diarrhoea without colitis is uncertain; colloidosmotic water binding in the colon by endogenous glycoproteins undegraded by colonic bacteria is considered. Antibiotic-associated enterocolitis is now known to be due to toxin-producing clostridia, proven for Cl. difficile. Improved methods for the detection of toxin and clostridia are presently being studied. Endoscopically, pseudomembranes are characteristic but not antibiotic-specific, they may be absent or missed diagnostically. A possible role of asymptomatic clostridia-carriers in enterocolitis clustering remains to be determined. The potentially lethal course of the disease requires rapid diagnosis and therapy, with discontinuation of the antibiotic, intensive supportive measures and, at least in severe disease, oral vancomycin.

Adult↗