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A simple method for differentiation of Propionibacterium acnes and Propionibacterium propionicum.

TLC glycolipid profiles of several culture collection and clinical strains of Propionibacterium acnes and Propionibacterium propionicum were examined. The former were characterized by weak orcinol-positive minor glycolipids of type g, while the others had mainly strong orcinol-positive major glycolipids of type G. The simple and rapid small scale procedure seemed to be useful for differentiation of these phenotypically similar and genotypically closely related species irrespective of their serotypes.

Bacterial Typing Techniques

Complement activation in acne vulgaris: in vitro studies with Propionibacterium acnes and Propionibacterium granulosum.

To better define the role of bacteria in inflammatory acne vulgaris, we have investigated the ability of four strains of Propionibacterium acnes and three strains of Propionibacterium granulosum to activate complement. Complement activation was assayed by incubating normal human serum with varying concentrations of each strain and measuring residual total hemolytic complement activity. When serum was tested unaltered, P. acnes strains were approximately threefold more potent than an equal weight of P. granulosum in consuming complement, which could reflect classical and/or alternative pathway activation. All strains also consumed complement in serum chelated with ethyleneglycol-bis (beta-aminoethyl ether)-N,N'-tetraacetic acid, which selectively assays alternative pathway activation. Incubation of unaltered serum with both P. acnes and P. granulosum resulted in immunoelectrophoretic conversion of C4, C3, and factor B of the alternative pathway. Incubation of chelated serum resulted in conversion of C3 and factor B. These data taken together suggest that both species can activate complement through either pathway. Serum incubated with P. acnes was chemotactic for polymorphonuclear leukocytes, and this chemotactic activity was largely C5 dependent as shown by antibody inhibition. It is suggested that complement activation may occur in vivo in acne, and the inflammatory response may be contributed to by the generation of C5-dependent chemotactic factors.

Acne Vulgaris

Comparative studies of porphyrin production in Propionibacterium acnes and Propionibacterium granulosum.

Porphyrin production by Propionibacterium acnes and that by Propionibacterium granulosum were compared. Porphyrin synthesized by both organisms was identified as coproporphyrin III on the bases of absorption and fluorescence spectra and behavior on paper chromatography and thin-layer chromatography. Quantitative, rather than qualitative, differences in production were found between these organisms. In general, P. granulosum produced significantly greater amounts (P less than 0.001) of porphyrin than did P. acnes. delta-Aminolevulinic acid synthetase appeared to be the rate-limiting enzyme of the heme biosynthetic pathway in both organisms. The increased porphyrin production in P. granulosum is apparently associated with increased delta-aminolevulinic acid synthetase activity.

5-Aminolevulinate Synthetase

Esterification of (4--14C)cholesterol by cutaneous bacteria (Staphylococcus epidermidis, Propionibacterium acnes, and Propionibacterium granulosum).

Cholesterol and cholesteryl esters constitute 3 to 5% of the lipids derived from human sebaceous glands. The present study demonstrates that cutaneous bacteria, Staphylococcus epidermidis, and to a lesser extent Propionibacterium acnes, and Propionibacterium granulosum, esterify [4--14C]cholesterol to steryl esters in vitro. The degree of esterification is a characteristic of the strain of organism, ranging from 0 to 32.7% in the organisms tested. Cholesterol esterification in these experiments was dependent on the addition of homogenates of isolated, pooled sebaceous glands to the incubation medium. Indirect evidence suggests that the fatty acids from sebaceous triglycerides are utilized for cholesterol esterification by the bacteria. These results indicate that the proportion of steryl esters in skin surface lipids may be influenced by the density of the microbial population in the skin.

Carbon Radioisotopes

Biosynthesis of vitamin B12. Some properties of the 5,6-dimethylbenzimidazole-forming system of Propionibacterium freudenreichii and Propionibacterium shermanii.

1. Homogenates of Propionibacterium freudenreichii transform riboflavin into 5,6-dimethylbenzimidazole. This process is stimulated by nicotinamide. Homogenates of Propionibacterium shermanii form only small amounts of 5,6-dimethylbenzimidazole from riboflavin in the absence of nicotinamide, but also form appreciable amounts in the presence of nicotinamide. 2. The stimulation of the 5,6-dimethylbenzimidazole-forming system by nicotinamide shows a lag phase which is abolished by preincubation of the homogenate with nicotinamide. Since no lag phase is observed when nicotinamide is replaced by nicotinate, nicotinate seems to be the true stimulating agent. These observations are in agreement with the fact that nicotinamide is rapidly split to nicotinate in homogenates of P. freudenreichii. 3. The 5,6-dimethylbenzimidazole-forming homogenate system is only active at a high buffer concentration (0.3--0.5 M) and in the presence of oxygen. The system has a pronounced oxygen optimum. 4. Flavin mononucleotide and flavin-adenine dinucleotide are better substrates for the 5,6-dimethylbenzimidazole-forming homogenate system than riboflavin. But with [1'-14C]riboflavin as substrate the specific radioactivity of 5,6-dimethylbenzimidazole is higher than the specific radioactivity of flavin--adenine dinucleotide and lower than the specific radioactivie substrate for the formation of 5,6-dimethylbenzimidazole. 5. A tentative reaction sequence for the transformation of flavin mononucleotide into 5,6-dimethylbenzimidazole is discussed.

Benzimidazoles

Cytotaxin production by comedonal bacteria (Propionibacterium acnes, Propionibacterium granulosum and Staphylococcus epidermidis).

The potential role of different species of comedonal bacteria as chemotactic stimuli in the inflammatory phase of acne vulgaris was investigated by comparing 12 strains of Staphylococcus epidermidis, 11 strains of Propionibacterium acnes, and 5 strains of P. granulosum for production of cytotaxin in vitro. Results indicated that not only were there marked differences in cytotaxin production between different strains of the same species grown under identical growth conditons, but there were often significant differences in cytotaxin activity of the same strain grown in different media. This finding is discussed in relation to development of inflammation in quiescent comedones in acne vulgaris.

Acne Vulgaris

The functioning of cytochrome b in the electron transport to furmarate in Propionibacterium freudenreichii and Propionibacterium pentosaceum.

Fumarase-free electron particles from Propionibacterium freudenreichii and P. pentosaceum were prepared by discontinuous sucrose gradient centrifugation, and the influence of 2-n-heptyl-4-hydroxy-quinoline-N-oxide (HQNO) and ultraviolet irradiation on the reduction of menaquinone and cytochrome b with L-lactate or glycerol-3-phosphate and the reoxidation by fumarate was studied. In the presence of HQNO the steady state reduction level of menaquinone during fumarate reduction was increased whereas the steady state reduction level of cytochrome b was decreased as compared with the reduction levels measured in the absence of HQNO. The steady state reduction level of menaquinone during electron transport to fumarate was not influenced by ultraviolet irradiation and the steady state reduction level of cytochrome b was decreased at increasing irradiation times. The data indicate that cytochrome b is involved in the electron transport to fumarate.

Aerobiosis

Hepatoprotective effect of SY-640, a novel acetamide derivative, on Propionibacterium acnes and lipopolysaccharide-induced liver injury in mice.

The hepatoprotective effect of SY-640 on Propionibacterium acnes and lipopolysaccharide-induced liver injury in mice and its protective mechanism were examined. Oral administration of SY-640, 150 mg/kg once daily for 7 days, significantly inhibited Propionibacterium acnes and lipopolysaccharide-induced liver injury, but a single administration was without effect. Liver-infiltrating cells (T-lymphocytes and macrophages) play an important role in Propionibacterium and lipopolysaccharide-induced liver injury and express a higher level of leukocyte function-associated antigen-1. SY-640 inhibited the number of liver-infiltrating cells and attenuated the increased expression of leukocyte function-associated antigen-1 on these cells. Tumor necrosis factor-alpha mediated Propionibacterium acnes and lipopolysaccharide-induced liver injury, and SY-640 inhibited the elevation of the serum tumor necrosis factor-alpha concentration after injection of lipopolysaccharide in Propionibacterium acnes-primed mice. The putative effects of SY-640 are inhibitory effects on infiltration into the liver and on activation of T-lymphocytes and macrophages after Propionibacterium acnes-priming, and attenuation of expression of cell adhesion molecules such as leukocyte function-associated antigen-1. The immunological effect of SY-640 is likely to be closely related to the inhibition of Propionibacterium and lipopolysaccharide-induced liver injury.

Acetamides

Cloning, sequencing, and expression of pyrophosphate-dependent phosphofructokinase from Propionibacterium freudenreichii.

Pyrophosphate-dependent 6-phosphofructo-1-kinase (PPi-PFK) from Propionibacterium freudenreichii is a non-allosteric enzyme with properties dissimilar to those of other described phosphofructokinases. The enzyme was cloned into pBluescript, sequenced, and expressed in Escherichia coli at levels 15 times higher than those observed in Propionibacterium. The gene consists of 1215 bases which code for a protein of 404 amino acids and a mass of 43,243 daltons. High G + C in the codon usage (66%) of the gene is consistent with the classification of Propionibacterium in the High-G + C subdivision of the Gram-positive bacteria. While showing no sequence identity to the non-allosteric ATP-dependent phosphofructokinase of E. coli, alignments of the amino acid sequence with other PFKs reveal degrees of identities among the amino halves of the proteins, from 26% between the Propionibacterium and potato PPi-PFKs, and 29% between Propionibacterium and E. coli ATP-PFKs. These levels of identities indicate that the amino halves of these proteins are homologous. Identities between the carboxyl half of Propionibacterium PFK and carboxyl halves of other sequences are below 20%, suggesting that the carboxyl half is not homologous. Despite the poor conservation, most of the residues that take part in the binding of fructose-6-P or Mg-PPi could be readily identified by analogy to the structure of the E. coli PFK. Both the fructose-6-P and ATP-binding sites are conserved, indicating that PPi binds to the homologous site of the E. coli ATP-binding site.

Adenosine Triphosphate

Infection caused by Propionibacterium in children.

Three hundred eighty-six isolates of Propionibacterium sp. were recovered from 2,003 specimens studied for the identification of anaerobic bacteria in children during a 15-year period. Three hundred forty-three (89%) of these were Propionibacterium acnes. A total of 50 (13%) Propionibacterium isolates identified from 45 patients were considered to cause infection. Clinically significant infections caused by Propionibacterium sp. were associated with bacteremia in 10 children; ear infection in eight; abscesses in seven; adenitis and central nervous system infection in five each; burns in four; wounds in three; tumors and bone in two each; and cysts, eye, sinus, and mastoid in one each. Predisposing or underlying conditions were present in 33 children (73%). These included the presence of a foreign body (17), immunodeficiency (six), malignancy (five), diabetes (five), previous surgery (four), and steroid therapy (two). Antimicrobial therapy was given to 41 (91%) children. Surgical drainage was concomitantly performed in 22 (49%). Four patients died. This study highlights the importance of Propionibacterium sp. as an unusual, but potentially important, pathogen in children.

Child

Kinetics of nitric oxide synthase induction by Propionibacterium avidum and lipopolysaccharide.

Conditions for the induction of rat liver Ca2(+)-independent nitric oxide synthase were determined with killed Propionibacterium avidum, and compared with lipopolysaccharide endotoxin. Similar maximal induction was obtained intraperitoneally with the two types of inducers but killed Propionibacterium avidum gave a long-lasting induction while lipopolysaccharide displayed a rapid and short response. Moreover, the induction resulting from an intravenous administration of killed Propionibacterium avidum reached 60 times that of the control whereas lipopolysaccharide treatment induced a 24-fold stimulation only. It is noteworthy that with the first inducer the nitric oxide activity was stable with time whereas with the second one it dropped after 8 h. Whatever the route of administration of killed Propionibacterium avidum, some huge vacuolated Kupffer cells were found in the liver whose parenchyma was almost normal. Numerous monocytes, and unaltered Kupffer cells, were observed. Kupffer cells were identified to be responsible for the uptake of killed Propionibacterium avidum.

Animals

Infections caused by Propionibacterium species.

Eight hundred and sixteen isolates of Propionibacterium species (725 of which were Propionibacterium acnes) were isolated from 3,971 specimens submitted for the identification of anaerobic bacteria over the course of 10 years. A total of 94 Propionibacterium isolates (12%) identified in 92 patients were considered to cause infection. The rest of the isolates were determined to be contaminants or of uncertain pathogenic significance. Significant infections caused by Propionibacterium species were associated with the blood in 15 patients, central nervous system in 11, lymph glands in 10, abscesses in eight, joints in seven, wounds in seven, cysts in six, and sinuses in five. Predisposing or underlying conditions were noted in 66 patients (70%). The most common conditions were the presence of foreign bodies (29 patients), diabetes (12), previous surgery (10), trauma (seven), malignancy (seven), immunodeficiency (seven), and steroid therapy (four). Antimicrobial therapy was administered to 83 patients; for 47 patients this therapy was given in conjunction with surgical drainage or correction. Surgical drainage alone was performed in nine patients. Five patients (5%) died. These data illustrate that although Propionibacterium species are rarely associated with infections, these organisms can occasionally cause serious infections.

Adolescent

Transfer of Propionibacterium innocuum Pitcher and Collins 1991 to Propioniferax gen. nov. as Propioniferax innocua comb. nov.

16S ribosomal DNA analysis indicates that Propionibacterium innocuum is a phylogenetic neighbor of Luteococcus japonicus and that this pair of organisms branches intermediately between the genus Propionibacterium on the one side and the genera Aeromicrobium and Nocardioides on the other side. Phenotypically, strains of P. innocuum differ from species of Propionibacterium by exhibiting aerobic growth and possessing arabinose in the cell wall, they differ from species of Aeromicrobium and Nocardioides by the formation of propionic acid, and they differ from species of Luteococcus in morphology. Consequently, P. innocuum should not be classified with authentic Propionibacterium species, and the transfer of P. innocuum Pitcher and Collins 1991 to a new genus, Propioniferax, as Propioniferax innocua gen. nov., comb. nov. is proposed.

DNA, Bacterial

Mode of protection of mice against herpes simplex virus type 2 infection by Propionibacterium.

We compared various strains of Propionibacterium with regard to protection of young adult mice against lethal infection with herpes simplex virus type 2 (HSV-2). Propionibacterium acnes, P. granulosum, and P. avidum were protective, while P. acidi-propionici and P. lymphophilum were ineffective. The protective effect proved to be in the cell wall fraction. Attempts were made to elucidate possible mechanisms of the protection using both effective and ineffective strains. The results strongly suggest that induction of interferon rather than activation of macrophages and natural killer cells by Propionibacterium pretreatment plays a crucial role, directly or indirectly, in protection against infection by herpes simplex virus. Propionibacterium only moderately protected newborn mice against HSV-2 infection.

Adjuvants, Immunologic

Identification of Actinomyces, Arachnia, Bacterionema, Rothia, and Propionibacterium species by defined immunofluorescence.

Fractionated fluorescein-isothiocyanate (FITC)-conjugated immunoglobulin G (dye-to-protein ratio <10), produced against whole cells of Actinomyces spp., Arachnia, Bacterionena, Rothia, and Propionibacterium spp., give species-specific conjugates with controlled nonspecific staining reactions when appropriately diluted on the basis of their antibody content (10 mg/ml). Using this standardization in immunofluorescence, serotype-specific conjugates are also available after dilution for all serotypes of these organisms except for Actinomyces viscosus type 2, and Propionibacterium acnes type 1. Adequately adsorbed conjugates could be used to differentiate these serotypes from A. viscosus type 1 and P. acnes type 2, respectively. A serological classification in defined immunofluorescence corresponded to species and serotype designation proposed on the basis of other serological analysis and biochemical characteristics. This includes a separation in immunofluorescence of two serotypes of Propionibacterium acnes. The detection of certain actinomycetes of the family Actinomycetaceae and Propionibacterium species by the defined immunofluorescence in direct smears prepared from clinical specimens agreed to 88% with parallel culturing when including a prereduced (PRAS) medium technique for isolation. Qualitative studies revealed that single cells of these organisms could be specifically identified by immunofluorescence when admixed with morphologically similar bacteria and a large number of other contaminants.

Actinomyces

[Propionibacterium acnes in the etiology of endocarditis].

Propionibacterium acnes is the gram positive anaerobic bacteria belongs to the normal skin and oral microbial flora. The participation of this microorganism in the infective endocarditis is still controversial. The aim of the study was to perform the diagnostic and therapeutic difficulties in 5 patients with infective endocarditis caused by Propionibacterium acnes. In 3 out of 5 patients the infective endocarditis developed after prosthesis valve replacement, in 2 others on the native valves. The inserted prostheses were mechanical ones, propionibacterium acnes was identified as causative organisms in all of the causes (two positive blood and/or valve culture). The bacterial strains were sensitive to the antibiotics as: penicillins, cephalosporins, clindamycin, and vancomycin, however cephalosporins used at the beginning of the treatment in 3 patients and clindamycin in 1 patient had limited clinical efficacy. Later treatment with timentin, augmentin and tienamycin was successful in 3 patients; one patient was cured with vancomycin. One patient died because of septic, embolic complication in early stage of illness. We conclude the effectiveness of penicillins in combination with clavulanic acid and tienamycin in therapy of infective endocarditis due to Propionibacterium acnes. The treatment should be lasted during 4-6 weeks.

Adult

The correlation of susceptibility of different Propionibacterium strains to macrophage killing and antitumor activity.

The stimulation of the reticuloendothelial system and antitumor activity against sarcoma-L-1 of three different strains of Propionibacterium was investigated. The bacteria injected intraperitoneally with a single dose of 1 mg per mouse appeared to possess different effectiveness in both assays used. The same strains were examined for their susceptibility to phagocytosis and intracellular killing by murine peritoneal macrophages. The results obtained showed correlations between Propionibacterium resistance to degradation by phagocytic cells and their antitumor and stimulatory activities. It is concluded that the potency of antitumor and stimulatory activities of different strains of Propionibacterium are closely related to their cell wall structure.

Animals

Enrichment of Propionibacterium in paddy soil by addition of various organic substances.

Enrichment of Propionibacterium and production of propionic acid in paddy soil after addition of glucose, casamino acids, or Na-lactate, respectively, were investigated. Only in the case of Na-lactate, both enrichment of Propionibacterium (74% of total anaerobes isolated) and production of large amounts of propionic acid were observed. These results suggest that lactate may support the growth of Propionibacterium in paddy soil.

Amino Acids