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Esterase zymograms of Proteus and Providencia.

The intracellular esterases of 80 strains of Proteus and Providencia were analysed by the acrylamide-agarose zymogram technique using several synthetic substrates. The esterase bands were classified in five main groups. The alphaA-esterase bands hydrolysed alpha-naphthyl acetate and were resistant or relatively insensitive to di-isofluoropropyl phosphate (DFP). The alphaB-esterase band hydrolysed both alpha-naphthyl acetate and alpha-naphthyl butyrate and were very sensitive to DFP. Both groups of esterase bands were inactivated by heat. The betaA- and betaB-esterase bands hydrolysed beta-naphthyl acetate and were sensitive to DFP; these were distinguishable by the difference in their relative activity towards beta-naphthyl butyrate and in their relative stability to heat. The alpha-beta-esterase bands hydrolysed alpha- and beta-naphthyl acetates and alpha- and beta-naphthyl butyrates; they were inactivated by heat and were sensitive to DFP. The distribution of these esterase bands among the strains of Proteus and Providencia and their electrophoretic patterns established esterase profile types which correlate with the classification based on traditional bacteriological tests. The degree of inter-strain similarity in esterase pattern varied highly among species. The homogeneity of Proteus mirabilis and especially of Providencia stuartii contrasted with the heterogeneity of other species. This disparity suggests that the bacteria of the tribe Proteae have not the same degree of intra-specific differentiation in physico-chemical properties of esterases.

Butyrates↗

The association of particular types of Proteus with chronic suppurative otitis media.

During a period of 12 months, 57 strains of Proteus were isolated from the ears of 38 unrelated patients with chronic suppurative otitis media. Each strain was identified, typed for bacteriocin production and sensitivity, and tested for Dienes compatibility. The majority of the strains (79%) were P. mirabilis; all but one of the remainder were P.vulgaris. Although 19 different bacteriocin production/sensitivity types were found, two rare types, P. mirabilis P7/S5,12 and P. vulgaris P0/S9, were associated with 47% of these infections. This was confirmed by Dienes typing. Patients with bilateral ear disease carried a different strain in each ear. There was no evidence that persistence of infection had arisen because of the development of antibiotic resistance. Although there was some evidence that persistence may have been the result of reinfection, the isolation of these rare types of Proteus from so many patients with chronic suppurative otitis media may indicate that they play an important role in the pathogenesis of the disease. Most of the Proteus isolates were of "non-faecal" types and it is believed that infection had arisen by a route other than the faecal-aural one.

Adolescent↗

Crystallization of urine mineral components may depend on the chemical nature of Proteus endotoxin polysaccharides.

Formation of infectious urinary calculi is the most common complication accompanying urinary tract infections by members of the genus Proteus. The major factor involved in stone formation is the urease produced by these bacteria, which causes local supersaturation and crystallization of magnesium and calcium phosphates as carbonate apatite [Ca(10)(PO(4))(6).CO(3)] and struvite (MgNH(4)PO(4).6H(2)O), respectively. This effect may also be enhanced by bacterial polysaccharides. Macromolecules of such kind contain negatively charged residues that are able to bind Ca(2+) and Mg(2+), leading to the accumulation of these ions around bacterial cells and acceleration of the crystallization process. The levels of Ca(2+) and Mg(2+) ions bound by whole Proteus cells were measured, as well as the chemical nature of isolated LPS polysaccharides, and the intensity of the in vitro crystallization process was compared in a synthetic urine. The results suggest that the sugar composition of Proteus LPS may either enhance or inhibit the crystallization of struvite and apatite, depending on its chemical structure and ability to bind cations. This points to the increased importance of endotoxin in urinary tract infections.

Crystallization↗

Abolition of swarming of Proteus by p-nitrophenyl glycerin: general properties.

Para-nitrophenyl glycerin (PNPG) was shown to be an effective agent to abolish the swarming of Proteus mirabilis and Proteus vulgaris on predried solid culture media. The level required to abolish swarming varied with the strain of Proteus, the components of the medium, and also with the conditions of incubation. Generally 0.1 to 0.2 mM PNPG effectively abolished swarming for at least 24 h with aerobic incubation. Levels of PNPG that abolished swarming showed no effect upon the growth of the cells, little or no effect upon the motility characteristics of the organisms, and no effect upon the cellular morphology. PNPG was found to be freely water soluble, stable to autoclaving, and to retain biological activity for at least one month in prepared culture media stored under refrigeration.

Aerobiosis↗

Occurrence and pathogenic role of Morganella-Proteus-Providencia group bacteria in human feces.

A total of 2,693 fecal specimens, with 1,422 from healthy persons and 1,271 from patients suffering from enteric diseases, was investigated to isolate species of the Morganella-Proteus-Providencia group and to evaluate the role of these bacteria in intestinal disorders. Most strains were isolated from two media, i.e., blood agar and tryptophan agar. Two of the species were more frequently found in diarrheal cases than in healthy controls. These species were Morganella morganii and Proteus mirabilis. Two new species of Enterobacteriaceae, i.e., Proteus penneri and Providencia rustigianii, were found in 33 and 5 people, respectively. However, these two species were not found more frequently in the diarrheal cases.

Culture Media↗

Methyl mercaptan and dimethyl disulfide production from methionine by Proteus species detected by head-space gas-liquid chromatography.

Head-space gas-liquid chromatography and mass spectrometry were used to detect and identify products formed by Proteus vulgaris, P. mirabilis, P. morganii, and P. rettgeri from a defined medium supplemented with either phenylalanine, methionine, valine, leucine, histidine, lysine, ornithine, threonine, asparagine, aspartic acid, or tryptophan. In a detailed study of the products formed by 68 strains of Proteus spp. from L-methionine, the production of large amounts of both dimethyl disulfide and methyl mercaptan was found to be a characteristic of the genus. Both sulfur products appeared within a few hours of inoculation. Dimethyl disulfide was a more sensitive indicator of growth than the spectrometric determination of optical density. This suggests that it could be useful for the rapid, automated detection of any species of Proteus.

Amino Acids↗

[Sensitivity of Proteus hauseri bacteria to chemotherapeutic preparations depending on the cultivation conditions and on the composition of the nutrient medium].

Sensitivity of 227 Proteus strains isolated from patients was studied comparatively using the agar-diffusion method (disks) and the method of serial dilutions. Marked differences in the numbers of the strains resistant to benzylpenicillin and chloramphenicol were found with the above methods. It was shown that the ingredients of Ploskirev's medium significantly (by 2.8--13.5 times) inhibited the antibacterial activity of streptomycin, neomycin, monomycin, kanamycin, ampicillin and nalidixic acid and had practically no effect on the activity of benzylpenicillin, chloramphenicol and furazolidone. The values of the MIC of the drugs used in the experiment with liquid media correlated with those obtained with Sabouro's medium, which provided recommendation of the latter for determination of Proteus sensitivity by the method of serial dilutions in the solid medium, Cultivation of Proteus at a temperature of 40 degrees C resulted in a decrease of the resistance to most of the drugs tested by (by 3--12.4 times).

Anti-Bacterial Agents↗

[Antibiotic sensitivity of Proteus, Pseudomonas pyocyanea and staphyloccoci isolated from scleroma and ozena patients].

Antibiotic sensitivity of 292 strains of Proteus, 60 strains of Ps, aeruginosa, 309 strains of S. aureus and 88 strains of S. epidermidis isolated from the upper respiratory tract of patients with scleroma and ozena was studied. The cultures of Pr. mirabilis were sensitive to aminoglucosides (54.9-96.2 per cent) and Pr. morganii were sensitive to levomycetin (81.5 per cent) and neomycin (92.6 per cnet). Sensitivity of Pr. vulgaris and Pr. morganii was reliably higher (p less than 0.001) than that of Pr. mirabilis. The strains of Pr. morganii were less sensitive to monomycin (P less than 0.001) and streptomycin (p less than 0.01) as compared to the cultures of other Proteus species tested. The strains of Ps. aeruginosa were sensitive only to gentamicin (90 per cent) and neomycin (81.1 per cent). Most of the strains of S. aureus (85.4-100 per cent) were sensitive to oleadomycin, erythromycin, olemorphocycline, tetraolean, oxacillin, methicillin ceporin, lincomycin, ristomycin, kanamycin, monomycin and gentamicin. Benzylpenicillin (90.8 per cent of the sensitive strains), ampicillin (67.1 per cent), tetracycline (66.7 per cent), levomycetin (68.6 per cent) and streptomycin (38.1 per cent) were less effective. Antibacterial therapy in cases with scleroma and ozena should be directed not only against causative agents of the diseases but also against the microbes developing due to disbacteriosis. Combination of parenteral and local use of the antibiotics in the treatment of chronic clebsiellesis decreased the isolation rate of Proteus and Ps. aeruginosa in the patients.

Anti-Bacterial Agents↗

DNA fingerprinting: the utilization of minisatellite probes to detect a somatic mutation in the proteus syndrome.

Syndromes with localized or segmental abnormalities have been proposed to be the result of a somatic mutation leading to the presence of somatic mosaicism in the tissue. The Proteus syndrome, with its hemihypertrophy, macrodactyly and exostoses, has features which would indicate that the phenotype results from such events. The success of utilizing DNA fingerprint probes to detect somatic mutations in cancer raised the possibility that a similar approach might be successful in an investigation of two patients with the Proteus syndrome. Single band differences were detected with the probe 33.6 in a pair of monozygotic twins discordant for Proteus and in a comparison of tissue from normal and affected areas in another patient. These findings would appear to confirm the hypothesis that the Proteus syndrome results from a somatic mutation. Furthermore, the results indicate that DNA fingerprinting may offer a valuable technique for identifying probes for investigations of similar syndromes.

Abnormalities, Multiple↗

Chaotic behavior in the locomotion of Amoeba proteus.

The locomotion of Amoeba proteus has been investigated by algorithms evaluating correlation dimension and Lyapunov spectrum developed in the field of nonlinear science. It is presumed by these parameters whether the random behavior of the system is stochastic or deterministic. For the analysis of the nonlinear parameters, n-dimensional time-delayed vectors have been reconstructed from a time series of periphery and area of A. proteus images captured with a charge-coupled-device camera, which characterize its random motion. The correlation dimension analyzed has shown the random motion of A. proteus is subjected only to 3-4 macrovariables, though the system is a complex system composed of many degrees of freedom. Furthermore, the analysis of the Lyapunov spectrum has shown its largest exponent takes positive values. These results indicate the random behavior of A. proteus is chaotic and deterministic motion on an attractor with low dimension. It may be important for the elucidation of the cell locomotion to take account of nonlinear interactions among a small number of dynamics such as the sol-gel transformation, the cytoplasmic streaming, and the relating chemical reaction occurring in the cell.

Algorithms↗

Immunocytochemical demonstration of visual pigments in the degenerate retinal and pineal photoreceptors of the blind cave salamander (Proteus anguinus).

Visual pigments in the regressed eye and pineal of the depigmented neotenic urodele, the blind cave salamander (Proteus anguinus anguinus), were studied by immunocytochemistry with anti-opsin antibodies. The study included light- and electron-microscopic investigations of both the eye and the pineal organ. A comparison was made with the black pigmented subspecies Proteus anguinus parkelj (black proteus), which has a normal eye structure. In the retina of the black proteus, we found principal rods, red-sensitive cones and a third photoreceptor type, which might represent a blue- or UV-sensitive cone. Photoreceptors in the regressed eye of the blind cave salamanders from the Planina cave contained degenerate outer segments, consisting of a few whorled discs and irregular clumps of membranes. The great majority of these outer segments showed immunolabelling for the red-sensitive cone opsin and only a few of them were found to be positive for rhodopsin. An even more pronounced degeneration was observed in the photoreceptors of the animals derived from the Otovec doline, which are completely devoid of an outer segment, most of them not even possessing an inner segment. Even in some of these highly degenerate cells, the presence of rhodopsin could be detected in the plasma membrane; however, immunoreactions with antibodies recognizing cone visual pigment were negative. In the pineals of all studied animals, the degenerate photoreceptor outer segments were recognized exclusively by the antibody against the red-sensitive cone opsin. The presence of immunopositive visual pigments indicates the possibility of a retained light sensitivity in the blind cave salamander photoreceptors.

Animals↗

Studies of the DNA of Amoeba proteus and Amoeba indica.

1. The G + C content of the DNAs of Amoeba proteus and Amoeba indica was determined by buoyant density gradient centrifugation. 2. Generally, different clones of the same strains did not differ in the G + C content of their main DNA or their satellite DNA. 3. The G + C content of the main DNA of the TD and XD strains of A. proteus were the same (25.3 and 29.3% respectively) as were the G + C contents of the satellite DNA (54.7 and 61.8% respectively). 4. The main and satellite DNAs are reversed for the F strain of A. proteus and the I strain of A. indica; the G + C contents of the main DNA being 57.4 and 59.8% respectively and those of the satellite DNA being 23.5 and 28.4% respectively. 5. The heterologous hybrids produced by nuclear transplantation between the A strain of A. proteus and the I strain of A. indica have the same G + C content in their main and satellite DNAs as do their cytoplasmic parents. 6. The data suggest that the occurrence of phenotypic variations does not necessarily mean an alteration in the G + C content of either main or satellite DNA. 7. There is also evidence that the G + C content of these DNAs in under cytoplasmic control.

Amoeba↗

Mechanics and control of the cytoskeleton in Amoeba proteus.

Many models of the cytoskeletal motility of Amoeba proteus can be formulated in terms of the theory of reactive interpenetrating flow (Dembo and Harlow, 1986). We have devised numerical methodology for testing such models against the phenomenon of steady axisymmetric fountain flow. The simplest workable scheme revealed by such tests (the minimal model) is the main preoccupation of this study. All parameters of the minimal model are determined from available data. Using these parameters the model quantitatively accounts for the self assembly of the cytoskeleton of A. proteus: for the formation and detailed morphology of the endoplasmic channel, the ectoplasmic tube, the uropod, the plasma gel sheet, and the hyaline cap. The model accounts for the kinematics of the cytoskeleton: the detailed velocity field of the forward flow of the endoplasm, the contraction of the ectoplasmic tube, and the inversion of the flow in the fountain zone. The model also gives a satisfactory account of measurements of pressure gradients, measurements of heat dissipation, and measurements of the output of useful work by amoeba. Finally, the model suggests a very promising (but still hypothetical) continuum formulation of the free boundary problem of amoeboid motion. by balancing normal forces on the plasma membrane as closely as possible, the minimal model is able to predict the turgor pressure and surface tension of A. proteus. Several dynamical factors are crucial to the success of the minimal model and are likely to be general features of cytoskeletal mechanics and control in amoeboid cells. These are: a constitutive law for the viscosity of the contractile network that includes an automatic process of gelation as the network density gets large; a very vigorous cycle of network polymerization and depolymerization (in the case of A. proteus, the time constant for this reaction is approximately 12 s); control of network contractility by a diffusible factor (probably calcium ion); and control of the adhesive interaction between the cytoskeleton and the inner surface of the plasma membrane.

Amoeba↗

Rapid detection of Obesumbacterium proteus from yeast and wort using polymerase chain reaction.

AIMS: To design and evaluate PCR primers for the rapid detection of Obesumbacterium proteus. METHODS AND RESULTS: The 16S rDNA from a wild-type Obesumbacterium proteus biotype II isolate was sequenced and the resultant data used to produce specific primers for this organism. These primers discriminated between biotype I (nonbrewery) and biotype II isolates. In addition, the primers were able to detect Obesumbacterium proteus in wort and in yeast slurries in the presence of competitive bacteria. The primers were tested against a range of other beer spoilage bacteria for any cross-reactions. None were detected. CONCLUSIONS: Obesumbacterium proteus primers can detect this contaminant without generating cross-reactions to related species. SIGNIFICANCE AND IMPACT OF THE STUDY: The primers generated in this study can now be used for PCR detection assays that will contribute to early detection of this important process contaminant.

Beer↗

Defense function of pigment granules in the ciliate Blepharisma japonicum against two predatory protists, Amoeba proteus (Rhizopodea) and Climacostomum virens (Ciliata).

The defense function of pigment granules in the red ciliate Blepharisma japonicum against two predatory protists, Amoeba proteus and Climacostomum virens, was investigated by (1) comparing normally-pigmented and albino mutant cells of B. japonicum as the prey of these predators and (2) comparing resistance of the predators to blepharismin, the toxic pigment contained in the pigment granules of B. japonicum. Normally pigmented cells which contained more blepharismin than albino cells were less vulnerable to A. proteus than albino cells, but not to C. virens. C. virens was more resistant than A. proteus to the lethal effect of blepharismin. The results indicate that pigment granules of B. japonicum function as defense organelles against A. proteus but not against C. virens and suggest that successful defense against a predator depends on the susceptibility of the predator to blepharismin.

Amoeba↗

Ocular development and involution in the European cave salamander, Proteus anguinus laurenti.

The anatomy and development of the eye of Proteus anguinus are described. The relationships between organogenesis of the eye in embryos and larva and its involution in the young and the adult are discussed. The availability (in breeding cultures) of a significant number of Proteus embryos (which are normally rare) allowed experimental analysis of the effects of light, xenoplastic differentiation and thyroid hormones on the development of the eye. The results of this study suggest that development and involution of the eye of Proteus are controlled by genetic factors which are not greatly influenced by environment, and one can, therefore, consider the microphthalmy of Proteus as a relict characteristic which is the result of a specific development with disturbance of the normal ontogenic process.

Age Factors↗

Role of internal fixation in infection of open fractures: studies with Staphylococcus aureus and Proteus mirabilis.

Studies of infections in open fractures are described. The hamster was chosen as the experimental model. Osteotomies of the femur were created with an osteotomy saw. Some osteotomies were left to heal and others were fixed with an 0.9 mm K-wire. The infection rates in fixed and unfixed fractures were compared. The first group of hamsters with fixed and unfixed osteotomies was returned to cages with open wounds. There was no difference in the infection rate at 2 weeks. The second group was deliberately contaminated with Staphylococcus aureus and then returned to cages with open wounds. In this group, the infection rate at 2 weeks was lower in the internally-fixed fractures than in the unfixed fractures. The third group was deliberately contaminated with the gram negative organism Proteus mirabilis. In these animals, the infection rate was increased in the presence of the internal fixation device. The fourth group was deliberately contaminated with both Staphylococcus aureus and Proteus mirabilis. The infection rate in these animals was very high; Proteus was recovered from those animals with internal fixation and Staphylococcus was recovered from those animals without internal fixation. These studies in the hamster document the usefulness of this animal as an inexpensive and reproducible model for studying infection of open fractures. The hamsters tolerated the procedure well, and wound and fracture healing progressed satisfactorily.

Animals↗

Correlation between anti-Proteus antibodies and isolation rates of P. mirabilis in rheumatoid arthritis.

In a survey of 89 RA patients, carried out under code, Proteus mirabilis was isolated from the urine of 63% (47/75) of female (P < 0.001) and 50% (7/14) of male patients (P < 0.001), compared to a frequency of isolation in healthy women of 32% (38/119) and 11% (13/115) in healthy men. There was no significant difference in isolation rates between 37 non-RA patients and healthy controls. Sera from 20 patients with RA and 20 healthy controls were tested against P. mirabilis and Escherichia coli by an enzyme-linked immunosorbent assay. Antibodies against P. mirabilis but not to E. coli were significantly higher in the RA patients than in healthy controls (P < 0.001). Furthermore, a positive correlation was found between high anti-Proteus antibody levels in serum samples and the number of Proteus colony-forming units obtained from urine specimens of the 20 RA patients (r = +0.714, P < 0.001). These results support the suggestion of an aetiopathogenic role for P. mirabilis in RA.

Adult↗