is a bacterium belonging to the class
. It causes acute and chronic infections of the urogenital tract. The main features of this bacterium are an absence of cell wall and a reduced genome size ...(517-622 protein-encoding genes). Previously, we have isolated morphologically unknown
colonies called micro-colonies (MCs) from the serum of patients with inflammatory urogenital tract infection.
MCs are functionally different from the typical colonies (TCs) in terms of metabolism and cell division.
To determine the physiological differences between MCs and TCs of
and elucidate the pathways of formation and growth of MCs by a comparative proteomic analysis of these two morphological forms.
LC-MS proteomic analysis of TCs and MCs using an Ultimate 3000 RSLC nanoHPLC system connected to a QExactive Plus mass spectrometer.
The study of the proteomic profiles of
colonies allowed us to reconstruct their energy metabolism pathways. In addition to the already known pentose phosphate and arginine deamination pathways,
can utilise ribose phosphate and deoxyribose phosphate formed by nucleoside catabolism as energy sources. Comparative proteomic HPLC-MS analysis revealed that the proteomic profiles of TCs and MCs were different. We assume that MC cells preferably utilised deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Utilisation of deoxyribonucleosides is less efficient as compared with that of ribonucleosides and arginine in terms of energy production. Thymidine phosphorylase DeoA is one of the key enzymes of deoxyribonucleosides utilisation. We obtained a DeoA overexpressing mutant that exhibited a phenotype similar to that of MCs, which confirmed our hypothesis.
In addition to the two known pathways for energy production (arginine deamination and the pentose phosphate pathway)
can use deoxyribonucleosides and ribonucleosides. MC cells demonstrate a reorganisation of energy metabolism: unlike TC cells, they preferably utilise deoxyribonucleosides, particularly thymidine, as an energy source rather than arginine or ribonucleosides. Thus MC cells enter a state of energy starvation, which helps them to survive under stress, and in particular, to be resistant to antibiotics.
Improving the management of children with asthma associated with mycoplasma infection is important. Aim: To study the duration of the persistence of antigens, and DNA in a free state, in the ...structures of circulating immune complexes (CICs) and living cells of
and
in children with asthma. In total, 205 children with asthma from 1 to 14 years were observed. The reaction of aggregate-hemagglutination (AHAA), the direct immunofluorescence reaction (DIF), the reaction of the polymerase chain reaction (PCR), and the culture method were used. In addition, 47 children were re-examined 1.5 months after the treatment of mycoplasma infection with azithromycin. The number of samples positive for antigens and DNA in the free state and in the structures of CICs significantly decreased. Then, 50 blood serum samples containing
antigens, and 50 samples containing
antigens were analyzed by culture method.
was isolated in 21 (65.5%) of 32 samples containing DNA.
was isolated from antigen-positive samples in nine cases. The presented data indicate the long-term persistence of antigens, and DNA of mycoplasma cells in the free state, in the structure of CICs, as well as in the form of "microcolonies". A high level of CICs can be used to predict the course of the disease and the response to therapy.
The
class unites cell wall lacking bacteria many of which are membrane parasites and opportunistic bacteria.
. This study describes a novel morphological form found in the five species belonging to ...the bacterial class Mollicutes, and referred to as microcolonies (MCs).
MCs were obtained as described below and characterized with bacteriological and immunological methods, and microscopy.
In contrast to typical colonies (TCs), MCs are characterized by tiny propeller-shaped colonies formed by rod-like cells tightly packed in parallel rows. These colonies were observed within routinely cultivated cultures of type strains 7-12 days post-plating. Rod-like cells were visualized using a scanning electron microscope within TCs with a 'fried-egg-like' appearance. MCs were not observed to revert to TCs. MCs were resistant to antibiotics and other treatments effective against TCs. Pure MC cultures were generated
by treatment of
cultures with hyperimmune serum, antibiotics or argon non-thermal plasma. MCs of
strain H-34 were characterized in detail to confirm that they belonged to that species. MCs tested positive via PCR with
-specific primers, direct fluorescence and epifluorescence tests, and Western blotting with the camel-derived nanobody aMh-FcG2a, which is specific to the MH3620 transporter protein. Meanwhile, MCs behaved differently in standard bacteriological tests. Pure MC cultures were also isolated directly from clinical samples of the serum, synovial liquid and urine of patients within flammatory urogenital tract diseases, asthma or arthritis. In total, 79 independent MC cultures were isolated from clinical samples including
(
=70),
(
=2),
(
=2) and
spp. (
=5).
. MCs play an unknown role in infection pathology and display prominent antibiotic resistance, making them a challenge for the future studies on Mollicutes.
Mycoplasma hominis is an opportunistic bacterium that can cause acute and chronic infections of the urogenital tract. This bacterium, like all other Mycoplasma species, is characterized by the ...reduced genome size, and, consequently, reduction of the main metabolic pathways. M. hominis cells cannot effectively use glucose as a carbon and energy source. Therefore, the main pathway of energy metabolism is the arginine dihydrolase pathway. However, several bacteria can use nucleosides as the sole energy source. Biochemical studies using Salmonella typhimurium have shown that three enzymes (thymidine phosphorylase, phosphopentose mutase and deoxyribose-phosphate aldolase) are involved in the thymidine catabolic pathway. All these enzymes are present in M. hominis. For understanding changes in the energy metabolism of M. hominis we performed shotgun proteome analysis of M. hominis cells in liquid medium with arginine or thymidine as a carbon source. LC-MS analysis was performed with an Ultimate 3000 Nano LC System (Thermo Fisher Scientific) coupled to a Q Exactive HF benchtop Orbitrap mass spectrometer (Thermo Fisher Scientific) via a nanoelectrospray source (Thermo Fisher Scientific). Data are available via ProteomeXchange with identifier PXD018714 (https://www.ebi.ac.uk/pride/archive/projects/PXD018714).
The origin of chronic inflammation preceding the development of prostate cancer (PCa) remains unknown. We investigated possible involvement of mycoplasma infection in PCa by screening prostate ...biopsies from two groups of Russian men undergoing PCa diagnosis. M. hominis was detected by standard PCR in 15% of the 125 patients in the first group and by quantitative real-time PCR in 37.4% of the 123 men in the second group. In both groups, stratification of patients according to diagnosis showed that M. hominis was present at three times higher frequency in patients with PCa than in those with benign prostatic hyperplasia. No M. hominis was detected in the prostates of 27 men without detectable prostate disease. In addition, PCa-positive men had higher titers of antibodies against M. hominis and average PSA levels were higher in M. hominis-positive men. These data, together with previous observations linking mycoplasma infection with cell transformation, genomic instability and resistance to apoptosis, suggest that M. hominis infection may be involved in PCa development and may, therefore, be a potential PCa marker and/or target for improved prevention and treatment of this disease.
This study evaluated the safety and efficacy of recombinant human intestinal trefoil factor (rhITF) administered as topical oral spray for prevention and treatment of chemotherapy-induced oral ...mucositis (OM).
Ninety-nine patients with colorectal cancer who had moderate to severe OM (WHO grade >or= 2) in the first cycle of chemotherapy were randomly assigned to receive either placebo, rhITF 10 mg/mL (ie, low dose), or rhITF 80 mg/mL (ie, high dose) by oral spray (300 microL, eight times each day) for 14 consecutive days in the second chemotherapy cycle. Patients were assessed on days 1, 3, 5, 7, 10, 12, 14, and 21 (+/- 2 days for the last assessment) for safety and for OM incidence and severity.
Treatment of patients at high risk for developing OM with low- or high-dose rhITF significantly reduced the amount of incidence (75% to 81%; low-dose rhITF P < .001; high-dose rhITF P = .002). Frequencies of WHO grade >or= 2 OM in the placebo, low-dose rhITF, and high-dose rhITF groups were 48.5%, 9.1%, and 12.1%, respectively. Assessment of the area under the curve revealed statistically significant reductions in OM severity in the rhITF-treated groups versus placebo. Only a minority of patients (6.1%) reported treatment-emergent adverse events (TEAEs), all of which were mild to moderate in intensity and resolved without sequelae. The incidence of TEAEs was not significantly different among treatment groups.
rhITF oral spray formulation was safe and effective when used for the reduction of chemotherapy-associated OM in patients with colorectal cancer. Patients exhibited high compliance in dosing administration. Future clinical study is planned to develop this drug for use in OM management in patients with cancer.
Interactions of Acholeplasma laidlawii cells with mouse spleen lymphocytes Tarshis, M.A. (Moscow State Univ. (USSR). Dept. of Biochemistry); Ladygina, V.G; Migoushina, V.L ...
Zentralblatt für Bakteriologie, Mikrobiologie und Hygiene. 1. Abt. Originale, A, Medizinische Mikrobiologie, Infektionskrankheiten und Parasitologie,
01/1981, Letnik:
250, Številka:
1-2
Journal Article
Interaction between Acholeplasma laidlawii cells labelled with oleic acid and mouse spleen lymphocytes depended on the time of incubation, on the temperature and on the quantitative ratio between ...both cells. Uncouplers and EDTA did not influence the intensity of attachment. The resistance of binding to cytochalasin B amd glutaraldehyde as well as localization of A. laidlawii antigens on the lymphocyte surface and experiments with 14C uridine-labelled mycoplasmas are an evidence against the participation of pinocytosis in this interaction. Prolonged attachment of intact A. laidlawii to washed lymphocytes can be excluded on the basis of an extremely low amount of CFU recovered from disrupted lymphocytes as well as by experiments with uridine-labelled A laidlawii. Specific receptors didn't take part in the binding, because proteolytic enzymes and neuraminidase treatment proved to be nonefficient. Increased binding of lymphocytes with liposomes prepared from mycoplasma lipids as well as the transfer of cholesterol from lymphocyte membrane to mycoplasma membrane demonstrate the participation of membrane lipids in this binding. It should also be mentioned that after the attachment between both cell types and fusion of A. laidlawii cells with lymphocytes takes place. The transfer of unsaturated fatty acids from mycoplasmas into lymphocyte membrane as well as lymphocyte membrane cholesterol into mycoplasma membranes are the consequence of fusion between both cells. The experiments with uncharged hydrophobic fluorescent probe 4-DMC are the direct proof of fusion and mutual exchange of lipid membrane components.
Transport of two non-metabolized carbohydrates (3-O-methyl-D-glucose, 3-O-MG, and 2-deoxy-D-glucose, 2-DG) into mouse spleen lymphocytes after their interaction with Acholeplasma laidlawii cells has ...been studied. Incubation of A. laidlawii cells and particularly the liposomes prepared from A. laidlawii membrane lipids enhances the rates of the both carbohydrates transport. This treatment resulted in increasing of the Vmax values of 3-O-MG and 2-DG without changing the Km values. This stimulation can be explained by the increasing of the mobility of membrane carbohydrates carriers as a result of the exchange of lipid components between Acholeplasma and lymphocyte membranes. Actually, it has been shown that liposomes derived from A. laidlawii cells grown on the medium with great amount of unsaturated oleic acid stimulate the transport activity more actively than liposomes prepared from the cells grown on the medium with bovine serum or with oleic acid plus cholesterol. It should be suggested that an activation of carbohydrates transport into lymphocytes caused by alteration of the carriers lipid microenvironment.
The duration of Acholeplasma laidlawii antigen persistence in mice, resistant to Rausher leukemia virus, after infection with both A. laidlawii cells and Rausher virus has been studied. The antigen ...persistence was accompanied by marked depression of immune response which was especially severe in case of mixed acholeplasmavirus infection. Such immunosuppression and observed infiltration of the spleen with immature leukemic cells can be regarded as a preleukosis. Immunosuppression was accompanied by an increase of the transport of carbohydrates inthe lymphocytes. This stimulation an be explained by the exchange of lipid components between acholeplasma and lymphocyte membranes resulted in increase of lymphocyte membrane fluidity, or it may be due to the mitogenic effect of A. laidlawii cells and virus, accompanied by the same membrane effect.
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