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[Adoptive immunotherapy for pulmonary tuberculosis caused by multi-resistant bacteria using autologous peripheral blood leucocytes sensitized with killed Mycobacterium tuberculosis bacteria].

A patient with pulmonary tuberculosis caused by bacteria resistant to various anti-microbial agents was treated with adoptively transferred autologous peripheral blood leucocytes (PBL) sensitized with killed Mycobacterium tuberculosis organisms in vitro. The 32-year-old man was admitted to our hospital from National Sanitarium Okinawa Hospital with weight loss, high fever, and rapid aggravation on chest X-ray. Patient's PBL obtained by leukapheresis and separated with Ficoll-Hypaque solution were cultured with killed Mycobacterium tuberculosis bacteria of 0.4 microgram per ml at 1 x 10(6) cells per ml for 7 days in media containing 0.5 U recombinant 1L-2 per ml. After incubation, PBL were layered and centrifuged on Ficoll-Hypaque solution and washed three times with saline. PBL (1-3 x 10(8)) were combined and concentrated for infusion in 20 to 30 ml saline. After injection, patient displayed fever and transitory drop of PaO2. Although the patient did not have an improved on chest X-ray, his fever was alleviated, weight was increased, accelerated ESR was slightly improved, and the number of organisms in sputum (Number of Gaffky) temporarily decreased. Adoptive immunotherapy using the autologous PBL which were sensitized with killed bacteria may be an effective anti-tuberculous immunotherapy.

Adult↗

The role of easily released substances (ERS) from bacteria in physiologic solution in natural immunity. VI. Induction of opsonins for different genera of bacteria by ERS from Staph. aureus oxford.

ERS obtained from Staph. aureus by mild extraction with physiologic saline solution possess activity of inducing nonspecific opsonins which react with homologous bacteria and with related Gram-negative bacteria. ERS from Staph. aureus weakly stimulated precipitins and antibodies active in passive hemagglutination and induced a marked rise in agglutinin titers especially for the homologous strain and Gram-negative S. typhimurium bacilli. N-acetyl-D-glucosamine absorbed the opsonizing factor for various bacteria with heterologous activity.

Acetylglucosamine↗

Isolation and characterization of gram-positive cyanophycin-degrading bacteria-kinetic studies on cyanophycin depolymerase activity in aerobic bacteria.

This study is the first report on the extracellular degradation of cyanophycin (CGP) by Gram-positive bacteria. Three different Gram-positive bacteria were isolated from forest soil that were able to utilize CGP as the sole carbon source for growth. The isolates were assigned to species of the genera Bacillus and Micromonospora. From one of the isolates, which was taxonomically affiliated as Bacillus megaterium strain BAC19, the extracellular CGP depolymerase (extracellular CGPase; CphEBm) was purified to electrophoretic homogeneity by fast protein liquid chromatography and affinity binding to an arginine-agarose column. The purified enzyme was specific for hydrolytic cleavage of CGP, and inhibitor studies indicated that CphEBm is a serine-type peptidase. As CGP degradation products, (beta-Asp-Arg)2 tetrapeptides in addition to beta-Asp-Arg dipeptides occurred, which were identified by electrospray ionization mass spectrometry analysis. Furthermore, a novel quantitative enzyme assay was developed for kinetic studies on CGP depolymerases. For CphEBm, as well as for the extracellular CGPase of Pseudomonas anguilliseptica strain BI (CphEPa), KM values of 2.2 and 1.0 microM, respectively, for CGP were determined.

Bacillus↗

Comparative surface-to-hand and fingertip-to-mouth transfer efficiency of gram-positive bacteria, gram-negative bacteria, and phage.

AIMS: To determine the transfer efficiency of micro-organisms from fomites to hands and the subsequent transfer from the fingertip to the lip. METHODS AND RESULTS: Volunteers hands were sampled after the normal usage of fomites seeded with a pooled culture of a Gram-positive bacterium (Micrococcus luteus), a Gram-negative bacterium (Serratia rubidea) and phage PRD-1 (Period A). Activities included wringing out a dishcloth/sponge, turning on/off a kitchen faucet, cutting up a carrot, making hamburger patties, holding a phone receiver, and removing laundry from the washing machine. Transfer efficiencies were 38.47% to 65.80% and 27.59% to 40.03% for the phone receiver and faucet, respectively. Transfer efficiencies from porous fomites were <0.01%. In most cases, M.luteus was transferred most efficiently, followed by phage PRD-1 and S. rubidea. When the volunteers' fingertips were inoculated with the pooled organisms and held to the lip area (Period B), transfer rates of 40.99%, 33.97%, and 33.90% occurred with M. luteus, S. rubidea, and PRD-1, respectively. CONCLUSIONS: The highest bacteral transfer rates from fomites to the hands were seen with the hard, non-porous surfaces. Even with low transfer rates, the numbers of bacteria transferred to the hands were still high (up to 10(6) cells). Transfer of bacteria from the fingertip to the lip is similar to that observed from hard surfaces to hands. SIGNIFICANCE AND IMPACT OF THE STUDY: Infectious doses of pathogens may be transferred to the mouth after handling an everyday contaminated household object.

Adolescent↗

Reverse effect of gram-positive bacteria vs. gram-negative bacteria on adjuvant-induced arthritis in germfree rats.

Germfree (GF) F344 rats developed severe adjuvant-induced arthritis with a 100% incidence after a single intradermal injection of heat-killed Mycobacterium bovis (BCG). Specific pathogene-free (SPF) rats developed less severe arthritis with a lower incidence. The rats colonized with Escherichia coli or Bacteroides developed mild disease comparable to that in SPF rats. The rats colonized with Bifidobacterium, Propionibacterium acnes, Lactobacillus casei, L. fermentum, L. murini, and L. acidophilus developed more severe disease than that in GF rats. Furthermore, the rats colonized with a mixture of E. coli and the above lactobacilli developed very mild disease similar to that in SPF rats. These results suggest that gram-negative bacteria, such as E. coli and Bacteroides, may suppress the disease, possibly through their lipopolysaccharides, and may be responsible for the lower susceptibility of SPF rats; gram-positive bacteria, such as Bifidobacterium, P. acnes, and lactobacilli, may enhance the disease, possibly through their peptidoglycans; and E. coli may play a dominant role in modulating the development of adjuvant-induced arthritis.

Animals↗

Gram-negative bacteria produce membrane vesicles which are capable of killing other bacteria.

Naturally produced membrane vesicles (MVs), isolated from 15 strains of gram-negative bacteria (Citrobacter, Enterobacter, Escherichia, Klebsiella, Morganella, Proteus, Salmonella, and Shigella strains), lysed many gram-positive (including Mycobacterium) and gram-negative cultures. Peptidoglycan zymograms suggested that MVs contained peptidoglycan hydrolases, and electron microscopy revealed that the murein sacculi were digested, confirming a previous modus operandi (J. L. Kadurugamuwa and T. J. Beveridge, J. Bacteriol. 174:2767-2774, 1996). MV-sensitive bacteria possessed A1alpha, A4alpha, A1gamma, A2alpha, and A4gamma peptidoglycan chemotypes, whereas A3alpha, A3beta, A3gamma, A4beta, B1alpha, and B1beta chemotypes were not affected. Pseudomonas aeruginosa PAO1 vesicles possessed the most lytic activity.

Antibiosis↗

Metal accumulation by bacteria with particular reference to dissimilatory sulphate-reducing bacteria.

Dissimilatory sulphate-reducing bacteria, genera Desulfovibrio and Desulfotomaculum, exhibit a superior ability, over assimilatory organisms, to extract three amounts of metals from culture media. This property does not appear to be solely a function of the presence of H2S. In media containing elevated amounts of Fe, electron dense particles, provisionally identified as FeS, are deposited within the cells of dissimilatory bacteria.

Bacillaceae↗

Post-genomics of lactic acid bacteria and other food-grade bacteria to discover gut functionality.

Recent years have seen an explosion in the number of complete or almost complete genomic sequences of lactic acid bacteria and other food-grade bacteria that are used in functional foods to increase the health of the consumer. These have been instrumental in the development of functional, comparative and other post-genomics approaches that provide the possibility to detect, unravel and understand their functionality in the human intestinal tract. In conjunction with other high-throughput approaches, these advances can be exploited in the functional food innovation cycle for developing new or designed probiotic and other bacterial products that impact gut health.

Animals↗

The use of a solid adsorber resin for enrichment of bacteria with toxic substrates and to identify metabolites: degradation of naphthalene, O-, and m-xylene by sulfate-reducing bacteria.

Anaerobic sulfate-reducing bacteria were enriched from contaminated aquifer samples with naphthalene, o-, and m-xylene as sole carbon and energy source in the presence of Amberlite-XAD7, a solid adsorber resin. XAD7 served as a substrate reservoir maintaining a constantly low substrate concentration in the culture medium. In equilibration experiments with XAD7, the aromatic hydrocarbons needed up to 5 days to achieve equilibrium between the water and the XAD7 phase. The equilibrium concentration was directly correlated with the amount of added substrate and XAD7. In the enrichments presented here, XAD7 and aromatic hydrocarbons were adjusted to maintain substrate concentrations of 100 microM m-, or o-xylene, or 50 microM naphthalene. After five subsequent transfers, the three cultures were able to grow with higher substrate concentrations in the absence of XAD7 although they grew best with lower hydrocarbon concentrations. Two new xylene-degrading cultures were obtained that could not utilise toluene as carbon source. O-xylene was degraded anaerobically by a culture, which could also oxidise m-xylene but not p-xylene. Eighty-three percent of the electrons from o-xylene oxidation were recovered in the produced sulfide, indicating a complete oxidation to CO2. Another sulfate-reducing enrichment culture oxidised m-xylene completely to CO2 but not o-, or p-xylene. A naphthalene-degrading sulfate-reducing enrichment culture oxidised naphthalene completely to CO2. Metabolites of naphthalene degradation were recovered from the XAD7 phase and subjected to GC/MS analysis. Besides the metabolites 2-naphthoic acid and decahydro-2-naphthoic acid which were identified by the mass spectrum and coelution with chemically synthesised reference compounds, the reduced 2-naphthoic acid derivatives 5,6,7,8-tetrahydro-2-naphthoic acid and octahydro-2-naphthoic acid were tentatively identified by their mass spectra. Cultivation of bacterial cultures in the presence of XAD7 and subsequent derivatisation and extraction of metabolites directly from the solid XAD7 resin provides a new method for the isolation of sensitive bacteria and identification of metabolites.

Acrylic Resins↗

Nitrite reductase activity of sulphate-reducing bacteria prevents their inhibition by nitrate-reducing, sulphide-oxidizing bacteria.

Sulphate-reducing bacteria (SRB) can be inhibited by nitrate-reducing, sulphide-oxidizing bacteria (NR-SOB), despite the fact that these two groups are interdependent in many anaerobic environments. Practical applications of this inhibition include the reduction of sulphide concentrations in oil fields by nitrate injection. The NR-SOB Thiomicrospira sp. strain CVO was found to oxidize up to 15 mM sulphide, considerably more than three other NR-SOB strains that were tested. Sulphide oxidation increased the environmental redox potential (Eh) from -400 to +100 mV and gave 0.6 nitrite per nitrate reduced. Within the genus Desulfovibrio, strains Lac3 and Lac6 were inhibited by strain CVO and nitrate for the duration of the experiment, whereas inhibition of strains Lac15 and D. vulgaris Hildenborough was transient. The latter had very high nitrite reductase (Nrf) activity. Southern blotting with D. vulgaris nrf genes as a probe indicated the absence of homologous nrf genes from strains Lac3 and Lac6 and their presence in strain Lac15. With respect to SRB from other genera, inhibition of the known nitrite reducer Desulfobulbus propionicus by strain CVO and nitrate was transient, whereas inhibition of Desulfobacterium autotrophicum and Desulfobacter postgatei was long-lasting. The results indicate that inhibition of SRB by NR-SOB is caused by nitrite production. Nrf-containing SRB can overcome this inhibition by further reducing nitrite to ammonia, preventing a stalling of the favourable metabolic interactions between these two bacterial groups. Nrf, which is widely distributed in SRB, can thus be regarded as a resistance factor that prevents the inhibition of dissimilatory sulphate reduction by nitrite.

Antibiosis↗

Novel bacteria degrading N-acylhomoserine lactones and their use as quenchers of quorum-sensing-regulated functions of plant-pathogenic bacteria.

Bacteria degrading the quorum-sensing (QS) signal molecule N-hexanoylhomoserine lactone were isolated from a tobacco rhizosphere. Twenty-five isolates degrading this homoserine lactone fell into six groups according to their genomic REP-PCR and rrs PCR-RFLP profiles. Representative strains from each group were identified as members of the genera Pseudomonas, Comamonas, Variovorax and Rhodococcus: all these isolates degraded N-acylhomoserine lactones other than the hexanoic acid derivative, albeit with different specificity and kinetics. One of these isolates, Rhodococcus erythropolis strain W2, was used to quench QS-regulated functions of other microbes. In vitro, W2 strongly interfered with violacein production by Chromobacterium violaceum, and transfer of pathogenicity in Agrobacterium tumefaciens. In planta, R. erythropolis W2 markedly reduced the pathogenicity of Pectobacterium carotovorum subsp. carotovorum in potato tubers. These series of results reveal the diversity of the QS-interfering bacteria in the rhizosphere and demonstrate the validity of targeting QS signal molecules to control pathogens with natural bacterial isolates.

4-Butyrolactone↗

Expansion of the host range of coliphage P1 and gene transfer from enteric bacteria to other gram-negative bacteria.

The bacterial host range of coliphage P1 was extented by using the heat-inducible phage P1clr100KM. A gene for kanamycin resistance was transferred from Escherichia coli to members of the family Enterobacteriaceae and some other genera of gram-negative bacteria. P1 phage was produced by thermal induction from the lysogens of all these kanamycin-resistant bacteria except some strains.

Bacteria↗

Aerobic heterotrophic bacteria indigenous to pH 2.8 acid mine water: predominant slime-producing bacteria in acid streamers.

Five gram-negative bacteria, two gram-positive bacteria, and one yeast were isolated from "acid streamers" taken from acid mine water. One gram-positive rod which has been tentatively identified as a Bacillus species appeared to be the predominant organism in the streamers. This isolate produced copious amounts of extracellular polymer at 10 C in the laboratory and was considered to be the primary source of polymer in the "acid streamer" slime matrix. The organism grew slowly at pH 2.8 in mine water media, but the optimal pH was approximately 7.0.

Bacillus↗

[Bacteria isolated from surgical infections and their susceptibilities to antimicrobial agents. Special references to bacteria isolated between April 1998 and March 1999].

The annual multicenter studies on isolated bacteria from infections in general surgery and their antimicrobial susceptibility have been conducted in Japan since July 1982. This paper describes the results obtained in fiscal 1998 (from April 1998 to March 1999). The number of cases investigated as objectives was 225 for one year. A total of 429 strains (121 strains from primary infections and 308 strains from postoperative infections) were isolated from 183 cases (81.3% of total cases). In primary infections, the isolation rates of anaerobes and Escherichia coli were higher than in postoperative infections, while in postoperative infections, those of Gram-positive aerobes and Pseudomonas aeruginosa were higher than in primary infections. On the whole, among Gram-positive aerobes, the isolation rate of Enterococcus faecalis was the highest, followed by Staphylococcus aureus with high frequency in isolation from postoperative infections. Among Gram-positive anaerobes, Peptostreptococcus spp. and Streptococcus spp. were predominantly isolated. Among Gram-negative aerobes, E. coli, P. aeruginosa, Klebsiella pneumoniae and Enterobacter cloacae were frequently isolated. Among Gram-negative anaerobes, Bacteroides fragilis group was the majority of isolates. In primary infections, the percentage of Gram-negative aerobes has gradually increased since fiscal 1995 or 1996 with these years as the turning point, while those of Gram-positive and Gram-negative anaerobes have gradually declined. In postoperative infections, the percentage of Gram-negative anaerobes has increased continuously since the mid-1980s. The percentage of MRSA among S. aureus rose to 89.7%, which was the highest level since the beginning of this study. The susceptibilities of B. fragilis, which did not show apparent changes, were recognized to have decreased against cephems in fiscal 1998. Among other bacteria in B. fragilis group, development of resistance to cephems has continued on a long-term basis since the mid-1980s. E. coli and K. pneuminiae have obviously not changed in susceptibilities, however, the susceptibilities of isolated strains in fiscal 1998 against high-generation cephems, oxacephems and monobactams have declined. We found neither vancomycin-resistant nor teicoplanin-resistant strains of S. aureus and Enterococcus spp.

Anti-Bacterial Agents↗

[Bacteria isolated from surgical infections and their susceptibilities to antimicrobial agents. Special references to bacteria isolated between April 1999 and March 2000].

The annual multicenter studies on isolated bacteria from infections in general surgery and their antimicrobial susceptibility have been conducted in Japan since July 1982. In this paper, the results obtained in the academic year 1999 (from April 1999 to March 2000) have been summarized. Two hundred seven cases were investigated, and 411 strains were isolated from 169 cases (81.6%). Of those strains, 184 and 227 strains were from primary infections and postoperative infections, respectively. In primary infections, the isolation rates of anaerobes, Streptococcus spp., and Escherichia coli were higher than in postoperative infections, while in postoperative infections, those of Gram-positive aerobes were higher than in primary infections. Staphylococcus aureus were most frequently isolated among Gram-positive aerobes, Peptostreptococcus prevotii among Gram-positive anaerobes, E. coli among Gram-negative aerobes, and Bacteroides fragilis among Gram-negative anaerobes. In primary infections, the percentage of Gram-negative aerobes, which gradually increased by the year 1998, decreased in the year 1999. The percentage of Gram-negative anaerobes increased, while that of Gram-negative bacteria was equivalent to that in the last year. In postoperative infections, the percentage of Gram-negative anaerobes, which continuously increased after the year 1990, decreased, while that of Gram-positive aerobes, which decreased in the last year, increased. Methicillin-resistant S. aureus accounted for 70.7% of S. aureus (41 strains). Either the number of strain or the percentage of MRSA decreased. The susceptibilities of E. coli and Klebsiella pneumoniae decreased against third and forth generation cephems, oxacephems, and monobactams. The susceptibilities of P. aeruginosa to carbapenems tend to decrease after the year 1997. S. aureus showed good susceptibilities to the tested drugs including arbekacin, vancomycin, and teicoplanin.

Anti-Bacterial Agents↗

[Problems in the use of radioactively labelled bacteria in experiments. 2. Quantitative method for the evaluation of pathogens in the calf lung by means of labelled bacteria after aerosol or intratracheal administration].

By means of two aerosol apparatures radioactive labelled Pasteurella-multocida-germs were given to calves via a respiratory mask. Labelling of Pasteurellae was done internal by using special nutritive media with Fe-59 or P-32. In a period of 10-25 minutes 5 X 10(9)-10(10) labelled bacteria were given to the animals for inhalation. Another animal group was injected intratrachealy with the same quantity of labelled germs. Immediately after termination of germ application the animals were killed and dissected. After solution or homogenization, respectively of the total lung tissue radioactivity was determined by liquid-scintillation-counting (Fe-59) or Cerenkov-measurement (P-32), respectively. The activities recovered from lung homogenates rendered is possible to calculate the received amount of bacteria. 40-80% of germs would be recovered in the lung after application by intratracheal injection while only 1-7% could be recovered after individual aerosol application lasting for 15-25 minutes.

Aerosols↗

Bacteria-immune system interactions. IV. Effect of various treatments on the binding of bacteria to human lymphocytes.

Pretreatment of lymphocytes with selected saccharides inhibited the binding of Bacillus globigii (B. globigii) to lymphocytes but had no effect on Escherichia coli 2 (E. coli 2) adherence. The binding of both bacteria was prevented by lymphocyte pretreatment with trypsin or pronase. Periodate treatment of lymphocytes inhibited the binding of E. coli 2 but not of B. globigii. This inhibition could be reversed by borohydrate treatment of lymphocytes. Trypsin, pronase or periodate treatment of bacteria had no effect on the binding to lymphocytes. The data obtained suggest that: (a) E. coli 2 and B. globigii have different mechanisms of binding to lymphocytes; and (b) the lymphocyte receptor for E. coli 2 is of a glycoprotein nature.

Adhesiveness↗

[Qualitative and quantitative examination of bacteria found in aquatic habitats. 2. Communication: application of miniturizid multitest systems for identification and biochemical typing of bacteria using a multi-point method (author's transl)].

A large number of bacteria were characterized and identified using a miniturized multi-test system and an automatic inoculator. This testsystem is suitable for ecological studies to get information about the qualitative and quantitative composition of bacterial populations. More than that a biochemical typing of bacteria is possible.

Bacteria↗