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PubMed1996

The effects of magainin 2, cecropin, mastoparan and melittin on Brucella abortus.

The effect of the alpha-helical polycationic peptides magainin 2, melittin, mastoparan and cecropin on the viability of Brucella abortus 544 (type species), B. abortus S19 (vaccine strain) and B. abortus S2308 (vaccine challenge strain) was determined. Rough mutants of these strains and the rough candidate vaccine strain B. abortus RB51 were also tested. S. typhimurium was used as a control. The peptides did not affect the viability of B. abortus smooth strains but some of the peptides affected viability of the rough strains. Magainin 2 at a concentration of 100 micrograms ml-1 did not reduce the viability of the rough B. abortus strains. Cecropin at a concentration of 15 micrograms ml-1 reduced the viability of the rough strains by approximately 10-fold. Mastoparan at a concentration of 50 micrograms ml-1 reduced the viability of the rough strains by approximately 100-fold. Melittin at a concentration of 20 micrograms ml-1 reduced the viability of the rough strains of B. abortus by approximately 1000-fold. The brucellae were significantly more resistant to all the cationic peptides than was S. typhimurium.

Anti-Bacterial Agents↗

PubMed1999

Drug-antibody conjugates with anti-HIV activity.

Human immunodeficiency virus (HIV)-specific peptide antibody-brefeldin A conjugates and antibody-glaucarubolone conjugates directed to cell surface viral glycoprotein epitopes were prepared and tested for antiviral activity. A selective response was observed both on survival of cell lines permanently infected with lentiviruses and on HIV infectivity. With human peripheral blood mononuclear cells (PBMCs), the conjugate also was effective in reducing virus titers. The effectiveness of an HIV-specific peptide antibody-brefeldin A conjugate was enhanced by combination with 3'-azido-3'-deoxythymidine (AZT) and was effective against AZT-resistant isolates in combination with AZT. The conjugates reduced virus production in MOLT-4 cells and in HIV-1-infected PBMCs without affecting the viability of uninfected cells.

Amino Acid Sequence↗

PubMed2006

Induction of cytokine expression by herpes simplex virus in human monocyte-derived macrophages and dendritic cells is dependent on virus replication and is counteracted by ICP27 targeting NF-kappaB and IRF-3.

Macrophages and dendritic cells (DCs) play essential roles in host defence against microbial infections. In the present study, it is shown that human monocyte-derived macrophages and DCs express both type I and type III interferons (IFNs) [IFN-alpha, IFN-beta and interleukin 28 (IL-28), IL-29, respectively], tumour necrosis factor alpha and the chemokines CCL5 and CXCL10 after herpes simplex virus 1 (HSV-1) infection. The cytokine-inducing activity of HSV-1 was dependent on viability of the virus, because UV-inactivated virus did not induce a cytokine response. Pretreatment of the cells with IFN-alpha or IL-29 strongly enhanced the HSV-1-induced cytokine response. Both IFN-alpha and IL-29 decreased viral immediate-early (IE) gene infected-cell protein 27 (ICP27) transcription, suggesting that IL-29 possesses antiviral activity against HSV-1 comparable to that of IFN-alpha. Macrophage infection with HSV-1 lacking functional ICP27 (d27-1 virus) resulted in strongly enhanced cytokine mRNA expression and protein production. In contrast, viruses lacking functional IE genes ICP0 and ICP4 induced cytokine responses comparable to those of the wild-type viruses. The activation of transcription factors IRF-3 and NF-kappaB was strongly augmented when macrophages were infected with the ICP27 mutant virus. Altogether, the results demonstrate that HSV-1 both induces and inhibits the antiviral response in human cells and that the type III IFN IL-29, together with IFN-alpha, amplifies the antiviral response against the virus. It is further identified that viral IE-gene expression interferes with the antiviral response in human macrophages and ICP27 is identified as an important viral protein counteracting the early innate immune response.

Cytokines↗

PubMed1984

Regrowth of aminoglycoside-resistant variants and its possible implication for determination of MICs.

Regrowth of aminoglycoside-resistant variants was seen when large inocula of two strains of Escherichia coli were incubated with gentamicin in concentrations well above their MICs (0.5 micrograms/ml). The extent of the selection of resistant variants was proportional to the concentration of gentamicin during incubation; after incubation with gentamicin (greater than or equal to 2 micrograms/ml for 24 h), all bacteria were resistant to at least 8 micrograms/ml. Bacteria resistant to these concentrations always formed small colonies, whereas variants resistant to lower concentrations (1 to 2 micrograms/ml) could form both small and normal colonies. The regrowth of resistant variants could be monitored by luciferase assay of intracellular ATP in cultures incubated with gentamicin (less than or equal to 2 micrograms/ml). In cultures incubated with higher concentrations, regrowth did occur, although this did not result in viability (CFU per milliliter) or ATP levels above those of the initial inocula. The implications of this regrowth for MIC determinations in broth and the possible clinical revelance of the resistant variants are discussed.

Aminoglycosides↗

PubMed2000

Viability of probiotic (Bifidobacterium, Lactobacillus acidophilus and Lactobacillus casei) and nonprobiotic microflora in Argentinian Fresco cheese.

We evaluated the suitability of Argentinian Fresco cheese as a food carrier of probiotic cultures. We used cultures of Bifidobacterium bifidum (two strains), Bifidobacterium longum (two strains), Bifidobacterium sp. (one strain), Lactobacillus acidophilus (two strains), and Lactobacillus casei (two strains) in different combinations, as probiotic adjuncts. Probiotic, lactic starter (Lactococcus lactis and Streptococcus thermophilus), and contaminant (coliforms, yeasts, and molds) organisms were counted at 0, 30, and 60 d of refrigerated storage. Furthermore, the acid resistance of probiotic and starter bacteria was determined from hydrochloric solutions (pH 2 and 3) of Fresco cheese. The results showed that nine different combinations of bifidobacteria and L. acidophilus had a satisfactory viability (count decreases in 60 d <1 log order) in the cheese. Both combinations of bifidobacteria and L. casei cultures assayed also showed a satisfactory survival (counts decreased <1 log order for bifidobacteria but no decrease was detected for L. casei). On the other hand, the three combinations of bifidobacteria, L. acidophilus, and L. casei tested adapted well to the Fresco cheese environment. When a cheese homogenate at pH 3 was used to partially simulate the acidic conditions in the stomach, the probiotic cultures had an excellent ability to remain viable up to 3 h. At pH 2, the cell viability was more affected; B. bifidum was the most resistant organism. This study showed that the Argentinian Fresco cheese could be used as an adequate carrier of probiotic bacteria.

Bifidobacterium↗

PubMed2003

Immune-enhancement effect of the herbal combination Allergina.

BACKGROUND: The herbal formulation, Allergina, has long been used for various diseases. It is known to have an anti-microbial and anti-virus activity. However, it is still unclear how Allergina has these effects in experimental models. We investigated the effect of Allergina on the proliferation of T cell and production of cytokines in human T-cell line, MOLT-4 cells, and mouse peritoneal macrophages. METHODS: The MOLT-4 cells were cultured for 24 h in the presence or absence of Allergina. Allergina significantly increased the cell viability by 26.9+/-5.4% (P<0.05) and interleukin (IL)-2, IL-4 and interferon (IFN)-gamma production compared with media control (about 4-fold for IL-2, 2.5-fold for IL-4 and 3.4-fold for IFN-gamma, P<0.05). Maximal effective concentration of Allergina was 1 mg/ml for IL-2 and, 0.01 mg/ml for IL-4 and IFN-gamma. Allergina alone or Allergina plus recombinant IFN-gamma (rIFN-gamma) increased the production of tumor necrosis factor (TNF)-alpha, but Allergina decreased the production of TNF-alpha on rIFN-gamma plus LPS-stimulated macrophages. In addition, Allergina increased the production of IL-12 on mouse peritoneal macrophages and peripheral blood mononuclear cells. CONCLUSION: Allergina may have an immune-enhancement effect through the cytokine production.

Adjuvants, Immunologic↗

PubMed2004

Inhibition of Salmonella enterica serovars by microcin J25.

Escherichia coli microcin J25 (MccJ25) is a 2107-Da peptide antibiotic whose uptake into E. coli is mediated by the outer-membrane receptor FhuA and the inner membrane proteins TonB, ExbB, ExbD, and SbmA. A survey of the sensitivity of several Salmonella enterica serovars showed that the antibiotic was highly active against some serovars, while S. Typhimurium, S. Derby, and some S. Enteritidis strains were completely resistant. Resistant strains became hypersensitive to MccJ25 when given the fhuA gene of E. coli, indicating that insensitivity is due to the inability of the FhuA protein to mediate penetration of MccJ25. Whereas in E. coli MccJ25 targets RNA polymerase, in S. Typhimurium it inhibits not only RNA synthesis but also cell respiration. Fluorescence viability staining showed that S. Typhimurium cells exposed to MccJ25 remain viable but are unable to form colonies.

Anti-Bacterial Agents↗

PubMed2004

Low concentrations of the non-ionic detergent Nonidet P-40 interfere with sterol biogenesis and viability of the yeast Saccharomyces cerevisiae.

Mild non-ionic detergents are used for solubilization of hydrophobic substrates in yeast growth media at concentrations 0.1-1%. Our data show that low concentrations of Nonidet P-40 may significantly affect lipid biogenesis in the yeast Saccharomyces cerevisiae. The uptake and esterification of external [4-14C]-cholesterol is strongly reduced in hem1 mutants treated with low concentrations of Nonidet P-40. Significant inhibitory effect of NP-40 on sterol uptake and esterification was evident both in non-growing and growing cells supplemented with external cholesterol. Increased levels of sterol precursors (squalene, lanosterol) in hem1 cells grown in complex medium with cholesterol indicated general interference of NP-40 with sterol biosynthesis. NP-40 in the growth medium affected also cell viability estimated as the colony forming ability. More attention should be therefore paid to possible effects of mild detergents at low concentrations generally considered to be harmless, especially in cells with disturbed lipid biogenesis.

Biological Transport↗

PubMed1999

The viability of bifidobacteria introduced into kimchi.

The viability of bifidobacteria in mul-kimchi, a type of kimchi with added water, was investigated under various conditions. When a mul-kimchi preparation was inoculated with five strains of Bifidobacterium at a concentration of 10(7) cfu ml-1, Bif. longum JK-2 showed the highest viability, maintaining a population of 10(6) cfu ml-1 after 1 week at 4 degrees C. The influence of NaCl concentration and initial pH on viability was further investigated in mul-kimchi inoculated with Bif. longum JK-2; NaCl concentrations greater than 3% (w/w) reduced viability considerably. In kimchi started with an initial pH of 6.5, the cells showed the highest survival. When mul-kimchi containing 2% NaCl (w/w) was inoculated with 10(8) cfu ml-1 Bif. longum JK-2, there was a 10-fold reduction in viability during 10 d of incubation at 4 degrees C. These results demonstrate acceptable levels of the organism in the product, suggesting the possible use of selected strains of bifidobacteria in commercial kimchi production.

Bifidobacterium↗

PubMed2003

Additive antimicrobial activity of calcium hydroxide and chlorhexidine on common endodontic bacterial pathogens.

It is an established procedure to use calcium hydroxide [Ca(OH)2] as a therapeutic component to achieve sterilization of infected root canals. Unfortunately, some bacterial species are relatively resistant to Ca(OH)2 and are therefore associated with treatment failures. The objective of this study was to identify combinations of substances that improve antimicrobial activity. Ca(OH)2 was used as a suspension or as a component of gutta-percha points. Efficacy of killing was compared for (a) Ca(OH)2 suspension alone; (b) zinc oxide (ZnO) points alone; (c) Ca(OH)2 suspension combined with ZnO points; (d) Ca(OH)2 suspension combined with ZnO/chlorhexidine points; and (e) Ca(OH)2 points combined with ZnO/chlorhexidine points. Common endodontopathogenic bacterial species (Enterococcus faecalis, Fusobacterium nucleatum, Peptostreptococcus micros, Porphyromonas gingivalis, Streptococcus intermedius) in pure cultures served as target organisms. In the in vitro assay, the antibacterial substances and approximately 10(7) colony forming units of test bacteria were co-incubated in diluted human serum under growth conditions appropriate for the single bacterial species. Samples were taken on days 0, 1, 2, 4, 7, and 14 of incubation and viable counts determined. Gram-negative bacteria were sufficiently killed by Ca(OH)2 alone. Combinations with other agents did not improve the killing kinetics. For the inactivation of the Gram-positive bacteria P. micros and S. intermedius, a combination of Ca(OH)2 and ZnO/chlorhexidine, killed the bacteria faster than Ca(OH)2 alone. For E. faecalis, combining Ca(OH)2 suspension with ZnO/chlorhexidine points lead to a faster decrease in the overall number of viable bacteria, but no test conditions lead to the complete loss of culture viability. The results of this study support the use of a combination of Ca(OH)2 and chlorhexidine for faster eradication of P. micros and S. intermedius from infected root canals.

Anti-Infective Agents, Local↗

PubMed2004

Comparative survival of probiotic lactobacilli spray-dried in the presence of prebiotic substances.

AIMS: Probiotic milk-based formulations were spray-dried with various combinations of prebiotic substances in an effort to generate synbiotic powder products. METHODS AND RESULTS: To examine the effect of growth phase and inclusion of a prebiotic substance in the feed media on probiotic viability during spray-drying, Lactobacillus rhamnosus GG was spray-dried in lag, early log and stationary phases of growth in reconstituted skim milk (RSM) (20% w/v) or RSM (10% w/v), polydextrose (PD) (10% w/v) mixture at an outlet temperature of 85-90 degrees C. Stationary phase cultures survived best (31-50%) in both feed media and were the most stable during powder storage at 4-37 degrees C over 8 weeks, with 30-140-fold reductions in cell viability at 37 degrees C in RSM and PD/RSM powders, respectively. Stationary phase Lact. rhamnosus GG was subsequently spray-dried in the presence of the prebiotic inulin in the feed media, composed of RSM (10% w/v) and inulin (10% w/v), and survival following spray-drying was of the order 7.1-43%, while viability losses of 20,000-90,000-fold occurred in these powders after 8 weeks' storage at 37 degrees C. Survival of the Lactobacillus culture after spray-drying in powders produced using PD (20% w/v) or inulin (20% w/v) as the feed media was only 0.011-0.45%. To compare different probiotic lactobacilli during spray-drying, stationary phase Lact. rhamnosus E800 and Lact. salivarius UCC 500 were spray-dried using the same parameters as for Lact. rhamnosus GG in either RSM (20% w/v) or RSM (10% w/v) and PD (10% w/v). Lact. rhamnosus E800 experienced approx. 25-41% survival, yielding powders containing approximately 10(9) CFU g(-1), while Lact. salivarius UCC 500 performed poorly, experiencing over 99% loss in viability during spray-drying in both feed media. In addition to the superior survival of Lact. rhamnosus GG after spray-drying, both strains experienced higher viability losses (570-700-fold) during storage at 37 degrees C over 8 weeks compared with Lact. rhamnosus GG. CONCLUSIONS: Stationary phase cultures were most suitable for the spray-drying process, while lag phase was most susceptible. The presence of the prebiotics PD and inulin did not enhance viability during spray-drying or powder storage. SIGNIFICANCE AND IMPACT OF THE STUDY: High viability (approximately 10(9) CFU g(-1)) powders containing probiotic lactobacilli in combination with prebiotics were developed, which may be useful as functional food ingredients for the manufacture of probiotic foods.

Animals↗

PubMed1968

Synthesis of protein and ribonucleic acid in a psychrophile at normal and restrictive growth temperatures.

A defined medium was capable of supporting the growth of a psychrophilic coccus over its growth temperature range, -4 to 25 C. A rapid loss of viability occurred when exponential cells were transferred to growth-restricting temperatures above 25 C. Comparative studies of the chemistry of exponential-phase cells and cells exposed to supermaximum temperature indicated that this loss of viability is not due to temperature-induced membrane damage, inhibition of respiration or energy metabolism, or depletion of intracellular reserves. Moribund and dead cell populations showed an elevated level of intracellular adenosine-5'-triphosphate and amino acids-a finding reflected in the reduced rate of amino acid synthesis during the recovery of heat-shocked cells-and also leakage of degraded ribonucleic acid products into the medium. Incorporation studies indicated that loss of viability at 30 C was correlated with inhibition of protein synthesis, followed later by inhibition of ribonucleic acid synthesis. Deoxyribonucleic acid synthesis was unaffected by temperature above the maximum.

Acetates↗

PubMed2000

Characterization of high density monolayers of the biofilm bacterium Caulobacter crescentus: evaluating prospects for developing immobilized cell bioreactors.

Caulobacters are biofilm-forming members of the natural flora of soil and aquatic environments, which exhibit several characteristics that make them attractive for development of high surface area microbial bioreactors or biosensors. Although caulobacters are well characterized genetically, little is known about their biofilm-forming characteristics as a monoculture, or their tolerance of bioreactor-like conditions. Here we investigated the ability of caulobacters to spontaneously form high-density monolayers on artificial surfaces under a variety of environmental conditions, using phase contrast image analysis to assess biofilm density, and epifluorescence with the vital stain DiBAC to assess viability. With adequate nutrition, extremely dense monolayers formed within 24-48 h, and maintained near 100% viability in experiments ranging up to 22 days. When areas were abraded to remove cells, repopulation occurred rapidly with characteristics similar to the population of a clean surface. When established monolayers were starved for nutrients, a significant fraction of the cells detached from the surface, and cells remaining on the surface no longer tested as viable. Within 4-6 h of nutrient restoration, however, cells in the monolayer again appeared normal and tested as 100% viable. This is the first demonstration that Caulobacter crescentus is stable and amenable to high density monolayer growth and resists starvation, though some cells may express a programmed response to detach from the surface under severe nutrient limitation.

Barbiturates↗

PubMed1975

DNA release as a direct measure of microbial killing. I. Serum bactericidal activity.

A new method for quantification of microbial killing is presented, based upon the assumption that release of DNA from an organism can be taken as direct evidence of cell death. The assay was applied to measurement of serum bactericidal activity. A "serum-sensitive" Escherichia coli, whose DNA was pre-labeled with [14C] thymidine, released 95 to 100% of its radioactivity upon exposure to human serum for 120 min. This was accompanied by a fall in viability with less than 0.1% of the bacteria surviving. Heating serum to 56 degrees C for 30 min completely abolished both DNA release and killing. Normal serum did not release DNA from a "serum-resistant". Salmonella typhimurium whereas exposure to ampicillin caused both significant killing and DNA release. This assay is highly specific, sensitive, and rapid.

Ampicillin↗

PubMedDate not supplied

Preconditioning for moisture control in ethylene oxide sterilization.

The moisture condition of a product must be controlled to obtain sterility and maintain product viability during ethylene oxide sterilization. A sterilization process in which prehumidification of product is used to provide moisture condition control was shown to be effective in 2 years of production sterilization experience. On-stream process analysis with a gas chromatograph showed that addition of steam to the chamber was unnecessary when properly preconditioned medical devices packaged in corrugated paper-board were sterilized. Reproducible gas-chamber moisture control and microbial sterility assurance were obtained.

Catheters, Indwelling↗

PubMed2000

IB-367, a protegrin peptide with in vitro and in vivo activities against the microflora associated with oral mucositis.

Although the microflora associated with oral mucositis initiated by cytotoxic therapy is not well characterized, several studies suggest that reduction of the microbial load in the oral cavity has some clinical benefit. The MICs of IB-367, a synthetic protegrin analog, ranged from 0.13 to 64 microgram/ml for gram-positive bacteria (Streptococcus mitis, Streptococcus sanguis, Streptococcus salivarius, and Staphylococcus aureus) and from 0.06 to 8 microgram/ml for gram-negative species (Klebsiella, Escherichia, and Pseudomonas). IB-367 exhibited rapid, microbicidal activity against both log- and stationary-phase cultures of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. At concentrations near the MICs for these two organisms (4 and 2 microgram/ml, respectively), IB-367 reduced viability by more than 3 logs in less than 16 min. Similarly, IB-367 effected a 4-log reduction of the endogenous microflora in pooled human saliva within 2 min at 250 microgram/ml, a concentration readily attained by local delivery. After nine serial transfers at 0.5x the MIC, the MIC of IB-367 for MRSA and P. aeruginosa increased only two to four times. In a phase I clinical study with healthy volunteers, IB-367 was well tolerated, with no detectable systemic absorption. One hour after treatment with 9 mg of IB-367, the prevalence of gram-negative bacteria and yeast was reduced, and the density of the predominant gram-positive oral flora was decreased 1,000 times. IB-367's properties (speed of killing, breadth of spectrum, and lack of resistance) make the compound a strong candidate for the prophylaxis of oral mucositis. Phase II clinical trials with IB-367 are under way for this indication in immunocompromised subjects.

Anti-Bacterial Agents↗

PubMed1998

Disinfectant testing: use of the Bioscreen Microbiological Growth Analyser for laboratory biocide screening.

A new method is described for screening potential biocides based on the traditional suspension test using the Bioscreen optical plate reader. This new method is rapid, reproducible, quantitative and cost effective. Data obtained by this new method are not directly equivalent to the log reduction normally quoted, but give a measurement of the total effect of the biocide on the microbe population, measuring the effect of injury as well as death (non-viability). The method allows for the routine examination of disinfection kinetics, the study of which leads to greater scientific insight into disinfection than that achieved by the standard 5 min, one-point, disinfection tests currently employed.

Colony Count, Microbial↗

PubMed2001

Synergic activity of D-cycloserine and beta-chloro-D-alanine against Mycobacterium tuberculosis.

D-Cycloserine (DCS) is a peptidoglycan inhibitor. Although very effective against Mycobacterium tuberculosis, it is seldom employed in the management of this infection due to its high toxicity. beta-Chloro-D-alanine (another peptidoglycan inhibitor) reduces the MIC of DCS from 50 to 2.5 mg/L at a concentration significantly below its MIC for this organism. A reduction in bacterial viability and significant growth inhibition (as quantified by the X/Y quotient for the Bactec radiometric procedure) were observed with subinhibitory concentrations of both drugs. It is suggested that this powerful synergic effect should be the object of in vivo and eventually clinical trials.

Cycloserine↗