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A method for tracking the locomotion of an isolated microorganism in real time.

An inexpensive, real-time recording system ("bug-tracker") for tracking the movements of an isolated microorganism was assembled using a close-up video system, a video memory card, and a personal computer. An isolated organism moving in an almost two-dimensional plane is viewed by the close-up video camera, and a selected video frame is digitized by the video memory card into 256 x 256 pixels (picture points). The pixels of the ith frame are subtracted from those of the i-lth frame to delete images other than that of the image of the moving organism. Windows with an optimized size are generated commonly in the ith and i-lth frame to reduce the number of pixels directly accessed by the computer, and the address of the pixel with the largest value inside the windows identifies the coordinates of the organism on the X and Y axes. By optimizing the size of a window for a given organism, the sampling for the X and Y coordinates can be made at times separated by approximately 0.2 to 2.0 seconds. Data are automatically filed on a floppy disk.

Animals↗

Anthranilate synthase in microorganisms and plants.

The enzymology of anthranilate synthase (EC 5.4.99.6) in microorganisms and plants is reviewed. Aminoacid sequences of the enzyme subunits in different species are compared, and the mechanism of reaction is discussed.

Amino Acid Sequence↗

Isolation and biological activities of signal transduction inhibitors from microorganisms and plants.

Microorganisms and plants are treasuries of secondary metabolites having unique structures and biological activities. We have isolated inhibitors of cellular signal transduction from them. Phosphatidylinositol turnover inhibitors include inostamycin, piericidins and echiguanines. Erbstatin and lavendustin A were isolated as tyrosine kinase inhibitors, and dephostatin was isolated as a tyrosine phosphatase inhibitor. Recently, we have isolated a new vinca alkaloid having anti-ras activity. These signal transduction inhibitors will be useful for mechanistic studies and suppression of cancer.

1-Phosphatidylinositol 4-Kinase↗

Inhibition of experimental metastasis by enzyme inhibitors from microorganisms and plants.

Various antibacterial compounds, antitumor compounds, enzyme inhibitors and recent signal transduction inhibitors have been discovered from microorganisms and plants. Therefore, it should be possible to find antimetastatic compounds from these sources, if a simple assay system is available. We isolated several enzyme inhibitors from nature to inhibit experimental metastasis. Leupeptin is an old protease inhibitor and inhibited blood-borne lung metastasis of hepatoma cells in rats. A leupeptin analogue inhibiting urokinase inhibited in vitro invasion of human fibrosarcoma cells. Alpha-glucosidase inhibitors such as epi-CPL and baicalein inhibited in vitro invasion and in vivo metastasis of mouse melanoma cells. A mannosidase inhibitor, mannostatin A, also inhibited in vitro invasion of mouse melanoma cells. Oncogene function inhibitors induce normal phenotypes in the oncogene-expressing cells. As expected, they inhibited tumor cell invasion in vitro.

Animals↗

Dietary marine lipid does not alter susceptibility of (NZBxNZW)F1 mice to pathogenic microorganisms.

Female (NZBxNZW)F1 mice were fed for one month with a diet in which the sources of fat were either melted beef tallow or fish oil, the latter regimen being associated with a marked decrease in the expression of auto-immune disease in these animals. To test whether or not this beneficial effect was associated with an increased risk of infection, animals fed each of these diets were challenged intraperitoneally with graded doses of four different classes of microorganisms: a gram positive bacillus that is an intracellular parasite (Listeria monocytogenes), an exotoxin-producing gram negative bacillus (Pseudomonas aeruginosa), a yeast (Candida albicans), and a herpes group virus (murine cytomegalovirus). There was no difference in the susceptibility of the animals fed the two different diets to any of these infections as shown by either determinations of the LD50 for each organism or by assessment of the times of survival for those animals that did succumb. We conclude that the beneficial anti-inflammatory effects of fish oil diets in these mice are not associated with an increased risk of infection with a variety of microbial agents.

Animals↗

Transitory acidification of semen as a potential method for the inactivation of some pathogenic microorganisms. Effect on fertilization and development of ova in superovulated heifers.

The fertilizing capacity of transitorily acidified semen was investigated with respect to using acidification as a method for destroying or inactivating acid labile pathogenic microorganisms in semen prior to freezing. Ejaculates diluted 1:1 with phosphate buffered saline (PBS) were acidified to pH 5.0 for 2 or 5 minutes before being returned to their original pH. They were then frozen by commercial methods and used to artificially inseminate superovulated heifers. A total 739 ova and embryos was collected from 119 inseminated donors. The fertilization rate was 88%, and 78% of the embryos were of transferable quality. Semen acidification had no apparent effect on the post-thaw percentage of intact acrosomes.

Journal Article↗

Esterases of xylan-degrading microorganisms: production, properties, and significance.

This review focuses on the description of recently discovered esterase enzymes involved in xylan degradation (acetyl xylan, feruloyl, and p-coumaroyl esterases). The occurrence of these enzymes in various microorganisms, assays used for determination of their activity, induction and production on different substrates, interaction with other xylanolytic enzymes, mode of action, substrate specificity, and biochemical characteristics are presented. The nature of substrates on which acetyl xylan esterase, feruloyl, and p-coumaroyl esterase are active and their role in xylan hydrolysis is emphasized. The potential applications of xylan-debranching esterases are outlined and their significance to applied microbiology is discussed.

Acetylesterase↗

Design and analysis of experiments on mutagenicity. II. Assays involving microorganisms.

The design and statistical analysis of mutagenicity experiments involving microorganisms and a single dose of mutagen are discussed. Test statistics are derived for use in determining the mutagenicity of a chemical when survival data are available and also when such data are not available. One's likelihood (power) of correctly concluding a chemical is mutagenic is examined, and minimum total sample sizes required for 95% power are presented. It is found that one generally has greater power when survival data are available. Required precision is estimating survival is discussed in reference to type-1 and type-2 errors. The proper use of the formulae and figures presented is illustrated by examples.

Bacteria↗

Antimutagenicity of cell fractions of microorganisms on potent mutagenic pyrolysates.

The inactivation of 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 2-amino-6-methyldipyrido[1,2-a: 3',2'-d]imidazole (Glu-P-1) by binding of mutagenic pyrolysate to fractions of microorganisms (their desmutagenic and bio-antimutagenic activity) was investigated. All strains bound Trp-P-1 effectively, but Glu-P-1 to a lesser extent. The Gram-negative bacteria (GNB) could bind about 10-20 micrograms/mg of Trp-P-1 more than the Gram-positive bacteria (GPB), and about 50-60 micrograms/mg more than the yeasts. The cell wall skeletons of all strains tested had great binding ability, but in the cytoplasm of all strains tested it was lower. The peptidoglycan, outer membrane, and glucan isolate from the cell wall skeletons showed the highest binding ability to Trp-P-1. The cell wall skeletons of the tested strains greatly inhibited the mutagenicity induced by Trp-P-1, and to a lower extent that of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx). Although the cultured broth and solution of cells extracted by phosphate buffer (pH 7.0) showed antimutagenicity against Trp-P-1, this activity was lower than the binding of Trp-P-1 to the cells. The cultured broth and freeze-dried cytoplasms of yeast cells showed bio-antimutagenicity towards Trp-P-1, but those of all bacteria tested did not.

Amines↗

Specificities of extracellular and ribosomal serine proteinases from Bacillus natto, a food microorganism.

The specificities of extracellular and ribosomal serine proteinase from Bacillus natto, a food microorganism, were investigated. Both proteins have highly restricted and characteristic specificities. With the extracellular serine proteinase, initial cleavage site was observed at Leu15-Tyr16, secondary site at Ser9-His10 and additional cleavage sites at Gln4-His5 and His5-Leu6 in the oxidized insulin B-chain. Hydrolysis of proangiotensin with the extracellular serine proteinase was observed primarily at Phe8-His9 and secondary at Tyr4-Ile5. The extracellular serine proteinase has a Km of 0.08 mM and kcat of 3 S-1 for angiotensin hydrolysis. With the ribosomal proteinase, initial cleavage site of the oxidized insulin B-chain was observed at Leu15-Tyr16 and additional cleavage site at Phe24-Phe25. Hydrolysis of proangiotensin was observed at Tyr4-Ile5 bond with the ribosomal proteinase.

Angiotensinogen↗

Isolating and characterizing deep-sea marine microorganisms.

We have isolated several microorganisms that are adapted to living in the extremes of the deep-sea environment. They include barophilic bacteria, which are able to grow at high hydrostatic pressure, but that are unable to grow at atmospheric pressure, and organic-solvent-tolerant bacteria, which are able to grow in the presence of toxic organic solvents such as toluene or benzene. In this review, we describe how to isolate such extremophiles, and we outline the characteristics of several strains that have been recovered from the deep-sea environment.

Bacteria↗

Modelling the growth, survival and death of microorganisms in foods: the UK food micromodel approach.

Techniques for the development of mathematical models in the area of predictive microbiology have greatly improved recently, allowing better and more accurate descriptions of microbial responses to particular environmental conditions, thus enabling predictions of those responses to be made with greater confidence. Recognising the potential value of applying these techniques in the food industry, the Ministry of Agriculture, Fisheries and Food (MAFF) initiated a nationally coordinated five-year programme of research into the growth and survival of microorganisms in foods, with the aim of developing a computerised Predictive Microbiology Database in the UK. This initiative has resulted in the systematic generation of data, through protocols which ensure consistency of methodology, so that data in the database are truly comparable and compatible, and lead to reliable predictive models. The approaches taken by scientists involved in this programme are described and the various stages in the development of mathematical models summarized. It is hoped that this initiative and others being developed in the USA, Australia, Canada and other countries, will encourage a more integrated approach to food safety which will influence all stages of food production and, eventually, result in the development of an International Predictive Microbiology Database.

Bacteria↗

Characteristics of a multicomponent first-order model for thermal inactivation of microorganisms and enzymes.

There are several mathematical models that express different types of non-linear thermal inactivation of microorganisms and enzymes. Among the models the characteristics of a multicomponent first-order model were studied on a calculative simulation. For the initial period of inactivation the pattern of the model was predominantly affected by the kinetics of the component with the highest initial concentration. For longer periods of inactivation the pattern was predominantly affected by the kinetics of the component whose rate constant was the lowest. It was also found that the observation period affected the apparent pattern of the model. The estimation method of parameter values of the model from inactivation curves was then studied.

Enzymes↗

Novel bioemulsifiers from microorganisms for use in foods.

The main objective of this study was to test the range of microorganisms for production of extracellular, high molecular weight emulsifiers for potential use in foods. A standard emulsification assay developed specifically for assessing food emulsifiers was used to examine 24 extracellular microbial products from bacteria, yeasts and algae. Of the 24 products tested, nine had emulsification ability that was as good as and eight had emulsifying properties that were better than those of the commonly used food emulsifiers gum arabic and carboxymethylcellulose. The eight good producer organisms included the yeasts Candida utilis, Candida valida, Hansenula anomala, Rhodospiridium diobovatum and Rhodotorula graminis, the red alga Porphiridium cruentum, and the bacteria Klebsiella spp. and Acinetobacter calcoaceticus. Of these, C. utilis was selected for further study due to the excellent emulsification properties of its extracellular products and food-grade status of the organism. Crude preparations of the bioemulsifier from C. utilis exhibited low viscosity and had a carbohydrate content of over 80%. Preliminary trials showed that the bioemulsifier from this organism had potential for use in salad cream.

Acinetobacter calcoaceticus↗

Low molecular weight immunomodulators produced by microorganisms.

In order to develop a new class of low molecular weight immunomodulators for the treatment of incurable diseases involving cancer, hematologic diseases and inflammation, we have sought cytokine inducers and inhibitors of cell adhesion to the extracellular matrix among the low molecular compounds produced by microorganisms. Cytogenin, cytoblastin (a monokine inducer), conagenin (a lymphokine inducer) and delaminomycins and IC-101 (inhibitors of cell adhesion to extracellular matrices) have been recently found in our institute. In this report, we describe the principles of screening and structures, and discuss their biological activities.

Actinobacteria↗

An epidemiological study assessing the relative importance of airborne and direct contact transmission of microorganisms in a medical intensive care unit.

A prospective epidemiological survey was carried out over a period of seven weeks in a medical intensive care unit. Bacteria from patients, staff and air were monitored and the transmission of isolated microorganisms was followed. Handwashing samples revealed pathogenic bacteria in 30.8% of physicians (average number of colony forming units: 71,300 per hand) and 16.6% of nurses (39,800 cfu per hand). Air cultures yielded pathogens in 15% of sampling periods and nine of 53 patients were found to be colonized with Gram-negative bacteria, Staphylococcus aureus or Candida spp. The spectrum of bacteria recovered from patients and air was generally different, whereas strains recovered from patients and their attendants' hands were indistinguishable on multiple occasions. The results of this study confirm that direct contact is the principal pathway of microbial transmission, whereas little evidence for a significant role of airborne transmission is shown. The call for more extensive air-filtering and ventilation systems in medical intensive care units is not supported by the results shown in this communication.

Air Microbiology↗

Bowel microorganisms--a target for selective antimicrobial control.

This article reviews the eight factors that determine the outcome of selective antimicrobial control (SAC) a technique aimed at the clearance of intestinal Gram-negative bacillary carriage by means of lethal faecal anti-microbial concentrations. They are as follows: (i) the carrier state; (ii) compliance; (iii) SAC aiming at prophylaxis vs treatment; (iv) minimum bactericidal concentration (MBC) of the antimicrobial; (v) dosage; (vi) pharmacokinetics; (vii) faecal inactivation; and (viii) microorganisms to be controlled. In the second part, non-absorbable SAC regimens are compared with absorbable trimethoprim/sulphamethoxazole (TMP/SMZ) and the fluoroquinolones in different clinical settings including neutropenia, intensive care, hepatic encephalopathy, liver transplantation and the salmonella carrier state. Ablation of gut carriage and superinfections are the main endpoints reviewed in this article. The newer fluoroquinolones are potent SAC agents to deal with enterobacteria. Pseudomonads are the major gap in their SAC spectrum. TMP/SMZ emerges as a SAC agent of limited value, whilst the newer non-absorbable combination of polymyxin/tobramycin seems to be the most potent SAC programme since it has activity against pseudomonads. In a third part, three current issues--the emergence of resistance, the selectivity and the tissue effect are discussed. Finally, a potent fluoroquinolone combined with oral polymyxin/tobramycin seems to be the most effective SAC programme currently available to control enterobacteria and pseudomonads in patients in whom bacterial translocation is a risk with minimal risk of resistance emerging.

4-Quinolones↗