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PubMed1989

The expression of the superoxide dismutase gene in Halobacterium cutirubrum and Halobacterium volcanii.

The gene encoding the Mn-containing superoxide dismutase (SOD) from Halobacterium cutirubrum has been cloned and sequenced. The deduced amino acid sequence is homologous to the sequences of Fe and Mn SODs from eubacteria. The high degree of amino acid identity between the archaebacterial and eubacterial proteins suggests that a SOD gene may have been laterally transferred between eubacteria and archaebacteria sometime after the accumulation of atmospheric oxygen. Consensus elements of halobacterial promoters are found upstream of the coding region, however, the spacing between them and the transcription start site is greater than in other genes. Termination of transcription occurs in five consecutive T residues that are preceded by a GC-rich sequence that has short inverted repeats. In addition to the authentic SOD gene, H. cutirubrum also contains a putative pseudogene. The SOD levels and growth rates of H. cutirubrum and Halobacterium volcanii were tested in response to treatment by paraquat, an intracellular generator of superoxide. In H. volcanii the growth rate slowed, and SOD was strongly induced throughout prolonged treatment with paraquat. In H. cutirubrum the same effects were noticed initially, but after 48 h exposure to the drug, the growth rate increased and the SOD level decreased. Production of paraquat resistant mutants of H. cutirubrum may play a part in this process, however, some type of physiological adaptation is also probably required.

Amino Acid Sequence↗
PubMed2001

The immune response to ocular herpes simplex virus type 1 infection.

Herpes simplex virus type 1 (HSV-1) is a prevalent microbial pathogen infecting 60% to 90% of the adult world population. The co-evolution of the virus with humans is due, in part, to adaptations that the virus has evolved to aid it in escaping immune surveillance, including the establishment of a latent infection in its human host. A latent infection allows the virus to remain in the host without inducing tissue pathology or eliciting an immune response. During the acute infection or reactivation of latent virus, the immune response is significant, which can ultimately result in corneal blindness or fatal sporadic encephalitis. In fact, HSV-1 is one of the leading causes of infectious corneal blindness in the world as a result of chronic episodes of viral reactivation leading to stromal keratitis and scarring. Significant inroads have been made in identifying key immune mediators that control ocular HSV-1 infection and potentially viral reactivation. Likewise, viral mechanisms associated with immune evasion have also been identified and will be discussed. Lastly, novel therapeutic strategies that are currently under development show promise and will be included in this review. Most investigators have taken full advantage of the murine host as a viable working in vivo model of HSV-1 due to the sensitivity and susceptibility to viral infection, ease of manipulation, and a multitude of developed probes to study changes at the cellular and molecular levels. Therefore, comments in this review will primarily be restricted to those observations pertaining to the mouse model and the assumption (however great) that similar events occur in the human condition.

Animals↗
PubMed2005

The trend of susceptibilities to amphotericin B and fluconazole of Candida species from 1999 to 2002 in Taiwan.

BACKGROUND: Candida species have various degrees of susceptibility to common antifungal drugs. The extent of resistance to amphotericin B and fluconazole of Candida glabrata isolates causing candidemia has been reported. Active surveillance may help us to monitor the trend of susceptibility to antifungal drugs and to determine if there is an emerging co-resistance to both drugs of Candida species, specifically, of C. glabrata in Taiwan. METHODS: The susceptibilities to amphotericin B and fluconazole of Candida species collected in 1999 and 2002 of the Taiwan Surveillance of Antimicrobial Resistance of Yeasts (TSARY) were determined by the microdilution method. RESULTS: The antifungal susceptibilities of 342 and 456 isolates collected from 11 hospitals participating in both TSARY 1999 and TSARY 2002, respectively, have been determined. The resistance rate to amphotericin B has increased from 0.3% in the TSARY1999 to 2.2% in the TSARY 2002. In contrast, the resistance rate to fluconazole has decreased from 8.8% to 2.2%. Nevertheless, significantly more C. glabrata isolates were not susceptible to fluconazole in the TSARY 2002 (47.4%) than that in the TSARY 1999 (20.8%). There were 9.8% and 11% of C. glabrata isolates having susceptible-dose dependent and resistant phenotype to fluconazole in the TSARY 1999, verse 45.3% and 2.1% in the TSARY 2002. CONCLUSION: There was an increase of resistance rate to amphotericin B in C. glabrata. On the other hand, although the resistance rate to fluconazole has decreased, almost half of C. glabrata isolates were not susceptible to this drug. Hence, continuous monitoring the emerging of co-resistance to both amphotericin B and fluconazole of Candida species, specifically, of C. glabrata, will be an important early-warning system.

Amphotericin B↗
PubMed2002

Molecular characteristics of Neisseria meningitidis strains isolated in Burkina Faso in 2001.

In the course of an epidemic of meningitis in Burkina Faso in 2001, 27 cerebrospinal fluid samples from patients in 7 districts were forwarded to Norway for isolation and characterization of the causative agents. Neisseria meningitidis was isolated from 13 (48%) samples. The isolates were analysed using serological and genetic methods. Of the 13 strains, 4 were serogroup A, serotype 21:P1.9, sequence type (ST)-5 and belonged to clonal subgroup III, while the remaining 9 strains were serogroup W135, serotype 2a:P1.5,2, ST-11 and belonged to the electrophoretic type-37 complex. PCR analyses revealed meningococcal DNA in 13/14 culture-negative samples. Sequence analysis of the PCR products demonstrated that at least 3 different meningococcal strains were responsible for these 13 cases. Our results show that the W135 strain associated with the 2000 hajj (Muslim pilgrimage) outbreak was a significant cause of disease in Burkina Faso in 2001. Further studies are warranted to determine whether W135 is about to replace serogroup A in sub-Saharan Africa.

Adolescent↗
PubMed2001

A combination of high oxygen atmosphere and equilibrium modified atmosphere packaging to improve the keeping quality of red fruits.

When high-barrier material is used to package fresh whole fruit in high oxygen atmospheres, shelf life is limited due to the accumulation of its produced CO2 and its creation of an anaerobic atmosphere. In this study, the effect of the combination of High Oxygen Atmosphere and Equilibrium Modified Atmosphere Packaging was tested on the shelf life of two non-climacteric red fruits, particularly strawberries and raspberries, at 7 degrees C. Herefor, the following packaging systems are compared: the conventional method of packaging in a macroperforated high-barrier film (air conditions) (1), Equilibrium Modified Atmosphere (EMA, i.e. 3-5% O2 and 5-10% CO2-balance N2) (2) and two novel modified atmospheres: High Oxygen Atmosphere (HOA, i.e. > 70% O2-balance N2) in a high-barrier film (3) and HOA in a EMA film with an adjusted film permeability (4). The shelf life of the fruits was determined by evaluating the evolution of the internal oxygen, carbon dioxide and ethylene concentrations in the packages, the microbial and sensorial quality during the storage, the loss of weight due to transpiration and respiration of the fruit and the loss of marketable fruit due to visual decay or to Botrytis growth. The High Oxygen Atmosphere in the EMA film re-established after five days to an equilibrium of 3% O2 and 5% CO2. The High Oxygen Atmosphere in the high-barrier film remained high during the first five days of storage, but decreased then rapidly to anaerobic conditions, resulting in off-flavours and odours. To avoid an accumulation of ethylene inside the high-barrier package, an ethylene adsorbing monolayer was added. Shelf life of strawberries and raspberries, packed in air conditions, was limited by growth of moulds and yeasts, rather than by sensorial unacceptance. On the other hand, sensorial properties limited the shelf life of the fruits packed under modified atmosphere. Especially High Oxygen Atmosphere improved the microbial quality, due to the inhibiting effect on yeasts and moulds. However, when oxygen was depleted and carbon dioxide had accumulated, sensorial quality (odour, taste andfirmness) was deteriorated. As this is not the case with high O2 in an EMA film, the latter can be assumed as a promising configuration for respiring fresh produce, combining the beneficial effect of high O2 and EMAP.

Bacteria↗
PubMed2004

[An important mission for microbiologists in the new century-cultivation of the unculturable microorganisms].

The present review summarizes the recent advances in cultivating the so-called recalcitrant microorganisms in various natural environments. Firstly, the use of new electron donors and acceptors reveals a list of unique and previously unrecognized physiotypes (Table 1), and the growth supporting reactions include oxidation of phosphite, arsenite, soil humic substances, benzene, Fe (II) and aryl-halides. The novel pure cultures obtained include: Desulfatogenum phosphitoxidans, lithochemotrophic arsenite oxidizing bacteria, Dechloromonas agitata, Dechalosoma suillum, and various Proteobacteria capable of decomposing soil humic substances anaerobically. Secondly, the adaption of nutrient-poor media (down to 1/100 of the traditional concentration), including sea water, soil extract and extension of natural habitats (insect hind-gut, submarine hot vent) are extremely rewarding. For example, cultivation of the ubiquitous SAR11 marine bacterioplankton clade reveals a phylum, small in cell size and genome, but dominating in ocean surface, thus constituting a huge biomass on earth. In addition, the nanosized hyperthermophilic Archaeon from a submarine hot vent, represents an unknown phylum, and because of its symbiotic living with Ignicocous cells, it is named Nanoarchaeum equitans. Its genome size is only 500kb, the smallest for prokaryote. Considering its hyperthermic living environment and small genome, N. equitans may possibly still a primitive form of microbial life and affords an advantageous culture for the study of life origin and evolution on earth. Thirdly, several novel isolation techniques have been developed such as gel microdrop encapsulation and diffusion chamber, both with the advantages of high-throughout operation and simulating to certain extent of the natural environment, thus allowing growth synergy based on cross-feeding to occur. Finally, it should be noted that the cultivation of unculturable microorganisms mentioned in this review does not require expensive and sophisticated equipments and most of the techniques can be adapted in our laboratory at home.

Bacteria↗
PubMed1986

[Use of DNA-DNA hybridization methods for studying the epidemiology and evolutionary similarity of drug resistance determinants].

A procedure for rapid mass assay of clinical strains of microorganisms based on DNA-DNA hybridization was developed. Epidemiology of the gentamicin resistance determinant cloned earlier from clinical strains of the same hospital was studied and its plasmid localization in representatives of various genera of Enterobacteriaceae was confirmed. The studies on dot-hybridization showed that in three antibiotic-producing organisms, i.e. S. fradiae 918 producing neomycin, M. purpurea 1535 producing gentamicin and S. rimosus 65 producing oxytetracycline there were DNA sequences homologous to those of the gentamicin resistance determinant cloned from the clinical isolates.

Biological Evolution↗
PubMed2005

Experimental demonstration of chaos in a microbial food web.

Discovering why natural population densities change over time and vary with location is a central goal of ecological and evolutional disciplines. The recognition that even simple ecological systems can undergo chaotic behaviour has made chaos a topic of considerable interest among theoretical ecologists. However, there is still a lack of experimental evidence that chaotic behaviour occurs in the real world of coexisting populations in multi-species systems. Here we study the dynamics of a defined predator-prey system consisting of a bacterivorous ciliate and two bacterial prey species. The bacterial species preferred by the ciliate was the superior competitor. Experimental conditions were kept constant with continuous cultivation in a one-stage chemostat. We show that the dynamic behaviour of such a two-prey, one-predator system includes chaotic behaviour, as well as stable limit cycles and coexistence at equilibrium. Changes in the population dynamics were triggered by changes in the dilution rates of the chemostat. The observed dynamics were verified by estimating the corresponding Lyapunov exponents. Such a defined microbial food web offers a new possibility for the experimental study of deterministic chaos in real biological systems.

Animals↗
PubMed1996

Primordial emergence of the recombination activating gene 1 (RAG1): sequence of the complete shark gene indicates homology to microbial integrases.

The rearrangement of antibody and T-cell receptor gene segments is indispensable to the vertebrate immune response. All extant jawed vertebrates can rearrange these gene segments. This ability is conferred by the recombination activating genes I and II (RAG I and RAG II). To elucidate their origin and function, the cDNA encoding RAG I from a member of the most ancient class of extant gnathostomes, the Carcharhine sharks, was characterized. Homology domains identified within shark RAG I prompted sequence comparison analyses that suggested similarity of the RAG I and II genes, respectively, to the integrase family genes and integration host factor genes of the bacterial site-specific recombination system. Thus, the apparent explosive evolution (or "big bang") of the ancestral immune system may have been initiated by a transfer of microbial site-specific recombinases.

Amino Acid Sequence↗
PubMed2000

Adaptive evolution of relish, a Drosophila NF-kappaB/IkappaB protein.

NF-kappaB and IkappaB proteins have central roles in regulation of inflammation and innate immunity in mammals. Homologues of these proteins also play an important role in regulation of the Drosophila immune response. Here we present a molecular population genetic analysis of Relish, a Drosophila NF-kappaB/IkappaB protein, in Drosophila simulans and D. melanogaster. We find strong evidence for adaptive protein evolution in D. simulans, but not in D. melanogaster. The adaptive evolution appears to be restricted to the IkappaB domain. A possible explanation for these results is that Relish is a site of evolutionary conflict between flies and their microbial pathogens.

Adaptation, Biological↗
PubMed2003

Molecular strategies in biological evolution of antimicrobial peptides.

Gene-encoded antimicrobial peptides that protect the skin of hylid and ranin frogs against noxious microorganisms are processed from a unique family of precursor polypeptides with a unique pattern of conserved and variable regions opposite to that of conventional secreted peptides. Precursors belonging to this family, designated the preprodermaseptin, have a common N-terminal preproregion that is remarkably well conserved both within and between species, but a hypervariable C-terminal domain corresponding to antimicrobial peptides with very different lengths, sequences, charges and antimicrobial spectra. Each frog species has its own distinct panoply of 10-20 antimicrobial peptides so that the 5000 species of ranids and hylids may produce approximately 100,000 different peptide antibiotics. The strategy that these frogs have evolved to generate this enormous array of peptides includes repeated duplications of a 150 million years old ancestral gene, focal hypermutation of the antimicrobial peptide domain maybe involving a mutagenic DNA polymerase similar to Escherichia coli Pol V, and subsequent actions of positive (diversifying) selection. The hyperdivergence of skin antimicrobial peptides can be viewed as the successful evolution of a multi-drug defense system that provides frogs with maximum protection against rapidly changing microbial biota and minimizes the chance of microorganisms developing resistance to individual peptides. The impressive variations in the expression of frog skin antimicrobial peptides may be exploited for discovering new molecules and structural motifs targeting specific microorganisms for which the therapeutic armamentarium is scarce.

Amino Acid Sequence↗