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PubMed2026

Gabija restricts phage circularization and DNA replication.

Anti-bacteriophage systems such as restriction-modification and CRISPR-Cas have DNA substrate specificity mechanisms that enable the identification of invaders. How Gabija, a highly prevalent nuclease-helicase antiphage system, limits phage replication while executing self- vs. non-self-discrimination remains unknown. Here, we show that phage-encoded DNA end-binding proteins that antagonize host RecBCD sensitize phages to Gabija. When targeting a temperate lambda-like phage in Pseudomonas aeruginosa, Gabija prevents phage genome circularization and subsequent replication. DNA end-binding complexes, including a phage exonuclease and a single-stranded DNA (ssDNA)-annealing protein or GamMu dimers that prevent loading of the host repair complex RecBCD, are necessary and sufficient to license phage and plasmid sensitivity to Gabija. Mutant escape phages lacking these DNA end-binding proteins become protected from Gabija by RecBCD translocation activities. RecBCD activity on the bacterial genome, presumably whenever it is linearized, also prevents Gabija from targeting self-DNA. Therefore, we propose that Gabija antagonizes the circularization and replication of linear DNA devoid of RecBCD as a mechanism to identify and antagonize foreign invaders.

DNA Replication↗
PubMed2026

Extensive hidden prophage diversity in Enterobacter species reveals host specificity and local distribution.

Bacteriophages are key drivers of bacterial evolution, particularly through their integration as prophages within host genomes. However, the diversity and host specificity of prophages in relevant pathogens such as Enterobacter species remain poorly characterized. In this study, we revealed the diversity of prophages, mapped their distribution and explored their relationships with their bacterial hosts. We analysed 3,661 prophage sequences identified from the genomes of 20 different Enterobacter species. This analysis uncovered an extensive hidden diversity, comprising 1,617 phage genera and 2,423 phage species - nearly 80% of which were singletons - highlighting an exceptionally rich prophage landscape. We found substantial variation in prophage species richness across host species and isolation sources, with Enterobacter kobei and environmental isolates exhibiting the highest richness. Prophage populations showed strong host specificity and limited cross-species transmission. Moreover, prophages exhibited geographic structuring and significant congruence between host and prophage phylogenies, as well as with the ecological lifestyles of their bacterial hosts. Although we found phages of the same species infecting different host species, these events were infrequent. Finally, bacterial genomes encoded diverse defence systems, mainly PDC-S07, RM type I-II and gabija, whereas only 8.9% of prophages encoded anti-defence systems, mostly anti-CBASS and anti-RM. Overall, this study provides new insights into the diversity of Enterobacter prophages and underscores their ecological and clinical relevance in shaping host adaptation and phage-host dynamics.

Prophages↗
PubMed2006

Diagnostic accuracy of 64-slice multislice computed tomography in the noninvasive evaluation of significant coronary artery disease.

The purpose of the present study was to determine the diagnostic accuracy of current 64-slice multislice computed tomography (MSCT) in the detection of significant coronary artery disease, using conventional coronary angiography as the gold standard. In 61 patients scheduled for conventional coronary angiography, 64-slice MSCT was performed and evaluated for the presence of significant (>or=50% luminal narrowing) stenoses. One patient had to be excluded because of a heart rate>90 beats/min during data acquisition. In the remaining 60 patients (46 men, 14 women; average age 60+/-11 years), 854 segments were available for evaluation. Of these segments 842 (99%) were of sufficient image quality. Conventional coronary angiography identified 73 lesions, of which 62 were detected by MSCT. The corresponding sensitivity and specificity were 85% and 97%, respectively. On a patient-per-patient analysis, sensitivity, specificity, and positive and negative predictive values were 94%, 97%, 97%, and 93%, respectively. In conclusion, the present study confirms that 64-slice MSCT enables the accurate and noninvasive evaluation of significant coronary artery stenoses.

Adolescent↗
PubMed2006

Prognostic value of multislice computed tomography coronary angiography in patients with known or suspected coronary artery disease.

OBJECTIVES: This study sought to determine the prognostic value of multislice computed tomography (MSCT) coronary angiography in patients with known or suspected coronary artery disease (CAD). BACKGROUND: It is expected that MSCT will be used increasingly as an alternative imaging modality in the diagnosis of patients with suspected CAD. Data on the prognostic value of MSCT, however, are currently not available. METHODS: A total of 100 patients (73 men, age 59 +/- 12 years) who were referred for further cardiac evaluation due to suspicion of significant CAD underwent additional MSCT coronary angiography to evaluate the presence and severity of CAD. Patients were followed up for the occurrence of: 1) cardiac death, 2) nonfatal myocardial infarction, 3) unstable angina requiring hospitalization, and 4) revascularization. RESULTS: Coronary plaques were detected in 80 (80%) patients. During a mean follow-up of 16 months, 33 events occurred in 26 patients. In patients with normal coronary arteries on MSCT, the first-year event rate was 0% versus 30% in patients with any evidence of CAD on MSCT. The observed event rate was highest in the presence of obstructive lesions (63%) and when obstructive lesions were located in the left main (LM)/left anterior descending (LAD) coronary arteries (77%). Nonetheless, an elevated event rate was also observed in patients with nonobstructive CAD (8%). In multivariate analysis, significant predictors of events were the presence of CAD, obstructive CAD, obstructive CAD in LM/LAD, number of segments with plaques, number of segments with obstructive plaques, and number of segments with mixed plaques. CONCLUSIONS: Multislice computed tomography coronary angiography provides independent prognostic information over baseline clinical risk factors in patients with known and suspected CAD. An excellent prognosis was noted in patients with a normal MSCT.

Aged↗
PubMed2005

The Holin protein of bacteriophage PRD1 forms a pore for small-molecule and endolysin translocation.

PRD1 is a bacteriophage with an icosahedral outer protein layer surrounding the viral membrane, which encloses the linear double-stranded DNA genome. PRD1 infects gram-negative cells harboring a conjugative IncP plasmid. Here we studied the lytic functions of PRD1. Using infected cells and plasmid-borne lysis genes, we demonstrated that a two-component lysis system (holin-endolysin) operates to release progeny phage particles from the host cell. Monitoring of ion fluxes and the ATP content of the infected cells allowed us to build a model of the sequence of lysis-related physiological changes. A decrease in the intracellular level of ATP is the earliest indicator of cell lysis, followed by the leakage of K+ from the cytosol approximately 20 min prior to the decrease in culture turbidity. However, the K+ efflux does not immediately lead to the depolarization of the cytoplasmic membrane or leakage of the intracellular ATP. These effects are observed only approximately 5 to 10 min prior to cell lysis. Similar results were obtained using cells expressing the holin and endolysin genes from plasmids.

Bacteriolysis↗
PubMed2007

Efficient DNA packaging of bacteriophage PRD1 requires the unique vertex protein P6.

The assembly of bacteriophage PRD1 proceeds via formation of empty procapsids containing an internal lipid membrane, into which the linear double-stranded DNA genome is subsequently packaged. The packaging ATPase P9 and other putative packaging proteins have been shown to be located at a unique vertex of the PRD1 capsid. Here, we describe the isolation and characterization of a suppressor-sensitive PRD1 mutant deficient in the unique vertex protein P6. Protein P6 was found to be an essential part of the PRD1 packaging machinery; its absence leads to greatly reduced packaging efficiency. Lack of P6 was not found to affect particle assembly, because in the P6-deficient mutant infection, wild-type (wt) amounts of particles were produced, although most were empty. P6 was determined not to be a specificity factor, as the few filled particles seen in the P6-deficient infection contained only PRD1-specific DNA. The presence of P6 was not necessary for retention of DNA in the capsid once packaging had occurred, and P6-deficient DNA-containing particles were found to be stable and infectious, albeit not as infectious as wt PRD1 virions. A packaging model for bacteriophage PRD1, based on previous results and those obtained in this study, is presented.

Bacteriophage PRD1↗
PubMed2006

Advances in the noninvasive evaluation of coronary artery disease with multislice computed tomography.

Current noninvasive detection of coronary artery disease (CAD) is based on the demonstration of ischemia using stress-rest imaging, which is an indirect way of identifying CAD by demonstration of the hemodynamic consequences, rather than direct visualization of the obstructive lesions in the coronary arteries. Multislice computed tomography (MSCT) has recently emerged as an extremely rapidly developing noninvasive imaging modality, which allows anatomical imaging of the coronary arteries or noninvasive coronary angiography. In addition, total plaque burden, plaque morphology and (to some extent) plaque constitution can be assessed by MSCT. The technique also provides information on resting left ventricular systolic function, and possibly resting perfusion. Ideally, stress function and perfusion should also be evaluated, since this would allow detection of ischemia and would complete the picture of CAD. However, this is not routinely performed, since sequential acquisitions are associated with high radiation doses and thus pose a limitation for cardiovascular applications of MSCT. It is anticipated that, with a reduction in radiation, MSCT may become an important player in the diagnostic and prognostic work-up of patients with known or suspected CAD.

Coronary Angiography↗