Spatial and temporal control of lysis by the lambda holin.

The infection cycle of phage λ terminates in lysis mediated by three types of lysis proteins, each disrupting a layer in the bacterial envelope: the S105 holin, the R endolysin, and the Rz/Rz1 spanin complex targeting the inner membrane, cell wall or peptidoglycan, and the outer membrane, respectively. Video microscopy has shown that in most infections, lysis occurs as a sudden, explosive event at a cell pole, such that the initial product is a less refractile ghost that retains rod-shaped morphology. Here, we investigate the molecular basis of polar lysis using time-lapse fluorescence microscopy. The results indicate that the holin determines the morphology of lysis by suddenly forming two-dimensional rafts at the poles about 100 s prior to lysis. Given the physiological and biochemical similarities between the lambda holin and other class I holins, dynamic redistribution and sudden concentration may be common features of holins, probably reflecting the fitness advantage of all-or-nothing lysis regulation.IMPORTANCEIn this study, we use fluorescent video microscopy to track -green fluorescent protein (GFP)-labeled holin in the minutes prior to phage lysis. Our work contextualizes prior genetic and biochemical data, showing when hole formation starts and where holin oligomers form in relation to the site of lytic rupture. Furthermore, prior work showed that the morphology of lambda-infected cells is characterized by an explosive event starting at the cell pole; however, the basis for this was not clear. This study shows that holin most often oligomerizes at cell poles and that the site of the oligomerization is spatially correlated with the site of lytic blowout. Therefore, the holin is the key contributor to polar lysis morphology for phage lambda.

Mechanisms of pathogenicity for the emerging fungus Candida auris.

Candida auris recently emerged as an urgent public health threat, causing outbreaks of invasive infections in healthcare settings throughout the world. This fungal pathogen persists on the skin of patients and on abiotic surfaces despite antiseptic and decolonization attempts. The heightened capacity for skin colonization and environmental persistence promotes rapid nosocomial spread. Following skin colonization, C. auris can gain entrance to the bloodstream and deeper tissues, often through a wound or an inserted medical device, such as a catheter. C. auris possesses a variety of virulence traits, including the capacity for biofilm formation, production of adhesins and proteases, and evasion of innate immune responses. In this review, we highlight the interactions of C. auris with the host, emphasizing the intersection of laboratory studies and clinical observations.

Phage-encoded cationic antimicrobial peptide required for lysis.

Most phages of Gram-negative hosts encode spanins for disruption of the outer membrane, the last step in host lysis. However, bioinformatic analysis indicates that ∼15% of these phages lack a spanin gene, suggesting they have an alternate way of disrupting the OM. Here, we show that the T7-like coliphage phiKT causes the explosive cell lysis associated with spanin activity despite not encoding spanins. A putative lysis cassette cloned from the phiKT late gene region includes the hypothetical novel gene 28 located between the holin and endolysin genes and supports inducible lysis in E. coli K-12. Moreover, induction of an isogenic construct lacking gene 28 resulted in divalent cation-stabilized spherical cells rather than lysis, implicating gp28 in OM disruption. Additionally, gp28 was shown to complement the lysis defect of a spanin-null λ lysogen. Gene 28 encodes a 56-amino acid cationic protein with predicted amphipathic helical structure and is membrane-associated after lysis. Urea and KCl washes did not release gp28 from the particulate, suggesting a strong hydrophobic membrane interaction. Fluorescence microscopy supports membrane localization of the gp28 protein prior to lysis. Gp28 is similar in size, charge, predicted fold, and membrane association to the human cathelicidin antimicrobial peptide LL-37. Synthesized gp28 behaved similar to LL-37 in standard assays mixing peptide and cells to measure bactericidal and inhibitory effects. Taken together, these results indicate that phiKT gp28 is a phage-encoded cationic antimicrobial peptide that disrupts bacterial outer membranes during host lysis and thus establishes a new class of phage lysis proteins, the disruptins. Significance We provide evidence that phiKT produces an antimicrobial peptide for outer membrane disruption during lysis. This protein, designated as a disruptin, is a new paradigm for phage lysis and has no similarities to other known lysis genes. Although many mechanisms have been proposed for the function of antimicrobial peptides, there is no consensus on the molecular basis of membrane disruption. Additionally, there is no established genetic system to support such studies. Therefore, the phiKT disruptin may represent the first genetically tractable antimicrobial peptide, facilitating mechanistic analyses.

Complete Genome Sequence of Escherichia coli Phage Paul.

Escherichia coli is both a commensal and a pathogen in humans and other animals. Here, we describe the isolation of E. coli strain 4s bacteriophage Paul. The complete 79,429-bp genome was annotated and demonstrates similarity with phieco32viruses, as does its prolate podophage morphology.

Suppressor Analysis of the Fusogenic Lambda Spanins.

The final step of lysis in phage λ infections of Escherichia coli is mediated by the spanins Rz and Rz1. These proteins form a complex that bridges the cell envelope and that has been proposed to cause fusion of the inner and outer membranes. Accordingly, mutations that block spanin function are found within coiled-coil domains and the proline-rich region, motifs essential in other fusion systems. To gain insight into spanin function, pseudorevertant alleles that restored plaque formation for lysis-defective mutants of Rz and Rz1 were selected. Most second-site suppressors clustered within a coiled-coil domain of Rz near the outer leaflet of the cytoplasmic membrane and were not allele specific. Suppressors largely encoded polar insertions within the hydrophobic core of the coiled-coil interface. Such suppressor changes resulted in decreased proteolytic stability of the Rz double mutants in vivo Unlike the wild type, in which lysis occurs while the cells retain a rod shape, revertant alleles with second-site suppressor mutations supported lysis events that were preceded by spherical cell formation. This suggests that destabilization of the membrane-proximal coiled coil restores function for defective spanin alleles by increasing the conformational freedom of the complex at the cost of its normal, all-or-nothing functionality.IMPORTANCECaudovirales encode cell envelope-spanning proteins called spanins, which are thought to fuse the inner and outer membranes during phage lysis. Recent genetic analysis identified the functional domains of the lambda spanins, which are similar to class I viral fusion proteins. While the pre- and postfusion structures of model fusion systems have been well characterized, the intermediate structure(s) formed during the fusion reaction remains elusive. Genetic analysis would be expected to identify functional connections between intermediates. Since most membrane fusion systems are not genetically tractable, only few such investigations have been reported. Here, we report a suppressor analysis of lambda spanin function. To our knowledge this is the first suppression analysis of a class I-like complex and also the first such analysis of a prokaryote membrane fusion system.

Genetic Analysis of the Lambda Spanins Rz and Rz1: Identification of Functional Domains.

Coliphage lambda proteins Rz and Rz1 are the inner membrane and outer membrane subunits of the spanin complex-a heterotetramer that bridges the periplasm and is essential for the disruption of the outer membrane during phage lysis. Recent evidence suggests the spanin complex functions by fusing the inner and outer membrane. Here, we use a genetics approach to investigate and characterize determinants of spanin function. Because Rz1 is entirely embedded in the +1 reading frame of Rz, the genes were disembedded before using random mutagenesis to construct a library of lysis-defective alleles for both genes. Surprisingly, most of the lysis-defective missense mutants exhibited normal accumulation or localization in vivo, and also were found to be normal for complex formation in vitro Analysis of the distribution and nature of single missense mutations revealed subdomains that resemble key motifs in established membrane-fusion systems, i.e., two coiled-coil domains in Rz, a proline-rich region of Rz1, and flexible linkers in both proteins. When coding sequences are aligned respective to the embedded genetic architecture of Rz1 within Rz, genetically silent domains of Rz1 correspond to mutationally sensitive domains in Rz, and vice versa, suggesting that the modular structure of the two subunits facilitated the evolutionary compression that resulted in the unique embedded gene architecture.

Is an average of routine postdialysis blood pressure a good indicator of blood pressure control and cardiovascular risk?

INTRODUCTION: Hypertension is associated with left ventricular hypertrophy (LVH), a predictor of cardiovascular mortality in haemodialysis (HD) patients. The National Kidney Foundation Kidney Disease Outcomes Quality Initiative (NKF-KDOQI) blood pressure (BP) targets are pre-HD <140/90 mm Hg, post-HD <130/80 mm Hg. This study aims to assess 3-month mean in-unit BP, pre- and post-HD, for correlations with left ventricular mass index (LVMI), a measure of long-term BP control. METHODS: Of 648 HD patients, including those on HD >6 months, 262 had echocardiograms. Those with significant coronary artery disease, reduced ejection fraction or valvular disease were excluded, as were those without appropriate echocardiogram, leaving 100 patients. Data on BP and confounding factors for LVH were collected covering 3 months prior to echocardiogram. RESULTS: Mean BP pre-HD was 147/77 ± 19/13 mm Hg, and post-HD, 133/71 ± 20/11 mm Hg; <50% of patients achieved NKF targets. Mean LVMI was 203.7 ± 74 g/m(2); 88% of patients had LVH. On univariate analysis, mean pre- and post-HD systolic BP, mean arterial blood pressure (MAP) and post-HD diastolic and pulse pressure correlated with LVMI. On stepwise multiple regression analysis only post-HD MAP correlated with LVMI (p=0.000047, r=0.395). CONCLUSIONS: We conclude that long-term averages of in-unit post-HD BP measurements are useful in assessing BP control and cardiovascular risk, especially in the absence of routine ambulatory or home BP monitoring.

Spatial analysis of plague in California: niche modeling predictions of the current distribution and potential response to climate change.

BACKGROUND: Plague, caused by the bacterium Yersinia pestis, is a public and wildlife health concern in California and the western United States. This study explores the spatial characteristics of positive plague samples in California and tests Maxent, a machine-learning method that can be used to develop niche-based models from presence-only data, for mapping the potential distribution of plague foci. Maxent models were constructed using geocoded seroprevalence data from surveillance of California ground squirrels (Spermophilus beecheyi) as case points and Worldclim bioclimatic data as predictor variables, and compared and validated using area under the receiver operating curve (AUC) statistics. Additionally, model results were compared to locations of positive and negative coyote (Canis latrans) samples, in order to determine the correlation between Maxent model predictions and areas of plague risk as determined via wild carnivore surveillance. RESULTS: Models of plague activity in California ground squirrels, based on recent climate conditions, accurately identified case locations (AUC of 0.913 to 0.948) and were significantly correlated with coyote samples. The final models were used to identify potential plague risk areas based on an ensemble of six future climate scenarios. These models suggest that by 2050, climate conditions may reduce plague risk in the southern parts of California and increase risk along the northern coast and Sierras. CONCLUSION: Because different modeling approaches can yield substantially different results, care should be taken when interpreting future model predictions. Nonetheless, niche modeling can be a useful tool for exploring and mapping the potential response of plague activity to climate change. The final models in this study were used to identify potential plague risk areas based on an ensemble of six future climate scenarios, which can help public managers decide where to allocate surveillance resources. In addition, Maxent model results were significantly correlated with coyote samples, indicating that carnivore surveillance programs will continue to be important for tracking the response of plague to future climate conditions.

Impact of changing water levels and weather on Oncomelania hupensis hupensis populations, the snail host of Schistosoma japonicum, downstream of the Three Gorges Dam.

Increasing evidence indicates that dams impact riverine ecosystems and human diseases. Poyang Lake, one of the largest schistosomiasis endemic environments in China, will change due to the construction of the Yangtze River Three Gorges Dam. We assess changes in Oncomelania hupensis hupensis, the snail host for Schistosoma japonicum, in response to changing water levels and weather from 1998 to 2002. In the 5 years following the major flooding of Poyang Lake in 1998, seasonal water levels have gradually decreased, concomitant with decreases in mean and variance of fall snail densities. Nonlinear relationships suggest that the highest spring density is associated with current, 2-, and 3-month prior temperatures of 18 degrees, 9.1 degrees, and 5.8 degrees C, while the highest fall density is associated with 2- and 3-month prior water levels of 17 and 18 m, respectively. This suggests that lower, more stable water levels downstream of the dam may result in a reduction in mean fall densities and their variance. However, additional data are needed to determine whether snail populations that are typically destroyed by seasonal floods may live longer in more stable environments created by the dam.

Spatial distribution of traffic induced noise exposures in a US city: an analytic tool for assessing the health impacts of urban planning decisions.

BACKGROUND: Vehicle traffic is the major source of noise in urban environments, which in turn has multiple impacts on health. In this paper we investigate the spatial distribution of community noise exposures and annoyance. Traffic data from the City of San Francisco were used to model noise exposure by neighborhood and road type. Remote sensing data were used in the model to estimate neighborhood-specific percentages of cars, trucks, and buses on arterial versus non-arterial streets. The model was validated on 235 streets. Finally, an exposure-response relationship was used to predict the prevalence of high annoyance for different neighborhoods. RESULTS: Urban noise was found to increase 6.7 dB (p < 0.001) with 10-fold increased street traffic, with important contributors to noise being bus and heavy truck traffic. Living along arterial streets also increased risk of annoyance by 40%. The relative risk of annoyance in one of the City's fastest growing neighborhoods, the South of Market Area, was found to be 2.1 times that of lowest noise neighborhood. However, higher densities of exposed individuals were found in Chinatown and Downtown/Civic Center. Overall, we estimated that 17% of the city's population was at risk of high annoyance from traffic noise. CONCLUSION: The risk of annoyance from urban noise is large, and varies considerably between neighborhoods. Such risk should be considered in urban areas undergoing rapid growth. We present a relatively simple GIS-based noise model that may be used for routinely evaluating the health impacts of environmental noise.

An easy method for interpreting the results of arterial blood gas analysis.

Interpretation of acid-base disturbances is an essential skill for critical care nurses. Using the H model makes this process easy. When students and novice critical care nurses feel competent with certain skills, their confidence levels are greatly enhanced. One of us (K.M.K.) has been using the H model for many years to teach students how to interpret the results of arterial blood gas analysis. The students are often amazed at how easy and fun the model makes learning a subject many perceive as complex.

Nurses pray: use of prayer and spirituality as a complementary therapy in the intensive care setting.

The power that prayer and spirituality exerts on healing cannot be underestimated. Body, mind, and spirit are connected to each other. Although patients in hospitals may have the best medical and nursing care available, many seek alternative or complementary therapies. One adjunctive therapy that has grown in popularity recently is the incorporation of prayer and spirituality into the traditional approaches used with acute and critically ill patients. Spirituality is returning to healthcare because many patients believe in it and seek it as part of their treatment. Although spirituality is only one of the many types of alternative and complementary therapies available to patients, it can be powerful approach to their care. This article explores the use of spirituality with a special focus on prayer.

Autoerotic asphyxia in adolescents.

The death of a child or young adult is always tragic, regardless of the cause. Cases of autoerotic asphyxia are often labeled as suicide, or are underreported because of embarrassment of relatives or misidentification of the initial clinical manifestations. It may be that autoerotic asphyxial death is far more common than realized. Many emergency nurses and physicians lack adequate knowledge about this phenomenon to make an accurate diagnosis. Family members are often reluctant or unwilling to provide enough data surrounding the circumstances in which the patient was found, and the cause of death is mislabeled as suicide. Autoerotic asphyxia is frequently labeled as a sexual aberrancy and an act that society would rather not acknowledge. But there are a number of implications for emergency nurses, such as prevention and sensitive support of family in the emergency department, that demand our attention.