Lytic bacteriophages induce the secretion of antiviral and proinflammatory cytokines from human respiratory epithelial cells.

Phage therapy is a therapeutic approach to treat multidrug-resistant (MDR) infections that employs lytic bacteriophages (phages) to eliminate bacteria. Despite the abundant evidence for its success as an antimicrobial in Eastern Europe, there is scarce data regarding its effects on the human host. Here, we aimed to understand how lytic phages interact with cells of the airway epithelium, the tissue site that is colonized by bacterial biofilms in numerous chronic respiratory disorders. Using a panel of Pseudomonas aeruginosa phages and human airway epithelial cells (AECs) derived from a person with cystic fibrosis (CF), we determined that interactions between phages and epithelial cells depend on specific phage properties as well as physiochemical features of the microenvironment. Although poor at internalizing phages, the airway epithelium responds to phage exposure by changing its transcriptional profile and secreting antiviral and proinflammatory cytokines that correlate with specific phage families. Overall, our findings indicate that mammalian responses to phages are heterogenous and could potentially alter the way that respiratory local defenses aid in bacterial clearance during phage therapy. Thus, besides phage receptor specificity in a particular bacterial isolate, the criteria to select lytic phages for therapy should be expanded to include mammalian cell responses.

Gender-responsiveness in corrections: Estimating female inmate misconduct risk using the Personality Assessment Inventory (PAI).

Proper inmate assessment is critical to correctional management and institutional security. While many instruments have been developed to assist with this process, most of these tools have not been validated using samples of female inmates although distinct gender differences have been identified in the inmate population in terms of adaptation and misconduct. The Personality Assessment Inventory (PAI) is a multiscale measure of psychopathology that is being increasingly utilized in the correctional setting to assist with the inmate classification process. The current study contributes to the dearth of literature surrounding gender-responsive inmate classification by utilizing a sample of 2,000 female inmates to examine the incremental and predictive validity of the PAI in association with general and assaultive disciplinary infractions. Findings from this study reveal that the PAI scales presenting the strongest relationship to general and assaultive disciplinary infractions among this female sample included Aggression (AGG), Antisocial Features (ANT), Paranoia (PAR), and the Violence Potential Index (VPI). Moreover, findings derived from this study suggest that certain PAI measures, specifically ARD-T, DRU, and more general substance abuse and mental health indicators may be useful in gender-responsive assessments during the female inmate classification process.

Homotrimeric macrophage migration inhibitory factor (MIF) drives inflammatory responses in the corneal epithelium by promoting caveolin-rich platform assembly in response to infection.

Acute inflammation that arises during Pseudomonas aeruginosa-induced ocular infection can trigger tissue damage resulting in long term impairment of visual function, suggesting that the appropriate treatment strategy should include the use of anti-inflammatory agents in addition to antibiotics. We recently identified a potential target for modulation during ocular infection, macrophage migration inhibitory factor (MIF). MIF deficiency protected mice from inflammatory-mediated corneal damage resulting from acute bacterial keratitis. To gain a better understanding of the molecular mechanisms of MIF activity, we analyzed the oligomeric states and functional properties of MIF during infection. We found that in human primary corneal cells infected with P. aeruginosa, MIF is primarily in a homotrimeric state. Homotrimeric MIF levels correlated with the severity of infection in the corneas of infected mice, suggesting that the MIF homotrimers were the functionally active form of MIF. During infection, human primary corneal cells released more IL-8 when treated with recombinant, locked MIF trimers than when treated with lower MIF oligomers. MIF promoted P. aeruginosa-induced IL-8 responses via the formation of caveolin-1-rich "signaling hubs" in the corneal cells that led to elevated MAPK p42/p44 activation and sustained inflammatory signaling. These findings suggest that inhibiting homotrimerization of MIF or the functional activities of MIF homotrimers could have therapeutic benefits during ocular inflammation.

Prolonged activity of a recombinant factor VIII-Fc fusion protein in hemophilia A mice and dogs.

Despite proven benefits, prophylactic treatment for hemophilia A is hampered by the short half-life of factor VIII. A recombinant factor VIII-Fc fusion protein (rFVIIIFc) was constructed to determine the potential for reduced frequency of dosing. rFVIIIFc has an ∼ 2-fold longer half-life than rFVIII in hemophilia A (HemA) mice and dogs. The extension of rFVIIIFc half-life requires interaction of Fc with the neonatal Fc receptor (FcRn). In FcRn knockout mice, the extension of rFVIIIFc half-life is abrogated, and is restored in human FcRn transgenic mice. The Fc fusion has no impact on FVIII-specific activity. rFVIIIFc has comparable acute efficacy as rFVIII in treating tail clip injury in HemA mice, and fully corrects whole blood clotting time (WBCT) in HemA dogs immediately after dosing. Furthermore, consistent with prolonged half-life, rFVIIIFc shows 2-fold longer prophylactic efficacy in protecting HemA mice from tail vein transection bleeding induced 24-48 hours after dosing. In HemA dogs, rFVIIIFc also sustains partial correction of WBCT 1.5- to 2-fold longer than rFVIII. rFVIIIFc was well tolerated in both species. Thus, the rescue of FVIII by Fc fusion to provide prolonged protection presents a novel pathway for FVIII catabolism, and warrants further investigation.

Fast-evolving cofactors regulate the role of HEATR5 complexes in intra-Golgi trafficking.

The highly conserved HEATR5 proteins are best known for their roles in membrane traffic mediated by the adaptor protein complex-1 (AP1). HEATR5 proteins rely on fast-evolving cofactors to bind to AP1. However, how HEATR5 proteins interact with these cofactors is unknown. Here, we report that the budding yeast HEATR5 protein, Laa1, functions in two biochemically distinct complexes. These complexes are defined by a pair of mutually exclusive Laa1-binding proteins, Laa2 and the previously uncharacterized Lft1/Yml037c. Despite limited sequence similarity, biochemical analysis and structure predictions indicate that Lft1 and Laa2 bind Laa1 via structurally similar mechanisms. Both Laa1 complexes function in intra-Golgi recycling. However, only the Laa2-Laa1 complex binds to AP1 and contributes to its localization. Finally, structure predictions indicate that human HEATR5 proteins bind to a pair of fast-evolving interacting partners via a mechanism similar to that observed in yeast. These results reveal mechanistic insight into how HEATR5 proteins bind their cofactors and indicate that Laa1 performs functions besides recruiting AP1.

Lytic bacteriophages interact with respiratory epithelial cells and induce the secretion of antiviral and proinflammatory cytokines.

Phage therapy is a therapeutic approach to treat multidrug resistant infections that employs lytic bacteriophages (phages) to eliminate bacteria. Despite the abundant evidence for its success as an antimicrobial in Eastern Europe, there is scarce data regarding its effects on the human host. Here, we aimed to understand how lytic phages interact with cells of the airway epithelium, the tissue site that is colonized by bacterial biofilms in numerous chronic respiratory disorders. We determined that interactions between phages and epithelial cells depend on specific phage properties as well as physiochemical features of the microenvironment. Although poor at internalizing phages, the airway epithelium responds to phage exposure by changing its transcriptional profile and secreting antiviral and proinflammatory cytokines that correlate with specific phage families. Overall, our findings indicate that mammalian responses to phages are heterogenous and could potentially alter the way that respiratory local defenses aid in bacterial clearance during phage therapy. Thus, besides phage receptor specificity in a particular bacterial isolate, the criteria to select lytic phages for therapy should be expanded to include mammalian cell responses.

Probability of criminal acts of violence: a test of jury predictive accuracy.

The ability of capital juries to accurately predict future prison violence at the sentencing phase of aggravated murder trials was examined through retrospective review of the disciplinary records of 115 male inmates sentenced to either life (n = 65) or death (n = 50) in Oregon from 1985 through 2008, with a mean post-conviction time at risk of 15.3 years. Violent prison behavior was completely unrelated to predictions made by capital jurors, with bidirectional accuracy simply reflecting the base rate of assaultive misconduct in the group. Rejection of the special issue predicting future violence enjoyed 90% accuracy. Conversely, predictions that future violence was probable had 90% error rates. More than 90% of the assaultive rule violations committed by these offenders resulted in no harm or only minor injuries.

Two functionally distinct HEATR5 protein complexes are defined by fast-evolving co-factors in yeast.

The highly conserved HEATR5 proteins are best known for their roles in membrane traffic mediated by the adaptor protein complex-1 (AP1). HEATR5 proteins rely on fast-evolving co-factors to bind to AP1. However, how HEATR5 proteins interact with these co-factors is unknown. Here, we report that the budding yeast HEATR5 protein, Laa1, functions in two biochemically distinct complexes. These complexes are defined by a pair of mutually exclusive Laa1-binding proteins, Laa2 and the previously uncharacterized Lft1/Yml037c. Despite limited sequence similarity, biochemical analysis and structure predictions indicate that Lft1 and Laa2 bind Laa1 via structurally similar mechanisms. Both Laa1 complexes function in intra-Golgi recycling. However, only the Laa2-Laa1 complex binds to AP1 and contributes to its localization. Finally, structure predictions indicate that human HEATR5 proteins bind to a pair of fast-evolving interacting partners via a mechanism similar to that observed in yeast. These results reveal mechanistic insight into how HEATR5 proteins bind their co-factors and indicate that Laa1 performs functions besides recruiting AP1.

Prolonged activity of factor IX as a monomeric Fc fusion protein.

Treatment of hemophilia B requires frequent infusions of factor IX (FIX) to prophylax against bleeding episodes. Hemophilia B management would benefit from a FIX protein with an extended half-life. A recombinant fusion protein (rFIXFc) containing a single FIX molecule attached to the Fc region of immunoglobulin G was administered intravenously and found to have an extended half-life, compared with recombinant FIX (rFIX) in normal mice, rats, monkeys, and FIX-deficient mice and dogs. Recombinant FIXFc protein concentration was determined in all species, and rFIXFc activity was measured in FIX-deficient animals. The half-life of rFIXFc was approximately 3- to 4-fold longer than that of rFIX in all species. In contrast, in mice in which the neonatal Fc receptor (FcRn) was deleted, the half-life of rFIXFc was similar to rFIX, confirming the increased circulatory time was due to protection of the rFIXFc via the Fc/FcRn interaction. Whole blood clotting time in FIX-deficient mice was corrected through 144 hours for rFIXFc, compared with 72 hours for rFIX; similar results were observed in FIX-deficient dogs. Taken together, these studies show the enhanced pharmacodynamic and pharmacokinetic properties of the rFIXFc fusion protein and provide the basis for evaluating rFIXFc in patients with hemophilia B.

Community violence to prison assault: a test of the behavioral continuity hypothesis.

This large-scale study (N = 23,277) investigated the relationship between criminal history in the community and serious or assaultive prison misconduct, while controlling for the effects of inmate characteristics, general criminality, and custody level. Community violence variables included the rate of prior violent crime arrests and the types of prior violent crime, as well as a range of specific violent crimes of conviction. Behavioral continuity from community to prison was neither simple nor intuitively discernible, depending on the type, recency, and pattern of community criminality. Application of logistic regression models revealed that the omnibus measure of the rate of prior violent arrests was not related to either serious or assaultive prison misconduct. Prior arrests for assault and current convictions for robbery and/or assault, but not prior or current homicides, were associated with an increased risk for prison violence. Current conviction for a sexual assault had the strongest inverse relationship to prison violence, while prior arrests for sexual assault showed no relationship to prison violence. A more "nuanced" approach in assigning risk ratings based on prior criminal history and seriousness of offense is recommended.

Prison Homicide: An Extension of Violent Criminal Careers?

This study investigated prison homicide perpetrators through the lens of the career criminal perspective. Prison homicide, while a rare event, has critical implications for the prison environment. Despite its importance as a form of institutional violence that must be addressed, only four studies in the past five decades have explored the characteristics of homicide perpetrators/victims, the motives, and circumstances of the crime. The goal of the current study was to develop a better understanding of prison homicide by examining 54 perpetrators who committed 37 inmate homicides over 40 years in a mid-Western state prison system. Results showed that prison homicides typically involved a younger male inmate perpetrator, acting independently, murdering an older inmate, in his cell, by stabbing or beating the victim during an altercation. Perpetrators, in comparison with victims and prisoners in general, had a record indicating more prior community homicides, elevated institutional risk scores, and higher rates of serious and assaultive prison misconduct, all indicative of prior community and prison maladjustment. Consistent with career criminal research, prison homicide perpetrators constitute a small but distinct subset of habitually deviant criminals that perpetrate high rates of criminal and violent behavior regardless of context.

Diffusional motion as a gauge of fluidity and interfacial adhesion. Supported alkylphosphonate monolayers.

We report on the use of diffusion measurements to gauge the fluidity and surface binding properties of a molecular monolayer. The monolayer film consists of octadecyl-1-phosphonic acid (ODPA) and controlled amounts of a lyso-phosphatidic acid tagged with the fluorescent probe BODIPY (BLPA). The monolayer films were formed using a Langmuir-Blodgett (LB) trough and deposited onto a glass slide. Monolayer morphology was characterized during film formation using Brewster angle microscopy (BAM). Fluorescence Recovery After Photobleaching (FRAP) microscopy was used to measure translational diffusion of BLPA and Fluorescence Anisotropy Decay Imaging (FADI) was used to measure rotational diffusion of the BLPA chromophore. These results provide information on the motional freedom of the probe and, importantly, on the strength of interaction between the probe and the support. Compositional variations in the monolayer give rise to changes in constituent dynamics that reflect intermolecular interactions.

Testing the predictive validity of the Personality Assessment Inventory (PAI) in relation to inmate misconduct and violence.

The Personality Assessment Inventory (PAI) has been widely employed in correctional settings as a screening tool to assess inmates' risk for committing various types of institutional misconduct. Evaluations have generally found the PAI scales Antisocial Features (ANT), Aggression (AGG), and the Violence Potential Index (VPI) to be modestly related to institutional misbehavior, thus supporting its construct validity. The current study provides the most comprehensive examination of the predictive and incremental validity of the PAI and its subscales among a large sample of imprisoned offenders to date. In particular, the size of the sample (n = 15,546) and follow-up period (mean time at risk of 2.2 years) allowed for the disaggregation of institutional misconduct by levels of seriousness and separate examinations by conviction offense and criminal history variables. The 3 scales most strongly related to general rule infractions were ANT, AGG, and the VPI. After controlling for age at intake, violent conviction history, prior violent arrests, and time at risk, the PAI scales were shown to add incremental validity to the classification of 4 types of disciplinary infractions ranging from 2 to 4 percentage points. The study also explored the relationship of the PAI's response bias scales to institutional misconduct. (PsycINFO Database Record

An actuarial model for assessment of prison violence risk among maximum security inmates.

An experimental scale for the assessment of prison violence risk among maximum security inmates was developed from a logistic regression analysis involving inmates serving parole-eligible terms of varying length (n = 1,503), life-without-parole inmates (n = 960), and death-sentenced inmates who were mainstreamed into the general prison population (n = 132). Records of institutional violent misconduct of these 2,595 inmates were retrospectively examined for an 11-year period (1991 to 2002). Predictors affecting the likelihood of such misconduct included age, type and length of sentence, education, prior prison terms, prior probated sentences, and years served. The scale was modestly successful, as demonstrated by an overall omnibus area under the curve of .719. Double cross-validation demonstrated minimal shrinkage. The authors have termed this experimental scale the Risk Assessment Scale for Prison.

A single chain variant of factor VIII Fc fusion protein retains normal in vivo efficacy but exhibits altered in vitro activity.

Recombinant factor VIII Fc (rFVIIIFc) is a fusion protein consisting of a single B-domain-deleted (BDD) FVIII linked recombinantly to the Fc domain of human IgG1 to extend half-life. To determine if rFVIIIFc could be further improved by maintaining the heavy and light chains within a contiguous single chain (SC), we evaluated the activity and function of SC rFVIIIFc, an isoform that is not processed at residue R1648. SC rFVIIIFc showed equivalent activity in a chromogenic assay compared to rFVIIIFc, but approximately 40% activity by the one-stage clotting assay in the presence of von Willebrand Factor (VWF), with full activity in the absence of VWF. Moreover, SC rFVIIIFc demonstrated markedly delayed thrombin-mediated release from VWF, but an activity similar to that of rFVIIIFc upon activation in FXa generation assays. Therefore, the apparent reduction in specific activity in the aPTT assay appears to be primarily due to delayed release of FVIII from VWF. To assess whether stability and activity of SC rFVIIIFc were affected in vivo, a tail vein transection model in Hemophilia A mice was utilized. The results demonstrated similar pharmacokinetic profiles and comparable efficacy for SC rFVIIIFc and rFVIIIFc. Thus, while the single chain configuration did not promote enhanced half-life, it reduced the rate of release of FVIII from VWF required for activation. This impaired release may underlie the observed reduction in the one-stage clotting assay, but does not appear to affect the physiological activity of SC rFVIIIFc.

Pulmonary administration of interferon Beta-1a-fc fusion protein in non-human primates using an immunoglobulin transport pathway.

Currently, products containing interferon beta (IFNß) are injected either intramuscularly or subcutaneously. To avoid the necessity of injection, we developed a novel monomeric Fc fusion protein of IFNß (IFNßFc) that is absorbed via an immunoglobulin transport system present in the upper and central airways upon administration of the drug as an inhaled aerosol. The systemic absorption of IFNßFc through the lung in non-human primates, at deposited doses of 1, 3, and 10 µg/kg, was compared to the absorption of a single 3 µg/kg dose of IFNß-1a (Avonex®) subcutaneously administered. IFNßFc was well absorbed through the lung, displaying dose proportional increases in serum concentrations, and was biologically active, as shown by increases in plasma neopterin levels. The circulating half-life of IFNßFc was ∼3 times longer (∼30 h) than that of IFNß-1a, (8-9 h). At approximately equimolar doses of IFNßFc (10 µg/kg) and IFNß-1a (3 µg/kg), the stimulation of neopterin over background levels was approximately equivalent, demonstrating that the longer half-life of IFNßFc compensated for the lower relative specific antiviral activity of IFNßFc measured in vitro. In conclusion, IFNßFc was efficiently absorbed after pulmonary delivery in non-human primates, retained its biological activity, and may offer a convenient alternative to injectable IFNß.

CD74 deficiency ameliorates Pseudomonas aeruginosa-induced ocular infection.

Eye trauma and contact lens wear are the main factors that predispose to the development of infectious keratitis. The existing therapies fail to control the inflammation-driven tissue damage that occurs during Pseudomonas aeruginosa infection. Antibiotic treatment reduces bacterial burdens, but better interventions are needed to alleviate tissue damage resulting from local inflammation. We have previously documented that inhibition of macrophage migration inhibitory factor (MIF) reduces the bacterial levels and the inflammatory damage during keratitis. Here, we report that mice deficient for CD74, the putative MIF receptor, developed milder Pseudomonas aeruginosa-induced disease, characterized by decreased proinflammatory mediators and reduced bacterial presence in the cornea. However, topical inhibition of MIF using antibodies applied to the cornea further promoted recovery from disease, suggesting that in addition to MIF-dependent signaling events, MIF-triggered CD74-independent signaling pathways regulate sensitization to P. aeruginosa-induced infection.

Assertions of "future dangerousness" at federal capital sentencing: rates and correlates of subsequent prison misconduct and violence.

The federal prison disciplinary records of federal capital inmates (n=145) who were sentenced to life without possibility of release (LWOP) by plea bargain, pre-sentencing withdrawal of the death penalty, or jury determination were retrospectively reviewed (M=6.17 years post-admission). Disaggregated prevalence rates were inversely related to infraction severity: serious infraction =0.324, assaultive infraction =0.207, serious assault =0.09, assault with moderate injury =0.007, assault with major injuries or death =0.00. Frequency rates of misconduct were equivalent to other high-security federal inmates (n=18,561), regardless of infraction severity. Government assertions of "future dangerousness" as a nonstatutory aggravating factor were not predictive of prison misconduct. These findings inform federal capital risk assessments and have public policy implications for procedural reliability in death penalty prosecutions.

Is death row obsolete? A decade of mainstreaming death-sentenced inmates in Missouri.

Death-sentenced inmates in Missouri have been integrated or "mainstreamed" into the general population of the Potosi Correctional Center since 1991. By comparing the rate of violent misconduct among these mainstreamed death-sentenced inmates with that of the life-without-parole and parole eligible inmates under fully integrated conditions of confinement, this study provides the first empirical (statistical) evaluation of this innovative alternative to segregated death row confinement. The mainstreamed death-sentenced inmates committed no inmate or staff homicides, or attempted homicides. Comparison of their rates of institutional violence revealed frequencies that were similar to those of life-without-parole inmates, and well below those of fellow inmates who were sentenced to parole eligible terms. These findings cast serious doubt on the security-driven assumptions that have typified the segregation of death-sentenced inmates and have dictated highly restrictive confinement policies for this group. A conclusion that death-sentenced inmates can be safely integrated into a general prison population has significant implications for allocation of scarce fiscal resources and correctional staff, as well as for inmate mental health, particularly given the extended tenure that death-sentenced inmates typically serve between sentencing and relief/execution.