Rational Engineering and Preclinical Evaluation of Neddylation and SUMOylation Site Modified Adeno-Associated Virus Vectors in Murine Models of Hemophilia B and Leber Congenital Amaurosis.

Synthetic engineering of viral vectors such as adeno-associated virus (AAV) is crucial to overcome host transduction barriers observed during clinical gene therapy. We reasoned that exploring the role of cellular ubiquitin-like modifiers (UBLs) such as Neddylation or SUMOylation during AAV transduction could be beneficial. Using a combination of biochemical and molecular engineering strategies, we have studied the impact of these UBLs during AAV2 infection and further developed Neddylation or SUMOylation site-modified AAV vectors and validated them in multiple disease models and .

Analysis of hepatic and retinal cell microRNAome during AAV infection reveals their diverse impact on viral transduction and cellular physiology.

Cellular microRNAs are known to modulate the life-cycle of different viruses. Surprisingly, very little data exists on AAV-induced changes to the cellular microRNAome in general and in hepatic and retinal cells, in particular. We reasoned that inducible microRNA in response to recombinant AAV infection may regulate immediate and long-lived cellular responses necessary for the cell's own survival as well as its ability to control several aspects of viral life-cycle.

Molecular Engineering of Adeno-Associated Virus Capsid Improves Its Therapeutic Gene Transfer in Murine Models of Hemophilia and Retinal Degeneration.

Recombinant adeno-associated virus (AAV)-based gene therapy has been promising, but several host-related transduction or immune challenges remain. For this mode of therapy to be widely applicable, it is crucial to develop high transduction and permeating vectors that infect the target at significantly low doses. Because glycosylation of capsid proteins is known to be rate limiting in the life cycle of many viruses, we reasoned that perturbation of glycosylation sites in AAV2 capsid will enhance gene delivery.

Post-translational modifications in capsid proteins of recombinant adeno-associated virus (AAV) 1-rh10 serotypes.

Post-translational modifications in viral capsids are known to fine-tune and regulate several aspects of the infective life cycle of several viruses in the host. Recombinant viruses that are generated in a specific producer cell line are likely to inherit unique post-translational modifications during intra-cellular maturation of its capsid proteins. Data on such post-translational modifications in the capsid of recombinant adeno-associated virus serotypes (AAV1-rh10) is limited.

Visualization of Retinal Morphology in rd12 Mice by Scanning Electron Microscopy.

We have evaluated the retinal morphological changes in normal C57BL6/J mice and a retinal degenerative disease (rd12 mice) model using scanning electron microscopy. A method of sample preparation for electron microscopy was developed and the cryosectioned retina was used to study the retinal thickness. Our data demonstrate variation in the neural retina texture between rd12 mice and C57BL6/J mice.

Optimized AAV rh.10 Vectors That Partially Evade Neutralizing Antibodies during Hepatic Gene Transfer.

Of the 12 common serotypes used for gene delivery applications, Adeno-associated virus (AAV)rh.10 serotype has shown sustained hepatic transduction and has the lowest seropositivity in humans. We have evaluated if further modifications to AAVrh.

Single-center study of interrater agreement in the identification of central line-associated bloodstream infection.

Background: Interrater reliability of central line-associated bloodstream infection (CLABSI) determination has not been well studied. The present study evaluated interrater reliability between infection preventionists (IPs) for CLABSI- and other bloodstream infection (BSI)-related factors and examined whether any nurse characteristics are associated with interrater reliability.

Methods: A total of 165 blood cultures were reviewed by 2 IPs assigned at random.

Impact of isolation on hospital consumer assessment of healthcare providers and systems scores: is isolation isolating?

The Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) survey was used to measure the effect of isolation on patient satisfaction. Isolated patients reported lower scores for questions regarding physician communication and staff responsiveness. Overall scores for these domains were lower in isolated than in nonisolated patients.

Absence of an association between glucose levels and surgical site infections in patients undergoing craniotomies for brain tumors.

Object: In select patient populations, hyperglycemia has been shown to increase the risk of surgical site infection (SSI), whereas stringent glucose control has improved outcomes. To date, no study has focused on whether SSIs in patients with brain tumors undergoing resection are associated with hyperglycemia.

Methods: The authors performed a retrospective chart review of patients who underwent a craniotomy after receiving a diagnosis of brain tumor.

Novel use of the intranet to document health care personnel participation in a mandatory influenza vaccination reporting program.

Background: The Advisory Committee on Immunization Practices and the Healthcare Infection Control Practices Advisory Committee recommend health care personnel (HCP) influenza vaccination to prevent transmission to patients. Recommendations include documenting declination and providing feedback of rates. We report the use of the intranet to achieve recommendations with mandatory participation of 20,170 HCP in an 1100-bed hospital.

Outbreak of methicillin-resistant Staphylococcus aureus colonization and infection in a neonatal intensive care unit epidemiologically linked to a healthcare worker with chronic otitis.

Objective: To describe the investigation and interventions necessary to contain an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) colonization and infection in a neonatal intensive care unit (NICU).

Design: Retrospective case finding that involved prospective performance of surveillance cultures for detection of MRSA and molecular typing of MRSA by repetitive-sequence polymerase chain reaction (rep-PCR).

Setting: Level III NICU in a tertiary care center.