Dual infection of novel influenza viruses A/H1N1 and A/H3N2 in a cluster of Cambodian patients.

During the early months of 2009, a novel influenza A/H1N1 virus (pH1N1) emerged in Mexico and quickly spread across the globe. In October 2009, a 23-year-old male residing in central Cambodia was diagnosed with pH1N1. Subsequently, a cluster of four influenza-like illness cases developed involving three children who resided in his home and the children's school teacher. Base composition analysis of internal genes using reverse transcriptase polymerase chain reaction and electrospray ionization mass spectrometry revealed that specimens from two of the secondary victims were coinfected with influenza A/H3N2 and pH1N1. Phylogenetic analysis of the hemagglutinin genes from these isolated viruses showed that they were closely related to existing pH1N1 and A/H3N2 viruses circulating in the region. Genetic recombination was not evident within plaque-purified viral isolates on full genome sequencing. This incident confirms dual influenza virus infections and highlights the risk of zoonotic and seasonal influenza viruses to coinfect and possibly, reassort where they cocirculate.

Chlamydophila pneumoniae infection among Basic Underwater Demolition/SEAL (BUD/S) candidates, Coronado, California, July 2008.

Community-acquired pneumonia can compromise readiness of recruits and service members operating in confined spaces. Often respiratory pathogens are implicated in outbreaks. In July 2008, 5 Basic Underwater Demolition/SEAL students entering an intense period of training at Naval Amphibious Base Coronado reported with clinical symptoms and chest radiographs consistent with pneumonia. Throat and nasal swabs were tested for respiratory pathogens. Molecular evidence indicated that they were infected with the atypical bacterium Chlamydophila pneumoniae. Thirty contemporaneous Basic Underwater Demolition/SEAL students were tested to determine the extent of C pneumoniae infection burden. Five additional cases were captured within this group. The 10 individuals diagnosed with C pneumoniae were treated with a course of azithromycin, Avelox (moxifloxacin hydrochloride), and doxycycline. The cases ended following the isolation of cases and prophylaxis with oral antibiotics. This work highlights the importance of rapid respiratory disease diagnoses to guide the clinical response following the emergence of respiratory infections among military trainees.

A growing global network's role in outbreak response: AFHSC-GEIS 2008-2009.

A cornerstone of effective disease surveillance programs comprises the early identification of infectious threats and the subsequent rapid response to prevent further spread. Effectively identifying, tracking and responding to these threats is often difficult and requires international cooperation due to the rapidity with which diseases cross national borders and spread throughout the global community as a result of travel and migration by humans and animals. From Oct.1, 2008 to Sept. 30, 2009, the United States Department of Defense's (DoD) Armed Forces Health Surveillance Center Global Emerging Infections Surveillance and Response System (AFHSC-GEIS) identified 76 outbreaks in 53 countries. Emerging infectious disease outbreaks were identified by the global network and included a wide spectrum of support activities in collaboration with host country partners, several of which were in direct support of the World Health Organization's (WHO) International Health Regulations (IHR) (2005). The network also supported military forces around the world affected by the novel influenza A/H1N1 pandemic of 2009. With IHR (2005) as the guiding framework for action, the AFHSC-GEIS network of international partners and overseas research laboratories continues to develop into a far-reaching system for identifying, analyzing and responding to emerging disease threats.

Outbreak of acute respiratory disease caused by Mycoplasma pneumoniae on board a deployed U.S. navy ship.

We identified 179 cases of acute respiratory illness including 50 cases of radiographically confirmed pneumonia over the course of 4 months on a deployed U.S. Navy vessel. Laboratory tests showed Mycoplasma pneumoniae to be the etiological agent. This report represents the first published description of a shipboard outbreak of this pathogen.

Diagnostic discrimination of live attenuated influenza vaccine strains and community-acquired pathogenic strains in clinical samples.

Live vaccines can generate false-positive results on common influenza assays including reverse transcriptase-PCR (RT-PCR), culture and antigen tests. This threatens the integrity of epidemiological data and may misdirect treatment and control efforts. We report the development of RT-PCR tests that distinguish live FluMist vaccine (FMV) strains from circulating influenza strains in clinical samples. Primers were validated using influenza-positive samples from unvaccinated patients, packaged FMV, and one PCR-positive asymptomatic vaccine. Furthermore, the assay was used to experimentally test our lab's collection of influenza-positive samples from the 2004-05 and 2005-06 influenza seasons and several 2005 preseason isolates to determine the rate of vaccine-derived false-positive results under differing epidemiological conditions. Analytical and clinical validations show that the assay is both sensitive and specific. Experimental results demonstrate that 51 out of 51 influenza-positive samples collected during influenza season from ill, previously-vaccinated military personnel represent real infections with circulating strains. Finally, the assay shows that four preseason influenza-positive samples were false positives resulting from vaccine shedding. The vaccine-discriminatory RT-PCR methods described here provide the first test designed to distinguish FMV strains from circulating strains. The results show that the test is effective, and demonstrate the importance of such tests in the age of live vaccines.

Retargeting mobile group II introns to repair mutant genes.

Retroposable elements such as retroviral and lentiviral vectors have been employed for many gene therapy applications. Unfortunately, such gene transfer vectors integrate genes into many different DNA sequences and unintended integration of the vector near a growth-promoting gene can engender pathological consequences. For example, retroviral vector-mediated gene transfer induced leukemia in 2 of 11 children treated for severe combined immunodeficiency, raising significant safety issues for gene transfer strategies that cannot be targeted to specific sequences. Here, we examine the use of a mobile retroposable genetic element that can be targeted to introduce therapeutic sequences site specifically into mutant genes. The data demonstrate that the mobile group II intron from Lactococcus lactis can be targeted to insert into and repair mutant lacZ (approved gene symbol GLB1) and beta-globin (approved gene symbol HBB) genes with high efficiency and fidelity in model systems in bacteria. These results suggest that these mobile genetic elements represent a novel class of agents for performing targeted genetic repair.

A tyrosyl-tRNA synthetase adapted to function in group I intron splicing by acquiring a new RNA binding surface.

We determined a 1.95 A X-ray crystal structure of a C-terminally truncated Neurospora crassa mitochondrial tyrosyl-tRNA synthetase (CYT-18 protein) that functions in splicing group I introns. CYT-18's nucleotide binding fold and intermediate alpha-helical domains superimpose on those of bacterial TyrRSs, except for an N-terminal extension and two small insertions not found in nonsplicing bacterial enzymes. These additions surround the cyt-18-1 mutation site and are sites of suppressor mutations that restore splicing, but not synthetase activity. Highly constrained models based on directed hydroxyl radical cleavage assays show that the group I intron binds at a site formed in part by the three additions on the nucleotide binding fold surface opposite that which binds tRNATyr. Our results show how essential proteins can progressively evolve new functions.

Misconceptions, discrimination, and disabling language: synthesis and review.

Misconceptions about various disabling conditions abound in the general population and are related to the presence of socially disabling stereotyping as a cultural phenomenon. Significant levels of misconception have been demonstrated among the population regarding head injuries, epilepsy, and electroconvulsive shock treatment. Public ignorance in these domains is thought to breed a climate ripe for discrimination, and numerous studies have shown that such discrimination does occur in the workplace and, perhaps more important, in the world of everyday discourse. Discrimination patterns are not uniform, however, and all disabling conditions are not alike in their susceptibility to discrimination. This article presents a review of the relationships among misconceptions, employment discrimination, and language discrimination patterns, as these factors function as obstacles to community reentry for many persons with disabilities. Strategies for overcoming the effects of social stereotyping and discrimination are offered, with the goal of educating the professional and rehabilitation communities about unwitting discrimination perpetrated by individuals belonging to these communities.