Cytomegalovirus variation among newborns treated with valganciclovir.

Congenital cytomegalovirus (cCMV) infection is the leading non-genetic cause of long-term neurological and sensory sequelae, the most common being sensorineural hearing loss (SNHL). Standard therapy for infants with symptomatic cCMV is valganciclovir for six months. However, little is known about the effects of antiviral therapy on CMV diversity while patients are on treatment. In this study, CMV variation was analyzed from urine specimens isolated from two patients with cCMV shortly after birth and at seven months. One was treated with valganciclovir for six weeks and the other for six months. In order to track these variants a novel bioinformatic approach was employed to analyze changes in low frequency variants over time. In the infant receiving antivirals for only six weeks, there was a fourfold increase in variation in UL97 from the seven month specimen. Furthermore, an eightfold increase in variation was seen in UL83 (pp65) with seven potential escape mutations occurring, and a twofold increase in UL73 (gN). In contrast variation did not increase or was reduced in these coding regions in the infant receiving valganciclovir for six months. However, there were increases in other CMV regions in samples isolated from both patients indicating further longitudinal studies are warranted to better understand the interplay between CMV diversity, antiviral therapy and patient outcome.

Correction to: Association of CMV genomic mutations with symptomatic infection and hearing loss in congenital CMV infection.

After publication of the original article [1], we were notified that Fig. 3 has "Fig. 1" posted on the top of it and Figs. 4 and 5 have "Genomic Position" in a different spot.

Association of CMV genomic mutations with symptomatic infection and hearing loss in congenital CMV infection.

BACKGROUND: Congenital cytomegalovirus (cCMV) infection is the most common congenital infection and a leading cause of long-term neurological and sensory sequelae, the most common being sensorineural hearing loss (SNHL). Despite extensive research, clinical or laboratory markers to identify CMV infected children with increased risk for disease have not been identified. This study utilizes viral whole-genome next generation-sequencing (NGS) of specimens from congenitally infected infants to explore viral diversity and specific viral variants that may be associated with symptomatic infection and SNHL. METHODS: CMV DNA from urine specimens of 30 infants (17 asymptomatic, 13 symptomatic) was target enriched and next generation sequenced resulting in 93% coverage of the CMV genome allowing analysis of viral diversity. RESULTS: Variant frequency distribution was compared between children with symptomatic and asymptomatic cCMV and those with (n = 13) and without (n = 17) hearing loss. The CMV genes UL48A, UL88, US19 and US22 were found to have an increase in nucleotide diversity in symptomatic children; while UL57, UL20, UL104, US14, UL115, and UL35 had an increase in diversity in children with hearing loss. An analysis of single variant differences between symptomatic and asymptomatic children found UL55 to have the highest number, while the most variants associated with SNHL were in the RL11 gene family. In asymptomatic infants with SNHL, mutations were observed more frequently in UL33 and UL20. CONCLUSION: CMV genomes from infected newborns can be mapped to 93% of the genome at a depth allowing accurate and reproducible analysis of polymorphisms for variant and gene discovery that may be linked to symptomatic and hearing loss outcomes.

Outbreak of Campylobacteriosis Associated with Raccoon Contact at a Wildlife Rehabilitation Centre, Minnesota, 2013.

Campylobacteriosis is an enteric illness caused by bacteria of the genus Campylobacter. There are approximately 900 culture-confirmed cases of campylobacteriosis reported annually to the Minnesota Department of Health (MDH). Case patients are interviewed about risk factors, including foods eaten, recreational and drinking water exposures and animal contact. In September 2013, MDH identified two Campylobacter jejuni cases who reported working at the same wildlife rehabilitation centre before illness onset. This report describes the investigation, which used a case-control study design, and identified 16 additional ill persons, for a total of 18 ill persons. Both cases and controls reported working with a variety of animals, including squirrels, chipmunks, mice, raccoons, opossums, rabbits, songbirds, waterfowl and reptiles. In univariate analyses, contact with a number of different animal species was significantly associated with illness, including raccoons (odds ratio [OR], 11.1; P < 0.001), chipmunks (OR, 3.65; P = 0.01), opossums (OR, 4.38; P = 0.005), mice (OR, 4.18; P = 0.01) and rabbits (OR, 4.36; P = 0.003). In a multivariate model, contact with raccoons was the only exposure independently associated with illness (adjusted OR, 12.2; P = 0.01). Bacterial culture and subtyping of the outbreak strain of C. jejuni from raccoon faecal samples further implicated raccoons as the source of the outbreak. Not all of the cases reported handling raccoons, suggesting that environmental contamination contributed to transmission. MDH worked with the wildlife rehabilitation centre's management to strengthen biosecurity and infection control protocols.

A Multi Targeting Conditionally Replicating Adenovirus Displays Enhanced Oncolysis while Maintaining Expression of Immunotherapeutic Agents.

Studies have demonstrated that oncolytic adenoviruses based on a 24 base pair deletion in the viral E1A gene (D24) may be promising therapeutics for treating a number of cancer types. In order to increase the therapeutic potential of these oncolytic viruses, a novel conditionally replicating adenovirus targeting multiple receptors upregulated on tumors was generated by incorporating an Ad5/3 fiber with a carboxyl terminus RGD ligand. The virus displayed full cytopathic effect in all tumor lines assayed at low titers with improved cytotoxicity over Ad5-RGD D24, Ad5/3 D24 and an HSV oncolytic virus. The virus was then engineered to deliver immunotherapeutic agents such as GM-CSF while maintaining enhanced heterogenic oncolysis.

alpha-Actinin interacts with rapsyn in agrin-stimulated AChR clustering.

AChR is concentrated at the postjunctional membrane at the neuromuscular junction. However, the underlying mechanism is unclear. We show that α-actinin, a protein known to cross-link F-actin, interacts with rapsyn, a scaffold protein essential for neuromuscular junction formation. α-Actinin, rapsyn, and surface AChR form a ternary complex. Moreover, the rapsyn-α-actinin interaction is increased by agrin, a factor known to stimulate AChR clustering. Downregulation of α-actinin expression inhibits agrin-mediated AChR clustering. Furthermore, the rapsyn-α-actinin interaction can be disrupted by inhibiting Abl and by cholinergic stimulation. Together these results indicate a role for α-actinin in AChR clustering.

The role of the cytoskeleton in neuromuscular junction formation.

The cytoskeleton plays a vital role in neuromuscular junction (NMJ) formation. It is responsible for shaping synaptic membrane into folds opposed to presynaptic active zones and anchoring acetylcholine receptors (AChRs) to the crest of the junctional folds. Acetylcholine receptors (AChRs) associate with the actin cytoskeleton, the disruption of which affects spontaneous and agrin-induced AChR clusters (Prives et al., 1982; Connolly, 1984; Peng and Phelan, 1984; Bloch, 1986; Dai et al., 2000). How AChRs are tethered to the actin cytoskeleton remains unclear.

Implication of geranylgeranyltransferase I in synapse formation.

Agrin activates the transmembrane tyrosine kinase MuSK to mediate acetylcholine receptor (AChR) clustering at the neuromuscular junction (NMJ). However, the intracellular signaling mechanism downstream of MuSK is poorly characterized. This study provides evidence that geranylgeranyltransferase I (GGT) is an important signaling component in the Agrin/MuSK pathway. Agrin causes a rapid increase in tyrosine phosphorylation of the alpha(G/F) subunit of GGT and in GGT activity. Inhibition of GGT activity or expression prevents muscle cells from forming AChR clusters in response to Agrin and attenuates the formation of neuromuscular synapses in spinal neuron-muscle cocultures. Importantly, transgenic mice expressing an alpha(G/F) mutant demonstrate NMJ defects with wider endplate bands and smaller AChR plaques. These results support the notion that prenylation is necessary for AChR clustering and the NMJ formation and/or maintenance, revealing an active role of GGT in Agrin/MuSK signaling.

Signaling complexes for postsynaptic differentiation.

The receptor tyrosine kinase MuSK is activated by agrin, an extracellular matrix protein believed to be utilized by motoneurons to regulate the formation or maintenance of the neuromuscular junction (NMJ). Recent studies have shed light on intracellular signaling mechanisms downstream of MuSK. Agrin enhances the activity of Rho GTPases and PAK, which is required for AChR clustering. Activation of these enzymes requires not only the kinase activity of MuSK, but also its interaction with proteins such as Dishevelled. These results suggest that MuSK may function as a scaffold tyrosine kinase that forms a multi-molecule complex for AChR clustering.

Urinalysis: current status and prospects for the future.

More than 300 million routine clinical analyses are performed annually in the United States. Methods for routine clinical urine examination, including detection of bacteriuria, are briefly reviewed. Prospects of some newer, better techniques to carry out such analyses are introduced. A preliminary report is presented on the use of supravital microscopic fluorescence technique (SMFT), employing acridine orange as a non-specific staining fluorochrome. Results of examining 218 unspun urine specimens by SMFT are compared to traditional bacteriologic culture at a large pediatric hospital reference laboratory.

Congenital cytomegalovirus disease in the Louisville area: a significant public health problem.

Using a passive surveillance system, 15 cases of symptomatic congenital cytomegalovirus (CMV) infection were identified in the Louisville area over approximately 3 years. When compared to mothers delivering healthy infants in Jefferson County, mothers of infants with congenital CMV were younger, more often nonwhite, and from lower socioeconomic status. Affected infants had a lower mean gestational age and birth weight, and most demonstrated classic features of congenital CMV infection such as jaundice, petechiae, hepatosplenomegaly, intrauterine growth retardation, and microcephaly. The overall infant mortality rate was 33%. Maternal and infant characteristics were similar to those of 167 cases in a national database and 106 cases reported from Birmingham, Alabama. The estimated minimal incidence of symptomatic congenital CMV disease in this area was 1.16/10,000 live births for white and 6.06/10,000 for nonwhites, establishing this as a significant public health problem in this part of Kentucky.