Herpes simplex virus type 1 glycoprotein gM and the membrane associated protein UL11 are required for virus-induced cell fusion and efficient virus entry

HSV-1 facilitates virus entry into cells and cell-to-cell spread by mediating fusion of the viral envelope with cellular membranes and fusion of adjacent cellular membranes. Although virus strains isolated from herpetic lesions cause limited cell fusion in cell culture, clinical herpetic lesions typically contain large syncytia, underscoring the importance of cell-to-cell fusion in virus spread in infected tissues. Certain mutations in gB, gK, UL20 and other viral genes, drastically enhance virus-induced cell fusion in vitro and in vivo. Recent work has suggested that gB is the sole fusogenic glycoprotein, which is regulated by interactions with viral glycoproteins gD, gH/gL, gK, membrane protein UL20 and cellular receptors. Recombinant viruses were constructed to abolish either gM or UL11 expression in the presence of strong syncytial mutations in either gB or gK. Virus-induced cell fusion caused by deletion of the carboxyl terminal 28 amino acids of gB, or the dominant syncytial mutation in gK (Ala-to-Val at amino acid 40), was drastically reduced in the absence of gM. Similarly, syncytial mutations in either gB or gK did not cause cell fusion in the absence of UL11. Neither the gM nor UL11 gene deletions substantially affected gB, gC, gD, gE and gH glycoprotein synthesis and expression on infected cell surfaces. Two-way immunoprecipitation experiments revealed that the membrane protein UL20, which is found as a protein complex with gK, interacted with gM, while gM did not interact with other viral glycoproteins. Viruses produced in the absence of gM or UL11 entered into cells slower than their parental wild-type virus. Moreover, the mouse eye model study, revealed that although the UL11-null virus replicated less efficiently in mouse corneas, it infected ganglionic neurons with greater efficiency than either the gM or gE-null viruses. Collectively, these results indicate that gM and UL11 are required for efficient membrane fusion events during virus entry and virus spread in vivo as well as in vitro, and also that gK plays the most important role in corneal and ganglionic infection in the mouse eye model followed by gM, gE, and UL11 in a descending order of importance relative to the wild-type virus

Measuring IS Service Quality in the Context of the Service Quality-User Satisfaction Relationship

There is little research regarding the relationship between IS service quality and user satisfaction, the most frequently used surrogate for information systems success. The current study is designed to investigate three ways of measuring service quality (i.e., confirmation/disconfirmation, perception-only, and overall assessment) and shed light on the relationship between service quality and user satisfaction. The results imply that when managers try to measure service quality to improve their service, they have to be cautious in ruling out or selecting one way or another of measuring service quality. The current research also clearly shows that mangers have to take care of the service quality to enhance user satisfaction. The models and results are discussed

Dual priming oligonucleotide system for the multiplex detection of respiratory viruses and SNP genotyping of CYP2C19 gene

Successful PCR starts with proper priming between an oligonucleotide primer and the template DNA. However, the inevitable risk of mismatched priming cannot be avoided in the currently used primer system, even though considerable time and effort are devoted to primer design and optimization of reaction conditions. Here, we report a novel dual priming oligonucleotide (DPO) which contains two separate priming regions joined by a polydeoxyinosine linker. The linker assumes a bubble-like structure which itself is not involved in priming, but rather delineates the boundary between the two parts of the primer. This structure results in two primer segments with distinct annealing properties: a longer 5′-segment that initiates stable priming, and a short 3′-segment that determines target-specific extension. This DPO-based system is a fundamental tool for blocking extension of non-specifically primed templates, and thereby generates consistently high PCR specificity even under less than optimal PCR conditions. The strength and utility of the DPO system are demonstrated here using multiplex PCR and SNP genotyping PCR

Optimization of magnetic flux density for fast MREIT conductivity imaging using multi-echo interleaved partial fourier acquisitions

BACKGROUND: Magnetic resonance electrical impedance tomography (MREIT) has been introduced as a non-invasive method for visualizing the internal conductivity and/or current density of an electrically conductive object by externally injected currents. The injected current through a pair of surface electrodes induces a magnetic flux density distribution inside the imaging object, which results in additional magnetic flux density. To measure the magnetic flux density signal in MREIT, the phase difference approach in an interleaved encoding scheme cancels out the systematic artifacts accumulated in phase signals and also reduces the random noise effect by doubling the measured magnetic flux density signal. For practical applications of in vivo MREIT, it is essential to reduce the scan duration maintaining spatial-resolution and sufficient contrast. In this paper, we optimize the magnetic flux density by using a fast gradient multi-echo MR pulse sequence. To recover the one component of magnetic flux density B(z), we use a coupled partial Fourier acquisitions in the interleaved sense. METHODS: To prove the proposed algorithm, we performed numerical simulations using a two-dimensional finite-element model. For a real experiment, we designed a phantom filled with a calibrated saline solution and located a rubber balloon inside the phantom. The rubber balloon was inflated by injecting the same saline solution during the MREIT imaging. We used the multi-echo fast low angle shot (FLASH) MR pulse sequence for MRI scan, which allows the reduction of measuring time without a substantial loss in image quality. RESULTS: Under the assumption of a priori phase artifact map from a reference scan, we rigorously investigated the convergence ratio of the proposed method, which was closely related with the number of measured phase encode set and the frequency range of the background field inhomogeneity. In the phantom experiment with a partial Fourier acquisition, the total scan time was less than 6 seconds to measure the magnetic flux density B(z) data with 128×128 spacial matrix size, where it required 10.24 seconds to fill the complete k-space region. CONCLUSION: Numerical simulation and experimental results demonstrated that the proposed method reduces the scanning time and provides the recovered B(z) data comparable to what we obtained by measuring complete k-space data

Molecular Detection of Dirofilaria immitis Specific Gene from Infected Dog Blood Sample Using Polymerase Chain Reaction

Background: Dirofilaria immitis, a filarial nematode, is the most important parasite-affecting dogs, causing cardiopulmonary dirofilariasis. Current diagnostic tools for detecting D. immitis include morphological assays, antigen detection, and X-ray. Herein, we developed a method for the molecular detection of D. immitis in blood using polymerase chain reaction (PCR). Methods: The study was conducted at Eulji University, Republic of Korea in 2016. To detect D. immitis-specific gene regions, we aligned the cytochrome c oxidase subunit I (COI) genes of seven filarial nematodes and designed primers targeting the unique region. We used dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-targeted primers as the internal control. We conducted PCR-amplified genomic DNA from canine blood samples. The products were confirmed by sequencing. Results: Gene alignment revealed a D. immitis COI-specific gene region, and the activity of designed primers was confirmed by PCR and sequencing. Plasmid DNA made from the PCR products was a positive control. The limit of detection for our method was 50 copies. The D. immitis COI and dog GAPDH genes could be discriminated from blood samples simultaneously. Conclusion: This study provides a method for highly specific and sensitive molecular diagnosis of D. immitis used as a diagnostic and therapeutic tool from the early stage of infection