An ultralong CDRH2 in HCV neutralizing antibody demonstrates structural plasticity of antibodies against E2 glycoprotein

A vaccine protective against diverse HCV variants is needed to control the HCV epidemic. Structures of E2 complexes with front layer-specific broadly neutralizing antibodies (bNAbs) isolated from HCV-infected individuals, revealed a disulfide bond-containing CDRH3 that adopts straight (individuals who clear infection) or bent (individuals with chronic infection) conformation. To investigate whether a straight versus bent disulfide bond-containing CDRH3 is specific to particular HCV-infected individuals, we solved a crystal structure of the HCV E2 ectodomain in complex with AR3X, a bNAb with an unusually long CDRH2 that was isolated from the chronically-infected individual from whom the bent CDRH3 bNAbs were derived. The structure revealed that AR3X utilizes both its ultralong CDRH2 and a disulfide motif-containing straight CDRH3 to recognize the E2 front layer. These results demonstrate that both the straight and bent CDRH3 classes of HCV bNAb can be elicited in a single individual, revealing a structural plasticity of VH1-69-derived bNAbs

Influences of Stacking Architectures of TiO 2

We investigated the influences of stacking architectures of the TiO2 nanoparticle layers on characteristics and performances of DSSCs. TiO2 nanoparticles of different sizes and compositions were characterized for their morphological and optical/scattering properties in thin films. They were used to construct different stacking architectures of the TiO2 nanoparticle layers for use as working electrodes of DSSCs. Characteristics and performances of DSSCs were examined to establish correlation of the stacking architectures of TiO2 nanoparticle layers with characteristics of DSSCs. The results suggest that the three-layer DSSC architecture, with sandwiching a 20 nm TiO2 nanoparticle layer between a 37 nm TiO2 nanoparticle layer and a hundred nm sized TiO2 back scattering/reflection layer, is effective in enhancing DSSC efficiencies. The high-total-transmittance 37 nm TiO2 nanoparticle layer with a larger haze can serve as an effective front scattering layer to scatter a portion of the incident light into larger oblique angles and therefore increase optical paths and absorption

Experience, age and exporting performance in UK SMEs

We consider the determinants of SME exporting performance using a survey of internationally engaged UK SMEs. We first develop a model incorporating organisational and prior managerial learning effects. Our empirical analysis then allows us to identify separately the positive effects on exporting from the international experience of the firm and the negative effects of firm age. Positive exporting effects also result from grafted knowledge – acquired by the recruitment of management with prior international experience. Innovation also has positive exporting effects with more radical new-to-the-industry innovation most strongly linked to inter-regional exports; new-to-the-firm innovation is more strongly linked to intra-regional trade. Early internationalisation is also linked positively to the number of countries to which firms export and the intensity of their export activity. We find no evidence, however, relating early internationalisation to extra-regional exporting, suggesting that early-exporting SMEs tend be ‘born regional’ rather than ‘born global’

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

The prevalence of rectal carriage of Klebsiella pneumoniae amongst diabetic patients and their clinical relevance in Taiwan: A five-year prospective study

Background/purpose: Pyogenic liver abscess (PLA) and bacteremia caused by Klebsiella pneumoniae is a common complication among patients with diabetes mellitus (DM). The aim of this study is to investigate the prevalence of rectal carriage and serotype distribution of K. pneumoniae amongst DM patients and their clinical relevance. Methods: We prospectively collected rectal swabs for K. pneumoniae culture in asymptomatic DM patients from March 2008 to June 2009. Seven capsular serotypes that were commonly associated with PLA were determined by capsular polysaccharide synthesis (cps) genotyping. Microbiologically confirmed bacterial infections were evaluated 1 and 5 years after initial enrolment of the patients. Results: A total of 100 male and 62 female patients (mean age, 56.6 years) were enrolled. Of these, 77 (47.5%) had rectal K. pneumoniae colonization. Colonizers were older than non-colonizers (p = 0.03). Sex, fasting blood glucose, and initial HbA1C were not statistically different (p = 0.26, 0.18, and 0.31, respectively). Among the 65 available isolates, 22 (33.8%) belonged to the seven main serotypes. During the 5-year's follow-up, 21 patients developed microbiologically documented bacterial infections but none of them developed PLA and bacteremia. Risk factors for bacterial infection within 5 years included initial glycosylated hemoglobin (HbA1C) > 10% or first-year average HbA1C > 10%. Conclusion: Although nearly half of asymptomatic DM patients had rectal carriage of K. pneumoniae and one-third of them colonized by isolates belonging to the seven serotypes related to PLA, none of them subsequently developed PLA and colonized patients did not have higher risk of microbiologically confirmed bacterial infections. Keywords: Klebsiella pneumoniae, Pyogenic liver abscess, Diabetes mellitus, Bacterial infections, Rectal carriag

Low-level laser irradiation stimulates tenocyte migration with up-regulation of dynamin II expression.

Low-level laser therapy (LLLT) is commonly used to treat sports-related tendinopathy or tendon injury. Tendon healing requires tenocyte migration to the repair site, followed by proliferation and synthesis of the extracellular matrix. This study was designed to determine the effect of laser on tenocyte migration. Furthermore, the correlation between this effect and expression of dynamin 2, a positive regulator of cell motility, was also investigated. Tenocytes intrinsic to rat Achilles tendon were treated with low-level laser (660 nm with energy density at 1.0, 1.5, and 2.0 J/cm(2)). Tenocyte migration was evaluated by an in vitro wound healing model and by transwell filter migration assay. The messenger RNA (mRNA) and protein expressions of dynamin 2 were determined by reverse transcription/real-time polymerase chain reaction (real-time PCR) and Western blot analysis respectively. Immunofluorescence staining was used to evaluate the dynamin 2 expression in tenocytes. Tenocytes with or without laser irradiation was treated with dynasore, a dynamin competitor and then underwent transwell filter migration assay. In vitro wound model revealed that more tenocytes with laser irradiation migrated across the wound border to the cell-free zone. Transwell filter migration assay confirmed that tenocyte migration was enhanced dose-dependently by laser. Real-time PCR and Western-blot analysis demonstrated that mRNA and protein expressions of dynamin 2 were up-regulated by laser irradiation dose-dependently. Confocal microscopy showed that laser enhanced the expression of dynamin 2 in cytoplasm of tenocytes. The stimulation effect of laser on tenocytes migration was suppressed by dynasore. In conclusion, low-level laser irradiation stimulates tenocyte migration in a process that is mediated by up-regulation of dynamin 2, which can be suppressed by dynasore

Transwell filter migration assay revealed that laser stimulated tenocytes migration <i>in vitro</i> (*indicates <i>p</i><0.05 between laser-treated and control tenocytes).

<p>Transwell filter migration assay revealed that laser stimulated tenocytes migration <i>in vitro</i> (*indicates <i>p</i><0.05 between laser-treated and control tenocytes).</p