Phase IIa, randomised, double-blind study of GSK3389404 in patients with chronic hepatitis B on stable nucleos(t)ide therapy.

BACKGROUND & AIMS: Bepirovirsen, an antisense oligonucleotide targeting pregenomic and mRNA transcripts of HBV, has been conjugated to N-acetyl galactosamine (GSK3389404) to enhance hepatocyte delivery. This dose-finding study was the first to assess GSK3389404 for chronic HBV infection. METHODS: This phase IIa, randomised, double-blind, placebo-controlled, 2-part study was conducted in 22 centres in Asia (NCT03020745). Pharmacokinetic findings from Part 1 informed Part 2 dosing. In Part 2, patients with chronic hepatitis B on nucleos(t)ide analogue therapy were randomised 11:2 to GSK3389404 (30, 60, 120 mg weekly or 120 mg bi-weekly) or placebo until Day 85. Coprimary endpoints included HBsAg response (≥1.5 log10 IU/ml reduction from baseline) rate, safety and pharmacokinetics. RESULTS: Parts 1 and 2 included 12 (9 GSK3389404, 3 placebo) and 66 patients (56 GSK3389404, 10 placebo), respectively. In Part 2, one patient each in the 60 mg weekly, 120 mg weekly and 120 mg bi-weekly arms achieved a HBsAg response. HBsAg reductions were dose-dependent (Day 85: mean 0.34 [60 mg weekly] to 0.75 log10 IU/ml [120 mg weekly]) and occurred in hepatitis B e antigen-positive and -negative patients. No patient achieved HBsAg seroclearance. 43/56 (77%) GSK3389404- and 9/10 (90%) placebo-treated patients reported adverse events. No deaths were reported. Alanine aminotransferase flares (>2x upper limit of normal) occurred in 2 GSK3389404-treated patients (120 mg weekly, 120 mg bi-weekly); both were associated with decreased HBsAg, but neither was considered a responder. GSK3389404 plasma concentrations peaked 2-4 hours post dose; mean plasma half-life was 3-5 hours. CONCLUSIONS: GSK3389404 showed an acceptable safety profile and target engagement, with dose-dependent reductions in HBsAg. However, no efficacious dosing regimen was identified. CLINICAL TRIAL NUMBER: NCT03020745. LAY SUMMARY: Hepatitis B virus (HBV) can result in chronic HBV infection, which may ultimately lead to chronic liver disease, primary liver cancer and death; HBV proteins may prevent the immune system from successfully controlling the virus. GSK3389404 is an investigational agent that targets HBV RNA, resulting in reduced viral protein production. This study assessed the safety of GSK3389404 and its ability to reduce the viral proteins in patients with chronic HBV infection. GSK3389404 showed dose-dependent reduction in hepatitis B surface antigen, with an acceptable safety profile. While no clear optimal dose was identified, the findings from this study may help in the development of improved treatment options for patients with chronic HBV infections.

Design of structural coloration for full-color high-definition computer-generated holograms.

This paper investigates the submicron scale color filter design in the high-definition computer-generated hologram (HD-CGH). It is addressed that single pixel structural coloration is essential for full-color wide-viewing angle HD-CGH because the conventional RGB color stripe filter degrades HD-CGH image quality due to low misalignment tolerance. Considering that a submicron scale slit or hole with metallic mirror sidewalls can operate as a single pixel color filter. We propose a design of single pixel RGB plasmonic color filter (PCF) and present the feasibility of applying the proposed single pixel RGB PCF to high-definition HD-CGHs. Based on the RGB PCF platform, a 1.1 µm × 1.1 µm RGB PCF is designed and the corresponding optical characteristics of the full-color HD-CGH are analyzed.

Smoothing of inter-layer edge artifacts in depth-map computer-generated holograms.

In the depth-map computer-generated hologram (CGH), inter-layer edge artifacts are observed in the discontinuous edges of section-wise depth-map objects. CGH synthesis, utilizing the hybrid smoothing method of silhouette masking and edge-apodization, alleviates unwanted inter-layer edge artifacts. The proposed method achieves improved de-artifact filtering that generates holographic images closer to the ground truth image of the depth-map object unattainable by the conventional CGH synthesis method.

Charge Modulation Layer and Wide-Color Tunability in a QD-LED with Multiemission Layers.

Widely tunable color emission from a single pixel is a promising but challenging technology for quantum-dot light-emitting diodes (QD-LEDs). Even a QD-LED pixel with stacked multi-QD layers having different colors is likely to emit a monotonic color because the exciton recombination mostly occurs in 1 or 1.5 QD layers with better charge balance. In this study, an all-solution-processed QD-LED with electrically tunable color emission over a wide color range by introducing a charge modulation layer (CML) is developed. Specifically, the CML acted as a high and narrow energy barrier for electrons between two QD layers, and the electron drift is sensitively controlled via the field-dependent tunneling effect. Therefore, the charge distribution and balance in the two QD layers re-electrically tunable, which enhanced the color tunability. The color tuning range and quantum efficiency are effectively controlled depending on the CML material and thickness. In addition, the color change caused by the solvent effect in a QD-LED with dual QD layers is thoroughly investigated. The proposed method may advance the understanding of QD emission behavior with the use of CML and provide a practical approach for the actual application of color-tunable pixel technology.

Wide-viewing full-color depthmap computer-generated holograms.

An efficient synthesis algorithm for wide-viewing full-color depthmap computer-generated holograms is proposed. We develop a precise computational algorithm integrating wave-optic geometry-mapping, color-matching, and noise-filtering to multiplex multiview elementary computer-generated holograms (CGHs) into a single high-definition CGH without three-dimensional perspective distortion or color dispersion. Computational parallelism is exploited to achieve significant computational efficiency improvement in the production throughput of full-color wide-viewing angle CGHs. The proposed algorithm is verified through the full-color binary hologram reconstruction experiments utilizing an off-axis R·G·B simultaneous illumination method, which suggests the feasibility of the full-color sub-wavelength binary spatial light modulator technology.

Characteristics of complex light modulation through an amplitude-phase double-layer spatial light modulator.

The complex modulation characteristics of a light field through an amplitude-phase double-layer spatial light modulator are analyzed based on the wave-optic numerical model, and the structural conditions for the optimal double-layer complex modulation structure are investigated. The relationships of interlayer distance, pixel size, and complex light modulation performance are analyzed. The main finding of this study is that the optimal interlayer distance for the double-layer structure can be found at the Talbot effect condition. For validating the practical usefulness of our findings, a high quality reconstruction of the complex computer-generated holograms and the robustness of the angular tolerance of the complex modulation at the Talbot interlayer distance are numerically demonstrated.

Transcriptional regulation of IFN-λ genes in hepatitis C virus-infected hepatocytes via IRF-3·IRF-7·NF-κB complex.

Hepatitis C virus (HCV) infection in hepatocytes stimulates innate antiviral responses including the production of type III interferons (IFN-λ), including IL-28A, IL-28B, and IL-29. However, the molecular mechanism(s) regulating the expression of IFN-λ genes in HCV-infected hepatocytes remains undefined. In this study, we examined regulatory elements involved in the induction of IFN-λ genes following HCV infection in hepatocytes and further determined the binding of specific transcription factor(s) to promoter regions of IFN-λ genes. Our studies reveal that the regulatory portion for IL-28A, IL-28B, and IL-29 genes is localized to a 1-kb region in their respective promoters. Notably, interferon regulatory factor (IRF)-3 and -7 are the key transcriptional factors for the induction of IL-28A and IL-28B genes, whereas NF-κB is an additional requirement for the induction of the IL-29 gene. Ligation of Toll-like receptors (TLR) 3, 7, 8, and 9, which also activate IRFs and NF-κB, resulted in more robust production of IFN-λ than that observed with HCV infection, verifying the importance of TLR pathways in IFN-λ production. Furthermore, the addition of IFN-λ to HCV-infected hepatocytes decreased viral replication and produced a concurrent reduction in microRNA-122 (miR-122). The decrease in viral replication was enhanced by the co-administration of IFN-λ and miR-122 inhibitor (miRIDIAN), suggesting that such combinatorial therapies may be beneficial for the treatment of chronic HCV infection.

Exploring the spatio-temporal clusters of closed restaurants after the COVID-19 outbreak in Seoul using relative risk surfaces.

This study explores the clusters of closed restaurants in Seoul in response to the COVID-19 pandemic using the relative risk surface (RRS). The RRS developed based on kernel density estimation provides alternative perspectives for finding the cluster by combining different control and case events. Specifically, the varying impacts on diverse types of restaurants are examined by comparing the densities of closed casual restaurants and cafes. The clusters of closed businesses following the COVID-19 outbreak are subsequently explored through a comparison of the densities of the closed businesses preceding the outbreak. Furthermore, this analysis estimates the clusters of declined commercial areas after the pandemic outbreak based on the comparison between the densities of opened and closed restaurants. Finally, the specific time and region of the clusters are explored using space-time RRS. The analysis results effectively demonstrate various aspects of the closed restaurant clusters. For example, in the central business areas, the densities of closed cafes have decreased after the pandemic outbreak, and the density of closed cafes is significantly higher than that of opened cafes. This study would contribute to the literature on spatial data analysis and urban policy support in response to future epidemics.

Inverse Propensity Score-Weighted Analysis of Entecavir and Tenofovir Disoproxil Fumarate in Patients with Chronic Hepatitis B: A Large-Scale Multicenter Study.

Tenofovir disoproxil fumarate (TDF) is reportedly superior or at least comparable to entecavir (ETV) in preventing hepatocellular carcinoma (HCC) among chronic hepatitis B (CHB) patients; however, it remains controversial. This study aimed to conduct comprehensive comparisons between the two antivirals. CHB patients initially treated with ETV or TDF between 2012 and 2015 at 20 referral centers in Korea were included. The primary outcome was the cumulative incidence of HCC. The secondary outcomes included death or liver transplantation, liver-related outcome, extrahepatic malignancy, development of cirrhosis, decompensation events, complete virologic response (CVR), seroconversion rate, and safety. Baseline characteristics were balanced using the inverse probability of treatment weighting (IPTW). Overall, 4210 patients were enrolled: 1019 received ETV and 3191 received TDF. During the median follow-ups of 5.6 and 5.5 years, 86 and 232 cases of HCC were confirmed in the ETV and TDF groups, respectively. There was no difference in HCC incidence between the groups both before (p = 0.36) and after IPTW was applied (p = 0.81). Although the incidence of extrahepatic malignancy was significantly higher in the ETV group than in the TDF group before weighting (p = 0.02), no difference was confirmed after IPTW (p = 0.29). The cumulative incidence rates of death or liver transplantation, liver-related outcome, new cirrhosis development, and decompensation events were also comparable in the crude population (p = 0.24-0.91) and in the IPTW-adjusted population (p = 0.39-0.80). Both groups exhibited similar rates of CVR (ETV vs. TDF: 95.1% vs. 95.8%, p = 0.38), and negative conversion of hepatitis B e antigen (41.6% vs. 37.2%, p = 0.09) or surface antigen (2.8% vs. 1.9%, p = 0.10). Compared to the ETV group, more patients in the TDF group changed initial antivirals due to side effects, including decreased kidney function (n = 17), hypophosphatemia (n = 20), and osteoporosis (n = 18). In this large-scale multicenter study, ETV and TDF demonstrated comparable effectiveness across a broad range of outcomes in patients with treatment-naïve CHB during similar follow-up periods.

Safety, tolerability and antiviral activity of the antisense oligonucleotide bepirovirsen in patients with chronic hepatitis B: a phase 2 randomized controlled trial.

Chronic infection with hepatitis B virus (HBV) leads to an increased risk of death from cirrhosis and hepatocellular carcinoma. Functional cure rates are low with current treatment options (nucleos(t)ide analogs (NAs) and pegylated interferons). Bepirovirsen is an antisense oligonucleotide targeting all HBV messenger RNAs; in cell culture and animal models, bepirovirsen leads to reductions in HBV-derived RNAs, HBV DNA and viral proteins. This phase 2 double-blinded, randomized, placebo-controlled trial is the first evaluation of the safety and activity of an antisense oligonucleotide targeting HBV RNA in both treatment-naïve and virally suppressed individuals with chronic HBV infection. The primary objective was to assess the safety and tolerability of bepirovirsen in individuals with chronic hepatitis B (CHB) (NCT02981602). The secondary objective was to assess antiviral activity, including the change from baseline to day 29 in serum hepatitis B surface antigen (HBsAg) concentration. Participants with CHB infection ≥6 months and serum HBsAg ≥50 IU ml-1 were enrolled from seven centers across Hong Kong and the Republic of Korea and randomized (3:1 within each dose cohort) to receive bepirovirsen or placebo via subcutaneous injection twice weekly during weeks 1 and 2 (days 1, 4, 8 and 11) and once weekly during weeks 3 and 4 (days 15 and 22). Participants were then followed for 26 weeks. Twenty-four participants were treatment-naïve and seven were receiving stable NA therapy. Treatment-emergent adverse events were mostly mild/moderate (most commonly injection site reactions). Eleven (61.1%) and three (50.0%) treatment-naïve participants experienced one or more treatment-emergent adverse event in the bepirovirsen and placebo groups, respectively. In participants receiving NA therapy, the corresponding numbers were three (60.0%) and one (50.0%). Transient, self-resolving alanine aminotransferase flares (≥2× upper limit of normal) were observed in eight treatment-naïve participants and three participants on stable NA regimens in the bepirovirsen treatment arms. HBsAg reductions were observed and were significant versus placebo for treatment-naïve participants receiving bepirovirsen 300 mg (P = 0.001), but not for the bepirovirsen 150 mg group (P = 0.245) or participants receiving stable NA therapy (P = 0.762). Two participants in each of the 300 mg dose groups achieved HBsAg levels below the lower limit of quantitation by day 29 (n = 3) or day 36 (n = 1). Bepirovirsen had a favorable safety profile. These preliminary observations warrant further investigation of the safety and activity of bepirovirsen in a larger CHB patient population.

Photothermally Modulatable and Structurally Disintegratable Sub-8-nm Au1Ag9 Embedded Nanoblocks for Combination Cancer Therapy Produced by Plug-in Assembly.

As well as the exploration of translatable delivery nanosystems for cancer therapeutic agents, the development of automatable continuous-flow manufacturing technology comprising digitally controlled reactions for the on-demand production of pharmaceuticals is an important challenge in anticancer nanomedicine. Most attempts to resolve these issues have involved the development of alternative reactions, formulations, or constructs containing stimulus components aimed at producing multiple approaches for highly efficacious combination cancer therapies. However, there has been no report of a platform based on plug-in execution that enables continuous-flow manufacture in a compact, reconfigurable manner, although an optimal platform technology may be a prerequisite for the timely translation of recently developed nanomedicines. To this end, we describe the development of a platform toward digitizable, continuous manufacture by a serial combination of plug-in reactionwares (heating plates, a spraying cup, and a photochamber) for single-pass flow fabrication. Specifically, we fabricated three different composite nanoblocks consisting of Au1Ag9 (<8 nm; stimulus component), docetaxel (an anticancer drug), and bovine serum albumin (a protective and targeting agent) using our system, with the result of producing nanoblocks with photothermally modulatable and structurally disintegratable properties. These were examined for effectiveness in near-infrared-induced chemothermal cancer therapy and renal excretion of Au1Ag9 particles and exhibited high anticancer efficacy and warrantable biosafety.

Biomechanical analysis of a newly developed interspinous process device conjunction with interbody cage based on a finite element model.

PURPOSE: This study aimed to investigate the biomechanical effects of a newly developed interspinous process device (IPD), called TAU. This device was compared with another IPD (SPIRE) and the pedicle screw fixation (PSF) technique at the surgical and adjacent levels of the lumbar spine. MATERIALS AND METHODS: A three-dimensional finite element model analysis of the L1-S1 segments was performed to assess the biomechanical effects of the proposed IPD combined with an interbody cage. Three surgical models-two IPD models (TAU and SPIRE) and one PSF model-were developed. The biomechanical effects, such as range of motion (ROM), intradiscal pressure (IDP), disc stress, and facet loads during extension were analyzed at surgical (L3-L4) and adjacent levels (L2-L3 and L4-L5). The study analyzed biomechanical parameters assuming that the implants were perfectly fused with the lumbar spine. RESULTS: The TAU model resulted in a 45%, 49%, 65%, and 51% decrease in the ROM at the surgical level in flexion, extension, lateral bending, and axial rotation, respectively, when compared to the intact model. Compared to the SPIRE model, TAU demonstrated advantages in stabilizing the surgical level, in all directions. In addition, the TAU model increased IDP at the L2-L3 and L4-L5 levels by 118.0% and 78.5% in flexion, 92.6% and 65.5% in extension, 84.4% and 82.3% in lateral bending, and 125.8% and 218.8% in axial rotation, respectively. Further, the TAU model exhibited less compensation at adjacent levels than the PSF model in terms of ROM, IDP, disc stress, and facet loads, which may lower the incidence of the adjacent segment disease (ASD). CONCLUSION: The TAU model demonstrated more stabilization at the surgical level than SPIRE but less stabilization than the PSF model. Further, the TAU model demonstrated less compensation at adjacent levels than the PSF model, which may lower the incidence of ASD in the long term. The TAU device can be used as an alternative system for treating degenerative lumbar disease while maintaining the physiological properties of the lumbar spine and minimizing the degeneration of adjacent segments.

All-solution-processed colour-tuneable tandem quantum-dot light-emitting diode driven by AC signal.

We demonstrate a novel structure for a quantum-dot light-emitting diode (QD-LED) with wide-range colour-tuneable pixels, fabricated via full solution processing. The proposed device has a symmetrical structure produced via stacking of an inverted-structure diode with a green QD emission layer (EML) and normal-structure diode with a red QD EML. It is an electron-only device; however, a charge generation layer in the middle of the device generates holes for the formation of excitons. Depending on the polarity of the applied voltage, either the bottom inverted unit or the top normal unit is operated, thereby emitting green or red light, respectively. The working mechanism of the device is investigated via analysis of the charge generation mechanism and carrier transport path. In addition, the colour tunability is verified using a simple alternating current (AC) driving scheme; the duty cycle modulation of the AC signal enables fine colour adjustment over a broad range, from pure green to pure red. Thus, our colour-tuneable QD-LED with vertically stacked independently operated sub-pixels can open a promising pathway towards cost-effective ultra-high-resolution displays.

Blocking of monocyte-associated B7-H1 (CD274) enhances HCV-specific T cell immunity in chronic hepatitis C infection.

The establishment of a chronic hepatitis C (CHC) infection is associated with defective HCV-specific T cell responses. Recent studies suggest that negative T cell regulators such as programmed death 1 (PD-1) contribute to the impairment of virus-specific T cell functions in chronic viral infections. However, the implication of peripheral monocytes from CHC patients in the inhibition of HCV-specific T cell responses is only partially defined. In this study, we found that B7-H1, a ligand of PD-1, was significantly up-regulated on monocytes of CHC patients. Proliferation of T cells in response to anti-CD3 antibody was directly suppressed by B7-H1+CD14+ monocytes, and this suppression was reversed by addition of antagonistic B7-H1 mAb. Furthermore, blocking of monocyte-associated B7-H1 (moB7-H1) significantly enhanced the frequency of IFN-gamma-producing, HCV-specific CD4+ and CD8+ effector T cells and the production of Th1 cytokines, such as IL-2 but not Th2 cytokines, including IL-4 and IL-10. Upon B7-H1 blockade, production of perforin was also increased in CD8+ T cells stimulated with HCV peptides. Our findings suggest that moB7-H1 inhibits HCV-specific CD4+ and CD8+ T lymphocyte proliferation and suppresses Th1 cytokine production and perforin secretion. Blockade of the B7-H1 pathway thus represents an attractive approach in the treatment of chronic HCV infection.

Characterization of the bone marrow adipocyte niche with three-dimensional electron microscopy.

Unlike white and brown adipose tissues, the bone marrow adipocyte (BMA) exists in a microenvironment containing unique populations of hematopoietic and skeletal cells. To study this microenvironment at the sub-cellular level, we performed a three-dimensional analysis of the ultrastructure of the BMA niche with focused ion beam scanning electron microscopy (FIB-SEM). This revealed that BMAs display hallmarks of metabolically active cells including polarized lipid deposits, a dense mitochondrial network, and areas of endoplasmic reticulum. The distinct orientations of the triacylglycerol droplets suggest that fatty acids are taken up and/or released in three key areas - at the endothelial interface, into the hematopoietic milieu, and at the bone surface. Near the sinusoidal vasculature, endothelial cells send finger-like projections into the surface of the BMA which terminate near regions of lipid within the BMA cytoplasm. In some regions, perivascular cells encase the BMA with their flattened cellular projections, limiting contacts with other cells in the niche. In the hematopoietic milieu, BMAT adipocytes of the proximal tibia interact extensively with maturing cells of the myeloid/granulocyte lineage. Associations with erythroblast islands are also prominent. At the bone surface, the BMA extends organelle and lipid-rich cytoplasmic regions toward areas of active osteoblasts. This suggests that the BMA may serve to partition nutrient utilization between diverse cellular compartments, serving as an energy-rich hub of the stromal-reticular network. Lastly, though immuno-EM, we've identified a subset of bone marrow adipocytes that are innervated by the sympathetic nervous system, providing an additional mechanism for regulation of the BMA. In summary, this work reveals that the bone marrow adipocyte is a dynamic cell with substantial capacity for interactions with the diverse components of its surrounding microenvironment. These local interactions likely contribute to its unique regulation relative to peripheral adipose tissues.

Gold-Nanocluster-Assisted Nanotransfer Printing Method for Metasurface Hologram Fabrication.

Given the development of nano/microscale patterning techniques, efforts are being made to use them for fabricating metasurfaces. In particular, by using abrupt phase discontinuities, it is possible to generate holographic images from two-dimensional nanoscale-patterned metasurfaces. However, the fabrication of metasurface holograms is hindered by the high costs and long fabrication time involved, because the process requires expensive equipment such as that for electron-beam lithography. Therefore, it is difficult to realize metasurface holograms in a fast and repetitive manner. In this study, we propose a method for fabricating metasurface holograms based on the nanotransfer printing of the desired nanoscale patterns, which is assisted by Au nanoclusters, while controlling the bonding energy based on the shape of the deposited Au layer. Robust covalent bonds are formed between the Si of the adhesive used and the O of the SiO2 layer in order to transfer the deposited Au onto the transparent substrate quickly. It was found that the fabricated metasurface hologram coincides with the one designed by computer-generated holography. The proposed method should lead to a significant breakthrough in the fabrication of holograms based on different types of metasurfaces at a low cost in a fast, repetitive manner with various metals.

Computer-Assisted Optimization of the Acetabular Rotation in Periacetabular Osteotomy Using Patient's Anatomy-Specific Finite Element Analysis.

Periacetabular osteotomy (PAO) is a complex surgical procedure to restore acetabular coverage in the dysplastic hip, and the amount of acetabular rotation during PAO plays a key role. Using computational simulations, this study assessed the optimal direction and amount of the acetabular rotation in three dimensions for a patient undergoing PAO. Anatomy-specific finite element (FE) models of the hip were constructed based on clinical CT images. The calculated acetabular rotation during PAO were 9.7°, 18°, and 4.3° in sagittal, coronal, and transverse planes, respectively. Based on the actual acetabular rotations, twelve postoperative FE models were generated. An optimal position was found by gradually varying the amount of the acetabular rotations in each anatomical plane. The coronal plane was found to be the principal rotational plane, which showed the strongest effects on joint contact pressure compared to other planes. It is suggested that rotation in the coronal plane of the osteotomized acetabulum is one of the primary surgical parameters to achieve the optimal clinical outcome for a given patient.

NecroX-5 prevents breast cancer metastasis by AKT inhibition via reducing intracellular calcium levels.

A major goal of breast cancer research is to prevent the molecular events that lead to tumour metastasis. It is well-established that both cytoplasmic and mitochondrial reactive oxygen species (ROS) play important roles in cell migration and metastasis. Accordingly, this study examined the molecular mechanisms of the anti-metastatic effects of NecroX-5, a mitochondrial ROS scavenger. NecroX-5 inhibited lung cancer metastasis by ameliorating migration in a mouse model. In human cancer cells, the inhibition of migration by NecroX-5 is cell type-dependent. We observed that the effect of NecroX-5 correlated with a reduction in mitochondrial ROS, but mitochondrial ROS reduction by MitoQ did not inhibit cell migration. NecroX-5 decreased intracellular calcium concentration by blocking Ca2+ influx, which mediated the inhibition of cell migration, AKT downregulation and the reduction of mitochondrial ROS levels. However, the reduction of mitochondrial ROS was not associated with supressed migration and AKT downregulation. Our study demonstrates the potential of NecroX-5 as an inhibitor of breast cancer metastasis.

Improvement of neutralizing activity of human scFv antibodies against hepatitis B virus binding using CDR3 V(H) mutant library.

CDR3 of the heavy-chain variable region of immunoglobulin is a region in which somatic mutation occurs heavily after secondary antibody response, resulting in an affinity maturation of antibodies in vivo. The aim of this study was to improve the affinity of a human single-chain variable fragment (scFv) specific for pre-S1 of hepatitis B virus (HBV) by introducing random mutagenesis in CDR3 variable region of heavy chain (V(H)) of the parental scFv clone 1E4. By using a BIAcore for panning and screening, we have selected three clones (A9, B2, and B9) with lower highest affinity (K(D)) than 1E4. Affinities of selected clones ranged from 1.7 x 10(7) mol/L to 6.3 x 10(8) mol/L, which were increased by factors of 1.4 to 4.0, respectively, compared to the parental clone. Binding inhibition assay using flow cytometry and polymerase chain reaction revealed that B2 (6.4 x 10(8) mol/L) had a higher neutralizing activity against pre-S1 or HBV virion binding to liver cell line. This anti-pre-S1 scFv can be considered as a potential therapeutic tool for a passive immunotherapy for HBV infection.

Early Postnatal NT-3 Gene Delivery Enhances Hearing Acquisition in the Developmental Period.

OBJECTIVES/HYPOTHESIS: The purpose of this study was to investigate the effect of early postnatal neurotrophin-3 (NT3) support on hearing acquisition. STUDY DESIGN: A prospective experimental animal study. METHODS: Adenoviral (Ad) vectors expressing green fluorescence protein (GFP) alone or in combination with NT3 were injected into the scala tympani through the round window of 5-postnatal-day-old (P5) rats. Changes in NT3 mRNA level, hearing thresholds, and morphological studies were done after the viral vector injection. RESULTS: NT3 mRNA was significantly increased in the Ad-GFP-NT3 group compared to the normal-developmental group and Ad-GFP alone group. GFP was widely expressed in the cochlea such as in the hair cells, supporting cell area, and spiral ganglion neurons. Auditory brainstem response thresholds were significantly lower in the Ad-GFP-NT3 group compared to the normal-developmental group and Ad-GFP alone group at P15. CONCLUSIONS: These results show that early postnatal NT3 overexpression may accelerate the acquisition of hearing in rats. LEVEL OF EVIDENCE: NA Laryngoscope, 126:E379-E385, 2016.