Spontaneous differentiation of human induced pluripotent stem cells to odorant-responsive olfactory sensory neurons.

Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells (iPSCs), can differentiate into almost all cell types and are anticipated to have significant applications in the field of regenerative medicine. However, there are no reports of successfully directing iPSCs to become functional olfactory sensory neurons (OSNs) capable of selectively receiving odorant compounds. In this study, we employed dual SMAD inhibition and fibroblast growth factor 8 (FGF-8, reported to dictate olfactory fates) along with N-2 and B-27 supplements in the culture medium to efficiently induce the differentiation of iPSCs into neuronal cells with olfactory function through olfactory placode. Temporal gene expression and expression of OSN-specific markers during differentiation indicated that the expression of olfactory marker proteins and various olfactory receptors (ORs), which are markers of mature OSNs, was observed after approximately one month of differentiation culture, irrespective of the differentiation cues, suggesting differentiation into OSNs. Cells that exhibited specific responses to odorant compounds were identified after administering odorant compounds to differentiated iPSC-derived OSNs. This suggests the spontaneous generation of functional OSNs expressing diverse ORs that respond to odorant compounds from iPSCs.

Suppression of lncRNA OIP5-AS1 Attenuates Apoptosis and Inflammation, and Promotes Proliferation by Mediating miR-25-3p Expression in Lipopolysaccharide-Induced Myocardial Injury.

Purpose: Long non-coding RNAs (LncRNAs) OIP5-AS1 and miR-25-3p play important roles in myocardial injury, whereas their roles in lipopolysaccharide (LPS)-induced myocardial injury remain unknown. The purpose of our study was to investigate the functional mechanisms of OIP5-AS1 and miR-25-3p in LPS-induced myocardial injury. Methods: Rats and H9C2 cells were treated with LPS to establish the model of myocardial injury in vivo and in vitro, respectively. The expression levels of OIP5-AS1 and miR-25-3p were determined by quantitative reverse transcriptase-polymerase chain reaction. Enzyme-linked immunosorbent assay was performed to measure the serum levels of IL-6 and TNF-α. The relationship between OIP5-AS1 and miR-25-3p/NOX4 was determined by luciferase reporter assay and/or RNA immunoprecipitation assay. The apoptosis rate was detected by flow cytometry, and cell viability was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Western blot was performed to detect the protein levels of Bax, Bcl-2, caspase3, c-caspase3, NOX4, and p-NF-κB p65/NF-κB p65. Results: OIP5-AS1 was up-regulated, and miR-25-3p was down-regulated in myocardial tissues of LPS-induced rats and LPS-treated H9C2 cells. Knockdown of OIP5-AS1 relieved the myocardial injury in LPS-induced rats. Knockdown of OIP5-AS1 also inhibited the inflammation and apoptosis of myocardial cells in vivo, which was subsequently confirmed by in vitro experiments. In addition, OIP5-AS1 targeted miR-25-3p. MiR-25-3p mimics reversed the effects of OIP5-AS1 overexpression on promoting cell apoptosis and inflammation and on inhibiting cell viability. Besides, miR-25-3p mimics blocked the NOX4/NF-κB signalling pathway in LPS-induced H9C2 cells. Conclusion: Silencing of lncRNA OIP5-AS1 alleviated LPS-induced myocardial injury by regulating miR-25-3p.

Ultra-high-resolution quanta image sensor with reliable photon-number-resolving and high dynamic range capabilities.

Superior low-light and high dynamic range (HDR) imaging performance with ultra-high pixel resolution are widely sought after in the imaging world. The quanta image sensor (QIS) concept was proposed in 2005 as the next paradigm in solid-state image sensors after charge coupled devices (CCD) and complementary metal oxide semiconductor (CMOS) active pixel sensors. This next-generation image sensor would contain hundreds of millions to billions of small pixels with photon-number-resolving and HDR capabilities, providing superior imaging performance over CCD and conventional CMOS sensors. In this article, we present a 163 megapixel QIS that enables both reliable photon-number-resolving and high dynamic range imaging in a single device. This is the highest pixel resolution ever reported among low-noise image sensors with photon-number-resolving capability. This QIS was fabricated with a standard, state-of-the-art CMOS process with 2-layer wafer stacking and backside illumination. Reliable photon-number-resolving is demonstrated with an average read noise of 0.35 e- rms at room temperature operation, enabling industry leading low-light imaging performance. Additionally, a dynamic range of 95 dB is realized due to the extremely low noise floor and an extended full-well capacity of 20k e-. The design, operating principles, experimental results, and imaging performance of this QIS device are discussed.

Threshold Uniformity Improvement in 1b Quanta Image Sensor Readout Circuit.

A new readout architecture for single-bit quanta image sensor (QIS) consisting of a capacitive transimpedance amplifier (CTIA) before a 1-bit quantizer to improve the threshold uniformity of the readout cluster is proposed in this paper. The 1-bit quantizer in the previous single-bit QIS had significant threshold non-uniformity likely caused by the fluctuation of the common-mode voltage of the jot output. To guarantee the stability of the common-mode voltage of input signals fed to the 1-bit quantizer, the CTIA is added before the 1-bit quantizer. A pipeline operation mode is also proposed so the CTIA and 1-bit ADC can work at the same time, thereby reducing the CTIA power consumption. A 2048 × 1024 high-speed test chip was implemented with 45 nm/65 nm stacked backside illuminated (BSI) CMOS image sensor (CIS) process and tested. According to the measured D-log-H results, a good threshold uniformity in the range of 0.3 to 0.8 e- for all readout clusters is demonstrated at 500 frame per second (fps) equivalent timing with 68 mW power consumption.

Effect of tyrosinase and caffeic acid crosslinking of turbot parvalbumin on the digestibility, and release of mediators and cytokines from activated RBL-2H3 cells.

Turbot can induce allergy in susceptible individuals due to the presence of parvalbumin (PV), a major fish allergen. This study aimed at evaluating the digestibility and the ability of PV to elicit the release of cellular degranulation, following treatment with tyrosinase (PV-Tyr), caffeic acid (PV-CA) and in combination (PV-Tyr/CA), using in vitro digestion and RBL-2H3 (passive rat basophil leukemia) cell line. The digestion assay products revealed that the stability of PV in simulated gastric fluid (SGF) was stronger, while in simulated intestinal fluid (SIF) was rather weak. Western blot analysis revealed that the IgG-binding abilities of the cross-linked PV were markedly reduced. Moreover, crosslinking hampered the release of cellular degranulation process in RBL-2H3 cell lines. PV-Tyr/CA showed highly significant reduction in the release rate of ß-hexosaminidase (66.02%), histamine (35.01%), tryptase (29.25%), cysteinyl leukotrienes (29.72%), prostaglandin D2 (34.96%), IL-4 (43.99%) and IL-13 (38.93%) and shown potential in developing hypoallergenic fish products.

Megapixel photon-counting color imaging using quanta image sensor.

Quanta Image Sensor (QIS) is a single-photon detector designed for extremely low light imaging conditions. Majority of the existing QIS prototypes are monochrome based on single-photon avalanche diodes (SPAD). Passive color imaging has not been demonstrated with single-photon detectors due to the intrinsic difficulty of shrinking the pixel size and increasing the spatial resolution while maintaining acceptable intra-pixel cross-talk. In this paper, we present image reconstruction of the first color QIS with a resolution of 1024 × 1024 pixels, supporting both single-bit and multi-bit photon counting capability. Our color image reconstruction is enabled by a customized joint demosaicing-denoising algorithm, leveraging truncated Poisson statistics andvariance stabilizing transforms. Experimental results of the new sensor and algorithm demonstrate superior color imaging performance for very low-light conditions with a mean exposure of as low as a few photons per pixel in both real and simulated images.

Prevalence and distribution of HPV types in genital warts in Xi'an, China: a prospective study.

OBJECTIVES: To characterise the prevalence and distribution of human papillomavirus (HPV) types in genital warts in Xi'an, China. METHODS: This prospective study was conducted in Shaanxi Provincial Institute for Skin Disease and STD Control (SPISSC) between September 2014 and April 2017. Genital wart samples were obtained from 879 patients, including 512 men and 367 women. HPV genotyping was performed by using an automatic nucleic acid hybridisation system. RESULTS: Of the 879 patients with genital warts, the detectable rates of low-risk, high-risk and total HPV types were 45.4%, 34.5% and 57.8%, respectively. The detectable rate of low-risk HPV types (45.4%) was significantly higher than that of high-risk HPV types (34.5%) (χ2=21.85, p<0.01). The detectable rate of low-risk HPV types of men (52.3%) was significantly higher than that of women (35.7%) (χ2=23.90, p<0.01). The detectable rates of one HPV type infection and two and three or more HPV type coinfections were 26.1%, 17.5% and 14.2%, respectively. HPV6 (24.9%), HPV11 (17.9%), HPV52 (9.9%) and HPV16 (7.3%) were the four most common HPV types. CONCLUSIONS: The results of this study suggest that low-risk HPV types are major pathogens of genital warts, but high-risk HPV type infections and multiple HPV type coinfections are also common in genital warts. HPV6, 11, 52 and 16 are the four most common HPV types in genital wart in Xi'an, China.

Effect of tyrosinase-catalyzed crosslinking on the structure and allergenicity of turbot parvalbumin mediated by caffeic acid.

BACKGROUND: Enzymatic treatment of allergenic protein can alter their functional properties under a mild reaction condition due to specificity of enzymes. Phenolic compounds act as mediators and enhance the crosslinking reactions. The study aimed to assess the changes in the structure and immunoglobulin G (IgG) binding capacity of turbot parvalbumin (PV) upon crosslinking with tyrosinase (Tyr) in the absence and presence of caffeic acid. RESULTS: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the appearance of higher molecular weight bands (24, 36 kDa) in the crosslinked PV. The secondary structure of crosslinked PV became loosened and disordered. The results of intrinsic fluorescence and ultraviolet absorption spectral analyses, as well as surface hydrophobicity and free amino group analyses also revealed structural changes. As observed by western blot analysis, the intensity of the PV bands reduced upon Tyr treatment, indicating reduced binding of specific IgG to PV. Moreover, the indirect ELISA (enzyme-linked immunosorbent assay) analysis confirmed that the IgG binding ability of crosslinked PV was reduced 34.94%. CONCLUSION: Enzymatic treatment mitigated the allergenicity of fish PV, which was closely related to the alterations in the conformational structure. This treatment showed potential for developing hypoallergenic fish products under mild reaction conditions. © 2019 Society of Chemical Industry.

Impacts of glycation and transglutaminase-catalyzed glycosylation with glucosamine on the conformational structure and allergenicity of bovine ß-lactoglobulin.

ß-Lactoglobulin (ß-LG) is recognized as the major milk allergen. In this study, the effects of transglutaminase (TGase) and glucosamine (GlcN)-catalyzed glycosylation and glycation on the conformational structure and allergenicity of ß-LG were investigated. The formations of cross-linked peptides were demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and GlcN-conjugated modification was identified using matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Structural analysis revealed that glycosylation and glycation of ß-LG induced unfolding of the primary protein structure followed by a loss of the secondary structure. As revealed by circular dichroism (CD) spectroscopy, glycosylated ß-LG exhibited the highest increase in the ß-sheets from 32.6% to 40.4% (25 °C) and 44.2% (37 °C), and the percentage of α-helices decreased from 17.7% to 14.4% (25 °C) and 12.3% (37 °C), respectively. The tertiary and quaternary structures of ß-LG also changed significantly during glycosylation and glycation, along with reduced free amino groups and variation in surface hydrophobicity. Immunoblotting and indirect enzyme-linked immuno sorbent assay (ELISA) analyses demonstrated that the lowest IgG- and IgE-binding capacities of ß-LG were obtained following glycosylation at 37 °C, which were 52.7% and 56.3% lower than that of the native protein, respectively. The reduction in the antigenicity and potential allergenicity of glycosylated ß-LG was more pronounced compared to TGase treated- and glycated ß-LG, which correlated well with the structural changes. These results suggest that TGase-catalyzed glycosylation has more potential compared to glycation for mitigating the allergenic potential of milk products.

Effect of tyrosinase-aided crosslinking on the IgE binding potential and conformational structure of shrimp (Metapenaeus ensis) tropomyosin.

The present study was performed to determine crosslinking and oxidative reactions catalyzed by tyrosinase (Tyr), caffeic acid (CA) and their combination with respect to IgE binding potential and conformational structure of shrimp tropomyosin (TM). Cross-links and IgE binding potentials were analyzed by SDS-PAGE, western blot and indirect ELISA. While structural changes were characterized using surface hydrophobicity, ultraviolet (UV), fluorescence and circular dichroism (CD) spectroscopies. Maximum reduction in the IgG (37.19%) and IgE binding potentials (49.41%) were observed when treated with 2000 nkat/g Tyr + CA, as indicated by ELISA analyses. These findings correlated well with the denaturation of protein, as evident by slight blue shift and alterations in the ellipticities observed via structural analyses. The results demonstrated that addition of CA mediator with Tyr pronouncedly enhanced crosslinking, and altered the conformational structure, thereby mitigated allergenicity of TM, thus showing promise in developing novel food structures with reduced allergenic potential.

Mixture prepared with Chinese medicines in terms of releasing exterior cold and clearing interior heat: its efficacy and safety in patients with seasonal influenza-a randomized controlled trial.

OBJECTIVE: To assess the efficacy and safety in 105 patients with seasonal Influenza in Beijing, the mixture prepared with Chinese medicines follows the treatment regimen of releasing exterior cold and clearing interior heat. METHODS: Total 330 patients with seasonal influenza were enrolled and randomly and averagely divided into the Chinese herbal medicine, the western medicine and the Chinese patent medicine group. They were treated with Chinese medicine Oseltamivir Phosphate Capsules and the Scattering Wind and Resolving Toxins Capsules. The main efficacy indicators were the antifebrile onset time and recovery time of body temperature. The efficacy and safety of the mixture was scientifically evaluated. Comparisons of several variables were analyzed. RESULTS: Median antifebrile onset time of the Chinese herbal medicine group was significantly shorter than the western medicine group (P < 0.05) and the Chinese patent medicine group (P < 0.05). The median antifebrile recovery time of the Chinese herbal medicine group was significantly shorter than the Chinese patent medicine group (P < 0.05). The groups evaluated by TCM symptom pattern effect, both the Chinese herbal medicine group and Western Medicine group were better than the Chinese patent medicine group (P < 0.05). The disappearance rate of main symptoms and some minor symptom patterns of the Chinese herbal medicine group were higher than the other 2 groups. CONCLUSION: The mixture of releasing exterior cold and clearing interior heat could significantly shorten the fever time with safety.

The Quanta Image Sensor: Every Photon Counts.

The Quanta Image Sensor (QIS) was conceived when contemplating shrinking pixel sizes and storage capacities, and the steady increase in digital processing power. In the single-bit QIS, the output of each field is a binary bit plane, where each bit represents the presence or absence of at least one photoelectron in a photodetector. A series of bit planes is generated through high-speed readout, and a kernel or "cubicle" of bits (x, y, t) is used to create a single output image pixel. The size of the cubicle can be adjusted post-acquisition to optimize image quality. The specialized sub-diffraction-limit photodetectors in the QIS are referred to as "jots" and a QIS may have a gigajot or more, read out at 1000 fps, for a data rate exceeding 1 Tb/s. Basically, we are trying to count photons as they arrive at the sensor. This paper reviews the QIS concept and its imaging characteristics. Recent progress towards realizing the QIS for commercial and scientific purposes is discussed. This includes implementation of a pump-gate jot device in a 65 nm CIS BSI process yielding read noise as low as 0.22 e- r.m.s. and conversion gain as high as 420 µV/e-, power efficient readout electronics, currently as low as 0.4 pJ/b in the same process, creating high dynamic range images from jot data, and understanding the imaging characteristics of single-bit and multi-bit QIS devices. The QIS represents a possible major paradigm shift in image capture.

Quantum Random Number Generation Using a Quanta Image Sensor.

A new quantum random number generation method is proposed. The method is based on the randomness of the photon emission process and the single photon counting capability of the Quanta Image Sensor (QIS). It has the potential to generate high-quality random numbers with remarkable data output rate. In this paper, the principle of photon statistics and theory of entropy are discussed. Sample data were collected with QIS jot device, and its randomness quality was analyzed. The randomness assessment method and results are discussed.

Effect of 4-hydroxy-2-nonenal treatment on the IgE binding capacity and structure of shrimp (Metapenaeus ensis) tropomyosin.

Lipid peroxidation can react with free amines of proteins and induce modification of structural and functional properties. This study presents the IgE binding capacity and structural changes of shrimp tropomyosin (TM) under oxidative stress with 4-hydroxy-2-nonenal (HNE). IgE binding capacity was evaluated with the dot-blot assay and inhibition enzyme-linked immunosorbent assay. A decrease in IgE binding capacity of TM was found with 0.01mM HNE treatment, which was more significant when the HNE concentration was increased to 0.5mM. The conformational changes of TM, as characterized by fluorescence spectroscopy, circular dichroism spectroscopy and ultraviolet absorption spectroscopy, correlated well with IgE binding capacity changes. Further LC-ESI-MS/MS analyses showed that the side-chain groups of alanine, leucine, lysine and histidine had been modified by HNE. These results suggested that the HNE-induced conformational changes of TM significantly influenced its allergenicity and that these changes were caused by the modification of specific amino acids residues.

Reduction of CMOS Image Sensor Read Noise to Enable Photon Counting.

Recent activity in photon counting CMOS image sensors (CIS) has been directed to reduction of read noise. Many approaches and methods have been reported. This work is focused on providing sub 1 e(-) read noise by design and operation of the binary and small signal readout of photon counting CIS. Compensation of transfer gate feed-through was used to provide substantially reduced CDS time and source follower (SF) bandwidth. SF read noise was reduced by a factor of 3 with this method. This method can be applied broadly to CIS devices to reduce the read noise for small signals to enable use as a photon counting sensor.

Exploring association between gastrointestinal heat retention syndrome and recurrent respiratory tract infections in children: a prospective cohort study.

BACKGROUND: Recurrent respiratory tract infections (RRTIs) have a negative impact on both children's health and family wellbeing. Deficiency of ZhengQi used to be an instinct factor driving RRTI in Traditional Chinese Medicine (TCM). Our clinical observations suggest that children with gastrointestinal heat retention syndrome (GHRS) may have a greater risk of catching respiratory tract infections (RTIs). GHRS is a new predisposing factor for RRTI and it is dietary related. This study is aimed to explore association between GHRS and RRTI. METHODS: A prospective cohort study has been conducted in Beijing, China; children aged 1-18 were enrolled. TCM symptoms, demographic and physiological characteristics were recorded by using semi-structured questionnaire. GHRS was considered as a predisposing factor. Children were followed up for next 12 months. We contacted with their parents using a face-to-face questionnaire survey, via email or phone every 3 months. Episodes of RTIs were recorded in detail. RESULTS: Three hundred thirty four children were enrolled and 307 (91.92%) followed up for 12 months. The incidence of RTI was 4.32 episodes per child-year (95 % CI 4.03-4.61). 69 (43.13%) children in the group with GHRS suffered from RRTI; there were 48 (32.65%) children in group without GHRS. The risk ratio (RR) value of RRTI occurrence was 1.32 (95 % CI 0.91-1.91, P = 0.139), and the attributable risk percent (AR%) was 24.28%. Dry stool and irritability were positively correlated with RTI episodes, age and BMI were negatively correlated with RTI episodes in a linear regression model. Dry stool (OR = 1.510) was positively correlated with RRTI occurrence, age (OR = 0.889) and BMI (OR = 0.858) were negatively correlated with RRTI occurrence in our logistic regression model. CONCLUSIONS: GHRS is associated with RRTI in this cohort. Dry stool was positively associated with RRTI, and BMI was negatively associated with RRTI. Studies with larger sample size and longer follow up are needed to further evaluate this association. Relieving GHRS should be considered when TCM practitioners treat RRTI children, and this may protect children from suffering RTIs. TRIAL REGISTRATION: Chinese Clinical Trial Registry Number: ChiCTR-CCH-13003756.