16 research outputs found
The effect of Dnaaf5 gene dosage on primary ciliary dyskinesia phenotypes
DNAAF5 is a dynein motor assembly factor associated with the autosomal heterogenic recessive condition of motile cilia, primary ciliary dyskinesia (PCD). The effects of allele heterozygosity on motile cilia function are unknown. We used CRISPR-Cas9 genome editing in mice to recreate a human missense variant identified in patients with mild PCD and a second, frameshift-null deletion in Dnaaf5. Litters with Dnaaf5 heteroallelic variants showed distinct missense and null gene dosage effects. Homozygosity for the null Dnaaf5 alleles was embryonic lethal. Compound heterozygous animals with the missense and null alleles showed severe disease manifesting as hydrocephalus and early lethality. However, animals homozygous for the missense mutation had improved survival, with partially preserved cilia function and motor assembly observed by ultrastructure analysis. Notably, the same variant alleles exhibited divergent cilia function across different multiciliated tissues. Proteomic analysis of isolated airway cilia from mutant mice revealed reduction in some axonemal regulatory and structural proteins not previously reported in DNAAF5 variants. Transcriptional analysis of mouse and human mutant cells showed increased expression of genes coding for axonemal proteins. These findings suggest allele-specific and tissue-specific molecular requirements for cilia motor assembly that may affect disease phenotypes and clinical trajectory in motile ciliopathies
Purification, Characterization, and Mode of Action of Pentocin JL-1, a Novel Bacteriocin Isolated from Lactobacillus pentosus, against Drug-Resistant Staphylococcus aureus
Staphylococcus aureus and its drug-resistant strains, which threaten public health and food safety, are in need of effective control by biopreservatives. A novel bacteriocin, pentocin JL-1, produced by Lactobacillus pentosus that was isolated from the intestinal tract of Chiloscyllium punctatum, was purified by a four-step chromatographic process. Mass spectrometry based on MALDI-TOF indicated that pentocin JL-1 has a molecular mass of 2987.23 Da. Only six of the twenty-five amino acids could be identified by Edman degradation. This bacteriocin is thermostable and tolerates a pH range of 5–7. Also, it is sensitive to proteinase K, trypsin, pepsin, and alkaline protease. This bacteriocin has a broad inhibitory spectrum against both Gram-positive and Gram-negative strains and in particular is effective against multidrug-resistant S. aureus. Additionally, we showed that the cell membrane is the target of pentocin JL-1 against methicillin-resistant S. aureus (MRSA), causing a loss of proton motive force. Furthermore, pentocin JL-1 has a drastic impact on the structure and integrity of MRSA cells. These results suggest that pentocin JL-1 has potential as a biopreservative in the food industry
Polymer blend lithography: A versatile method to fabricate nanopatterned self-assembled monolayers
A rapid and cost-effective lithographic method, polymer blend lithography (PBL), is reported to produce patterned self-assembled monolayers (SAM) on solid substrates featuring two or three different chemical functionalities. For the pattern generation we use the phase separation of two immiscible polymers in a blend solution during a spin-coating process. By controlling the spin-coating parameters and conditions, including the ambient atmosphere (humidity), the molar mass of the polystyrene (PS) and poly(methyl methacrylate) (PMMA), and the mass ratio between the two polymers in the blend solution, the formation of a purely lateral morphology (PS islands standing on the substrate while isolated in the PMMA matrix) can be reproducibly induced. Either of the formed phases (PS or PMMA) can be selectively dissolved afterwards, and the remaining phase can be used as a lift-off mask for the formation of a nanopatterned functional silane monolayer. This “monolayer copy” of the polymer phase morphology has a topographic contrast of about 1.3 nm. A demonstration of tuning of the PS island diameter is given by changing the molar mass of PS. Moreover, polymer blend lithography can provide the possibility of fabricating a surface with three different chemical components: This is demonstrated by inducing breath figures (evaporated condensed entity) at higher humidity during the spin-coating process. Here we demonstrate the formation of a lateral pattern consisting of regions covered with 1H,1H,2H,2H-perfluorodecyltrichlorosilane (FDTS) and (3-aminopropyl)triethoxysilane (APTES), and at the same time featuring regions of bare SiOx. The patterning process could be applied even on meter-sized substrates with various functional SAM molecules, making this process suitable for the rapid preparation of quasi two-dimensional nanopatterned functional substrates, e.g., for the template-controlled growth of ZnO nanostructures
Development of Simplified Models for Non-Destructive Hyperspectral Imaging Monitoring of S-ovalbumin Content in Eggs during Storage
S-ovalbumin content is an indicator of egg freshness and has an important impact on the quality of processed foods. The objective of this study is to develop simplified models for monitoring the S-ovalbumin content of eggs during storage using hyperspectral imaging (HSI) and multivariate analysis. The hyperspectral images of egg samples at different storage periods were collected in the wavelength range of 401–1002 nm, and the reference S-ovalbumin content was determined by spectrophotometry. The standard normal variate (SNV) was employed to preprocess the raw spectral data. To simplify the calibration models, competitive adaptive reweighted sampling (CARS) was applied to select feature wavelengths from the whole spectral range. Based on the full and feature wavelengths, partial least squares regression (PLSR) and least squares support vector machine (LSSVM) models were developed, in which the simplified LSSVM model yielded the best performance with a coefficient of determination for prediction (R2P) of 0.918 and a root mean square error for prediction (RMSEP) of 7.215%. By transferring the quantitative model to the pixels of hyperspectral images, the visualizing distribution maps were generated, providing an intuitive and comprehensive evaluation for the S-ovalbumin content of eggs, which helps to understand the conversion of ovalbumin into S-ovalbumin during storage. The results provided the possibility of implementing a multispectral imaging technique for online monitoring the S-ovalbumin content of eggs
Quantitation by Liquid Chromatography–Nanoelectrospray Ionization–High-Resolution Tandem Mass Spectrometry of Multiple DNA Adducts Related to Cigarette Smoking in Oral Cells in the Shanghai Cohort Study
We developed a liquid chromatography–nanoelectrospray
ionization–high-resolution
tandem mass spectrometry (LC–NSI–HRMS/MS) method for
simultaneous quantitative analysis of 5 oral cell DNA adducts associated
with cigarette smoking: (8R/S)-3-(2′-deoxyribos-1′-yl)-5,6,7,8-tetrahydro-8-hydroxypyrimido[1,2-a]purine-10(3H)-one (γ-OH-Acr-dGuo, 1) from acrolein; (6S,8S and 6R,8R)-3-(2′-deoxyribos-1′-yl)-5,6,7,8-tetrahydro-8-hydroxy-6-methylpyrimido[1,2-a]purine-10(3H)-one [(6S,8S)γ-OH-Cro-dGuo, 2; and (6R,8R)γ-OH-Cro-dGuo, 3] from crotonaldehyde; 1,N6-etheno-dAdo (4) from acrylonitrile, vinyl chloride,
lipid peroxidation, and inflammation; and 8-oxo-dGuo (5) from oxidative damage. Oral cell DNA was isolated in the presence
of glutathione to prevent artifact formation. Clear LC–NSI–HRMS/MS
chromatograms were obtained allowing quantitation of each adduct using
the appropriately labeled internal standards. The accuracy and precision
of the method were validated, and the assay limit of quantitation
was 5 fmol/μmol dGuo for adducts 1–4 and 20 fmol/μmol for adduct 5. The assay
was applied to 80 buccal cell samples selected from those collected
in the Shanghai Cohort Study: 40 from current smokers and 40 from
never smokers. Significant differences were found in all adduct levels
between smokers and nonsmokers. Levels of 8-oxo-dGuo (5) were at least 3000 times greater than those of the other adducts
in both smokers and nonsmokers, and the difference between amounts
of this adduct in smokers versus nonsmokers, while significant (P = 0.013), was not as great as the differences of the other
DNA adducts between smokers and nonsmokers (P-values
all less than 0.001). No significant relationship of adduct levels
to risk of lung cancer incidence was found. This study provides a
new LC–NSI–HRMS/MS methodology for the quantitation
of diverse DNA adducts resulting from exposure to the α,β-unsaturated
aldehydes acrolein and crotonaldehyde, inflammation, and oxidative
damage which are all associated with carcinogenesis. We anticipate
application of this assay in ongoing studies of the molecular epidemiology
of cancers of the lung and oral cavity related to cigarette smoking
Occurrence and risk of iodinated X-ray contrast media in source and tap water from Jiangsu province, China
Microcontaminants in the water environment have received increasing attention due to their adverse effects on human health and wildlife. However, iodinated X-ray contrast media (ICM), a type of microcontaminants, have not yet been systematically documented in source and tap water. This study investigated ICM in water samples via a sampling activity from 25 drinking water sources and their corresponding 30 household taps in south-central Jiangsu Province, China. The total concentrations of ICM ranged from 14.2 to 138.5 ng/L in source water and 3.7 to 101.3 ng/L in tap water, respectively. The calculated average water treatment efficiency to remove ICM is 38.3% with large variation under different processes (ranging from 7.3% to 75.7%), which implied that ICM could not be effectively removed using current treatment technologies. By integrating other ICM into the predominant compound iohexol with relative potency factors, the health risks of total ICM through water consumption were assessed using the Monte Carlo simulation. The results concluded that the risk of ingesting ICM through tap water was not a major health concern for adults, teens, or children in the study area. Nevertheless, due to the lack of long-term toxicity data relevant for humans for ICM, this risk may be underestimated, which requires further research
Effects of stabilization and sludge properties in a combined process of anaerobic digestion and thermophilic aerobic digestion
Security Scheme Enhancement for Voice over Wireless Networks
This paper proposes a lightweight and low energy encryption algorithm to secure voice traffic over wireless networks. The proposed algorithm meets the requirements of voice traffic and is suitable for wireless devices. It is capable to reduce the execution time and power consumption of the encryption process compared with the state of the art standard algorithm and at the same time maintains the desired security (confidentiality) level. The proposed algorithm employs similar methods with those used in the Advanced Encryption Standard algorithm (AES), with enhancements considering the limitations of wireless devices. A range of simulation scenarios are developed to test the validity of the proposed algorithm in reducing delay and energy required for the encryption process, besides, not compromising the security level. The results show significant improvements in the network and security performance metrics. A comparison between the proposed algorithm, the AES and other lightweight algorithms proposed by other research works is conducted and the results are promising. Using the proposed algorithm, a significant amount of time and energy consumption reduction is achieved to reach approximately 35% enhancement over the standard AES algorithm accompanied with good-level of complexity in the encryption process, making it more suitable for the wireless environment