A Homeodomain-Containing Transcriptional Factor PoHtf1 Regulated the Development and Cellulase Expression in Penicillium oxalicum

Homeodomain-containing transcription factors (Htfs) play important roles in animals, fungi, and plants during some developmental processes. Here, a homeodomain-containing transcription factor PoHtf1 was functionally characterized in the cellulase-producing fungi Penicillium oxalicum 114-2. PoHtf1 was shown to participate in colony growth and conidiation through regulating the expression of its downstream transcription factor BrlA, the key regulator of conidiation in P. oxalicum 114-2. Additionally, PoHtf1 inhibited the expression of the major cellulase genes by coordinated regulation of cellulolytic regulators CreA, AmyR, ClrB, and XlnR. Furthermore, transcriptome analysis showed that PoHtf1 participated in the secondary metabolism including the pathway synthesizing conidial yellow pigment. These data show that PoHtf1 mediates the complex transcriptional-regulatory network cascade between developmental processes and cellulolytic gene expression in P. oxalicum 114-2. Our results should assist the development of strategies for the metabolic engineering of mutants for applications in the enzymatic hydrolysis for biochemical production

Sulforaphane Inhibits Foam Cell Formation and Atherosclerosis via Mechanisms Involving the Modulation of Macrophage Cholesterol Transport and the Related Phenotype

Sulforaphane (SFN), an isothiocyanate, is one of the major dietary phytochemicals found in cruciferous vegetables. Many studies suggest that SFN can protect against cancer and cardiometabolic diseases. Despite the proposed systemic and local vascular protective mecha-nisms, SFN’s potential to inhibit atherogenesis by targeting macrophages remains unknown. In this study, in high-fat-diet-fed ApoE-deficient (ApoE-/-) mice, oral SFN treatment improved dyslipidemia and inhibited atherosclerotic plaque formation and the unstable phenotype, as demonstrated by reductions in the lesion areas in both the aortic sinus and whole aorta, per-centages of necrotic cores, vascular macrophage infiltration and reactive oxygen species (ROS) generation. In THP-1-derived macrophages, SFN pre-administration alleviated oxidized low-density lipoprotein (ox-LDL)-induced lipid accumulation, oxidative stress and mitochondrial injury. Moreover, a functional study revealed that peritoneal macrophages isolated from SFN-treated mice exhibited attenuated cholesterol influx and enhanced apolipoprotein A-I (apoA-I)- and high-density lipoprotein (HDL)-mediated cholesterol efflux. Mechanistic analysis revealed that SFN supplementation induced both intralesional and intraperitoneal macrophage phenotypic switching toward high expression of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and ATP binding cassette subfamily A/G member 1 (ABCA1/G1) and low expression of peroxisome proliferator-activated receptor γ (PPARγ) and cluster of differen-tiation 36 (CD36), which was further validated by the aortic protein expression. These results suggest that the regulation of macrophages cholesterol transport and accumulation may be mainly responsible for SFN's potential atheroprotective properties, and the regulatory mecha-nisms might involve upregulating ABCA1/G1 and downregulating CD36 via the modulation of PPARγ and Nrf2

Molecular characterization of florfenicol and oxazolidinone resistance in Enterococcus isolates from animals in China

Florfenicol is widely used for the treatment of bacterial infections in domestic animals. The aim of this study was to analyze the molecular mechanisms of florfenicol and oxazolidinone resistance in Enterococcus isolates from anal feces of domestic animals. The minimum inhibitory concentration (MIC) levels were determined by the agar dilution method. Polymerase chain reaction (PCR) was performed to analyze the distribution of the resistance genes. Whole-genome sequencing and comparative plasmid analysis was conducted to analyze the resistance gene environment. A total of 351 non-duplicated enteric strains were obtained. Among these isolates, 22 Enterococcus isolates, including 19 Enterococcus. faecium and 3 Enterococcus. faecalis, were further studied. 31 florfenicol resistance genes (13 fexA, 3 fexB, 12 optrA, and 3 poxtA genes) were identified in 15 of the 19 E. faecium isolates, and no florfenicol or oxazolidinone resistance genes were identified in 3 E. faecalis isolates. Whole-genome sequencing of E. faecium P47, which had all four florfenicol and oxazolidinone resistance genes and high MIC levels for both florfenicol (256 mg/L) and linezolid (8 mg/L), revealed that it contained a chromosome and 3 plasmids (pP47-27, pP47-61, and pP47-180). The four florfenicol and oxazolidinone resistance genes were all related to the insertion sequences IS1216 and located on two smaller plasmids. The genes fexB and poxtA encoded in pP47-27, while fexA and optrA encoded in the conjugative plasmid pP47-61. Comparative analysis of homologous plasmids revealed that the sequences with high identities were plasmid sequences from various Enterococcus species except for the Tn6349 sequence from a Staphylococcus aureus chromosome (MH746818.1). The current study revealed that florfenicol and oxazolidinone resistance genes (fexA, fexB, poxtA, and optrA) were widely distributed in Enterococcus isolates from animal in China. The mobile genetic elements, including the insertion sequences and conjugative plasmid, played an important role in the horizontal transfer of florfenicol and oxazolidinone resistance

Harnessing CRISPR-Cas12a and Nanotechnology for the Rapid and Accurate Detection of DNA Viruses

In recent years, the world has faced numerous pandemics caused by viral diseases, leading to considerable morbidity, mortality, and economic disruptions. Developing vaccines for pandemics can be a challenging task due to the rapid mutation of viruses and the significant investment of time required for their development. While real-time polymerase chain reaction and immunoassay techniques are frequently utilized for viral diagnostics, they possess inherent limitations. Therefore, there is a pressing need to rapidly and accurately identify viral infections to manage diseases effectively. This work explores the use of nanotechnology and clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) proteins to achieve rapid and accurate viral identification. One biosensing assay and two microdevices are presented, each with innovative features. The biosensing assay employs Quantum dots as indicators and Cas-mediated nucleic acid probe cleavage to provide a simple “Yes-or-No” readout. By adding a magnetic bead washing step, the assay delivers comparable detection sensitivity in both buffer and plasma, largely eliminating high background issues raised in other publications. One of the microdevices presented couples the isothermal amplification technique with the CRISPR-Cas assay, achieving ultra-sensitivity with the help of a hand warmer pouch for heating. This microdevice is pre-loaded with reagents and can be operated by simply pulling rods, making it possible for ordinary people to perform on-site diagnostics. The other microdevice incorporates high-aspect-ratio micropillars and the work demonstrates that using high-density micropillars can achieve higher detection sensitivity

A Novel Flow Sensor for a Smart Shunt System

Cerebrospinal fluid (CSF) is a body fluid contained in the brain ventricles and the cranial and spinal subarachnoid spaces. It plays an essential role in regulating neuronal function and maintaining homeostasis of interstitial fluid in the brain. Hydrocephalus is a neurological condition classified by the abnormal accumulation of cerebrospinal fluid (CSF) in the brain. It occurs due to a mismatch between CSF production and its absorption within the brain. The current preferred treatment for hydrocephalus is to post a CSF shunt system by medical procedure. However, shunt failure is relatively common among patients due to shunt malfunction, obstruction and infection. This thesis proposes a microdevice to determine the shunt performance by measuring the flow rate in the compartment. The concept behind this microdevice is to establish a feedback system of measured conditions and analyzed data, providing parameters to examine shunt efficiency. The purpose of this microdevice is to improve current hydrocephalus treatment, assist neurologists to evaluate shunt performance, and reduce the chance of missed diagnosis of shunt failure. The microdevice operates based on the principle of bending cantilever brought by the pressure of the fluid in the shunt tube. The laser light was reflected by the beam inside the compartment, and then captured by a microscopic camera. The changes in light intensity revealed the flow rates in the shunt tube. In this study, we have designed and fabricated this novel sensor, and demonstrated its normal operation between 500 ml/hr and 900 ml/hr. The results also showed that the sensor can endure long-term pressure of the fluid without permeant deformation

An Improved Innovation Robust Outliers Detection Method for Airborne Array Position and Orientation Measurement System

The airborne array position and orientation measurement system (array POS) is a key device for high-resolution multi-dimensional real-time imaging motion compensation of military reconnaissance mapping. Abnormal values will appear in array POS inertial devices and measurement data in an environment of strong interference, which often leads to a decrease or even divergence in the combination accuracy. The existing detection methods based on innovation characteristics are only sensitive to measurement outliers, which are the abnormal data caused by the strong interference environment. In this paper, an improved innovation robust outliers detection method is proposed, which is valid for both measurement outliers and inertial device outliers. First, the improved outliers detection method based on the innovation of array POS is described. The gain matrix is adaptively adjusted by using the statistical characteristics of innovation. At the same time, the information distribution coefficient is adaptively adjusted by using the filtering performance of the sub filter, which realizes the detection and correction of measurement outliers. Then, the outlier detection method of inertial devices based on extrapolation prediction is added. The predicted value of the inertial device is extrapolated by the fourth-order difference method, and the outliers are recognized and eliminated by the adaptive threshold, which contributes to improving the robustness and accuracy of array POS. STD is selected in this paper to statistic the accuracy of array POS. Compared with the traditional federated Kalman filtering (KFK) methods, the accuracies of position, speed, heading angle and horizontal attitude angle of the left node and right node are all improved when there are outliers in the measurement data. Compared with the fault-tolerant federated combination method based on innovation characteristics, the accuracies of position, speed, heading angle and horizontal attitude angle of the left node and right node are all improved when there are abnormal values in the inertial device data

On-Demand Fully Enclosed Superhydrophobic-Optofluidic Devices Enabled by Microstereolithography.

Superhydrophobic surface-based optofluidics have been introduced to biosensors and unconventional optics with unique advantages, such as low light loss and power consumption. However, most of these platforms were made with planar-like microstructures and nanostructures, which may cause bonding issues and result in significant waveguide loss. Here, we introduce a fully enclosed superhydrophobic-based optofluidics system, enabled by a one-step microstereolithography procedure. Various microstructured cladding designs with a feature size down to 100 μm were studied and a T-type overhang design exhibits the lowest optical loss, regardless of the excitation wavelength. Surprisingly, the optical loss of superhydrophobic-based optofluidics is not solely decided by the solid area fraction at the solid/water/air interface, but also the cross-section shape and the effective cladding layer composition. We show that this fully enclosed optofluidic system can be used for CRISPR-labeled quantum dot quantification, intended for in vitro and in vivo CRISPR therapeutics

Perspective of Molecular Diagnosis in Healthcare: From Barcode to Pattern Recognition

Barcode technology has a broad spectrum of applications including healthcare, food security, and environmental monitoring, due to its ability to encode large amounts of information. With the rapid development of modern molecular research, barcodes are utilized as a reporter with different molecular combinations to label many biomolecular targets, including genomic and metabolic elements, even with multiplex targeting. Along with the advancements in barcoded bioassay, the improvements of various designs of barcode components, encoding and decoding strategies, and their portable adoption are indispensable in satisfying multiple purposes, such as medical confirmation and point-of-care (POC) testing. This perspective briefly discusses the current direction and progress of barcodes development and provides a hypothesis for barcoded bioassay in the near future