Enhancing chiral molecule detection: A weak measurement approach utilizing two-dimensional information.

This study addresses the critical need for high purity chiral molecules in biological systems by overcoming the challenges associated with the quantitative detection of chiral molecules and their enantiomeric mixtures. We developed an innovative detection approach that leverages the two-dimensional information gleaned from natural optical rotation (NOR) and Faraday optical rotation (FOR) under magnetic fields in chiral molecules, combined with an ultrahigh-resolution weak measurement sensor. This novel weak measurement system achieves unparalleled accuracy in detecting spin angles, with a precision of 1.86 × 10-5°. Notably, our method introduces no chemical reactions or interference with the substances under test. It offers enhanced discrimination capabilities through the dual-dimensional analysis of both natural and Faraday optical rotation, alongside a simple and compact sensor design. Conclusively, our study introduces a novel, high-precision, and multi-dimensional optical detection paradigm for chiral molecules. By incorporating Faraday rotation in the presence of a magnetic field, we expand the informational dimensionality accessible to the original weak measurement sensor, facilitating the quantitative analysis of chiral molecules and their enantiomers. This breakthrough not only furnishes a novel instrument for the exploration and development of chiral pharmaceuticals but also propels the advancement of weak measurement sensing technology forward.

Detection of biological loads in sewage using the automated robot-driven photoelectrochemical biosensing platform.

Real-time polymerase chain reaction (RT-PCR) remains the most prevalent molecular detection technology for sewage analysis but is plagued with numerous disadvantages, such as time consumption, high manpower requirements, and susceptibility to false negatives. In this study, an automated robot-driven photoelectrochemical (PEC) biosensing platform is constructed, that utilizes the CRISPR/Cas12a system to achieve fast, ultrasensitive, high specificity detection of biological loads in sewage. The Shennong-1 robot integrates several functional modules, involving sewage sampling and pretreatment to streamline the sewage monitoring. A screen-printed electrode is employed with a vertical graphene-based working electrode and enhanced with surface-deposited Au nanoparticles (NPs). CdTe/ZnS quantum dots (QDs) are further fabricated through the double-stranded DNA (dsDNA) anchored on Au NPs. Using the cDNA template of Omicron BA.5 spike gene as a model, the PEC biosensor demonstrates excellent analytical performance, with a lower detection limit of 2.93 × 102 zm and an outstanding selectivity at the level of single-base mutation recognition. Furthermore, the rapid, accurate detection of BA.5 in sewage demonstrates the feasibility of the PEC platform for sewage monitoring. In conclusion, this platform allows early detection and tracking of infectious disease outbreaks, providing timely data support for public health institutions to take appropriate prevention and control measures.

Ballistic analysis of high-performance armor steel by numerical simulation.

In order to establish a connection between the ballistic performance and mechanical properties of armor steel, a ballistic simulation model was developed and subsequently validated for accuracy and reliability. The mechanical properties of the target plate were described using the Johnson-Cook constitutive relation. An analysis was conducted to investigate the impact of the J-C parameters of the target plate on its ballistic performance, revealing a strong linear relationship between them. Subsequently, a mathematical model represented as H = 14.82 - 0.0048A - 0.0023B + 5.95n - 81.3C was derived, and its accuracy was demonstrated to exceed 90%. This mathematical model can effectively predict the ballistic performance of the armor steel, even when its mechanical properties undergo variations during the production process. This prediction capability significantly contributes to reducing research costs and time.

One-pot hydrothermal synthesis of FeNbO4 microspheres for effective sonocatalysis.

FeNbO4 sonocatalysts were successfully synthesized by a simple hydrothermal route at pH values of 3, 5, 7, 9 and 11. The catalysts were characterized by XRD, XPS, TEM, SEM, N2 adsorption and DRS to analyse the effect of pH parameters on the physicochemical properties of the materials during hydrothermal synthesis. The sonocatalytic activity of FeNbO4 microspheres was evaluated by using acid orange 7 (AO7) as the simulated contaminant. The experimental results showed that the best sonocatalytic degradation ratio (97.45%) of organic dyes could be obtained under the conditions of an initial AO7 concentration of 10 mg L-1, an ultrasonic power of 200 W, a catalyst dosage of 1.0 g L-1, and a pH of 3. Moreover, the sonocatalysts demonstrated consistent durability and stability across multiple test cycles. After active species capture experiments and calculation of the energy band, a possible mechanism was proposed based on the special Fenton-like mechanism and the dissociation of H2O2. This research shows that FeNbO4 microspheres can be used as sonocatalysts for the purification of organic wastewater, which has a promising application prospect.

Effects of Resistant Starch Infusion, Solely and Mixed with Xylan or Cellulose, on Gut Microbiota Composition in Ileum-Cannulated Pigs.

Fermentation of dietary fiber (DF) is beneficial for gut health, but its prebiotic effects are often impeded in the distal large intestine because of the fast degradation of fermentable substrates. One way to enhance the prebiotic effect of DF is to deliver fibers to the lower parts of the gut, which can be achieved by mixing different kinds of fiber. Therefore, in the present study, an ileum-cannulated pig model was employed to investigate the fermentation influence in the large intestine by infusing resistant starch solely (RS, fast fermentable fiber) and mixing with other fibers (xylan or cellulose). Twenty-four ileum-cannulated growing pigs were divided into four groups: one control group receiving saline ileal infusions and three experimental groups infused with RS, RS with xylan, or RS with cellulose. Fecal and plasma samples were analyzed for gut microbiota composition, short-chain fatty acids (SCFAs), and blood biochemistry. Results indicated no significant differences between the RS and control group for the microbiome and SCFA concentration (p > 0.05). However, RS combined with fibers, particularly xylan, resulted in enhanced and prolonged fermentation, marked by an increase in Blautia and higher lactate and acetate production (p < 0.05). In contrast, RS with cellulose infusion enriched bacterial diversity in feces (p < 0.05). Blood biochemistry parameters showed no significant differences across groups (p > 0.05), though a trend of increased glucose levels was noted in the treatment groups (p < 0.1). Overall, RS alone had a limited impact on the distal hindgut microbiota due to rapid fermentation in the proximal gut, whereas combining RS with other fibers notably improved gut microecology by extending the fermentation process.

- Invited Review - Current status, challenges and prospects for pig production in Asia.

Asia is not only the primary region for global pig production but also the largest consumer of pork worldwide. Although the pig production in Asia has made great progress in the past, it still is confronted with numerous challenges. These challenges include: inadequate land and feed resources, a substantial number of small-scale pig farms, escalating pressure to ensure environmental conservation, control of devastating infectious diseases, as well as coping with high temperatures and high humidity. To solve these problems, important investments of human and financial capital are required to promote large-scale production systems, exploit alternative feed resources, implement precision feeding, and focus on preventive medicine and vaccines as alternatives to antibiotics, improve pig breeding, and increase manure recycling. Implementation of these techniques and management practices will facilitate development of more environmentally-friendly and economically sustainable pig production systems in Asia, ultimately providing consumers with healthy pork products around the world.

Patterns of antibiotic administration in Chinese neonates: results from a multi-center, point prevalence survey.

OBJECTIVES: In this study, we describe the patterns of antibiotic prescription for neonates based on World Health Organization's (WHO) Essential Medicines List Access, Watch, and Reserve (AWaRe), and the Management of Antibiotic Classification (MAC) Guidelines in China. METHODS: One-day point-prevalence surveys (PPS) on antimicrobial prescriptions were conducted on behalf of hospitalized neonates in China from September 1 and November 30, annually from 2017 to 2019. RESULTS: Data was collected for a total of 2674 neonatal patients from 15 hospitals in 9 provinces across China of which 1520 were newborns who received at least one antibiotic agent. A total of 1943 antibiotic prescriptions were included in the analysis. The most commonly prescribed antibiotic was meropenem (11.8%). The most common reason for prescribing antibiotic to neonates was pneumonia (44.2%). There were 419 (21.6%), 1343 (69.1%) and 6 (0.3%) antibiotic prescriptions in the Access, Watch and Reserve groups, respectively. According to MAC Guidelines in China, there were 1090 (56.1%) antibiotic agents in the Restricted and 414 (21.3%) in the Special group. CONCLUSION: Broad-spectrum antibiotics included in the Watch and Special groups were likely to be overused in Chinese neonates.

Microstructure and hydrogen storage properties of the Mg2-xYxNi0.9Co0.1 (x = 0, 0.2, 0.3, and 0.4) alloys.

Rare earth elements have excellent catalytic effects on improving hydrogen storage properties of the Mg2Ni-based alloys. This study used a small amount of Y to substitute Mg partially in Mg2Ni0.9Co0.1 and characterized and discussed the effects of Y on the solidification and de-/hydrogenation behaviors. The Mg2-xYxNi0.9Co0.1 (x = 0, 0.2, 0.3, and 0.4) hydrogen storage alloys were prepared using a metallurgy method. The phase composition of the alloys was studied using X-ray diffraction (XRD). Additionally, their microstructure and chemical composition were studied using scanning electron microscopy and energy-dispersive X-ray spectroscopy, respectively. The hydrogen absorption and desorption properties of the alloys were studied using pressure-composition isotherms and differential scanning calorimetric (DSC) measurements. The structure of the as-cast Mg2Ni0.9Co0.1 alloy was composed of the peritectic Mg2Ni, eutectic Mg-Mg2Ni, and a small amount of pre-precipitated Mg-Ni-Co ternary phases, and was converted into the Mg2NiH4, Mg2Ni0.9Co0.1H4, and MgH2 phases after hydrogen absorption. Furthermore, the XRD patterns of the alloys showed the MgYNi4 phase and a trace amount of the Y2O3 phase along with the Mg and Mg2Ni phases after the addition of Y. After hydrogen absorption, the phase of the alloys was composed of the Mg2NiH4, MgH2, MgYNi4, YH3, Y2O3, and Mg2NiH0.3 phases. With the increase of Y addition, the area ratios of the peritectic Mg2Ni matrix phase in the Mg2-xYxNi0.9Co0.1 (x = 0, 0.2, 0.3, and 0.4) alloys gradually decreased until they disappeared. However, the eutectic structure gradually increased, and the microstructures of the alloys were obviously refined. The addition of Y improves the activation performance of the alloys. The alloy only needed one cycle of de-/hydrogenation to complete the activation for x = 0.4. The DSC curves showed that the initial dehydrogenation temperatures of Mg2Ni0.9Co0.1 and Mg1.8Y0.2Ni0.9Co0.1 were 200 and 156 °C, respectively. The desorption activation energies of the hydrides of the Mg2Ni0.9Co0.1 and Mg1.8Y0.2Ni0.9Co0.1 alloys calculated using the Kissinger method were 94.7 and 56.5 kJ/mol, respectively. Moreover, the addition of Y reduced the initial desorption temperature of the alloys and improved their kinetic properties.

Genomic insights into antimicrobial potential and optimization of fermentation conditions of pig-derived Bacillus subtilis BS21.

Bacillus spp. have been widely used as probiotic supplements in animal feed as alternatives to antibiotics. In the present study, we screened a Bacillus subtilis strain named BS21 from pig feces. Antimicrobial activities, whole genome mining and UHPLC-MS/MS analysis were used to explore its antimicrobial mechanism. Strain BS21 showed Significant growth inhibition against a variety of animal pathogens, including Escherichia coli, Salmonella enterica Pullorum, Salmonella enterica Typhimurium, Citrobacter rodentium, Shigella flexneri and Staphylococcus aureus. Seven gene clusters involved in antimicrobial biosynthesis of secondary metabolites were encoded by strain BS21 genome, including four non-ribosomal peptides (bacillibactin, fengycin, surfactin and zwittermicin A), one ribosomal peptide (subtilosin A), one dipeptide (bacilysin) and one polyketide (bacillaene). Among them, production of surfactin, fengycin, bacillibactin, bacilysin and bacillaene was detected in the supernatant of B. subtilis strain BS21. To develop the potential application of BS21 in animal production, medium components and fermentation parameters optimization was carried out using response surface methodology (RSM). Production of antimicrobial secondary metabolites of strain BS21 was increased by 43.4%, and the best medium formula after optimization was corn flour 2%, soybean meal 1.7% and NaCl 0.5% with optimum culture parameters of initial pH 7.0, temperature 30°C, rotating speed at 220 rpm for 26 h. Our results suggested that strain BS21 has the potential for large-scale production and application as a potential source of probiotics and alternative to antibiotics for animal production.

Detecting rare thalassemia in children with anemia using third-generation sequencing.

BACKGROUND: In China, conventional genetic testing methods can only detect common thalassemia variants. Accurate detection of rare thalassemia is crucial for clinical diagnosis, especially for children that need long-term blood transfusion. This study aims to explore the application value of third-generation sequencing (TGS) in the diagnosis of rare thalassemia in children with anemia. METHODS: We enrolled 20 children with anemia, excluding from iron deficiency anemia (IDA). TGS was employed to identify both known and novel thalassemia genotypes, while sanger sequencing was used to confirm the novel mutation detected. RESULTS: Among the 20 samples, we identified 5 cases of rare thalassemia. These included ß-4.9 (hg38,Chr11:5226187-5231089) at HBB gene, α-91(HBA2:c.*91delT), αCD30(HBA2:c.91-93delGAG), Chinese Gγ+(Aγδß)0(NG_000007.3: g .48795-127698 del 78904) and delta - 77(T > C)(HBD:c.-127T>C). Notably, the -SEA/α-91α genotype associated with severe non-deletional hemoglobin H disease (HbH disease) has not been previously reported. Patients with genotypes ß654/ß-4.9 and -SEA/α-91α necessitate long-term blood transfusions, and those with the -SEA/αCD30α, Chinese Gγ+(Aγδß)0 and delta thalassemia demonstrate mild anemia. CONCLUSIONS: TGS demonstrates promising potential as a diagnostic tool for suspected cases of rare thalassemia in children, especially those suspected to have transfusion-dependent thalassemia (TDT).

Endogenous chitinase might lead to differences in growth performance and intestinal health of piglets fed different levels of black soldier fly larva meal.

This study aimed to investigate the effects of different levels of black soldier fly (BSF) replacing soybean meal (SBM) in diets on the performance and health condition of piglets. A total of 180 weaned piglets were allocated into 5 treatments: BSF0 (corn-soybean meal basal diet), BSF25 (BSF replacing 25% SBM), BSF50 (BSF replacing 50% SBM), BSF75 (BSF replacing 75% SBM) and BSF100 (BSF replacing 100% SBM). During the whole period, in comparison with BSF0, average daily gain (ADG) and average daily feed intake increased in the BSF25 and BSF50 groups, whereas ADG decreased in the BSF75 and BSF100 groups (P < 0.05). The result of quadratic fitting curve showed that piglets exhibited the highest ADG when BSF replaced around 20% SBM. Compared with BSF0, organic matter and dry matter digestibility improved in the BSF25 group, whereas ether extract digestibility decreased in the BSF100 group (P < 0.05). In comparison with BSF0, piglets from the BSF25 group showed a higher duodenal ratio of villus height to crypt depth, increased jejunal sucrase activity, serum neuropeptide Y and ghrelin levels, elevated ileal immunoglobulin (Ig) A, IgG and IgM contents and a lower leptin level, and piglets from the BSF100 group exhibited an increased relative weight of kidney (P < 0.05). However, no significant differences were observed in the expression level of tight junction proteins and chitin-degrading enzyme. Additionally, compared with BSF0, the abundance of short chain fatty acid producing bacteria such as Ruminococcaceae, Faecalibacterium and Butyricicoccus increased, and potential pathogenic bacteria decreased in piglets from the BSF25 group, whereas piglets from the BSF100 group had a greater abundance of harmful bacteria. In conclusion, BSF replacing 25% SBM in diets could improve digestive parameters, immune function and intestinal microbiota, and thus improved growth performance of piglets. However, BSF replacing 100% SBM showed an adverse effect on piglet performance, and the reason might be related to the limited amount of chitin-degrading enzyme.

CRISPR-Cas13a-powered electrochemical biosensor for the detection of the L452R mutation in clinical samples of SARS-CoV-2 variants.

Since the end of 2019, a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has deprived numerous lives worldwide, called COVID-19. Up to date, omicron is the latest variant of concern, and BA.5 is replacing the BA.2 variant to become the main subtype rampaging worldwide. These subtypes harbor an L452R mutation, which increases their transmissibility among vaccinated people. Current methods for identifying SARS-CoV-2 variants are mainly based on polymerase chain reaction (PCR) followed by gene sequencing, making time-consuming processes and expensive instrumentation indispensable. In this study, we developed a rapid and ultrasensitive electrochemical biosensor to achieve the goals of high sensitivity, the ability of distinguishing the variants, and the direct detection of RNAs from viruses simultaneously. We used electrodes made of MXene-AuNP (gold nanoparticle) composites for improved sensitivity and the CRISPR/Cas13a system for high specificity in detecting the single-base L452R mutation in RNAs and clinical samples. Our biosensor will be an excellent supplement to the RT-qPCR method enabling the early diagnosis and quick distinguishment of SARS-CoV-2 Omicron BA.5 and BA.2 variants and more potential variants that might arise in the future.

Gut microbiota-derived ursodeoxycholic acid alleviates low birth weight-induced colonic inflammation by enhancing M2 macrophage polarization.

BACKGROUND: Low birth weight (LBW) is associated with intestinal inflammation and dysbiosis after birth. However, the underlying mechanism remains largely unknown. OBJECTIVE: In the present study, we aimed to investigate the metabolism, therapeutic potential, and mechanisms of action of bile acids (BAs) in LBW-induced intestinal inflammation in a piglet model. METHODS: The fecal microbiome and BA profile between LBW and normal birth weight (NBW) neonatal piglets were compared. Fecal microbiota transplantation (FMT) was employed to further confirm the linkage between microbial BA metabolism and intestinal inflammation. The therapeutic potential of ursodeoxycholic acid (UDCA), a highly differentially abundant BA between LBW and NBW piglets, in alleviating colonic inflammation was evaluated in both LBW piglets, an LBW-FMT mice model, and a DSS-induced colitis mouse model. The underlying cellular and molecular mechanisms by which UDCA suppresses intestinal inflammation were also investigated in both DSS-treated mice and a macrophage cell line. Microbiomes were analyzed by using 16S ribosomal RNA sequencing. Fecal and intestinal BA profiles were measured by using targeted BA metabolomics. Levels of farnesoid X receptor (FXR) were knocked down in J774A.1 cells with small interfering RNAs. RESULTS: We show a significant difference in both the fecal microbiome and BA profiles between LBW and normal birth weight animals in a piglet model. Transplantation of the microbiota of LBW piglets to antibiotic-treated mice leads to intestinal inflammation. Importantly, oral administration of UDCA, a major BA diminished in the intestinal tract of LBW piglets, markedly alleviates intestinal inflammation in LBW piglets, an LBW-FMT mice model, and a mouse model of colitis by inducing M2 macrophage polarization. Mechanistically, UDCA reduces inflammatory cytokine production by engaging BA receptor FXR while suppressing NF-κB activation in macrophages. CONCLUSIONS: These findings establish a causal relationship between LBW-associated intestinal abnormalities and dysbiosis, suggesting that restoring intestinal health and postnatal maldevelopment of LBW infants may be achieved by targeting intestinal microbiota and BA metabolism. Video Abstract.

Effects of Avenanthramide on the Small Intestinal Damage through Hsp70-NF-κB Signaling in an Ovalbumin-Induced Food Allergy Model.

A food allergy is caused by an abnormal immune reaction and can induce serious intestinal inflammation and tissue damage. Currently, the avoidance of food allergens is still the most effective way to prevent or reduce allergic symptoms, so the development of new strategies to treat allergies is important. Avenanthramide (AVA) is a bioactive polyphenol derived from oats with a wide range of biological activities; however, it is still not clear whether or how AVA alleviates intestinal damage under allergic situations. The aim of this study was to explore the effect of AVA on the small intestinal damage in an ovalbumin (OVA)-induced food allergy model and its mechanism. In experiment 1, 10 mg/kg bw and 20 mg/kg bw doses of AVA both decreased the serum levels of OVA-specific IgE, histamine, and prostaglandin D induced by OVA. The AVA administration relieved inflammation indicated by the lower serum concentrations of pro-inflammatory cytokines including interleukin-1ß, IL-6, and tumor necrosis factor-α. The levels of tight junction proteins including Claudin-1, ZO-1, and Occludin in the jejunum were elevated after AVA administration, accompanied by the improved intestinal morphology. Furthermore, AVA elevated the protein expression of heat shock protein 70 (Hsp70) and inhibited the phosphorylation of nuclear factor kappa-B (NF-κB), thus the apoptozole, which a Hsp70 inhibitor, was applied in experiment 2 to assess the contribution of Hsp70-NF-κB signaling to the effects of AVA. In the experiment 2, the inhibition of Hsp70 signaling treatment abolished the beneficial effects of AVA on the small intestinal damage and other allergic symptoms in mice challenged with OVA. Taken together, our results indicated that AVA exerted an intestinal protection role in the OVA-induced allergy, the mechanism of which was partly mediated by the Hsp70-NF-κB signaling.

Effects of Protein-Chelated Zinc Combined with Mannan-Rich Fraction to Replace High-Dose Zinc Oxide on Growth Performance, Nutrient Digestibility, and Intestinal Health in Weaned Piglets.

A total of 168 weaned piglets (average initial body weight of 7.70 ± 0.75 kg) were used in a 4-week feeding trial to investigate the effects of dietary supplementation with protein-chelated zinc (Zn-Pro) alone or combined with a mannan-rich fraction (MRF) to replace high-dose zinc oxide (ZnO) for weaned piglets. The dietary treatments included a basal diet as control (CON), a ZnO diet (basal diet + 1600 mg Zn/kg from ZnO), a Zn-Pro diet (basal diet + 60 mg Zn/kg from Zn-Pro), and a MRF plus Zn-Pro diet (MRP, basal diet + 800 mg/kg MRF + 60 mg Zn/kg from Zn-Pro). The average daily gain of piglets in the MRP group was higher (p ≤ 0.05) than that in CON and Zn-Pro groups during d 15-28 and d 1-28 of experiment. The apparent total tract digestibility of dry matter, organic matter, and crude protein in the MRP group was higher (p ≤ 0.05) than that in the CON group. The serum insulin-like growth factor-1 level in the MRP group was markedly higher (p ≤ 0.05) than that of piglets in the other three treatment groups. Piglets fed the Zn-Pro and ZnO diets had greater (p ≤ 0.05) acetic acid in cecal digesta than those fed the CON diet, while the MRP diet had higher (p ≤ 0.05) cecal propionate concentration than those that were fed the CON diet on d 28 of experiment. Moreover, the villus height of ileum in the MRP group tended to be greater than the CON group (p = 0.09). Compared with the CON and MRP groups, the relative abundance of Lactobacillaceae (p = 0.08) and Lachnospiraceae (p = 0.09) in the Zn-Pro group showed an increasing trend. The relative abundance of Prevotellaceae in the Zn-Pro group was significantly lower (p ≤ 0.05) than that in the MRP group. In conclusion, the combined addition of MRF and Zn-Pro acted as a suitable alternative to ZnO to beneficially support the growth performance and intestinal health of weaned piglets, as well as contribute to a lower diarrhea rate and environmental pollution from fecal zinc excretion.

Light-induced tumor theranostics based on chemical-exfoliated borophene.

Among 2D materials (Xenes) which are at the forefront of research activities, borophene, is an exciting new entry due to its uniquely varied optical, electronic, and chemical properties in many polymorphic forms with widely varying band gaps including the lightest 2D metallic phase. In this paper, we used a simple selective chemical etching to prepare borophene with a strong near IR light-induced photothermal effect. The photothermal efficiency is similar to plasmonic Au nanoparticles, with the added benefit of borophene being degradable due to electron deficiency of boron. We introduce this selective chemical etching process to obtain ultrathin and large borophene nanosheets (thickness of ~4 nm and lateral size up to ~600 nm) from the precursor of AlB2. We also report first-time observation of a selective Acid etching behavior showing HCl etching of Al to form a residual B lattice, while HF selectively etches B to yield an Al lattice. We demonstrate that through surface modification with polydopamine (PDA), a biocompatible smart delivery nanoplatform of B@PDA can respond to a tumor environment, exhibiting an enhanced cellular uptake efficiency. We demonstrate that borophene can be more suitable for safe photothermal theranostic of thick tumor using deep penetrating near IR light compared to gold nanoparticles which are not degradable, thus posing long-term toxicity concerns. With about 40 kinds of borides, we hope that our work will open door to more discoveries of this top-down selective etching approach for generating borophene structures with rich unexplored thermal, electronic, and optical properties for many other technological applications.

Extracellular production of Ulp1403-621 in leaky E. coli and its application in antimicrobial peptide production.

Small ubiquitin-like modifier (SUMO) tag is widely used to promote soluble expression of exogenous proteins, which can then be cleaved by ubiquitin-like protease 1 (Ulp1) to obtain interested protein. But the application of Ulp1 in large-scale recombinant protein production is limited by complicated purification procedures and high cost. In this study, we describe an efficient and simple method of extracellular production of Ulp1403-621 using a leaky Escherichia coli BL21(DE3), engineered by deleting the peptidoglycan-associated outer membrane lipoprotein (pal) gene. Ulp1403-621 was successfully leaked into extracellular supernatant by the BL21(DE3)-Δpal strain after IPTG induction. The addition of 1% glycine increased the extracellular production of Ulp1403-621 approximately four fold. Moreover, extracellular Ulp1403-621 without purification had high activities for cleaving SUMO fusion proteins, and antimicrobial peptide pBD2 obtained after cleavage can inhibit the growth of Staphylococcus aureus. The specific activity of extracellular Ulp1403-621 containing 1 mM EDTA and 8 mM DTT reached 2.0 × 106 U/L. Another commonly used protease, human rhinovirus 3C protease, was also successfully secreted by leaky E. coli strains. In conclusion, extracellular production of tool enzymes is an attractive way for producing large-scale active recombinant proteins at a lower cost for pharmaceutical, industrial, and biotechnological applications. KEY POINTS: • First report of extracellular production of Ulp1403-621 in leaky Escherichia coli BL21(DE3) strain. • One percent glycine addition into cultivation medium increased the extracellular production of Ulp1403-621 approximately four fold. • The specific activity of extracellular Ulp1403-621 produced in this study reached 2.0 × 106 U/L.

A CRISPR/Cas12a-empowered surface plasmon resonance platform for rapid and specific diagnosis of the Omicron variant of SARS-CoV-2.

The outbreak of the COVID-19 pandemic was partially due to the challenge of identifying asymptomatic and presymptomatic carriers of the virus, and thus highlights a strong motivation for diagnostics with high sensitivity that can be rapidly deployed. On the other hand, several concerning SARS-CoV-2 variants, including Omicron, are required to be identified as soon as the samples are identified as 'positive'. Unfortunately, a traditional PCR test does not allow their specific identification. Herein, for the first time, we have developed MOPCS (Methodologies of Photonic CRISPR Sensing), which combines an optical sensing technology-surface plasmon resonance (SPR) with the 'gene scissors' clustered regularly interspaced short palindromic repeat (CRISPR) technique to achieve both high sensitivity and specificity when it comes to measurement of viral variants. MOPCS is a low-cost, CRISPR/Cas12a-system-empowered SPR gene-detecting platform that can analyze viral RNA, without the need for amplification, within 38 min from sample input to results output, and achieve a limit of detection of 15 fM. MOPCS achieves a highly sensitive analysis of SARS-CoV-2, and mutations appear in variants B.1.617.2 (Delta), B.1.1.529 (Omicron) and BA.1 (a subtype of Omicron). This platform was also used to analyze some recently collected patient samples from a local outbreak in China, identified by the Centers for Disease Control and Prevention. This innovative CRISPR-empowered SPR platform will further contribute to the fast, sensitive and accurate detection of target nucleic acid sequences with single-base mutations.

Identification of a Novel Hb H Disease with Glucose-6-Phosphate Dehydrogenase Deficiency Using Whole Genome Sequencing.

With the development of sequencing technology, more and more rare thalassemia types have been found. In this article, we found a novel Hb H disease combined with glucose-6-phosphate dehydrogenase (G6PD) deficiency through whole genome sequencing (WGS), which was verified by Sanger sequencing and polymerase chain reaction (PCR)-reverse dot-blot hybridization, respectively.