Amino acids as stabilizers for lysozyme during the spray-drying process and storage.

Amino acids (AAs) have been used as excipients in protein formulations both in solid and liquid state products due to their stabilizing effect. However, the mechanisms by which they can stabilize a protein have not been fully elucidated yet. The purpose of this study was to investigate the effect of AAs with distinct physicochemical properties on the stability of a model protein (lysozyme, LZM) during the spray-drying process and subsequent storage. Molecular descriptor based multivariate data analysis was used to select distinct AAs from the group of 20 natural AAs. Then, LZM and the five selected AAs (1:1 wt ratio) were spray-dried (SD). The solid form, residual moisture content (RMC), hygroscopicity, morphology, secondary/tertiary structure and enzymatic activity of LZM were evaluated before and after storage under 40 °C/75 % RH for 30 days. Arginine (Arg), leucine (Leu), glycine (Gly), tryptophan (Trp), aspartic acid (Asp) were selected because of their distinct properties by using principal component analysis (PCA). The SD LZM powders containing Arg, Trp, or Asp were amorphous, while SD LZM powders containing Leu or Gly were crystalline. Recrystallization of Arg, Trp, Asp and polymorph transition of Gly were observed after the storage under accelerated conditions. The morphologies of the SD particles vary upon the different AAs formulated with LZM, implying different drying kinetics of the five model systems. A tertiary structural change of LZM was observed in the SD powder containing Arg, while a decrease in the enzymatic activity of LZM was observed in the powders containing Arg or Asp after the storage. This can be attributed to the extremely basic and acidic conditions that Arg and Asp create, respectively. This study suggests that when AAs are used as stabilizers instead of traditional disaccharides, not only do classic vitrification theory and water replacement theory play a role, but the microenvironmental pH conditions created by basic or acidic AAs in the starting solution or during the storage of solid matter are also crucial for the stability of SD protein products.

Time-dependent proteomics and drug response in expanding cancer cells.

Cancer cell line is commonly used for discovery and development of anti-cancer drugs. It is generally considered that drug response remains constant for a certain cell line due to the identity of genetics thus protein patterns. Here, we demonstrated that cancer cells continued dividing even after reaching confluence, in that the proteomics was changed continuously and dramatically with strong relevance to cell division, cell adhesion and cell metabolism, indicating time-dependent intrinsically reprogramming of cells during expansion. Of note, the inhibition effect of most anti-cancer drugs was strikingly attenuated in culture cells along with cell expansion, with the strongest change at the third day when cells were still expanding. Profiling of an FDA-approved drug library revealed that attenuation of response with cell expansion is common for most drugs, an exception was TAK165 that was a selective inhibitor of mitochondrial respiratory chain complex I. Finally, we screened a panel of natural products and identified four pentacyclic triterpenes as selective inhibitors of cancer cells under prolonged growth. Taken together, our findings underscore that caution should be taken in evaluation of anti-cancer drugs using culture cells, and provide agents selectively targeting overgrowth cancer cells.

Bacterium secretes chemical inhibitor that sensitizes competitor to bacteriophage infection.

To overtake competitors, microbes produce and secrete secondary metabolites that kill neighboring cells and sequester nutrients. This natural product-mediated competition likely evolved in complex microbial communities that included viral pathogens. From this ecological context, we hypothesized that microbes secrete metabolites that "weaponize" natural pathogens (i.e., bacteriophages) to lyse their competitors. Indeed, we discovered a bacterial secondary metabolite that sensitizes other bacteria to phage infection. We found that this metabolite provides the producer (a Streptomyces sp.) with a fitness advantage over its competitor (Bacillus subtilis) by promoting phage infection. The phage-promoting metabolite, coelichelin, sensitized B. subtilis to a wide panel of lytic phages, and it did so by preventing the early stages of sporulation through iron sequestration. Beyond coelichelin, other natural products may provide phage-mediated competitive advantages to their producers-either by inhibiting sporulation or through yet-unknown mechanisms.

Development and psychometric assessment of Health Action Process Approach (HAPA) in terms of smoking cessation among Chinese smokers.

The Health Action Process Approach (HAPA) is a two-stage (pre-intentional and post-intentional) behavioral change model that distinguishes between motivation and volition in behavior change process. This study aims to develop HAPA-based assessments for smoking cessation among current smokers. The HAPA-based measures were developed and the draft measures included nine constructs, namely, risk perception in smoking-induced cancer, risk perception in smoking-induced systemic disease, positive outcome expectancy, negative outcome expectancy, self-efficacy in quitting smoking, self-efficacy in maintaining, self-efficacy in re-initiating, quitting planning and coping planning in smoking cessation, with a total of 26 items. A cross-sectional survey was conducted in China in 2022. Principal Component Analysis was used for Exploratory Factor Analysis (EFA). Cronbach's α coefficient was calculated to evaluate the internal consistency. Variables such as severity of smoking addiction were selected to evaluate the correlation between the HAPA scale and these variables. Of the 928 participants, 76.4% (709/928) were male and the median age was 35 years. Five factors were extracted by EFA. The factor loadings of each item were all greater than 0.60, and the cumulative variance contribution rate was 90.15%. The Cronbach's α coefficient of each HAPA-based subscales was 0.929-0.986. The HAPA-based measurements are comprehensive, reliable and valid in the assessment of smokers' smoking cessation cognition, which can be used to guide the design and implementation of intervention and the development of theory.

Long-acting inhaled medicines: Present and future.

Inhaled medicines continue to be an essential part of treatment for respiratory diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis. In addition, inhalation technology, which is an active area of research and innovation to deliver medications via the lung to the bloodstream, offers potential advantages such as rapid onset of action, enhanced bioavailability, and reduced side effects for local treatments. Certain inhaled macromolecules and particles can also end up in different organs via lymphatic transport from the respiratory epithelium. While the majority of research on inhaled medicines is focused on the delivery technology, particle engineering, combination therapies, innovations in inhaler devices, and digital health technologies, researchers are also exploring new pharmaceutical technologies and strategies to prolong the duration of action of inhaled drugs. This is because, in contrast to most inhaled medicines that exert a rapid onset and short duration of action, long-acting inhaled medicines (LAIM) improve not only the patient compliance by reducing the dosing frequency, but also the effectiveness and convenience of inhaled therapies to better manage patients' conditions. This paper reviews the advances in LAIM, the pharmaceutical technologies and strategies for developing LAIM, and emerging new inhaled modalities that possess a long-acting nature and potential in the treatment and prevention of various diseases. The challenges in the development of the future LAIM are also discussed where active research and innovations are taking place.

Split-Type Magnetic Soft Tactile Sensor with 3D Force Decoupling.

Tactile sensory organs for sensing 3D force, such as human skin and fish lateral lines, are indispensable for organisms. With their sensory properties enhanced by layered structures, typical sensory organs can achieve excellent perception as well as protection under frequent mechanical contact. Here, inspired by these layered structures, a split-type magnetic soft tactile sensor with wireless 3D force sensing and a high accuracy (1.33%) fabricated by developing a centripetal magnetization arrangement and theoretical decoupling model is introduced. The 3D force decoupling capability enables it to achieve a perception close to that of human skin in multiple dimensions without complex calibration. Benefiting from the 3D force decoupling capability and split design with a long effective distance (>20 mm), several sensors are assembled in air and water to achieve delicate robotic operation and water flow-based navigation with an offset <1.03%, illustrating the extensive potential of magnetic tactile sensors in flexible electronics, human-machine interactions, and bionic robots.

The spatial dissimilarities and connections of the microbiota in the upper and lower respiratory tract of beef cattle.

Bovine respiratory disease (BRD) causes morbidity and mortality in cattle. The critical roles of the respiratory microbiota in BRD have been widely studied. The nasopharynx was the most popular sampling niche for BRD pathogen studies. The oral cavity and other niches within the respiratory tract, such as nostrils and lung, are less assessed. In this study, oropharyngeal swabs (OS), nasal swabs (NS), nasopharyngeal swabs (NP), and bronchoalveolar lavage (BAL) were collected from calves located in four countries and analyzed for investigation of the dissimilarities and connections of the respiratory microbiota. The results showed that the microbial diversity, structure, and composition in the upper and lower respiratory tract in beef cattle from China, the USA, Canada, and Italy were significantly different. The microbial taxa for each sampling niche were specific and associated with their local physiology and geography. The signature microbiota for OS, NS, NP, and BAL were identified using the LEfSe algorithm. Although the spatial dissimilarities among the respiratory niches existed, the microbial connections were observed in beef cattle regardless of geography. Notably, the nostril and nasopharynx had more similar microbiomes compared to lung communities. The major bacterial immigration patterns in the bovine respiratory tract were estimated and some of them were associated with geography. In addition, the contribution of oral microbiota to the nasal and lung ecosystems was confirmed. Lastly, microbial interactions were characterized to reveal the correlation between the commercial microbiota and BRD-associated pathogens. In conclusion, shared airway microbiota among niches and geography provides the possibility to investigate the common knowledge for bovine respiratory health and diseases. In spite of the dissimilarities of the respiratory microbiota in cattle, the spatial connections among these sampling niches not only allow us to deeply understand the airway ecosystem but also benefit the research and development of probiotics for BRD.

Crystal Growth of Defect-Free Single-Crystalline Fibers in Super-High Supersaturation Solution.

By controlling the supersaturation and choosing high-quality seeds, we successfully suppress the prismatic growth of the tetragonal potassium dihydrogen phosphate (KDP) SCFs, realize the rapid growth along the [001] direction, and obtain SCFs less than 10 µm in width with lengths of centimeters. The experimental results show that there exist critical supersaturation points, 22.40% and 41.41% at 25 °C, for initiating the growth of KDP SCF on its pyramidal and prismatic faces, respectively, which are quite different from those of the bulk crystals. We use the mechanism of 2D nucleation on smooth faces to explain the peculiar phenomena, assuming that there is no 2D or 3D defect on the surfaces of the seed fiber crystal. The assumption is supported by AFM observation of the surface micromorphology of the SCFs. Our solution growth technique developed can be used to grow ultrafine SCFs unable to be achieved by existing melt growth techniques.

Hybrid Multimaterial 3D Printing Using Photocuring-While-Dispensing.

Three-dimensional (3D) printing methods, such as vat photopolymerization (VPP) and direct-ink-writing (DIW) processes, are known for their high-resolution and multimaterial capabilities, respectively. Here a novel hybrid 3D printing technique that combines the strengths of VPP and DIW processes to achieve multimaterial and high-resolution printing of functional structures and devices, is presented. The method involves dispensing liquid-like materials via syringes into a photocurable matrix material and subsequently using a Galvano mirror-controlled laser beam to selectively photocure the dispensed material trace or the matrix material surrounding the trace. The laser beam scanning and syringe dispensing are synchronized with a set delay to control liquid diffusion and in situ fixture. The versatility of the method is demonstrated by fabricating intricate 3D ant and wheel prototypes using various materials available for VPP and DIW technologies. The proposed photocuring-while-dispensing strategy offers advantages over conventional multimaterial 3D printing methods, such as integrating materials regardless of photocurability and viscosity, and fabricating heterogeneous structures with complex geometries and high resolution. With its principle demonstrated, this multimaterial 3D printing process will open up a wide range of potential applications with diverse functionalities and materials.

Ravidomycin Analogs from Streptomyces sp. Exhibit Altered Antimicrobial and Cytotoxic Selectivity.

Six new ravidomycin analogs (1-4, 6, and 7) were isolated from Streptomyces sp. Am59 using UV- and LCMS-guided separation based on Global Natural Products Social (GNPS) molecular networking analysis. Furthermore, we isolated fucomycin V (9), which possesses the same chromophore as ravidomycin but features a d-fucopyranose instead of d-ravidosamine. This is the first report of 9 as a natural product. Four new analogs (10-13) of 9 were also isolated. The structures were elucidated by combined spectroscopic and computational methods. We also found an inconsistency with the published [α]D25 of deacetylravidomycin, which is reported to have a (-) sign. Instead, we observed a (+) specific rotation for the reported absolute configuration of deacetylravidomycin (containing d-ravidosamine). We confirmed the positive sign by reisolating deacetylravidomycin from S. ravidus and by deacetylating ravidomycin. Finally, antibacterial, antifungal, and cytotoxicity activities were determined for the compounds. Compared to deacetylravidomycin, the compounds 4-6, 9, 11, and 12 exhibited greater antibacterial selectivity.

Study of Preparation and Properties of Stereoregular Poly(cyclohexenylene carbonate).

Fixing carbon dioxide as a polymer material is an effective and environmentally beneficial approach for reducing the harm of CO2 greenhouse gas. In this paper, carbon dioxide and cyclohexene oxide were used as co-monomers, and a chiral binuclear cobalt complex with a biphenyl linker was employed as the catalyst to successfully prepare a poly(cyclohexenylene carbonate) with high stereoregularity. The influence of catalyst structure, CO2 pressure, and operating temperature on the copolymerization rate and polymer structure were systematically investigated. Optimal catalyst structure and operating conditions were determined, resulting in an excellent poly(cyclohexenylene carbonate) with a stereoregularity as high as 93.5%. Performance testing revealed that the polyester had a molecular weight of approximately 20 kg/mol, a glass transition temperature of 129.7 °C, an onset decomposition temperature of 290 °C, and a tensile strength of 42.8 MPa. These results demonstrate high thermal stability and mechanical strength, indicating the potential for expanding the applications of aliphatic polycarbonate materials.

Data-driven prediction of complex crystal structures of dense lithium.

Lithium (Li) is a prototypical simple metal at ambient conditions, but exhibits remarkable changes in structural and electronic properties under compression. There has been intense debate about the structure of dense Li, and recent experiments offered fresh evidence for yet undetermined crystalline phases near the enigmatic melting minimum region in the pressure-temperature phase diagram of Li. Here, we report on an extensive exploration of the energy landscape of Li using an advanced crystal structure search method combined with a machine-learning approach, which greatly expands the scale of structure search, leading to the prediction of four complex Li crystal structures containing up to 192 atoms in the unit cell that are energetically competitive with known Li structures. These findings provide a viable solution to the observed yet unidentified crystalline phases of Li, and showcase the predictive power of the global structure search method for discovering complex crystal structures in conjunction with accurate machine learning potentials.

A Personalized Mobile Cessation Intervention to Promote Smokers From the Preparation Stage to the Action Stage: Double-blind Randomized Controlled Trial.

BACKGROUND: Most mobile cessation studies have found that such interventions have a higher quitting rate than interventions providing minimal smoking cessation support. However, why such interventions are effective has been almost unstudied by researchers. OBJECTIVE: This paper describes the principles of the personalized mobile cessation intervention-based WeChat app and used generalized estimated equations to assess why a personalized mobile cessation intervention was more likely to promote smokers from the preparation stage to the action stage than a nonpersonalized intervention. METHODS: This is a 2-arm, double-blind, randomized controlled trial in five cities in China. The intervention group received a personalized mobile cessation intervention. The control group received a nonpersonalized SMS text message smoking cessation intervention. All information was sent by the WeChat app. The outcomes were the change in protection motivation theory construct scores and the change in transtheoretical model stages. RESULTS: A total of 722 participants were randomly assigned to the intervention or control group. Compared with those who received the nonpersonalized SMS text message intervention, smokers who received the personalized intervention presented lower intrinsic rewards, extrinsic rewards, and response costs. Intrinsic rewards were determinants of stage change, thus explaining why the intervention group was more likely to promote smokers from the preparation stage to the action stage (odds ratio 2.65, 95% CI 1.41-4.98). CONCLUSIONS: This study identified the psychological determinants at different stages to facilitate smokers moving forward to the next stage of quitting behavior and provides a framework to explore why a smoking cessation intervention is effective. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2100041942; https://tinyurl.com/2hhx4m7f.

Room Temperature Construction of Vicinal Amino Alcohols via Electroreductive Cross-Coupling of N-Heteroarenes and Carbonyls.

Despite the widespread applications of α-hydroxyalkyl cyclic amines, direct and diverse access to such a class of unique vicinal amino alcohols still remains, to date, a challenge. Here, through a strategy of electroreductive α-hydroxyalkylation of inactive N-heteroarenes with ketones or electron-rich arylaldehydes, we describe a room temperature approach for the direct construction of α-hydroxyalkyl cyclic amines, which features a broad substrate scope, operational simplicity, high chemoselectivity, and no need for pressurized H2 gas and transition metal catalysts. The zinc ion generated from anode oxidation plays a crucial role in the activation of both reactants by decreasing their reduction potentials. The strategy of electroreduction in combination with substrate activation by Lewis acids in this work is anticipated to develop more useful transformations.

[Microplastic Pollution Status and Ecological Risk Evaluation in Weihe River].

This study aimed to explore the occurrence state, morphological characteristics, polymer types, as well as potential ecology of microplastics in the Weihe River in Northwest China. Thus, we identified the abundance distribution, shape, particle size, color, and polymer type of microplastics in the surface water of the Weihe River by conducting field sampling, microscope observation, Fourier infrared spectroscopy, Micro-Raman spectroscopy, etc., during the normal water period (May) of 2021. Subsequently, we analyzed the potential ecological and environmental risks of microplastics using the pollution load index method and species sensitivity distribution method. The results showed that microplastics were detected to exist in all sampling points and the abundance ranged from (2.9±0.8) to (10.3±2.8) n·L-1. The concentration of microplastics in the main stream of the Weihe River was higher than that in the tributaries. Fiber (15.04%-77.03%), small size (<0.5 mm) (27.27%-89.38%), and colored (15.85%-49.53%) were the predominant microplastic types. Polyethylene (32.98%), polypropylene (29.79%), polystyrene (21.21%), and polyethylene (10.61%) terephthalate were the main types of polymers detected. In general, the microplastic pollution in Weihe River was at a medium level and had not affected aquatic organisms, but its high concentration and the characteristic of the plastics to adsorb other pollutants are still of concern.

Wireless Flexible Magnetic Tactile Sensor with Super-Resolution in Large-Areas.

Tactile recognition is among the basic survival skills of human beings, and advances in tactile sensor technology have been adopted in various fields, bringing benefits such as outstanding performance in manipulating objects and general human-robot interactions. However, promoting enhanced perception of the existing tactile sensors is limited by their sensor array arrangement and wire-connected design. Here we present a wireless flexible magnetic tactile sensor (FMTS) consisting of a multidirection magnetized flexible film (perception module) and a contactless Hall sensor (signal receiving module). The flexible magnetic film is composed of NdFeB microparticles and soft silicone elastomer microparticles, and it transfers the unambiguous transduction of external force position and magnitude into magnetic signals. Benefiting from the specific magnetization arrangement and clustering algorithm, only one Hall sensor is needed in FMTS to perceive the magnitude and position of the contact spot simultaneously with super-resolution (2.1 mm average error) on a large area (3600 mm2), and the effective working distance is also greatly extended (∼30 mm), allowing for the full softness and adaptability to diverse conditions. We anticipate that this design will promote the development of soft tactile sensors and their integration into human-robot interaction and humanoid robot perception.

Biological Functions and Cross-Kingdom Host Gene Regulation of Small RNAs in Lactobacillus plantarum-Derived Extracellular Vesicles.

Extracellular vesicle-mediated transfer of microRNAs is a novel mode of cell-to-cell genetic transmission. Extracellular vesicles produced by microbes have been shown to contain significant quantities of physiologically active molecules such as proteins, lipids, and RNA, which could be transported to host cells and play a key role in both inter-kingdom signaling and physiological responses. In this study, we identified sRNAs by sequencing small RNAs (sRNAs) from Lactobacillus plantarum-derived extracellular vesicles (LDEVs) and detected the expression levels of vesicular sRNAs using quantitative reverse transcription-polymerase chain reaction (RT-PCR), which demonstrated the presence of microRNA-sized RNAs (msRNAs) within these vesicles. We chose sRNA71, a highly expressed msRNA, for further investigation, predicted its potential target genes for the human genome, and indicated that it could be translocated into mammalian cells. The biological functions of this sRNA71 were subsequently explored through cellular proteomics, western blot, and luciferase reporter assay. According to the findings, transfection with synthetic sRNA71 mimics substantially reduced Tp53 expression in HEK293T cells and suppressed the gene expression through binding to the 3' UTR of Tp53 mRNA. In conclusion, it is hypothesized that microbial-derived extracellular vesicles serve as carriers of functional molecules such as sRNAs, which play an essential role in regulating microbial-host communication.

Total Syntheses of Calyciphylline A-Type Alkaloids (-)-10-Deoxydaphnipaxianine A, (+)-Daphlongamine E and (+)-Calyciphylline R via Late-Stage Divinyl Carbinol Rearrangements.

Among the famous Daphniphyllum alkaloids family, the calyciphylline A-type subfamily has triggered particular interest from the organic synthesis community in recent years. Here, we report divergent total syntheses of three calyciphylline A-type alkaloids, namely, (-)-10-deoxydaphnipaxianine A, (+)-daphlongamine E, and (+)-calyciphylline R. Our work highlights an efficient, divergent strategy via late-stage divinyl carbinol rearrangements, including an unprecedented oxidative Nazarov electrocyclization using an unfunctionalized tertiary divinyl carbinol and an unusual allylic alcohol rearrangement. A highly efficient "donor-acceptor" platinum catalyst was used for a critical nitrile hydration step. Moreover, the power of selective amide reductions has also been showcased by novel and classic tactics.

Eltrombopag binds SDC4 directly and enhances MAPK signaling and macropinocytosis in cancer cells.

Syndecan-4 (SDC4) is a single-pass transmembrane glycoprotein implicated in a variety of oncogenic signaling pathways. It is also an intrinsically disordered protein and considered "undruggable". In the present study, we confirmed that knocking out SDC4 in pancreatic cancer cells markedly impaired macropinocytosis, colony formation, as well as xenograft tumor initiation and growth. Quantitative proteomic profiling of Sdc4 knockout (KO) cells revealed significant changes in cell metabolic pathways. In a cellular protein-based ligand interaction screening, we identified that Eltrombopag (ETBP), an FDA-approved agonist of the thrombopoietin receptor (TPOR) for immune thrombocytopenia, could directly bind to SDC4 with a Kd value of ~2 µM. We showed that the transmembrane motif was essential for SDC4 binding to ETBP. Unexpectedly, ETBP not only increased SDC4 abundance, but also enhanced SDC4-associated MAPK signaling pathway and macropinocytosis in cancer cells. Our results indicate that ETBP is a potential agonist of SDC4 in a fashion similar to its original target TPOR, and that caution should be taken when using ETBP for chemotherapy-induced thrombocytopenia in cancer patients.

Magneto-Orientation of Magnetic Double Stacks for Patterned Anisotropic Hydrogels with Multiple Responses and Modulable Motions.

Reported here is a multi-response anisotropic poly(N-isopropylacrylamide) hydrogel developed by using a rotating magnetic field to align magnetic double stacks (MDSs) that are fixed by polymerization. The magneto-orientation of MDSs originates from the unique structure with γ-Fe2 O3 nanoparticles sandwiched by two silicate nanosheets. The resultant gels not only exhibit anisotropic optical and mechanical properties but also show anisotropic responses to temperature and light. Gels with complex ordered structures of MDSs are further devised by multi-step magnetic orientation and photolithographic polymerization. These gels show varied birefringence patterns with potentials as information materials, and can deform into specific configurations upon stimulations. Multi-gait motions are further realized in the patterned gel through dynamic deformation under spatiotemporal light and friction regulation by imposed magnetic force. The magneto-orientation assisted fabrication of hydrogels with anisotropic structures and additional functions should bring opportunities for gel materials in biomedical devices, soft actuators/robots, etc.