A high throughput array microhabitat platform reveals how light and nitrogen colimit the growth of algal cells.

A mechanistic understanding of algal growth is essential for maintaining a sustainable environment in an era of climate change and population expansion. It is known that algal growth is tightly controlled by complex interactive physical and chemical conditions. Many mathematical models have been proposed to describe the relation of algal growth and environmental parameters, but experimental verification has been difficult due to the lack of tools to measure cell growth under precise physical and chemical conditions. As such, current models depend on the specific testing systems, and the fitted growth kinetic constants vary widely for the same organisms in the existing literature. Here, we present a microfluidic platform where both light intensity and nutrient gradients can be well controlled for algal cell growth studies. In particular, light shading is avoided, a common problem in macroscale assays. Our results revealed that light and nitrogen colimit the growth of algal cells, with each contributing a Monod growth kinetic term in a multiplicative model. We argue that the microfluidic platform can lead towards a general culture system independent algal growth model with systematic screening of many environmental parameters. Our work advances technology for algal cell growth studies and provides essential information for future bioreactor designs and ecological predictions.

Immune function status of postoperative patients with colon cancer for predicting liver metastasis.

BACKGROUND: Colon cancer (CC) has a high incidence rate. Radical resection is the main treatment method for CC; however, liver metastasis (LM) often occurs post-surgery. The liver contains both innate and adaptive immune cells that monitor and remove abnormal cells and pathogens. Before LM, tumor cells secrete cytokines and exosomes to adjust the immune microenvironment of the liver, thus forming an inhibitory immune microenvironment for colonization by circulating tumor cells. This indicates that the immune state of patients with CC plays a crucial role in the occurrence and progression of LM. AIM: To observe and analyze the relationship between immune status and expression of tumor factors in patients with LM of CC, and to provide a scientific intervention method for promoting the patient prognosis. METHODS: A retrospective analysis was performed. The baseline data of 100 patients with CC and 100 patients with CC who suffered from postoperative LM and were admitted to our hospital from May 2021 to May 2023 were included in the non-occurrence and occurrence groups, respectively. The immune status of the patients and the expression of tumor factor-related indicators in the two groups were compared, and the predictive value of the indicators for postoperative LM in patients with CC was analyzed. RESULTS: Compared with the non-occurrence group, the expression of serum carcinoembryonic antigen (CEA), CA19-9, CA242, CA72-4 and CA50 in patients in the occurrence group were significantly higher, while the expression of CD3+, CD4+, CD8+, natural killer (NK) and CD4+/CD25 in patients in the occurrence group were significantly lower (P < 0.05). No significant difference was observed in other baseline data between groups (P > 0.05). Multivariate logistic regression model analysis revealed that the expressions of CEA, CA19-9, CA242, CA72-4, CA50, CD3+, CD4+, CD8+, NK, and CD4+/CD25 were associated with the LM in patients with CC. High expressions of serum CEA, CA19-9, CA242, CA72-4 and CA50, and low expressions of CD3+, CD4+, CD8+, NK, and CD4+/CD25 in patients with CC were risk factors for LM (OR > 1, P < 0.05). The receiver operating characteristic curve showed that the area under curve for CEA, CA19-9, CA242, CA72-4, CA50, CD3+, CD4+, CD8+, NK, and CD4+/CD25 in the prediction of LM in patients with CC were all > 0.80, with a high predictive value. CONCLUSION: The expression of tumor factors and immune state-related indices in patients with CC is closely associated with the occurrence of LM.

Colimitation of light and nitrogen on algal growth revealed by an array microhabitat platform.

Microalgae are key players in the global carbon cycle and emerging producers of biofuels. Algal growth is critically regulated by its complex microenvironment, including nitrogen and phosphorous levels, light intensity, and temperature. Mechanistic understanding of algal growth is important for maintaining a balanced ecosystem at a time of climate change and population expansion, as well as providing essential formulations for optimizing biofuel production. Current mathematical models for algal growth in complex environmental conditions are still in their infancy, due in part to the lack of experimental tools necessary to generate data amenable to theoretical modeling. Here, we present a high throughput microfluidic platform that allows for algal growth with precise control over light intensity and nutrient gradients, while also performing real-time microscopic imaging. We propose a general mathematical model that describes algal growth under multiple physical and chemical environments, which we have validated experimentally. We showed that light and nitrogen colimited the growth of the model alga Chlamydomonas reinhardtii following a multiplicative Monod kinetic model. The microfluidic platform presented here can be easily adapted to studies of other photosynthetic micro-organisms, and the algal growth model will be essential for future bioreactor designs and ecological predictions.

Transcriptional factor OmpR positively regulates prodigiosin biosynthesis in Serratia marcescens FZSF02 by binding with the promoter of the prodigiosin cluster.

Prodigiosin is a promising secondary metabolite mainly produced by Serratia marcescens. The production of prodigiosin by S. marcescens is regulated by different kinds of regulatory systems, including the EnvZ/OmpR system. In this study, we demonstrated that the regulatory factor OmpR positively regulated prodigiosin production in S. marcescens FZSF02 by directly binding to the promoter region of the prodigiosin biosynthesis cluster with a lacZ reporter assay and electrophoretic mobility shift assay (EMSA). The binding sequence with the pig promoter was identified by a DNase I footprinting assay. We further demonstrate that OmpR regulates its own expression by directly binding to the promoter region of envZ/ompR. For the first time, the regulatory mechanism of prodigiosin production by the transcriptional factor OmpR was revealed.

Microscope-based light gradient generation for quantitative growth studies of photosynthetic micro-organisms.

Photosynthetic micro-organisms are equipped with molecular machineries that are designed to transform light into chemical or bioenergy, and help shape and balance the ecosystem of all life forms on earth. Recently, aquatic ecosystems have been disrupted by climate change, which leads to the frequent occurrence of harmful algal blooms (HABs). HABs endanger drinking water resources and harm the fishing and coastal recreation industries. Despite its urgency, mechanistic understanding of how key biophysical and biochemical parameters impact algal growth is largely unexplored. In this article, we developed a microscope-based light gradient generator for studies of photosynthetic micro-organisms under well-defined light intensity gradients. This technology utilized a commercially available microscope, allowed for controlled light exposure and imaging of cells on the same microscope platform, and can be integrated with any micrometer-scale device. Using this technology, we studied the role of light intensity in the growth of photosynthetic micro-organisms. A parallel study was also carried out using a 96-well plate. Our work revealed that the growth rate of the microalgae/cyanobacteria was significantly regulated by the light intensity and followed Monod or van Oorschot kinetic models. The measured half-saturation constants were compared with those obtained in macro-scale devices, and indicated that shading, light spectrum, and temperature may all play important roles in the light sensitivity of photosynthetic micro-organisms. This work highlighted the importance of analytical tools for quantitative understanding of biophysical parameters in the growth of photosynthetic micro-organisms, and knowledge learned will be critical in the design of future technologies for managing algal blooms or optimizing bioenergy production.

Osteogenic peptides in periodontal ligament stem cell-containing three-dimensional bioscaffolds promote bone healing.

Bone tissue engineering shows great potential in bone regeneration; however, the lack of bone growth factors with high biocompatibility and efficiency is a major concern. Oligopeptides have drawn great attention due to their high biological efficacy, low toxicity, and low molecular weight. The oligopeptide SDSSD promotes the osteogenesis of human periodontal ligament stem cells (hPDLSCs) in vitro. The SDSSD-modified three-dimensional (3D) bioscaffolds promote osteogenesis and bone formation in the subcutaneous pockets of BALB/c nude mice and facilitate bone healing in vivo. Mechanistically, SDSSD promoted bone formation by binding to G protein-coupled receptors and regulating the AKT signaling pathway. 3D-printing bioscaffolds with SDSSD may be potential bone tissue engineering materials for treating bone defects.

Identification of Essential Genes Associated With Prodigiosin Production in Serratia marcescens FZSF02.

Prodigiosin is a promising secondary metabolite produced mainly by Serratia strains. To study the global regulatory mechanism of prodigiosin biosynthesis, a mutagenesis library containing 23,000 mutant clones was constructed with the EZ-Tn5 transposon, and 114 clones in the library showed altered prodigiosin production ability. For 37 of the 114 clones, transposon insertion occurred on the prodigiosin biosynthetic cluster genes; transposon inserted genes of the 77 clones belonged to 33 different outside prodigiosin biosynthetic cluster genes. These 33 genes can be divided into transcription-regulating genes, membrane protein-encoding genes, and metabolism enzyme-encoding genes. Most of the genes were newly reported to be involved in prodigiosin production. Transcriptional levels of the pigA gene were significantly downregulated in 22 mutants with different inserted genes, which was in accordance with the phenotype of decreased prodigiosin production. Functional confirmation of the mutant genes involved in the pyrimidine nucleotide biosynthesis pathway was carried out by adding orotate and uridylate (UMP) into the medium. Gene complementation confirmed the regulatory function of the EnvZ/OmpR two-component regulatory system genes envZ and ompR in prodigiosin production.

A Network Pharmacology to Explore the Mechanism of Calculus Bovis in the Treatment of Ischemic Stroke.

BACKGROUND: Calculus Bovis is a valuable Chinese medicine, which is widely used in the clinical treatment of ischemic stroke. The present study is aimed at investigating its target and the mechanism involved in ischemic stroke treatment by network pharmacology. METHODS: Effective compounds of Calculus Bovis were collected using methods of network pharmacology and using the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM) and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Potential compound targets were searched in the TCMSP and SwissTargetPrediction databases. Ischemic stroke-related disease targets were searched in the Drugbank, DisGeNet, OMIM, and TTD databases. These two types of targets were uploaded to the STRING database, and a network of their interaction (PPI) was built with its characteristics calculated, aiming to reveal a number of key targets. Hub genes were selected using a plug-in of the Cytoscape software, and Gene Ontology (GO) biological processes and pathway enrichment analyses of Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted using the clusterProfiler package of R language. RESULTS: Among 12 compounds, deoxycorticosterone, methyl cholate, and biliverdin were potentially effective components. A total of 344 Calculus Bovis compound targets and 590 ischemic stroke targets were found with 92 overlapping targets, including hub genes such as TP53, AKT, PIK2CA, MAPK3, MMP9, and MMP2. Biological functions of Calculus Bovis are associated with protein hydrolyzation, phosphorylation of serine/threonine residues of protein substrates, peptide bond hydrolyzation of peptides and proteins, hydrolyzation of intracellular second messengers, antioxidation and reduction, RNA transcription, and other biological processes. CONCLUSION: Calculus Bovis may play a role in ischemic stroke by activating PI3K-AKT and MAPK signaling pathways, which are involved in regulating inflammatory response, cell apoptosis, and proliferation.

The Highly Uniform Photoresponsivity from Visible to Near IR Light in Sb2Te3 Flakes.

Broadband photosensors have been widely studied in various kinds of materials. Experimental results have revealed strong wavelength-dependent photoresponses in all previous reports. This limits the potential application of broadband photosensors. Therefore, finding a wavelength-insensitive photosensor is imperative in this application. Photocurrent measurements were performed in Sb2Te3 flakes at various wavelengths ranging from visible to near IR light. The measured photocurrent change was insensitive to wavelengths from 300 to 1000 nm. The observed wavelength response deviation was lower than that in all previous reports. Our results show that the corresponding energies of these photocurrent peaks are consistent with the energy difference of the density of state peaks between conduction and valence bands. This suggests that the observed photocurrent originates from these band structure peak transitions under light illumination. Contrary to the most common explanation that observed broadband photocurrent carrier is mainly from the surface state in low-dimensional materials, our experimental result suggests that bulk state band structure is the main source of the observed photocurrent and dominates the broadband photocurrent.

Microfluidic and mathematical modeling of aquatic microbial communities.

Aquatic microbial communities contribute fundamentally to biogeochemical transformations in natural ecosystems, and disruption of these communities can lead to ecological disasters such as harmful algal blooms. Microbial communities are highly dynamic, and their composition and function are tightly controlled by the biophysical (e.g., light, fluid flow, and temperature) and biochemical (e.g., chemical gradients and cell concentration) parameters of the surrounding environment. Due to the large number of environmental factors involved, a systematic understanding of the microbial community-environment interactions is lacking. In this article, we show that microfluidic platforms present a unique opportunity to recreate well-defined environmental factors in a laboratory setting in a high throughput way, enabling quantitative studies of microbial communities that are amenable to theoretical modeling. The focus of this article is on aquatic microbial communities, but the microfluidic and mathematical models discussed here can be readily applied to investigate other microbiomes.

High-responsivity broad-band sensing and photoconduction mechanism in direct-Gap α-In2Se3 nanosheet photodetectors.

Photoconductivities (PCs) with high responsivity in two-dimensional (2D) diindium triselenide (In2Se3) nanostructures with α-phase hexagonal structure were studied. The In2Se3 nanosheet photodetectors fabricated by focused-ion beam technique exhibit broad spectral response with wavelength range from 300 nm to 1000 nm. The In2Se3 nanosheets achieve optimal responsivity of 720 A W-1 in near-infrared region (808 nm), and detectivity of 2.2 × 1012 Jones, which were higher than several 2D material photodetectors. The physical origins that result in high photoresponse in In2Se3 nanosheets such as carrier lifetime and mobility were also characterized by time-resolved PC and field-effect transistor measurements. The fast (hundred microseconds to milliseconds) and slow (seconds and longer) current rise or decay processes were both observed during the photoresponse. The narrowing (or relaxation) of depletion region and oxygen-sensitized photoconduction mechanism were suggested to be the causes of the efficient photoresponse in the In2Se3 nanostructure detectors. All these observations suggest that α-In2Se3 nanosheets could be a promising candidate for photosensitive material applications.

An array microhabitat device with dual gradients revealed synergistic roles of nitrogen and phosphorous in the growth of microalgae.

Harmful algal blooms (HABs) are an emerging environmental problem contaminating water resources and disrupting the balance of the ecosystems. HABs are caused by the sudden growth of photosynthetic algal cells in both fresh and marine water, and have been expanding in extent and appearing more frequently due to the climate change and population growth. Despite the urgency of the problem, the exact environmental conditions that trigger HABs are unknown. This is in part due to the lack of high throughput tools for screening environmental parameters in promoting the growth of photosynthetic microorganisms. In this article, we developed an array microhabitat device with well defined dual nutrient gradients suitable for quantitative studies of multiple environmental parameters in microalgal cell growth. This device enabled an ability to provide 64 different nutrient conditions [nitrogen (N), phosphorous (P), and N : P ratio] at the same time, and the gradient generation took less than 90 min, advancing the current pond and test tube assays in terms of time and cost. Using a photosynthetic algal cell line, Chlamydomonas reinhardtii, preconditioned in co-limited media, we revealed that N and P synergistically promoted cell growth. Interestingly, no discernible response was observed when single P or N gradient was imposed. Our work demonstrated the enabling capability of the microfluidic platform for screening effects of multiple environmental factors in photosynthetic cell growth, and highlighted the importance of the synergistic roles of environmental factors in algal cell growth.

Apigenin Attenuates Acetaminophen-Induced Hepatotoxicity by Activating AMP-Activated Protein Kinase/Carnitine Palmitoyltransferase I Pathway.

Overuse of acetaminophen (APAP) is a major cause of drug-induced liver failure at the clinics. Apigenin (API) is a natural flavonoid derived from Matricaria chamomilla. The aim of the present study was to investigate the amelioration function of API in APAP-induced hepatotoxicity both in vitro and in vivo and investigate its potential mechanisms. Analysis results of the activities of serum alanine and aspartate aminotransferases (ALT and AST), malondialdehyde, myeloperoxidase (MPO), and reactive oxygen species (ROS) demonstrated therapeutic effects of API. MTT assay results revealed that API attenuated APAP and its metabolic product, N-acetyl-p-benzoquinone imine (NAPQI) induced cytotoxicity in a dose-dependent manner in human liver cells, L-02 cells. Subsequently, metabolomic results of cells and serum analyses demonstrated an aberrant level of carnitine palmitoyltransferase I (CPT1A). We established that API stimulated CPT1A activity in mice liver tissues and L-02 cells. Molecular docking analyses revealed potential interaction of API with CPT1A. Further investigation of the role of CPT1A in L0-2 cells revealed that API reversed cytotoxicity via the AMP-activated protein kinase (AMPK)/GSK-3ß signaling pathway and compound C, which is a selective AMPK inhibitor, inhibited activation of CPT1A induced by API. API was bound to the catalytic region of AMPK as indicated by molecular docking results. In addition, compound C suppressed nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2) that is enhanced by API and inhibited the antioxidative function of API. In summary, the study demonstrates that API attenuates APAP-induced hepatotoxicity by activating the AMPK/GSK-3ß signaling pathway, which subsequently promotes CPT1A activity and activates the NRF2 antioxidant pathway.

Heat Source Forecast of Ball Screw Drive System Under Actual Working Conditions Based on On-Line Measurement of Temperature Sensors.

In view of the time-varying complexity of the heat source for the ball screw feed system, this paper proposes an adaptive inverse problem-solving method to estimate the time-varying heat source and temperature field of the feed system under working conditions. The feed system includes multiple heat sources, and the rapid change of the moving heat source increases the difficulty of its identification. This paper attempts to develop a numerical calculation method for identifying the heat source by combining the experiment with the optimization algorithm. Firstly, based on the theory of heat transfer, a new dynamic thermal network model was proposed. The temperature data signal and the position signal of the moving nut captured by the sensors are used as input to optimize the solution of the time-varying heat source. Then, based on the data obtained from the experiment, finite element software parametric programming was used to optimize the estimate of the heat source, and the results of the two heat source prediction methods are compared and verified. The other measured temperature points obtained by the experiment were used to compare and verify the inverse method of this numerical calculation, which illustrates the reliability and advantages of the dynamic thermal network combined with the genetic algorithm for the inverse method. The method based on the on-line monitoring of temperature sensors proposed in this paper has a strong application value for heat source and temperature field estimation of complex mechanical structures.

Bioinformatic analysis of long non-coding RNA-associated competing endogenous RNA network in adrenocortical carcinoma.

BACKGROUND: Adrenocortical carcinoma (ACC) is a malignant tumor with poor prognosis and unclear pathogenesis. This study aimed to explore the role of long non-coding RNAs (lncRNAs) in ACC. METHODS: We obtained the lncRNA expression profiles of 10 ACC samples and 6 normal control samples from the GEO database and identified differentially expressed RNAs using the limma package in R. RESULTS: We obtained a total of 391 differentially expressed lncRNAs (DElncRNAs) and 1,313 differentially expressed mRNAs (DEmRNAs) between ACC samples and normal control samples. Using Cytoscape v3.7.0, we then constructed a lncRNA-miRNA-mRNA (competing endogenous RNA, or ceRNA) network consisting of 87 lncRNAs, 31 miRNAs, and 78 mRNAs. Applying GO and KEGG enrichment analysis for 78 mRNAs in the ceRNA network, we identified 9 GO terms and 21 significantly enriched pathways. A PPI network was constructed using STRING online tools and Cytoscape v3.7.0, identifying 10 key genes. Finally, through Kaplan-Meier survival analysis, we identified five lncRNAs (LINC00887, MEIS1-AS2, MIR29B2CHG, MIR503HG, and SREBF2-AS1) associated with prognosis in patients with ACC. CONCLUSIONS: In summary, we constructed a ceRNA network and propose a new method for lncRNA research in ACC. Our results provide new clues for further exploration of lncRNAs in the pathogenesis of ACC.

Synthesis of highly fluorescent glutathione-capped Zn(x)Hg(1-x)Se quantum dot and its application for sensing copper ion.

A one-step synthesis of glutathione-capped Zn(x)Hg(1-x)Se quantum dots (QDs) has been accomplished by reacting a mixture of Zn(ClO(4))(2) and Hg(ClO(4))(2) with NaHSe and using glutathione as surface-stabilizing agent. The fluorescent color of the alloyed QDs could be tuned by varying the Zn(2+):Hg(2+) molar ratio, reaction pH, intrinsic Zn(2+) and Hg(2+) reactivity toward NaHSe, and the concentration of NaHSe. The size, composition, and inner structures of these QDs are characterized using dynamic light scattering, inductively coupled plasma-mass spectrometry, and X-ray powder diffraction. By varying the molar ratio of Zn(2+) to Hg(2+) in the precursor solution, the Zn(x)Hg(1-x)Se QDs exhibited a tunable PL wavelength in the range of 548-621 nm. These QDs offered advantages of short reaction time (1 h), low reaction temperature (95 degrees C), high quantum yield, water solubility and biocompatibility. Additionally, glutathione-capped Zn(0.96)Hg(0.04)Se QDs (quantum yield=78%) have been applied for sensing Cu(2+).

Synthesis of Cysteine-Capped Zn(x)Cd(1)(-)(x)Se alloyed quantum dots emitting in the blue-green spectral range.

Alloyed ZnxCd1-xSe quantum dots (QDs) have been successfully prepared at low temperatures by reacting a mixture of Cd(ClO4)2 and Zn(ClO4)2 with NaHSe using cysteine as a surface-stabilizing agent. The photoluminescence (PL) spectra of the alloyed QDs are determined on the basis of the Zn2+/Cd2+ molar ratio, reaction pH, intrinsic Zn2+and Cd2+ reactivities toward NaHSe, concentration of NaHSe, and the kind of thiols. A systematic blue shift in emission wavelength of the alloyed QDs was found with the increase in the Zn mole fraction. This result provides clear evidence of the formation of ZnxCd1-xSe QDs by the simultaneous reaction of Zn2+ and Cd2+ with NaHSe, rather than the formation of separate CdSe and ZnSe nanocrystals or core-shell structure CdSe/ZnSe nanocrystals. The size and inner structure of these QDs are also corroborated by using high-resolution transmission electron microscopy and X-ray powder diffraction. To further understand the formation mechanism, the growth kinetics of Zn0.99Cd0.01Se was studied by measuring the PL spectra at different growth intervals. The results demonstrated that, in the initial stage of growth, Zn0.99Cd0.01Se has a structure with a Cd-rich core and a Zn-rich shell. The post-preparative irradiation of these QDs improved their PL properties, resulting in stronger emission.