CALD1 facilitates epithelial-mesenchymal transition progression in gastric cancer cells by modulating the PI3K-Akt pathway.

BACKGROUND: CALD1 has been discovered to be abnormally expressed in a variety of malignant tumors, including gastric cancer (GC), and is associated with tumor progression and immune infiltration; however, the roles and mechanisms of CALD1 in epithelial-mesenchymal transition (EMT) in GC are unknown. AIM: To investigate the role and mechanism of CALD1 in GC progression, invasion, and migration. METHODS: In this study, the relationship between CALD1 and GC, as well as the possible network regulatory mechanisms of CALD1, was investigated by bioinformatics and validated by experiments. CALD1-siRNA was synthesized and used to transfect GC cells. Cell activity was measured using the CCK-8 method, cell migration and invasive ability were measured using wound healing assay and Transwell assay, and the expression levels of relevant genes and proteins in each group of cells were measured using qRT-PCR and Western blot. A GC cell xenograft model was established to verify the results of in vitro experiments. RESULTS: Bioinformatics results showed that CALD1 was highly expressed in GC tissues, and CALD1 was significantly higher in EMT-type GC tissues than in tissues of other types of GC. The prognosis of patients with high expression of CALD1 was worse than that of patients with low expression, and a prognostic model was constructed and evaluated. The experimental results were consistent with the results of the bioinformatics analysis. The expression level of CALD1 in GC cell lines was all higher than that in gastric epithelial cell line GES-1, with the strongest expression found in AGS and MKN45 cells. Cell activity was significantly reduced after CALD1-siRNA transfection of AGS and MKN45 cells. The ability of AGS and MKN45 cells to migrate and invade was reduced after CALD1-siRNA transfection, and the related mRNA and protein expression was altered. According to bioinformatics findings in GC samples, the CALD1 gene was significantly associated with the expression of members of the PI3K-AKT-mTOR signaling pathway as well as the EMT signaling pathway, and was closely related to the PI3K-Akt signaling pathway. Experimental validation revealed that upregulation of CALD1 increased the expression of PI3K, p-AKT, and p-mTOR, members of the PI3K-Akt pathway,while decreasing the expression of PTEN; PI3K-Akt inhibitor treatment decreased the expression of PI3K, p-AKT, and p-mTOR in cells overexpressing CALD1 (still higher than that in the normal group), but increased the expression of PTEN (still lower than that in the normal group). CCK-8 results revealed that the effect of CALD1 on tumor cell activity was decreased by the addition of the inhibitor. Scratch and Transwell experiments showed that the effect of CALD1 on tumor cell migration and invasion was weakened by the addition of the PI3K-Akt inhibitor. The mRNA and protein levels of EMT-related genes in AGS and MKN45 cells were greatly altered by the overexpression of CALD1, whereas the effect of overexpression of CALD1 was significantly weakened by the addition of the PI3K-Akt inhibitor. Animal experiments showed that tumour growth was slow after inhibition of CALD1, and the expression of some PI3K-Akt and EMT pathway proteins was altered. CONCLUSION: Increased expression of CALD1 is a key factor in the progression, invasion, and metastasis of GC, which may be associated with regulating the PI3K-Akt pathway to promote EMT.

Development of a self-reinforced starch-derived film with biocompatibility and mechanical properties.

The scraps produced while processing packaging materials will cause a waste of resources. In this study, starch-based self-reinforced film (SSRF) using thermoplastic starch (TPS, 45 wt%) and polypropylene (PP, 53 wt%) was developed. The effect of extrusion times (1-4 times) on the film structure and performance was explored. The results show as the number of extrusions increases, the color of SSRF deepens from gray-white to brown, and the crystallinity increases. The mechanical properties of the four types of SSRF first increase and then decrease. The 2-SSRF has the best performance, with tensile strength of 13.23 MPa, elongation at break of 61.35%, Young's modulus of 1128.99 MPa, and flexural strength of 33.19 MPa. Proper extrusion improves the compatibility of TPS and PP. However, repeated extrusion will cause PP degradation and TPS carbonization, reducing interfacial interaction. This study developed new starch-based self-reinforced film and provided theoretical guidance for reusing packaging material scraps.

Extraction of naturally occurring peptides versus the tryptic digestion of proteins from fetal versus adult bovine serum for LC-ESI-MS/MS.

The naturally occurring peptides and digested proteins of fetal versus adult bovine serum were compared by LC-ESI-MS/MS after correction against noise from blank injections and random MS/MS spectra as statistical controls. Serum peptides were extracted by differential precipitation with mixtures of acetonitrile and water. Serum proteins were separated by partition chromatography over quaternary amine resin followed by tryptic digestion. The rigorous X!TANDEM goodness of fit algorithm that has a low error rate as demonstrated by low FDR q-values (q ≤ 0.01) showed qualitative and quantitative agreement with the SEQUEST cross correlation algorithm on 12,052 protein gene symbols. Tryptic digestion provided a quantitative identification of the serum proteins where observation frequency reflected known high abundance. In contrast, the naturally occurring peptides reflected the cleavage of common serum proteins such as C4A, C3, FGB, HPX, A2M but also proteins in lower concentration such as F13A1, IK, collagens and protocadherins. Proteins associated with cellular growth and development such as actins (ACT), ribosomal proteins like Ribosomal protein S6 (RPS6), synthetic enzymes and extracellular matrix factors were enriched in fetal calf serum. In contrast to the large literature from cord blood, IgG light chains were absent from fetal serum as observed by LC-ESI-MS/MS and confirmed by ELISA.

Starch-based biodegradable composites: Effects of in-situ re-extrusion on structure and performance.

In this study, starch-based biodegradable composites (SDC) were prepared by extruding using thermoplastic starch (TPS, 65%wt), polylactic acid (PLA, 30%wt) and poly (butylene adipate co-terephthalate) (PBAT, 5%wt). Structure and properties of the SDC were compared by performing 1-, 2-, 3-times extrusion. The results show that in-situ re-extrusion refines the TPS in composites and reduces the size of the phase. As the number of extrusions increases, the ester bond of composites at 868 cm-1 disappears, the crystallinity increases, and the thermal stability decreases. Among the three types of composites, the mechanical properties and hydrophobic properties of the material obtained by the 2-times are the most outstanding. Compared with SDC, the elongation at break and Young's modulus of SDC-2 are significantly increased, with an increase of 8.01 % and 1.28 % in the machine direction and an increase of 11.02 % and 1.79 % in the transverse direction respectively. Additionally, water contact angle range of SDC-2 from 98.7° to 101.7°. Therefore, SDC prepared by 2-times in-situ re-extrusion has the best film properties and is an ideal packaging material. This study presents a novel method for fabricating starch-degradable composite films by in-situ re-extrusion, providing new insights into the development of starch packaging materials.

Metabolic Engineering of Escherichia coli for High-Titer Biosynthesis of 3'-Sialyllactose.

3'-Sialyllactose (3'-SL) is among the foremost and simplest sialylated breast milk oligosaccharides. In this study, an engineered Escherichia coli for high-titer 3'-SL biosynthesis was developed by introducing a multilevel metabolic engineering strategy, including (1) the introduction of precursor CMP-Neu5Ac synthesis pathway and high-performance α2,3-sialyltransferase (α2,3-SiaT) genes into strain BZ to achieve de novo synthesis of 3'-SL; (2) optimizing the expression of glmS-glmM-glmU involved in the UDP-GlcNAc and CMP-Neu5Ac synthesis pathways, and constructing a glutamine cycle system, balancing the precursor pools; (3) analysis of critical intermediates and inactivation of competitive pathway genes to redirect carbon flux to 3'-SL biosynthesis; and (4) enhanced catalytic performance of rate-limiting enzyme α2,3-SiaT by RBS screening, protein tag cloning. The final strain BZAPKA14 yielded 9.04 g/L 3'-SL in a shake flask. In a 3 L bioreactor, fed-batch fermentation generated 44.2 g/L 3'-SL, with an overall yield and lactose conversion of 0.53 g/(L h) and 0.55 mol 3'-SL/mol, respectively.

Minced Beef Meat Paste Characteristics: Gel Properties, Water Distribution, and Microstructures Regulated by Medium Molecular Mass of γ-Poly-Glutamic Acid.

The influences of various m-γ-PGA (0.08-0.20%, w/w) concentrations on the properties of minced beef meat paste in terms of rheological properties, texture, moisture distribution, and microstructures were evaluated. The results indicated that m-γ-PGA enhanced the water-holding capacity, gel strength, texture, and whiteness of the minced beef meat paste. Based on the microstructural results, m-γ-PGA helped form a more organized and compact gel, thereby limiting the migration of water through the gel matrix. In contrast to the control group, the water-holding property, gel strength, and whiteness of minced meat paste gels with m-γ-PGA content of 0.12% increased from 75.89%, 584.51 g·cm, and 61.83 to 79.91%, 780.87 g·cm, and 62.54, respectively (p < 0.05), exhibiting the highest water-holding property and gel strength. Thus, m-γ-PGA exhibits great potential for minced meat paste products as a healthy gel water retainer and enhancer in low-fat meat products.

Structure and functional characteristics of starch from different hulled oats cultivated in China.

This work aimed to evaluate the structure and functional characteristics of starch from ten hulled oat cultivars grown in different locations in China. The protein, phosphorus, amylose, and starch contents were 0.2-0.4 %, 475.7-691.8 ppm, 16.2-23.0 %, and 93.6-96.7 %, respectively. All the starches showed irregular polygonal shapes and A-type crystallization with molecular weights ranging from 7.2 × 107 to 4.5 × 108 g/mol. The amounts of amylopectin A (DP 6-12), B1 (DP 13-24), B2 (DP 25-36), and B3 (DP > 36) chains were in the ranges of 10.3-16.0 %, 54.5-64.8 %, 16.5-21.1 %, and 4.9-13.1 %, respectively. The starches differed significantly in gelatinization temperatures, pasting viscosity, solubility, swelling power, rheological properties, and digestion parameters. The results revealed that the larger particle size could increase the peak viscosity of the starch paste. The presence of phosphorus increased the gelatinization temperature and enhanced the resistant starch content. The starch granules with higher crystallinity contained a higher proportion of phosphate, which increased final viscosity and setback viscosity but decreased rapidly digestible starch. Overall, oat starch with a high phosphorus content could be used to prepare low-glycemic-index food for diabetes patients.

Expectations for employing Escherichia coli Nissle 1917 in food science and nutrition.

As a promising probiotic strain, Escherichia coli Nissle 1917 (EcN) has been demonstrated to confer beneficial effects on intestinal health, immune function, and pathogen prevention. Additionally, EcN has also been widely studied due to its clear genomic information, tractable gene regulation, and simple growth conditions. This review summarizes the various applications potential of EcN in food science and nutrition, including inflammation prevention, tumor-targeting therapy, antibacterial agents for food, and nutrient production with a focus on specific case studies. Moreover, we highlight the major challenges of employing EcN in food science and nutrition, including regulatory approval, stability during food processing, and consumer acceptance. Finally, we conclude with a discussion on perspectives related to employing EcN in food science and nutrition.

Semi-rationally designed site-saturation mutation of Helicobacter pylori α-1,2-fucosyltransferase for improved catalytic activity and thermostability.

Helicobacter pylori HpfutC, a glycosyltransferase (GT) 11 family glycoprotein, has great potential for industrial 2'-fucosyllactose (2'-FL) production. However, its limited catalytic activity, low expression, and poor thermostability hinder practical applications. Herein, a semi-rationally designed site-saturation mutation was applied to engineer the catalytic activity and thermostability of HpfutC. The 6 single point mutants (K102T, R105C, D115S, Y251F, A255G and K282E) and 6 combined mutants (V1, V2, V3, V4, V5, and V6) with enhanced enzyme activity were obtained by mutant library screening and ordered recombination mutation. The optimal mutant V6, with an optimum temperature of 40 °C, was not a metal-dependent enzyme, yet the reaction was facilitated by Mn2+. Compared to wild-type HpfutC, mutant V6 exhibited a 2.3-fold increase in specific activity and a 2.18-fold increase in half-life at 40 °C, respectively. Kinetic parameters indicated that the Km values of mutant V6 were 34.5 % (lactose) and 25.0 % (GDP-L-fucose) lower than those of the wild enzyme, whereas the kcat/Km values were 1.20 and 1.25-fold higher than those of the wild enzyme. Further, 3D-structure analysis revealed that the highly rigid structure, formation of new hydrogen bonds, increased hydrophobic residues and redistribution of electrostatic charges on the surface may be responsible for the elevated enzyme activity and thermostability. The strategy adopted in this study is of great significance to the solution of the technical bottleneck of HpfutC and the industrial application of 2'-FL.

Zmhdz9, an HD-Zip transcription factor, promotes drought stress resistance in maize by modulating ABA and lignin accumulation.

Maize is the largest crop in the world in terms of both planting area and total yield, and it plays a crucial role in ensuring global food and feed security. However, in recent years, with climate deterioration, environmental changes, and the scarcity of freshwater resources, drought has become a serious limiting factor for maize yield and quality. Drought stress-induced signals undergo a series of transmission processes to regulate the expression of specific genes, thereby affecting the drought tolerance of plants at the tissue, cellular, physiological and biochemical levels. Therefore, in this study we investigated the HD-Zip transcription factor gene Zmhdz9, and yeast activation experiments demonstrated that Zmhdz9 exhibited transcriptional activation activity. Under drought stress, high abscisic acid (ABA) and lignin levels significantly improved drought resistance in maize. Yeast two-hybrid, bimolecular fluorescence complementation (BIFC) and pull-down experiments showed that Zmhdz9 interacted with ZmWRKY120 and ZmTCP9, respectively. Overexpression of Zmhdz9 and gene editing of ZmWRKY120 or ZmTCP9 improved maize drought resistance, indicating their importance in the drought stress response. Furthermore, Zmhdz9 promoted the direct transcription of ZmWRKY120 in the W-box, activating elements of the ZmNCED1 promoter, which encodes a key enzyme in ABA biosynthesis. Additionally, Zmhdz9 promoted direct transcription of ZmTCP9 in the GGTCA motif, activating elements of the ZmKNOX8 promoter, which encodes a key enzyme in lignin synthesis. This study showed that the regulation of ABA and lignin by Zmhdz9 is essential for drought stress resistance in maize.

Characterizations and film-forming properties of different fractionated high-amylose maize starches subjected to hydroxypropylation.

Dual-modifications of jet milling and hydroxypropylation were used to improve the functional properties of maize starch (HM, containing 67 % amylose). The fractions obtained in three sizes (HM-S, HM-M, HM-L) were further treated with 10 % and 30 % propylene oxide (PO10 and PO30). The infrared peak of starch at 2794 cm-1 indicated the successful introduction of hydroxypropyl groups. The molar degree of substitution (MS) increased with the degree of jet milling. The MS of HM-L-PO10 is 0.4, that of HM-M-PO10 is 0.7, and that of HM-S-PO10 is 0.9. The crystallinity of dual-modified HM increased, but the crystal type remained unchanged, still being B-type. Dual-modification significantly improved the performance of starch, and the higher the degree of modification, the better the optimization effect. The lowest enthalpy changes of gelatinization (ΔH = 3.49 J/g), the best freeze-thaw stability, the highest elongation at break (110.42 %) and transmittance (81.22 %) were shown in HM-S-PO30. The present study confirms that HM-S-PO30 films have the best physicochemical and mechanical properties, which provide new insights into optimizing starch-based packaging materials.

Use of airway pressure-based indices to detect high and low inspiratory effort during pressure support ventilation: a diagnostic accuracy study.

BACKGROUND: Assessment of the patient's respiratory effort is essential during assisted ventilation. We aimed to evaluate the accuracy of airway pressure (Paw)-based indices to detect potential injurious inspiratory effort during pressure support (PS) ventilation. METHODS: In this prospective diagnostic accuracy study conducted in four ICUs in two academic hospitals, 28 adult acute respiratory failure patients undergoing PS ventilation were enrolled. A downward PS titration was conducted from 20 cmH2O to 2 cmH2O at a 2 cmH2O interval. By performing an end-expiratory airway occlusion maneuver, the negative Paw generated during the first 100 ms (P0.1) and the maximal negative swing of Paw (∆Pocc) were measured. After an end-inspiratory airway occlusion, Paw reached a plateau, and the magnitude of change in plateau from peak Paw was measured as pressure muscle index (PMI). Esophageal pressure was monitored and inspiratory muscle pressure (Pmus) and Pmus-time product per minute (PTPmus/min) were used as the reference standard for the patient's effort. High and low effort was defined as Pmus > 10 and < 5 cmH2O, or PTPmus/min > 200 and < 50 cmH2O s min-1, respectively. RESULTS: A total of 246 levels of PS were tested. The low inspiratory effort was diagnosed in 145 (59.0%) and 136 (55.3%) PS levels using respective Pmus and PTPmus/min criterion. The receiver operating characteristic area of the three Paw-based indices by the respective two criteria ranged from 0.87 to 0.95, and balanced sensitivity (0.83-0.96), specificity (0.74-0.88), and positive (0.80-0.91) and negative predictive values (0.78-0.94) were obtained. The high effort was diagnosed in 34 (13.8%) and 17 (6.9%) support levels using Pmus and PTPmus/min criterion, respectively. High receiver operating characteristic areas of the three Paw-based indices by the two criteria were found (0.93-0.95). A high sensitivity (0.80-1.00) and negative predictive value (0.97-1.00) were found with a low positive predictive value (0.23-0.64). CONCLUSIONS: By performing simple airway occlusion maneuvers, the Paw-based indices could be reliably used to detect low inspiratory efforts. Non-invasive and easily accessible characteristics support their potential bedside use for avoiding over-assistance. More evaluation of their performance is required in cohorts with high effort.

Human milk oligosaccharide lacto-N-tetraose: Physiological functions and synthesis methods.

Human milk oligosaccharides (HMOs) have attracted considerable attention due to their unique role in boosting infant health. Among the HMOs, lacto-N-tetraose (LNT) is a significant constituent associated with various health benefits, such as prebiotic effects, antiadhesive antimicrobials, antiviral protection, and immune modulators. LNT has received a "Generally Recognized as Safe" status by the American Food and Drug Administration and was approved as a food ingredient for infant formula. However, the limited availability of LNT poses a major challenge for its application in food and medicine. In this review, we first explored the physiological functions of LNT. Next, we describe several synthesis methods for production of LNT, including chemical, enzymatic, and cell factory approaches, and summarize the pivotal research results. Finally, challenges and opportunities for the large-scale synthesis of LNT were discussed.

Development of phytoglycogen-derived core-shell-corona nanoparticles complexed with conjugated linoleic acid.

Phytoglycogen-derived self-assembled nanoparticles (SMPG/CLA) and enzymatic-assembled nanoparticles (EMPG/CLA) were fabricated for delivery of conjugated linoleic acid (CLA). After measuring the loading rate and yield, the optimal ratio for both assembled host-guest complexes was 1 : 10, and the maximum loading rate and yield for EMPG/CLA were 1.6% and 88.1%, respectively, higher than those of SMPG/CLA. Structural characterization studies showed that the assembled inclusion complexes were successfully constructed, and had a specific spatial architecture with inner-core amorphous and external-shell crystalline parts. A higher protective effect against oxidation of EMPG/CLA was observed than that of SMPG/CLA, supporting efficient complexation for a higher order crystalline structure. After 1 h of gastrointestinal digestion under the simulated conditions, 58.7% of CLA was released from EMPG/CLA, which was lower than that released from SMPG/CLA (73.8%). These results indicated that in situ enzymatic-assembled phytoglycogen-derived nanoparticles might be a promising carrier platform for protection and targeted delivery of hydrophobic bioactive ingredients.

Advanced dendritic glucan-derived biomaterials: From molecular structure to versatile applications.

There is considerable interest in the development of advanced biomaterials with improved or novel functionality for diversified applications. Dendritic glucans, such as phytoglycogen and glycogen, are abundant biomaterials with highly branched three-dimensional globular architectures, which endow them with unique structural and functional attributes, including small size, large specific surface area, high water solubility, low viscosity, high water retention, and the availability of numerous modifiable surface groups. Dendritic glucans can be synthesized by in vivo biocatalysis reactions using glucosyl-1-phosphate as a substrate, which can be obtained from plant, animal, or microbial sources. They can also be synthesized by in vitro methods using sucrose or starch as a substrate, which may be more suitable for large-scale industrial production. The large numbers of hydroxyl groups on the surfaces of dendritic glucan provide a platform for diverse derivatizations, including nonreducing end, hydroxyl functionalization, molecular degradation, and conjugation modifications. Due to their unique physicochemical and functional attributes, dendritic glucans have been widely applied in the food, pharmaceutical, biomedical, cosmetic, and chemical industries. For instance, they have been used as delivery systems, adsorbents, tissue engineering scaffolds, biosensors, and bioelectronic components. This article reviews progress in the design, synthesis, and application of dendritic glucans over the past several decades.

Molecular structure and characteristics of phytoglycogen, glycogen and amylopectin subjected to mild acid hydrolysis.

The structure and properties of phytoglycogen and glycogen subjected to acid hydrolysis was investigated using amylopectin as a reference. The degradation took place in two stages and the degree of hydrolysis was in the following order: amylopectin > phytoglycogen > glycogen. Upon acid hydrolysis, the molar mass distribution of phytoglycogen or glycogen gradually shifted to the smaller and broadening distribution region, whereas the distribution of amyopectin changed from bimodal to monomodal shape. The kinetic rate constant for depolymerization of phytoglycogen, amylopectin, and glycogen were 3.45 × 10-5/s, 6.13 × 10-5/s, and 0.96 × 10-5/s, respectively. The acid-treated sample had the smaller particle radius, lower percentage of α-1,6 linkage as well as higher rapidly digestible starch fractions. The depolymerization models were built to interpret the structural differences of glucose polymer during acid treatment, which would provide guideline to improve the structure understanding and precise application of branched glucan with desired properties.

Fabrication of phytoglycogen-derived core-shell nanoparticles: Structure and characterizations.

The objective of this work was to investigate the fabrication of core-shell nanoparticles using phosphorylase-catalyzed chain extension of phytoglycogen, and to analyze the changes of structure and characterizations in detail. During the glucosylation reaction, the inorganic phosphate increased substantially up to 2.3 mg/mL in the initial 12 h, and then increased incrementally to 2.5 mg/mL at 24 h. The similar to trends was observed for increasing Mw and Rz over time, due to glucosyl transfers on the surface chain to form a corona around the phytoglycogen core with a larger size. Phosphorylase modification increases the percentages of longer chain fractions and the average chain length increased from degree of polymerization (DP) 11.6 to DP 48.2. The modified phytoglycogen exhibited the characteristic of B-type crystalline structure, indicating that the specific core-shell nanoparticle with inner amorphous nature and outer crystalline layer. The above results revealed that the potentiality of enzymatic chain elongation of phytoglycogen to design novel core-shell nanoparticle with tailor-made structure and functionality.

Biosynthesis of maltodextrin-derived glucan dendrimer using microbial branching enzyme.

Glucan dendrimers were developed with microbial branching enzyme (BE) treated maltodextrin. The molecular weight (Mw) of recombinant BE was 79.0 kDa, and its optimum activity was observed at pH 7.0 and 70 °C. BE converted different maltodextrins with dextrose equivalent value of 6 (MD6), 12 (MD12), or 19 (MD19) into the given glucan dendrimers, along with the marked increment of the molecular density (approximately 30-60 folds) and α-1,6 linkage percentage (up to 7.3-9.7%). Among three glucan dendrimers, the enzyme-treated MD12 showed a more homogeneous Mw distribution with the maximum Mw of 5.5 × 106 g/mol, indicating that higher substrate catalytic specificity of BE for MD12 substrate. During transglycosylation with MD12 for 24 h, the shorter chains (degree of polymerization, DP < 13) increased from 73.9% to 83.0%, accompanying by a reduction of medium chains (DP13-24) and long chains (DP > 24). Moreover, the slowly digestible and resistant nutritional fractions were increased by 6.2% and 12.5%, respectively. The results suggested that the potentiality of BE structuring glucan dendrimer with tailor-made structure and functionality for industrial application.

Mitochondria and cytochrome components released into the plasma of severe COVID-19 and ICU acute respiratory distress syndrome patients.

INTRODUCTION: Proteomic analysis of human plasma by LC-ESI-MS/MS has discovered a limited number of new cellular protein biomarkers that may be confirmed by independent biochemical methods. Analysis of COVID-19 plasma has indicated the re-purposing of known biomarkers that might be used as prognostic markers of COVID-19 infection. However, multiple molecular approaches have previously indicated that the SARS-COV2 infection cycle is linked to the biology of mitochondria and that the response to infections may involve the action of heme containing oxidative enzymes. METHODS: Human plasma from COVID-19 and ICU-ARDS was analyzed by classical analytical biochemistry techniques and classical frequency-based statistical approaches to look for prognostic markers of severe COVID-19 lung damage. Plasma proteins from COVID-19 and ICU-ARDS were identified and enumerated versus the controls of normal human plasma (NHP) by LC-ESI-MS/MS. The observation frequency of proteins detected in COVID-19 and ICU-ARDS patients were compared to normal human plasma, alongside random and noise MS/MS spectra controls, using the Chi Square (χ2) distribution. RESULTS: PCR showed the presence of MT-ND1 DNA in the plasma of COVID-19, ICU-ARDS, as well as normal human plasma. Mitochondrial proteins such as MRPL, L2HGDH, ATP, CYB, CYTB, CYP, NDUF and others, were increased in COVID-19 and ICU-ARDS plasma. The apparent activity of the cytochrome components were tested alongside NHP by dot blotting on PVDF against a purified cytochrome c standard preparation for H2O2 dependent reaction with luminol as measured by enhanced chemiluminescence (ECL) that showed increased activity in COVID-19 and ICU-ARDS patients. DISCUSSION: The results from PCR, LC-ESI-MS/MS of tryptic peptides, and cytochrome ECL assays confirmed that mitochondrial components were present in the plasma, in agreement with the established central role of the mitochondria in SARS-COV-2 biology. The cytochrome activity assay showed that there was the equivalent of at least nanogram amounts of cytochrome(s) in the plasma sample that should be clearly detectable by LC-ESI-MS/MS. The release of the luminol oxidase activity from cells into plasma forms the basis of a simple and rapid test for the severity of cell damage and lung injury in COVID-19 infection and ICU-ARDS.

Biomimetic synthesis of maltodextrin-derived dendritic nanoparticle and its structural characterizations.

The maltodextrin-derived dendritic nanoparticle was fabricated using microbial branching enzyme and its structural characterizations were investigated. During biomimetic synthesis, molecular weight distribution of maltodextrin substrate with 6.8 × 104 g/mol shifted to the narrower and uniform distribution region with the larger molecular weight up to 6.3 × 106 g/mol (MD12). The enzyme-catalyzed product had the larger size, higher molecular density as well as higher percentage of α-1,6 linkage, accompanying by more chain accumulations of DP 6-12 and disappearance of DP > 24, suggesting the biosynthesized glucan dendrimer had a compact tighter branched structure. The interaction of molecular rotor CCVJ and local structure of dendrimer was monitored, displaying there was a higher intensity related with the numerous nano-pockets at the branch points of MD12. The maltodextrin-derived dendrimers had the single spherical particulate shape with the size range of 10-90 nm. The mathematical models were also established to reveal the chain structuring during enzymatic reaction. The above results showed that the biomimetic strategy for novel dendritic nanoparticle with controllable structure arising from branching enzyme treated maltodextrin, which would help to enlarge the panel of available dendrimer.