Characterization of the key flavor compounds in cream cheese by GC-MS, GC-IMS, sensory analysis and multivariable statistics.

The aroma types of cream cheese affect its commercial value and consumer acceptability. However, the types of volatile substances and sensory characteristics of cream cheese at different fermentation stages are still unclear. Therefore, in this study, headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) and headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS) were used to analyze the volatile substances in cream cheese fermentation. Orthogonal partial least squares discriminant analysis (OPLS-DA), odor activity value (OAV), relative odor activity value (ROAV) and variable projection importance (VIP) were used to identify the characteristic flavor substances in cream cheese fermentation. Finally, the relationship between key flavor substances and sensory characteristics was determined by partial least squares (PLS) analysis. A total of 34 and 36 volatile organic compounds were identified by HS-SPME-GC-MS and HS-GC-MS, respectively, and 14 characteristic flavor substances were found, based on VIP, ROAV and OAV models. Combined with sensory analysis and flavor substance changes, it was found that the cream cheese fermented for 15 d had the best flavor and taste. This study reveals the characteristics and contribution of volatile substances in cream cheese at different fermentation stages, which provides new insights into improving flavor and quality control.

Fatty acid, triglyceride, and kinetic properties of milk fat fractions made by the combination of dry fractionation and short-path molecular distillation.

In this study, we aimed to detect the physicochemical properties of distilled products (residue and distillate) obtained from anhydrous milk fat (AMF) and its dry fractionation products (liquid and solid fractions at 25°C [25 L and 25 S]). The results showed that the saturated fatty acids and low- and medium molecular-weight triglycerides were easily accumulated in the distillate, and the percentage of unsaturated fatty acid and high molecular-weight triglycerides in the residue were higher, and these components in 25 S and 25 L were influenced more significantly than those in the AMF. In addition, the distillate had larger melting ranges in comparison with the distilled substrate, while the melting ranges of residue was smaller. The triglycerides were presented as the mixture crystal forms (α, ß', and ß crystal) in 25 S, AMF, and their distilling products, and it was transformed gradually to a single form as the increasing of distilling temperature. Moreover, the accumulated pattern of triglycerides was double chain length in 25 S, AMF, and their distilling products. These results provide a new approach to obtain the milk fat fractions with different properties, and the findings of this study enrich the theoretical basis of milk fat separation in practical production.

Malolactic Fermentation: New Approaches to Old Problems.

Malolactic fermentation (MLF) is the decarboxylation of L-malic acid to L-lactic acid by lactic acid bacteria (LAB). For the majority of wine production, secondary fermentation is crucial. MLF significantly impacts the quality of most red and some white wine. The outcomes of the spontaneously initiated and finished MLF are frequently unpredictable and can even cause the wine to deteriorate. As a result, individuals typically favour inoculating superior starter cultures when performing MLF. The MLF method for wine has, however, faced new difficulties because of the altered wine fermentation substrate environment brought on by global climate change, the growing demands of winemakers for production efficiency, and the rising demand for high-quality wine. To serve as a reference for the study of wine production and MLF in the current situation, this review primarily updates and summarises the research findings on increasing the effectiveness and dependability of MLF in recent years.

Mechanism and Molecular Targets of Ejiao Siwu Decoction for Treating Primary Immune Thrombocytopenia Based on High-Performance Liquid Chromatograph, Network Pharmacology, Molecular Docking and Cytokines Validation.

We explored the mechanisms and molecular targets of Ejiao Siwu Decoction (EJSW) for treating primary immune thrombocytopenia (ITP) using network pharmacology and molecular docking. Active compounds of EJSW were identified by high-performance liquid chromatography-diode array detector (HPLC-DAD) and high-performance liquid chromatography-mass spectrometry (HPLC-MS) and their targets were obtained from HERB and SwissTargetPrediction, and ITP targets were obtained from Comparative Toxicogenomics Database (CTD) and GeneCards. STRING and Cytoscape were used for protein-protein interaction (PPI) network analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses by WebGestalt yielded a gene-pathway network, Autodock molecular docking was applied to screen targets and active compounds, and cytokines were detected using a cytometric bead array (CBA) human inflammation kit. We identified 14 compounds and 129 targets, and 1,726 ITP targets. RAC-alpha serine/threonine-protein kinase (AKT1), tumour necrosis factor (TNF), interleukin-6 (IL6), caspase-3 (CASP3) and tumour suppressor protein (TP53) were core targets (nodes and edges). Functional annotation identified cofactor binding and coenzyme binding, and 20 significantly enriched pathways. Active compounds of EJSW were successfully docked with ITP targets. Tumour necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1ß) were upregulated in ITP patients, vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor D (VEGF-D) were downregulated, and EJSW treatment reversed these trends. EJSW may regulate key ITP targets based on the in silico analyses, and protect vascular integrity through AGE-RAGE signalling, complement and coagulation cascades, and VEGF signalling by downregulating TNF-α, IL-1ß and other inflammatory factors.

Evaluation of the immunogenicity of auxotrophic Lactobacillus with CRISPR-Cas9D10A system-mediated chromosomal editing to express porcine rotavirus capsid protein VP4.

Porcine rotavirus (PoRV) is an important pathogen, leading to the occurrence of viral diarrhoea . As the infection displays obvious enterotropism, intestinal mucosal immunity is the significant line of defence against pathogen invasion. Moreover, as lactic acid bacteria (LAB) show acid resistance, bile salt resistance and immune regulation, it is of great significance to develop an oral vaccine. Most traditional plasmid delivery vectors use antibiotic genes as selective markers, easily leading to antibiotic accumulation. Therefore, to select a food-grade marker in genetically engineering food-grade microorganisms is vital. Based on the CRISPR-Cas9D10A system, we constructed a stable auxotrophic Lactobacillus paracasei HLJ-27 (Lactobacillus △Alr HLJ-27) strain. In addition, as many plasmids replicate in the host bacteria, resulting in internal gene deletions. In this study,we used a temperature-sensitive gene editing plasmidto insert the VP4 gene into the genome, yielding the insertion mutant strains VP4/△Alr HLJ-27, VP4/△Alr W56, and VP4/W56. This recombinant bacterium efficiently induced secretory immunoglobulin A (SIgA)-based mucosal and immunoglobulin G (IgG)-based humoral immune responses. These oral mucosal vaccines have the potential to act as an alternative to the application of antibiotics in the future and induce efficient immune responses against PEDV infection.

Dietary α-lipoic acid supplementation improves postmortem color stability of the lamb muscles through changing muscle fiber types and antioxidative status.

This study investigated the effect of dietary α-lipoic acid (600 mg/kg) supplementation on the postmortem color stability of the biceps femoris from lambs. The results showed that dietary α-lipoic acid supplementation increased a* and decreased b* and metmyoglobin (MMb) percentage of the biceps femoris with the time of storage (P < 0.05). The content of malondialdehyde (MDA) reduced with the time of storage after treatment with α-lipoic acid (P < 0.05). α-lipoic acid increased the myoglobin (Mb) content, and myosin heavy chain I (MyHC I) gene expression but decreased glycogen content, lactate dehydrogenase (LDH) activity, and MyHC IIb gene expression (P < 0.05). The T-AOC value, catalase (CAT) activity, and expression of SOD and CAT gene expression increased after α-lipoic acid treatment (P < 0.05). Therefore, dietary α-lipoic acid supplementation improved the meat color by regulating muscle fiber types and inhibited glycolysis. Moreover, α-lipoic acid maintained meat color stability by effectively inhibiting muscle oxidation via enhancing the antioxidant capacity.

[Analysis of Differential Proteins Related to Platelet Activation in Patients with Essential Thrombocythemia Based on Label-Free Quantitative Technology].

OBJECTIVE: To analysis the specific protein markers of essential thrombocythemia (ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation. METHODS: Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P＜0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P＜0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring (PRM) technology. RESULTS: A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing. CONCLUSION: Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV, GPVI, COL1A1 and GP1bα can be used as new targets related to ET platelet activation.

Analysis of Differential Proteins Related to Platelet Activation in Patients with Essential Thrombocythemia Based on Label-Free Quantitative Technology / 中国实验血液学杂志

OBJECTIVE@#To analysis the specific protein markers of essential thrombocythemia (ET) based on proteomics technology, to explore and verify the differential protein related to platelet activation.@*METHODS@#Blood samples were obtained from ET patients and healthy people and a certain protein mass spectrometry was detected using label-free quantitative technology. The proteins relative abundance increased or down-regulated by 1.3 times in the disease group compared with the control group, and the protein abundance in the two groups t test P＜0.05 were defined as differential proteins. Bioinformatics analysis of the differential proteins was performed using GO and KEGG. The difference in the average protein abundance between the two groups was analyzed by t test and P＜0.05 was considered statistically significant. Differential proteins were selected for verification by parallel reaction monitoring (PRM) technology.@*RESULTS@#A total of 140 differential proteins were found, of which 72 were up-regulated and 68 were down-regulated. KEGG enrichment showed that the differential protein expression was related to the platelet activation pathway. The differential proteins related to platelet activation were GPV, COL1A2, GP1bα, COL1A1 and GPVI. Among them, the expressions of GPV, GP1bα and GPVI were up-regulated, and the expressions of COL1A2 and COL1A1 were down-regulated. PRM verification of COL1A1, GP1bα, GPVI and GPV was consistent with LFP proteomics testing.@*CONCLUSION@#Differential proteins in ET patients are related to platelet activation pathway activation.Differential proteins such as GPV, GPVI, COL1A1 and GP1bα can be used as new targets related to ET platelet activation.

Immune Responses in Pregnant Sows Induced by Recombinant Lactobacillus johnsonii Expressing the COE Protein of Porcine Epidemic Diarrhea Virus Provide Protection for Piglets against PEDV Infection.

Porcine epidemic diarrhea (PED) induced by porcine epidemic diarrhea virus (PEDV) is an intestinal infectious disease in pigs that causes serious economic losses to the pig industry. To develop an effective oral vaccine against PEDV infection, we used a swine-origin Lactobacillus johnsonii (L. johnsonii) as an antigen delivery carrier. A recombinant strain pPG-T7g10-COE/L. johnsonii (L. johnsonii-COE) expressing COE protein (a neutralizing epitope of the viral spike protein) was generated. The immunomodulatory effect on dendritic cell in vitro and immunogenicity in pregnant sows was evaluated following oral administration. L. johnsonii-COE could activate monocyte-derived dendritic cell (MoDC) maturation and triggered cell immune responses. After oral vaccination with L. johnsonii-COE, levels of anti-PEDV-specific serum IgG, IgA, and IgM antibodies as well as mucosal secretory immunoglobulin A (SIgA) antibody were induced in pregnant sows. High levels of PEDV-specific SIgA and IgG antibodies were detected in the maternal milk, which provide effective protection for the piglets against PEDV infection. In summary, oral L. johnsonii-COE was able to efficiently activate anti-PEDV humoral and cellular immune responses, demonstrating potential as a vaccine for use in sows to provide protection of their piglets against PEDV.

Bioinformation analysis of chorismate synthase in Baphicacantus cusia and other 78 species of plants / 中国中药杂志

Chorismate synthase(CS, EC:4.2.3.5) catalyses 5-enolpyruvy-shikimate-3-phosphate to form chorismate, which is the essential enzyme for chorismate biosynthesis in organisms. The amino acid sequences of CS from 79 species of higher plants were reported in GenBank at present. 125 amino acid sequences of CS from Baphicacanthus cusia and other 78 species of plants were predicted and analyzed by using various bioinformatics software, including the composition of amino acid sequences, signal peptide, leader peptide, hydrophobic/hydrophilic, transmembrane structure, coiled-coil domain, protein secondary structure, tertiary structure and functional domains. The phylogenetic tree of CS protein family was constructed and divided into eight groups by phylogenetic analysis. The homology comparison indicated that B. cusia shared a high homology with several plants such as Sesamum indicum, Nicotiana tabacum, Solanum tuberosum and so on. The open reading frame(ORF) of all samples is about 1 300 bp, the molecular weight is about 50 kDa, the isoelectric point(pI) is 5.0-8.0 which illustrated that CS protein is slightly basic. The ORF of CS we cloned in B. cusia is 1 326 bp, the amino acid residues are 442, the molecular weight is 47 kDa and pI is 8.11. The CS in B.cusia showed obvious hydrophobicity area and hydrophilicity area, no signal peptide, and may exists transmembrane structure areas. The main secondary structures of CS protein are random coil and Alpha helix, also contain three main structural domains which are an active structural domain, a PLN02754 conserved domain and a FMN binding site. The acquired information in this study would provide certain scientific basis for further study on structure-activity relationship and structure modification of CS in plants in the future.

[Biodiversity of Lactobacilli from fermented milk products in Northern Tibet].

OBJECTIVE: In order to study the biodiversity of Lactobacilli from Northern Tibet in China, we developed enterobacterial repetitive intergenic consensus sequence polymerase chain reaction (ERIC-PCR) technique. METHODS: Seventy seven Lactobacillus strains were isolated from traditional fermented milk products which are made by herd man families in Northern Tibet of China. By ERIC-PCR technique method, using NTSYS-pc2.1 software, we investigated the biodiversity of the strains. RESULTS: The amplified bands of ERIC-PCR were clear and stable with high repetition and high polymorphisms. The cluster analysis showed that at level 0.73, 77 tested strains were clustered into 4 groups: A1: strains of Lactobacillus casei (35.06% of the strains), A2: strains of Lactobacillus fermentum (61.04% of the strains), A3: strains of Lactobacillus helveticus and A4: strains of Lactobacillus plantarum. The biodiversity of the isolates from Aland A2 groups was further investigated. A1 could be divided into 5 genotypes at the similarity of 80.0%. A2 could be divided into 6 genotypes at the similarity of 84.0%. CONCLUSION: Using ERIC-PCR technique, Lactobacilli from Northern Tibet could be distinguished at species level.

Natural gas and indoor air pollution: a comparison with coal gas and liquefied petroleum gas.

OBJECTIVE: The study was designed to compare the combustion products of coal gas, liquefied petroleum gas and natural gas in relation to indoor air pollution. METHODS: Regular pollutants including B(a)P were monitored and 1-hydroxy pyrene were tested in urine of the enrolled subjects. Radon concentrations and their changes in four seasons were also monitored in the city natural gas from its source plant and transfer stations to final users. To analyze organic components of coal gas, liquefied petroleum gas and natural gas, a high-flow sampling device specially designed was used to collect their combustion products, and semi-volatile organic compounds contained in the particles were detected by gas chromatograph-mass spectrograph (GC/MS). RESULTS: Findings in the study showed that the regular indoor air pollutants particles and CO were all above the standard in winter when heating facilities were operated in the city, but they were lowest in kitchens using natural gas; furthermore, although NO2 and CO2 were slightly higher in natural gas, B(a)P concentration was lower in this group and 1-hydroxy pyrene was lowest in urine of the subjects exposed to natural gas. Organic compounds were more complicated in coal gas and liquefied petroleum gas than in natural gas. The concentration of radon in natural gas accounted for less than 1% of its effective dose contributing to indoor air pollution in Beijing households. CONCLUSION: Compared to traditional fuels, gases are deemed as clean ones, and natural gas is shown to be cleaner than the other two gases.

Natural gas and indoor air pollution: a comparison with coal gas and liquefied petroleum gas / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>The study was designed to compare the combustion products of coal gas, liquefied petroleum gas and natural gas in relation to indoor air pollution.</p><p><b>METHODS</b>Regular pollutants including B(a)P were monitored and 1-hydroxy pyrene were tested in urine of the enrolled subjects. Radon concentrations and their changes in four seasons were also monitored in the city natural gas from its source plant and transfer stations to final users. To analyze organic components of coal gas, liquefied petroleum gas and natural gas, a high-flow sampling device specially designed was used to collect their combustion products, and semi-volatile organic compounds contained in the particles were detected by gas chromatograph-mass spectrograph (GC/MS).</p><p><b>RESULTS</b>Findings in the study showed that the regular indoor air pollutants particles and CO were all above the standard in winter when heating facilities were operated in the city, but they were lowest in kitchens using natural gas; furthermore, although NO2 and CO2 were slightly higher in natural gas, B(a)P concentration was lower in this group and 1-hydroxy pyrene was lowest in urine of the subjects exposed to natural gas. Organic compounds were more complicated in coal gas and liquefied petroleum gas than in natural gas. The concentration of radon in natural gas accounted for less than 1% of its effective dose contributing to indoor air pollution in Beijing households.</p><p><b>CONCLUSION</b>Compared to traditional fuels, gases are deemed as clean ones, and natural gas is shown to be cleaner than the other two gases.</p>