Compensatory effects of other olfactory genes after CRISPR/cas9 editing of BmOR56 in silkworm, Bombyx mori.

Bombyx mori is an oligophagous economic insect. Cis-Jasmone is one of the main substances in mulberry leaf that attract silkworm for feeding and BmOR56 is its receptor. Potential interaction ways between BmOR56 and cis-Jasmone were explored, which included some crucial amino acids such as Gln172, Val173, Ser176, Lys182, His322, and Arg345. BmOR56 was edited using CRISPR/cas9 for Qiufeng, and a homozygous knockout strain QiufengM was obtained. Compared with Qiufeng, the feeding ability of QiufengM on mulberry leaf did not change significantly, but on artificial diet decreased significantly. QiufengM also showed a dependence on the concentration of mulberry leaf powder. The result indicated that other olfactory genes had a compensatory effect on the attractance of mulberry leaf after the loss of BmOR56. Transcriptome analysis of antennae showed that many genes differentially expressed between Qiufeng and QiufengM, which involved in olfactory system, glucose metabolism, protein metabolism, amino acid metabolism, and insect hormone biosynthesis. Particularly, BmIR21, BmOR53 and BmOR27 were significantly up-regulated, which may have a compensatory effect on BmOR56 loss. In addition, detoxification mechanism was activated and may cause the passivation of feeling external signals in silkworm.

On-site monitoring of nandrolone in cattle farming samples by portable atmospheric pressure chemical ionization mass spectrometry with ambient sampling.

Nandrolone (NT) is a type of androgen anabolic steroid that is often illegally used in cattle farming, leading to unpredictable harm to human health via the food chain. In this study, a rapid detection method for NT in the samples of cattle farming was established using a portable mass spectrometer. The instrument parameters were optimized, including a thermal desorption temperature of 220 °C, a pump speed of 30 %, an APCI ionization voltage of 3900 v, and an injection volume of 6 µL. The samples of bovine urine, feed, sewage, and tissue were selected, and extracted using a solution of methanol:acetonitrile (1:1, v/v), followed by spiking a NT standard solution (1000 ng·mL-1) and ionization through the APCI ion source for detection. The results showed that NT could not be detected in beef and feed due to the complexity of the matrix, while clear signals of NT ions were observed in bovine urine and sewage samples, with LODs of 1000 and 100 ng·mL-1, respectively. Furthermore, quantitative analysis was attempted, and a good linear relationship (R2 = 0.9952) was observed for NT in sewage within the range of 100 to 1000 ng·mL-1. At spiked levels of 100, 500, 1000 and 2000 ng mL-1, the recovery rates ranged from 74.3 % to 92.8 %, with a relative standard deviation (n = 6) of less than 15 %. In conclusion, this detection method offers the advantages of simplicity, rapidity, strong timeliness, and specificity, making it suitable for on-site detection. It can be used for qualitative screening of nandrolone in bovine urine and quantitative analysis of nandrolone in sewage.

Bombyx mori triose-phosphate transporter protein inhibits Bombyx mori nucleopolyhedrovirus infection by reducing the cell glycolysis pathway.

Bombyx mori triose-phosphate transporter protein (BmTPT) is a member of the solute carrier (SLC) family. Its main function is to transport triose phosphate between intracellular and extracellular. In this study, BmTPT was cloned and characterised from the fat body of the silkworm Bombyx mori, resulting in an open reading frame (ORF) with a full length of 936 bp, which can encode 311 amino acid residues and has eight transmembrane structural domains. BmTPT was distributed throughout the cell and deposited the most in the nucleus, and is expressed in all tissues of Bombyx mori. Bombyx mori nucleopolyhedrovirus (BmNPV) infection significantly up-regulated BmTPT expression in immune tissue fat bodies. In addition, overexpression of BmTPT significantly inhibited BmNPV infection and markedly reduced the expression of enzymes related to the cellular glycolytic pathway; on the contrary, down-regulation of BmTPT expression by RNA interference resulted in robust replication of BmNPV and a significant increase in the expression of enzymes related to the cellular glycolytic pathway. This is the first report that BmTPT has antiviral effect in silkworm, and also could result in a lack of energy and raw materials for BmNPV replication and infection through down-regulation of the cellular glycolytic pathway.

BmC/EBPZ gene is essential for the larval growth and development of silkworm, Bombyx mori.

The genetic male sterile line (GMS) of the silkworm Bombyx mori is a recessive mutant that is naturally mutated from the wild-type 898WB strain. One of the major characteristics of the GMS mutant is its small larvae. Through positional cloning, candidate genes for the GMS mutant were located in a region approximately 800.5 kb long on the 24th linkage group of the silkworm. One of the genes was Bombyx mori CCAAT/enhancer-binding protein zeta (BmC/EBPZ), which is a member of the basic region-leucine zipper transcription factor family. Compared with the wild-type 898WB strain, the GMS mutant features a 9 bp insertion in the 3'end of open reading frame sequence of BmC/EBPZ gene. Moreover, the high expression level of the BmC/EBPZ gene in the testis suggests that the gene is involved in the regulation of reproduction-related genes. Using the CRISPR/Cas9-mediated knockout system, we found that the BmC/EBPZ knockout strains had the same phenotypes as the GMS mutant, that is, the larvae were small. However, the larvae of BmC/EBPZ knockout strains died during the development of the third instar. Therefore, the BmC/EBPZ gene was identified as the major gene responsible for GMS mutation.

Evaluation of Lipid Oxidation Characteristics in Salmon after Simulation of Cold Chain Interruption Using Rapid Evaporation Ionization Mass Spectrometry.

Temperature fluctuations occurring during the cold chain logistics of salmon contribute to lipid oxidation. This study aimed to simulate cold chain interruption through freeze-thaw operations and evaluate the lipidomics data from salmon samples subjected to different numbers of freeze-thaw cycles by using rapid evaporative ionization mass spectrometry (REIMS) combined with an intelligent surgical knife (iKnife). The results indicated significant differences in the relative abundance of characteristic ions among the samples (p < 0.05). A total of 34 ions with variable importance for the projection values ≥1 were identified as potential biomarkers, including m/z 719.4233 ([PCC36:5-NH(CH3)3]-), m/z 337.3134 ([FAC22:1]-), m/z 720.4666 ([PEC35:6-H]-), m/z 309.2780 ([FAC20:1]-), m/z 777.4985 ([PCC40:4-NH(CH3)3]-), m/z 745.4421 ([PCC38:6-NH(CH3)3]-/[PEC38:6-NH3]-), m/z 747.4665 ([PCC38:5-NH(CH3)3]-/[PEC38:5-NH3]-), etc. The degree of lipid oxidation was found to be associated with the number of freeze-thaw cycles, exhibiting the most significant alterations in the relative abundance of lipid ions in the 8T samples. Additionally, sensory evaluation by the CIE-L*a*b* method and volatile analysis by headspace solid-phase microextraction gas chromatography-mass spectrometry demonstrated significant differences (p < 0.05) in color and odor among the salmon samples, with a correlation to the number of freeze-thaw cycles. The primary compounds responsible for alterations in salmon odor were aldehydes with lower odor thresholds. In summary, the iKnife-REIMS method accurately differentiated salmon muscle tissues based on varying levels of lipid oxidation, thus expanding the application of REIMS.

Ras3 in Bombyx mori with antiviral function against B. mori nucleopolyhedrovirus.

Bombyx mori ras protein3 (BmRas3) is a small molecular protein in the GTPase superfamily, which has the activity of binding guanosine nucleotides and GTP enzymes. It acts as a molecular switch by coupling extracellular signal to different cellular response through the conversion between Ras-GTP conformation and Ras-GDP conformation, thus regulating signal pathways responsible for cell growth, migration, adhesion, survival and differentiation. However, few studies have been done on Ras3 in silkworm, and its function and mechanism are unclear. In this study, we found that the overexpression of BmRas3 inhibited the infection of BmNPV(B. mori nucleopolyhedrovirus), while knockdown of BmRas3 could promote the infection of BmNPV. In addition, after the BmRas3 in silkworm larvae was knockdown, the anti-BmNPV ability of silkworm decreased and the survival rate of silkworm was affected. Additionly in the cells with BmRas3 overexpression, the transcription level of BmMapkk6 、BmP38、BmJNK、BmERK1/2 and BmERK5 were significantly increased after BmNPV infection, and the transcript levels of BmMapkk6、BmP38、BmJNK、BmERK1/2 and BmERK5 were also inhibited to varying degrees This is the first report on the antiviral effect of BmRas3 in silkworm, which provides a new direction for further study on the anti-BmNPV mechanism of silkworm and screening and cultivation of anti-BmNPV silkworm strain.

Genetic and transcriptome analysis of the smb mutant in Bombyx mori.

More than 600 mutations have been discovered in the history of silkworm domestication. It is important to study the formation mechanism of these mutations to further understand the life and development process of silkworms and agricultural pest control. The silkworm mutant smb was isolated from silkworm strain NCV, and transcriptome analysis was performed on the silkworm mutant. 796 differentially expressed genes (DEGs) were detected at 48 h of the second instar stage with 669 genes significantly upregulated and 127 genes significantly downregulated. During the GO enrichment analysis, it was found that the enrichment of biological processes was mainly concentrated in proteolysis, carbohydrate metabolism, aminoglycan metabolism, organic substance metabolism, protein metabolism and so on. Based on the analysis of KEGG pathways, it revealed that the pathways enriched in lysosomes, AMPK signaling, fatty acid metabolism, PPAR signaling, galactose metabolism, and protein digestion and absorption were the most significant. Through these most significantly enriched GO terms and KEGG pathways, DEGs consistent with the phenotypic characteristics of the smb mutant were identified, including small body size, slow development, and successive death after the fourth instar. These results provided experimental evidence for the potential formation mechanism of smb mutants.

Determination of Pentachlorophenol in Seafood Samples from Zhejiang Province Using Pass-Through SPE-UPLC-MS/MS: Occurrence and Human Dietary Exposure Risk.

Pentachlorophenol (PCP) has attracted wide attention due to its high toxicity, persistence, and bioaccumulation. In this study, a sensitive UPLC-MS/MS method for the determination of PCP in seafood samples was developed and validated. The samples were ultrasonic extracted with acetonitrile containing 1% acetic acid-acetonitrile and followed by using a pass-through solid-phase extraction (SPE) cleanup on Captiva EMR-Lipid cartridges. The linearity of this method ranged from 1 to 1000 µg/L, with regression coefficients of >0.99. The detection limit and quantitation limit were 0.5 µg/kg and 1.0 µg/kg, respectively. The recoveries in different types of seafood samples ranged from 86.4% to 102.5%, and the intra-day and inter-day relative standard deviations (RSDs) were 3.7% to 11.2% and 2.9% to 12.1%, respectively (n = 6). Finally, the method has been successfully utilized for the screening of PCP in 760 seafood samples from Zhejiang Province. PCP was detected in 5.8% of all seafood samples, with the largest portion of detections found in shellfish, accounting for approximately 60% of the total. The average concentrations detected ranged from 1.08 to 21.49 µg/kg. The non-carcinogenic risk indices for adults and children who consume PCP ranged from 10-4 to 10-3 magnitudes. All of these indices stayed significantly below 1, implying that the health risk from PCP in marine organisms to humans is minimal.

Ultrasound and clinicopathological characteristics of breast cancer for predicting axillary lymph node metastasis.

OBJECTIVES: The goal of this study was to assess the clinicopathological and ultrasound (US) features of breast cancer for predicting the risk of axillary lymph node metastasis. METHODS: Patients with breast cancer were included in this retrospective, monocentric, observational study. Their preoperative ultrasound features, clinical data, laboratory results and postoperative pathologic results and immunophenotyping were collected. The association of these factors of breast cancer with axillary lymph node metastasis was evaluated by univariate and multivariate analysis. RESULTS: In this study, 471 patients diagnosed with breast cancer at the First Affiliated Hospital of Xi'an Jiaotong University between July 2016 and September 2019 were collected, with a total of 471 nodules, of which 231(49.0%) had axillary lymph node metastasis, and 240(51.0%) did not. The parameters of hyperechoic halo, posterior acoustic decrease, microcalcification, carcinogenic embryonic antigen (CEA), cancer antigen-153 (CA153), CK5/6 (+), Ki67 (≥40%), AR (+) and histological grade (grade II and grade III) were significantly and independently associated with axillary lymph node metastasis (p < 0.05 for all). CONCLUSIONS: The combination of ultrasound features, tumor markers, pathology, and immunohistochemistry can predict axillary lymph node metastasis in breast cancer patients.

Transcriptome analysis of immune-related genes of Asian corn borer (Ostrinia furnacalis [Guenée]) after oral bacterial infection.

The Asian corn borer (Ostrinia furnacalis) is an important agricultural pest causing serious damage to economic crops, such as corn and sorghum. The gut is the first line of defense against pathogens that enter through the mouth. Staphylococcus aureus was used to infect the O. furnacalis midgut to understand the midgut immune mechanism against exogenous pathogens to provide new ideas and methods for the prevention and control of O. furnacalis. A sequencing platform was used for genome assembly and gene expression. The unigene sequences were annotated and functionally classified by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Significant differences were found in the induced expression profiles before and after infection. Some differentially expressed genes have important relations with lipid metabolism and immune mechanism, suggesting that they play an important role in the innate immune response of O. furnacalis. Furthermore, quantitative real-time polymerase chain reaction assay was used to identify the key genes involved in the signaling pathway, and the expression patterns of these key genes were confirmed. The results could help study the innate immune system of lepidopteran insects and provide theoretical support for the control of related pests and the protection of beneficial insects.

Bombyx mori Tetraspanin A (BmTsp.A) is a facilitator in BmNPV invasion by regulating apoptosis.

BmTsp.A (Bombyx mori Tetraspanin A) is one of the four transmembrane proteins which are capable to regulate multiple aspects of the immune response and are involved in various stages of viral invasion of the hosts. This study focused on the sequence features, analysis of expression pattern, as well as the effect of BmTsp.A on BmNPV (Bombyx mori nucleopolyhedrovirus) infection in the apoptotic pathway. BmTsp.A features the typical tetraspanins family, including four transmembrane domains and a major large extracellular loop domain. It is highly expressed specifically in the malpighian tubes, and its expression is increased by BmNPV induction for 48 h and 72 h. Overexpression and RNAi (RNA interference) mediated by siRNA reveal that BmTsp.A can promote the infection and replication of the virus. In addition, the overexpression of BmTsp.A regulates BmNPV-induced apoptosis, leading to changes in the expression of apoptosis-related genes and thus affecting viral proliferation. When subjected to stimulation by BmNPV infection, on the one hand, BmTsp.A inhibits Bmp53 through a Caspase-dependent pathway, which consequently promotes the expression of Bmbuffy, thereby activating BmICE to inhibit apoptosis and causing the promotion of viral proliferation. On the other hand, BmTsp.A inhibits the expression of BmPTEN and BmPkc through the phosphatidylinositol 3 kinase (PI3K)/protein kinaseB(AKT) signaling pathway, thus affecting the regulation of apoptosis. To summarize, our results demonstrate that BmTsp.A promotes viral infection and replication by inhibiting apoptosis, which is fundamental for understanding the pathogenesis of BmNPV and the immune defense mechanism of silkworm.

Transcriptome analysis of anorexic and preferred silkworms (Bombyx mori) on artificial diet.

The silkworm, Bombyx mori, is an important oligophagous economic insect and feeding habits of different silkworm varieties to artificial diet are different. Research on the mechanisms of feeding habits on artificial diet, and breeding of silkworm varieties adapted on artificial diet, which is a necessary condition for industrial silkworm rearing, is currently lacking. For an artificial diet, Xin was anorexic, whereas Haoyue A showed a strong appetite. When the two varieties were crossed, the F1 generation showed a poor appetite for the artificial diet and had a setae dispersion rate of <50 %. However, the F2 generation, self-bred progeny of F1, had a good appetite for the artificial diet, with a setae dispersion rate of 70 %. Herein, transcriptome analysis was conducted on the F2 generation, comparing individuals with anorexic and preferred feeding habits, and 2188 differential genes were identified, with 1524 genes up-regulated and 934 genes down-regulated. Several genes were identified to contribute to feeding habits, such as genes involved olfactory system, energy supply, and cell proliferation and differentiation. GO enrichment revealed a large number of DEGs related to behavior, growth, signaling, developmental process, response to stimulation, and other pathways. Furthermore, proteins closely related to feeding were expressed differently. Some DEGs were selected for qRT-PCR, and results indicated the reliability of the DEG results. The DEGs between individuals with anorexic and preferred feeding habits were screened by RNA-Seq technology, which provides a reliable reference to study molecule mechanisms of feeding habits on artificial diet.

Remnant Cholesterol and Common Carotid Artery Intima-Media Thickness in Community Population with Normal Low-Density Lipoprotein Cholesterol.

INTRODUCTION: Remnant cholesterol is a risk factor for cardiovascular disease, especially when low-density lipoprotein cholesterol (LDL-C) levels are normal. However, there are few studies on the relationship between remnant cholesterol and subclinical atherosclerosis. Common carotid artery intima-media thickness (cIMT) is an imaging marker of subclinical atherosclerosis. This study aimed to investigate the relationship between remnant cholesterol and cIMT in a community population with normal LDL-C. METHODS: This study is a retrospective analysis; 1,101 community population with available carotid artery imaging and fasting lipid data with LDL-C <4.1 mmol/L were included in this analysis. Remnant cholesterol was calculated as total cholesterol minus LDL-C minus high-density lipoprotein cholesterol. Abnormal cIMT was defined as maximum cIMT value ≥1 mm. Logistic regression was used to assess the relationships between remnant cholesterol levels and abnormal cIMT. RESULTS: As the remnant cholesterol level increased from the lowest to the highest quartile, the rate of abnormal cIMT increased from 24.5% to 38.6% (p trend <0.001) in the community population with normal LDL-C level. In the unadjusted model, the odds ratios (ORs, 95% confidence intervals) in the highest quartile group were 1.937 (1.338-2.803) for abnormal cIMT compared with the lowest quartile. The multivariable-adjusted ORs (95% confidence intervals) for the highest versus lowest quartile of remnant cholesterol were 2.132 (1.420-3.202) for abnormal cIMT. CONCLUSION: Elevated fasting remnant cholesterol levels were positively associated with abnormal cIMT in community population with normal LDL-C levels. Remnant cholesterol may be an important indicator of risk stratification in community population with normal LDL-C level.

Correction: Mei et al. Bombyx mori C-Type Lectin (BmIML-2) Inhibits the Proliferation of B. mori Nucleopolyhedrovirus (BmNPV) through Involvement in Apoptosis. Int. J. Mol. Sci. 2022, 23, 8369.

In the original publication [...].

In Situ Construction of MnO2-Co3O4 Nanosheet Heterojunctions on Co@NCNT Surfaces for Oxygen Evolution.

Electrocatalytic water splitting is still circuitous and controversial because of the lack of highly active electrocatalysts to decrease the overpotential. Herein, we report a feasible method for constructing heterojunctions of MnO2-Co3O4 nanosheets on Co@NCNT support surfaces (MnO2-Co3O4/Co@NCNT) by spontaneous redox reactions. Experimental results indicate that Co embedded in Co@NCNT can be used as the carbon support and anchoring sites for heterojunctions, thus exposing a large number of active sites, adjusting the surface electronic structure, changing the OER rate-determining step of the catalyst, and reducing the reaction energy barrier. Besides, the in situ formation of MnO2-Co3O4 nanosheets on Co@NCNT inhibits the loss and aggregation of the catalyst, leading to robust structural stability. Therefore, the synergistic effects of these factors provide multi-functional active sites to enhance the intrinsic activity and achieve maximum catalytic performances. To deliver a current density of 10 mA cm-2, the catalyst of MnO2-Co3O4/Co@NCNT achieves an overpotential (η) of 303 mV in 1.0 M KOH media for OER. This simple redox strategy can be easily extended to prepare other ultrathin transition-metal oxide heterojunctions, which could be applied not only for water splitting but also for other energy conversion and storage technologies.

Chain-locked precursor ion scanning based HPLC-MS/MS for in-depth molecular analysis of lipase-catalyzed transesterification of structured phospholipids containing ω-3 fatty acyl chains.

Lipase-catalyzed transesterification of structured phospholipids (sPLs) is a hot topic, but the structural variation of the fatty acyl chains in intact phospholipids at the molecular level remains unclear to date. The present study explored the detailed characteristics of synthesized phospholipids through high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in precursor ion scan mode. The optimal conditions were in-depth inspected and determined for the reaction system, including phospholipase A1 as catalyst, 15% lipase loading, and 1% water content. The sPLs enriched with EPA/DHA were structurally and quantitatively characterized by focusing on the fragments of m/z 301.6 (eicosapentaenoic acid, EPA) and m/z 327.6 (docosahexaenoic acid, DHA), and the results were statistically analyzed using partial least squares discriminant analysis and clustered heatmap hierarchical clustering analysis. PC 38:6 (18:1/20:5), PC 38:7 (18:2/20:5), PC o-40:6 (o-18:0/22:6), and PE 40:8 (18:2/22:6) etc. were revealed as the main variables that were active in the reaction.

Study on the physicochemical indexes, nutritional quality, and flavor compounds of Trichiurus lepturus from three representative origins for geographical traceability.

Trichiurus lepturus (hairtail) is an important economic component of China's marine fishing industry. However, due to the difficulty in identifying the appearance of hairtail from different geographical distributions, hairtails with geographical indication trademarks were imitated by general varieties. In this study, the texture characteristics, color, basic nutrients, amino acids, mineral, fatty acids, and volatile flavor substances were used as indicators for multivariate statistical analysis to determine whether three origins of hairtails from the habitats of Zhoushan (East China Sea, T.Z), Hainan (South China Sea, T.N), and Qingdao (Yellow Sea, T.Q) in the market could be distinguished. The findings revealed that there were significant differences in amino acids composition, mineral composition, fatty acid composition in lipids, and volatile flavor substances among the hairtails of three origins (P < 0.05), but no differences in color, texture, protein content. T.Z had moisture, crude fat, essential amino acids (EAA), flavor amino acids (FAA), unsaturated fatty acids (UFA), and docosahexaenoic acids and dicosapentaenoic acids (ΣEPA + DHA) contents of 74.33, 5.4%, 58.25 mg⋅g-1, 46.20 mg⋅g-1, 66.84 and 19.38%, respectively, and the contents of volatile alcohols, aldehydes and ketones were 7.44, 5.30, and 5.38%, respectively. T.N contains moisture, crude fat, EAA, FAA, UFA and ΣEPA + DHA as 77.69, 2.38%, 64.76 mg⋅g-1, 52.44 mg⋅g-1, 65.52 and 29.45%, respectively, and the contents of volatile alcohols, aldehydes and ketones as 3.21, 8.92, and 10.98%, respectively. T.Q had the contents of moisture, crude fat, EAA, FAA, UFA, and ΣEPA + DHA 79.69, 1.43%, 60.9 mg⋅g-1, and 49.42 mg⋅g-1, respectively. The contents of unsaturated fatty acid and ΣEPA + DHA were 63.75 and 26.12%, respectively, while the volatile alcohols, aldehydes, and ketones were 5.14, 5.99, and 7.85%, respectively. Partial least squares-discriminant analysis (PLS-DA) multivariate statistical analysis showed that volatile flavor compounds could be used as the most ideal indicators for tracing the source of hairtail. In conclusion, the findings of this study can distinguish the three hairtail origins using some basic indicators, providing ideas for hairtail geographical identification.

Immunoenhancing Effects of Cyclina sinensis Pentadecapeptide through Modulation of Signaling Pathways in Mice with Cyclophosphamide-Induced Immunosuppression.

Our study aimed to investigate the immune-enhancing mechanism of the pentadecapeptide (RVAPEEHPVEGRYLV) from Cyclina sinensis (SCSP) in a cyclophosphamide (CTX)-induced murine model of immunosuppression. Our results showed that SCSP treatment significantly increased mouse body weight, immune organ indices, and the production of serum IL-6, IL-1ß, and tumor necrosis factor (TNF)-α in CTX-treated mice. In addition, SCSP treatment enhanced the proliferation of splenic lymphocytes and peritoneal macrophages, as well as phagocytosis of the latter in a dose-dependent manner. Moreover, SCSP elevated the phosphorylation levels of p38, ERK, JNK, PI3K and Akt, and up-regulated IKKα, IKKß, p50 NF-κB and p65 NF-κB protein levels, while down-regulating IκBα protein levels. Our results indicate that SCSP has immune-enhancing activities, and that it can activate the MAPK/NF-κB and PI3K/Akt pathways to enhance immunity in CTX-induced immunosuppressed mice.

Peptidoglycan recognition protein 6 (PGRP6) from Asian corn borer, Ostrinia furnacalis (Guenée) serve as a pattern recognition receptor in innate immune response.

Peptidoglycan recognition proteins (PGRPs) recognize invading microbes via detecting peptidoglycans from microbial cell walls. PGRPs are highly conserved from insects to vertebrates and all play roles during the immune defensive response. Ten putative PGRPs have been identified through transcriptome analysis in the Asian corn borer, Ostrinia furnacalis (Guenée). Whereas, the biochemical functions of most of them have not yet been elucidated. In this study, we found PGRP6 messenger RNA exhibited extremely high expression levels in the midgut, and its transcript level increased dramatically upon bacterial infection. Moreover, the enzyme-linked immunosorbent assay indicated recombinant PGRP6 exhibited a strong binding affinity to peptidoglycans from Micrococcus luteus and Bacillus subtilis, which could agglutinate M. luteus and yeast Pichia pastoris. Additionally, we demonstrated that PGRP6 was involved in the pathway of antimicrobial peptides synthesis, but could not enhance encapsulation and melanization of hemocytes. Overall, our results indicated that O. furnacalis PGRP6 serves as a pattern recognition receptor and detects peptidoglycans from microbes to initiate the immune response.

Bombyx mori C-Type Lectin (BmIML-2) Inhibits the Proliferation of B. mori Nucleopolyhedrovirus (BmNPV) through Involvement in Apoptosis.

C-type lectins (CTLs) are widely distributed in mammals, insects, and plants, which act as pattern recognition receptors (PRRs) to recognize pathogens and initiate immune responses. In this study, we identified a C-type lectin gene called BmIML-2 from the silkworm Bombyx mori. Its open reading frame (ORF) encodes 314 amino acids, which contain dual tandem C-type lectin-like domain (CTLD). BmIML-2 is highly expressed in the fat body and is significantly induced at 24 h after BmNPV infection. Moreover, overexpression of BmIML-2 dramatically inhibited the proliferation of BmNPV, and knockdown assay via siRNA further validated the inhibition of BmIML-2 on viral proliferation. In addition, transcript level detection of apoptosis-related genes and observation of apoptosis bodies implied that overexpression of BmIML-2 promoted BmNPV-induced apoptosis. Immunofluorescence analysis indicated that BmIML-2 distributed throughout the cytoplasm and was slightly concentrated in the cell membrane. Taken together, our results suggest that BmIML-2 could inhibit in the proliferation of BmNPV by facilitating cell apoptosis.