Affinity-Driven Site-Specific High Mannose Modification Determines the Structural Polymerization and Function of Tetrameric IgM in a Primitive Vertebrate.

Teleost tetramer IgM is the predominant Ig in the immune system and plays essential roles in host defense against microbial infection. Due to variable disulfide polymerization of the monomeric subunits, tetrameric IgM possesses considerable structural diversity. Previous work indicated that the teleost IgM H chain was fully occupied with complex-type N-glycans. However, after challenge with trinitrophenyl (TNP) Ag, the complex N-glycans in the Asn-509 site of Oreochromis niloticus IgM H chain transformed into high mannose. This study, therefore, was conducted to examine the functional roles of the affinity-related high-mannose modification in tilapia IgM. The TNP-specific IgM Ab affinity maturation was revealed in tilapia over the response. A positive correlation between TNP-specific IgM affinity and its disulfide polymerization level of isomeric structure was demonstrated. Mass spectrometric analysis indicated that the relationship between IgM affinity and disulfide polymerization was associated with the Asn-509 site-specific high-mannose modification. Furthermore, the increase of high mannose content promoted the combination of IgM and mannose receptor (MR) on the surface of phagocytes. Moreover, the increased interaction of IgM and MR amplified the phagocytic ability of phagocytes to Streptococcus agalactiae. To our knowledge, this study demonstrates that site-specific high-mannose modification associates with IgM Ab affinity and its structural disulfide polymerization and amplifies the phagocytosis of phagocytes by the combination of IgM and MR. The present study provides evidence for understanding the association of IgM structure and function during the evolution of the immune system.

Non-Hermitian guided modes and exceptional points using loss-free negative-index materials.

We analyze the guided modes in coupled waveguides made of negative-index materials without gain or loss. We show that it supports non-Hermitian phenomenon on the existence of guided mode versus geometric parameters of the structure. The non-Hermitian effect is different from parity-time (P T) symmetry, and can be explained by a simple coupled-mode theory with an anti-P T symmetry. The existence of exceptional points and slow-light effect are discussed. This work highlights the potential of loss-free negative-index materials in the study of non-Hermitian optics.

Characteristics of Allergen Components of Dermatophagoides pteronyssinus in Shrimp and Mite Co-Sensitized Patients in Southern China.

INTRODUCTION: Dermatophagoides pteronyssinus (Dp) and shrimp are common air allergens and allergenic food sources, respectively, in southern China. This study aimed to analyze the specific immunoglobulin E (sIgE) characteristics and relationships of Dp components among co-sensitized patients with Dp and shrimp. MATERIALS AND METHODS: Serum samples were collected from 112 patients with Dp sensitization (61 with shrimp sensitization and 51 without) from southern China. The sIgE concentrations of Dp and shrimp crude extracts were determined by ImmunoCAP, and the sIgE of Dp allergen components (Der p 1, Der p 2, Der p 5, Der p 7, Der p 10, Der p 21, and Der p 23) was detected by protein chip. RESULTS: Overall, in the Dp-allergic patients, Der p 1 had the highest positive rate (72.3%), followed by Der p 2 (65.2%), Der p 23 (46.4%), Der p 7 (32.14%), Der p 21 (29.46%), Der p 5 (22.32%), and Der p 10 (17.86%). Compared with that in the shrimp nonsensitized group, the positive rate of sIgE for Der p 10 (27.87% vs. 5.88%, p = 0.002) in the shrimp sensitization group was significantly higher; however, the positive rate of sIgE for Der p 7 (22.95% vs. 43.14%, p = 0.023) was significantly lower. Moreover, the concentration of sIgE for Der p 10 increased statistically in the shrimp-sensitized group. The correlation analysis also showed that shrimp sensitization was significantly correlated with Der p 10. CONCLUSION: Among patients with Dp sensitization, Der p 1 had the highest positive rate, followed by Der p 2 and Der p 23. Meanwhile, Der p 10 may play an important role in patients with shrimp sensitization, while Der p 7 may be the meaningful allergen component in patients with Dp sensitization alone. In general, component-resolved diagnosis technology in clinical practice can effectively guide patients with polysensitization to avoid allergic substances.

Diagnostic Strategy and Prognostic Factors for Ectopic Adrenocorticotropic Hormone Syndrome: A Retrospective Study of 64 Cases.

INTRODUCTION: Ectopic adrenocorticotropic hormone (ACTH) syndrome (EAS) is a serious life-threatening disease. Tumor localization is crucial in EAS management. This underscores the importance of evaluating imaging methods and prognostic factors to provide a clear basis for patient diagnosis and management. OBJECTIVE: The aim of this study was to investigate imaging methods and analyze the relevant prognostic factors for EAS. METHODS: The retrospective study followed 64 cases of EAS diagnosed between 1992 and 2020. Clinical features, biochemical analysis, and imaging studies were collected, and survival data were followed up and analyzed. RESULTS: Of 64 patients, 41% were female with a mean (±SD) age at diagnosis of 47 ± 16 years. Computed tomography (CT), 18-F fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)-CT, and octreotide scintigraphy had similar sensitivity in localizing ectopic ACTH-secreting tumors. However, in cases with negative imaging on CT, both of 18F-FDG PET-CT and octreotide scintigraphy further localized 25% tumors. The combination of all three modalities failed to further increase the sensitivity. Patients with thymic tumors survived longer than those with pulmonary or pancreatic tumors (p = 0.013 and 0.047, respectively). Multivariate analyses showed that hypokalemia (p = 0.004) and treatment modality (p = 0.048) were independent prognostic factors. The optimal serum potassium cutoff based on maximum log-rank statistics (p = 0.012) was 2.90 mmol/L. CONCLUSION: CT is the first choice for tumor localization in EAS. CT in combination with a nuclear medicine or molecular imaging modality is necessary for further identification of an ectopic source. Serum potassium <2.90 mmol/L is associated with shorter overall survival, and tumor resection plays the most important role in the survival improvement.

METTL3-m6A-Rubicon axis inhibits autophagy in nonalcoholic fatty liver disease.

N6-methyladenosine (m6A) mRNA modification plays critical roles in various biological events and is involved in multiple complex diseases. However, the role of m6A modification in autophagy in nonalcoholic fatty liver disease (NAFLD) remains largely unknown. Here, we report that m6A modification was increased in livers of NAFLD mouse models and in free fatty acid (FFA)-treated hepatocytes, and the abnormal m6A modification was attributed to the upregulation of methyltransferase like 3 (METTL3) induced by lipotoxicity. Knockdown of METTL3 promoted hepatic autophagic flux and clearance of lipid droplets (LDs), while overexpression of METTL3 inhibited these processes. Mechanistically, METTL3 directly bound to Rubicon mRNA and mediated the m6A modification, while YTH N6-methyladenosine RNA binding protein 1 (YTHDF1), as a partner of METTL3, interacted with the m6A-marked Rubicon mRNA and promoted its stability. Subsequently, RUBICON inhibited autophagosome-lysosome fusion and further blocked clearance of LDs. Taken together, our results showed a critical role of METTL3 and YTHDF1 in regulating lipid metabolism via the autophagy pathway and provided a novel insight into m6A mRNA methylation in NAFLD.

The relationship of D. pteronyssinus allergic component sIgE and sIgG4 in house dust mite allergic rhinitis or/and allergic asthma patients.

Objective: House-dust mite sensitization is an important cause of allergic asthma and/or rhinitis in southern China. This study aimed to analyze the immune effect and relationship between the Dermatophagoides pteronyssinus components specific immunoglobulin E (sIgE) and sIgG4. Methods: The serum levels of sIgE and sIgG4 to D. pteronyssinus allergen components Der p 1, 2, 3, 5, 7, 10, and 23 were detected in 112 patients with allergic rhinitis (AR) and/or allergic asthma (AA). Results: Overall, Der p 1 had the highest positive rate of sIgE (72.3%), followed by Der p 2 (65.2%) and Der p 23 (46.4%). Meanwhile, the highest positive rates of sIgG4 were for Der p 2 (47.3%), Der p 1 (33.0%), and Der p 23 (25.0%). The patients with AR and AA had a higher positive rate (43.4%) of sIgG4 than that in the patients with AR (42.4%) and the patients with AA (20.4%; p = 0.043). In patients with AR, the positive rate of sIgE in Der p 1 (84.8%) was higher than that in sIgG4 (42.4%; p = 0.037), but the positive rate of sIgG4 in Der p 10 (21.2%) was higher than that in sIgE (18.2%; p < 0.001). Most of the patients were positive for sIgE and sIgG4 of Der p 2 and Der p 10 at the same time. However, positive results for sIgE alone were just found in Der p 7 and Der p 21. Optimal scale analysis showed that Der p 2, Der p 7, and Der p 21 sIgG4 were closely related to AR and AA (Cronbach α = 0.917). Conclusion: Herein, the D. pteronyssinus allergen components showed different characteristics among the patients with AR, patients with AA, and patients with AR and AA in southern China. Thus, sIgG4 may be play an important role in allergic reactions.

Resistance risk and resistance-related point mutations in cytochrome b of florylpicoxamid in Colletotrichum scovillei.

Anthracnose caused by Colletotrichum scovillei is one of the most destructive diseases of chili worldwide. Florylpicoxamid is a new quinone inside inhibitor (QiI) fungicide, which shows intensively inhibitory activity against C. scovillei. Currently, florylpicoxamid is in the registration process to control chili anthracnose in China. This study investigated the risk of resistance and resistance genetic mechanism of C. scovillei to florylpicoxamid. Baseline sensitivity of 141C. scovillei isolates to florylpicoxamid was established with an average EC50 value of 0.2328 ± 0.0876 µg/mL. A total of seven stable florylpicoxamid-resistant mutants were obtained with resistance factors ranging from 41 to 276. The mutants showed similar or weaker traits in mycelial growth, sporulation, conidial germination and pathogenicity than their parental isolates. Generally, the resistance risk of C. scovillei to florylpicoxamid would be moderate. In addition, there was no cross-resistance between florylpicoxamid and the commercially available fungicides tested. A37V and S207L mutations in the cytochrome b protein were detected in four high-resistance and three moderate-resistance mutants, respectively, of which, S207L is a new mutation. Molecular docking showed that the two mutations conferred different resistance levels to florylpicoxamid. These results provide a new perspective for QiI fungicide-resistance mechanism and may help in the reasonable use of florylpicoxamid against chili anthracnose in the future.

Lipotoxicity-induced circGlis3 impairs beta cell function and is transmitted by exosomes to promote islet endothelial cell dysfunction.

AIMS/HYPOTHESIS: Lipotoxicity constitutes the major driving force for type 2 diabetes. Circular RNAs (circRNAs) play important roles in regulating beta cell function and exosomes are essential mediators of intercellular communication. The role of exosomal circRNAs in type 2 diabetes remains largely unknown. We aimed to examine whether lipotoxicity induces dysregulation of circRNAs in beta cell-derived exosomes and to determine the contribution of exosomal circRNAs to the development of type 2 diabetes. METHODS: Exosomes were extracted from MIN6 cells treated with palmitate or BSA, and RNA sequencing was performed. CircGlis3 (Gli-similar 3) expression level was validated by qPCR. The impact of circGlis3 on beta cell function and the deleterious effects of exosomal circGlis3 on islet endothelial cells (islet ECs) were investigated in vitro and in vivo in human and mouse models by gain or loss of function assays. The molecular mechanism of circGlis3 was explored by RNA pull-down and immunoprecipitation assays. RESULTS: Beta cell-derived exosomal circGlis3 was significantly upregulated under lipotoxic conditions, and exosomal circGlis3 levels were also elevated in the serum of mouse models of diabetes and participants with type 2 diabetes. CircGlis3 participated in lipotoxicity-induced beta cell dysfunction in vitro and in vivo. Moreover, beta cell-derived exosomal circGlis3 could be transferred to islet ECs and reduce the cell viability, cell migration and angiogenesis of islet ECs. Mechanistically, circGlis3 promoted the degradation of glucocorticoid modulatory element-binding protein 1 (GMEB1) by facilitating the interaction between GMEB1 and mindbomb E3 ubiquitin protein ligase 2 (MIB2), thus suppressing the phosphorylation of heat shock protein 27 (HSP27). CONCLUSIONS/INTERPRETATION: Our study points to the involvement of circGlis3 in diabetes development, and exosomal circGlis3 transfer as a communication mode between beta cells and islet ECs, suggesting that circGlis3 might be a potential biomarker and therapeutic target for type 2 diabetes. DATA AVAILABILITY: The RNA-sequencing data have been deposited in the NCBI Sequence Read Archive (SRA) database, with accession number PRJNA689673. Mass spectrometry data are available via ProteomeXchange with identifier PXD024693.

Knockdown of lncRNA BDNF-AS inhibited the progression of multiple myeloma by targeting the miR-125a/b-5p-BCL2 axis.

PURPOSE: This study aimed to explore the role of long non-coding RNA (lncRNA) BDNF-AS in the progression of multiple myeloma (MM). METHODS: The expression of BDNF-AS, miR-125a-5p, and miR-125b-5p in MM serum and cell lines were detected by quantitative reverse transcriptase PCR (qRT-PCR). The binding relationships between miR-125a/b-5p and BDNF-AS or Bcl-2 were predicted by Starbase and verified by luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) staining assay. Cell migration was evaluated by wound healing assay. The expression levels of apoptosis-related proteins were evaluated by Western blot analysis. The role of BDNF-AS was also investigated in a xenograft tumor model in vivo. RESULTS: BDNF-AS was significantly upregulated, while miR-125a-5p and miR-125b-5p were downregulated in MM serum and corresponding cancer cell lines. Knockdown of BDNF-AS effectively inhibited the proliferation and migration of MM.1S and U266 cells, and co-transfection of miR-125a-5p or miR-125b-5p inhibitor and sh-BDNF-AS enhanced cell proliferation and migration compared with that in sh-BDNF-AS group. Knockdown of miR-125a-5p or miR-125b-5p significantly enhanced the proliferation and migration of MM.1S and U266 cells, and co-transfection of sh-Bcl-2 and miR-125a/b-5p inhibitor inhibited cell proliferation compared with that in miR-125a/b-5p inhibitor group. Moreover, knockdown of BDNF-AS increased the expression levels of apoptosis-related proteins (cleaved caspase 3 and cleaved PARP), while knockdown of miR-125a-5p or miR-125b-5p reduced the expression levels of these apoptosis-related proteins compared with knockdown of BDNF-AS. Furthermore, knockdown of BDNF-AS effectively suppressed MM tumor growth in vivo. CONCLUSION: Our findings revealed that knockdown of BDNF-AS inhibited the progression of MM by targeting the miR-125a/b-5p-Bcl-2 axis, indicating that BDNF-AS might serve as a novel drug target for MM.

Traditional Cantonese puerperal practices and postpartum cardiometabolism in gestational diabetes.

BACKGROUND AND OBJECTIVES: With gestational diabetes (GDM), women have a higher risk for future type 2 diabetes, and risk factors for diabetes for it are amplified. Whether this phenomenon is affected by traditional puerperal or postpartum practices among Chinese women who develop gestational diabetes is unclear. This has been explored in a Cantonese cultural setting to enable relevant risk management. METHODS AND STUDY DESIGN: Some 138 women were followed before, during and after pregnancy in accordance with Cantonese Puerperal Practices (CPP), and occurrence of GDM and exclusive breast-feeding. Body compositional and cardiometabolic information were collected. These included glucose tolerance and insulin resistance. RESULTS: During a median postpartum follow-up of 60.4 days, women with a typical CPP had a greater body weight and weight retention. With artificial feeding, women with a typical CPP had greater OGTT glycemic responses and more insulin resistance. With exclusive breast-feeding, however, no differences in postpartum cardiometabolic measurements were observed, except for a higher early-phase insulin response. CONCLUSIONS: Traditional CPP is associated with early postpartum cardiometabolic impairment in gestational diabetes, but this is avoided with breast-feeding.

Molecular cloning, characterization and expression analysis of CXCR3a and CXCR3b from Nile tilapia (Oreochromis niloticus).

The CXC chemokine receptors (CXCRs) are members of the seven transmembrane (7-TM) G-protein-coupled receptor superfamily that involves innate and adaptive immune systems. In this study, CXCR3a and CXCR3b from Nile tilapia (Oreochromis niloticus) were cloned and identified, designated as OnCXCR3a and OnCXCR3b. The open reading frames of OnCXCR3a and OnCXCR3b were 1074 and 1080 bp, encoding the predicted proteins of 357 and 359 amino acids, respectively. Multiple alignment analysis of OnCXCR3a- and OnCXCR3b-deduced protein sequences with the mammalian and bird sequences indicated the presence of typical structural features of chemokine receptors, including a 7-TM domain and conserved motifs. Quantitative real-time PCR analysis revealed that OnCXCR3a and OnCXCR3b were constitutively expressed in a wide range of tissues. When stimulated with Streptococcus agalactiae, Aeromonas hydrophila, polyinosinic:polycytidylic acid and lipopolysaccharide in vivo or in vitro on leukocytes, the mRNA levels of OnCXCR3a and OnCXCR3b were significantly upregulated. Overall, these results indicated that OnCXCR3s might be involved in host immune responses in Nile tilapia.

Antimicrobial susceptibility, multilocus sequence typing, and virulence of listeria isolated from a slaughterhouse in Jiangsu, China.

BACKGROUND: Listeria monocytogenes is one of the deadliest foodborne pathogens. The bacterium can tolerate severe environments through biofilm formation and antimicrobial resistance. This study aimed to investigate the antimicrobial susceptibility, resistance genes, virulence, and molecular epidemiology about Listeria from meat processing environments. METHODS: This study evaluated the antibiotic resistance and virulence of Listeria isolates from slaughtering and processing plants. All isolates were subjected to antimicrobial susceptibility testing using a standard microbroth dilution method. The harboring of resistant genes was identified by polymerase chain reaction. The multilocus sequence typing was used to determine the subtyping of the isolates and characterize possible routes of contamination from meat processing environments. The virulence of different STs of L. monocytogenes isolates was evaluated using a Caco-2 cell invasion assay. RESULTS: A total of 59 Listeria isolates were identified from 320 samples, including 37 L. monocytogenes isolates (62.71%). This study evaluated the virulence of L. monocytogenes and the antibiotic resistance of Listeria isolates from slaughtering and processing plants. The susceptibility of these 59 isolates against 8 antibiotics was analyzed, and the resistance levels to ceftazidime, ciprofloxacin, and lincomycin were as high as 98.31% (L. m 37; L. innocua 7; L. welshimeri 14), 96.61% (L. m 36; L. innocua 7; L. welshimeri 14), and 93.22% (L. m 35; L. innocua 7; L. welshimeri 13), respectively. More than 90% of the isolates were resistant to three to six antibiotics, indicating that Listeria isolated from meat processing environments had high antimicrobial resistance. Up to 60% of the isolates harbored the tetracycline-resistance genes tetA and tetM. The frequency of ermA, ermB, ermC, and aac(6')-Ib was 16.95, 13.56, 15.25, and 6.78%, respectively. Notably, the resistant phenotype and genotype did not match exactly, suggesting that the mechanisms of antibiotic resistance of these isolates were likely related to the processing environment. Multilocus sequence typing (MLST) revealed that 59 Listeria isolates were grouped into 10 sequence types (STs). The dominant L. monocytogenes STs were ST5, ST9, and ST121 in the slaughtering and processing plant of Jiangsu province. Moreover, ST5 subtypes exhibited high invasion in Caco-2 cells compared with ST9 and ST121 cells. CONCLUSION: The dominant L. monocytogenes ST5 persisted in the slaughtering and processing plant and had high antimicrobial resistance and invasion characteristics, illustrating a potential risk in food safety and human health.

Optical super-resonance in a customized P T-symmetric system of hybrid interaction.

We investigate the optical resonances in coupled meta-atoms with hybrid interaction pathways. One interaction pathway is the directly near-field coupling between the two meta-atoms. The other interaction pathway is via the continuum in a waveguide functioned as a common bus connecting them. We show that by properly introducing gain or loss into the meta-atoms, the hybrid optical system becomes parity-time (P T) symmetric, in which the effective coupling rate can be customized by manipulating the length of the waveguide. At the exact phase of the customized P T symmetry, the coupled meta-atoms support discrete super-resonant modes that can be observed from the transmission spectra as extremely sharp peaks. At an exception point where the eigenmodes coalesce, albeit the transmission curve is flat, a high-Q factor of the localized field in the meta-atoms can be obtained. Similarities of the super-resonance with the bound states in the continuum (BICs) are discussed. This investigation promotes our understanding about the ways in realizing high-Q optical resonance especially by manipulating the distributions of loss and gain via the concepts of P T and BICs. Many attractive applications are expected.

IgM-bearing B cell affinity subpopulations possess differential antigen sensitivity in rainbow trout.

The need for accurate assessments of in vitro generated antibody prompted examination of the effect of antigen on secreted antibody concentrations and affinities. It was found that the antigen concentrations commonly employed for in vitro stimulation were able to significantly compromise IgM titer and affinity estimates in rainbow trout. Specifically, IgM titers were underestimated with the high affinity antibodies being specifically blocked. To remedy this, pulsed antigen cultures were employed, and it was found to reveal more accurate IgM antibody titers and affinity estimates. Additionally, pulsed dose responses provided evidence that high antigen concentrations specifically suppressed high affinity B cell induction. Optimal expression of high affinity antibodies required exposure to lower concentrations of antigen. Each affinity subpopulation appeared to possess a graded sensitivity to each dose of antigen, revealing the complex dynamic for differential IgM-bearing B cell induction that is possible during a response. These results reveal not only the need for antigen removal prior to in vitro antibody secretion, but also the possible role of high zone immunological tolerance on IgM affinity maturation in rainbow trout.

Highly stable metal-organic framework UiO-66-NH2for high-performance triboelectric nanogenerators.

The high porosity, controllable size, high surface area, and chemical versatility of a metal-organic framework (MOF) enable it a good material for a triboelectric nanogenerator (TENG), and some MOFs have been incorporated in the fabrication of TENGs. However, the understanding of effects of MOFs on the energy conversion of a TENG is still lacking, which inhibits the improvement of the performance of MOF-based TENGs. Here, UiO-66-NH2MOFs were found to significantly increase the power of a TENG and the mechanism was carefully examined. The electron-withdrawing (EW) ability of Zr-based UiO-66-family MOFs was enhanced by designing the amino functionalized 1,4-terephthalic acid (1,4-BDC) as ligand. The chemically modified UiO-66-NH2was found to increase the surface roughness and surface potential of a composite film with MOFs embedded in polydimethylsiloxane (PDMS) matrix. Thus the total charges due to the contact electrification increased significantly. The composite-based TENG was found to be very durable and its output voltage and current were 4 times and 60 times higher than that of a PDMS-based TENG. This work revealed an effective strategy to design MOFs with excellent EW abilities for high-performance TENGs.

The impact of an URAT1 polymorphism on the losartan treatment of hypertension and hyperuricemia.

BACKGROUND: This study was designed to evaluate the impact of polymorphisms in the urate transporter 1 (URAT1) gene on the uricosuric action of losartan therapy in hypertensive patients suffering from hyperuricemia. METHODS: A MassARRAY approach was used to detect single nucleotide polymorphism (SNP) loci in the URAT1 and CYP2C9 genes (16 and 2 loci, respectively) in 111 patients with hypertension and hyperuricemia taking losartan and in 121 healthy controls. In addition, we compared serum urate (SUA) levels and other key clinical biochemistry indices between these two patient groups. RESULTS: We detected significant differences between the two patient groups with respect to age, SUA, urea, creatine, triglycerides, high-density lipoprotein, low-density lipoprotein, and fasting plasma glucose (all p < 0.05). In addition, we found that hypertensive patients with hyperuricemia were more likely to exhibit the rs3825016(C/T) (36.9% vs 21.5%, p = 0.03), and we determined that a 2-week treatment course with losartan was associated with significant decreases in SUA values (p < 0.001). CONCLUSION: Our findings indicate that the URAT1 rs3825016 polymorphism may influence the uricosuric action of losartan.

miR-125a suppresses malignancy of multiple myeloma by reducing the deubiquitinase USP5.

miR-125a is a microRNA that is frequently diminished in various human malignancies. However, the mechanism by which impaired miR-125a promotes cancer growth remains undefined. In this study, we investigated the role of miR-125a in the proliferation and apoptosis of multiple myeloma (MM). To do this, we used MM tissue samples (from 40 anonymous patients), normal matched control samples, and five MM-derived cell lines. We also established a mouse model of MM xenograft to explore the effect of overexpression of miR-125a on the MM growth in vivo. Quantitative real-time polymerase chain reaction revealed that the miR-125a expression was broadly reduced in MM tissues and cell lines. The impairment of miR-125a in MM tissues was functionally relevant because the overexpression of miR-125a remarkably decreased the cell viability and colony-forming activity, at least in part, by promoting apoptosis in two miR-125a-deficient MM cell lines: NCI-H929 and U266. Interestingly, we also discovered that the human gene encoding the ubiquitin-specific peptidase 5 (USP5), which is known to promote cellular deubiquitination and ubiquitin/proteasome-dependent proteolysis, was a direct transcriptional target for miR-125a to repress. More importantly, the heterologous expression of USP5 significantly reversed the growth-inhibitory effects of miR-125a on MM cells in vitro. In the mouse xenograft model, overexpressed miR-125a prominently inhibited the growth of MM tumors and concomitantly reduced the expression of USP5 in tumor tissues. These results suggest that miR-125a inhibits the expression of USP5, thereby mitigating the proliferation and survival of malignant MM cells. We propose that USP5 acts as an oncoprotein in miR-125a-missing cancers.

SHP1 tyrosine phosphatase gets involved in host defense against Streptococcus agalactiae infection and BCR signaling pathway in Nile tilapia (Oreochromis niloticus).

Src homology 2 domain-containing protein tyrosine phosphatase 1 (SHP1), a kind of protein tyrosine phosphatases (PTPs), is a critical regulator of antigen receptor signal transduction. Signal transduction of BCR is regulated by phosphatases in teleost as in mammals. In this study, SHP1 from Nile tilapia (Oreochromis niloticus) (OnSHP1) was identified and characterized, including the expression pattern against bacterial infection and regulation function in BCR signaling pathway. The open reading frame of OnSHP1 contains 1749 bp of nucleotide sequence, encoding a protein of 582 amino acids. The OnSHP1 protein was highly conversed compared to that of other species, including two amino-terminal SH2 domains at the N terminus and a PTP catalytic domain. Transcriptional expression analysis revealed that OnSHP1 was detected in all examined tissues and highly expressed in spleen. The up-regulated OnSHP1 expression was observed in peripheral blood, spleen and anterior kidney following challenge with Streptococcus agalactiae or lipopolysaccharide (LPS) in vivo, as well as that displayed in leukocytes stimulated with S. agalactiae or LPS in vitro. Further, after induction with mouse anti-tilapia IgM monoclonal antibody in vitro, OnSHP1 was significantly up-regulated in leukocytes. When spleen leukocytes treated with PTP Inhibitor II in vitro, the phosphorylation level of OnSHP1 at the phosphorylation sites (Y535 and Y557) and the cytoplasmic free Ca2+ concentration were up-regulated significantly. Overall, the findings of this study indicate that SHP1 gets involved in host defense against bacterial infection and BCR signaling pathway in Nile tilapia.

Molecular characterization and expression analysis of chemokine (CXCL12) from Nile tilapia (Oreochromis niloticus).

Chemokines are a class of small molecular weight cytokines of 6-14 kDa, exerting important roles in the regulation of various inflammatory diseases and immune cell migration. In this study, we have identified the CXCL12 gene from Nile tilapia (Oreochromis niloticus), including CXCL12a (OnCXCL12a) and CXCL12b (OnCXCL12b). The open reading frames of OnCXCL12a and OnCXCL12b are 309 and 297 bp, encoding 102 and 98 amino acids, respectively. Multiple alignment showed that OnCXCL12a and OnCXCL12b have characteristics of CXC chemokines and share high identity with CXCL12 amino acid sequences from the known species. Tissue distribution in the healthy fish indicated that OnCXCL12a and OnCXCL12b expressed in all examined tissues, with the highest expression in muscle and anterior kidney, respectively. After challenged by Streptococcus agalactiae, Poly(I:C) and LPS in vivo and in vitro, OnCXCL12 is transcriptionally up-regulated in immune tissues and cells significantly. The recombinant OnCXCL12 proteins, (r)OnCXCL12a and (r)OnCXCL12b, enhance the release of nitric oxide and increase the expression of inflammatory cytokines (TNF-α, IL-6, and IL-10) in anterior kidney leukocytes, as well as exhibit chemotactic activity for leukocytes from anterior kidney. Summarizing, these results indicate that OnCXCL12 is involved in the immune response of Nile tilapia against pathogen infection and may play an important role in mediating inflammatory response.

A Siglec-1-like lectin from Nile tilapia (Oreochromis niloticus) possesses functions of agglutination and mediation of macrophage phagocytic activity.

Siglec-1, one of the sialic acid-binding immunoglobulin-type lectins, is closely related to the recognition of host-pathogen and cell-cell interactions in the adaptive and innate immune systems. In this communication, a Siglec-1-like gene (OnSiglec-1-like) from Nile tilapia (Oreochromis niloticus) was analyzed. Relative expression revealed that the OnSiglec-1-like was expressed in all tested tissues, and the highest expression was found in the anterior kidney. Upon Streptococcus agalactiae (S. agalactiae) infection, the expression of OnSiglec-1-like was up-regulated in anterior kidney and spleen significantly in vivo. Additionally, the same phenomenon was observed in anterior kidney leukocytes upon LPS and S. agalactiae challenges as well in vitro. Western-blotting and ELISA analyses revealed that recombinant OnSiglec-1-like protein possessed high binding activity to LTA, LPS and S. agalactiae. Further, the recombinant OnSiglec-1-like was able to agglutinate S. agalactiae. Moreover, with the digestion of specific sialidase, the phagocytic ability of macrophages to S. agalactiae was greatly enhanced. Taken together, these results indicated that the Siglec-1-like possesses conserved functions of agglutination and promotion of macrophage phagocytic activity in Nile tilapia.