HutZ is required for biofilm formation and contributes to the pathogenicity of Edwardsiella piscicida

International audienceAbstractEdwardsiella piscicida is a severe fish pathogen. Haem utilization systems play an important role in bacterial adversity adaptation and pathogenicity. In this study, a speculative haem utilization protein, HutZEp, was characterized in E. piscicida. hutZEp is encoded with two other genes, hutW and hutX, in an operon that is similar to the haem utilization operon hutWXZ identified in V. cholerae. However, protein activity analysis showed that HutZEp is probably not related to hemin utilization. To explore the biological role of HutZEp, a markerless hutZEp in-frame mutant strain, TX01ΔhutZ, was constructed. Deletion of hutZEp did not significantly affect bacterial growth in normal medium, in iron-deficient conditions, or in the presence of haem but significantly retarded bacterial biofilm growth. The expression of known genes related to biofilm growth was not affected by hutZEp deletion, which indicated that HutZEp was probably a novel factor promoting biofilm formation in E. piscicida. Compared to the wild-type TX01, TX01ΔhutZ exhibited markedly compromised tolerance to acid stress and host serum stress. Pathogenicity analysis showed that inactivation of hutZEp significantly impaired the ability of E. piscicida to invade and reproduce in host cells and to infect host tissue. In contrast to TX01, TX01ΔhutZ was defective in blocking host macrophage activation. The expression of hutZEp was directly regulated by the ferric uptake regulator Fur. This study is the first functional characterization of HutZ in a fish pathogen, and these findings suggested that HutZEp is essential for E. piscicida biofilm formation and contributes to host infection

Dual roles of neutrophils in metastatic colonization are governed by the host NK cell status.

The role of neutrophils in solid tumor metastasis remains largely controversial. In preclinical models of solid tumors, both pro-metastatic and anti-metastatic effects of neutrophils have been reported. In this study, using mouse models of breast cancer, we demonstrate that the metastasis-modulating effects of neutrophils are dictated by the status of host natural killer (NK) cells. In NK cell-deficient mice, granulocyte colony-stimulating factor-expanded neutrophils show an inhibitory effect on the metastatic colonization of breast tumor cells in the lung. In contrast, in NK cell-competent mice, neutrophils facilitate metastatic colonization in the same tumor models. In an ex vivo neutrophil-NK cell-tumor cell tri-cell co-culture system, neutrophils are shown to potentially suppress the tumoricidal activity of NK cells, while neutrophils themselves are tumoricidal. Intriguingly, these two modulatory effects by neutrophils are both mediated by reactive oxygen species. Collectively, the absence or presence of NK cells, governs the net tumor-modulatory effects of neutrophils

Use of serial analysis of gene expression to reveal the specific regulation of gene expression profile in asthmatic rats treated by acupuncture

<p>Abstract</p> <p>Background</p> <p>Asthma has become an important public health issue and approximately 300 million people have suffered from the disease worldwide. Nowadays, the use of acupuncture in asthma is increasing. This study intended to systematically analyze and compare the gene expression profiles between the asthmatic and acupuncture-treated asthmatic rat lung, and tried to gain insight into the molecular mechanism underlying the early airway response (EAR) phase of asthma treated by acupuncture.</p> <p>Methods</p> <p>Four tag libraries of serial analysis of gene expression (SAGE) were established from lung tissues of control rats (CK), asthmatic rats (AS), asthmatic rats treated by acupuncture (ASAC), and control rats treated by acupuncture (CKAC). Bioinformatic analyses were carried out by using the methods including unsupervised hierarchical clustering, functional annotation tool of the database for annotation, visualization, and integrated discovery (DAVID), gene ontology (GO) tree machine, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis.</p> <p>Results</p> <p>There were totally 186 differentially expressed tags (P < 0.05, P_CK/AS) between the libraries of CK and AS, 130 differentially expressed tags between libraries of AS/ASAC (P < 0.05, P_AS/ASAC), and 144 differentially expressed tags between libraries of CK/CKAC (P < 0.05, P_CK/CKAC). The gene expression profiles of AS and ASAC were more similar than other libraries via unsupervised SAGE clustering. By comparison of P_CK/ASand P_AS/ASAC, the DAVID genes functional classification was found to be changed from "immune response" to "response to steroid hormone stimulus", and the GO term "antigen processing and presentation of peptide antigen" disappeared in P_AS/ASAC. Totally 3 same KEGG pathways were found among the three groups. Moreover, 21 specific tags of the acupuncture in treating asthma were detected using Venn diagrams.</p> <p>Conclusion</p> <p>Our SAGE research indicates that the gene expression profile of the EAR phase of asthma could be effectively and specifically regulated by acupuncture, which suggests that the gene expression of immune response and steroid hormone may play an important role in the treatment.</p

Plasma Lipoprotein-associated Phospholipase A2 in Patients with Metabolic Syndrome and Carotid Atherosclerosis

<p>Abstract</p> <p>Background</p> <p>Lipoprotein-associated phospholipase A₂(Lp-PLA₂) is a recently identified and potentially useful plasma biomarker for cardiovascular and atherosclerotic diseases. However, the correlation between the Lp-PLA₂activity and carotid atherosclerosis remains poorly investigated in patients with metabolic syndrome (MetS). The present study aimed to evaluate the potential role of Lp-PLA₂as a comprehensive marker of metabolic syndrome in individuals with and without carotid atherosclerosis.</p> <p>Methods</p> <p>We documented 118 consecutive patients with MetS and 70 age- and sex-matched healthy subjects served as controls. The patients were further divided into two groups: 39 with carotid plaques and 79 without carotid plaques to elucidate the influence of Lp-PLA₂on carotid atherosclerosis. The plasma Lp-PLA₂activity was measured by using ELISA method and carotid intimal-media thickness (IMT) was performed by ultrasound in all participants.</p> <p>Results</p> <p>Lp-PLA₂activity was significantly increased in MetS subgroups when compared with controls, and was higher in patients with carotid plaques than those without plaques (<it>P </it>< 0.05). Furthermore, we found that significant difference in Lp-PLA₂was obtained between patients with three and four disorders of metabolic syndrome (<it>P </it>< 0.01). Age (β = 0.183, <it>P </it>= 0.029), LDL-cholesterol (β = 0.401, <it>P </it>= 0.000) and waist-hip ratio (β = 0.410, <it>P </it>= 0.000) emerged as significant and independent determinants of Lp-PLA₂activity. Multiple stepwise regression analysis revealed that LDL-cholesterol (β = 0.309, <it>P </it>= 0.000), systolic blood pressure (β = 0.322, <it>P </it>= 0.002) and age (β = 0.235, <it>P </it>= 0.007) significantly correlated with max IMT, and Lp-PLA₂was not an independent predictor for carotid IMT.</p> <p>Conclusions</p> <p>Lp-PLA₂may be a modulating factor for carotid IMT via age and LDL-cholesterol, not independent predictor in the pathophysiological process of carotid atherosclerosis in patients with MetS.</p

Loop-Mediated Isothermal Amplification Assay Targeting the MOMP Gene for Rapid Detection of Chlamydia psittaci Abortus Strain

For rapid detection of the Chlamydia psittaci abortus strain, a loop-mediated isothermal amplification (LAMP) assay was developed and evaluated in this study. The primers for the LAMP assay were designed on the basis of the main outer membrane protein (MOMP) gene sequence of C. psittaci. Analysis showed that the assay could detect the abortus strain of C. psittaci with adequate specificity. The sensitivity of the test was the same as that of the nested-conventional PCR and higher than that of chick embryo isolation. Testing of 153 samples indicated that the LAMP assay could detect the genome of the C. psittaci abortus strain effectively in clinical samples. This assay is a useful tool for rapid diagnosis of C. psittaci infection in sheep, swine and cattle

Cloning and expression profiling of the VLDLR gene associated with egg performance in duck (Anas platyrhynchos)

<p>Abstract</p> <p>Background</p> <p>The very low density lipoprotein receptor gene (<it>VLDLR</it>), a member of the low density lipoprotein receptor (<it>LDLR</it>) gene family, plays a crucial role in the synthesis of yolk protein precursors in oviparous species. Differential splicing of this gene has been reported in human, rabbit and rat. In chicken, studies showed that the VLDLR protein on the oocyte surface mediates the uptake of yolk protein precursors into growing oocytes. However, information on the <it>VLDLR </it>gene in duck is still scarce.</p> <p>Methods</p> <p>Full-length duck <it>VLDLR </it>cDNA was obtained by comparative cloning and rapid amplification of cDNA ends (RACE). Tissue expression patterns were analysed by semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR). Association between the different genotypes and egg performance traits was investigated with the general linear model (GLM) procedure of the SAS^®software package.</p> <p>Results</p> <p>In duck, two <it>VLDLR </it>transcripts were identified, one transcript (variant-a) containing an O-linked sugar domain and the other (variant-b) not containing this sugar domain. These transcripts share ~70 to 90% identity with their counterparts in other species. A phylogenetic tree based on amino acid sequences showed that duck VLDLR proteins were closely related with those of chicken and zebra finch. The two duck <it>VLDLR </it>transcripts are differentially expressed i.e. <it>VLDLR-a </it>is mainly expressed in muscle tissue and <it>VLDLR-b </it>in reproductive organs. We have localized the duck <it>VLDLR </it>gene on chromosome Z. An association analysis using two completely linked SNP sites (T/C at position 2025 bp of the ORF and G/A in intron 13) and records from two generations demonstrated that the duck <it>VLDLR </it>gene was significantly associated with egg production (P < 0.01), age of first egg (P < 0.01) and body weight of first egg (P < 0.05).</p> <p>Conclusions</p> <p>Duck and chicken <it>VLDLR </it>genes probably perform similar function in the development of growing oocytes and deposition of yolk lipoprotein. Therefore, <it>VLDLR </it>could be a candidate gene for duck egg performance and be used as a genetic marker to improve egg performance in ducks.</p