Escherichia coli (E. coli) is a frequent gram-negative bacterium that causes nosocomial infections, affecting more than 100 million patients annually worldwide. Bacterial lipopolysaccharide (LPS) from E. coli binds to toll-like receptor 4 (TLR4) and its co-receptor’s cluster of differentiation protein 14 (CD14) and myeloid differentiation factor 2 (MD2), collectively known as the LPS receptor complex. LPCAT2 participates in lipid-raft assembly by phospholipid remodelling. Previous research has proven that LPCAT2 co-localises in lipid rafts with TLR4 and regulates macrophage inflammatory response. However, no published evidence exists of the influence of LPCAT2 on the gene expression of the LPS receptor complex induced by smooth or rough bacterial serotypes. We used RAW264.7—a commonly used experimental murine macrophage model—to study the effects of LPCAT2 on the LPS receptor complex by transiently silencing the LPCAT2 gene, infecting the macrophages with either smooth or rough LPS, and quantifying gene expression. LPCAT2 only significantly affected the gene expression of the LPS receptor complex in macrophages infected with smooth LPS. This study provides novel evidence that the influence of LPCAT2 on macrophage inflammatory response to bacterial infection depends on the LPS serotype, and it supports previous evidence that LPCAT2 regulates inflammatory response by modulating protein translocation to lipid rafts.
Despite extensive investigations, an effective treatment for sepsis remains elusive and a better understanding of the inflammatory response to infection is required to identify potential new targets for therapy. In this study we have used RNAi technology to show, for the first time, that the inducible lysophosphatidylcholine acyltransferase 2 (LPCAT2) plays a key role in macrophage inflammatory gene expression in response to stimulation with bacterial ligands. Using siRNA- or shRNA-mediated knockdown, we demonstrate that, in contrast to the constitutive LPCAT1, LPCAT2 is required for macrophage cytokine gene expression and release in response to TLR4 and TLR2 ligand stimulation but not for TLR-independent stimuli. In addition, cells transfe
... Show Morehe present work, among other previous studies done in our lab, aimed to highlight the histopathological effect of S. xylosus peptidoglycan in comparison to LPS of E. coli. Materials and methods: One hundred and fifty urine specimens were collected from urinary tract infection patients visiting Baghdad hospitals. The histopathological effects of S. xylosus S24 peptidoglycan was studied in the urinary tract of female mice by injecting 5 animal groups at the following concentrations: 1000, 2000, 3000, 4000, and 5000 µg/mL. Another 5 groups were injected with 10, 25, 50, 75, and 100 ng/mL of E. coli (serotype 0128:B12) LPS. Results: Ten isolates were confirmed to be Staphylococcus xylosus. Histopathological study showed different pathological
... Show MoreLeishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no differen
... Show MoreOne hundred twelve urine samples were collected from Baghdad hospitals and examined by different identification techniques. Seventy isolates (62.5%) were diagnosed as Escherichia coli after microscopic and cultural identifications. The result of PCR product electrophoresis on the isolates showed that thirteen isolates (18.57%) have Pap E gene which are uropathogenic E. coli. Antibiotic susceptibility test was done, and four high resistant strains were mixed with aqueous extract of Quercus infectoria plant in 96 well ELISA plate and incubated for different times. After 0, 6, and 12 hr. of incubation, the effect of the plant extract on the bacterial growth was determined by ELISA reader, and the effect on the expression of P
... Show MoreAn enzyme linked immunosorbent assay (ELISA) for the detection and quantitation of human immunoglobulin G (IgG) antibodies against vero- cytotoxine (VT) producing Escherichia coli serogroup O157:H7 was produced. E. coli O157: H7 lipopolysaccharide was extracted from locally isolated strains by using hot phenol- water method, followed by partial purification using gel filtration chromatography by sepharose- 4B. The purity of the lipopolysaccharide was checked by measuring the protein and nucleic acid content and then used as antigen. Four isolates of vero- cytotoxin producing E. coli serogroup O157:H7 was obtained by culturing 350 stool samples from children suffering from bloody diarrhea. These isolates were identified on bacteriological, s
... Show MoreInflammatory control is essential to diminish injury and make renal injury treatment simpler. Proposed therapeutics have primarily targeted pro-inflammatory variables. Juniperus oxycedrus was frequently used to treat a variety of infectious disorders, hyperglycemia, obesity, TB, bronchitis, inflammation, and pneumonia. Juniperus oxycedrus twigs and leaves were defatted with n-hexane using Soxhlet apparatus then the residue of plant material dried and re-extracted sequentially by two different solvents Ethylacetate and methanol. The pro-inflammatory markers IL-1 and iNOS, as well as the potential kidney biomarker KIM-1, TNF-α, and transcription factor NF-KB were measured using the RealTime Quantitative qPCR method. The results showed that J
... Show MoreAim: To evaluation the effect of Lactobacillus acidophilus on Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 with detection of some virulence factors. Methods: Two hundred and fifty specimens (stool) from children under five years for both sexes were collected from some hospitals. All isolates were diagnosed according to morphological characteristics, biochemical tests. Monoplex pattern of PCR was used also for detection different genes in (7) Escherichia coli )O157:H7 (isolates; include 16SrRNA, eae, lifA, Stx1,Stx2 that encoded for ribosomal RNA, intimin, lymphocyte inhibitory factor, shiga toxins. Three types of probiotics strains were obtained, Lactobacillus fermentum, Lactobacillus plantarum and Lactobacillus acidophilus (A
... Show MoreIn the resent years, there is a robust scientific interest in discovery of new anti-septic and anti-oxidant naturally products with no/or limited side effects. The current study aimed to investigate the protective role of the quercetin on inflammations induced by lipopolysaccharide (LPS) in male mice A number of criteria included i.e. liver and spleen index and IL-6 and IL1-β cytokines level in spleen homogenate were considered. Sixty male mice (8-9 week age) was divided into six groups and treated for 5 days as the following: the first group represented control, the second and third group were injected with 5, 10 mg/kg b.w doses of quercetin respectively. While the fourth and fifth groups were co-treatment with (5, 10 mg/kg b.w.) intraper
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