•Anti-PcrV titers of non-cystic fibrosis patient with/without Pseudomonas aeruginosa respiratory tract infections were investigated.•Anti-PcrV titers were significantly higher in the P. ...aeruginosa-infected patients.•Those patients with a lower anti-PcrV titer in P. aeruginosa infection occasionally present a poorer outcome.•Patients with low anti-PcrV titers and refractory P. aeruginosa infections need to be monitored closely.
The epidemiology and role of the anti-PcrV titer in non-cystic fibrosis patients with Pseudomonas aeruginosa airway tract infections is not fully understood. This study was performed to compare the anti-PcrV titers of patients with and without P. aeruginosa respiratory tract infections.
This prospective cohort study was conducted at Hokkaido University Hospital in Japan. Participants had blood and sputum specimens collected on admission. They were divided into two groups based on their sputum culture results. Those with a P. aeruginosa infection were assigned to the P. aeruginosa (PA) group and those without a P. aeruginosa infection were assigned to the non-PA group. Serum anti-PcrV titers were measured using a validated ELISA.
Of the 44 participants, 15 were assigned to the PA group and 29 were assigned to the non-PA group. In the PA group, 10/15 participants (66.7%) had an anti-PcrV titer >1000ng/ml compared to 3/29 participants (10.3%) in the non-PA group (p<0.001). In the PA group, two of the five participants with an anti-PcrV titer <1000 ng/ml died of recurrent P. aeruginosa pneumonia; the other three participants did not develop pneumonia.
The anti-PcrV titers in participants with P. aeruginosa infection varied considerably. Patients with low anti-PcrV titers and refractory P. aeruginosa infections need to be monitored closely.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Medium- and long-chain free fatty acids (FFAs) are energy source for whole body and biological metabolites and components. In these decades, some research groups have reported that the biological ...functions of medium- to long-chain FFAs are exerted through G-protein coupled receptor designated free fatty acid receptor (FFAR). As the medium- to long-chain FFAs-activated FFARs, FFA1 and FFA4 are reported to be expressed widely in whole body and regulate various physiological processes. FFA1 expressed in pancreatic β-cells has been shown to be involved in insulin secretion. FFA4 expressed in intestine, adipocytes, and macrophages has been shown to be involved in incretin secretion, differentiation, and anti-inflammatory effect, respectively. These physiological functions have been focused on the treatment of metabolic disorders. In addition, these receptors have been also reported to be expressed in several other tissues such as intestine for FFA1, and tongue and stomach for FFA4. The recent functional studies indicated that they also contributed to energy homeostasis. Further, the number of synthetic compounds of FFA1 and FFA4 strongly promoted the physiological characterization of the receptors and their own therapeutic utility. In this article, we will discuss the recent progress regarding the therapeutic potential of these receptors and its ligands.
Aim
Tumor necrosis factor‐α (TNF‐α) is a pleiotropic cytokine involved in various inflammatory diseases. The only production of TNF‐α in the liver is thought to be from hepatic macrophages known as ...Kupffer cells, predominantly in response to bacterial lipopolysaccharide (LPS).
Methods
Primary cultured rat hepatocytes were used to analyze TNF‐α expression in response to the pro‐inflammatory cytokine, interleukin‐1β (IL‐1β). Livers of rats subjected to LPS‐induced endotoxemia were analyzed.
Results
Immunocytochemistry and enzyme‐linked immunosorbent assays demonstrated that IL‐1β‐treated rat hepatocytes secreted TNF‐α, and RNA analyses indicated that TNF‐α mRNA was induced specifically by IL‐1β. Northern blot analysis showed that not only mRNA, but also a natural antisense transcript (asRNA), was transcribed from the rat Tnf gene in IL‐1β‐treated hepatocytes. TNF‐α was detected in the hepatocytes of LPS‐treated rats. Both TNF‐α mRNA and asRNA were expressed in the hepatocytes of LPS‐treated rats, human hepatocellular carcinoma and human monocyte/macrophage cells. To disrupt the interaction between TNF‐α asRNA and TNF‐α mRNA, sense oligonucleotides corresponding to TNF‐α mRNA were introduced into rat hepatocytes resulting in significantly increased levels of TNF‐α mRNA. One of these sense oligonucleotides increased a half‐life of TNF‐α mRNA, suggesting that the TNF‐α asRNA may reduce the stability of TNF‐α mRNA.
Conclusion
IL‐1β‐stimulated rat hepatocytes are a newly identified source of TNF‐α in the liver. TNF‐α mRNA and asRNA are expressed in rats and humans, and the TNF‐α asRNA reduces the stability of the TNF‐α mRNA. Hepatocytes and TNF‐α asRNA may be therapeutic targets to regulate levels of TNF‐α mRNA.
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BFBNIB, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UL, UM, UPUK
The prevalence of extended-spectrum β-lactamase (ESBL) in Enterobacteriaceae has been increasing worldwide. The aims of this study were to determine the prevalence of ESBLs among clinical isolates of ...Escherichia coli obtained from 2000 to 2010 in Japan, and to characterize the sequence type (ST) and antimicrobial susceptibility of the bla(CTX-M)-carrying strains. The genes for β-lactamases were determined by conventional PCR and sequencing, and the antimicrobial susceptibility test was performed by the broth microdilution method. Among the 948 strains, 35 were judged as ESBL-positive strains. The positive rates ranged from 0.6% to 3.9% until 2008, but surged to 10.3% in 2010. Thirty-three of them carried bla(CTX-M), but all were negative for ESBL-type bla(TEM) and bla(SHV). bla(CTX-M-14) was the most prevalent (18/33) among bla(CTX-M)-carrying strains, followed by bla(CTX-M-15) (7/33) of which five were isolated in 2008 and 2010. Additionally, bla(CTX-M-27) appeared in 2010 for the first time in this study and accounted for more than a third of the bla(CTX-M)-carrying strains. From the MLST analysis, ST131 known as a world pandemic clone, has been predominantly isolated since 2006. The major types of ESBLs carried by ST131 strains clearly shifted from bla(CTX-M-14) to bla(CTX-M-15) and/or bla(CTX-M-27) between 2006 and 2010. Most of these isolates were still susceptible to doripenem, latamoxef (moxalactam), flomoxef and cefmetazole. Our results suggest that a change of the dominant type of ESBL among Enterobacteriaceae is currently in progress in Japan, and therefore further periodic surveillance is needed.
Abstract To compare the risk of acquiring in vitro resistance between doripenem and tazobactam/piperacillin by CTX-M-15-producing Escherichia coli , the in vitro frequency of resistance was ...determined. Four strains carrying multiple β-lactamases such as blaOXA-1 or blaCTX-M-27 as well as blaCTX-M-15 and blaTEM-1 were used. No resistant colonies appeared on doripenem-containing plates, whereas resistant colonies were obtained from three of four test strains against tazobactam/piperacillin using agar plate containing 8- to 16-fold MIC of each drug. These three acquired tazobactam/piperacillin-resistant strains were not cross-resistant to doripenem, and they showed 1.9- to 3.1-fold higher piperacillin-hydrolysis activity compared to those of each parent strain. The change of each β-lactamase mRNA expression measured by real-time PCR varied among three resistant strains. One of three tazobactam/piperacillin-resistant strains with less susceptibility to ceftazidime overexpressed both blaCTX-M-15 and blaTEM-1 , and the other two strains showed higher mRNA expression of either blaTEM-1 or blaOXA-1 . These results demonstrate that multiple β-lactamases carried by CTX-M-15-producing E. coli contributed to the resistance to tazobactam/piperacillin. On the other hand, these resistant strains maintained susceptibility to doripenem. The risk of acquiring in vitro resistance to doripenem by CTX-M-15-producing E. coli seems to be lower than that to tazobactam/piperacillin.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
GPR40 is a G protein-coupled receptor (GPCR) whose endogenous ligands have recently been identified as medium- and long-chain
free fatty acids (FFAs), and it is thought to play an important role in ...insulin release. Despite recent research efforts,
much still remains unclear in our understanding of its pharmacology, mainly because the receptor-ligand interaction has not
been analyzed directly. To study the pharmacology of GPR40 in a more direct fashion, we developed a flow cytometry-based binding
assay. FLAG-tagged GPR40 protein was expressed in Sf9 cells, solubilized, immobilized on immunomagnetic beads, and labeled
with the fluorescent probe C1-BODIPY-C12. Flow cytometry analysis showed that C1-BODIPY-C12 specifically labels a single class
of binding site in a saturable and reversible manner with an apparent dissociation constant of â¼3 μM. The FFAs that activate
GPR40 competed with C1-BODIPY-C12 binding; thus, medium- to long-chain FFAs could compete, whereas short-chain FFAs and methyl
linoleate had no inhibitory effect. Furthermore, ligands that are known to activate GPR40 competed for binding in a concentration-dependent
manner. All the ligands that inhibited the binding promoted phosphorylation of extracellular signal-regulated kinase (ERK)-1/2
in human embryonic kidney (HEK) 293 cells that expressed GPR40 and Ca 2+ i responses in mouse insulinoma (MIN6) cells that natively express GPR40; however, pioglitazone, a thiazolidinedione that failed
to compete for the binding, did not activate ERK or Ca 2+ i response. This study showed that a flow cytometry-based binding assay can successfully identify direct interactions between
GPR40 and its ligands. This approach would be of value in studying the pharmacology of GPCRs.
► Human iNOS antisense transcripts are expressed in the liver and colon cancers. ► Human iNOS antisense transcript corresponds to 3′ untranslated region of iNOS mRNA. ► Sense oligonucleotides ...corresponding to iNOS mRNA degrade iNOS mRNA. ► Modified iNOS sense oligonucleotides greatly reduce iNOS mRNA levels. ► iNOS sense oligonucleotides may be used to treat human diseases including cancers.
Natural antisense transcripts (asRNAs) are frequently transcribed from mammalian genes. Recently, we found that non-coding asRNAs are transcribed from the 3′ untranslated region (3′UTR) of the rat and mouse genes encoding inducible nitric oxide synthase (iNOS), which catalyzes the production of the inflammatory mediator nitric oxide. The iNOS asRNA stabilizes iNOS mRNA by interacting with the mRNA 3′UTR. Furthermore, single-stranded ‘sense’ oligonucleotides corresponding to the iNOS mRNA sequence were found to reduce iNOS mRNA levels by interfering with mRNA–asRNA interactions in rat hepatocytes. This method was named natural antisense transcript-targeted regulation (NATRE) technology. In this study, we detected human iNOS asRNA expressed in hepatocarcinoma and colon carcinoma tissues. The human iNOS asRNA harbored a sequence complementary to an evolutionarily conserved region of the iNOS mRNA 3′UTR. When introduced into hepatocytes, iNOS sense oligonucleotides that were modified by substitution with partial phosphorothioate bonds and locked nucleic acids or 2′-O-methyl nucleic acids greatly reduced levels of iNOS mRNA and iNOS protein. Moreover, sense oligonucleotides and short interfering RNAs decreased iNOS mRNA to comparable levels. These results suggest that NATRE technology using iNOS sense oligonucleotides could potentially be used to treat human inflammatory diseases and cancers by reducing iNOS mRNA levels.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
In this article, we evaluate the noise and vibration in a permanent-magnet synchronous motor (PMSM) with distributed winding driven by voltage source pulsewidth modulation (PWM) inverters. Although ...the PMSM is a widely used device in a variety of fields, it tends to generate electromagnetic noise and vibration due to the PWM inverters. We describe the frequency of the electromagnetic vibration to estimate the worst point of the carrier electromagnetic vibration of dc voltage fluctuation. The PWM voltage harmonics and the modulation degree of dc voltage fluctuation were investigated through a theoretical analysis of the PWM voltage harmonics. It was found that a modulation degree near 0.56 was the maximum point of the doubled component of the carrier frequency <inline-formula><tex-math notation="LaTeX">2f_{c}</tex-math></inline-formula>. Compared with the dc voltage fluctuation at this point, the dominant component of the PWM voltage harmonics was near frequency <inline-formula><tex-math notation="LaTeX">f_{c}</tex-math></inline-formula> at a lower voltage and near frequency <inline-formula><tex-math notation="LaTeX">2f_{c}</tex-math></inline-formula> at a higher voltage. These tendencies of the carrier electromagnetic vibration of dc voltage fluctuation are also verified in an experiment.
GPR40 is G protein-coupled receptor whose endogenous ligands have recently been identified as free fatty acids (FFAs), and it has been implicated to play an important role in FFA-mediated enhancement ...of glucose-stimulated insulin release. We have developed a monoclonal antibody against the extracellular domain of GPR40. Specificity of the antibody was demonstrated by immunoprecipitation and cell surface staining using GPR40-transfected cells. GPR40 immunoreactivity was highly abundant in mouse pancreatic β-cells and splenocytes, THP-1 cells, and human peripheral blood mononuclear cells. The anti-GPR40 monoclonal antibody should prove valuable for further studying the function of this nutrient sensing receptor.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
This paper describes the spatial mode, frequencies, the voltage reference update timing, and their interaction of pulsewidth modulation (PWM) harmonics, focusing on a distributed winding permanent ...magnet synchronous motor. We investigated the electromagnetic force generating the noise by using a couple of analyses between a circuit simulator and a two-dimensional finite element analysis. It was found that the electromagnetic noise and vibration result from a radial electromagnetic force spaced at the spatial zeroth mode of <inline-formula><tex-math notation="LaTeX">f_{c}\pm 3f_{1},f_{c}</tex-math></inline-formula>, and <inline-formula><tex-math notation="LaTeX">{{2}}f_{c}</tex-math> </inline-formula> (<inline-formula><tex-math notation="LaTeX">f_{c}</tex-math></inline-formula>: carrier frequency, <inline-formula><tex-math notation="LaTeX">f_{1}</tex-math></inline-formula>: fundamental frequency of the current). We have shown that the radial electromagnetic force of frequency <inline-formula><tex-math notation="LaTeX">f_{c}\pm 3f_{1}</tex-math></inline-formula> and <inline-formula><tex-math notation="LaTeX">{{2}}f_{c}</tex-math> </inline-formula> results from PWM harmonics. Also, the radial electromagnetic force of frequency <inline-formula> <tex-math notation="LaTeX">f_{c}</tex-math></inline-formula> evidently results from the voltage reference update timing of <inline-formula><tex-math notation="LaTeX">{{1/ }}f_{c}</tex-math></inline-formula>. The electromagnetic noise and the spatial mode are demonstrated in the experiment.
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