Diagnostic value of apoptosis biomarkers in severe sepsis-A pilot study.

Severe sepsis is associated with significant mortality and massive immune cell lose, or apoptosis. It is unclear whether plasma apoptosis biomarkers could be used as a diagnostic test for severe sepsis. Forty patients with severe sepsis and 35 healthy controls were enrolled. The percentage and apoptosis of monocytes and lymphocytes were detected by flow cytometric analysis. Plasma levels of tumor necrosis factor (TNF)-α, soluble TNF receptor (sTNFR), soluble Fas (sFas), Fas ligand (FasL), caspase-1, and procalcitonin (PCT) were measured. Plasma caspase-1 level was positively correlated with CD4 lymphocyte apoptosis in controls and patients, and with CD8 lymphocyte apoptosis in all subjects. Plasma FasL level was negatively correlated with CD4 and CD8 lymphocyte apoptosis in all subjects. The sFas/FasL ratio was positively correlated with CD4 and CD8 lymphocyte apoptosis and negatively with monocyte apoptosis in all subjects. Compared with PCT, caspase-1, FasL, and sFas/FasL ratio had better negative predictive value and likelihood ratio for a negative test. PCT had better positive predictive value and likelihood ratio for a positive test. This work demonstrated caspase-1, FasL, and sFas/FasL ratio could be candidates for diagnosis of severe sepsis and their diagnostic value was not inferior to that of PCT.

Effects of Elderberry Juice from Different Genotypes on Oxidative and Inflammatory Responses in Microglial Cells.

Many species of berries are nutritious food and offer health benefits. However, among the different types of berries, information on health effects of American elderberries (Sambucus nigra subsp. canadensis) has been lacking and little is known about whether elderberry consumption can confer neuroprotective effects on the central nervous system. Microglial cells constitute a unique class of immune cells and exhibit characteristic properties to carry out multifunctional duties in the brain. Activation of microglial cells has been implicated in brain injury and in many types of neurodegenerative diseases. Our recent studies demonstrated the ability for endotoxin (lipopolysaccharide, LPS) and interferon gamma (IFNγ) to induce reactive oxygen species (ROS) and nitric oxide (NO) in murine microglial cells (BV-2) through activating NADPH oxidase and the MAPK pathways. In this study, BV-2 microglial cells were used to examine effects of elderberry juice obtained from different genotypes on oxidative and inflammatory responses induced by LPS and IFNγ. Results show that 'Wyldewood' extract demonstrated antioxidant properties by inhibiting IFNγ-induced ROS production and p-ERK1/2 expression. On the other hand, most juice extracts exerted small effects on LPS-induced NO production and some extracts showed an increase in NO production upon stimulation with IFNγ. The disparity of responses on ROS and NO production from different extracts suggests possible presence of unknown endogenous factor(s) in the extract in promoting the IFNγ-induced iNOS synthesis pathway.

Expression of short-form oncostatin M receptor as a decoy receptor in lung adenocarcinomas.

Oncostatin M (OSM) is a member of the interleukin-6 (IL-6) family of cytokines, and binds to the OSM receptor (OSMR) to inhibit cancer growth. Four forms of OSMR have been identified: leukemia inhibitory factor receptor (LIFR), OSMR beta, short-form OSMR (OSMRs) and soluble OSMR (sOSMR). In this study, we examined the type and expression of OSMR in lung adenocarcinomas (LADCs). Expression of OSMR was determined by reverse transcription-polymerase chain reaction (RT-PCR), immunoblotting, immunohistochemistry and confocal immunofluorescent microscopy (CIM). Our results showed that, among the four forms of OSMR, OSMRs was mainly expressed in LADC, and expression level of OSMRs correlated with patient survival. CIM revealed that OSMRs was localized on the cell membrane of LADC cell lines in vitro. OSMRs acts as a decoy receptor by reducing the inhibitory effect of OSM on cell growth. Decrease in OSMRs expression by siRNA increased cell sensitivity to OSM, and ectopic expression of OSMRs reduced cell sensitivity to OSM. These results suggest that expression of OSMRs, which operates as a decoy receptor for OSM, is correlated with disease progression and adverse prognosis in patients with LADC.

Comparison of interferon-gamma response between tuberculosis and non-tubercular pneumonia.

OBJECTIVE AND DESIGN: Macrophages aided by interferon-gamma (IFN-gamma) are vital to controlling Mycobacterium tuberculosis (M. tuberculosis) infection. Although numerous studies have compared IFN-gamma response between tubercular patients and healthy controls, no studies have investigated IFN-gamma response in patients with pulmonary tuberculosis and non-tubercular pneumonia. The aim of this work was to examine the difference in IFN-gamma response between patients with tuberculosis and non-tubercular pneumonia. METHODS: IFN-gamma production was detected based on the difference in supernatants between non-stimulated and stimulated peripheral blood mononuclear cells by phytohemagglutinin in 83 tubercular patients and 47 patients with pneumonia. Presence of a cavity on chest radiography and co-morbidities of pneumoconiosis, bronchiectasis, liver cirrhosis, renal failure on hemodialysis, diabetes mellitus (DM) and lung cancer were recorded for analysis. RESULTS: Interferon-gamma response, DM and a cavity on chest radiography were independent factors for predicting active pulmonary tuberculosis. Interferon-gamma response was decreased in patients with pulmonary tuberculosis compared with that in patients with non-tubercular pneumonia. Notably, M. tuberculosis infection was the principal factor correlated with IFN-gamma response. CONCLUSION: The IFN-gamma response was principally affected by M. tuberculosis infection and not by other co-morbidities. Further study is required to identify the mechanism of decreased IFN-gamma production.

Antioxidant properties of water extracts from Cassia tora L. in relation to the degree of roasting.

The antioxidant properties of water extracts from Cassia tora L. (WECT) prepared under different degrees of roasting were investigated. The water extracts of unroasted C. tora L. (WEUCT) showed 94% inhibition of peroxidation of linoleic acid at a dose of 0.2 mg/mL, which was higher than that of alpha-tocopherol (82%). Water extracts prepared from C. tora L. roasted at 175 degrees C for 5 min and at 200 degrees C for 5 min exhibited 83% and 82%, respectively, inhibition of linoleic acid peroxidation. This result indicated that the antioxidant activities of WECT decreased with longer roasting time or higher roasting temperature. The IC(50) of WEUCT in liposome oxidation induced by the Fenton reaction was 0.41 mg/mL, which was higher than that of alpha-tocopherol (IC(50) = 0.55 mg/mL). WEUCT also exhibited good antioxidant activity in enzymatic and nonenzymatic microsome oxidative systems. The water extracts of roasted C. tora L. increased in the degree of browning and produced chemiluminescence when compared with the unroasted sample. However, the total polyphenolic compounds of WECT decreased after the roasting process finished. In conclusion, the decrease in the antioxidant activity of water extracts from roasted C. tora L. might have been due to the degradation of Maillard reaction products and the decrease of polyphenolic compounds.

Identification and cloning of an Erwinia carotovora subsp. carotovora bacteriocin regulator gene by insertional mutagenesis.

Avirulent Erwinia carotovora subsp. carotovora CGE234-M403 produces two types of bacteriocin. For the purpose of cloning the bacteriocin genes of strain CGE234M403, a spontaneous rifampin-resistant mutant of this strain, M-rif-11-2, was isolated. By Tn5 insertional mutagenesis using M-rif-11-2, a mutant, TM01A01, which produces the high-molecular-weight bacteriocin but not the low-molecular-weight bacteriocin was obtained. By thermal asymmetric interlaced PCR, the DNA sequence from the Tn5 insertion site and the DNA sequence of a contiguous 1,280-bp region were determined. One complete open reading frame (ORF), designated ORF2, was identified within the sequenced fragment. The 3' end of another ORF, ORF1, was located upstream of ORF2. A noncoding region and a putative promoter were located between ORF1 and ORF2. Downstream from ORF2, the 5' end of another ORF (ORF3) was found. Deduction from the nucleotide sequence indicated that ORF2 encodes a protein of 99 amino acids, which showed high homology with Yersinia enterocolitica Yrp, a regulator of enterotoxin (Y-ST) production; Escherichia coli host factor 1, required for Qbeta-replicase; and Azorhizobium caulinodans NrfA, required for the expression of nifA. ORF2 was designated brg, bacteriocin regulator gene. A fragment containing ORF2 and its promoter was amplified and cloned into pBR322 and pHSG415r, and the recombinant plasmids, pBYL1 and pHYL1, were transferred into E. coli DH5. Plasmid pBYL1 was reisolated and transferred into the insertion mutant TM01A01. Transformants carrying the plasmid, which was reisolated and designated pBYL1, re-produced the low-molecular-weight bacteriocin.