Effect of Fermented Sauropus Androgynus Leaves on Blood Lipid Fraction and Haematological Profile in Broiler Chickens

This study was conducted to evaluate effect of fermented Sauropus androgynus leaves on blood lipid fractions and haematological profiles in broilers. One hundred and twelve broilers were distributed to 7 treatment groups. One group was fed diets without Sauropus androgynus leaves as the control, and other six groups were fed Sauropus androgynus leaves fermented by Neurospora crassa, Lactobacillus sp. or Saccharomyces cerevisiae at level of 25 g or 50 g/kg diet. Experimental results showed that the treatments had no effect on cholesterol, high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein cholesterol (LDL-c) and atherogenic index, very low-density lipoprotein cholesterol (VLDL-c) and triglyceride concentration (P>0.05). It was shown that fermented Sauropus androgynus leaves significantly affected red blood count (RBC), white blood count (WBC), packed cell volume (PCV), trombosit dan erythrocyte sedimentation rate (ESR) (

Seronegative, aviremic hemodialysis patients with HCV-specific T cell responses.

*<p>N.D., not determined.</p

HCV-specific T cell responses in seronegative, aviremic hemodialysis patients.

<p><b>(A)</b> Direct <i>ex vivo</i> IFN-γ ELISpot assay was performed to examine HCV-specific T cell responses in PBMCs of 76 hemodialysis patients. Five patients were seropositive with viremia (left side), and 71 patients were seronegative and aviremic (right side). PBMCs were stimulated with the 49 matrix mixes of 600 overlapping peptides covering the complete HCV polyprotein sequence (Materials and Methods and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0062319#pone.0062319.s001" target="_blank">Figure S1A</a>). The sum of A-X mixes and the sum of AA-YY mixes were calculated respectively, and their average value was presented for total HCV-specific T cell responses in each patient. Box with dashed lines denotes seronegative, aviremic patients with obvious T cell responses. <b>(B)</b> In the eight seronegative, aviremic patients with obvious T cell responses (CMI-1 to CMI-8), the results of IFN-γ ELISpot assay were presented by each peptide mix (A to X). Relationship between peptide mixes and HCV proteins are also displayed. <b>(C)</b> IFN-γ ICS was performed to identify the T cell subset responding to HCV epitopes. PBMCs were stimulated with epitope peptides identified in the IFN-γ ELISpot assay in the each patient. FACS dot plots are representative data from two patients, CMI-1 and CMI-4.</p

Polyfunctionality of HCV-specific T cells in seronegative, aviremic hemodialysis patients.

<p>PBMCs were stimulated with an epitope peptide or a peptide mix, and production of IFN-γ, TNF-α, IL-2, and MIP-1β was simultaneously assessed in multi-cytokine ICS in order to evaluate polyfunctionality of HCV-specific CD4⁺ T cells. In each patient, the fraction of T cells positive for a given number of functions is presented as bar graphs. In the patient with CD8⁺ T cell responses (CMI-4), cytotoxic degranulation function was also evaluated by CD107a staining in addition to multi-cytokine ICS for IFN-γ, TNF-α, IL-2, and MIP-1β.</p

Cytokine profile of HCV-specific T cells in seronegative, aviremic hemodialysis patients.

<p><b>(A)</b> After stimulating PBMCs with an epitope peptide or a peptide mix for 3 days, culture supernatant was harvested, and the concentrations of IFN-γ and TNF-α determined in the culture supernatant by CBA. There are two groups of patients in terms of cytokine profile: group I, patients with IFN-γ and TNF-α responses; group II, patients with TNF-α-predominant responses. <b>(B)</b> TNF-α ICS was performed in the patients of group II. FACS dot plots are representative data from two patients, CMI-7 and CMI-8.</p

Hemodialysis patients with secondary nested RT-PCR (+).

*<p>N.D., not determined.</p

A novel multifunctional peptide oligomer of bacitracin with possible bioindustrial and therapeutic applications from a Korean food-source <i>Bacillus</i> strain - Fig 1

<p>(A) A neighbor-joining tree based on nearly complete 16S rRNA gene sequences showing relations between strain CS32 and related members of the genus <i>Bacillus</i>. The percentages at the nodes are the levels of bootstrap support based on neighbor-joining analyses of 1000 resampled datasets. The sequence of <i>Bacillus licheniformis</i> ATCC 14580(T) served as an outgroup. The scale bar: 0.01 nucleotide substitutions per position. (B) Production of CSP32 and the growth-inhibitory activity against <i>Mycobacterium smegmatis</i> ATCC 9341. The antibacterial activity is maximal at 60 h. The zone of inhibition against <i>Mycobacterium smegmatis</i> ATCC 9341 at every 12 h is shown. Cultivation was carried out in 250-mL flasks with 50 mL of a medium, at pH 7 and 37°C, with shaking at 180 rpm.</p

Effects of CSP32 on NO production and expression of inflammation-associated proteins in LPS-stimulated RAW 264.7 macrophage cells.

<p>The cells were treated with LPS (1 μg/mL) in the presence of various concentrations of CSP32 (10, 50, or 100 μg/mL) at 37°C for 24 h. (A) The amounts of NO were determined using the Griess reagent in the culture medium (Top panel). Equal amounts of cell lysates were resolved on SDS polyacrylamide gels, transferred to PVDF membranes, and probed with antibodies against iNOS and COX-2. β-Actin served as an internal control for western blot analysis (bottom panel). Effects of CSP32 on LPS-induced mRNA expression of iNOS, COX-2, TNF-α, IL-1β, and IL-6. (B) After LPS treatment for 2–12 h, the levels of iNOS, COX-2, TNF-α, IL-1β, and IL-6 mRNA were determined by RT-PCR. <i>GAPDH</i> served as an internal control for RT-PCR assays. Suppressive effects of CSP32 on TNF-α, IL-1β, and IL-6 production in LPS-stimulated RAW 264.7 macrophage cells. (C) The cells were incubated with the indicated concentrations of CSP32 for 30 min before treatment with LPS (1 μg/mL) for 24 h. After incubation for 24 h, the supernatant was collected, and the amounts of these proinflammatory cytokines were measured by ELISAs. Results represent the mean±S.D of three independent experiments performed in triplicate. * p<0.01; compared to LPS alone.</p

Comparison of MS data with predicted molecular weight of each subunit.

<p>Comparison of MS data with predicted molecular weight of each subunit.</p

Comparative analysis of N-terminal amino acid sequences.

<p>Comparative analysis of N-terminal amino acid sequences.</p