Mempelajari Sifat Fisika Sol Karet Cetak Dengan Filler Cangkang Telur Ayam

Tujuan penelitian adalah untuk menpelajari sifat fisika sol karet cetak dengan filler cangkang telur ayam. Sifat fisika yang dipelajari meliputi kekerasan, tegangan putus, ketahanan sobek dan ketahanan kikis. Penelitian dilakukan dengan 4 tahap yaitu pembuatan filler cangkang telur ayam, pembuatan sol karet cetak, pengujian sifat fisika dan penilaian secara visual. Perlakuan terdiri dari penggunaan cangkang telur ayam menggantikan filer karbon hitam meliputi perlakuan tanpa penggunaan cangkang telur ayam (A1), penggunaan filler cangkang telur ayam 15 Phr (B1), penggunaan filer cangkang telur ayam 30 Phr (C1) dan penggunaan filler cangkang telur ayam 45 Phr (D1). Hasil penelitian menunjukkan bahwa cangkang telur ayam dapat digunakan sebagai filler pada pembuatan sol karet cetak. Penggunaan filler cangkang telur ayam yang semakin meningkat menghasilkan sol karet cetak dengan kekerasan yang cenderung semakin menurun, tegangan putus yang semakin menurun, ketahanan sobek yang semakin menurun dan ketahanan kikis yang semakin meningkat. Secara fisual sol karet cetak yang dihasilkan dari filler cangkang telur ayam menghasilkan sol karet cetak yang baik (tidak cacat berupa sobek, lubang, lepuh, retak dan goresan)

miR21 mediated sinusoidal endothelial dysfunction is the early event in NASH progression.

<p><i>A</i> and <i>B</i>. Immunofluorescence images for localization of sinusoidal endothelial dysfunction biomarkers (VEGFR-2, ICAM-1 and E-selectin) from liver sections of toxin model groups (fig. A) DIO+ BDCM (1w) and miR21 KO+BDCM (1w) and dietary model (fig. B) MCD (4w) and miR21 KO+MCD (4w). Immunoreactivity with red dots shows the localization of SED biomarkers.</p

Upregulation of miR21 and Repression of Grhl3 by Leptin Mediates Sinusoidal Endothelial Injury in Experimental Nonalcoholic Steatohepatitis

<div><p>Sinusoidal endothelial dysfunction (SED) has been found to be an early event in nonalcoholic steatohepatitis (NASH) progression but the molecular mechanisms underlying its causation remains elusive. We hypothesized that adipokine leptin worsens sinusoidal injury by decreasing functionally active nitric oxide synthase 3 (NOS)3 via miR21. Using rodent models of NASH, and transgenic mice lacking leptin and leptin receptor, results showed that hyperleptinemia caused a 4–5 fold upregulation of hepatic miR21 as assessed by qRTPCR. The upregulation of miR21 led to a time-dependent repression of its target protein Grhl3 levels as shown by western blot analyses. NOS3-p/NOS3 ratio which is controlled by Grhl3 was significantly decreased in NASH models. SED markers ICAM-1, VEGFR-2, and E-selectin as assessed by immunofluorescence microscopy were significantly up regulated in the progressive phases of NASH. Lack of leptin or its receptor in vivo, reversed the upregulation of miR21 and restored the levels of Grhl3 and NOS3-p/NOS3 ratio coupled with decreased SED dysfunction markers. Interestingly, leptin supplementation in mice lacking leptin, significantly enhanced miR21 levels, decreased Grhl3 repression and NOS3 phosphorylation. Leptin supplementation in isolated primary endothelial cells, Kupffer cells and stellate cells showed increased mir21 expression in stellate cells while sinusoidal injury was significantly higher in all cell types. Finally miR21 KO mice showed increased NOS3-p/NOS3 ratio and reversed SED markers in the rodent models of NASH. The experimental results described here show a close association of leptin-induced miR21 in aiding sinusoidal injury in NASH.</p></div

Sinusoidal endothelial dysfunction (SED) in NASH progression is mediated by leptin.

<p><i>A</i>. mRNA expression of sinusoidal endothelial dysfunction biomarkers (VEGFR-2, ICAM-1, E-selectin, VCAM-1, Cadherin 5 (Cdh5), VEGF-α and CD34) as measured by quantitative real-time PCR in DIO+BDCM (1w), ob/ob gene deleted mice (Lep KO (1w)) and db/db gene deleted mice (Lepr KO (1w)). Y-axis shows fold of mRNA expression of SED biomarkers normalized against DIO only groups. <i>B</i>. Immunofluorescence images for localization of SED biomarkers (VEGFR-2, ICAM-1 and E-selectin) from liver sections of both toxin model (DIO, DIO+BDCM (1w), Lep KO (1w), Lepr KO (1w)) and dietary model (MCS (4w) and MCD (4w)) of NASH.</p

miR21 play a key role in leptin signaling of sinusoidal endothelial dysfunction.

<p><i>A</i> and <i>C</i>. Phosphorylation of endothelial nitric oxide synthase (NOS3) is the key event in endothelial function. To access levels of NOS3 phosphorylation in liver homogenate, western blot analysis was carried out for phosphorylated NOS3 (NOS3-p) and native NOS3 protein. The mice groups for toxin model (fig. A) are DIO+BDCM (1w), DIO+BDCM (4w) and 3 individual miR21 KO mice (M1, M2 and M3) and the mice groups for dietary model (fig. C) are MCS (4w), miR21 KO fed with MCS diet (miR21 KO+MCS (4w)), MCD (4w) and miR21 KO mice fed with MCD diet (miR21 KO+MCD (4w)). The corresponding β-actin levels are shown in the lower panel. <i>B</i> and <i>D</i>. Levels of phosphorylated NOS3 (NOS3-p) protein normalized against respective NOS3 levels and β-actin levels (NOS3-p/NOS3 ratio) were plotted. Y-axis represent arbitrary unit of NOS3-p/NOS3 ratio of mice groups from both BDCM (fig. B) and diet model (fig. D) of NASH. P<0.05 is considered statistically significant (*).</p

Increased hepatic leptin is associated with NASH progression in obesity.

<p>qRTPCR analysis of hepatic leptin mRNA expression in two toxin model of NASH. <i>A</i>. Bromodichloromethane (BDCM) model: Y-axis represents fold of leptin mRNA expression in DIO, DIO mice exposed with BDCM for 24h, for 48h, for 1week and for 4 weeks post BDCM exposure. <i>B</i>. Carbon tetrachloride (CCl₄) model: Y-axis represents fold of leptin mRNA expression in DIO and DIO mice exposed with CCl₄ for 1w. n = 3, P<0.05 is considered statistically significant (*). <i>C</i>. Picro-sirius red (PSR) staining of liver sections of DIO, DIO+BDCM at 24h, DIO+BDCM at 48h, DIO+BDCM at 1w and DIO+BDCM at 4w post BDCM exposure; 20× images (n = 3). Black arrowhead depicts macro and micro vesicular fibrosis. <i>D</i>. Stages of fibrosis of stained liver sections from two different model of NASH (toxin and dietary model) were reviewed using the criteria of the NIH Non Alcoholic Steatohepatitis Clinical Research Network (NIH NASH CRN). Table depicts the NASH CRN scores for DIO, DIO+BDCM (toxin model) and MCS, MCD (Dietary model).</p

Increased miR21 expression, subsequent decreased expression of Grhl3 protein and NOS3 phosphorylation is mediated by leptin.

<p><i>A</i>. miR21 expression as measured by quantitative real-time PCR in DIO mice exposed with BDCM (DIO+BDCM (1w)), ob/ob gene deficient mice exposed with BDCM (Lep KO (1w)) and ob/ob gene deficient mice supplemented with leptin exposed with BDCM (Lep KO+Leptin (1w)). P<0.05 is considered statistically significant (*) <i>B</i>. Western blot analysis of Grhl3 in liver homogenates of BDCM exposed DIO mice (DIO+BDCM (1w)), mice that lacked the ob/ob gene and exposed with BDCM (Lep KO (1w)) and Lep KO supplemented with leptin and exposed with BDCM (Lep KO+Leptin (1w)). <i>C</i>. Column graph depict the band quantification analysis of Grhl3 protein with corresponding β-actin as shown in lower levels of fig B. <i>D</i>. Western blot analysis of phosphorylated NOS3 (NOS3-p) and NOS3 in DIO+BDCM (1w), Lep KO (1w) and Lep KO+Leptin (1w) mice group. <i>E</i>. NOS3-p/NOS3 ratio (indication of vascular endothelium function) was plotted after band quantification and normalization against respective β-actin. Y-axis represent arbitrary unit of NOS3-p/NOS3 ratio of DIO+BDCM (1w), Lep KO (1w) and Lep KO+Leptin (1w) mice groups. P<0.05 is considered statistically significant (*). #Band image of DIO+BDCM (1w) has been cropped from the same immunoblot image and placed separately in both fig B and D due to presence of other mouse group in between the DIO+BDCM (1w) and Lep KO (1w) lane which is not explained in this manuscript. The cropped images of the blot are separated by a distinct blank space to show the non-continuity of the image.</p

GW chemical-induced altered gut microbiome modulates gap junction protein levels in the small intestine.

<p>A. Tissue levels of Claudin-2 in control (GW-Con), treated (GW-T) and antibiotic treated (GW-Ab) samples as observed by immunofluorescent microscopy after labelling the protein with red fluorescent secondary antibody. Nuclear staining is shown by DAPI (blue) stain. B. Tissue levels of Occludin in control (GW-Con), treated (GW-T) and antibiotic treated (GW-Ab) samples as observed by immunofluorescent microscopy after labelling the protein with red fluorescent secondary antibody. Nuclear staining is shown by DAPI (blue) stain. C. Morphometry of fluorescence intensity as observed in the region of interest (ROI) Claudin-2 immunoreactivity and D. occludin immunoreactivity. E. Western blot analysis of Claudin-2 and Occludin in small intestine tissue homogenate. F-G. morphometric analysis of western blot. *(p<0.05). Data is represented as Mean+/- SE.</p

Gulf war chemical exposure-induced changes in gut microbiome modulates neuroinflammation and intestinal cytokine release.

<p>A. Frontal cortex tissue slices were probed for IL-1β immunoreactivity in control (GW-Con, treated (GW-T), TLR4 knockout mice treated with GW Chemical exposure (GW-TLR4KO) and antibiotic treated (GW-Ab) groups. Specific immunoreactivity to IL-1β as evident by dark brown spots are indicated by black arrows and areas of interest are outlined by red circles. B. Frontal cortex tissue slices were probed for MCP-1 immunoreactivity in control (GW-Con, treated (GW-T), TLR4 knockout mice treated with GW Chemical exposure (GW-TLR4KO) and antibiotic treated (GW-Ab) groups. Specific immunoreactivity to MCP-1 as evident by dark brown stain/spots are indicated by black arrows and areas of interest are outlined by red circles. C. Small intestine tissue slices were probed for IL-1β immunoreactivity in control (GW-Con, treated (GW-T), TLR4 knockout mice treated with GW Chemical exposure (GW-TLR4KO) and antibiotic treated (GW-Ab) groups. D. Small intestine tissue slices were probed for MCP-1 immunoreactivity in control (GW-Con, treated (GW-T), TLR4 knockout mice treated with GW Chemical exposure (GW-TLR4KO) and antibiotic treated (GW-Ab) groups.</p

Schematic representation of dosage pattern:

<p><b>A.</b> dosage pattern of control group (GW-Con) n = 4, C57BL/6J mice were exposed to vehicle/solvent (Veh) of the Gulf war chemicals and antibiotics. <b>B.</b> C57BL/6J (n = 6) and TLR4 KO (n = 6) mice were exposed to gulf war chemicals Pyridostigmine bromide (Pb), Permethrin (Per) and Corticosterone (Cort). <b>C.</b> C57BL/6J (n = 6) mice were co-exposed to gulf war chemicals (Pb, Per and Cort) and Antibiotics (Ab).</p