Journal: Diagnostics

Article Title: Combined Immunofluorescence (IFA) and Fluorescence In Situ Hybridization (FISH) Assays for Diagnosing Babesiosis in Patients from the USA, Europe and Australia

doi: 10.3390/diagnostics10100761

Figure Lengend Snippet: IFA test results with the 36 Babesia genus FISH test positive samples from the USA.

Article Snippet: Smears on microscope slides prepared from the blood of hamsters infected with B. duncani (ATCC PRA-302) and B. microti (ATCC 30221D), provided by Dr. Alan Ashbaugh, University of Cincinnati, OH, USA, were used in IFA assays for detecting antibodies in clinical sera essentially as previously described [ , ] but with modifications for separately detecting IgM and IgG antibodies.

Techniques:

Journal: Diagnostics

Article Title: Combined Immunofluorescence (IFA) and Fluorescence In Situ Hybridization (FISH) Assays for Diagnosing Babesiosis in Patients from the USA, Europe and Australia

doi: 10.3390/diagnostics10100761

Figure Lengend Snippet: Comparison of diagnostic findings for babesiosis in Australia, Europe and the USA.

Article Snippet: Smears on microscope slides prepared from the blood of hamsters infected with B. duncani (ATCC PRA-302) and B. microti (ATCC 30221D), provided by Dr. Alan Ashbaugh, University of Cincinnati, OH, USA, were used in IFA assays for detecting antibodies in clinical sera essentially as previously described [ , ] but with modifications for separately detecting IgM and IgG antibodies.

Techniques: Comparison, Diagnostic Assay

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Knockout of microRNA-21 attenuates alcoholic hepatitis through the VHL/NF-κB signaling pathway in hepatic stellate cells

doi: 10.1152/ajpgi.00111.2018

Figure Lengend Snippet: Aberrant expressed inflammatory cytokines in the livers from heavy alcohol drinkers with steatohepatitis. A and B: total RNA was isolated from steatohepatitis patients with heavy alcohol consumption compared with healthy controls, and real-time PCR analysis was performed as described in materials and methods. The mRNA expression of inflammation markers [IL-6, monocyte chemoattractant protein-1 (MCP-1), and IL-1β; A] and cytokine (TNFα; B) was increased in steatohepatitis patients compared with healthy controls. C: increased expression of LPS receptor Toll-like receptor-4 (TLR4) in heavy alcohol consumers with steatohepatitis was verified by immunohistochemistry analysis compared with healthy controls (original magnification: ×20; scale bar = 100 μm). D: total RNA was collected from hepatocytes (CK8), cholangiocytes (CK19), hepatic stellate cells (HSCs; desmin), and macrophages (F4/80) isolated from control and ethanol-treated mice liver by laser capture microdissection using specific markers, and Taqman real-time PCR assay was carried out to detect miR-21 expression. Ethanol feeding significantly increased the hepatic expression of miR-21 in hepatocytes, cholangiocytes, HSCs, and macrophages when compared with control mice. *P < 0.05 vs. healthy controls.

Article Snippet: SuperArray quantitative PCR assay and qPCR analysis.

Techniques: Isolation, Real-time Polymerase Chain Reaction, Expressing, Immunohistochemistry, Laser Capture Microdissection

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Knockout of microRNA-21 attenuates alcoholic hepatitis through the VHL/NF-κB signaling pathway in hepatic stellate cells

doi: 10.1152/ajpgi.00111.2018

Figure Lengend Snippet: microRNA-21 (miR-21) depletion attenuates alcoholic liver injury in ethanol-fed mice. A: miR-21 expression was assessed by Taqman real-time PCR assay in wild-type (WT) and miR-21 KO mice with ethanol treatment relative to controls (n = 5). Ethanol feeding significantly increased the hepatic expression of miR-21 compared with WT control mice. Furthermore, miR-21 expression was significantly decreased in miR-21 KO (miR-21−/−) mice fed EtOH compared with WT EtOH-fed mice. B:iIn EtOH-fed miR-21−/− mice there was decreased serum ALT levels compared with WT EtOH-fed mice. C: in EtOH-fed mice, liver cell swelling, irregular nucleus size, shrinkage in varying degrees, and fatty degeneration of a large number of liver cells were all observed. D: pathological changes and scores were reduced in EtOH-fed miR-21−/− mice compared with WT EtOH-fed mice. The results shown represent the means ± SE from 4 independent experiments. *P < 0.05, relative to controls. #P < 0.05, relative to EtOH-fed mice.

Article Snippet: SuperArray quantitative PCR assay and qPCR analysis.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Knockout of microRNA-21 attenuates alcoholic hepatitis through the VHL/NF-κB signaling pathway in hepatic stellate cells

doi: 10.1152/ajpgi.00111.2018

Figure Lengend Snippet: microRNA-21 (miR-21) modulates NF-κB signaling pathway in LPS-activated hepatic stellate cells (HSCs) in vitro. A and B: expression levels of key mediators of NF-κB signaling pathway are altered in anti-miR-21-treated HSCs after LPS stimulation. Relative gene expression profile between anti-miR-21-treated HSCs after LPS stimulation vs. anti-miR controls is shown. The expression of a panel of diverse inflammation-associated genes was evaluated by real-time PCR assay using Human Chemokines & Receptors PCR Array (PAHS-022 from SABiosciences), which was selected based on Ingenuity Pathway Analysis with the focus of NF-κB signaling associated gene list). A: gene expression relative to GAPDH was plotted as the Volcano Plots, depicting the relative expression levels (Log10) for selected genes in anti-miR-Con vs. anti-miR-21. B: the relative expression levels and P values for each gene in the related samples were also plotted against each other in the scatterplot. The key mediators of NF-κB signaling pathway, NF-κB1, Toll-like receptor-4 (TLR4), MYD88, and von Hippel-Lindau (VHL) are the most altered genes in anti-miR-21-treated HSCs after LPS stimulation. Data represent mean from 3 separate experiments. C: Ingenuity Pathway Analysis based on PCR array discoveries showed that miR-21 may target VHL and subsequently alter the NF-κB signaling pathway.

Article Snippet: SuperArray quantitative PCR assay and qPCR analysis.

Techniques: In Vitro, Expressing, Real-time Polymerase Chain Reaction