Journal: Advanced Science

Article Title: Palmitoyltransferase ZDHHC3 Aggravates Nonalcoholic Steatohepatitis by Targeting S ‐Palmitoylated IRHOM2

doi: 10.1002/advs.202302130

Figure Lengend Snippet: Hepatocyte‐specific loss of Zdhhc3 protects against HFHC‐induced NASH pathogenesis. a‐g) Records for the body weight (a), liver weight and the ratio of liver weight/body weight (%) (LW/BW) (b), fasting blood glucose levels (c), fasting insulin levels (d), HOMA‐IR index (e), glucose tolerance test (GTT) (f) and liver TG, NEFA and TC contents (g) of the HFHC‐fed Zdhhc3 ‐Flox mice and Zdhhc3 ‐HepKO mice; NCD diet‐fed mice were treated as corresponding control ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). h) Pearson analysis indicating the correlations between fasting insulin, ratio of liver weight/body weight (%), and liver TG contents in indicated groups ( n = 10 per parameter; P < 0.001 for all of these correlations). i‐k) Representative pictures of H&E staining, oil red O staining, histological NAS score (i) changes, sirius red staining, masson staining (j), and F4/80 and CD11b positive cells expression (k) in indicated groups (magnification, 100× for H&E staining, oil red O staining, masson staining and sirius red staining; magnification, 200× for F4/80 and CD11b staining; n = 10 images per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). l, Records for inflammation‐related cytokines profiles including IL‐6, TNF‐α, IL‐1β, IL‐18 and CCL2 in serum from HFHC‐fed HepKO or Flox mice ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). m, Records for liver function‐related indicators including ALT, AST, AKP, and GGT in serum from HFHC‐fed HepKO or Flox mice ( n = 10 mice per group; P < 0.05 versus Zdhhc3 ‐Flox HFHC group). Data are expressed as mean ± SEM. The relevant experiments presented in this part were performed independently at least three times. Significance is determined by two‐way analysis of variance (ANOVA) followed by multiple comparisons test analysis (a‐g) or 2‐sided Student's t ‐test (i‐m). The P ‐value < 0.05 was considered as significant difference.

Article Snippet: The TNF‐α (Cat: ab100747), IL‐1β (Cat: ab197742), IL‐6 (Cat: ab222503), CCL2 (Cat: ab208979), IL‐18 (Cat: ab216165) and insulin (Cat: ab285341) ELISA kits were purchased from Abcam.

Techniques: Control, Staining, Expressing

Journal: Journal of aging science

Article Title: Angiotensin Converting Enzyme 2 (ACE2) Expression in the Aged Brain and Visual System

doi:

Figure Lengend Snippet: Cell- and tissue-specific patterns of ACE2 expression (mRNA) in human ocular cells and in selected areas of the brain involved in visual processing. The extent of ACE2 transmembrane receptor expression in the plasma membrane is an important indicator of ACE2 gene function, the susceptibility to SARS-CoV-2 invasion and the development of COVID-19. Significant ACE2 receptor expression was initially observed in the whole brain, whole eye and whole retina and subsequently cells and tissues involved in visual signal acquisition and processing were analyzed for ACE2 abundance. ACE2 acts as a receptor for the spike (S1) glycoprotein of the human coronavirus HCoV-NL63 and the human SARS-CoV and SARS-CoV-2 virus that causes COVID-19 (see: https://www.ncbi.nlm.nih.gov/gene?Db=gene&Cmd=DetailsSearch&Term=59272 ; last accessed 9 September 2021); in the mRNA experiments both G3PDH and β-actin DNA probes were used as abundance controls; in this experiment relative signal strength refers to control β-actin mRNA levels in the same tissues ; in this and one previous study the highest expression of the SARS-CoV-2 ACE-2 receptor was found in the cerebral cortex and the occipital lobe, the pons and medulla oblongata of the brain and in the whole retina, the optic nerve and the ocular choroid and RPE cells of the eye. High expression of the ACE2 receptor in the eye and neural pathways involved in vision and visual processing may predispose this circuitry of the visual system to attracting SARSCoV-2 and viral invasion. A vertical dashed line at 1.0 is included for ease of comparison; a minimum of N=3 HyBond N+ or MTE filters were used for each tissue determination; *p<0.05; **p<0.01 (ANOVA); error bars represent one standard deviation of the mean.

Article Snippet: Protein extracts from cells and/or tissues involved in human vision, including ocular choroidal fibroblasts, trabecular meshwork cells, non-pigmented ciliary epithelial cells, retinal pigment epithelial cells, corneal epithelial cells, whole retina, whole eye and tissues from brain visual processing pathways including the optic nerve, cerebellum, pons, temporal lobe, occipital lobe (Brodmann Area 17, containing the primary visual cortex), cerebral cortex and whole brain were generated using a ProteoExtract Complete Mammalian Proteome Extraction Kit (cat no. 539779, Calbiochem/Millipore-Sigma Burlington MA) and were assayed for protein concentration using a Non-Interfering Protein Assay kit (cat no. 488250, Calbiochem/Millipore-Sigma) at 480 nm; protein samples were stored in at −81°C according to the manufacturers protocol (Millipore-Sigma); ACE2 protein [UniProtKB - Q9BYF1 (ACE2_HUMAN)] abundance these selective cell extracts of the visual system and brain were analyzed using a quantitative colorimetric (450 nm) sandwich ELISA specific for human ACE2 using a Fluoroskan Ascent FT Microplate Fluorometer and Luminometer (Cat no. 5200220, ThermoFisher Scientific, Waltham MA; sensitivity 1052 pg/ml; detection range 1.5 ng/ml - 255 ng/ml (Human ACE2 ELISA Kit ab235649; Abcam Cambridge MA, USA); human recombinant ACE2 protein (Abcam ab151852) and human beta-actin (β-actin; anti-beta actin antibody (Abcam ab8227) were used as internal controls to quantify relative ACE2 and β-actin protein abundance in each sample according to the manufacturer’s instructions ( ); concentrations of ACE2 were measured in triplicate and interpolated from the ACE2 standard curve and corrected for sample dilution as according to manufacturer’s protocol; human ACE2 was expressed as ng/ml (Abcam; https://www.abcam.com/human-ace2-elisa-kit-ab235649.html ; last accessed 9 September 2021; see ).

Techniques: Expressing, Standard Deviation

Journal: Journal of aging science

Article Title: Angiotensin Converting Enzyme 2 (ACE2) Expression in the Aged Brain and Visual System

doi:

Figure Lengend Snippet: Cell- and tissue-specific patterns of ACE2 expression at the protein level in selected human ocular cells and in anatomical areas of the brain involved in visual processing; bar graph of ELISA analysis (see ); we observed a very strong correlation between ACE2 mRNA abundance with ACE2 protein abundance ; the highest expression of the SARS-CoV-2 receptor ACE-2 protein was found in RPE cells, the whole retina, optic nerve, the pons and the occipital lobe that contains the primary visual cortex and main visual processing area (Brodmann Area17); relative signal strength refers to control β-actin protein levels in the same tissues; see text for further details; a vertical dashed line at 1.0 is included for ease of comparison; a minimum of N=3 ELISA analyses were performed for each protein determination in cells or tissues; *p<0.05; **p<0.01 (ANOVA); error bars represent one standard deviation of the mean.

Article Snippet: Protein extracts from cells and/or tissues involved in human vision, including ocular choroidal fibroblasts, trabecular meshwork cells, non-pigmented ciliary epithelial cells, retinal pigment epithelial cells, corneal epithelial cells, whole retina, whole eye and tissues from brain visual processing pathways including the optic nerve, cerebellum, pons, temporal lobe, occipital lobe (Brodmann Area 17, containing the primary visual cortex), cerebral cortex and whole brain were generated using a ProteoExtract Complete Mammalian Proteome Extraction Kit (cat no. 539779, Calbiochem/Millipore-Sigma Burlington MA) and were assayed for protein concentration using a Non-Interfering Protein Assay kit (cat no. 488250, Calbiochem/Millipore-Sigma) at 480 nm; protein samples were stored in at −81°C according to the manufacturers protocol (Millipore-Sigma); ACE2 protein [UniProtKB - Q9BYF1 (ACE2_HUMAN)] abundance these selective cell extracts of the visual system and brain were analyzed using a quantitative colorimetric (450 nm) sandwich ELISA specific for human ACE2 using a Fluoroskan Ascent FT Microplate Fluorometer and Luminometer (Cat no. 5200220, ThermoFisher Scientific, Waltham MA; sensitivity 1052 pg/ml; detection range 1.5 ng/ml - 255 ng/ml (Human ACE2 ELISA Kit ab235649; Abcam Cambridge MA, USA); human recombinant ACE2 protein (Abcam ab151852) and human beta-actin (β-actin; anti-beta actin antibody (Abcam ab8227) were used as internal controls to quantify relative ACE2 and β-actin protein abundance in each sample according to the manufacturer’s instructions ( ); concentrations of ACE2 were measured in triplicate and interpolated from the ACE2 standard curve and corrected for sample dilution as according to manufacturer’s protocol; human ACE2 was expressed as ng/ml (Abcam; https://www.abcam.com/human-ace2-elisa-kit-ab235649.html ; last accessed 9 September 2021; see ).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Frontiers in Veterinary Science

Article Title: One Health in Action: Operational Aspects of an Integrated Surveillance System for Zoonoses in Western Kenya

doi: 10.3389/fvets.2019.00252

Figure Lengend Snippet: Biological samples collected, laboratory tests performed, and pathogens investigated, during an integrated surveillance program for zoonoses in western Kenya.

Article Snippet: , , Toxoplasma spp . (Anti-Toxoplasma gondii IgG Human ELISA Kit [Abcam] and Toxoplasma IgM ELISA [DRG International]) , As per manufacturer's instructions.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Stripping Membranes, Microscopy