Journal: Asian Journal of Andrology

Article Title: The identification of AMZ2 as a candidate causative gene in a severe teratozoospermia patient characterized by vacuolated spermatozoa

doi: 10.4103/aja202321

Figure Lengend Snippet: Identification of a homozygous AMZ2 variant in an individual with teratozoospermia from a consanguineous family. ( a ) Diff-Quik staining of human spermatozoa obtained from one individual with an AMZ2 variant and one healthy control. Most spermatozoa from the individual with the AMZ2 variant showed sperm-head vacuolation. Sperm-head vacuoles are indicated with white triangles. ( b ) Validation of the AMZ2 c.520A>G (p.Thr174Ala) variant in the proband’s pedigree using Sanger sequencing. The mutation (M) was homozygous in the proband (IV-1) and both parents were heterozygous carriers. The double horizontal line indicates the consanguineous union. Squares and circles denote male and female family members, respectively. The proband is indicated with an arrow. I–V represent five generations from the oldest to the youngest. AMZ2 : archaelysin family metallopeptidase 2; M: mutation; WT: wild-type.

Article Snippet: The membrane was blocked in 5% nonfat milk for 1 h at room temperature (RT), then incubated overnight at 4°C with AMZ2 mouse monoclonal antibody (1:100, sc-365345, Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Variant Assay, Diff-Quik, Staining, Control, Biomarker Discovery, Sequencing, Mutagenesis

Journal: Asian Journal of Andrology

Article Title: The identification of AMZ2 as a candidate causative gene in a severe teratozoospermia patient characterized by vacuolated spermatozoa

doi: 10.4103/aja202321

Figure Lengend Snippet: Archaelysin family metallopeptidase 2 (AMZ2) variant data for the patient analyzed in this study

Article Snippet: The membrane was blocked in 5% nonfat milk for 1 h at room temperature (RT), then incubated overnight at 4°C with AMZ2 mouse monoclonal antibody (1:100, sc-365345, Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Variant Assay, Mutagenesis

Journal: Asian Journal of Andrology

Article Title: The identification of AMZ2 as a candidate causative gene in a severe teratozoospermia patient characterized by vacuolated spermatozoa

doi: 10.4103/aja202321

Figure Lengend Snippet: Effects of the AMZ2 missense mutation on protein level. ( a ) Structural model of wild-type AMZ2 protein (in grey) and the mutant p.Thr (in green) 174Ala (in magenta). Cyan indicates the zinc-binding motif. Dashed lines represent hydrogen bonds. ( b ) Western blot analysis of AMZ2 levels in human spermatozoa from the individual with the AMZ2 variant and a healthy control. AMZ2 was absent in the spermatozoa of the individual with the AMZ2 variant. GAPDH was used as the loading control. ALA: alanine; AMZ2 : archaelysin family metallopeptidase 2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ILE: isoleucine; M: mutation; PHE: phenylalanine; THR: threonine; WT: wild-type.

Article Snippet: The membrane was blocked in 5% nonfat milk for 1 h at room temperature (RT), then incubated overnight at 4°C with AMZ2 mouse monoclonal antibody (1:100, sc-365345, Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Techniques: Mutagenesis, Binding Assay, Western Blot, Variant Assay, Control

Journal: Antioxidants

Article Title: Lindera obtusiloba Attenuates Oxidative Stress and Airway Inflammation in a Murine Model of Ovalbumin-Challenged Asthma

doi: 10.3390/antiox9070563

Figure Lengend Snippet: Effects of LOL on inflammatory cell number, Th2 cytokines, eotaxin, MUC5AC, total IgE and OVA-specific IgE in OVA-challenged asthma model. ( A ) The inflammatory cells were attached on slide and stained with Diff-Quik stain reagent. ( B ) The levels of IL-4, IL-5, IL-13, eotaxin, and MUC5AC in the BALF were determined by ELISA. ( C ) The total IgE and OVA-specific IgE in the serum were determined using ELISA. NC: normal control mice; OVA: OVA-challenged mice; DEX: dexamethasone (3 mg/kg) + OVA-challenged mice; LOL: LOL (50 or 100 mg/kg) + OVA-challenged mice. The values are expressed as the means ± SD ( n = 7/group). # p < 0.01, significantly different from NC group; †† p < 0.01, significantly different from OVA-challenged group.

Article Snippet: The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-4, IL-5, IL-6, IL-13, and eotaxin (R&D system, Minneapolis, MN, USA), MUC5AC (Cusabio Biotech Co. Wuhan, China) and IgE (BioLegend, CA, USA) were used according to the manufacturer’s instructions.

Techniques: Staining, Diff-Quik, Enzyme-linked Immunosorbent Assay, Control

Journal: Antioxidants

Article Title: Lindera obtusiloba Attenuates Oxidative Stress and Airway Inflammation in a Murine Model of Ovalbumin-Challenged Asthma

doi: 10.3390/antiox9070563

Figure Lengend Snippet: Effects of LOL on activation of MAPKs/AP-1, p65NF-κB and expression of MUC5AC in OVA-challenged asthma model. The protein levels of ( A – E ) MAPKs/AP-1, ( F ) p65NF-κB and ( G ) MUC5AC in the lung tissues were determined by western blot analysis. β-actin was used to confirm equal protein loading. NC: normal control mice; OVA: OVA-challenged asthma mice; DEX: dexamethasone (3 mg/kg) + OVA-challenged asthma mice; LOL: LOL (50 or 100 mg/kg) + OVA-challenged asthma mice. The values are expressed as the means ± SD ( n = 7/group). # p < 0.01, significantly different from NC group; †† p < 0.01, significantly different from OVA-challenged group.

Article Snippet: The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-4, IL-5, IL-6, IL-13, and eotaxin (R&D system, Minneapolis, MN, USA), MUC5AC (Cusabio Biotech Co. Wuhan, China) and IgE (BioLegend, CA, USA) were used according to the manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Western Blot, Control