Journal: Dermatologic Therapy

Article Title: Cutaneous manifestations following COVID‐19 vaccination: A report of 25 cases

doi: 10.1111/dth.15651

Figure Lengend Snippet: Characteristics of the patients

Article Snippet: 24 , Female , 32 , None , None , AstraZeneca , First , Urticarial and targetoid lesions , Fever, , 20 days , WBC:10100 AST:79 ALT:139 ESR:49 CRP: negative , Prednisolone, topical clobetasol , Significant improvement , None.

Techniques: Imaging, Injection

Journal: Frontiers in Allergy

Article Title: Successful Desensitization to mRNA COVID-19 Vaccine in a Case Series of Patients With a History of Anaphylaxis to the First Vaccine Dose

doi: 10.3389/falgy.2022.825164

Figure Lengend Snippet: Patient demographics, reaction to first vaccine dose, treatment of reaction, latency period between first dose, and allergist evaluation, result of PEG SPT and response to desensitization.

Article Snippet: 5 , F 57 , Caucasian , Bronchial Asthma Ehlers-Danlos syndrome Osteopenia , Septra , 20 min : facial numbness and swelling, cough and difficulty breathing, hoarseness, and chest pain , Negative , mRNA (Pfizer-BioNTech) , Third dose: tongue tingling. 1 week: maculopapular rash, desquamation of the skin. Treatment: desloratadine 10 mg PO, mometasone cream topical application..

Techniques: Recombinant

Journal: Human Molecular Genetics

Article Title: Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation

doi: 10.1093/hmg/ddy215

Figure Lengend Snippet: Interaction between cofilin-1 and pERK1/2. (A) Immunoblot showing interaction of cofilin-1 and pERK12 in IP experiments. Proteins extracted from C2C12 cells were subjected to IP using antibodies against cofilin-1, N-Wasp, ARP2 or profilin-1. Proteins in immunoprecipitates were separated by SDS-PAGE and IB using antibodies against pERK1/2, cofilin-1, N-Wasp, ARP2 or profilin-1. Immunoglobulin G (IgG) was used as a negative control. Representative from three independent repeats. (B) Immunoblot showing interaction of cofilin-1 and pERK12 in IP experiments from C2-WT and C2-H222P cells. Proteins extracted from C2-WT and C2-H222P cells were subjected to IP using antibodies against pERK1/2. Proteins in immunoprecipitates were separated by SDS-PAGE and IB using antibodies against pERK1/2 and cofilin-1. IgG was used as a negative control. Representative from three independent repeats. (C) Proteins extracted from C2-WT and C2-H222P cells were subjected to IP using antibodies specific to phospho Thr or phospho Ser. The immunoprecipitates were separated by SDS-PAGE and IB using antibody against cofilin-1. IgG was used as a negative control. Representative from three independent repeats. (D) Amino acid sequence of murine cofilin-1 with highlighted threonine 25 (red) and threonine 148 (blue). (E) C2-H222P cells transfected or not transfected (NT) with plasmids encoding cofilin-1, cofilin-1-T25A or cofilin-1-T148A were subjected to IP using antibodies against pERK1/2. Proteins in immunoprecipitates were separated by SDS-PAGE and IB using antibodies against cofilin-1 or pERK1/2. IgG was used as a negative control. Representative from three independent repeats. (F) Immunoblot illustrating the effect of transfection with different cofilin-1 constructs on the amount of G-actin and F-actin and the calculated F/G actin ratio. Data are represented as means±SEM (n=3; *P <0.05, **P <0.005) from three independent repeats. (G) Representative immunoblot of in vitro kinase assay using recombinant histidine-tagged wild-type cofilin-1 (His-cofilin-1-WT) and cofilin-1-T25A (His-cofilin-T25A), without or with the addition of recombinant pERK2. MBP was used as a positive control. [32P]-γ-ATP indicates recombinant protein phosphorylation. Corresponding Coomassie blue-stained gel is shown below immunoblot. Data in bar graph below are means±SEM (n=3; *P<0.05) from three independent repeats. (H) Representative immunoblot of in vitro kinase assay using recombinant His-cofilin-1, His-cofilin-1-T25A and His-cofilin-2 without or with the addition of recombinant pERK2. Phosphorylated (Thr25) cofilin-1 was detected using a specific antibody. Representative from three independent repeats. (I) Representative immunoblot of in vitro kinase assay using recombinant cofilin-1 without or with the addition of recombinant pERK2 at increasing doses. Phosphorylated (Thr25) cofilin-1 was detected using the specific antibody. Representative from three independent repeats.

Article Snippet: Radioactive in vitro kinase assay Cofilin-1, cofilin-1-T25A or dephosphorylated myelin basic protein (MBP) (Merck-Millipore) were incubated alone or in the presence of purified pERK2 in buffer comprising 50 mM HEPES (pH 7.5), 150 mM NaCl, 5 mM MgCl 2 , 0.5 mM TCEP and 0.5 mM ATP.

Techniques: Western Blot, SDS Page, Negative Control, Sequencing, Transfection, Construct, In Vitro, Kinase Assay, Recombinant, Positive Control, Staining

Journal: Human Molecular Genetics

Article Title: Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation

doi: 10.1093/hmg/ddy215

Figure Lengend Snippet: Phosphorylated (Thr25) cofilin-1 induces cardiac dysfunction. (A) Immunoblots showing cofilin-1, phosphorylated (Thr25) cofilin-1, cofilin-2 and gapdh in protein extracts from hearts of Lmna+/+ (Lmna WT) transduced with AAV vectors encoding cofilin-1; NT indicates not transduced. Representative from three independent repeats. (B) Immunoblot illustrating the effect of AAV expressing cofilin-1 construct on the amount of G-actin and F-actin and the calculated F/G actin ratio in hearts from mice; NT indicates not transduced. Data are represented as means±SEM (n=3; ***P<0.0005) from three independent repeats. (C) Immunoblots showing cofilin-1, phosphorylated (Thr25) cofilin-1, cofilin-2 and gapdh in protein extracts from hearts of LmnaH222P/H222P (Lmna H222P) transduced with AAV vectors encoding the indicated proteins; NT indicates not transduced. Representative from three independent repeats. (D) Immunoblot illustrating the effect of AAV expressing cofilin-1 constructs on the amount of G-actin and F-actin and the calculated F/G actin ratio in hearts from mice transduced with AAV vectors encoding the indicated proteins; NT indicates not transduced. Data are represented as means±SEM (n=3; *P<0.05, **P<0.005) from three independent repeats. (E) Graph showing left ventricular fraction shortening (FS) in 3-month-old (prior injection of AAV) and 6-month-old male transduced Lmna WT mice and Lmna H222P mice (see Table 2 for details). The central line represents the median, the box limits the interquartile range and the whiskers the minimum and maximum (*P<0.05, ***P<0.0005). (F) Expression of Col1a2, Myh7 and NppA mRNAs in hearts from 6-month-old male Lmna+/+ mice (Lmna WT) transduced with AAV expressing cofilin-1 or not transduced (NT). Data are represented as means±SEM (n=4; *P<0.05, **P <0.005, ***P <0.0005). (G) Expression of Col1a2, Myh7 and NppA mRNAs in hearts from 6-month-old male LmnaH222P/H222P mice (Lmna H222P) transduced with AAV expressing cofilin-1 or AAV expressing cofilin-1-T25A. Data from Lmna H222P mice not transduced (NT) are showed for comparison. Data are represented as means±SEM (n=4; *P<0.05, **P <0.005).

Article Snippet: Radioactive in vitro kinase assay Cofilin-1, cofilin-1-T25A or dephosphorylated myelin basic protein (MBP) (Merck-Millipore) were incubated alone or in the presence of purified pERK2 in buffer comprising 50 mM HEPES (pH 7.5), 150 mM NaCl, 5 mM MgCl 2 , 0.5 mM TCEP and 0.5 mM ATP.

Techniques: Western Blot, Transduction, Expressing, Construct, Injection

Journal: Human Molecular Genetics

Article Title: Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation

doi: 10.1093/hmg/ddy215

Figure Lengend Snippet: Phosphorylated (Thr25) cofilin-1 alters sarcomeric organization. (A) Micrographs showing α-actinin labeling of cross-sections of hearts from 6-month-old male Lmna+/+ (Lmna WT) mice and LmnaH222P/H222P (Lmna H222P) mice. Arrows indicate areas of disorganized sarcomeres (inset). (B) Immunoblots showing cofilin-1, pERK1/2, ERK1/2 and tubulin in protein extracts from CMs derived from patient-specific human iPSCs carrying the LMNA p.R190W mutation (LMNA R190W) and control (LMNA WT). Micrographs showing α-actinin labeling of CMs derived from patient-specific human iPSCs carrying LMNA p.R190W mutation (LMNA R190W) and control (LMNA WT). Dapi counter-staining (blue) of nuclei is also shown. Inset indicates altered myofibrillar organization. Arrows indicate Z-bands of sarcomeres. Means of organized and (black bar) and disorganized (while bar) sarcomeres are shown in each bar for each condition. (C) Electron micrographs showing disruption of sarcomeric organization in hearts from 6-month-old male LmnaH222P/H222P (Lmna H222P) mice compared with Lmna+/+ (Lmna WT) mice. Insets show a higher magnification. Arrows indicate disorganized sarcomeres. (D) Electron micrographs showing disorganized myofibrillar apparatus (a), areas without rods with normal sarcomere appearance (b) and sparse sarcomere structures (c) indicated by arrows in hearts from 6-month-old male LmnaH222P/H222P (Lmna H222P) mice. (E) Electron micrographs showing sarcomeric organization in hearts from 6-month-old Lmna+/+ (Lmna WT) mice transduced with AAV expressing cofilin-1 construct. NT indicates not transduced. Insets show a higher magnification. Arrows indicate disorganized sarcomeres. (F) Electron micrographs showing sarcomeric organization in hearts from 6-month-old LmnaH222P/H222P (Lmna H222P) mice transduced with AAV expressing cofilin-1 or cofilin-1-T25A constructs. Insets show a higher magnification. Arrows indicate disorganized sarcomeres. (G) Micrographs showing α-actinin labeling of CMs derived from human iPSCs transfected with plasmids encoding cofilin-1-T25A and cofilin-1-T25D. Insets highlight altered myofibrillar organization. Arrows indicate Z-bands of sarcomeres (inset). Dapi counter-staining (blue) of nuclei is also shown. (H) Micrographs showing α-actinin labeling of mouse HL-1 cardiac cells transfected with plasmids encoding cofilin-1-T25A and cofilin-1-T25D. Insets highlight altered myofibrillar organization. Arrows indicate Z-bands of sarcomeres (inset). Dapi counter-staining (blue) of nuclei is also shown. (I) Micrographs showing sarcomeric organization (MLC2vGFP) of embryonic mouse CMs transfected with plasmids encoding cofilin-1-T25A and cofilin-1-T25D. Insets highlight altered myofibrillar organization. Arrows indicate Z-bands of sarcomeres (inset).

Article Snippet: Radioactive in vitro kinase assay Cofilin-1, cofilin-1-T25A or dephosphorylated myelin basic protein (MBP) (Merck-Millipore) were incubated alone or in the presence of purified pERK2 in buffer comprising 50 mM HEPES (pH 7.5), 150 mM NaCl, 5 mM MgCl 2 , 0.5 mM TCEP and 0.5 mM ATP.

Techniques: Labeling, Western Blot, Derivative Assay, Mutagenesis, Staining, Transduction, Expressing, Construct, Transfection

Journal: Human Molecular Genetics

Article Title: Cofilin-1 phosphorylation catalyzed by ERK1/2 alters cardiac actin dynamics in dilated cardiomyopathy caused by lamin A/C gene mutation

doi: 10.1093/hmg/ddy215

Figure Lengend Snippet: A schematic representation of the mechanism of cofilin-1 phosphorylation by ERK1/2 in the heart from Lmna H222P mice with consequences on sarcomeric actin depolymerization. Blue circle indicates G-actin.

Article Snippet: Radioactive in vitro kinase assay Cofilin-1, cofilin-1-T25A or dephosphorylated myelin basic protein (MBP) (Merck-Millipore) were incubated alone or in the presence of purified pERK2 in buffer comprising 50 mM HEPES (pH 7.5), 150 mM NaCl, 5 mM MgCl 2 , 0.5 mM TCEP and 0.5 mM ATP.

Techniques: