Journal: IDCases

Article Title: An autopsy case of gas gangrene, massive intravascular hemolysis, and cytokine storm due to Clostridium perfringens type A infection

doi: 10.1016/j.idcr.2024.e02085

Figure Lengend Snippet: Patient with lethal Clostridium perfringens infection with massive intravascular hemolysis and gas gangrene. (a) Serum of the patient at presentation. (b) Abdominal computed tomography (CT) images obtained at presentation (left panel), 1.5 h (middle panel), and 2.5 h after death (right panel). The lesion in the right lobe of the liver, initially seen as a low-density area at presentation (left panel), was replaced by a gas-filled cavity 1.5 h later (middle panel). A postmortem CT scan revealed rapid and massive expansion of gas-filled cavities in the right and left lobes of the liver (right panel).

Article Snippet: Genomic PCRs targeting CPA, CPB, ETX, ITX, CPE, NetB, PFO, and ColA were performed using bacterial DNA of C. perfringens isolated from the patient’s blood and a commercially available type A strain of C. perfringens (JCM#1290; RIKEN BioResource Research Center, Tsukuba, Japan).

Techniques: Infection, Computed Tomography

Journal: IDCases

Article Title: An autopsy case of gas gangrene, massive intravascular hemolysis, and cytokine storm due to Clostridium perfringens type A infection

doi: 10.1016/j.idcr.2024.e02085

Figure Lengend Snippet: Host cytokine responses against Clostridium perfringens isolated from this case. (a) Toxin profiles of Clostridium perfringens isolated from the patient and a commercially available type A strain (JCM#1290). The leftmost lane represents the 100-bp DNA ladder. Agarose gel electrophoresis of polymerase chain reaction (PCR) products revealed that C. perfringens isolated from the patient expressed CPA, PFO, and ColA, but not CPE. CPA, C. perfringens α toxin; PFO, perfringolysin O; ColA, collagenase. (b) Profiles of serum cytokines and chemokines. Cytokine and chemokine arrays revealed heightened proinflammatory responses in the patient serum. CCL2; C-C chemokine ligand 2, CXCL8; C-X-C motif chemokine ligand 8, G-CSF; granulocyte-colony stimulating factor. (c) Toll-like receptors (TLRs) involved in the production of proinflammatory cytokines by C. perfringens . Splenocytes prepared from C57BL/6 mice or mice deficient in TLR2 and/or TLR4 were stimulated with heat-killed C. perfringens . IL-6 mRNA expression was expressed as the mean ± standard error of the mean. * * P < 0.01.

Article Snippet: Genomic PCRs targeting CPA, CPB, ETX, ITX, CPE, NetB, PFO, and ColA were performed using bacterial DNA of C. perfringens isolated from the patient’s blood and a commercially available type A strain of C. perfringens (JCM#1290; RIKEN BioResource Research Center, Tsukuba, Japan).

Techniques: Isolation, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Expressing

Journal: Biomedicines

Article Title: A Novel Homozygous Mutation in PMFBP1 Associated with Acephalic Spermatozoa Defects

doi: 10.3390/biomedicines13122882

Figure Lengend Snippet: The clinic and genetic data of the proband. ( a ) Images of embryos captured at 68 h post insemination using affected sperm or donor sperm, respectively. All embryos fertilized by mutant sperm were derived from 1PN zygotes, while embryos fertilized by donor sperm were derived from 2PN zygotes. ( b ) Validation of the PMFBP1 variant identified by WES through Sanger sequencing. The dashed box delineates the specific site of genetic variation, contrasting the mutant allele (T) identified in the patient with the conserved wild-type allele. (C) in the reference sequence. ( c ) Pedigree of the ASS family: Squares represent male family members, and circles represent female members. The mutation was validated by the parents of the proband. PMFBP1: polyamine modulated factor 1 binding protein 1; ASS: acephalic spermatozoa syndrome; M: mutation; WT: wild type.

Article Snippet: The mutant and wild-type (WT) PMFBP1 cDNAs were chemically synthesized by Sangon Biotech Company (Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China) and subsequently cloned into a pcDNA3.1+C-HA vector, which could express PMFBP1 WT or mutant +HA fusion protein with the HA tagged at the C-terminus end of PMFBP1 WT.

Techniques: Mutagenesis, Derivative Assay, Biomarker Discovery, Variant Assay, Sequencing, Binding Assay

Journal: Biomedicines

Article Title: A Novel Homozygous Mutation in PMFBP1 Associated with Acephalic Spermatozoa Defects

doi: 10.3390/biomedicines13122882

Figure Lengend Snippet: The analysis of the mutation in PMFBP1 . ( a ) The location of identified PMFBP1 variants and ours. Our variant is annotated in red lines in the genomic structure of PMFBP1 and the PMFBP1 protein. Full-length PMFBP1 is 1007 AA and the Smc domain is from 236 to 863 AA (green box). The p. Arg881Ter mutation causes the protein coding to terminate at the 881st arginine. ( b ) The PMFBP1 protein variant. The asterisk (*) denotes a premature termination codon at this amino acid position, resulting in a truncated protein product. ( c ) Alignment of multiple PFMBP1 amino acid sequences across species. ( d ) Structural changes in the PFMBP1 protein based on SWISS-MODEL. PMFBP1: polyamine modulated factor 1 binding protein 1; Smc: the functional structural maintenance of chromosome region; AA: amino acid.

Article Snippet: The mutant and wild-type (WT) PMFBP1 cDNAs were chemically synthesized by Sangon Biotech Company (Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China) and subsequently cloned into a pcDNA3.1+C-HA vector, which could express PMFBP1 WT or mutant +HA fusion protein with the HA tagged at the C-terminus end of PMFBP1 WT.

Techniques: Mutagenesis, Variant Assay, Binding Assay, Functional Assay

Journal: Biomedicines

Article Title: A Novel Homozygous Mutation in PMFBP1 Associated with Acephalic Spermatozoa Defects

doi: 10.3390/biomedicines13122882

Figure Lengend Snippet: Morphological ultrastructure defects in PMFBP1 mutant spermatozoa. ( a ) Light microscopy of Diff-Quik-stained normal sperm and the patient’s sperm. The blue arrows mark the head of the sperm, the red arrows highlight the tail of the sperm. Scale bar = 20 μm. ( b ) The acrosome and nucleus of the sperm are stained with TRITC-conjugated PNA (red) and DAPI (blue), respectively. ( c ) The TEM images of normal sperm and the patient’s sperm. The arrows indicate the disordered cervical mitochondrial sheath structures of the mutant sperm. PNA: Peanut Agglutinin; DAPI: 4′,6-diamidino-2-phenylindole; TEM: transmission electron microscopy.

Article Snippet: The mutant and wild-type (WT) PMFBP1 cDNAs were chemically synthesized by Sangon Biotech Company (Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China) and subsequently cloned into a pcDNA3.1+C-HA vector, which could express PMFBP1 WT or mutant +HA fusion protein with the HA tagged at the C-terminus end of PMFBP1 WT.

Techniques: Mutagenesis, Light Microscopy, Diff-Quik, Staining, Transmission Assay, Electron Microscopy

Journal: Biomedicines

Article Title: A Novel Homozygous Mutation in PMFBP1 Associated with Acephalic Spermatozoa Defects

doi: 10.3390/biomedicines13122882

Figure Lengend Snippet: Functional analysis of the PMFBP1 mutant. ( a ) The protein expression level decreased in the patient’s sperm after treatment. ( b ) Plasmid construction of the PMFBP1 WT and PMFBP1 mutant. ( c ) The stability of the protein increased after treatment with CHX (100 μmol/L) and the combination of CHX and MG132 (10 μmol/L). WT: Wild type; CHX: Cycloheximide.

Article Snippet: The mutant and wild-type (WT) PMFBP1 cDNAs were chemically synthesized by Sangon Biotech Company (Sangon Biotech (Shanghai) Co., Ltd., Shanghai, China) and subsequently cloned into a pcDNA3.1+C-HA vector, which could express PMFBP1 WT or mutant +HA fusion protein with the HA tagged at the C-terminus end of PMFBP1 WT.

Techniques: Functional Assay, Mutagenesis, Expressing, Plasmid Preparation