00fd896998d084096ad6de83638f7145 pmc_9570075.pdf f3c2bae3a7dd446423a0d081dbc1ab5ce8f7d24d pmc_9570075.pdf 1117dd485758f70b6f3f20835f63eaa4a4a992227822e4db47bdec8d4c8ba2cc pmc_9570075.pdf Title: Pathological Features in Paediatric Patients with TK2 Deficiency Subject: Thymidine kinase (TK2) deficiency causes mitochondrial DNA depletion syndrome. We aimed to report the clinical, biochemical, genetic, histopathological, and ultrastructural features of a cohort of paediatric patients with TK2 deficiency. Mitochondrial DNA was isolated from muscle biopsies to assess depletions and deletions. The TK2 genes were sequenced using Sanger sequencing from genomic DNA. All muscle biopsies presented ragged red fibres (RRFs), and the prevalence was greater in younger ages, along with an increase in succinate dehydrogenase (SDH) activity and cytochrome c oxidase (COX)-negative fibres. An endomysial inflammatory infiltrate was observed in younger patients and was accompanied by an overexpression of major histocompatibility complex type I (MHC I). The immunofluorescence study for complex I and IV showed a greater number of fibres than those that were visualized by COX staining. In the ultrastructural analysis, we found three major types of mitochondrial alterations, consisting of concentrically arranged lamellar cristae, electrodense granules, and intramitochondrial vacuoles. The pathological features in the muscle showed substantial differences in the youngest patients when compared with those that had a later onset of the disease. Additional ultrastructural features are described in the muscle biopsy, such as sarcomeric de-structuration in the youngest patients with a more severe phenotype. Keywords: TK2 deficiency; paediatric patients; ultrastructural studies; muscle biopsy; mitochondrial myopathies; ragged red fibres Author: Cristina Jou, Andres Nascimento, Anna Codina, Julio Montoya, Ester López-Gallardo, Sonia Emperador, Eduardo Ruiz-Pesini, Raquel Montero, Daniel Natera-de Benito, Carlos I. Ortez, Jesus Marquez, Maria V. Zelaya, Alfonso Gutierrez-Mata, Carmen Badosa, Laura Carrera-García, Jesica Expósito-Escudero, Monica Roldán, Yolanda Camara, Ramon Marti, Isidre Ferrer, Cecilia Jimenez-Mallebrera and Rafael Artuch Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Sep 22 15:39:09 2022 CEST ModDate: Thu Sep 22 15:44:03 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 7632801 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SXQPWE+VnURWPalladioL Type 1 Custom yes yes no 10 0 JFYKCX+URWPalladioL-Bold Type 1 Custom yes yes no 15 0 AFMNMZ+URWPalladioL-Roma Type 1 Custom yes yes no 20 0 XSPYYP+URWPalladioL-Ital Type 1 Custom yes yes no 24 0 KGRQPR+CMSY10 Type 1 Builtin yes yes no 28 0 PRJKGQ+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 50 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 53 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 81 0 MPAMEA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 92 0 MPAMEC+PalatinoLinotype TrueType WinAnsi yes yes no 95 0 MPAMFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 98 0 MPAMEA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 109 0 MPAMEC+PalatinoLinotype TrueType WinAnsi yes yes no 112 0 MPAMFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 115 0 MPAMEA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 130 0 MPAMEC+PalatinoLinotype TrueType WinAnsi yes yes no 133 0 MPAMFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 136 0 MPAMEA+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 151 0 MPAMEC+PalatinoLinotype TrueType WinAnsi yes yes no 154 0 MPAMFE+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 157 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-03-22 02:36:04 CET RepresentationInformation: pmc_9570075.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-10-27 02:17:00 CEST Size: 7632801 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 339 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Clinical, Biochemical, and Genetic Data of the Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Pathological Characteristics of Patients Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Immunofluorescence Studies for Mitochondrial Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Ultrastructural Examination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Children: Item: Title: Patients and Control Subjects Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Pathological Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Other Laboratory Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Info: Title: Pathological Features in Paediatric Patients with TK2 Deficiency Author: Cristina Jou, Andres Nascimento, Anna Codina, Julio Montoya, Ester López-Gallardo, Sonia Emperador, Eduardo Ruiz-Pesini, Raquel Montero, Daniel Natera-de Benito, Carlos I. Ortez, Jesus Marquez, Maria V. Zelaya, Alfonso Gutierrez-Mata, Carmen Badosa, Laura Carrera-García, Jesica Expósito-Escudero, Monica Roldán, Yolanda Camara, Ramon Marti, Isidre Ferrer, Cecilia Jimenez-Mallebrera and Rafael Artuch Subject: Thymidine kinase (TK2) deficiency causes mitochondrial DNA depletion syndrome. We aimed to report the clinical, biochemical, genetic, histopathological, and ultrastructural features of a cohort of paediatric patients with TK2 deficiency. Mitochondrial DNA was isolated from muscle biopsies to assess depletions and deletions. The TK2 genes were sequenced using Sanger sequencing from genomic DNA. All muscle biopsies presented ragged red fibres (RRFs), and the prevalence was greater in younger ages, along with an increase in succinate dehydrogenase (SDH) activity and cytochrome c oxidase (COX)-negative fibres. An endomysial inflammatory infiltrate was observed in younger patients and was accompanied by an overexpression of major histocompatibility complex type I (MHC I). The immunofluorescence study for complex I and IV showed a greater number of fibres than those that were visualized by COX staining. In the ultrastructural analysis, we found three major types of mitochondrial alterations, consisting of concentrically arranged lamellar cristae, electrodense granules, and intramitochondrial vacuoles. The pathological features in the muscle showed substantial differences in the youngest patients when compared with those that had a later onset of the disease. Additional ultrastructural features are described in the muscle biopsy, such as sarcomeric de-structuration in the youngest patients with a more severe phenotype. 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