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Post by Admin on Mar 22, 2023 18:13:42 GMT
NEW YORK (AP) — Nearly 200 years after Ludwig van Beethoven’s death, researchers pulled DNA from strands of his hair, searching for clues about the health problems and hearing loss that plagued him. They weren’t able to crack the case of the German composer’s deafness or severe stomach ailments. But they did find a genetic risk for liver disease, plus a liver-damaging hepatitis B infection in the last months of his life. These factors, along with his chronic drinking, were probably enough to cause the liver failure that is widely believed to have killed him, according to a study published Wednesday in the journal Current Biology. This Sunday marks the 196th anniversary of Beethoven’s death in Vienna on March 26, 1827, at the age of 56. The composer himself wrote that he wanted doctors to study his health problems after he died. “With Beethoven in particular, it is the case that illnesses sometimes very much limited his creative work,” said study author Axel Schmidt, a geneticist at University Hospital Bonn in Germany. “And for physicians, it has always been a mystery what was really behind it.” Since his death, scientists have long tried to piece together Beethoven’s medical history and have offered a variety of possible explanations for his many maladies. Genomic analyses of hair from Ludwig van Beethoven Open Access Published:March 22, 2023DOI:https://doi.org/10.1016/j.cub.2023.02.041 www.cell.com/current-biology/fulltext/S0960-9822(23)00181-1Summary Ludwig van Beethoven (1770–1827) remains among the most influential and popular classical music composers. Health problems significantly impacted his career as a composer and pianist, including progressive hearing loss, recurring gastrointestinal complaints, and liver disease. In 1802, Beethoven requested that following his death, his disease be described and made public. Medical biographers have since proposed numerous hypotheses, including many substantially heritable conditions. Here we attempt a genomic analysis of Beethoven in order to elucidate potential underlying genetic and infectious causes of his illnesses. We incorporated improvements in ancient DNA methods into existing protocols for ancient hair samples, enabling the sequencing of high-coverage genomes from small quantities of historical hair. We analyzed eight independently sourced locks of hair attributed to Beethoven, five of which originated from a single European male. We deemed these matching samples to be almost certainly authentic and sequenced Beethoven’s genome to 24-fold genomic coverage. Although we could not identify a genetic explanation for Beethoven's hearing disorder or gastrointestinal problems, we found that Beethoven had a genetic predisposition for liver disease. Metagenomic analyses revealed furthermore that Beethoven had a hepatitis B infection during at least the months prior to his death. Together with the genetic predisposition and his broadly accepted alcohol consumption, these present plausible explanations for Beethoven’s severe liver disease, which culminated in his death. Unexpectedly, an analysis of Y chromosomes sequenced from five living members of the Van Beethoven patrilineage revealed the occurrence of an extra-pair paternity event in Ludwig van Beethoven’s patrilineal ancestry.
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Post by Admin on Mar 23, 2023 19:08:53 GMT
Introduction On March 27, 1827, the day after Beethoven’s death, two of his associates discovered several documents stored within a hidden compartment in his writing desk, including an unusual document written in 1802 and addressed to his brothers, which is now known as the Heiligenstadt Testament. In it, Beethoven confessed that he had been “hopelessly afflicted” with a progressive hearing loss. Stating that only virtue and his art held him back from committing suicide, he explained that he could not leave the world “before I had produced all the works that I felt the urge to compose.” Beethoven then requested that following his death, his disease be described by his favorite physician, Dr. Johann Adam Schmidt (1759–1809), and made public.1
Although Beethoven outlived Dr. Schmidt by 18 years, medical biographers have since attempted to determine the most likely causes of Beethoven’s various health complaints. Such research has relied principally on documentary sources, including Beethoven’s letters, diaries, and conversation books, and accounts from Beethoven’s contemporaries including physicians’ notes, an autopsy report, and descriptions of skeletal material following exhumations in 1863 and 1888. In addition, analyses of tissue sources claimed to originate from Beethoven have been performed, including toxicological analyses of hairs of unknown authenticity2,3,4 and paleopathological and toxicological examinations of skull fragments,5 at least two of which are inauthentic.6 These sources attest to a number of health complaints varying in severity and impact on Beethoven’s life and career. Foremost among these were a bilateral, late-onset, progressive, and predominantly sensorineural form of hearing loss, as well as chronic gastrointestinal problems and, toward the end of Beethoven’s life, liver disease. Beethoven’s hearing loss began in his mid- to late 20s, characterized initially by tinnitus, loudness-recruitment, and the loss of high-tone frequencies, and would end his career as a performing artist by his mid-40s.7,8 From at least the age of 22, Beethoven suffered from debilitating abdominal complaints that continued throughout his adult life, characterized primarily by abdominal pains (“Kolik”) and attacks and remissions of often prolonged bouts of diarrhea. In the summer of 1821, Beethoven began to exhibit symptoms of liver disease when the first of at least two attacks of jaundice occurred, culminating in his death, considered most likely due to cirrhosis,7,8 on March 26, 1827. Several lines of evidence indicative of the regular consumption of moderate to large quantities of alcohol9 have led some medical biographers to conclude that Beethoven was alcohol dependent,8,10 which is a known risk factor for liver cirrhosis.11 While several of Beethoven’s contemporaries insisted that Beethoven usually consumed alcohol in moderation,12,13,14 one close friend is alleged to have stated that in ca. 1825–1826, Beethoven had been consuming at least a liter (“Mass”) of wine with lunch every day.9 Although little is known with certainty about the medical history of Beethoven’s immediate family, a family history of alcohol dependence and liver disease has been noted.7,8 In addition to the three areas of illness mentioned above, Beethoven also showed other symptoms during his life, somatic and possibly also psychological.7,8,9 In clarifying possible genetic causes of Beethoven’s illnesses, we limit our investigation to the three somatic disease areas that dominate the medical biographical literature because they represent Beethoven’s main health restrictions and are widely documented by Beethoven’s own reports, as well as reports from Beethoven’s contemporaries and physicians. We sought to sequence Beethoven’s whole genome to high coverage from authenticated strands of hair in order to improve our understanding of his health. On the basis of genetic data and supporting provenance information, we assessed the authenticity of eight locks of hair claimed to originate from Beethoven. We required authentic samples to derive from a single male individual and to exhibit DNA damage patterns consistent with the reported antiquity of the samples. We sequenced a 24-fold coverage genome from the best-preserved sample among five matching samples using a highly sensitive protocol for historical hair. We then performed ancestry analyses on the expectation that this individual’s ancestry would be consistent with Beethoven’s documented genealogy. As part of our ancestry analyses, we introduce a novel geo-genetic triangulation (GGT) technique using long identity-by-descent (IBD) segments shared with individuals in FamilyTreeDNA’s genealogically explicit consumer database to determine the likely locations of Beethoven’s ancestors. In addition, we compared this genome to two groups of genealogically documented living relatives. We extensively analyzed Beethoven’s genome for genetic causes of and risk for somatic disease, in addition to metagenomic screening for evidence of infections, followed by targeted DNA capture.
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Post by Admin on Mar 24, 2023 19:02:18 GMT
Results Authentication of hair samples Strands of hair from eight locks attributed to Ludwig van Beethoven were acquired from public and private collections for analysis, which were determined to have had independent provenances (Figure 1; STAR Methods). We refer to these as the Müller, Bermann, Halm-Thayer, Moscheles, Stumpff, Cramolini-Brown, Hiller, and Kessler Locks (Methods S1A–S1H). These locks can conservatively be estimated to fall within a transect of Beethoven’s life between ca. November of 1821 and his death in March of 1827, with the provenance histories for two of these locks, the Stumpff and Halm-Thayer Locks, bearing intact chains of custody (Figure 1). We expected authentic and independently sourced locks of Beethoven’s hair to derive from a single male with predominantly Central European ancestry. We further expected the presence of terminal C-T deamination caused by DNA degradation over time, consistent with their provenances in the early 19th Century. Figure 1 Provenance summaries and authenticities of hair locks Timelines, sex determination, mitochondrial and Y chromosome haplogroup determinations, and authenticity assessments for eight tested locks of hair claimed to have originated from Beethoven. The Kessler lock has a date of acquisition 36 to 61 years after Beethoven’s death, during one of his exhumations in 1863 or 1888. See also Tables S1 and S2 and Figure 5. We performed shallow shotgun sequencing to permit assessments of DNA preservation and authenticity (Figure 1; Data S1A). Five samples, the Müller, Bermann, Halm-Thayer, Moscheles, and Stumpff Locks, shared identical mitochondrial genomes of haplogroup H1b1+16,362C with a private mutation at C16,176T, and had male XY karyotypes (Figure 1; Table S2; STAR Methods). Relatedness testing of autosomal and X chromosome DNA demonstrated that these five matching samples originated either from a single individual or monozygotic twins (Figure 2; Table S5; STAR Methods). All matching samples had DNA damage patterns consistent with their provenances in the early 19th Century (Data S1A).
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Post by Admin on Mar 25, 2023 18:40:25 GMT
Figure 2 Relatedness testing of eight locks of hair attributed to Beethoven Relatedness testing of eight locks of hair attributed to Beethoven, relative to an external reference panel of 41 medieval Bavarians. The proportions of non-matching alleles (P0) per pair and estimated degrees of relatedness are calculated from pseudo-haploid genotype calls using READ. The numbers of SNVs in each comparison are denoted by n. ∗Known medieval Bavarian sibling pair (STR355c/STR491). See also Table S5. In light of their provenance histories, we considered these genetic findings to be compelling evidence for the identity of these five independent locks of hair and proceeded under the working hypothesis that they are authentic. We determined that the Stumpff Lock (Figure 3) was marginally the best preserved of the five matching samples (Methods S1I) and sequenced a nuclear genome to an average of 24-fold coverage, incorporating laboratory and bioinformatics protocols optimized for the ultra-short DNA fragments characteristic of historical hair samples (mean fragment length 29.62 bp)15,16,17,18,19,20,21 (STAR Methods). We restricted further analyses to the 1.64 Gb of the genome to which short reads could be confidently mapped (“accessible genome”; STAR Methods). Figure 3 The Stumpff Lock The Stumpff Lock from which Beethoven’s high-coverage genome was sequenced. The lock is affixed to a letter from Johann Andreas Stumpff to Patrick Stirling, dated May 7, 1827. Stumpff’s poem reads, “The head⦻, these hair’s have grac’d lies low; But what it wrought — will ever grow. ⦻of Lud. v. Beethoven.” Photographed in July of 2018 by American Beethoven Society member Kevin Brown.
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Post by Admin on Mar 26, 2023 19:11:16 GMT
Figure 4 Geo-genetic triangulation of Beethoven’s probable ancestor locations Geo-genetic triangulation, showing probable locations of Beethoven’s autosomal ancestors. Regional intensity per hexagon represents number of shared ancestors between Beethoven and modern FTDNA customers, with the following criteria: (1) IBD segments (n = 665) must be shared (triangulated) between at least three individuals to be genetically validated and (2) ancestor locations (n = 89) of matching individuals must occur within the same hexagon to be geographically validated. Areas of Western and Central Europe that were considered but lack any probable ancestors are shown in gray. See also Methods S1L–S1O. Of the non-matching hair samples tested, our sequence data show that the highly publicized Hiller Lock originated from a woman with close autosomal affinity in PCA space to present day North African, Middle Eastern, and Jewish populations24 (Methods S1K). Its mitochondrial haplogroup, K1a1b1a (Table S2), is highly prevalent among Ashkenazi Jews.25 Toxicological analyses of hairs extracted from this lock have been used to argue that Beethoven’s health problems were caused or compounded by plumbism, and to refute suggestions that he was administered opiates during the course of his final illness and mercury for a hypothesized infection with syphilis.2,3,4,26,27 We now conclude that these findings do not apply to Beethoven. We additionally demonstrate that patterns of longitudinally distributed lead isotope concentrations believed to have been shared between hair strands from the Hiller, Halm-Epstein, and Erdödy Locks3 do not constitute proof of their authenticity, as the Hiller Lock is inauthentic. We found that the Cramolini-Brown Lock originated from a male of European autosomal ancestry (Figure 1; Methods S1J), belonging to the mitochondrial haplogroup H79 and the Y chromosome haplogroup R1a-Z283 (Figures 1 and S4). As this sample differs from our five matching samples and we could not confirm its provenance prior to 2012, we conclude that it is almost certainly inauthentic. Both the Hiller and Cramolini-Brown Locks exhibited levels of DNA damage similar to the five matching Beethoven samples (Data S1A).28,29 The Kessler Lock lacked sufficient DNA preservation for sex chromosomal karyotyping or ancestry determination, mitochondrial contamination estimation, or mitochondrial haplogroup assignment (Data S1A). We were therefore unable to assess its authenticity. On the basis of these genetic data, and in light of their known provenance histories, we conclude that the Müller, Bermann, Halm-Thayer, Moscheles, and Stumpff Locks almost certainly authentically derive from Beethoven, the Cramolini-Brown Lock is almost certainly inauthentic, the Hiller Lock is definitely inauthentic, and the authenticity of the Kessler Lock could not be determined. In order to support the authenticity of the matching samples further, we compared the Y chromosome from Beethoven’s high-coverage genome against high-coverage Y chromosomes sequenced from five living men belonging to the Van Beethoven patrilineage. These individuals were identified through analyses of genealogical records, which document Aert van Beethoven (1535–1609) as a patrilineal ancestor shared by Ludwig van Beethoven and our research participants (Methods S1P).23 Consistent with genealogical records, these five individuals share nearly identical Y chromosomes falling within the R-FT446200 haplogroup within R1b, with an average of 4.8 [3–7] private mutations having arisen along each lineage (Figure 5; STAR Methods). The pedigree reconstructed on the basis of these private mutations reproduced the documented pedigree among the participants (Figure 5). These Y chromosomes did not, however, match the Y chromosomes from either the five matching Beethoven hair samples within I1a-Z139 or the Cramolini-Brown Lock within R1a-Z283 (Figure 5).
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