Letter from 1475 contains proteins suggesting he suffered from hemolacria, respiratory problems.

The eponymous villain of Bram Stoker’s classic 1897 novel Dracula was partly inspired by a real historical person: Vlad III, a 15th century prince of Wallachia (now southern Romania), known by the moniker Vlad the Impaler because of his preferred method of execution: impaling his victims on spikes. Much of what we know about Vlad III comes from historical documents, but scientists have now applied cutting-edge proteomic analysis to three of the prince’s surviving letters, according to a recent paper published in the journal Analytical Chemistry. Among their findings: the Romanian prince was not a vampire, but he may have wept tears of blood, consistent with certain legends about Vlad III.

Vlad III was the second son of Vlad Dracul (“the Dragon”), who became the voivode of Wallachia in 1436. Vlad III was also known as Vlad Dracula (“son of the Dragon”), and it was this name that Stoker used for his fictional vampire—dracul means “the devil” in modern Romanian—along with a few historical details he was able to glean about Wallachia. This was a brutal, bloody period of political instability. Vlad spent several years as a prisoner of the Ottoman Empire, along with his younger brother Radu, and his father and older brother, Mircea, were murdered in 1447. Eventually, Vlad became voivode of Wallachia himself—three times, in fact, interrupted by periods of exile or captivity.

Vlad was constantly at war, and it was his brutal treatment of his enemies that led to his reputation as a monster, particularly in German-speaking territories, where books detailing his atrocities became bestsellers. These accounts described how Vlad executed men, women, and children taken prisoner from a Saxon village and impaled them. The more accurate, eye-witness-based accounts also included details about the churches Vlad’s army destroyed during plundering raids in Transylvania. Other stories (many likely exaggerated) claimed he burned the lazy and the poor, and had women impaled along with their nursing babies. A well-known woodcut shows Vlad dining while surrounded by impaled people on poles. He died in battle in January 1477, having killed an estimated 80,000 people in his lifetime.

We have some idea of what Vlad looked like, thanks to contemporary descriptions that admittedly are a bit biased by the prince’s brutal reputation. For instance, papal legate Nicholas of Modrussy described Vlad as “not very tall, but stocky and strong, with a cruel and terrible appearance, a long straight nose, distended nostrils, a thin and reddish face in which the large wide-open green eyes were framed by bushy black eyebrows, which made them appear threatening.” And there are several accounts, per the authors, of Vlad shedding tears of blood.
Ambras Castle’s portrait of Vlad III (c. 1560), reputedly a copy of an original made during his lifetime.
Enlarge / Ambras Castle’s portrait of Vlad III (c. 1560), reputedly a copy of an original made during his lifetime.
Public domain

Hoping to learn more about Vlad and the general environment in which he lived, the authors of the new study turned to three letters written by Vlad Dracula addressed to the rulers of the city of Sibiu. The first two were written in 1475, one of which includes Vlad’s personal signature; those letters have been stored in the Sibiu archives for more than 500 years and were never subjected to any kind of restoration efforts. The third letter was written in 1457 and was restored in Bucharest in the 20th century, although the authors state that the process was carried out in such a way as to minimize any biological or chemical contamination of the document.

Ancient biomolecules can tell us a lot about the distant past. For instance, bimolecular analysis has been used to shed light on painting materials, such as the beer byproducts used as canvas primers by painters during the Danish Golden Age; to learn more about ancient dietary practices; to explore whether ancient Egyptian followers of a deity called Bes used hallucinogens; and to analyze the microbes found on several of Leonardo da Vinci’s drawings.

Mass spectrometry-based proteomics is a relative newcomer to the field, capable of providing a thorough and very detailed characterization of any protein residues present in a given sample, as well as any accumulated damage. The technique is so sensitive that less sample material is needed compared to other methods. It’s also capable of characterizing all proteins present in a sample (regardless of the complexity of the mixture) rather than being narrowly targeted to predefined proteins. But collecting samples from such rare historical documents remains a challenge.

The authors, in this case, turned to a promising non-invasive sample collection technique called EVA (ethylene-vinyl-acetate). They used a plastic EVA film studded with strong cation and anion exchangers and certain hydrophobic resins, which extracted proteins and small molecules from the surfaces of the three letters. Those proteins and molecules were then analyzed via mass spectrometry. The team focused in particular on the oldest, most degraded proteins, reasoning that those were mostly likely to be from Vlad himself.

All told, the team identified 100 ancient human peptides—31 of which were deemed of particular interest—and an additional 2,000 peptides from bacteria, viruses, insects, fungi, and green plants. Those 31 human peptides were related to blood proteins or the respiratory system, as well as ciliopathy or retinal diseases, or inflammatory processes, per the authors. One of the letters from 1475 contained three peptides specifically associated with proteins of the eye’s retina and tears. The authors thus concluded that Vlad III may have suffered from a medical condition known as hemolacria, in which a person sheds tears of blood, as well as skin inflammation and respiratory illness. He may also have been exposed to plague-related bacteria or fruit flies and other pests, based on the non-human peptides analyzed.

There are caveats, of course, most notably the fact that the authors cannot entirely rule out the possibility that the human proteins could have come from other medieval people who may also have handled the documents. However, “It is also presumable that the most ancient proteins should be related to Prince Vlad the Impaler, who wrote and signed these letters,” the authors concluded.

Thanks to @boem for the link.

Wikipedia on Vlad