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The Hunt Lab Guide to De Novo Peptide Sequence Analysis by Tandem Mass Spectrometry.

Donald Hunt has made seminal contributions to the fields of proteomics, immunology, epigenetics, and glycobiology. The foundation of every important work to come out of the Hunt Laboratory is de novo peptide sequencing. For decades, he taught hundreds of students, postdocs, engineers, and scientists to directly interpret mass spectral data. To honor his legacy and ensure that the art of de novo sequencing is not lost, we have adapted his teaching materials into "The Hunt Lab Guide to De Novo Peptide Sequence Analysis by Tandem Mass Spectrometry". In addition to the de novo sequencing tutorials, we present two freely available software tools that facilitate manual interpretation of mass spectra and validation of search results. The first, "Hunt Lab Peptide Fragment Calculator", calculates precursor and fragment mass-to-charge ratios for any peptide. The second program, "Predator Protein Fragment Calculator", was inspired in part by the fragment calculator developed in the Hunt Lab. Its capabilities are enhanced to facilitate interpretation of mass spectral data derived from intact proteins. We hope that the combination of these educational tools will continue to benefit students and researchers by empowering them to interpret data on their own.

Tandem Mass Spectrometry

Mass Spectrometry-Based Proteomics for the Masses: Peptide and Protein Identification in the Hunt Laboratory During the 2000's.

There has been a rapid increase in the number of individuals utilizing mass spectrometry-based proteomics to study complex biological systems and questions since the start of the 2000's. Building off the advancements in ionization and liquid chromatography scientists continued to push towards technology that would enable in-depth analysis of biological specimen. Donald F Hunt and the Hunt laboratory were major contributors to this effort with their work on improving upon existing Fourier Transform MS, development of electron transfer dissociation, and continued work on ion-ion reactions to improve intact protein analysis. Collaboration with other instrumentation laboratories and instrument companies led to the sharing of technology and eventual commercialization providing greater access. Additionally, the Hunt laboratory spread the gospel of MS-based proteomics through collaborations that lasted decades with other scientists who were experts in immunology, cellular signaling, epigenetics, and other fascinating fields. This article attempts to highlight the many contributions of Don and the Hunt laboratory to peptide and protein identification since the year 2000.

Humans

On the Hunt for the Histone Code.

Our genome is not made of naked DNA but a fiber (chromatin) composed of DNA and proteins packaged into our chromosomes. The basic building block of chromatin is the nucleosome, which has two copies of each of the proteins called histones (H2A, H2B, H3, and H4) wrapped by 146 base pairs of DNA. Regions of our genetic material are found between the more open (euchromatin) and more compact (heterochromatin) regions of the genome that can be variably accessible to the underlying genes. Furthermore, post-translational modifications (PTMs) on histones, such as on H3, are critical for regulating chromatin accessibility and gene expression. While site-specific antibodies were the tool of choice for histone PTM analysis in the early days (pre-2000s), enter Don Hunt changing the histone PTM field forever. Don's clever thinking brought new innovative mass spectrometry-based approaches to the epigenetics field. His lab's effort led to the discovery of many new histone modifications and methods to facilitate the detection and quantification of histone PTMs, which are still considered state of the art in the proteomics field today. Due to Don's pioneering work in this area, many labs have been able to jump into the epigenetics field and "Hunt" down their own histone targets. A walkthrough of those early histone years in the Hunt Lab is described by three of us who were fortunate enough to be at the right place, at the right time.

Animals

A Donald F. Hunt Story (John's Version).

A personal narrative of my time in the Hunt laboratory and beyond is provided. The impact of the Hunt laboratory on the analysis of peptides and proteins by tandem mass spectrometry is described in the context of the time.

History, 20th Century

Remittent painful ophthalmoplegia: the Tolosa-Hunt syndrome? A report of seven cases and review of the literature.

Seven patients with remittent painful ophthalmoplegia for which no specific local cause was found were seen during a period of five years. One had coincidental rheumatoid arthritis, and another had actinomycosis of the ipsilateral middle ear and contralateral parotid gland. The other five had no evidence of generalised autoimmune disease nor any other systemic disorder, two having separate episodes affecting each side. A history of relapsing and remitting painful ophthalmoplegia is suggestive of the Tolosa-Hunt syndrome, but it is rarely possible to confirm that the lesion in the cavernous sinus is the result of non-specific granulomatous infiltration so that the diagnosis remains one of exclusion. Carotid arteriography may show narrowing of the intracavernous part of the internal carotid artery. Orbital venography may also be helpful, particularly when the carotid arteriogram is normal. We believe that the Tolosa-Hunt syndrome is more common in England than is generally realised, but that its clinical features do not necessarily indicate a single pathological entity. Its recognition is important since the response to steroids, although not specific, is rapid in most patients, and the prognosis for complete recovery is relatively good.

Actinomycosis

Rifle wounds from high velocity, center-fire hunting ammunition.

Wounds inflicted by high velocity, center-fire rifles firing hunting ammunition are radically different from wounds caused by handguns or .22 rim-fire rifles. Injuries from pistol or .22 rim-fire bullets are confined to tissue and organs directly in the wound track. In contrast, high velocity rifle bullets can injure structures without actually contacting them. This is due to the temporary cavity produced by such missiles with the resultant shock waves having pressures of up to 200 atmospheres (20 MPa). Organs struck by such high velocity rifle bullets may undergo partial or complete disintegration. Hunting ammunition, as it passes through the body, tends to shed fragments of lead from its core, producing a characteristic snowstorm picture on X-ray. The maximum range at which powder tattooing of the skin occurs from center-fire rifles depends on the physical form of smokeless powder used as propellant. In a test with a 30-30 rifle, cartridges loaded with ball powder produced powder tattooing out to a range of 30 in. (76 cm), while similar cartridges, loaded with the traditional cylindrical powder, produced tattooing out to only 12 in. (30 cm) of range.

Firearms

The role of mental illness in the European witch hunts of the sixteenth and seventeenth centuries: an assessment.

Historians of psychiatry have propagated the view that the witch hunts of sixteenth and seventeenth-century Europe were primarily a persecution of the mentally ill and that demonological concepts of possession and witchcraft impeded psychiatric progress for centuries. The author reviews the evidence marshaled by these historians and examines additional historical material bearing on the psychopathological view. He concludes that the role of mental disorder in the witch hunts has been overinflated by authors with an interest in promulgating the medical model of abnormal behavior. Furthermore, the psychopathological paradigm is based on an outmoded philosophy of science, which results in historical distortion and paradoxes, and on restriction and selectivity in the choice of evidence.

Attitude to Health

[Rare care of Ramsay Hunt syndrome associated with herpes zoster of the glossopharyngeal nerve].

The authors describe a rare coexistence of Ramsay Hunt syndrome with glossopharyngeal zoster in a 67-year-old patient. A trial of systematization of the nomenclature of Ramsay Hunt syndrome is suggested, on the basis of certain anatomophysiological data. It is concluded that in zoster involvement of the sensory elements in the geniculate ganglion and in otic ganglion should be accompanied by taste sensitivity disturbances, but often the patients fail to notice these disturbances and routine taste testing shows also no such tase impairment. Routine use of electrogustometry is postulated since this makes possible diagnostic-prognostic assessment followed by selection of appropriate treatment.

Aged

Brain damage in National Hunt jockeys.

Five National Hunt jockeys have been found to have post-traumatic encephalopathy- three with epilepsy and two with significant intellectual and psychological deterioration. Closer supervision is needed.

Adult

Orbital venography in painful ophthalmoplegia (Tolosa-Hunt syndrome).

Twenty-six patients with painful ophthalmoplegia were classified as having the Tolosa-Hunt syndrome on the basis of their clinical findings and response to steroid therapy. All patients underwent orbital venography, a review of which forms the contents of this investigation. Orbital venography, a review of which forms the contents of this investigation. Orbital venograms were normal in 16 patients and abnormal in 10. The major abnormalities were (1) obstruction of the superior ophthalmic vein in its third segment without displacement; (2) collateral venous flow through small venous channels or collateral veins; and (3) poor opacification of the ipsilateral cavernous sinus.

Adolescent

[The Tolosa-Hunt syndrome (author's transl)].

This paper reports 2 cases of the Tolosa-Hunt Syndrome and reviews 70 other cases reported in the literature. Common findings of this syndrome are the recurring painful ophthalmoplegia affecting the 3rd, 4th and 6th cranial nerve, as well as the first branch of the 5th and the optic nerve. No ethiologic factor has so far been found, but is regarded as a non specific inflammation in the area of the sinus cavernosus and adjacent orbital bone. Residual defects in about one third of affected persons is the normal course of the disease. Early diagnosis and initiation of steroid therapy results in quick relief of pain and may influence prognosis.

Adolescent

The Ramsay Hunt syndrome: a dynamic demonstration of applied anatomy.

The clinical features of herpes zoster of the geniculate ganglion were reviewed and related to the functional anatomy of the seventh cranial nerve. An illustrative case highlights many of the features of the syndrome that was first described by Ramsay Hunt. The significance of facial palsy is discussed in the light of its distribution.

Aged

[Control of animal rabies by veterinary official and hunting measures (author's transl)].

The system of state control of rabies consists of clearing and eliminating the sources of infection, the simultaneous interruption of the chains of infection and preventing such chains developing. For this purpose different measures are instituted for rabies in domestic and wild animals. The weight of the attack lies in the control of rabies in the fox as the epidemiologically responsible carrier. Its reduction also corresponds to other gamekeeping requirements, the gassing of fox-earths being called upon when hunting measures fail. Vaccination of domestic animals, particularly the carnivores, presents an opportunity to break down epidemiological bridges for game and to reduce the danger to humans. Along with the improvement of vaccination of humans against rabies, this can contribute to a "dedramatisation" of rabies.

Animals

Hand-rearing of Cape hunting dog Lycaon pictus pups.

Problems encountered in the hand-rearing of Cape hunting dogs are discussed. It is felt that these could be minimised by avoiding stress, by rearing pups as close to natural circumstances as in possible, by applying sound husbandry techniques and by keeping the pups together.

Animal Husbandry

Reconstruction after total gastrectomy: construction of a Hunt-Lawrence pouch using Auto Suture staples.

A simple gastric reservoir using autosuture staples can be constructed rapidly after total gastrectomy. The importance of both a reservoir capacity and delayed evacuation was supported by the clinical course of most of the patients. All 18 patients had improved nutrition. Patients resected for cure tended to have greater weight gain and less abdominal discomfort. Occasionally patients who underwent only palliative resection were long-term survivors with a clinical course similar to that of patients resected for cure. On the basis of these results, it is unfair not to consider this type of reconstruction in patients who have undergone a total gastrectomy because it adds little to the operating time, yet it provides better palliation in terms of nutrition and abdominal comfort on both a short- and long-term basis.

Gastrectomy