A re-assessment of Dr Robert Knox and his contribution to early evolution science.

Dr Robert Knox was publicly scorned and disgraced for his unwitting involvement in the Burke and Hare serial murders in 1828. Far less appreciated is his brilliance as an anatomist and he espoused the European movement in Transcendental Anatomy, which aimed to uncover the laws governing what we now know as evolution and the origin of species. Knox fully embraced Transcendental Anatomy during a sojourn in Paris and taught it on his return to Edinburgh, where there was a critical mass of like-minded Transcendental Anatomists. Charles Darwin spent 1825-1827 as a medical student in Edinburgh when Transcendental Anatomy was at its peak amongst the city's anatomists, and evolution - then known as transmutation - was a source of great interest and controversy. Knox intended to research Transcendental Anatomy, but this was thwarted by conflicting demands on his time in the second half of the 1820s decade and the Burke and Hare tragedy. He did, however, go on to champion Transcendental Anatomy and write extensively on it.

Richard Muir: Edinburgh-based pioneer biomedical scientist and medical artist.

Richard Muir (1862-1931) began his career as a 'lab boy' in the Pathology Department of the University of Edinburgh in 1876 at the age of 13. This was a newly created category of worker that eventually became today's biomedical scientist Muir rapidly gained expertise in pathological and bacteriological techniques including staining and microscopy. Exceptionally, for someone non-medical and non-university-educated individual, he was elected a member of the Pathological Society of Great Britain and appointed Demonstrator in Pathology in the University of Edinburgh Pathology Department. He authored papers on staining techniques for bacteria and on the pathology of syphilis of the ear and became a recognised diagnostic histopathologist, despite having no medical qualifications. He especially excelled as an artist, depicting the microscopic world of pathology and microbiology and produced diagrams for hundreds of publications including his own book and also large wall hangings of the microscopic world for teaching purposes. This paper describes the unique contribution of Richard Muir to pathology in Edinburgh and beyond in the early 20th century.

The recognition of lung disease in coal workers: The role of Gough-Wentworth whole lung sections.

From the identification of a specific lung disease caused by coal dust exposure in miners in 1831 until the demonstration of the association of that exposure to risk of emphysema in 1984, there was continuous argument about the harmfulness of coal dust. Ill health in miners was attributed variously to tuberculosis, quartz exposure or cigarette smoking. An acceptance that coal dust was harmful only started with investigative radiology and pathology in the 1920s, and physiology in the 1950s. Most of the early investigations were in South Wales, the centre of the most important coal field in Great Britain. Among the investigators was Professor Jethro Gough who, with his technician James Wentworth, introduced a technique for making thick sections of whole, inflated lungs on paper backing. Here, we describe this method and its central role in understanding the relationships between coal dust exposure, pneumoconiosis, emphysema and lung dysfunction in miners.

Dr Robert Knox and his book on fishing in Scotland: A window into his mind.

Robert Knox was publicly vilified for his suspected complicity in the 16 murders committed by Burke and Hare, although he had no involvement in them. Along with several books on anatomy Knox also wrote a book on angling in Lowland Scotland. In 'Fish and Fishing in the Lone Glens of Scotland' Knox's deep love for nature and for fishing emerges. Most interesting however is that although generally focussed on fish and fishing, the book abounds with asides on Knox's other preoccupations and passions. These provide rare insights into the character of the great anatomist, whose personality has otherwise retained its opacity over the years. In the book, Knox writes in passing and in a relatively unguarded fashion, about such topics as transcendental anatomy, Scottish Independence, empiricism, race and Edinburgh medical figures. In so-doing, we contend that he affords the reader some insight into the mind of the real Dr Robert Knox.

John Goodsir and local opposition to Rudolf Virchow's election to Fellowship of the Royal Society of Edinburgh in 1868.

In 1869 Rudolf Virchow, the distinguished Prussian pathologist who pioneered the modern concept of cellular pathology, was offered an honorary Fellowship of The Royal Society of Edinburgh (RSE). However, the Rev. Joseph T Goodsir, Fellow of the Royal Society of Edinburgh (FRSE), the brother of Professor John Goodsir FRSE, the famed Edinburgh anatomist who had died two years previously, mounted a campaign to stop the award. As part of this he published a pamphlet entitled Grounds of Objection to the Admission of Professor Virchow as an Honorary Fellow of the Royal Society of Edinburgh. The disagreement centred on John Goodsir's pioneering research and writings on cell theory. These had in fact been recognised by Virchow, who dedicated the English language edition of his most famous publication Cellular Pathology to John Goodsir. Joseph Goodsir was not, however, satisfied by this and the basis of his objection was that Virchow had plagiarised from his brother. We describe the background and outcome of this dispute.

John Goodsir: discovering Sarcina ventriculi and diagnosing Darwin's dyspepsia.

In 1842, when John Goodsir was Conservator to the Museum of the RCSEd, he saw a 19-year-old male patient who vomited a large volume of acidic, fermented-smelling, watery fluid every morning. Under his microscope, Goodsir found the vomitus to be populated with a micro-organism he named Sarcina ventriculi, which he considered to be causative. In so-doing, Goodsir became one of the first people to link a specific micro-organism with a disease. Goodsir recommended small doses of creosote as an antiseptic and claimed that the boy was eventually cured of the vomiting condition. In August of 1863 Charles Darwin was hugely celebrated by the scientific community and the public, but he had suffered from severe stomach problems all his adult life and at this point, he was vomiting daily. He read Goodsir's paper and contacted him and asked if he could send some vomitus samples to Edinburgh in the hope that Goodsir might find Sarcina in it and solve the mystery of his debilitating stomach symptoms and perhaps cure them with creosote. Goodsir examined samples in his microscope, but failed to find Sarcina. Sadly, Darwin went on to suffer constantly from severe stomach problems, recently attributed to lactose intolerance, until he died in 1882, some 20 years later.

Black lungs in the general population: a new look at an old dispute.

Almost from the time that autopsies were first routinely carried out, darkening of lungs with increasing age was described. Different explanations for the origin of the accumulating black pigment arose and by the early nineteenth century three hypotheses had emerged: 1) soot inhaled into the lungs from the air; 2) carbon accumulating in the lungs from abnormal pulmonary carbon dioxide metabolism; and, 3) pigment derived from the blood. In 1813 the English physician and chemist George Pearson published a paper in which he described the recovery of the black pigment from lungs and its chemical analysis. Pearson declared the black pigment to be airborne carbon/soot from the burning of coal and wood. He described these particles depositing in 'black spots' in the terminal airways and accumulating in the peribronchial lymph nodes, forming 'black glands'. Despite Pearson's prescient account, debate continued and the true explanation, given in that paper, was not fully accepted until the late nineteenth century.

Induction of protein citrullination and auto-antibodies production in murine exposed to nickel nanomaterials.

Citrullination, or the post-translational deimination of polypeptide-bound arginine, is involved in several pathological processes in the body, including autoimmunity and tumorigenesis. Recent studies have shown that nanomaterials can trigger protein citrullination, which might constitute a common pathogenic link to disease development. Here we demonstrated auto-antibody production in serum of nanomaterials-treated mice. Citrullination-associated phenomena and PAD levels were found to be elevated in nanomaterials -treated cell lines as well as in the spleen, kidneys and lymph nodes of mice, suggesting a systemic response to nanomaterials injection, and validated in human pleural and pericardial malignant mesothelioma (MM) samples. The observed systemic responses in mice exposed to nanomaterials support the evidence linking exposure to environmental factors with the development of autoimmunity responses and reinforces the need for comprehensive safety screening of nanomaterials. Furthermore, these nanomaterials induce pathological processes that mimic those observed in Pleural MM, and therefore require further investigations into their carcinogenicity.

Long-Fiber Carbon Nanotubes Replicate Asbestos-Induced Mesothelioma with Disruption of the Tumor Suppressor Gene Cdkn2a (Ink4a/Arf).

Mesothelioma is a fatal tumor of the pleura and is strongly associated with asbestos exposure. The molecular mechanisms underlying the long latency period of mesothelioma and driving carcinogenesis are unknown. Moreover, late diagnosis means that mesothelioma research is commonly focused on end-stage disease. Although disruption of the CDKN2A (INK4A/ARF) locus has been reported in end-stage disease, information is lacking on the status of this key tumor suppressor gene in pleural lesions preceding mesothelioma. Manufactured carbon nanotubes (CNTs) are similar to asbestos in terms of their fibrous shape and biopersistent properties and thus may pose an asbestos-like inhalation hazard. Here we show that instillation of either long CNTs or long asbestos fibers into the pleural cavity of mice induces mesothelioma that exhibits common key pro-oncogenic molecular events throughout the latency period of disease progression. Sustained activation of pro-oncogenic signaling pathways, increased proliferation, and oxidative DNA damage form a common molecular signature of long-CNT- and long-asbestos-fiber-induced pathology. We show that hypermethylation of p16/Ink4a and p19/Arf in CNT- and asbestos-induced inflammatory lesions precedes mesothelioma; this results in silencing of Cdkn2a (Ink4a/Arf) and loss of p16 and p19 protein, consistent with epigenetic alterations playing a gatekeeper role in cancer. In end-stage mesothelioma, silencing of p16/Ink4a is sustained and deletion of p19/Arf is detected, recapitulating human disease. This study addresses the long-standing question of which early molecular changes drive carcinogenesis during the long latency period of mesothelioma development and shows that CNT and asbestos pose a similar health hazard.

Assessment of the potential respiratory hazard of volcanic ash from future Icelandic eruptions: a study of archived basaltic to rhyolitic ash samples.

BACKGROUND: The eruptions of Eyjafjallajökull (2010) and Grímsvötn (2011), Iceland, triggered immediate, international consideration of the respiratory health hazard of inhaling volcanic ash, and prompted the need to estimate the potential hazard posed by future eruptions of Iceland's volcanoes to Icelandic and Northern European populations. METHODS: A physicochemical characterization and toxicological assessment was conducted on a suite of archived ash samples spanning the spectrum of past eruptions (basaltic to rhyolitic magmatic composition) of Icelandic volcanoes following a protocol specifically designed by the International Volcanic Health Hazard Network. RESULTS: Icelandic ash can be of a respirable size (up to 11.3 vol.% < 4 µm), but the samples did not display physicochemical characteristics of pathogenic particulate in terms of composition or morphology. Ash particles were generally angular, being composed of fragmented glass and crystals. Few fiber-like particles were observed, but those present comprised glass or sodium oxides, and are not related to pathogenic natural fibers, like asbestos or fibrous zeolites, thereby limiting concern of associated respiratory diseases. None of the samples contained cristobalite or tridymite, and only one sample contained quartz, minerals of interest due to the potential to cause silicosis. Sample surface areas are low, ranging from 0.4 to 1.6 m2 g-1, which aligns with analyses on ash from other eruptions worldwide. All samples generated a low level of hydroxyl radicals (HO•), a measure of surface reactivity, through the iron-catalyzed Fenton reaction compared to concurrently analyzed comparative samples. However, radical generation increased after 'refreshing' sample surfaces, indicating that newly erupted samples may display higher reactivity. A composition-dependent range of available surface iron was measured after a 7-day incubation, from 22.5 to 315.7 µmol m-2, with mafic samples releasing more iron than silicic samples. All samples were non-reactive in a test of red blood cell-membrane damage. CONCLUSIONS: The primary particle-specific concern is the potential for future eruptions of Iceland's volcanoes to generate fine, respirable material and, thus, to increase ambient PM concentrations. This particularly applies to highly explosive silicic eruptions, but can also hold true for explosive basaltic eruptions or discrete events associated with basaltic fissure eruptions.

Inhaled Nanoparticles Accumulate at Sites of Vascular Disease.

The development of engineered nanomaterials is growing exponentially, despite concerns over their potential similarities to environmental nanoparticles that are associated with significant cardiorespiratory morbidity and mortality. The mechanisms through which inhalation of nanoparticles could trigger acute cardiovascular events are emerging, but a fundamental unanswered question remains: Do inhaled nanoparticles translocate from the lung in man and directly contribute to the pathogenesis of cardiovascular disease? In complementary clinical and experimental studies, we used gold nanoparticles to evaluate particle translocation, permitting detection by high-resolution inductively coupled mass spectrometry and Raman microscopy. Healthy volunteers were exposed to nanoparticles by acute inhalation, followed by repeated sampling of blood and urine. Gold was detected in the blood and urine within 15 min to 24 h after exposure, and was still present 3 months after exposure. Levels were greater following inhalation of 5 nm (primary diameter) particles compared to 30 nm particles. Studies in mice demonstrated the accumulation in the blood and liver following pulmonary exposure to a broader size range of gold nanoparticles (2-200 nm primary diameter), with translocation markedly greater for particles <10 nm diameter. Gold nanoparticles preferentially accumulated in inflammation-rich vascular lesions of fat-fed apolipoproteinE-deficient mice. Furthermore, following inhalation, gold particles could be detected in surgical specimens of carotid artery disease from patients at risk of stroke. Translocation of inhaled nanoparticles into the systemic circulation and accumulation at sites of vascular inflammation provides a direct mechanism that can explain the link between environmental nanoparticles and cardiovascular disease and has major implications for risk management in the use of engineered nanomaterials.

Factors influencing withdrawal from dialysis: a national registry study.

BACKGROUND: Dialysis withdrawal is the third most common cause of death in patients receiving dialysis for established renal failure (ERF) in Scotland. We describe incidence, risk factors and themes influencing decision-making in a national renal registry. METHODS: Details of deaths in those receiving renal replacement therapy (RRT) for ERF in Scotland are reported to the Scottish Renal Registry via a unique mortality report. We extracted patient demographics and comorbidity, cause and location of death, duration of RRT and pertinent free text comments from 1 January 2008 to 31 December 2014. Withdrawal incidence was calculated and logistic regression used to identify significantly influential variables. Themes emerging from clinician comments were tabulated for descriptive purposes. RESULTS: There were 2596 deaths; median age at death was 68 [interquartile range (IQR) 58, 76] years, 41.5% were female. Median duration on RRT was 1110 (IQR 417, 2151) days. Dialysis withdrawal was the primary cause of death in 497 (19.1%) patients and withdrawal contributed to death in a further 442 cases (17.0%). The incidence was 41 episodes per 1000 patient-years. Regression analysis revealed increasing age, female sex and prior cerebrovascular disease were associated with dialysis withdrawal as a primary cause of death. Conversely, interstitial renal disease, angiographically proven ischaemic heart disease, valvular heart disease and malignancy were negatively associated. Analysis of free text comments revealed common themes, portraying an image of physical and psychological decline accelerated by acute illnesses. CONCLUSIONS: Death following dialysis withdrawal is common. Factors important to physical independence-prior cerebrovascular disease and increasing age-are associated with withdrawal. When combined with clinician comments this study provides an insight into the clinical decline affecting patients and the complexity of this decision. Early recognition of those likely to withdraw may improve end of life care.

SERS as a tool for in vitro toxicology.

Measuring markers of stress such as pH and redox potential are important when studying toxicology in in vitro models because they are markers of oxidative stress, apoptosis and viability. While surface enhanced Raman spectroscopy is ideally suited to the measurement of redox potential and pH in live cells, the time-intensive nature and perceived difficulty in signal analysis and interpretation can be a barrier to its broad uptake by the biological community. In this paper we detail the development of signal processing and analysis algorithms that allow SERS spectra to be automatically processed so that the output of the processing is a pH or redox potential value. By automating signal processing we were able to carry out a comparative evaluation of the toxicology of silver and zinc oxide nanoparticles and correlate our findings with qPCR analysis. The combination of these two analytical techniques sheds light on the differences in toxicology between these two materials from the perspective of oxidative stress.

Platelet activation independent of pulmonary inflammation contributes to diesel exhaust particulate-induced promotion of arterial thrombosis.

BACKGROUND: Accelerated thrombus formation induced by exposure to combustion-derived air pollution has been linked to alterations in endogenous fibrinolysis and platelet activation in response to pulmonary and systemic inflammation. We hypothesised that mechanisms independent of inflammation contribute to accelerated thrombus formation following exposure to diesel exhaust particles (DEP). METHODS: Thrombosis in rats was assessed 2, 6 and 24 h after administration of DEP, carbon black (CB; control carbon nanoparticle), DQ12 quartz microparticles (to induce pulmonary inflammation) or saline (vehicle) by either intra-tracheal instillation (0.5 mg, except Quartz; 0.125 mg) or intravenous injection (0.5 mg/kg). Thrombogenicity was assessed by carotid artery occlusion, fibrinolytic variables and platelet-monocyte aggregates. Measures of inflammation were determined in plasma and bronchoalveolar lavage fluid. Tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI)-1 were measured following direct in vitro exposure of human umbilical vein endothelial cells (HUVECs) to DEP (10-150 µg/mL). RESULTS: Instillation of DEP reduced the time to thrombotic occlusion in vivo, coinciding with the peak of DEP-induced pulmonary inflammation (6 h). CB and DQ12 produced greater inflammation than DEP but did not alter time to thrombotic occlusion. Intravenous DEP produced an earlier (2 h) acceleration of thrombosis (as did CB) without pulmonary or systemic inflammation. DEP inhibited t-PA and PAI-1 release from HUVECs, and reduced the t-PA/PAI-1 ratio in vivo; similar effects in vivo were seen with CB and DQ12. DEP, but not CB or DQ12, increased platelet-monocyte aggregates. CONCLUSION: DEP accelerates arterial thrombus formation through increased platelet activation. This effect is dissociated from pulmonary and systemic inflammation and from impaired fibrinolytic function.

Surface charge determines the lung inflammogenicity: A study with polystyrene nanoparticles.

Surface functionalization is a routine process to improve the behavior of nanoparticles (NPs), but the induced surface properties, such as surface charge, can produce differential toxicity profiles. Here, we synthesized a library of covalently functionalized fluorescent polymeric NPs (F-PLNPs) to evaluate the role of surface charge on the acute inflammation and the localization in the lung. Guanidinium-, acetylated-, zwitterionic-, hydroxylated-, PEGylated-, carboxylated- and sulfated-F-PLNPs were synthesized from aminated-F-PLNP. The primary particle sizes were identical, but the hydrodynamic sizes ranged from 210 to 345 nm. Following surface functionalization, the F-PLNPs showed diverse zeta potentials from -41.2 to 31.0 mV, and each F-PLNP showed a single, narrow peak. Pharyngeal aspiration with these eight types of F-PLNPs into rats produced diverse acute lung inflammation, with zeta potentials of the F-PLNPs showing excellent correlation with acute pulmonary inflammation parameters including the percentage of polymorphonuclear leukocytes (R(2) = 0.90, p < 0.0001) and the levels of interleukin-1ß (R(2) = 0.83, p < 0.0001) and of cytokine-induced neutrophil chemoattractant-3 (R(2) = 0.86, p < 0.0001). These results imply that surface charge is a key factor influencing lung inflammation by functionalized polymeric NPs, which further confirms and extends the surface charge paradigm that we reported for pristine metal oxide NPs. This demonstrates that the surface charge paradigm is a valuable tool to predict the toxicity of NPs.

Comparison of cellular toxicity caused by ambient ultrafine particles and engineered metal oxide nanoparticles.

OBJECTIVE: The development of nanotechnology has spurred concerns about the health effects of exposure to nanoparticles (NPs) and ultrafine particles (UFPs). Toxicological data on NPs and UFPs may provide evidence to support the development of regulations to reduce the risk of particle exposure. We tried to provide fundamental data to determine differences in cytotoxicity induced by ambient UFPs and engineered metal oxide NPs (ZnO, NiO, and CeO2). METHODS: UFPs were sampled by using of a nano micro-orifice uniform deposit impactor. Physicochemical characterization of the UFPs and nano metal oxide particles were studied by scanning electron microscopy and transmission electron microscopy. Cellular toxicity induced by the different particles was assessed by using of comprehensive approaches and compared after A549 cells were exposured to the particles. RESULTS: All of the measured particles could damage A549 cells at concentrations ranging from 25 to 200 µg/mL. The lowest survival ratio and the highest lactate dehydrogenase level were caused by nano-ZnO particles, but the highest levels of intracellular reactive oxygen species (ROS) and percentages of apoptosis were observed in cells treated with the soluble fraction of ambient fine particles (PM1.8) at 200 µg/mL. Relatively high concentrations of anthropogenic metals, including Zn, Ni, Fe, and Cu, may be responsible for the higher toxicity of fine ambient particles compared with the ambient coarse particles and UFPs. The selected heavy metals (Zn, Ni, Fe, and Cu) were found to be located in the perinuclear and cytoplasmic areas of A549 cells. The distribution pattern of metals from ambient particles showed that distributions of the metals in A549 cells were not uniform and followed the pattern Cu>Zn>Fe>Ni, suggesting that Cu was absorbed by A549 cells more easily than the other metals. CONCLUSIONS: Metal nanoparticles oxides and UFPs at low concentration could damage to cells, but the manufactured metal oxide nanoparticles are not highly toxic to lung cells compared to environmental particles. The local concentration effect of heavy metals in A549 cells, as well as the induction of oxidative stress by the particles, may be responsible for the damage observed to the cells.

Toxicology of ZnO and TiO2 nanoparticles on hepatocytes: impact on metabolism and bioenergetics.

BACKGROUND AND AIM: Zinc oxide (ZnO) and titanium dioxide (TiO2) nanomaterials (NMs) are used in many consumer products, including foodstuffs. Ingested and inhaled NM can reach the liver. Whilst their effects on inflammation, cytotoxicity, genotoxicity and mitochondrial function have been explored, no work has been reported on their impact on liver intermediary metabolism. Our aim was to assess the effects of sub-lethal doses of these materials on hepatocyte intermediary metabolism. MATERIAL AND METHODS: After characterisation, ZnO and TiO2 NM were used to treat C3A cells for 4 hours at concentrations ranging between 0 and 10 µg/cm(2), well below their EC50, before the assessment of (i) glucose production and glycolysis from endogenous glycogen and (ii) gluconeogenesis and glycolysis from lactate and pyruvate (LP). Mitochondrial membrane potential was assessed using JC-10 after 0-40 µg/cm(2) ZnO. qRT-PCR was used to assess phosphoenolpyruvate carboxykinase (PEPCK) mRNA expression. Dihydroethidium (DHE) staining and FACS were used to assess intracellular reactive oxygen species (ROS) concentration. RESULTS: Treatment of cells with ZnO, but not TiO2, depressed mitochondrial membrane potential, leading to a dose-dependent increase in glycogen breakdown by up to 430%, with an increase of both glycolysis and glucose release. Interestingly, gluconeogenesis from LP was also increased, up to 10-fold and correlated with a 420% increase in the PEPCK mRNA expression, the enzyme controlling gluconeogenesis from LP. An intracellular increase of ROS production after ZnO treatment could explain these effects. CONCLUSION: At sub-lethal concentrations, ZnO nanoparticles dramatically increased both gluconeogenesis and glycogenolysis, which warrants further in vivo studies.

Chronic kidney disease rather than illness severity predicts medium- to long-term mortality and renal outcome after acute kidney injury.

BACKGROUND: Acute kidney injury (AKI) requiring renal replacement therapy (RRT) continues to be associated with a hospital mortality of ∼50%. Longer-term outcomes have been less well studied. We sought to determine the influence of ventilation and of underlying chronic kidney disease (CKD) on medium and longterm mortality and renal outcomes. METHODS: All patients requiring RRT for AKI in south west Scotland between 1 January 1994 and 31 December 2005 were followed prospectively. Survival of patients who were and were not ventilated and of those with and without underlying CKD was compared by odds ratio (OR), adjusting for age and sex. Renal outcomes were determined by interrogation of our biochemistry database. RESULTS: Three hundred and ninety-six patients with AKI received RRT. One hundred and seventy-six (44%) were ventilated and 98 (25%) had underlying CKD. Patients who required ventilation had a significantly worse 90-day survival than those who did not (OR 2.10 for death; 95% CI 1.34, 3.29) whereas underlying CKD did not predict such an early adverse outcome (OR 1.49; 95% CI 0.89, 2.50). By 5 years, patients who had been ventilated during the acute illness were no longer at increased risk (OR 0.79; 95% CI 0.38, 1.62) whereas the adverse effect of underlying CKD was statistically significant (OR 6.05; 95% CI 2.23, 16.5). Underlying CKD was also a strong predictor of the need for RRT during follow-up. CONCLUSION: In an unselected series of patients with AKI requiring RRT, underlying CKD rather than illness severity predicted medium- to long-term mortality.