Transcriptomic Remodelling of Fetal Endothelial Cells During Establishment of Inflammatory Memory.

Inflammatory memory involves the molecular and cellular 'reprogramming' of innate immune cells following exogenous stimuli, leading to non-specific protection against subsequent pathogen exposure. This phenomenon has now also been described in non-hematopoietic cells, such as human fetal and adult endothelial cells. In this study we mapped the cell-specific DNA methylation profile and the transcriptomic remodelling during the establishment of inflammatory memory in two distinct fetal endothelial cell types - a progenitor cell (ECFC) and a differentiated cell (HUVEC) population. We show that both cell types have a core transcriptional response to an initial exposure to a viral-like ligand, Poly(I:C), characterised by interferon responsive genes. There was also an ECFC specific response, marked by the transcription factor ELF1, suggesting a non-canonical viral response pathway in progenitor endothelial cells. Next, we show that both ECFCs and HUVECs establish memory in response to an initial viral exposure, resulting in an altered subsequent response to lipopolysaccharide. While the capacity to train or tolerize the induction of specific sets of genes was similar between the two cell types, the progenitor ECFCs show a higher capacity to establish memory. Among tolerized cellular pathways are those involved in endothelial barrier establishment and leukocyte migration, both important for regulating systemic immune-endothelial cell interactions. These findings suggest that the capacity for inflammatory memory may be a common trait across different endothelial cell types but also indicate that the specific downstream targets may vary by developmental stage.

Australian National Enterovirus Reference Laboratory annual report, 2015.

Australia conducts surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years as recommended by the World Health Organization (WHO) as the main method to monitor its polio-free status. Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2015, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.2 non-polio AFP cases per 100,000 children, meeting the WHO performance criterion for a sensitive surveillance system. Two non-polio enteroviruses, enterovirus A71 and coxsackievirus B3, were identified from clinical specimens collected from AFP cases. Australia complements the clinical surveillance program with enterovirus and environmental surveillance for poliovirus. Two Sabin-like polioviruses were isolated from sewage collected in Melbourne in 2015, which would have been imported from a country that uses the oral polio vaccine. The global eradication of wild poliovirus type 2 was certified in 2015 and Sabin poliovirus type 2 will be withdrawn from oral polio vaccine in April 2016. Laboratory containment of all remaining wild and vaccine strains of poliovirus type 2 will occur in 2016 and the National Enterovirus Reference Laboratory was designated as a polio essential facility. Globally, in 2015, 74 cases of polio were reported, only in the two remaining countries endemic for wild poliovirus: Afghanistan and Pakistan. This is the lowest number reported since the global polio eradication program was initiated.

Australian National Enterovirus Reference Laboratory annual report, 2014.

Following the World Health Organization (WHO) recommendation, Australia conducts surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age as the main method to monitor its polio-free status. Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2014, no cases of poliomyelitis were reported from clinical surveillance and Australia reported 1.4 non-polio AFP cases per 100,000 children, meeting the WHO performance criterion for a sensitive surveillance system. Non-polio enteroviruses can also be associated with AFP and enterovirus A71 and echovirus types 6 and 7 were identified from clinical specimens from cases of AFP. Globally, 359 cases of polio were reported in 2014, with the 3 endemic countries, Afghanistan, Nigeria and Pakistan, accounting for 95% of the cases. In May 2014, the WHO declared the international spread of wild poliovirus to be a public health emergency of international concern and has since maintained recommendations for polio vaccination of travellers from countries reporting cases of wild polio.

Annual report of the Australian National Enterovirus Reference Laboratory 2010-2011.

Australia conducts clinical surveillance for cases of polio-like illness in children in accordance with the World Health Organization (WHO) recommended surveillance criteria for acute flaccid paralysis (AFP). AFP cases are ascertained either by clinicians notifying the Australian Paediatric Surveillance Unit or designated nurses enrolling cases as part of the Paediatric Active Enhanced Disease Surveillance system at four sentinel tertiary paediatric hospitals. The National Enterovirus Reference Laboratory (NERL), formerly the National Poliovirus Reference Laboratory, is accredited by the World Health Organization (WHO) for the testing of faecal specimens from cases of AFP and operates as a Poliovirus Regional Reference Laboratory for the Western Pacific Region. In 2010 and 2011, for the 3rd and 4th consecutive years, Australia met the WHO AFP surveillance performance indicator. This is indicative of a sensitive surveillance system capable of detecting an imported case of polio in children. However, the faecal collection rate for the virological investigation of AFP cases was below the WHO surveillance performance indicator in both years and represented a gap in Australia's polio surveillance. Enterovirus and environmental surveillance were established in Australia as virological surveillance to complement the clinical surveillance schemes. No poliovirus was detected by the clinical or virological surveillance schemes in 2010 or 2011 and Australia maintained its polio-free status. India was declared polio-free in January 2012, a significant step towards global polio eradication, leaving Afghanistan, Nigeria and Pakistan as the remaining countries endemic for wild poliovirus.

Annual report of the Australian National Enterovirus Reference Laboratory 2012.

In 2012 no cases of poliomyelitis were reported through clinical surveillance in Australia, and poliovirus was not detected through virological surveillance. Australia conducts surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years as the main mechanism to monitor its polio-free status in accordance with World Health Organization (WHO) recommendations. Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System. In 2012 Australia reported 1.2 non-polio AFP cases per 100,000 children, meeting the WHO performance criterion for a sensitive system for the fifth year in a row. However the faecal specimen collection rate from AFP cases was 29%, which was well below the WHO target of 80%. Virological surveillance for poliovirus consists of two components. Firstly, the Enterovirus Reference Laboratory Network of Australia (ERLNA) reports on the typing of enteroviruses detected in or isolated from clinical specimens. Secondly, environmental surveillance is conducted at sentinel sites. These surveillance systems are co-ordinated by the National Enterovirus Reference Laboratory (NERL).

Potential for the Australian and New Zealand paediatric intensive care registry to enhance acute flaccid paralysis surveillance in Australia: a data-linkage study.

BACKGROUND: Australia uses acute flaccid paralysis (AFP) surveillance to monitor its polio-free status. The World Health Organization criterion for a sensitive AFP surveillance system is the annual detection of at least one non-polio AFP case per 100,000 children aged less than 15 years, a target Australia has not consistently achieved. Children exhibiting AFP are likely to be hospitalised and may be admitted to an intensive care unit. This provides a potential opportunity for active AFP surveillance. METHODS: A data-linkage study for the period from 1 January 2005 to 31 December 2008 compared 165 non-polio AFP cases classified by the Polio Expert Panel with 880 acute neurological presentations potentially compatible with AFP documented in the Australian and New Zealand Paediatric Intensive Care (ANZPIC) Registry. RESULTS: Forty-two (25%) AFP cases classified by the Polio Expert Panel were matched to case records in the ANZPIC Registry. Of these, nineteen (45%) cases were classified as Guillain-Barré syndrome on both registries. Ten additional Guillain-Barré syndrome cases recorded in the ANZPIC Registry were not notified to the national AFP surveillance system. CONCLUSIONS: The identification of a further ten AFP cases supports inclusion of intensive care units in national AFP surveillance, particularly specialist paediatric intensive care units, to identify AFP cases that may not otherwise be reported to the national surveillance system.

Jungle yellow fever mosquito studies in Trinidad: 1954

Eruption of a yellow fever epizootic/epidemic was signalled by the Trinidad Regional Virus Laboratory with the isolation of virus from the first human case in April 1954. Mosquito investigations were conducted in three major areas. Descriptions are provided of the forest environments, the mosquito collection techniques, transportation logistics, mosquito identification and isolation of viruses. An addendum briefly reviews subsequent yellow fever outbreaks in the island, addresses the question of origin and provides recommendations for the future.

Yellow fever: evolution of ideas concerned with demonstrating the natural occurrence of transovarial transmission of virus in mosquitoes

In November 1978, an outbreak of disease among wild simians of south Trinidad alerted officials to the presence of jungle yellow fever (YF). Learning of the epizootic and having recently been involved in studies establishing transovarial transmission (TOT) of YF virus, this laboratory urged Trinidad workers to undertake demonstration of the natural occurence of TOT in the forest environment. The idea as well as a suggested procedural protocol involving vector mosquitoes (Haemagogus spp.) was accepted and acted upon. Initial procedures called for undertaking studies in areas where sick or dead monkeys were observed, collecting larvae as well as adult female Haemagogus from which eggs were to be obtained, hatched, and the ensuing larvae reared to adults and tested for virus. As time passed, various problems were encountered necessitating changes in field procedures. The most important was the substitution of ovitraps for adult female collections as a source of eggs. By 1980, the epizootic/epidemic was waning. Funding for additional studies became available from Canadian sources and the author was invited to be a consultant. Field and laboratory procedures were again modified and streamlined. Described are: (1) choice of field sites, (2) choice of ovitraps, (3) siting of ovitraps, (4) frequency of collections, (5) laboratory procedures, (6) search for male Haemagogus and female ovipositing activity, and (7) duration of a TOT study and conclusions.