are integral building blocks in the cellular membranes of animals and have important functions in signaling, regulation, and metabolism ( 1 ). To date, the majority of studies have focused on cholesterol, the most abundant sterol in mammals that serves as both a precursor and product to a host of important molecules, including steroid hormones, bile acids, oxysterols, and intermediates in the cholesterol biosynthetic pathway ( 2-4 ). Although cholesterol has gained signifi cant notoriety due to the compound's negative impact on health, other sterols, such as plant-derived phytosterols, are thought to offer potential human health benefi ts by lowering circulating cholesterol levels ( 5, 6 ).Novel functions for sterols and secosteroids continue to be identifi ed. For example, the cholesterol metabolites 25-hydroxycholesterol and 7 ␣ ,25-dihydroxycholesterol have recently been shown to play important signaling roles in the immune system ( 7-9 ). 24-hydroxycholesterol has been shown to be involved in cholesterol turnover in the brain, and it plays a role in memory ( 10 ) and glucose metabolism ( 11 ). Alterations in vitamin D levels arising from the lack of sun exposure and/or use of sunscreen are postulated to have a negative impact on health ( 12, 13 ). These fi ndings together with the large number of sterols suggest that there may be undiscovered roles for sterols in biology, and they highlight the need for continued research into the biochemical pathways associated with these compounds.The extraction and analysis of sterols in human plasma present a unique challenge due to their virtual insolubility, sequestration within lipoproteins, and dramatic differences in the levels of individual sterols. Cholesterol is the most abundant sterol, with circulating levels on the order of 1 to 3 mg/ml, whereas 25-hydroxycholesterol is a millionfold less abundant at 1 to 3 ng/ml ( 14, 15 ). The circulating form of sterols in humans is primarily as steryl esters, in which a fatty acid is esterifi ed to carbon 3 of the sterol; Abstract We describe the development of a method for the extraction and analysis of 62 sterols, oxysterols, and secosteroids from human plasma using a combination of HPLC-MS and GC-MS. Deuterated standards are added to 200 l of human plasma. Bulk lipids are extracted with methanol:dichloromethane, the sample is hydrolyzed using a novel procedure, and sterols and secosteroids are isolated using solid-phase extraction (SPE). Compounds are resolved on C 18 core-shell HPLC columns and by GC. Sterols and oxysterols are measured using triple quadrupole mass spectrometers, and lathosterol is measured using GC-MS. Detection for each compound measured by HPLC-MS was ʜ 1 ng/ml of plasma. Extraction effi ciency was between 85 and 110%; day-to-day variability showed a relative standard error of <10%. Numerous oxysterols were detected, including the side chain oxysterols 22-, 24-, 25-, and 27-hydroxycholesterol, as well as ring-structure oxysterols 7 ␣ -and 4 ␤ -hydroxycholesterol. Intermediates from the cholesterol b...

The PGD test detected bacterial contamination in 1:3069 (9 of 27,620) doses released as negative by prestorage culture in PLTs as young as 3 days old. Three contaminated doses, two clinically insignificant, had nonreactive PGD tests, while 0.51% of tests were false positives. Application of this test on day of issue can interdict contaminated units and prevent transfusion reactions.

People with HIV have higher rates of certain comorbidities, particularly cardiovascular disease and some malignancies, than people without HIV. As somatic mutations associated with agerelated clonal haematopoiesis (CH) are linked to similar comorbidities in the general population, we hypothesized that CH may be more prevalent in people with HIV. To address this issue, we established a prospective cohort study recruiting 220 HIV-positive and 226 HIVnegative participants aged 55 years or older in Australia. Demographic characteristics, clinical data and peripheral blood were collected to assess for the presence of CH mutations and identify potential risk factors for and clinical sequelae of CH. Investigators testing for CH were blinded to participants' HIV status. In total, 132 CH mutations were identified in 99 (22.2%) of 446 participants. CH was more prevalent in HIV-positive participants than HIV-negative participants (27.7% vs. 16.8%, p =0.006), overall and across all age groups. HIV infection was associated with an increased odds of having CH (adjusted odds ratio 2.10, 95% confidence interval 1.30-3.38, p=0.002). The most common genes mutated were DNMT3A (48.5%), TET2 (20.5%) and ASXL1 (11.4%). CH and HIV infection were independently associated with increases in blood parameters and biomarkers associated with inflammation. These data suggest a selective advantage for the emergence of CH in the context of chronic infection and inflammation related to HIV infection.

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ObjectiveThe aim of the study was to describe the use of nonoccupational postexposure prophylaxis (NPEP) in Australia, and to estimate the number of HIV infections that its use prevented. MethodsWe conducted a population-based observational cohort study of people who presented to antiretroviral prescribers in Eastern Australia, and reported a high-risk nonoccupational exposure to HIV, in 1998HIV, in -2004 Prescribers collected data at baseline, 4 weeks and 6 months. Data collected included details of HIV exposure, drug regimens and HIV serostatus. ResultsThe great majority of the 1601 participants were male (95%) and presented after male homosexual exposure (87%). Only 32% of exposures were to HIV-positive sources. Two antiretroviral drugs were prescribed after 48% of events, and three or more drugs after 52% of events. The median time to receipt of NPEP was 23 h. Side effects were reported by 66% of participants. No case of NPEP failure in an adherent individual was identified. It was estimated that 0.9-9.2 HIV infections had been prevented. This compared with a total of 1138 newly acquired HIV infections notified in the geographical area covered by the study. ConclusionsIn Australia, NPEP has been widely prescribed and is mainly targeted at high-risk exposures. Although there were no identified failures of NPEP, it is likely that only a small proportion of new HIV infections in the study area were prevented. NPEP may be a valuable preventive intervention for an individual, but it can only play a minor role in HIV prevention at the population level unless targeting can be further improved. IntroductionNonoccupational postexposure prophylaxis (NPEP) remains a controversial method of HIV infection prevention. Only a relatively small number of countries have published guidelines recommending its use [1][2][3][4][5]. Available guidelines are inconsistent with respect to the recommended antiretroviral regimen, including the choice of two vs. three drugs, the type of exposures that are eligible, and whether or not the source is required to be HIV positive [6,7].Implementation of NPEP on a more extensive scale has been impeded by concerns about its efficacy, inappropriate use, toxicities and potential undermining of behavioural HIV infection prevention strategies [8][9][10].In Australia, guidelines recommending the use of NPEP after high-risk exposures were released in New South Wales (NSW) in 1998 [4], and national guidelines were published in 2001 [11]. The guidelines have been widely promoted among doctors and among people at high risk of HIV infection, predominantly homosexual men. Levels of awareness of its availability in this population are relatively high [12]. In this report, we describe the Australian experience of NPEP and examine its public health impact in a setting where its use is well accepted and relatively widespread. Participants were eligible for this study whether or not they elected to take NPEP. The national guidelines recommended two-antiretroviral-drug NPEP in most situations. Under the guid...

As treatment improves, people living with HIV (PLWHIV) can now expect to live longer, which means that the foci of HIV-related care for them and their medical practitioners continue to change. With an increasingly older cohort of patients with HIV infection, practitioners' key considerations are shifting from issues of acute treatment and patient survival to multiple comorbidities, toxicities associated with chronic therapy, and ongoing health maintenance. Within this context, this paper explores the current standard of practice for the management of HIV infection in Australia. We surveyed 56 Australian practitioners currently involved in managing HIV infection: 'HIV section 100' (HIV therapy-prescribing) general practitioners (s100 GPs; n = 26), sexual health physicians (SHPs; n = 24) and hospital-based physicians (HBPs; n = 6). Survey results for practice approaches and challenges were broadly consistent across the three practitioner specialties, apart from a few key areas. s100 GPs reported less prophylaxis use among patients whom they deemed at risk of HIV infection in comparison with SHPs, which may reflect differences in patient populations. Further, a higher proportion of s100 GPs nominated older HIV treatment regimens as their preferred therapy choices compared with the other specialties. In contrast with SHPs, s100 GPs were less likely to switch HIV therapies to simplify the treatment protocol, and to immediately initiate treatment upon patient request in those newly diagnosed with HIV infection. Considerably lower levels of satisfaction with current HIV practice guidelines were also reported by s100 GPs. It appears that greater support for s100 GPs may be needed to address these identified challenges and enhance approaches to HIV practice. Across all specialties, increasing access to mental health services for patients with HIV infection was reported as a key management issue. A renewed focus on providing improved mental health and wellbeing supports is recommended, particularly in the face of an ageing HIV-infected population.

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Background Synovial tissue from patients with osteoarthritis (OA) demonstrated synovial inflammation (Benito et al.). We hypothesized that a single intraarticular (IA) injection of an anti-TNF drug would result in decreased inflammatory cell infiltration and ultimately reduce articular injury. Objectives Evaluate the short-term benefit of IA anti-TNF therapy in knee osteoarthritis. Methods This study was a single center, 2:1:1 [INF:MP:P] randomization, double-blind, placebo-controlled treatment of knee OA with IA treatments of infliximab (INF) 100 mg, methylprednisolone (MP) 80 mg or saline (P) on Day 0. Subjects (n=16) had to have knee pain and show minimal to moderate osteoarthritic change on plain radiographs. Closed needle synovial biopsies (Bx) were obtained on Days 0 and 28. Total WOMAC was determined at Days 0, 14, 28 and 56. Bivariate associations were investigated using logistic and linear regression. Results Total WOMAC score improved significantly only for Group INF, comparing baseline to Day 56 (p<0.05) (See Fig). Overall 6 of 16 subjects had high levels of synovial cellularity (level 1-2, scale 0-3) at baseline, 4 of whom were in Group INF. These 4 subjects had the greatest degree of improvement in total WOMAC. Baseline CRP levels (baseline values =0.57-1.21 mg/dl) correlated with total WOMAC scores and did predict improvement (p=0.07). Baseline knee MRI grade (0-5) correlated with baseline total WOMAC but did not predict improvement. Circulating INF levels showed a maximum median concentration at Day 4 (3.70 ug/ml; range 0.56-11.27); two subjects found to have anti-INF antibodies by Day 14 showed reduced levels of INF at Day 14 (median 0.05 ug/ml), compared to the remaining 6 subjects at Day 14 (2.67 ug/ml; range 0.56-3.77). Synovial tissue showed reduction in two markers: CD68 macrophages (3 of 7 Bx INF Group) and CD54 ICAM-1 (2 of 4 Bx MP Group). Flow cytometric studies of PBMCs at Days 0 and 28 showed reduced numbers of INF subjects with TNF-expressing CD20 B cells and CD14 monocytes at Day 28. Conclusions In this pilot study the Total WOMAC score showed significant improvement by Day 56 (p<0.05) for the INF group. Baseline synovial cellularity and CRP also correlated with improvement. Anti-TNF IA therapy offers promise as symptomatic therapy. References Benito MJ, et al. Synovial tissue inflammation in early and late osteoarthritis. Ann Rheum Dis 2005;64:1263-7. Disclosure of Interest H. Lindsley Grant/Research support from: Janssen, J. Schue: None Declared, O. Tawfik: None Declared, R. Bolce: None Declared, D. Smith: None Declared, G. Hinson: None Declared, J. Wick: None Declared