Limited impact of awareness-raising campaigns on hepatitis C testing practices among general practitioners.

The global hepatitis strategy calls for increased effort to diagnose those infected, with a target of 90% diagnosed by 2030. Scotland's Action Plan on Hepatitis C included awareness-raising campaigns, undertaken during 2008-2011, to promote testing by general practitioners. We examined hepatitis C virus (HCV) testing practice among general practitioners before and following these campaigns. Scottish general practitioners were surveyed, using Dillman's method, in 2007 and 2013; response rates were 69% and 60%, respectively. Most respondents offer testing when presented with a risk history (86% in 2007, 88% in 2013) but only one-fifth actively sought out risk factors (19% in 2007, 21% in 2013). Testing was reportedly always/almost always/usually offered to people who inject drugs (84% in 2007, 87% in 2013). Significant improvements in the offer of testing were reported in patients with abnormal LFTs (41% in 2007, 65% in 2013, P<.001) and who had received medical/dental treatment in high prevalence countries (14% in 2007, 24% in 2013, P=.001). In 2013, 25% of respondents had undertaken HCV-related continued professional development. This group was significantly more likely to actively seek out risk factors (P=.009) but only significantly more likely to offer a test to patients who had received medical/dental treatment in high prevalence countries (P=.001). Our findings suggest that government-led awareness raising campaigns have limited impact on general practitioners' testing practices. If the majority of the HCV-infected population are to be diagnosed, practitioner-based or physician-centred interventions should be considered alongside educational initiatives targeted at professionals.

First Report of a Peronospora Species on Sweet Basil in South Africa.

Sweet basil (Ocimum basilicum) is an herbaceous aromatic annual plant of the family Lamiaceae grown for its flavoring and fragrances that can be used fresh or dried. In South Africa, sweet basil is grown on a commercial scale. Downy mildew has recently been reported as one of the most destructive diseases of sweet basil in Switzerland, France, and Italy (1-3). The identity of the downy mildew species infecting sweet basil has been controversial and has been indicated as Peronospora lamii, a presumably undescribed (unnamed) Peronospora species, as well as a few species of which the status as distinct species is mostly unclear or doubtful (1). The distinction between P. lamii and the unnamed Peronospora species has been based on their sporangial dimensions, with P. lamii having sporangial dimensions with a length and width range of 16 to 26 × 15 to 23 µm (average 21 × 18 µm) and the unnamed Peronospora species having sporangial dimensions of 20 to 35 × 15 to 25 µm (average 28 × 22 µm) (1) or 23 to 36 × 18 to 29 µm (average 29 × 23 µm) (2). Additionally, internal transcribed spacer (ITS) sequence data has also been used to show that P. lamii and the unnamed Peronospora species on basil are not similar (1). In the Western Cape Province of South Africa, a sweet basil sample was received at the Stellenbosch University Plant Disease Clinic in 2005 from a grower in the region who experienced almost 50% crop failure under greenhouse-grown conditions. Initial symptoms were chlorotic leaves that subsequently developed a brown sporulation on the abaxial side. Microscopic observations of the brown sporulation were consistent with a Peronospora species. The sporangiophores branched two to five times with lengths ranging from 130 to 290 µm (average 194 µm). Sporangiophores terminated with dichotomously branched denticels bearing single detachable sporangia. Sporangia measured 26 to 34 × 20 to 28 µm (average 30 × 24 µm) and were elliptical and brown. The sporangia were similar in shape, color, and size range as that previously reported for a unnamed Peronospora species on sweet basil (1,2). Sequence analyses were also conducted on two isolates by first extracting DNA from spores that were washed from leaves using the Wizard SV genomic DNA purification system (Promega, Madison, WI), followed by polymerase chain reaction (PCR) amplification and sequencing of the ITS1, 5.8S, and ITS2 regions using primers ITS6 and ITS4 (4). The sequences of the two isolates were identical (GenBank Accession No. DQ479408). BLAST analyses of the sequences revealed a 99% similarity to the unnamed Peronospora species that was isolated from sweet basil in Switzerland and Italy (1). The sequences of the South African isolates only had low homology to P. lamii. To our knowledge, this is the first report of a Peronospora species on sweet basil in South Africa that on the basis of morphology and ITS sequence data is similar to the unnamed Peronospora species recently described in Switzerland and Italy on sweet basil (1). References: (1) L. Belbahri et al. Mycol. Res. 109:1276, 2005. (2) A. Garibaldi et al. Plant Dis. 88:312, 2004. (3) A. Garibaldi et al. Plant Dis. 89:683, 2005. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds., Academic Press, San Diego, 1990.

Effects of in vitro incorporation of cholesterol and cholesterol analogues into rat platelets.

Cholesterol and analogues of cholesterol bearing shorter side chains were incorporated into rat platelet membranes by incubation with sterol-rich liposomes in vitro. Cholesterol-enriched platelets showed increased aggregability to collagen compared with controls. Platelets containing the cholesterol analogues pregn-5-en-3 beta-ol and chol-5-en-3 beta-ol were even more sensitive to aggregation and could aggregate spontaneously on stirring. The size of the platelets containing pregn-5-en-3 beta-ol was markedly reduced when compared with controls in the scanning electron microscope. The results suggest that the sterol content and structure of the platelet membrane can have a critical role in maintaining the normal function of the cell.

Loss of factor VIII activity during storage in PVC containers due to adsorption.

Recombinant factor VIII concentrates are stable when administered in a reconstituted form according to the manufacturer's specifications, and undiluted via infusion with syringe mini-pumps. However many Haemophilia centres administer recombinant factor VIII further diluted in intravenous fluids for greater ease of administration. To investigate the stability of recombinant factor VIII during administration as a diluted infusion, reconstituted factor VIII was stored in polyvinylchloride (PVC) mini-bags undiluted (146 IU mL-1) and at factor VIII concentrations of 10 IU mL-1 and 2 IU mL-1. After 48 h of storage at room temperature in PVC mini-bags, the recoveries of factor VIII activity were 41.9% of the initial activity for the undiluted (146 IU mL-1) product and 43.7% of the initial activity for factor VIII diluted to 10 IU mL-1. For factor VIII diluted to 2 IU mL-1, the amount of factor VIII activity remaining at 48 h was only 1.8% of the initial activity. In contrast, 100% of factor VIII activity was recovered after 48 h when undiluted reconstituted product (146 IU mL-1) was stored in a syringe. To investigate the mechanism of factor VIII activity loss during storage, factor VIII samples collected after 0, 3 and 48 h of storage were analysed by immunoblotting with factor VIII antibodies. No evidence of factor VIII proteolytic degradation during storage was found, however, large amounts of factor VIII antigen were recovered from the empty PVC mini-bags following elution with denaturing detergent. We conclude that clinically significant losses of factor VIII activity occur during storage in PVC mini-bags and that the loss of activity is most likely due to protein adsorption onto the plastic surface. This loss of factor VIII activity during storage in PVC containers may substantially affect the safety and potential cost savings of administering recombinant factor VIII by continuous infusion.