Effect of vesicle size on tissue localization and immunogenicity of liposomal DNA vaccines

The formulation of plasmid DNA (pDNA) in cationic liposomes is a promising strategy to improve the potency of DNA vaccines. In this respect, physicochemical parameters such as liposome size may be important for their efficacy. The aim of the current study was to investigate the effect of vesicle size on the in vivo performance of liposomal pDNA vaccines after subcutaneous vaccination in mice. The tissue distribution of cationic liposomes of two sizes, 500 nm (PDI 0.6) and 140 nm (PDI 0.15), composed of egg PC, DOPE and DOTAP, with encapsulated OVA-encoding pDNA, was studied by using dual radiolabeled pDNA-liposomes. Their potency to elicit cellular and humoral immune responses was investigated upon application in a homologous and heterologous vaccination schedule with 3 week intervals. It was shown that encapsulation of pDNA into cationic lipsomes resulted in deposition at the site of injection, and strongest retention was observed at large vesicle size. The vaccination studies demonstrated a more robust induction of OVA-specific, functional CD8+ T-cells and higher antibody levels upon vaccination with small monodisperse pDNA-liposomes, as compared to large heterodisperse liposomes or naked pDNA. The introduction of a PEG-coating on the small cationic liposomes resulted in enhanced lymphatic drainage, but immune responses were not improved when compared to non-PEGylated liposomes. In conclusion, it was shown that the physicochemical properties of the liposomes are of crucial importance for their performance as pDNA vaccine carrier, and cationic charge and small size are favorable properties for subcutaneous DNA vaccination.

The effect of rock surface aspect on growth, size structure and competition in the lichen Rhizocarpon geographicum

Rhizocarpon geographicum is a crustose lichen found frequently on rock surfaces of southern aspect and less frequently on rock surfaces of northern aspect in Gwynedd, North Wales. This study tested the hypothesis that the radial growth of R. geographicum thalli predicts aspect distribution. Thalli of all sizes, however, exhibited significantly greater radial growth over 18 months on northwest compared with southeast facing surfaces. The hypothesis that a more intense competitive environment on northwest facing surfaces may explain the aspect distribution of R. geographicum was then tested. The size frequency distributions of thalli revealed a higher proportion of thalli in the smallest size class and a more restricted thallus size range on the northwest facing surfaces. In addition, thallus mortality appeared to be greater on northwest facing surfaces. Significantly more associated lichen species were present on rock surfaces of northern aspect at sites where R. geographicum was present. The mean frequency of the associated lichen species, however, was significantly lower on surfaces of northern aspect where R. geographicum was present. In addition, two common foliose species at these sites were demonstrated experimentally to overgrow thalli of R. geographicum. It is concluded that the growth of R. geographicum over the study period did not predict aspect distribution and that differences in the competitive environments on northwest and southeast surfaces may be an important factor determining aspect distribution. © 2002 Elsevier Science B.V. All rights reserved.

A CFD approach on the effect of particle size on char entrainment in bubbling fluidised bed reactors

The fluid – particle interaction inside a 41.7 mg s-1 fluidised bed reactor is modelled. Three char particles of sizes 500 µm, 250 µm, and 100 µm are injected into the fluidised bed and the momentum transport from the fluidising gas and fluidised sand is modelled. Due to the fluidising conditions and reactor design the char particles will either be entrained from the reactor or remain inside the bubbling bed. The particle size is the factor that differentiates the particle motion inside the reactor and their efficient entrainment out of it. A 3-Dimensional simulation has been performed with a completele revised momentum transport model for bubble three-phase flow according to the literature as an extension to the commercial finite volume code FLUENT 6.2.

The vesicle size of DDA:TDB liposomal adjuvants plays a role in the cell-mediated immune response but has no significant effect on antibody production

The use of cationic liposomes as experimental adjuvants for subunit peptide of protein vaccines is well documented. Recently the cationic liposome CAF01, composed of dimethyldioctadecylammonium (DDA) and trehalose dibehenate (TDB), has entered Phase I clinical trials for use in a tuberculosis (TB) vaccine. CAF01 liposomes are a heterogeneous population with a mean vesicle size of 500 nm; a strong retention of antigen at the injection site and a Th1-biassed immune response are noted. The purpose of this study was to investigate whether CAF01 liposomes of significantly different vesicle sizes exhibited altered pharmacokinetics in vivo and cellular uptake with activation in vitro. Furthermore, the immune response against the TB antigen Ag85B-ESAT-6 was followed when various sized CAF01 liposomes were used as vaccine adjuvants. The results showed no differences in vaccine (liposome or antigen) draining from the injection site, however, significant differences in the movement of liposomes to the popliteal lymph node were noted. Liposome uptake by THP-1 vitamin D3 stimulated macrophage-like cells did not show a liposome size-dependent pattern of uptake. Finally, whilst there were no significant differences in the IgG1/2 regardless of the liposome size used as a delivery vehicle for Ag85B-ESAT-6, vesicle size has a size dependent effect on cell proliferation and IL-10 production with larger liposomes (in excess of 2 µm) promoting the highest proliferation and lowest IL-10 responses, yet vesicles of ~500 nm promoting higher IFN-? cytokine production from splenocytes and higher IL-1ß at the site of injection.

Characterisation of polyethylene powders for rotational moulding and effect of powder size and shape on densification behaviour

The particle size, shape and distribution of a range of rotational moulding polyethylenes (PEs) ground to powder was investigated using a novel visual data acquisition and analysis system (TP Picture®), developed by Total Petrochemicals. Differences in the individual particle shape factors of the powder samples were observed and correlations with the grinding conditions were determined. When heated, the bubble dissolution behaviour of the same powders was investigated and the shape factor correlated with densification rate, bubble size and bubble distribution.

The effect of hook size on the catch rate and size of the Nile perch Lates niloticus (Linne) fishery in the Napoleon Gulf, Lake Victoria, Uganda

This study was undertaken in Napoleon gulf, Lake Victoria Uganda from July – December 2009. It was conducted in four landing sites; Bukaya (0.41103N, 33.19133E), Bugungu (0.40216N, 33.2028E), Busana (0.39062N, 33.25228E) and Kikondo (0.3995N, 33.21848E) all from Buikwe district (Formerly part of Mukono district). The main aim was to determine the effect of both hook size and bait type on the catch rate (mean weight) and size composition of Nile perch (Lates niloticus) (LINNE) fishery in the Napoleon Gulf, Lake Victoria. The main hook sizes investigated during the experiment were 7, 8, 9, 10, 11 and 12 that were dominantly used in harvesting Nile perch in Napoleon Gulf, Lake Victoria. In this study length, weight and bait type data were collected on site from each boat at that particular fishing spot; since most fishermen in the Napoleon Gulf could sell their fish immediately the catch is caught there and then. The results indicated a total of 873 Nile perch fish samples collected during the study. Statistical tests, descriptive statistics, regression and correlation were all carried out using the Statistical Package for the Social Sciences (SPSS) in addition to Microsoft excel. The bait types in the Gulf ranged from 5-10 cm Total length (TL) haplochromine, 24.5-27 cm TL Mormyrus kannume and 9-24 cm TL Clarias species. The bait types had a significant effect on the catch rate and also on the size composition the fish harvested measured as Total length (ANCOVA F=8.231; P<0.05) despite the fact that bait type had no influence on mean weight of fish captured (ANCOVA F=2.898; P>0.05). Hook sizes used by the fishers had a significant effect on the both the size (TL) composition (ANCOVA F=3.847; P<0.05) and the mean weight (ANCOVA F=4.599; P<0.005) of the Nile perch captured. Investigations indicated hook sizes seven (7) and eight (8) were the ones that harvested the Nile perch above the slot size of 50 cm total length. In general hook sizes indicated to be the main drive in the harvesting of the Nile perch though bait type also contributed toward that. Generally there is need for management to put a law in place on the minimum hook size to be used on the harvesting of the Nile perch and also monitored by the Fisheries Management as a regulatory measure. In addition to that aquaculture should be encouraged to farm the fish for bait at a higher scale in the region in order to avoid depleting the wild stocks already in danger of extinction. Through this kind of venture, both biodiversity conservation and environmental sustainability will be observed in the Lake Victoria basin.

The effect of Lampara net on the catch rates and size of the fishing harvest of Rastrineobola argentae (mukene) in selected parts of Lake Victoria, Uganda

This study was undertaken in Napoleon gulf, and part of the off shore area of Lake Victoria Uganda in the landing site of Lufu in Buvuma district in the month of October 2015 for three days of sampling. It was conducted in four landing sites; Busana, Kikondo from Buikwe district and Lufu landing site from Buvuma district. The main aim was to determine the effect of Lampara net on the catch rate and size of the Rastrineobola argentae (mukene) harvested on Lake Victoria using various mesh sizes. The study focused on the 5mm and 10 mm mesh sizes of the Lampara net. A total of 109 boats were sampled; from the 5 mm mesh sizes indicated catch rates of 78-200 kg/boat/day and yet for 10 mm mesh size was at 248 kg/boat day. Statistical tests were carried out on the these two mesh sizes using One way Anova and indicated in catches was (Anova F=7.476; P<0.05) and for the price values was (Anova F=5.488 ; P<0.05). This is an indication that despite the fact that the mukene fishery is on the increase, a time will come when it also be depleted so a need to use the rightful fishing gear of 10mm mesh size is advisable for the biodiversity conservation.

Treatment trials for neonatal seizures: the effect of design on sample size

Neonatal seizures are common in the neonatal intensive care unit. Clinicians treat these seizures with several anti-epileptic drugs (AEDs) to reduce seizures in a neonate. Current AEDs exhibit sub-optimal efficacy and several randomized control trials (RCT) of novel AEDs are planned. The aim of this study was to measure the influence of trial design on the required sample size of a RCT. We used seizure time courses from 41 term neonates with hypoxic ischaemic encephalopathy to build seizure treatment trial simulations. We used five outcome measures, three AED protocols, eight treatment delays from seizure onset (Td) and four levels of trial AED efficacy to simulate different RCTs. We performed power calculations for each RCT design and analysed the resultant sample size. We also assessed the rate of false positives, or placebo effect, in typical uncontrolled studies. We found that the false positive rate ranged from 5 to 85% of patients depending on RCT design. For controlled trials, the choice of outcome measure had the largest effect on sample size with median differences of 30.7 fold (IQR: 13.7–40.0) across a range of AED protocols, Td and trial AED efficacy (p<0.001). RCTs that compared the trial AED with positive controls required sample sizes with a median fold increase of 3.2 (IQR: 1.9–11.9; p<0.001). Delays in AED administration from seizure onset also increased the required sample size 2.1 fold (IQR: 1.7–2.9; p<0.001). Subgroup analysis showed that RCTs in neonates treated with hypothermia required a median fold increase in sample size of 2.6 (IQR: 2.4–3.0) compared to trials in normothermic neonates (p<0.001). These results show that RCT design has a profound influence on the required sample size. Trials that use a control group, appropriate outcome measure, and control for differences in Td between groups in analysis will be valid and minimise sample size.

Using a longitudinal model to estimate the effect of methicillin-resistant staphylococcus aureus infection on length of stay in an intensive care unit

Healthcare-associated methicillin-resistant Staphylococcus aureus(MRSA) infection may cause increased hospital stay or, sometimes, death. Quantifying this effect is complicated because it is a time-dependent exposure: infection may prolong hospital stay, while longer stays increase the risk of infection. We overcome these problems by using a multinomial longitudinal model for estimating the daily probability of death and discharge. We then extend the basic model to estimate how the effect of MRSA infection varies over time, and to quantify the number of excess ICU days due to infection. We find that infection decreases the relative risk of discharge (relative risk ratio = 0.68, 95% credible interval: 0.54, 0.82), but is only indirectly associated with increased mortality. An infection on the first day of admission resulted in a mean extra stay of 0.3 days (95% CI: 0.1, 0.5) for a patient with an APACHE II score of 10, and 1.2 days (95% CI: 0.5, 2.0) for a patient with an APACHE II score of 30. The decrease in the relative risk of discharge remained fairly constant with day of MRSA infection, but was slightly stronger closer to the start of infection. These results confirm the importance of MRSA infection in increasing ICU stay, but suggest that previous work may have systematically overestimated the effect size.