Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium

The recognition of the potential efficacy of plasmid DNA (pDNA) molecules as vectors in the treatment and prevention of emerging diseases has birthed the confidence to combat global pandemics. This is due to the close-to-zero safety concern associated with pDNA vectors compared to viral vectors in cell transfection and targeting. Considerable attention has been paid to the potential of pDNA vectors but comparatively less thought has been given to the practical challenges in producing large quantities to meet current rising demands. A pilot-scale fermentation scheme was developed by employing a stoichiometrically-designed growth medium whose exceptional plasmid yield performance was attested in a shake flask environment for pUC19 and pEGFP-N1 transformed into E. coliDH5α and E. coliJM109, respectively. Batch fermentation of E. coliDH5α-pUC19 employing the stoichiometric medium displayed a maximum plasmid volumetric and specific yield of 62.6 mg/L and 17.1 mg/g (mg plasmid/g dry cell weight), respectively. Fed-batch fermentation of E. coliDH5α-pUC19 on a glycerol substrate demonstrated one of the highest ever reported pilot-scale plasmid specific yield of 48.98 mg/g and a volumetric yield of 0.53 g/L. The attainment of high plasmid specific yields constitutes a decrease in plasmid manufacturing cost and enhances the effectiveness of downstream processes by reducing the proportion of intracellular impurities. The effect of step-rise temperature induction was also considered to maximize ColE1-origin plasmid replication.

Weight and weight change following breast cancer : evidence from a prospective, population-based, breast cancer cohort study

Background: While weight gain following breast cancer is considered common, results supporting these findings are dated. This work describes changes in body weight following breast cancer over 72 months, compares weight with normative data and explores whether weight changes over time are associated with personal, diagnostic, treatment or behavioral characteristics. Methods: A population-based sample of 287 Australian women diagnosed with early-stage invasive breast cancer was assessed prospectively at six, 12, 18 and 72 months post-surgery. Weight was clinically measured and linear mixed models were used to explore associations between weight and participant characteristics (collected via self-administered questionnaire). Those with BMI changes of one or more units were considered to have experienced clinically significant changes in weight. Results: More than half (57%) of participants were overweight or obese at 6 months post-surgery, and by 72 months post-surgery 68% of women were overweight or obese. Among those who gained more weight than age-matched norms, clinically significant weight gain between 6 and 18 months and 6 and 72 months post-surgery was observed in 24% and 39% of participants, respectively (median [range] weight gain: 3.9kg [2.0-11.3kg] and 5.2kg [0.6-28.7], respectively). Clinically-significant weight losses were observed in up to 24% of the sample (median [range] weight loss between 6 and 72 months post-surgery: -6.4kg [-1.9--24.6kg]). More extensive lymph node removal, being treated on the non-dominant side, receiving radiation therapy and lower physical activity levels at 6 months was associated with higher body weights post-breast cancer (group differences >3kg; all p<0.05). Conclusions: While average weight gain among breast cancer survivors in the long-term is small, subgroups of women experience greater gains linked with adverse health and above that experienced by age-matched counterparts. Weight change post-breast cancer is a contemporary public health issue and the integration of healthy weight education and support into standard breast cancer care has potential to significantly improve the length and quality of cancer survivorship.

Streamlined research funding using short proposals and accelerated peer review : an observational study

Background Despite the widely recognised importance of sustainable health care systems, health services research remains generally underfunded in Australia. The Australian Centre for Health Services Innovation (AusHSI) is funding health services research in the state of Queensland. AusHSI has developed a streamlined protocol for applying and awarding funding using a short proposal and accelerated peer review. Method An observational study of proposals for four health services research funding rounds from May 2012 to November 2013. A short proposal of less than 1,200 words was submitted using a secure web-based portal. The primary outcome measures are: time spent preparing proposals; a simplified scoring of grant proposals (reject, revise or accept for interview) by a scientific review committee; and progressing from submission to funding outcomes within eight weeks. Proposals outside of health services research were deemed ineligible. Results There were 228 eligible proposals across 4 funding rounds: from 29% to 79% were shortlisted and 9% to 32% were accepted for interview. Success rates increased from 6% (in 2012) to 16% (in 2013) of eligible proposals. Applicants were notified of the outcomes within two weeks from the interview; which was a maximum of eight weeks after the submission deadline. Applicants spent 7 days on average preparing their proposal. Applicants with a ranking of reject or revise received written feedback and suggested improvements for their proposals, and resubmissions composed one third of the 2013 rounds. Conclusions The AusHSI funding scheme is a streamlined application process that has simplified the process of allocating health services research funding for both applicants and peer reviewers. The AusHSI process has minimised the time from submission to notification of funding outcomes.

Development of health risk-based metrics for defining a heatwave : a time series study in Brisbane, Australia

Background: This study attempted to develop health risk-based metrics for defining a heatwave in Brisbane, Australia. Methods: Poisson generalised additive model was performed to assess the impact of heatwaves on mortality and emergency hospital admissions (EHAs) in Brisbane. Results: In general, the higher the intensity and the longer the duration of a heatwave, the greater the health impacts. There was no apparent difference in EHAs risk during different periods of a warm season. However, there was a greater risk of mortality in the second half of a warm season than that in the first half. While elderly (>75 years)were particularly vulnerable to both the EHA and mortality effects of a heatwave, the risk for EHAs also significantly increased for two other age groups (0-64 years and 65-74 years) during severe heatwaves. Different patterns between cardiorespiratory mortality and EHAs were observed. Based on these findings, we propose the use of a teiered heat warning system based on the health risk of heatwave. Conclusions: Health risk-based metrics are a useful tool for the development of local heatwave definitions. thsi tool may have significant implications for the assessment of heatwave-related health consequences and development of heatwave response plans and implementation strategies.

Exercise barriers self-efficacy : development and validation of a subcale for individuals with cancer-related lymphedema

Background No tool exists to measure self-efficacy for overcoming lymphedema-related exercise barriers in individuals with cancer-related lymphedema. However, an existing scale measures confidence to overcome general exercise barriers in cancer survivors. Therefore, the purpose of this study was to develop, validate and assess the reliability of a subscale, to be used in conjunction with the general barriers scale, for determining exercise barriers self-efficacy in individuals facing lymphedema-related exercise barriers. Methods A lymphedema-specific exercise barriers self-efficacy subscale was developed and validated using a cohort of 106 cancer survivors with cancer-related lymphedema, from Brisbane, Australia. An initial ten-item lymphedema-specific barrier subscale was developed and tested, with participant feedback and principal components analysis results used to guide development of the final version. Validity and test-retest reliability analyses were conducted on the final subscale. Results The final lymphedema-specific subscale contained five items. Principal components analysis revealed these items loaded highly (> 0.75) on a separate factor when tested with a well-established nine-item general barriers scale. The final five-item subscale demonstrated good construct and criterion validity, high internal consistency (Cronbach’s alpha=0.93) and test-retest reliability (ICC=0.67, p< 0.01). Conclusions A valid and reliable lymphedema-specific subscale has been developed to assess exercise barriers self-efficacy in individuals with cancer-related lymphedema. This scale can be used in conjunction with an existing general exercise barriers scale to enhance exercise adherence in this understudied patient group.

Food neophobia and its association with diet quality and weight in children aged 24 months : a cross sectional study

Background Food neophobia, the rejection of unknown or novel foods, may result in poor dietary patterns. This study investigates the cross-sectional relationship between neophobia in children aged 24 months and variety of fruit and vegetable consumption, intake of discretionary foods and weight. Methods Secondary analysis of data from 330 parents of children enrolled in the NOURISH RCT (control group only) and SAIDI studies was performed using data collected at child age 24 months. Neophobia was measured at 24 months using the Child Food Neophobia Scale (CFNS). The cross-sectional associations between total CFNS score and fruit and vegetable variety, discretionary food intake and BMI (Body Mass Index) Z-score were examined via multiple regression models; adjusting for significant covariates. Results At 24 months, more neophobic children were found to have lower variety of fruits (β=-0.16, p=0.003) and vegetables (β=-0.29, p<0.001) but have a greater proportion of daily energy from discretionary foods (β=0.11, p=0.04). There was no significant association between BMI Z-score and CFNS score. Conclusions Neophobia is associated with poorer dietary quality. Results highlight the need for interventions to (1) begin early to expose children to a wide variety of nutritious foods before neophobia peaks and (2) enable health professionals to educate parents on strategies to overcome neophobia.

Suppression of cluster ions during rapidly increasing particle number concentration events in the environment

We show that the cluster ion concentration (CIC) in the atmosphere is significantly suppressed during events that involve rapid increases in particle number concentration (PNC). Using a neutral cluster and air ion spectrometer, we investigated changes in CIC during three types of particle enhancement processes – new particle formation, a bushfire episode and an intense pyrotechnic display. In all three cases, the total CIC decreased with increasing PNC, with the rate of decrease being greater for negative CIC than positive. We attribute this to the greater mobility, and hence the higher attachment coefficient, of negative ions over positive ions in the air. During the pyrotechnic display, the rapid increase in PNC was sufficient to reduce the CIC of both polarities to zero. At the height of the display, the negative CIC stayed at zero for a full 10 min. Although the PNCs were not significantly different, the CIC during new particle formation did not decrease as much as during the bushfire episode and the pyrotechnic display. We suggest that the rate of increase of PNC, together with particle size, also play important roles in suppressing CIC in the atmosphere.

Australian stock indexes and the four-factor model

Stock indexes are passive 'value-weighted' portfolios and should not have alphas which are significantly different from zero. If an index produces an insignificant alphan, then significant alphas for equity funds using this index can be attributed solely to manager performance. However, recent literature sugests that US Stock indexes can demonstrate significant alphas, which ultimately raise questions regarding equity fund manager performance in both the US and abroad. in this paper, we employ the Carhart four-factor model and newly available Asian-Pacific risk factors to generate alphas and risk factor loadings for eight Australian stock indexes from January 2004 to December 2012. We ifnd that the initial full sample period analysis does not provide indication of significant alphas in the indexes examined. However, by carrying out 36-month rolling regressions, we discover at least four significant alphas in seven of the eight indexes and factor loading variability. As previously reported in the US, this paper confirms similar issues with the four-factor model using Australian stock indexes and performance benchmarking. In effectively measuring Australian equity fund manager performance, it is therefore essential to evaluate a fund's alpha and risk factors relative to the alpha and risk factors of the appropriate benchmark index.

Child health and the income gradient : evidence from Australia

The positive relationship between household income and child health is well documented in the child health literature but the precise mechanisms via which income generates better health and whether the income gradient is increasing in child age are not well understood. This paper presents new Australian evidence on the child health–income gradient. We use data from the Longitudinal Study of Australian Children (LSAC), which involved two waves of data collection for children born between March 2003 and February 2004 (B-Cohort: 0–3 years), and between March 1999 and February 2000 (K-Cohort: 4–7 years). This data set allows us to test the robustness of some of the findings of the influential studies of Case et al. [Case, A., Lubotsky, D., Paxson, C., 2002. Economic status and health in childhood: the origins of the gradient. The American Economic Review 92 (5) 1308–1344] and Currie and Stabile [Currie, J., Stabile, M., 2003. Socioeconomic status and child health: why is the relationship stronger for older children. The American Economic Review 93 (5) 1813–1823], and a recent study by Currie et al. [Currie, A., Shields, M.A., Price, S.W., 2007. The child health/family income gradient: evidence from England. Journal of Health Economics 26 (2) 213–232]. The richness of the LSAC data set also allows us to conduct further exploration of the determinants of child health. Our results reveal an increasing income gradient by child age using similar covariates to Case et al. [Case, A., Lubotsky, D., Paxson, C., 2002. Economic status and health in childhood: the origins of the gradient. The American Economic Review 92 (5) 1308–1344]. However, the income gradient disappears if we include a rich set of controls. Our results indicate that parental health and, in particular, the mother's health plays a significant role, reducing the income coefficient to zero; suggesting an underlying mechanism that can explain the observed relationship between child health and family income. Overall, our results for Australian children are similar to those produced by Propper et al. [Propper, C., Rigg, J., Burgess, S., 2007. Child health: evidence on the roles of family income and maternal mental health from a UK birth cohort. Health Economics 16 (11) 1245–1269] on their British child cohort.

Sealing the leak on classical NTRU signatures

Initial attempts to obtain lattice based signatures were closely related to reducing a vector modulo the fundamental parallelepiped of a secret basis (like GGH [9], or NTRUSign [12]). This approach leaked some information on the secret, namely the shape of the parallelepiped, which has been exploited on practical attacks [24]. NTRUSign was an extremely efficient scheme, and thus there has been a noticeable interest on developing countermeasures to the attacks, but with little success [6]. In [8] Gentry, Peikert and Vaikuntanathan proposed a randomized version of Babai’s nearest plane algorithm such that the distribution of a reduced vector modulo a secret parallelepiped only depended on the size of the base used. Using this algorithm and generating large, close to uniform, public keys they managed to get provably secure GGH-like lattice-based signatures. Recently, Stehlé and Steinfeld obtained a provably secure scheme very close to NTRUSign [26] (from a theoretical point of view). In this paper we present an alternative approach to seal the leak of NTRUSign. Instead of modifying the lattices and algorithms used, we do a classic leaky NTRUSign signature and hide it with gaussian noise using techniques present in Lyubashevky’s signatures. Our main contributions are thus a set of strong NTRUSign parameters, obtained by taking into account latest known attacks against the scheme, a statistical way to hide the leaky NTRU signature so that this particular instantiation of CVP-based signature scheme becomes zero-knowledge and secure against forgeries, based on the worst-case hardness of the O~(N1.5)-Shortest Independent Vector Problem over NTRU lattices. Finally, we give a set of concrete parameters to gauge the efficiency of the obtained signature scheme.

Pan-Plasmodium band sensitivity for Plasmodium falciparum detection in combination malaria rapid diagnostic tests and implications for clinical management

Background: Malaria rapid diagnostic tests (RDTs) are appropriate for case management, but persistent antigenaemia is a concern for HRP2-detecting RDTs in endemic areas. It has been suggested that pan-pLDH test bands on combination RDTs could be used to distinguish persistent antigenaemia from active Plasmodium falciparum infection, however this assumes all active infections produce positive results on both bands of RDTs, an assertion that has not been demonstrated. Methods: In this study, data generated during the WHO-FIND product testing programme for malaria RDTs was reviewed to investigate the reactivity of individual test bands against P. falciparum in 18 combination RDTs. Each product was tested against multiple wild-type P. falciparum only samples. Antigen levels were measured by quantitative ELISA for HRP2, pLDH and aldolase. Results: When tested against P. falciparum samples at 200 parasites/μL, 92% of RDTs were positive; 57% of these on both the P. falciparum and pan bands, while 43% were positive on the P. falciparum band only. There was a relationship between antigen concentration and band positivity; ≥4 ng/mL of HRP2 produced positive results in more than 95% of P. falciparum bands, while ≥45 ng/mL of pLDH was required for at least 90% of pan bands to be positive. Conclusions: In active P. falciparum infections it is common for combination RDTs to return a positive HRP2 band combined with a negative pan-pLDH band, and when both bands are positive, often the pan band is faint. Thus active infections could be missed if the presence of a HRP2 band in the absence of a pan band is interpreted as being caused solely by persistent antigenaemia.

Soft-switching single inductor current-fed push-pull converter for PV applications

A switching control strategy is proposed for single inductor current-fed push-pull converter with a secondary side active voltage doubler rectifier or a voltage rectifier used in photovoltaic (PV) grid interfacing. The proposed switching control strategy helps to turn-on and turn-off the primary side power switches with zero-voltage and zero-current switching. The operation of the push-pull converter is analyzed for two modes of operation. The feasibility of the proposed switching control strategy is validated using simulation and experimental results.

Néstor-Guillermo Progeria Syndrome: a biochemical insight into Barrier-to-Autointegration Factor 1, alanine 12 threonine mutation

Background Premature aging syndromes recapitulate many aspects of natural aging and provide an insight into this phenomenon at a molecular and cellular level. The progeria syndromes appear to cause rapid aging through disruption of normal nuclear structure. Recently, a coding mutation (c.34G > A [p.A12T]) in the Barrier to Autointegration Factor 1 (BANF1) gene was identified as the genetic basis of Néstor-Guillermo Progeria syndrome (NGPS). This mutation was described to cause instability in the BANF1 protein, causing a disruption of the nuclear envelope structure. Results Here we demonstrate that the BANF1 A12T protein is indeed correctly folded, stable and that the observed phenotype, is likely due to the disruption of the DNA binding surface of the A12T mutant. We demonstrate, using biochemical assays, that the BANF1 A12T protein is impaired in its ability to bind DNA while its interaction with nuclear envelope proteins is unperturbed. Consistent with this, we demonstrate that ectopic expression of the mutant protein induces the NGPS cellular phenotype, while the protein localizes normally to the nuclear envelope. Conclusions Our study clarifies the role of the A12T mutation in NGPS patients, which will be of importance for understanding the development of the disease.