THE EFFECT OF NUTRITION ON LONGEVITY AND FERTILITY IN DIVERSE DROSOPHILA: A TOR-MEDIATED PROCESS

Nutrition and growth are strongly linked, but not much is known about how nutrition leads to growth. To understand the connection between nutrition through diet, growth and/or proliferation, we need to study the phenotypes resulting from the activation and inhibition of central metabolic pathways. Here I use the model organism Drosophila melanogaster (D. mel.) and three non-model Drosophila species with different dietary needs, Drosophila guttifera (D. gut.), Drosophila deflecta (D. def.), and Drosophila tripunctata (D. tri.), to study the effects of dietary amino acid availability in these diverse flies on fecundity and longevity. In addition, I inhibited the Target of Rapamycin (TOR) pathway to test how inhibition interplays with the nutritional stimuli in these four fruit fly species. The hypothesis was that the inhibition of the TOR pathway would reverse the phenotypes observed under conditions of overfeeding. The results show that egg-lay rates increased with higher amino acid availability but decreased in response to TOR inhibition. The longevity data were more varied: most species experienced an increase in median lifespan in both genders with an increase in yeast, except for D. mel. females. When exposed to the TOR inhibitor rapamycin, most lifespans decreased, except for D. tri. The obtained data will benefit future studies of metabolism and will also help scientists to adopt non-model organisms into their labs to study the true complexity of pathway evolution across different species with diverse nutritional habits and needs

Fetal Programming in Rheumatoid Arthritis

__Abstract__ Rheumatoid arthritis (RA) is a chronic, systemic, inflammatory, autoimmune disease mainly affecting synovial tissues, which can lead to severe morbidity and progressive joint destruction resulting in deformations and disability. Other important outcomes include extra-articular features and comorbidities, like vasculitis, cardiac disease and infections. RA affects approximately 1% of the adult population and each year, 5 to 50 per 100.000 persons develop this condition. Women are affected two to three times more often than men. The age of onset in women is usually between 40 and 50 years, but it often affects women of childbearing age

The Making of Transgenic

The complex color patterns on the wings and body of Drosophila guttifera (D. guttifera) are emerging as model systems for studying evolutionary and developmental processes. Studies regarding these processes depend on overexpression and downregulation of developmental genes, which ultimately rely upon an effective transgenic system. Methods describing transgenesis in Drosophila melanogaster (D. melanogaster) have been reported in several studies, but they cannot be applied to D. guttifera due to the low egg production rate and the delicacy of the eggs. In this protocol, we describe extensively a comprehensive method used for generating transgenic D. guttifera. Using the protocol described here, we are able to establish transgenic lines, identifiable by the expression of enhanced green fluorescent protein (EGFP) in the eye disks of D. guttifera larvae. The entire procedure, from injection to screening for transgenic larvae, can be completed in approximately 30 days and should be relatively easy to adapt to other non-model Drosophila species, for which no white-eyed mutants exist

Assessing Pharmacodynamic Interactions in Mice Using the Multistate Tuberculosis Pharmacometric and General Pharmacodynamic Interaction Models

The aim of this study was to investigate pharmacodynamic (PD) interactions in mice infected with Mycobacterium tuberculosis using population pharmacokinetics (PKs), the Multistate Tuberculosis Pharmacometric (MTP) model, and the General Pharmacodynamic Interaction (GPDI) model. Rifampicin, isoniazid, ethambutol, or pyrazinamide were administered in monotherapy for 4 weeks. Rifampicin and isoniazid showed effects in monotherapy, whereas the animals became moribund after 7 days with ethambutol or pyrazinamide alone. No PD interactions were observed against fast-multiplying bacteria. Interactions between rifampicin and isoniazid on killing slow and non-multiplying bacteria were identified, which led to an increase of 0.86 log10 colony-forming unit (CFU)/lungs at 28 days after treatment compared to expected additivity (i.e., antagonism). An interaction between rifampicin and ethambutol on killing non-multiplying bacteria was quantified, which led to a decrease of 2.84 log10 CFU/lungs at 28 days after treatment (i.e., synergism). These results show the value of pharmacometrics to quantitatively assess PD interactions in preclinical tuberculosis drug development

PREDICT TB : Prevention of Resistance, Evaluation of Diagnostics and Intensified or Custom-made Treatment of Tuberculosis

Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) is still a global health problem of immense proportion. Indeed, TB is considered the second most frequent cause of infectious disease-related death worldwide (after HIV-AIDS). The World Health Organization (WHO) is increasing its efforts to reduce the global threat of TB. Nevertheless, the morbidity and mortality statistics for TB remain shocking. For example, the WHO estimates that the global burden of disease caused by TB in 2010 comprised 8.8 million incident cases, including 1.1 million deaths from TB among HIV-negative individuals, and an additional 0.35 million deaths from HIV-associated TB.1 Additional to these impressive figures is the underlying problem of latent TB infections, with as many as one-third of the world’s population being thought to be latently infected with this bacterium, an immense pool of potential patients around. Further, all current treatment programs tend to be specifically aimed towards the treatment of active TB infection, leaving the huge underlying problem of latent TB infections untouched, essentially meaning that the worldwide elimination of TB is still a distant dream

Rna in situ hybridization for detecting gene expression patterns in the abdomens and wings of drosophila species

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species

Designing “little worlds” in Walnut Park: How architects adopted an ethnographic case study on living with dementia

Understanding future users is recognised to be essential in design, yet also challenging. Often architects have no direct access to the experiences of others, like people with dementia. Case studies have been suggested as an adequate format to inform designers. This paper investigates the role of an ethnographic case study about a person living with dementia, as provided to an architectural firm designing a residential care facility. Interviews with the architects and an analysis of design materials reveal how they incorporated the case study in their ongoing design. Results indicate that the case study offered insight into users’ daily life and facilitated architects’ concept development. Architects’ resulting concept proved valuable to frame design decisions, while its visualisation played a significant role in internal and external communication. The study contributes to untangling important aspects in informing architects about future users and raises questions regarding researchers’ and designers’ roles in transferring knowledge

“Catch 22”: biosecurity awareness, interpretation and practice amongst poultry catchers

Campylobacter contamination of chicken on sale in the UK remains at high levels and has a substantial public health impact. This has prompted the application of many interventions in the supply chain, including enhanced biosecurity measures on-farm. Catching and thinning are acknowledged as threats to the maintenance of good biosecurity, yet the people employed to undertake this critical work (i.e. ‘catchers’) are a rarely studied group. This study uses a mixed methods approach to investigate catchers’ (n = 53) understanding of the biosecurity threats posed by the catching and thinning, and the barriers to good biosecurity practice. It interrogated the role of training in both the awareness and practice of good biosecurity. Awareness of lapses in biosecurity was assessed using a Watch-&-Click hazard awareness survey (n = 53). Qualitative interviews (n = 49 catchers, 5 farm managers) explored the understanding, experience and practice of catching and biosecurity. All of the catchers who took part in the Watch-&-Click study identified at least one of the biosecurity threats with 40% detecting all of the hazards. Those who had undergone training were significantly more likely to identify specific biosecurity threats and have a higher awareness score overall (48% compared to 9%, p = 0.03). Crucially, the individual and group interviews revealed the tensions between the high levels of biosecurity awareness evident from the survey and the reality of the routine practice of catching and thinning. Time pressures and a lack of equipment rather than a lack of knowledge appear a more fundamental cause of catcher-related biosecurity lapses. Our results reveal that catchers find themselves in a ‘catch-22′ situation in which mutually conflicting circumstances prevent simultaneous completion of their job and compliance with biosecurity standards