Evaluation of an in-house molecular HIV-1 test to assess mother-to-child HIV-1 transmission in Angola (the APEHC cohort)

Poster presented at the 17th Annual International Meeting of the Institute of Human Virology. Baltimore, 27-30 September 2015

Role of Serum Levels of Vascular Endothelial Growth Factor and Its Receptor in Retinopathy of Prematurity

Background: Retinopathy of prematurity (ROP) is a disorder of developing retina of low birth weight preterm infants which can lead to blindness. One theory attributes the fibrosis seen in ROP to deregulation of vascularization in the retina. Vascular endothelial growth factor (VEGF) is one of the important mediators involved in vascularization. Objectives: This study was carried out to assess the role of VEGF and its receptor in retinopathy of prematurity. Patients and Methods: Around 200 preterm infants born in SSK hospital were screened at 33 - 34 weeks. These babies were followed up according to the international classification of retinopathy of prematurity (ICROP) criteria. Those infants who developed ROP at 38 - 40 weeks were enrolled in group A while an equal number of infants who did not develop ROP were included in group B. Each group comprised of 30 subjects each. Venous sampling was carried out twice, once at 33 - 34 weeks and then again at 38 - 40 weeks. VEGF and VEGF-R2 were estimated by commercially available ELISA kits. Results: There was no statistically significant difference between the levels of VEGF and VEGF-R2 in both groups at first visit as well as the follow up visit. However, the intra-group difference was significant between the first and the final visit in VEGF and VEGF-R2 levels in the cases with ROP. In the control population, the VEGF levels were significantly lower in the follow up visit as compared to the initial visit. Conclusions: Our study demonstrates that a significant difference is seen in the serum VEGF and VRGF-R2 in the second visit of the infants with ROP demonstrating that VEGF might be responsible for the initiation and aggravation of ROP.

Evaluation of the Effect of Passive Smoking on Lactoferrin and AST on 12 - 15 Years Old Children and Adolescents

Background: Passive smokers are involuntarily exposed to cigarette or tobacco smoke and as known, inhalation of environmental tobacco smoke is a serious threat. There is little information about the effect of passive smoking on salivary markers and periodontal indices. Objectives: This study investigated the effect of passive smoking on lactoferrin and AST in 12 - 15 years old children and adolescents. Patients and Methods: This case-control analytic correlation type study with no-convenience random sampling method was performed on 160 children aged 12 - 15 who had smokers in their families. The eligible children were divided into two equal groups; 80 cot+ children as case group and 80 cot– children as control group, matched according to age, sex and plaque index. Plaque index was obtained from all subjects. 2 cc unstimulated salivary samples were collected by spitting method. The collected specimens were tested by lactoferrin and AST kits in biochemistry were measured on the day of sampling laboratory. Gingival index Loe and Silness (GI) and Probing Pocket Depth (PPD). Results: Mean and Standard Deviation of PPD and GI was 2.01 ± 0.077 and 1.53 ± 0.055 in experimental group and 1.93 ± 0.073 and 1.49 ± 0.046 in control group respectively (P < 0.001). The Mean and Standard Deviation parameters of lactoferrin and AST, in the experimental group was 38.66 ± 25.15 and 13.45 ± 6.33 and in the control group 10.18 ± 6.82 and 6.53 ± 2.65 group, respectively (P < 0.001). Conclusions: Passive smoking can be effective on inflammatory process of periodontal and salivary biomarkers related to inflammation. Lactoferrin was 11 - 104 in case group and 0.5 - 38 in control group. Aspartat aminotransferase in case group was 2.64 - 30.43 and in control group it was 2.16 - 12.02.

Veteranness : Representations of Combat-related PTSD in U.S. Popular Visual Media

Posttraumatic stress and PTSD are becoming familiar terms to refer to what we often call the invisible wounds of war, yet these are recent additions to a popular discourse in which images of and ideas about combat-affected veterans have long circulated. A legacy of ideas about combat veterans and war trauma thus intersects with more recent clinical information about PTSD to become part of a discourse of visual media that has defined and continues to redefine veteran for popular audiences. In this dissertation I examine realist combat veteran representations in selected films and other visual media from three periods: during and after World Wars I and II (James Allen from I Am a Fugitive from a Chain Gang, Fred Derry and Al Stephenson from The Best Years of Our Lives); after the Vietnam War (Michael from The Deer Hunter, Eriksson from Casualties of War), and post 9/11 (Will James from The Hurt Locker, a collection of veterans from Wartorn: 1861-2010.) Employing a theoretical framework informed by visual media studies, Barthes’ concept of myth, and Foucault’s concept ofdiscursive unity, I analyze how these veteran representations are endowed with PTSD symptom-like behaviors and responses that seem reasonable and natural within the narrative arc. I contend that veteran myths appear through each veteran representation as the narrative develops and resolves. I argue that these veteran myths are many and varied but that they crystallize in a dominant veteran discourse, a discursive unity that I term veteranness. I further argue that veteranness entangles discrete categories such as veteran, combat veteran, and PTSD with veteran myths, often tying dominant discourse about combat-related PTSD to outdated or outmoded notions that significantly affect our attitudes about and treatment of veterans. A basic premise of my research is that unless and until we learn about the lasting effects of the trauma inherent to combat, we hinder our ability to fulfill our responsibilities to war veterans. A society that limits its understanding of posttraumatic stress, PTSD and post-war experiences of actual veterans affected by war trauma to veteranness or veteran myths risks normalizing or naturalizing an unexamined set of sociocultural expectations of all veterans, rendering them voice-less, invisible, and, ultimately disposable.

Preparation and characterization of grafted nonwoven membranes for bioseparations

Protein purification plays a crucial role in biotechnology and biomanufacturing, where downstream unit operations account for 40%-80% of the overall costs. To overcome this issue, companies strive to simplify the separation process by reducing the number of steps and replacing expensive separation devices. In this context, commercially available polybutylene terephthalate (PBT) melt-blown nonwoven membranes have been developed as a novel disposable membrane chromatography support. The PBT nonwoven membrane is able to capture products and reduce contaminants by ion exchange chromatography. The PBT nonwoven membrane was modified by grafting a poly(glycidyl methacrylate) (GMA) layer by either photo-induced graft polymerization or heat induced graft polymerization. The epoxy groups of GMA monomer were subsequently converted into cation and anion exchangers by reaction with either sulfonic acid groups or diethylamine (DEA), respectively. Several parameters of the procedure were studied, especially the effect of (i) % weight gain and (ii) ligand density on the static protein binding capacity. Bovine Serum Albumin (BSA) and human Immunoglobulin G (hIgG) were utilized as model proteins in the anion and cation exchange studies. The performance of ion exchange PBT nonwovens by HIG was evaluated under flow conditions. The anion- and cation- exchange HIG PBT nonwovens were evaluated for their ability to selectively adsorb and elute BSA or hIgG from a mixture of proteins. Cation exchange nonwovens were not able to reach a good protein separation, whereas anion exchange HIG nonwovens were able to absorb and elute BSA with very high value of purity and yield, in only one step of purification.

The effect of sample and sample matrix on DNA processing: Mechanisms for the detection and management of inhibition in forensic samples

The presence of inhibitory substances in biological forensic samples has, and continues to affect the quality of the data generated following DNA typing processes. Although the chemistries used during the procedures have been enhanced to mitigate the effects of these deleterious compounds, some challenges remain. Inhibitors can be components of the samples, the substrate where samples were deposited or chemical(s) associated to the DNA purification step. Therefore, a thorough understanding of the extraction processes and their ability to handle the various types of inhibitory substances can help define the best analytical processing for any given sample. A series of experiments were conducted to establish the inhibition tolerance of quantification and amplification kits using common inhibitory substances in order to determine if current laboratory practices are optimal for identifying potential problems associated with inhibition. DART mass spectrometry was used to determine the amount of inhibitor carryover after sample purification, its correlation to the initial inhibitor input in the sample and the overall effect in the results. Finally, a novel alternative at gathering investigative leads from samples that would otherwise be ineffective for DNA typing due to the large amounts of inhibitory substances and/or environmental degradation was tested. This included generating data associated with microbial peak signatures to identify locations of clandestine human graves. Results demonstrate that the current methods for assessing inhibition are not necessarily accurate, as samples that appear inhibited in the quantification process can yield full DNA profiles, while those that do not indicate inhibition may suffer from lowered amplification efficiency or PCR artifacts. The extraction methods tested were able to remove >90% of the inhibitors from all samples with the exception of phenol, which was present in variable amounts whenever the organic extraction approach was utilized. Although the results attained suggested that most inhibitors produce minimal effect on downstream applications, analysts should practice caution when selecting the best extraction method for particular samples, as casework DNA samples are often present in small quantities and can contain an overwhelming amount of inhibitory substances.^

International flavours

It is an Olympic year and we have just witnessed the fantastic games hosted by Rio de Janeiro. Well done to team USA for winning the most medals overall but also well done to so many other nations and individuals who performed so well or were ambassadors in other ways. Teenage swimmer Yusra Mardini who swam for the refugee team and South Africa's Wayde van Niekerk who broke the longstanding 400 m record of Michael Johnson that has stood since 1999. Of course, we must mention sprinter Usain Bolt and swimmer Michael Phelps, who have now transcended superstar status and entered a new level of icon. My personal highlight was the sportsmanship witnessed in the 5000 m when American Abbey D’Agostino was accidentally felled by New Zealand runner Nikki Hamblin. D’Agostino helped Hamblin back to her feet but slumped to the track after realising her own injury. Hamblin helped her up and stayed with her so that both completed the race. The International Olympic Committee has awarded both with the prestigious Pierre de Coubertin award, also known as the International Fair Play Trophy. Fair play is of paramount importance in publishing in peer-reviewed papers. At CLAE we try and maintain, as do other journals, this by ensuring double blind peer review and allowing authors to select the most appropriate handling editor for their submission. Our handling editors are placed across the world (2 in Europe, 1 in the Americas, 1 in Australia and 1 in Asia) and part of their role is to encourage submissions from their region. Over the last decade we certainly have seen more and more papers from places that haven’t previously published in CLAE. In this issue of CLAE we have a true international blend of papers. We have papers from authors from the UK, USA, Iran, Jordan, France, Poland, Turkey, Nigeria, France, Spain and Brazil. I think it's a testament to the continued success of the journal that we are attracting new writers from so many parts of the world and retain papers from more established authors and research centres. We do continue to attract many weaker papers that are rejected early in the review process. Often these will be unexceptional case reports or papers describing a surgical technique. Case reports are published but only those that offer something original and especially those with interesting photographs. In this issue you will see Professor James Wolffsohn (UK) has an interesting paper around a lot of the focus of his recent research activity into clinical evaluation of methods of correcting presbyopia. In this paper he highlights predictors to aid success of presbyopic contact lenses. If you have been involved in any clinical work or research in the field of dry eye disease then you will know well the CLDEQ (Contact Lens Dry Eye Questionnaire) devised by Robin Chalmers and her colleagues (USA). This issue of CLAE details the latest research using the CLDEQ-8 (the 8 item version of the CLDEQ). The Shahroud Eye Cohort Study has produced many papers already and in this issue we see Fotouhi Akbar (Iran) looking at changes in central and peripheral corneal thickness over a five year period. These days we use a lot of new instrumentation, such as optical low-coherence reflectometry. In this issue Emre Güler (Turkey) compares that to a new optical biometry unit. Dry eye is more common and in this issue we see a study by Oluyemi Fasina (Nigeria) to investigate the disease in adults in South-West Nigeria. The TearLab™ is now commonly used to investigate osmolarity and Dorota Szczesna-Iskander (Poland) looks at measurement variability of this device. Following the theme of dry eyes and tear testing Renaud Laballe (France) looks at the use of scleral lenses as a reservoir-based ocular therapeutic system. In this issue we have a couple of papers looking at different aspects of keratoconus. Magdalena Popiela (UK) looks at demographics of older keratoconic patients in Wales, Faik Orucoglu (Turkey) reports a novel scoring system for distinguishing keratoconus from normal eyes, Gonzalo Carracedo (Spain) reports the effect of rigid gas permeable lens wear on dry eye in keratoconus and Hatice Nur Colak (Turkey) compares topographic and aberrations in keratoconus. Other interesting papers you will find are Mera Haddad (Jordan) investigates contact lens prescribing in Jordan, Camilla Fraga Amaral (Brazil) offers a report on the use of ocular prosthetics, Naveed Ahmed Khan (Malaysia) reports of the use of dimethyl sulfoxide in contact lens disinfectant and Michael Killpartrick (UK) offers a short piece with some useful advice on contamination risk factors that may occur from the posterior surface of disposable lenses. So for this issue I would say that the Gold Medal for biggest contribution in terms of papers has to go to Turkey. I could have awarded it to the UK too, but Turkey has three full papers and the UK has two plus one short communication. Turkey is also one of the countries that has shown the largest increase in submissions over the last decade. Finally, welcome aboard to our newest Editorial Board Member Nicole Carnt from Australia. Nicole has been an active researcher for many years and acted as a reviewer for CLAE many times in the past. We look forward to working with you.

Sustainable tourism practices through the eyes of the tourist: a case study of a tour operator

This internship report aim was to find out the importance of sustainable tourism practices from a tourist point of view. The research study and internship took place at Back-Roads Touring - a tour operator (based in London) that specializes in small group touring for seniors travellers from Australia, New Zealand, USA and Canada. Google Forms online questionnaire was used in order to answer the objectives of the study. In total 1322 clients received an online questionnaire out of which 182 filled in the survey. The study had four objectives, however the main focus was to find out which tourism practices ( out of 15 choices) were the most important to the respondents during a Back-Roads Tour and which tourism options (using the same 15 choices) the respondents identify as sustainable tourism options ( objective 2 and 3). The results showed that the respondents valued the most small group touring and having an experienced tour leader, which are the core practices of the company. Furthermore, the respondents were not seeking for more sustainable tourism practices neither willing to donate money to local charities/non- profit organizations. This finding shows that the clients are content with a current model of a tour, are willing to return, however price and quality will be play a key role in the decision. The respondents´ lack of wish to support or donate money to charities /non-profit organizations, could be due to the fact that most of the clients were from middle class thus they did not have enough disposable income. Moreover, one of the objectives of this study was to find out if the clients valued sustainable tourism certification. The results showed that the respondents’ were not aware of the sustainable tourism certification that the company obtained, however slightly over half of the respondents said that the knowledge of the certification has positively influenced them to travel again with a company.

Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp

High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n = 29), giardiasis (n = 47) and amoebiasis by Entamoeba histolytica (n = 3) or E. dispar (n = 10) and apparently healthy subjects (n = 24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56 °C were proven more efficient for the release of DNA from Cryptosporidium oocysts.

Percurso profissional. Destaque para a análise da produção de resíduos urbanos no Alentejo Central - portugal e utilização do modelo DEA na definição da meta da recolha seletiva

Neste trabalho de Projeto efetua-se o desenvolvimento do tema da produção de Resíduos Urbanos (RU) no Alentejo Central, utilizando-se o modelo DEA (Data Envelopment Analysis) para a análise das metas da recolha seletiva estabelecidas para 2020. Genericamente, quanto maior é o grau de desenvolvimento económico de um território, maior é a taxa de urbanização e maior é também a quantidade de resíduos urbanos produzidos por habitante. O rendimento e a urbanização são variáveis altamente correlacionadas, quando aumenta o rendimento disponível e os padrões de vida, aumenta também o consumo de bens e serviços de modo correspondente, o que leva ao aumento da quantidade de resíduos gerados. Assim, tendo em conta os impactos locais que os RU abandonados trazem, e com o objetivo de quebrar o elo entre crescimento económico e os impactos ambientais associados à produção de resíduos, são implementadas, nos países com elevados níveis de desenvolvimento, políticas baseadas em modelos integrados de gestão de RU que permitem a recuperação, reciclagem e valorização dos materiais, reservando-se a eliminação (deposição em aterro) para frações não valorizáveis, o que gera empregos e riqueza. Em Portugal vigora o Plano Estratégico para os Resíduos Urbanos (PERSU 2020) que define objetivos e metas nacionais, nomeadamente a meta da recolha seletiva, estabelecendo para 2020 um quantitativo nacional mínimo a recuperar de 47 kg por habitante por ano. Deste modo, importa caracterizar, para o período de 2002 a 2012, como evoluiu a produção de RU em comparação com a evolução do PIB em Portugal. A análise foca-se então na produção de RU na região do Alentejo em particular no Alentejo Central que evidencia um elevado nível per capita em comparação com o resto do país, situando-se mesmo acima das regiões do grande Porto e Lisboa. São apresentadas possíveis razões para o registo destes elevados níveis de produção de RU não se conseguindo, no entanto, avançar com evidências. Como o modelo DEA é utilizado no PERSU 2020 para fundamentar a projeção das metas da recolha seletiva por sistema de gestão de resíduos urbanos, fez-se a sua reprodução, o que permitiu uma análise mais detalhada dos dados e o ensaio de novos resultados considerando, para além do nível de produção de RU, o numero de equipamentos de deposição de recolha seletiva como input do modelo; Abstract: Title of the report: Professional career. Emphasis on the analysis of urban waste production in Alentejo Central - Portugal and the use of the DEA in defining the separate collection target This professional report presents the theme of the Urban Waste (UW) production in Central Alentejo, using the DEA (Data Envelopment Analysis) to analyse the target set for selective collection in 2020. Generally, the higher the degree of economic development of a region, the greater the rate of urbanization and the greater also the amount of municipal waste produced per capita. The variables income and urbanization are highly correlated, if you have an increase in the disposable income and living standards, the consumption of goods and services will increase accordingly, which leads to the increase of the amount of produced waste. Thus, taking into account the local impact that the abandoned UW brings, and in order to break the link between economic growth and the environmental impacts associated with the production of waste, countries with high levels of development implement policies based on integrated UW management models that allow the recovery, recycling and valorisation of materials, restricting the disposal (landfill) to non-recoverable fractions, which creates jobs and wealth. Portugal established a national strategic plan for Urban Waste (PERSU 2020) which defines the goals and national targets, including the selective collection target stating for 2020 a minimum recover of 47 kg per capita per year. Then it is relevant to characterize and compare the evolution of UW production and GDP in Portugal for the period 2002 to 2012. The analysis then focuses on the production of UW in Alentejo, particularly in Central Alentejo region, which shows a high per capita level compared to the rest of the country, placed just above the Greater Porto and Lisbon region. Then we explore several possible reasons for this high level of UW production in this region, but none is successful in producing strong evidence. As the DEA is used in PERSU 2020 to support the projection of the selective collection targets for the municipal waste management systems, in this report we develop the model, which allowed access to the data and a more detailed analyse. Then we introduce and test a new input, the number of separate collection deposition equipment, which gives new results that are compared with the original ones.