Assessing the impact of capacity building for HIV activists in Europe and Central Asia.

STEP-UP is a capacity building and training programme for HIV community activists in Europe and Central Asia, led by the European AIDS Treatment Group (EATG). The programme aims to empower a new cohort of HIV activists each year to engage more deeply in HIV activism at local and national levels addressing key themes such as HIV treatment literacy, prevention technologies, living with HIV, project development, clinical trials, patient participation, advocacy and policy landscapes. The expected outcome of the project is that trainees become activists with knowledge of both policy and science who advocate for change to policy and or practice. To assess the impact of the programme on individual graduates, their organisations and the communities they work with, EATG conducted an evaluation of the programme for the period of 2013-2017. The methods used during the assessment were: a desk review of information about the programme; interviews with implementers and trainees; a focus group with trainees; inputs from national and regional networks, activists and references given by trainees; and an online survey of trainees. This article evaluates the lessons learnt and forms a set of recommendations to implement in the design of future editions of STEP-UP.

Water quality status of dugouts from five districts in Northern Ghana: implications for sustainable water resources management in a water stressed tropical savannah environment.

This study was primarily aimed at investigating the physicochemical and microbial quality of water in 14 such dugouts from five districts in the northern region of Ghana. Results obtained suggest that except for colour, turbidity, total iron and manganese, many physicochemical parameters were either within or close to the World Health Organisation's acceptable limits for drinking water. Generally, colour ranged from 5 to 750 Hz (mean 175 Hz), turbidity from 0.65 to 568 nephelometric turbidity units (NTU; mean 87.9 NTU), total iron from 0.07 to 7.85 mg/L (mean 1.0 mg/L) and manganese from 0.03 to 1.59 mg/L (mean 0.50 mg/L). Coliform counts in water from all the dugouts in both wet and dry seasons were, however, above the recommended limits for drinking water. Total and faecal coliforms ranged from 125 to 68,000 colony forming units (cfu)/100 mL (mean 10,623 cfu/100 mL) and <1 to 19,000 cfu/100 mL (mean 1,310 cfu /100 mL), respectively. The poor microbial quality, as indicated by the analytically significant presence of coliform bacteria in all samples of dugout water, strongly suggests susceptibility and exposure to waterborne diseases of, and consequent health implications on, the many people who continuously patronise these vital water resources throughout the year. In particular, more proactive sustainable water management options, such as introduction to communities of simple but cost-effective purification techniques for water drawn from dugouts for drinking purposes, education and information dissemination to the water users to ensure environmentally hygienic practices around dugouts, may be needed.

Immunogenicity of the rF1+rV vaccine for plague with identification of potential immune correlates.

The rF1+rV candidate sub-unit vaccine for plague, formulated by adsorption to alhydrogel, has been demonstrated to be immunogenic in the cynomolgus macaque in a clinically relevant dose-range (5-40 microg of each sub-unit) and regimen. Following two doses of vaccine, a specific IgG titre developed in a dose-related manner with predominance of the IgG1/IgG2 isotypes. Groups of macaques receiving only a single dose of vaccine at the 40 microg dose-level had a significantly reduced peak IgG response and faster decline to baseline. Serum collected at week 5 from 19 immunised animals competed with and displaced murine Mab7.3 from binding to the V antigen in vitro. By week 53 of the schedule, although absolute IgG titres had declined, 17/19 macaque sera tested contained competing antibody, indicating the durability of a functional immune response to rF1+rV in this species. Thirteen of these week 53 sera were passively transferred into groups of naive mice, and all conferred full or partial protection against subsequent challenge of the mice with plague. Generally, those sera which were most competitive with Mab 7.3 for binding to V antigen were fully protective by passive transfer, although one week-53 serum sample was fully protective by passive transfer but not active by competitive ELISA. The early development of protective immunity in macaques was also indicated from the protection conferred on naive mice by the passive transfer of immune macaque serum collected at 2-10 weeks of the immunisation schedule. Serum samples from representative macaques within this time period also inhibited the Yersinia-mediated cytotoxicity of J774 macrophages in a qualitative in vitro assay of type three secretion.

Immunogenicity of recombinant protective antigen and efficacy against aerosol challenge with anthrax.

Immunization with a recombinant form of the protective antigen (rPA) from Bacillus anthracis has been carried out with rhesus macaques. Rhesus macaques immunized with 25 mug or more of B. subtilis-expressed rPA bound to alhydrogel had a significantly increased immunoglobulin G (IgG) response to rPA compared with macaques receiving the existing licensed vaccine from the United Kingdom (anthrax vaccine precipitated [AVP]), although the isotype profile was unchanged, with bias towards the IgG1 and IgG2 subclasses. Immune macaque sera from all immunized groups contained toxin-neutralizing antibody and recognized all the domains of PA. While the recognition of the N terminus of PA (domains 1 to 3) was predominant in macaques immunized with the existing vaccines (AVP and the U.S. vaccine anthrax vaccine adsorbed), macaques immunized with rPA recognized the N- and C-terminal domains of PA. Antiserum derived from immunized macaques protected macrophages in vitro against the cytotoxic effects of lethal toxin. Passive transfer of IgG purified from immune macaque serum into naive A/J mice conferred protection against challenge with B. anthracis in a dose-related manner. The protection conferred by passive transfer of 500 mug macaque IgG correlated significantly (P = 0.003; r = 0.4) with the titers of neutralizing antibody in donor macaques. Subsequently, a separate group of rhesus macaques immunized with 50 mug of Escherichia coli-derived rPA adsorbed to alhydrogel was fully protected against a target dose of 200 50% lethal doses of aerosolized B. anthracis. These data provide some preliminary evidence for the existence of immune correlates of protection against anthrax infection in rhesus macaques immunized with rPA.

Recorridos en el continuum de los trastornos alimentarios en una muestra de adolescentes chilenos / Routes in the continuum on eating disorders in chilean adolescent samples

El objetivo de este trabajo es describir la conducta alimentaria y características asociadas en una población de niños y adolescentes chilenos, que consultan al programa de trastornos alimentarios de la PUC.

Protective efficacy of a fully recombinant plague vaccine in the guinea pig.

A fully recombinant sub-unit vaccine comprising the protein antigens rF1 + rV has been demonstrated to protect immunised guinea pigs against exposure to 10(5) colony-forming units (CFU) of virulent Yersinia pestis. Additionally, IgG purified from rF1 + rV-immunised guinea pig serum, protected the mouse by passive immunisation against challenge with Y. pestis whereas IgG purified from the serum of guinea pigs immunised with a licensed killed whole cell (KWC) vaccine for plague, protected less well. Guinea pigs immunised with the licensed killed whole cell vaccine developed an IgG titre for fraction 1 (F1) but not for V antigen. The differential in protection conferred on the mouse by passive immunisation with guinea pig IgG, was abrogated by the use of IgG purified from guinea pigs immunised with killed whole cell vaccine supplemented with V antigen. These findings indicate that the reduced efficacy of the licensed killed whole cell vaccine formulation previously observed in the mouse can be attributed to lack of the V antigen. Cross-protection of the mouse with guinea pig IgG suggests that the recognition of neutralising epitopes in the F1 and V proteins is conserved between these two species.

Engaging with cultures Reflections on entering the ethnographic field.

This paper, by Ian Hodgson, offers a reflection on the experience of starting an ethnographic study. It provides an overview of the study, addresses key elements in entering the field and suggests questions that require further investigation, while identifying those elements that are crucial to effective observation.

HIV and combination therapy: meeting the challenge of a new era.

With the advent of combination therapy in the treatment of human immunodeficiency virus (HIV) infection, there has been a vast reduction in the number of people admitted to AIDS units as inpatients, and a marked improvement in the quality of life for people infected with HIV. Combination therapy comprises the combined administration of various powerful antiviral drugs to prevent CD4 cell destruction and minimize the chance of a rapidly replicating virus becoming resistant. This article presents an overview of the basic elements of this therapy, discussing some of the challenges that are a corollary of this new era. In particular, the implications of an extended period of survival and wellness for a group of people previously thought to be the victims of a strictly acute condition are addressed. These implications relate to therapeutic, psychological, employment, social and sexual issues. Finally, the issues facing carers (both lay and professional) are discussed.

Phenotypic changes in the lipopolysaccharide of Pseudomonas aeruginosa and Escherichia coli grown in milk-based enteral nutrition solutions.

Previous studies have shown enteral nutritional solutions (ENS) contaminated with large numbers of microorganisms from the environment or gastrointestinal (GI) tract of patients have caused respiratory infections, acute and chronic enteritis, and septicemia. The introduction of "closed" enteral feeding systems has been used to prevent contaminating organisms from entering enteral feeding systems in large numbers. However, there is some discussion as to whether this has been an effective measure in reducing ENS-related infections because there is anecdotal evidence to suggest that disease processes resulting from enteral feeding are still commonplace in the hospital and home. This is because there is very little information about the growth of microorganisms in ENS and whether growth in ENS may affect the virulence and pathogenicity of microorganisms. This study shows that Escherichia coli and Pseudomonas aeruginosa may grow at 25 degrees C from either high or low initial numbers to up to 9.2 log colony-forming units per mL in a range of milk-based ENS. However, these organisms did not grow in the fruit-based ENS. The effect on the lipopolysaccharide (LPS) of culturing E. coli and P. aeruginosa in milk-based ENS as opposed to standard laboratory media was examined using polyacrylamide gel electrophoresis. We found that there were significant qualitative changes in the phenotype of O-polysaccharide side chains of the LPS from these organisms. O-polysaccharide is known to mediate in the complement, antibiotic and bile resistance, and affect adherence. Therefore, changes in the virulence and pathogenicity of these microorganisms when cultured in ENS may be indicated. Thus, the study provides further evidence for reevaluating the microbiologic and immunologic effects of enteral feeding, especially on the microbial flora of the GI tract.

Attitudes towards people with HIV/AIDS: entropy and health care ethics.

Attitudes held by health care workers towards people with HIV and AIDS are on the whole negative, and numerous studies confirm the reality and complexity of this tendency. This paper will provide a related literature review, and identify four particular factors that go some way towards explaining the robustness of these attitudes. These are social, psychological, political and anthropological in nature. A model from classical physics will be used metaphorically to illustrate and articulate the apparent inevitability of this harmful process, and also a possible solution.

The production of PCR products with 5' single-stranded tails using primers that incorporate novel phosphoramidite intermediates.

We have prepared several novel phosphoramidites and have synthesised oligonucleotides incorporating them internally. The presence of these residues in an oligonucleotide template presents an impossible barrier to primed synthesis by Taq DNA polymerase. When extended as polymerase chain reaction products, these oligonucleotides no longer serve as templates for the polymerase beyond the insertion sites of the modified intermediates, thereby producing single-stranded tails on amplification products. These tails can then be used for solid phase capture and colorimetric detection of PCR products.

Vectorette PCR: a novel approach to genomic walking.

Vectorette PCR (or chemical genetics) is a method that enables the amplification of specific DNA fragments in situations where the sequence of only one primer is known. Thus, it extends the application of PCR to stretches of DNA where the sequence information is only available at one end. In this report, we describe the chemical genetics method and demonstrate its use in one specific application. In addition, we demonstrate how fragments generated by this procedure can be sequenced rapidly and simply using methods that have the potential for automation.

Deoxyribonucleic acid (DNA) polymorphism of the alpha 1-antitrypsin gene in chronic lung disease.

Specific gene probes were used to study restriction fragment length polymorphisms of the human alpha 1-antitrypsin gene. A polymorphism due to loss of a recognition site for the restriction enzyme Taq I was identified in eight of 42 patients with bronchiectasis and nine of 49 patients with pulmonary emphysema, none of whom had alpha 1-antitrypsin deficiency. Among a control group without lung disease the polymorphism was significantly less frequent, being found in only five of 101 apparently healthy blood donors. The deoxyribonucleic acid (DNA) polymorphism was also present in two of 14 unrelated patients with alpha 1-antitrypsin deficiency, indicating a lack of association with any specific alpha 1-antitrypsin protein phenotype. The polymorphism identified in this study may be a new marker for genetic predisposition to chronic lung disease.

DNA polymorphisms of the human alpha 1 antitrypsin gene in normal subjects and in patients with pulmonary emphysema.

Alpha 1 antitrypsin deficiency predisposes subjects to developing pulmonary emphysema and childhood liver cirrhosis. We have studied restriction fragment length polymorphisms (RFLPs) of the alpha 1 antitrypsin gene in a normal population and a group of patients with pulmonary emphysema. We have identified five RFLPs with eight restriction enzymes. The most frequent polymorphisms have been detected with the enzymes MspI, PstI, and TaqI at frequencies of 46.8%, 6.4%, and 5.0% respectively in the group of normal subjects.

Resistance to reinfection with Schistosoma haematobium in Gambian children: analysis of their immune responses.

The relationship between reinfection with Schistosoma haematobium and immunological parameters was studied in a group of Gambian children aged from 8 to 13 years. Each individual's exposure to infection was assessed from observations of water contact, cercarial densities and infected snail densities at water contact sites. Eosinophil counts were made and responses to egg antigen (SEA) and adult worm antigen (WWH) measured by ELISA. Low levels of reinfection were associated with a high eosinophil count, high levels of antibodies against WWH and SEA, increased age and low exposure. In a multiple regression analysis of the association of reinfection with eosinophil count, antibody levels, exposure, age and sex, the effects of eosinophil count and exposure were still very significant after allowing for all the other variables. The effects of the antibody levels were close to significance after allowance for exposure and eosinophil count (for WWH: P = 0.09; for SEA: P = 0.07), although the evidence was less clear after additional allowance was made for age and sex. The ability of sera from the children to recognize different parasite antigens was also examined by immunoprecipitation of labelled schistosomulum surface, WWH, SEA and S. haematobium adult worm mRNA in vitro translation products. Schistosomulum surface antigens were recognized by all the sera and there was little variation in this response. There was more variation in their responses to SEA and WWH and a marked heterogeneity in the response to in vitro translation products. However, the pattern of antigen recognition appeared unrelated to susceptibility to reinfection.