On 6 April 2009, the city of L’Aquila was hit by a violent earthquake that destroyed almost all of its medieval centre, and the surviving inhabitants were evacuated and relocated in temporary quarters or undamaged homes. The aim of this study was to investigate the perceived quality of life of the elderly population 3 years after the earthquake in relation to the social and logistic issues of new housing. The study was carried out between October 2011 and March 2012, and involved 571 subjects aged over 65 years living in the municipality of L’Aquila. The interviews took place in the surgeries of general practitioners and the city’s Department of Prevention and Vaccination in the anti-influenza immunisation period. The instrument used was a 36-item questionnaire with closed, multiple choice answers divided into the following sections: demographics, everyday activities, health and perceived health, and the quality of life in the city. The results show that, 3 years after the earthquake, the elderly population living in the new towns and temporary housing of L’Aquila have a worse perception of their quality of life than the others. They feel a certain social isolation and wish to live elsewhere. Governments faced with the problems arising from a natural calamity should take into account all of the elements making up a good quality of life and, before making choices whose impact cannot be changed, consider both their immediate and long-term social consequences. A destructive earthquake gives rise to a collective sense of mourning due to the loss of affective relations, material goods and everyday points of reference, and a consequent need to redefine living patterns. The disruption of people’s lives and their loss of certainties affect their physical and mental health in both the short term and years after the calamity –3. The lActive aging: a policy framework” (2002), their quality of life not only depends on their state of health, but also on many other variables such as their social relations, their recreational and cultural activities, and the environment. These can all play a protective role in active aging, but their absence is a risk factor for physical and cognitive decline.The elderly are among the most affected , 34, 35 It is for this reason that we decided to investigate the elderly living in the highly urbanised area of L’Aquila which, on 6 April 2009, was struck by an earthquake  that almost totally destroyed the city’s medieval centre and its immediate surroundings. After the emergency phase, some of the people whose homes had been destroyed were relocated to 19 areas with earthquake-resistant housing, which led to their decentralisation and the disruption of their social networks. They found themselves living in unfamiliar surroundings devoid of services that are still little more than dormitories almost 4 years after the earthquake. This has had a profound effect on the population as a whole, but particularly on the elderly, who are less self-sufficient in terms of mobility and, having been deprived of friendships, social gatherings and places of worship, are now isolated and unable to re-establish their everyday lives or their contacts with friends and relations. The aim of the study was to verify the impact of the change on their perceived quality of life.The study was carried out between October 2011 and March 2012, and involved 571 subjects aged over 65 years living in the municipality of L’Aquila. It investigated various factors that are important for the quality of life of the elderly  and the effect that the earthquake of 2009 has had on these variables. The interviews, which were conducted by post-graduate physicians specialising in Hygiene and Preventive Medicine, took place in the surgeries of general practitioners (GPs) and the city’s Department of Prevention and Vaccination in the anti-influenza immunisation period. The GPs invited the over-65 s in their surgeries to undergo the interviews, and all of those who agreed were consecutively enrolled. The interviews were conducted anonymously and all of the interviewees were asked to give their written informed consent after the purpose of the study had been explained to them.The standardised instrument was based on one published and validated questionnaire used within a multicentric Italian study carried on by National Research Council (CNR) , and conkappa of 0.91.Before being administered to the study sample, the questionnaire was submitted to a small group of elderly for testing-retesting, the result of which was a Cohen’s 2 or Fisher’s exact test. The continuous variables were expressed as mean values and standard deviations. The normality of data distribution was tested using the Shapiro–Wilk test. Wilcoxon–Mann–Whitney test was used to evaluate differences by gender and housing conditions on the basis of the VAS score (1–10) for each individual’s perceived quality of life. Multivariate logistic regression models were used to estimate the odds ratios (ORs) and 95 % confidence intervals of the associations between the need of a subject to leave his/her current housing conditions (outcome variable) and the relative contributions made by some demographic, living and perceived well-being variables (the explanatory variables). The propensity to socialise or not was analysed on the basis of the VAS scores of the interviewees ≥5 means the presence of perceived happiness/well-being and ≤4 its absence. The tests were two-sided and a p value of 5 % was considered significant. Spearman’s coefficient was used to measure the degree of correlation between the answers to two questions, “How is your health in general?” expressed with a score between 1 (very bad) to 5 (Very Good) and “Using a scale from 1 to 10, could you tell me how happy/content you feel in this period of your life? “.The data were analysed using the STATA/IC 12.1 statistical package.The characteristics of the study sample were analysed using descriptive statistics. The discrete and nominal variables were expressed in terms of frequency and percentages, and the differences were tested using the χThe study protocol has been submitted to the institutional review board for ethical issues approval.Sociodemographic characteristics of the sample of 571 elderly, which included more men than women (50.44 vs 49.56 %) and had a mean age of 73 years (most of the subjects were aged 65–74 years are shown in Table p = 0.013). The scores attributed to the pre-earthquake period were higher for both groups but particularly the former (7.85 vs 7.61), although the difference was not statistically significant. When asked to judge the happiness of people of the same age, the subjects living in temporary housing gave lower scores than those living in rented/owned housing , and lower than those attributed to themselves. Analysis of this perception by gender showed that the women had a more negative view of their well-being than the men both before  and after the earthquake . Although not statistically significant, the subjects’ perception of the happiness/well-being of people of the same age maintained the male/female difference, and the scores were once again lower than those attributed to themselves.Table The interviewees were then asked whether they would like to live in another area or city, and 25.09 % (n = 142) said they would. Putting this wish in relation to the subjects’ perceived well-being and housing conditions Table , our datWe then asked: “Where would you like to live?” The answer “In my previous area” was chosen by 58.61 % of the sample, whereas 2.87 % would change it, 10.66 % would move to another city, and 28.87 % had no preference. In answer to the question “With whom do you spend your free time?”, 25.09 % said with friends (mainly men: 68.53 vs 31.47 %), and only 4.56 % with their neighbours. Relationships with neighbours were declared to be formal (51.49 %), reciprocally supportive (2.11 %), conflictual (1.05 %) and good (45.34 %) (data not shown).Table p = 0.000) (data not showed).The analysis of the correlation between the perception of well-being and health status shows that an increase of the scores given to their happy/content also improves the judgment of his state of health  was to evaluate their state of well-being 3 years after the earthquake. The sample corresponded to about 4 % of the over-65 year olds living in the municipality.The analysis was based on the VAS scores of happiness/well-being attributed to themselves by the subjects as a measure of their perceived quality of life, and this measure was used to evaluate the possible variables determining their health and social welfare , particularly social activities and housing conditions as changing residence was the principal intervention after the earthquake. The relationship between the trauma and morbidity seems to be clear, and the effect of the medium-term change in housing conditions on the perception of well-being is equally important.The earthquake that struck L’Aquila in April 2009 has had considerable repercussions on the health of the people affected. Numerous studies carried out after the earthquake have revealed a 37 % increase in the prescriptions of anti-depressants and a 129 % increase in the prescriptions of antipsychotics . A studyOur data indicate that the over-65 s experience a certain social isolation: most of their free time is spent with their families (81.75 %), and their relationships with friends and neighbours are rather formal, as shown by the absence of conflicts and support. It is also necessary to bear in mind that only 1.06 % of our sample worked, which suggests that the intervewees have a lot of free time: however, this is not used for socialising as very few subjects engage in activities such as sport or recreation that would bring them into contact with their contemporaries. Only 25.09 % declared they spend their free time with friends, and no more than 4.56 % with their neighbours, a finding that is in line with the results of the 2010 survey reporting anhedonia and depression in 10 % of the interviewees .p < 0.0015) and after the earthquake (p < 0.000)  Table . This isIt is worth noting that, when the interviewees were asked to judge the happiness/well-being of their contemporaries, the scores were lower than those attributed to themselves, even when the variables of housing conditions and gender were taken into account. As pointed out in other studies , 30, 32,We can therefore state that the nexus between the change in housing conditions and the quality of life and social activities is sufficiently well documented. The creation of new residential areas in L’Aquila certainly responded to immediate need for housing, but this was not followed up by the subsequent phase of creating the services and places of socialisation that could have restored a minimum of social fabric. There has been a lack of the “weak ties” that would have allowed the elderly to confront the difficulties caused by the earthquake and their consequent relocation, and an absence of sharing the problems created by the new situation. In the case of a calamity, public assistance should take into account the particular characteristics of different age groups. As they have ceased their active role in society, the everyday life of the elderly is based on affective and relational factors, which help them to overcome their difficulties and allow them to feel themselves an integral part of the community. After the first period, during which the people of L’Aquila received incredible support from voluntary workers, the second period of relocation interrupted newly formed relationships without restoring the old, also because it was not possible for families/social groups to coordinate their destinations. L’Aquila still has 30,000 inhabitants without a permanent home and it is expected that the reconstruction will take about another 20 years. Many of our interviewees may not be able to return to their homes, and their awareness of this affects their health, mood and desire to socialise. Governments faced with the problems arising from a natural calamity should take into account all of the elements making up a good quality of life and, before making choices whose impact cannot be changed, consider both their immediate and long-term social consequences.


Based on determination of ergosterol in the air of mouldy rooms the result obtained may be compared with the culture method, due to the 1,000 times higher concentration of ergosterol in the mycelium compared with spores. However, in the analysis of outdoor air, in view of the presence of mould mainly in the form of spores and the degradation of ergosterol by UV radiation, analysis of that compound may indicate a lower level of contamination compared with the culture method.The aim of the study was to compare the content of ergosterol in different microorganisms  isolated from the air as well as in six species of moulds in their different morphological forms—live mycelium, dead mycelium, and spores. Evaluation of the level of mould contamination of the air in various places using culture method and ergosterol determination was also performed. The analysis of ergosterol was carried out by gas chromatography equipped with flame ionisation detector. For evaluation of the results, analysis of variance and multiple comparison test were used. The quantity of ergosterol in the spores of various species of mould was in the range 1.9–9.4 pg/spore. The presence of yeasts and bacteria in the air does not significantly affect ergosterol concentration, in view of the low content of that sterol in their cells (max. 0.009 pg/cell for bacteria and 0.39 pg/cell for yeast). An ergosterol concentration above 1 ng per m Ergosterol is a basic sterol forming the cellular membranes of moulds and yeasts and accounts for (depending on growth phase) 10–80 % of all sterols present in the mycelium. Hence, it is easy to detect among other cell components  and mycotoxins , spectrophotometric UV or NIR; these can be used depending on the laboratory’s equipment, although the sensitivity depends on the method of determination (the most sensitive are GC–MS–MS and HPLC: 1 ng/µl) , while the lowest correlations were obtained in comparing the method of MVOC determination (ng/h) and the culture method (R2 = 0.35), or CO2 content (m/h) and the culture method (R2 = 0.57)  or mould numbers (cfu) and the level of metabolites produced by moulds  may be low in many cases because metabolite synthesis is a strain feature, dependent to a large extent on environmental conditions, the phase of growth and the presence of coexisting microorganisms. For the above reasons, the method of ergosterol determination also takes on great practical significance.Comparative analyses of various methods for evaluating the presence of moulds in building environments and plant products have shown the greatest correlation to be between ergosterol determination and a quantitative culture method .The tests were performed in different places in each room, in six repetitions, at times of greatest activity of residents or staff, to obtain average results approximately reflecting an authentic state under the specified conditions.The microorganisms used in the study were isolated from the indoor air of residential and public buildings and of a pharmaceutical plant, and from outdoor air Table . The tes3 . The final result was taken to be the arithmetic mean of all repetitions for a given place.Mycological contamination of the air was determined using an MAS-100 Eco Air Sampler  with 100 l/min flow rate. Samples of 50 and 100 l of air were taken during 30s and 60s on an MEA medium  with chloramphenicol (0.1 %) for determination of the total number of fungi and for their isolation. Air samples were taken at three points in each place, in two repetitions. Next, the samples were incubated at 27 ± 2 °C for 5 days  with nystatin (0.2 %) and incubated at 30 ± 2 °C for 48 h, yeasts were isolated on MEA medium at 27 ± 2 °C for 72 h  and YES (Yeast Extract with Supplements), with the use of taxonomic keys  was carried out using GC apparatus . Compounds were separated using an HP-5 capillary column  of dimensions 30 m × 0.32 mm × 0.25 μm with a non-polar stationary phase in the form of 5 % phenyl–95 % methyl polysiloxane. The carrier gas used was helium, with a flow rate through the column of 2 ml/min. The temperatures of the injector (S/SL—split/splitless injector) and detector (FID—flame ionisation detector) were 250 and 280 °C, respectively. Samples were injected without split (purge activation time: 1 min). The analysis was performed in programmed temperature conditions: initial temperature 150 °C (1 min), then increased at 15 °C/min to 310 °C (5 min). Quantitative analysis was performed using an external standard  method, using Agilent ChemStation (USA) software. A calibration curve was built for ergosterol concentrations ranging between 0.35 and 3.5 µg/ml. The curve was linear in the concentration range considered R2 being 0.992.Air samples for ergosterol determination were taken by a filtration method using an AirPort MD8 sampler  with 30 l/min flow rate. Samples of volume 1000 l  during 2000s were taken on sterile gelatine filters . The analytical procedure for quantitative determination of ergosterol was based on the modified method given by Seitz et al. . ErgosteSensitivity of GC with flame ionisation detector was evaluated by calculating the values of limit of detection (LOD) and limit of quantitation (LOQ). LOD and LOQ were calculated by method based on the standard deviation of the response and the slope of the calibration curve at levels approximating the LOD ICH .The determination of ergosterol content in the cells of microorganisms was performed on prepared samples of biomass containing a known number of cells. For this purpose, biological material was taken from culture on a medium in conditions suitable for the microorganisms in question . The material was taken from three Petri dishes with media into 100 ml methanol, and the biomass was then subjected to extraction and determination of ergosterol (using the extraction and measurement methodology described above).In the case of moulds, the ergosterol content was determined in the spores, which were collected from the surface of three Petri dishes with MEA medium following culture for 7–10 days at 27 ± 2 °C (so that abundant spore formation took place). The material was collected carefully into 100 ml methanol and filtered through three layers of gauze to remove mycelium fragments. Filtration was done using vacuum pump and Buchner funnel. After each filtration, the presence of the spore fraction was checked microscopically using light microscope. The filtration was repeated to separate only spore fraction without mycelium fragments (three times). The purity and density of cells was determined microscopically in a Thoma chamber based on 20 measurements (20 samples with 10 areas of the chamber were counted).12 cfu/ml for bacteria, 3.5–4.8 × 1010 cfu/ml for yeasts, and 1.5–6.8 × 108 cfu/ml for moulds. The ergosterol content, taking into account the number of cells in the extract, was recorded in units of pg/cell  or pg/spore (moulds).The experiment was performed three times. Cells densities were in the range 1.7–3.5 × 10g. After visible hyphae were obtained, the mycelium was separated from the medium by filtration, and then, a sample of biomass of known weight was subjected to extraction using methanol and to determination of ergosterol (in the conditions described above). Additionally, a fragment of mycelium was dried to constant mass . Taking account of the dry mass content of the mycelium, the results were stated in units of mg ergosterol/g dry mass. Inactive mycelium was prepared similarly, except that after separation its growth was suppressed by pasteurisation (at 100 °C for 30 min.) and it was then stored in darkness for 2 weeks. The mycelium fraction was checked also microscopically (20 preparations in 10 samples), it was observed only mycelium fragments in the form of pellets. After this time, ergosterol extraction was performed and the dry mass of mycelium was determined.Ergosterol content was also determined in mould in different morphological forms: living mycelium, inactive mycelium, and spores (converted to dry mass). Living mycelium of particular species was obtained following 40 h of incubation in liquid Malt Extract Bouillon medium  at 27 ± 2 °C and with shaking at 150 × Material for analysis of ergosterol in the dry mass of spores was taken from ten Petri dishes with MEA medium into 100 ml distilled water, by the same technique as described above. Then, the water was evaporated until the spore dry mass was obtained. This was subjected to extraction with methanol, followed by determination of ergosterol. Results were recorded in units of ng/mg spore dry mass.All measurements were repeated in three independent experiments.p < 0.05), means were compared by the Tukey’s test at significance level 0.05.Statistical calculations were performed using STATISTICA 6.0 software . The obtained results were evaluated using one-way and two-factor analysis of variance (ANOVA) at the significance level 0.05. When statistical difference was detected (p > 0.05), which amounted to 0.008–0.009 pg/cell. However, significantly different (p < 0.001) ergosterol concentrations were found in yeasts, in which the content of that compound lay in the range 0.125–0.397 pg/cell.Analysis concerned the content of ergosterol in three groups of microorganisms: bacteria, yeasts, and moulds Table . No statp < 0.001). In statistical analysis using the Tukey’s test, at a significance level of 0.05, no statistical differences were found (p > 0.05) between the concentration of ergosterol in spores of the strain Aspergillus niger (2.125 pg/spore) and spores of the strain Aspergillus versicolor (1.923 pg/spore). The results for ergosterol concentration in the spores of the remaining four strains of mould are statistically different (p < 0.05).Based on single-factor analysis of variance, statistically significant differences were also found in the ergosterol concentration determined in mould spores (p < 0.001), while a 100 times higher ergosterol concentration was found in the cells of yeast and a 1,000 times higher concentration in mould spores.Compared ergosterol concentration between bacteria, yeast, and moulds, the lowest level is found in the cells of bacteria  of six mould strains. Analysis of variance showed significant differences Table —between The Tukey’s procedure showed that there are statistically significant differences in ergosterol concentration depending on the morphological form of mould, when the level in spores is compared with the level in living mycelium and dead mycelium, irrespective of which mould strain is tested. The spores of all tested strains of mould contained the lowest concentration of that compound (from 1.07 to 2.42 ng/mg dry weight of spores).p < 0.05) only when results are compared for the strain Cladosporium cladosporioides (9.92 mg/mg dry weight of mycelium) and for Penicillium expansum (5.93 mg/mg dry weight of mycelium).However, comparing the ergosterol concentrations in living and dead mycelium, the differences in ergosterol content are found to be statistically significant  and ranges from 3.93 to 5.40 mg/g dry mass of mycelium (Table The ergosterol content in living and dead mycelium of mould species: 3), a higher level of ergosterol was also found . In mould-infested rooms, the number of moulds reached 3.1–3.6 × 103 cfu/m3, and ergosterol content ranged from 1.4 ng/m3 (museum rooms) to 2.6 ng/m3  , where the ergosterol content reached 1.1 ng/m3. Certain species of mould were characteristic of that environment, the dominant ones being Alternaria alternata, Cladosporium sp. and Acremonium sp.The highest number of moulds was recorded in outdoor air , the ergosterol content did not exceed 1 ng/m3 (it lay in the range 0.53–0.86 ng/m3). The dominant moulds in these premises belonged to the same species: Cladosporium macrocarpum, C. herbarum, C.cladosporioides, Penicillium brevicompactum, P. chrysogenum, P. crustosum, P. carneum, Aspergillus niger, Alternaria alternata.In rooms where the number of moulds in the air was found to be at a low level, ranging from 1.6 × 10p < 0.001). The levels of ergosterol in the cells of bacteria and yeasts and in mould spores indicate that ergosterol can be used for airborne mould determination only. The concentration of ergosterol in yeast cells is ten times smaller than for moulds, and their numbers in the air do not normally exceed 101 cfu/m3 , but they are primarily linked to genus, species and strain features.Analysing the ergosterol content in spores as obtained in the present work and in earlier studies . Earlier research by Robine et al. , Penicillium expansum (7.4 pg/spore), P.chrysogenum (5 pg/spore). In the case of airborne moulds of the genus Aspergillus or Alternaria, the ergosterol may prove difficult to detect. However, studies have shown large variety of species among the moulds found in air, which provides an argument for the possibility of estimating mould numbers based on ergosterol measurements. Another such argument is the presence of mould in the air in the form of both spores and mycelium fragments, as the ergosterol concentration has been shown to be 1,000 times higher in mycelium than in spores. Previous research has shown that mycelium fragments make a significant quantitative contribution to the pool of all forms present in the air and also have more powerful allergic effects than airborne spores  were found between the concentration of ergosterol in living and non-living mycelium, except of mould species Cladosporium cladosporioides and Penicillium expansum. Similar conclusions were reached by Mille-Lindblom et al. , there was observed to be a lower concentration of ergosterol (1.0 ng/m3) compared with highly mould-infested rooms (3.1–3.6 × 103 cfu/m3), which had an ergosterol content of 1.4–2.6 ng/m3.Mille-Lindblom et al.  also not3). In rooms in which there was active development of moulds on building partitions or other objects, a significant airborne contribution may come from fragments of living mycelium, in which case the ergosterol concentration may be high and overestimated compared with the results obtained by the culture method. This situation was found in residential buildings where mould was observed on the walls, and in museum storerooms containing mould-infested historical objects. The highest level of ergosterol concentration in the air was found in residential premises; in 10 buildings it ranged from 1.4 to 6.4 ng/m3 air.The ergosterol content in the spores is low, as was shown in our work, probably due to the limited number of mitochondria in spores . Future studies should also include a detailed analysis of taxonomic units and morphological forms in the investigated environments.Based on comparison of the results of analysis of ergosterol and of moulds numbers determined by the culture method, it can be assumed that a concentration of ergosterol above 1 ng/mThe presented study confirms the possibility of using ergosterol as the estimated indicator of mould’s presence in the air in the form of spores and fragments of live and inactive mycelium. However, it should be considered, that, depending on the morphological form of moulds presented in the air and the kind of environment (indoor and outdoor air) indicator may show the different result than obtained in culture method. Therefore, the determination of the airborne moulds amount by multiple methods should be performed, which would provide various information such as the number of living cells and the total microbial biomass in the air. An important issue is the development and validation of the air sampling methods for microbial contamination analysis, which are the first and key steps of the environment assay.


Musa paradisiaca (BanLec-1) could inhibit the cellular entry of human immunodeficiency virus (HIV). In order to evaluate its effects on tobacco mosaic virus (TMV), the banlec-1 gene was cloned and transformed into Escherichia coli and tobacco, respectively. Recombinant BanLec-1 showed metal ions dependence, and higher thermal and pH stability. Overexpression of banlec-1 in tobacco resulted in decreased leaf size, and higher resistance to TMV infection, which includes reduced TMV cellular entry, more stable chlorophyll contents, and enhanced antioxidant enzymes. BanLec-1 was found to bind directly to the TMV capsid protein in vitro, and to inhibit TMV infection in a dose-dependent manner. In contrast to limited prevention in vivo, purified rBanLec-1 exhibited more significant effects on TMV infection in vitro. Taken together, our study indicated that BanLec-1 could prevent TMV infection in tobacco, probably through the interaction between BanLec-1 and TMV capsid protein.It has been demonstrated that the lectin from  A wide array of abiotic and biotic stresses seriously affects plant growth and crop yield. Among the various biotic stresses, viral diseases represent one of the most significant reductions to crop quality and productivity. When expLectins are a huge protein family that recognize and bind to specific carbohydrates, and function as central mediators of information transfer in biological systems by interacting with glycoproteins, glycolipids and oligosaccharides. Since anBanLec-1, a glucose/mannose-specific lectin, was isolated from banana and partially characterized. It is a BanLec-1 has been recognized as a potential immunomodulatory molecule. Swanson Musa paradisiaca) fruits were purchased from the local market. Total RNA was isolated by using Trizol reagent . The cDNA sequence was amplified by Reverse transcription polymerase chain reaction (RT-PCR). Primers for banlec-1 cDNA (GI:37223481) were 5′ATGAACGGAGCGATCAAGGT and 5′TTATGGCTCCAAGTAGACCC. The amplified product was gel purified and cloned into binary vector pCAMBIA1304.1, which was then transformed into Agrobacterium tumefaciens strain GV3101. Leaf discs of Nicotiana tabacum NC89 were transformed. Transformed plants were selected on Murashige and Skoog medium (MS)  × 100), and disease index (%) was calculated as {∑[Disease progression × numbers of plants of corresponding disease progression] / [The highest disease progression×number of tested plants]}×100).TMV was prepared freshly by grinding 0.5 g of young systemically TMV-infected tobacco leaves in 2 mL of ice-cold 10 mmol/L PBS, pH 7.4. The resulting suspension was centrifuged at 100 g for 5 min at 4 °C. For To measure the chlorophyll content, 0.5 g of tobacco leaf filaments were soaked in 5 mL of 80% (v/v) acetone until the colour of the filaments completely changed from green to white. The supernatant of the extract was re-set to 5 mL and taken for light absorption measurement at 663 nm and 645 nm, respectively.4 for 2–3 h. After fixation and dehydration by ethanol, leaf samples were washed by acetone for three times, and embedded in Spurr's resin. Ultrathin sections were sequentially stained with uranyl acetate and lead citrate, followed by examination under a transmission electron microscope.Rectangular segments (1.5 mm × 1 mm) were cut from the leaves of transgenic and wild-type (WT) lines between the third and the fourth principal vein. The segments were fixed for 2 h in 2% glutaraldehyde, then washed in PBS buffer three times, and fixed again in 1%–2% OsOg for 10 min at 4 °C. The supernatants were used for determination of superoxide dismutase (SOD) and peroxidase (POD) activities.[Leaves of WT and transgenic lines were grinded in ice cold 50 mmol/L PBS (pH 7.8) containing 0.5% Triton X-100, 10 mmol/L β-mercaptoethanol, 0.1 mg/mL serine proteinase inhibitor (PMSF). Crude tissue homogenates were centrifuged at 12,000 ×tivities.Purified rBanLec-1 and whole cell extracts after IPTG induction were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and electro-transferred onto polyvinylidene difluoride (PVDF) membrane. The membrane was then washed for 15 min in buffer A . The washed membrane was denatured for 2 h at room temperature in denaturation buffer , 50 mmol/L dithiothreitol (DTT), 50 mmol/L Tris–HCl, pH 8.3). After a brief wash in tris-buffered saline  and renaturation buffer , 0.1% Tween-20, 2 mmol/L DTT), the blotted proteins were renatured in renaturation buffer overnight at 4 °C, and then the PVDF membrane was blocked with 1% BSA in renaturation buffer for 90 min at room temperature. For TMV capsid protein (CP) binding, the membrane was incubated for 90 min at room temperature with 10 μg/mL of TMV CP in the renaturation buffer. After incubation, the membrane was washed four times with TBST (TBS plus 0.1% Tween-20) and incubated for 1h at room temperature with rabbit anti-TMV CP antiserum (1:1000 dilution) for 1 h. After four washes with TBST, the membrane was detected with horseradish POD and immunoglobulin G (HRP-IgG).banlec-1 gene (GI:37223481) was cloned and the resulting protein with His6-tag was expressed in E. coli C43 under IPTG induction. After purification through high-affinity Ni-charged resin, the purified product was analysed by 12% SDS-PAGE and the appearance of an approximately 15 kDa band in the gel indicated the purity of rBanLec-1 after 3 h of IPTG induction (Tm) of 60.8 °C.[2+, Mg2+ and Mn2+ were found to be able to support agglutination, whereas Zn2+ was not . In contrast, pre-incubation in vitro effectively inhibited TMV infection in a dose-dependent manner, and 12.9 μg/mL of rBanLec-1 inhibited TMV infection completely. Gel overlay assay indicated that IPTG induced whole cell extracts and purified rBanLec-1 at different concentrations could bind specifically with TMV CP. The horse radish peroxidase (HRP) signal bands exhibited at about 15 KDa further demonstrated that BanLec-1 could interact with TMV CP directly (It has been reported that BanLec-1 could bind with human immunodeficiency virus (HIV) coat protein gp120 and prevent HIV infection. At the sdirectly .Figure in vivo prevention was limited, i.e. once TMV replicates itself in cells and its content exceeds the effective concentration of BanLec-1, infection still occurs. That is why the disease rate remained at zero in transgenic plants in the first 6 d after infection, and reached 100% after 18 d , Jiangsu, China.


Mentorship programs have been found to be of great use in all career fields, especially medicine. Given that the practice of medicine is best learnt by a hands-on training, a mentorship program can prove invaluable for the young clinician scientist. A structured super-speciality training program can help in both, acquisition of clinical and research skills, as well as career growth.How to cite this article: Bhartiya S, Ichhpujani P. How to Choose a Mentor? J Current Glau Prac 2013;7(3):128-129. The word ‘Mentor' has its roots in Greek mythology. Mentor was the son of Heracles and Casopis. In his old age Odysseus placed mentor in charge of his son Telemachus, when he left for the Trojan War. When Athena, the goddess of heroic endeavor visited Telemachus she took the disguise of mentor to hide herself from the suitors of Telemachus' mother, Penelope. As mentor, the goddess encouraged Telemachus to stand up against the suitors and to find out what happened to his father. Because of mentor's relationship with Telemachus, and the disguised Athena's guidance for dealing with dilemmas, the name ‘mentor' has been adopted in English as a term referring to a person who imparts knowledge and shares his/her experience with someone less experienced.Plutarch has rightly said,‘The mind is not a vessel to be flled, but a fre to be kindled.'A good mentor helps kindle this fre, helps one set a new career path, or find greater fulfllment in their chosen direction. In the present era, a mentor can also help network, and introduce his/her protege to opportunities and people that he/she would not otherwise have access to. A mentor provides guidance and inspiration, and can be a highly credible reference for grants, awards and professional positions.15 and has been found to be imperative for facilitating a young medical professional's career advancement and acquisition of clinical and research skills.515Mentoring is increasingly viewed as a key factor contributing to a successful career in all fields including academic medicine,When you decide to go in for a mentorship program it is important that you are clear about what it is that you want in your mentor, and what kind of help and advice you are seeking. These guidelines will at best help you narrow down your choice.13It is essential to choose someone you can respect, and of course, that this person has the necessary expertize in your specific field. It also helps if you admire this particular person for his or achievements, because very often, you will have to follow your mentor's advice against your better judgment.Also, speak to past mentees, and how they found their experience with the particular person. In case you are not ready to become a clone, look for a mentor who will direct you toward your goals.You do not need to find someone who has achieved the most in his or her field: just someone who has enough generosity of spirit to let you learn from their past experiences. You will, however, find that most successful people have this quality and are secure enough to give you credit whenever its due. Again, speaking to those who have been mentored will provide you enough insight to arrive at an informed decision.It is essential that your mentor be accessible. In case a one-on-one interaction is not possible, someone who is accessible by email is good enough. In case you are looking at a surgical grooming, it makes sense to find a center which has a good wetlab and archives of training videos.A very busy and important teacher may be able to give you tips in an intensive, short mentorship program. This short burst of teaching requires you to be read and receptive at that particular point in time. But, in case you are looking for a holistic, more thorough training program, your best bet would be a teaching hospital with a structured training program. Whereby, several teachers can ensure that your learning is comprehensive and complete.Very often, a mentorship program headed by a truly brilliant clinician-teacher, supported by an able mid-level supporting crew is best suited for most of our needs. Where the humdrum and the routine is learned by passive and active imbibition of clinical practices and the truly inspirational quality is attained by association with the head mentor.For many of us, lack of financial support is a major deterrent when choosing a training program. A mentor who can provide part or complete financial aid to support the training is an obvious choice in this case. It is also judicious to apply for local and national societies for help; most offer small to medium term grants to help doctors in training.Keeping your personal goals in mind, you can evaluate the productivity of the mentorship program you are applying for. In case your aim is to be involved in research and publications, evaluating the projects which the previous mentees have worked on is an easy process. Also, the number of publications from the proposed mentor's team in the last 5 years is an indication of the group's research interests.If your interest is primarily surgical training, then it is imperative to associate with a high volume practice. Surgeries that percolate down to a surgeon in training are a fraction of the total pool, therefore, it is logical that a busy surgeon will be the best suited for your needs.Get out of your comfort zone. It is a cultural as well as an academic exercise.A good mentor can help you identify and actualize your career goals. A capable mentor will also enable you target weaknesses, build on leadership skills, and help formulate your career plan. It is therefore, a decision that must be taken judiciously.To sum it up, look for a ‘Mentor' whose hindsight can become your foresight.


Eating disorder (ED) admissions to the Paediatric Inpatient Unit at Flinders Medical Centre increased sharply between 2008 and 2012. In response, the Paediatric Eating Disorder Program was established in February 2013. The multi-disciplinary program is based on the principles of the Maudsley approach, and consists of a 4-level system including medical stabilization, establishing normal eating and transition to home. To evaluate patient outcomes in the new program, a research project commenced in August 2013. In the six months to February 2014, 25 eligible patients were assessed. This patient group displayed severe ED psychopathology at a level consistent with clinical ED populations and significantly higher than matched non-clinical norms. Mean BMI at admission was 16.38 (SD = 2.05). After an average length of stay of 20.15 days (SD = 12.78), the mean discharge BMI was 18.19 (SD = 1.76). The number of readmissions has decreased since the program started: 39% of patients had more than one admission in 2013 compared to 47% in 2012. We present a sample case to highlight the complexities of this patient group, and to provide a practical description of the treatment approaches employed in the program.Service Initiatives: Child and Adolescent Refeeding and FBT stream of the 2014 ANZAED Conference.This abstract was presented in the


Nursing education has evolved over time to fit societies’ increasing care needs. Innovations in nursing education draw thorny debates on potential jeopardy in the quality, safety, and efficacy of nurse graduates. Accelerated nursing education programs have been among landmark strategic changes to address the persistent bedside nurse shortage. Despite the dearth of empirical studies in sub-Saharan Africa (SSA), the National School of Public Health of Burkina Faso has developed a State Diploma Nursing (SDN) fast-track program. With innovative features, the program is nested into the traditional SDN program. This study investigates preliminary outcomes of the implemented policy using the initial cohort that went through the program. Comparison of the traditional generic program and the fast-track one is drawn to inform nursing education policy.t-test, F-test, and multivariate ordinary least square regression (OLSR) were employed to determine the comparative pattern between the traditional generic and the newly nested fast-track program. Students’ varied statuses  were kept to better outline the findings trend.The study was conducted in the three campuses delivering the SDN program. Data collected from a representative sample included 255 students from the 2006–2009 cohort, after concluding the program. Surveyed students were assessed according to the program entry status. Outcomes were measured using students’ academic performance. Besides descriptive analysis, bivariate p < 0.001), and further informed differences between campuses. Students entry status also displayed significant differences, yet the academic performance of post-registered students from traditional generic versus fast-track was similar (p = 0.409).A fifth (19.6 %) of surveyed students were enrolled in the fast-track stream from which, one third (33.7 %) consisted of post-registered students. Fast-track students comparatively achieved the best academic performance . Multivariate OLSR confirmed that fast-track students performed better (β: 5.559, Findings suggest that fast-track program students performed better than the ones from the traditional generic program. The uniqueness and success of this mixed nursing program experience sheds light for nursing educators engaged in policy making. The study results can serve as a crucial foundation for policymakers to alleviate the nurse shortage in SSA. Nursing education has demonstrated a phenomenal growth in term of program numbers, variety or content delivered . DespiteThis concern affects the Masters of Science in Nursing (MSN) programs and especially the Bachelor of Science in Nursing (BSN) . The litThere is an established shortage of adequately trained professionals worldwide, including in resource-constrained settings like Burkina Faso . This shThe concept of fast-track program dates back to the 1950s, when an ADN education program was developed to meet short-term system needs of the post World War II United States (US) . It was Studies comparing the traditional generic students’ performance to accelerated ones showed similar to better performance of the latter group. Some studies, however, did not find differences between fast-trackers and traditional generic students’ grades . NeverthFather Goarnisson pioneered nursing training in Upper Volta (current Burkina Faso). The first formal nursing school, the National School of Nursing, was founded in 1968  and upgrNon-nurse candidates looking for government position may sit for a national test. Successful candidates attend the SDN program as state pre-registered students, granted a stipend, waived from paying tuition, and serve the government upon completion. Candidates applying privately are given the status of private pre-registered students.State ADN may enter the SDN program after being shortlisted in the national test. Five years of experience as ADN is a pre-requisite. They are labelled state post-registered students. Under the new scheme, students attend the fast-track program for two calendar years and because of the nested feature, they directly join the SDN program in their sophomore year.Non-government ADN may apply for the SDN program as private post-registered students. So far, there is a vacuum in the selection policy; their recruitment to the program is based on merit.As elsewhere , ADN proStudents sit for a summative examination at the end of both year 1 and year 2. The third year summative examination serves as a graduation examination. This arrangement is similar to what is done in Norway and in Australia  and in sThe innovative fast-track in Burkina Faso nursing education is unique. The current program contains features that are not common in the literature. In the nested scheme, students attend the same classes, practicum, and rural setting internship in the third year. The fast-trackers are not required to take any transitional courses but must pass a paper-pencil test. ADN to SDN fast-track program prevents useless repetition  as studeWhile accelerated programs have proven to be at least as effective as the traditional generic programs in industrialized countries, very little is known about the implementation of such initiative in SSA such as the fast-track SDN program implemented in Burkina Faso. This study intended to achieve the following objectives: 1) to evaluate the performance of fast-track students of the 2006–2009 cohort, 2) to compare the performance of students in the traditional generic program to those in the fast-track SDN program, and 3) to draw a conclusion on whether the fast-track policy is justified.The study was carried out at NSPH, which is a government-owned institution with six outlying campuses. The three campuses that offer the SDN program were included: Ouagadougou, the main campus and two outlying campuses, Bobo Dioulasso and Ouahigouya. We conducted a quasi-experimental survey design with a descriptive and comparative approach. Participation comprised of 255 students who were blindly captured from a total of 356 who formed the 2006–09 cohort. This cohort was the first to include a fast-track program. Only three students failed and were excluded from participation. Of these 255 students, 50 attended the fast-track program (2-year SDN program) while 205 were registered in the traditional generic program (3-year SDN program). Among the latter, 36 were post-registered (last batch of post-registered enrolled in 3-year SDN program).All students, from both the fast-track and generic programs, were invited to take part in this study through an announcement posted at the school. Students who chose to participate were informed in detail on the nature of the study, their rights and responsibilities, and were asked to sign a participation agreement form. Data were collected in two phases. On phase one, students were asked to fill out a self-administrated questionnaire. Demographic, socio-economic and achievements data were gathered from both generic and fast-trackers. For phase two, the Principal Investigator (PI) (BI), conducted a document review from the campuses’ academic affairs, to solicit all students’ academic scores (course-work and year-end exam scores). In order to protect participants, no identifier was assigned. Collected data were restricted to information required for performance analysis for the purpose of the study.The instrument employed was derived from the Clinical Nursing Competence Questionnaire . It is aThe questionaire has undergone a content validation, forward from English to French and backward as translated by a professional translator. Five experienced English teachers from the English department of the University of Ouagadougou in Burkina Faso, worked independently, examining every step of the translation process.The pilot test was conducted between June 16th and 30th, 2009 and included 10 nursing students from Bobo Dioulasso, and 10 others from the Ouagadougou campus, using a convenient sampling. Participants were asked to complete the form impromptu. The response rate was 100 %. Minor adjustments occurred for demographic items such as the structure of the birth data, some items were reworded. The pilot test was run to ensure better ways of presenting the tool items for more accurate responses and ratings.The dependent variable consisted of students’ academic performance stratified by track. The explanatory variables were categorized into demographic and academic variables. As indicated in Table The study was first authorized by the National Taipei University of Nursing and Health Sciences Institutional Review Board . Furthermore, we obtained authorization from the NSPH headquarters and the each campus principal, so that the academic affairs department from campuses could allow data access to the PI for on-site use. The participation was completely voluntary, anonymous, and freedom to withdraw in the process was guaranteed. Before the survey questionnaire was administrated, every participant provided signed written informed consent.t-test and ANOVA. Finally, a multivariate analysis was conducted using ordinary least square regression (OLSR). We assumed equivalence among students of post-registered status. Therefore, to figure out the true effect of track, we excluded the post-registered students from the generic program from the multivariate analysis. They could have been fast-trackers if the policy was implemented a year earlier, or if they had entered a year later. Data were entered without personal identifiers and analyzed with SPSS 18 for Windows. The two-tailed was tested with 95 % confidence level  considered statistically significant.This survey employed a three-stage analytic design. First, participants were described by track. The measures of central tendency were computed for each group. Next, group comparisons were examined using 2-sample n = 50) attended the fast-track, 42 were post-registered students attending the 3-year SDN program (the last group). Two-third were male, while half of the students were under the age of 30 (n = 129). The majority (58 %) of students were from the main campus (Ouagadougou) (Table t-test shows an average of 7.5 more years for the latter group (p < 0.001) (not shown). Elsewhere, accelerated program students tend to be the oldest [This study on the fast-track program nested in the traditional generic SDN program is a baseline assessment of a project aiming to provide evidence concerning the policy’s worth. This assessment targeted the first cohort 2006–2009) that was unique in many ways. First, it includes both the last batch of post-registered students who sat for the 3-traditional generic SDN program and the first intake of fast-trackers. The two groups are used as proxies of the former system and the new policy implemented, respectively. Assessment of this group allowed a critical analysis between comparable subsamples. Second, all three campuses offering the program at NSPH were committed to its implementation. Finally, this initial assessment served to evaluate the policy in order to inform future developments of the program. Of the 255 students participating, one out of 5 (006–2009 f 30 n = 29. The me oldest . Unsurprp < 0.001). Between tracks differences show that fast-trackers have 5 points more than traditional generic students .For comparability between tracks, analyses included students’ performance achieved from sophomore year. Table In the regression model, gender, marital status, and number of children did not reveal any statistical association. Because of family strain impact, this absence of difference to a certain extent violates the common perception that accelerated programs are notoriously intense, therefore demanding. Earlier studies pinpointed challenges like family, friends, and leisure foregoing  as affecr: 0.844, p < 0.001), underpinning a high collineality. We therefore ran two models. The first integrated registration status to distinguish between pre-registered (state and private). The fast-trackers, on average, have 4.5 points more than generic state registered students . In model 2, fast-trackers performed significantly better than generic track students . This finding confirms evidence from earlier studies on pure performance [The assessment of track and registration status into the model led to a higher correlation . The comparison of their annual score suggested a similar trend  also performed significantly better than those who were pre-registered (State and private) Table . This hiThis mixed program is unique in literature and the present work is a baseline assessment of the innovative nested fast-track SDN program implemented in Burkina Faso. It compared the performance of students enrolled in nested fast-track programs introduced by the NSPH, being the first of its kind in nursing education literature. As the baseline assessment, this single cohort-focus study has the unique merit of gathering the initial fast-track batch, and the last group of post-registered attending the 3-year traditional generic program. Our findings suggest the performance standing for the upcoming cohorts, provide the basis for future studies, and finally inform appropriate policy options. We can acknowledge the plausibility of findings as participants took the same courses within the same educational context and conditions for comparability. A significant difference between tracks and the similarities among post-registered students confirmed that fast-trackers’ academic performance was considerably better, justifying the worth of its implementation. This unique, but progressive example may inspire other nursing schools, especially from SSA to reform their programs to suit their contextual needs and resources.Findings show significant differences between the campuses in the same school, raising critical questions in the program delivery, student prerequisites, and co-requisites. From these important differences within the same institution, we can assume that the discrepancy may exist and may even be huge with other nursing schools in the country . The NSPH has more than 40 years of history and is thus experienced in nursing education, administration, and clinical placement networks. As students’ educational quality declines  in the aThis study had some strengths and limitations. First, study participants shared the same learning environment, which made it possible to control most external factors that could have influenced their performance. Second, the sample size was large enough and included all registration statuses codified in the country. Though our study used grades, including clinics, some criticized a certain reductionism when approaching clinical nursing competence . FinallyRegistration status: academic designation defining the level or position or prior background of candidate-students while she/he is being enrolled in the nursing program.Pre-registered: are novice students entering into the SDN program without any prior background in nursing.Post-registered: are registered ADN re-entering into the nursing program to equip themselves with further specialized practice and knowledge. In Burkina Faso, those candidates must hold 3 to 5 years of bed-side experience.State registered students: are students recruited into the program through the mechanism of a national test. They are provided a stipend, exempted from paying any charge related to registration, tuition and are systematically employed by the government on completion.Private registered students: they are either novice (pre-registered) or re-entering nursing education (post-registered) and required to pay tuition fees.Academic performance: is the measurable outcome of scholastic achievement, shown through test scores and total course points earned.Fast-track program: Accelerated nursing program taken for two calendar years upon an ADN program completion.Traditional generic program: is the traditional 3-year nursing program attended by new non-nurse applicants.


Violence against adolescent girls in humanitarian settings is of urgent concern given their additional vulnerabilities to violence and unique health and well-being needs that have largely been overlooked by the humanitarian community. In order to understand what works to prevent violence against adolescent girls, a multi-component curriculum-based safe spaces program  will be implemented and evaluated. The objectives of this multi-country study are to understand the feasibility, acceptability and effectiveness of COMPASS programming to prevent violence against adolescent girls in diverse humanitarian settings.Two wait-listed cluster-randomized controlled trials are being implemented in conflict-affected communities in eastern Democratic Republic of Congo (N = 886 girls aged 10–14 years) and in refugee camps in western Ethiopia (N = 919 girls aged 13–19 years). The intervention consists of structured facilitated sessions delivered in safe spaces by young female mentors, caregiver discussion groups, capacity-building activities with service providers, and community engagement. In Ethiopia, the research centers on the overall impact of COMPASS compared to a wait-list group. In DRC, the research objective is to understand the incremental effectiveness of the caregiver component in addition to the other COMPASS activities as compared to a wait-list group. The primary outcome is change in sexual violence. Secondary outcomes include decreased physical and emotional abuse, reduced early marriage, improved gender norms, and positive interpersonal relationships, among others. Qualitative methodologies seek to understand girls’ perceptions of safety within their communities, key challenges they face, and to identify potential pathways of change.These trials will add much needed evidence for the humanitarian community to meet the unique needs of adolescent girls and to promote their safety and well-being, as well as contributing to how multi-component empowerment programming for adolescent girls could be adapted across humanitarian settings.NCT02384642 (Registered: 2/24/15) & NCT02506543 (Registered: 7/19/15).Clinical Trials  Globally, approximately 59.5 million people were forcibly displaced in 2014 , 7.Within humanitarian emergencies, female adolescents aged 10–19 years are particularly at risk of sexual violence, sexual exploitation, and early marriage , 3, 8. FNot only do adolescent girls face increased vulnerability to violence, similar to development contexts, the needs of adolescent girls are often overlooked by communities, families, and service providers during and after conflict due to their lack of social status, stemming from gender inequalities and the invisibility of younger populations. Girls may also have limited mobility and therefore, face restricted access to pertinent life-saving information, schooling, and health and protection services , 14. LimDespite the multitude of reports on violence against women and girls in humanitarian settings and the damaging physical and mental health effects resulting from such violence, few rigorous studies have examined how best to prevent violence in such contexts , 18. In To address this gap in the evidence, Columbia University and the International Rescue Committee (IRC), are undertaking an evaluation of the Creating Opportunities through Mentoring, Parental involvement, and Safe Spaces (COMPASS) program, of which a key strategy is to develop the skills and assets of girls with the support of female mentors in ‘safe spaces’, alongside additional engagement of caregivers and capacity-building of service providers. The objectives of this multi-country study are to understand the feasibility, acceptability and effectiveness of these approaches to respond to and prevent violence against adolescent girls in three diverse humanitarian settings.Over three years 2014–2017), the COMPASS program is being implemented and evaluated in conflict-affected communities in the Democratic Republic of Congo (DRC), Sudanese refugee camps in Western Ethiopia, and in sites across Khyber Pakhtunkwa province in Pakistan, including Jalozai camp. In DRC and Ethiopia, the study design employs a wait-listed two-arm cluster randomized controlled trial. Given the exceedingly restrictive environment in Pakistan around issues of violence against girls, a mixed methods pre-post test is being implemented rather than an impact evaluation. Thus, the remainder of the study protocol manuscript will center on the trials taking place in DRC and Ethiopia. Further details of all study designs are found in Table 4–2017, tEastern DRC has grappled with protracted conflict for decades, vacillating between periods of relative calm and instability. North and South Kivu provinces in eastern DRC are home to approximately 60 % of DRC’s 2.6 million internally displaced persons as well as continual influxes of refugees from Burundi, Rwanda, or other neighboring countries . ViolencThree refugee camps in the Benishangul-Gumuz region of Ethiopia, bordering Sudan and South Sudan, serve as the second study site. Approximately 38,000 people reside in these three primarily Sudanese refugee camps , which are characterized by multiple safety and protection concerns. Documented in the IRC Community Wellbeing Initiative (CWI) programming assessments, concerns include high risks of adolescent girls experiencing sexual violence prior to arrival in Ethiopia and early or forced marriage. Often, these marriages are seen as a mechanism to ‘protect’ girls from sexual violence or as a means for girls’ to meet their basic needs . These ctukuls  in Ethiopian refugee camps for only women and girls, and dedicated spaces in community based women’s organizations in DRC. These safe spaces were developed specifically for girls through several community and caregiver consultations.The theory of change for the COMPASS program is presented in Fig. Caregivers will also participate in monthly discussion groups which seek to increase knowledge of the needs of adolescent girls. The goals are to ultimately shift gender-inequitable attitudes about girls in the home and increase support to keep girls safe within and outside of the home. This content is delivered by IRC program staff. The incremental effectiveness of this caregiver component is being tested in DRC, while in Ethiopia we are evaluating the overall COMPASS program effectiveness.In addition, COMPASS will engage health service providers and case managers to train workers on Clinical Care for Sexual Assault Survivors  and CariBased on the assumptions of 80 % power to detect a Cohen’s effect size of d = 0.3 between treatment versus wait-list groups , a sample size of 704 girls is required in each country. After accounting for an expected attrition rate of 25 % in DRC, the final sample size required at least 880 girls and a total of 886 girls were surveyed at baseline. Primary caregivers of selected girls in DRC were also asked to participate in the study (n = 777 surveyed at baseline). In Ethiopia, given movement concerns within the refugee camps and higher likelihood of loss to follow-up and potential refusals at baseline, the attrition rate was inflated and a total of 919 girls were surveyed at baseline.Groups are comprised of 20–25 girls each, based on geographic proximity or language. Groups of girls were divided into younger groups and older groups, given age-tailored content of programming. In DRC, 35 girl groups across 14 villages in South Kivu were divided into two age strata: 10–12 years of age and 13–14 years of age. In Ethiopia, 47 groups across three refugee camps were divided into age strata; group formation also accounted for school scheduling in the camps. Groups were first stratified by age level (13–14 years and 15–19 years) and then randomized to either the intervention or wait-list arm following the baseline survey. If sisters or girls who identified the same caregiver or reside in the same house were participating in the program, girls were assigned to the same group if they were in the same age category. If they were in separate age groups, the groups were paired together and randomized to the same study arm. Randomization was completed using the statistical software R version 3.2.1.IRC program staff introduced the COMPASS program to the communities, and interested girls and their caregivers were invited to participate in the program. For example, in Ethiopia, outreach efforts included social worker house visits and ongoing activities at IRC Women and Girls Wellness Centers, which provide basic socioeconomic empowerment programming for women and girls in the camps and psychosocial services for survivors. Upon completion of program registration forms, girls (and their caregivers in DRC) were invited to participate in the studies. Eligibility criteria for the program and study varied by age and language based on contextual tailoring of adolescent girl programming across country sites. Details are provided in Table Throughout the delivery of COMPASS, girls and their caregivers will be tracked by program mentors to improve and record retention through attendance records. Primary concerns for loss to follow-up for this study includes displacement, lack of buy-in from parents which could result in girls not being allowed to participate, and early marriage. Specifically, the latter is a concern because when girls marry, they may drop out of programming to undertake additional household duties. If a girl becomes married within the duration of the program, she will still be encouraged to participate by the mentors. In addition, if a girl has not attended programming, mentors will directly follow-up with the girl and caregiver and support her attendance. Reasons for non-attendance will be continually assessed by program and research teams; program delivery timing or other barriers to attendance will be addressed as possible. If there are multiple girls that miss more than three sessions within one group, IRC staff will engage directly with the mentor to support that mentor to improve retention of girls in her group.Further, in Ethiopia, additional steps are planned to retain and track girls in the wait-list group, while minimizing risk of contamination. This tracking is also needed to determine whether girls have moved from the camps during the study period. These steps include checking in with girls during standard school-supply distributions approximately six months into the study and an additional check in with girls one month before endline data collection.All study procedures were approved by the Columbia University Institutional Review Board (IRB) and by in-country local bodies . The study protocol for Ethiopia was also approved by the International Rescue Committee internal review board. All caregivers were asked to provide informed consent for the girls’ participation in the study if the girl was under 18 years old. Subsequently, girls were asked to assent for their participation in the study. In DRC, informed consent was read to potential participants through trained enumerators and written consent was given. Caregivers were also asked to provide written informed consent for their own participation in the quantitative survey and qualitative research activities. In Ethiopia, informed consent was administered via audio recordings because the languages selected for the study are non-written; thus, informed consent forms were verbally translated into the appropriate languages and recorded. All potential participants listened to the same audio recording to ensure consistency in the informed consenting process and provided verbal consent. Data collection staff were trained and available to respond to any questions on the consenting process. Since languages were non-written, only verbal consent/assent was required for participants in Ethiopia.Given the highly sensitive nature of conducting research on the topic of violence against girls, all research staff completed training on the importance of ensuring confidentiality, privacy, and basic concepts of gender-based violence in accordance with the World Health Organization guidelines on violence against women research  and protThe primary outcome measure for the study is change in sexual violence. Other secondary outcomes include physical/emotional abuse, healthy relationships, and self-esteem, for example. Hypothesized directions of changes, primary and secondary outcomes, and measures are found in Table In DRC, Audio Computer Administered Self Interviews (ACASI) software was used for sensitive violence questions and Computer-Administered Personal Interviews (CAPI) was used for non-sensitive questions for girls and for the full caregiver survey. Only the older groups of girls (aged 13–14) were asked violence and sexual health modules given ethical concerns.In Ethiopia, ACASI was used for the entire survey due to challenges of hiring female enumerators who can both speak the local non-written languages selected for the study  and read and write in Amharic, Arabic or English. ACASI was selected as a data collection method as it has been shown to increase reporting of sensitive behaviors  in additDescriptive statistics and tests of variables of interest, including chi-square tests and t-tests, will be constructed. Generalized linear mixed models, also known as hierarchal/multilevel mixed models, will be used to assess whether girls in the intervention arm report changes in outcomes of interest compared to girls in the intervention arm, accounting for multiple levels of clustering in the data . Intention-to-treat analyses and per protocol analyses, based on attendance records, will be generated.At baseline, methods of qualitative data collection included focus group discussions with caregivers and social mapping exercises with girls in both countries. In-depth interviews with girls and caregivers were also collected at baseline in DRC only. Similar activities will be conducted at endline. Focus group guides with caregivers sought to identify community gender norms related to girls and key challenges girls may face throughout periods of transitioning to adulthood in their communities, including challenges related to violence. Social mapping activities with girls involved asking girls to draw a map of their community and to note and discuss whether places were safe or unsafe. In-depth interviews with caregivers and girls sought to identify challenges girls face and how members of the family or community can support girls.In DRC, qualitative data collectors were trained to conduct the group activities and record hand-written focus group or mapping notes in either Swahili or Mashi. In-depth interviews with girls and caregivers were audio recorded, transcribed, and then translated into English. In Ethiopia, Ethiopian qualitative data collectors facilitated the group activities in English with assistance from female Berta/Funj or Regarig translators. Only these two languages were selected for the qualitative research in Ethiopia due to challenges locating available female translators who could also speak either English or Amharic to communicate with the qualitative facilitators. All hand-written notes were recorded in English and subsequently transcribed. A combination of inductive content analysis and grounded theory will be utilized for data analysis to generate connections and themes across qualitative data.Three main components comprise the process evaluation for the COMPASS program: (1) attendance records; (2) safe space quality assessments; and (3) service provider quality criteria checklists. All tools have been adapted as needed across countries.First, attendance of girls and caregivers will be systematically tracked in program activities to understand adherence rates and identify any barriers to participation or potential issues with mentors, and follow up with non-attending girls as appropriate.Second, the quality of safe spaces and the quality of programming delivered within the spaces will be assessed through three separate tools. Activities to assess the levels of learning of curriculum topics will be conducted at least twice during the curriculum cycle, which focus on girls’ perceptions of favourite topics, usefulness of different content modules, and any unintended consequences of the programming. A similar method will be used to capture perceptions and knowledge gained from caregivers participating in the program. In addition, mentors will be supported to share their perceptions of girls’ reactions to the programming and the appropriateness of delivery , through supervision meetings. IRC staff will also conduct observations of safe space sessions to provide overall feedback to mentors and to fidelity of session delivery to the curriculum guidelines.The third component for the process evaluation will focus on assessment of service provider quality through training feedback forms, observation reports of case managers and health workers, child client satisfaction surveys, and analysis of GBV Information Management System data to capture the number of survivors receiving services from the IRC or other providers in the study areas.Monitoring data will be used as a mechanism to explain the impact evaluation results, guide per-protocol analyses as findings emerge, and to provide data for other program logical framework indicators that fall outside the scope of the evaluation studies.The COMPASS evaluation will yield important insights into potential strategies to improve the safety of adolescent girls in humanitarian settings by addressing their specific needs and challenges. Importantly, these research studies within COMPASS will assist in unpacking the utility of different approaches, such as caregiver discussion groups to support the empowerment of adolescent girls and reduce girls’ risk of violence. These studies will also demonstrate the potential effectiveness of safe spaces and the programming within safe spaces to improve outcomes specifically related to adolescent girls’ safety, above and beyond children’s protection needs previously examined in safe space research , 33. TheThroughout the inception period and launch of the two randomized controlled trials, a number of challenges emerged and protocols have been adapted accordingly, as detailed below. First, while the original intention of the evaluation was to recruit only unmarried girls, as a key secondary outcome of the study is early marriage, the inclusion criteria had to be loosened. Though all of the girls initially reported themselves to be unmarried in program enrollment forms, which served as the basis for participant recruitment into the studies, they later reported that they were married when asked through ACASI in both DRC and Ethiopia. In Ethiopia, it is thought that this discrepancy was due to the illegal nature of early marriages in Ethiopia  and thusSecond, the diversity of languages was also challenging in the Ethiopian refugee camps. A language assessment was conducted by the program team to determine which languages girls spoke, the level of fluency in each language they were reported to speak, as well as the total number of girls who spoke a language with sufficient fluency that they could understand ACASI in order to ensure sample size requirements could be met. Based on these assessments, as well as a mapping of available and qualified female research assistants or translators that were able to communicate in camp languages, the final four languages were selected for the quantitative survey and two languages for the qualitative activities. While the process behind selecting languages was methodically carried out, some refugee leaders reported the final selection had potential to exacerbate community tensions as only certain languages were selected for inclusion into the research study. Leaders preferred all surveys to have been completed through informal Arabic, which most girls could understand to some degree, but not to the fluency that the survey questions required which would have ultimately limited the reliability and validity of the survey. Importantly, girls speaking any languages will be invited to participate in the program, regardless of whether they are part of the research sample.Other biases have been considered in the study design as well. Primarily, contamination is of concern as intervention groups and wait-listed groups may interact with each other which may minimize our ability to detect changes between groups. Due to sample size requirements for cluster-randomized controlled trials, groups were randomized within communities in DRC and within small geographical areas in refugee camps in Ethiopia. Potential contamination will be assessed during the endline data collection. Social desirability bias may also emerge if respondents in the intervention arm report lower levels of violence, for example, to please the interviewer. However, the use of ACASI may minimize this bias as interviewers will not be aware of the girls’ responses. In addition, another concern is that humanitarian settings are comprised of highly mobile populations, such that girls and caregivers may move out of the study area during the period of program delivery. Attendance will be tracked and intention-to-treat and per-protocol analyses will be undertaken to assess how this attendance may impact the effectiveness of COMPASS. Finally, behavior change to reduce violence against women and girls may require long term investments and our ability to see changes in the limited follow-up period may be minimized. Therefore, examination of secondary outcomes, such as gender attitudes and norms will also be critical to interpretation of the effectiveness of COMPASS as well as the potential salience of different pathways to change. As seen in other community-based trials to reduce intimate partner violence, consistency, trends, and alignment with hypothesized directions of change of both primary and secondary outcomes will be examined to provide insight into potential effectiveness , 36, 37.Despite these challenges, the studies have been strengthened by intensive engagement with in-country program teams as they contextualize the program and research within the political and cultural environments. Rigorous attention to such contextualization by country teams has proven critical for successful engagement with community leaders, adaptation of measures used in the survey, as well as general recruitment of girls and caregivers into the study. Nonetheless, such contextualization, piloting, and attention to translations and language issues extended the research program’s inception and initial planned period of approximately three months to almost one year. This led to a substantial delay in launching the COMPASS programming activities and strained relationships with communities as the period between program registration and launch of activities was lengthened.In conclusion, while undertaking these studies has proven challenging, they provide further evidence that rigorous impact research can be undertaken in humanitarian emergencies on sensitive topics , 38, parAt the time of initial manuscript submission, baseline surveys in Ethiopia and DRC have been completed. The study is ongoing.