Span 20

Genomic DNA was extracted from the cells using the DNA Blood Mini Kit (Qiagen). In the context of CAST, library preparation was performed with a Beckman Biomek FX automated liquid handling system, with 500 ng starting material using SPRIworks HT chemistry (Beckman Coulter). For low-coverage sequencing, samples were prepared with custom 6 base pair barcodes to enable pooling. Library quantification and quality control were performed using a Fragment Analyzer (Advanced Analytics Technologies, Ames, USA). WGS was pursued on an Illumina HiSeq 2500 platform (Illumina, San Diego, USA), using 50 base pair single reads for low-pass sequencing.
Mate-pair DNA library preparation was performed using the Nextera Mate Pair Sample Preparation Kit (Illumina). In brief, 4 μg of high molecular weight genomic DNA was fragmented by the tagmentation reaction in 400 μl, followed by strand displacement. Samples were size-selected to 4–5 kb following the Gel-Plus path of the protocol. A total of 300–550 ng of size-selected DNA was circularized in 300 μl for 16 h at 30°C. After an exonuclease digestion step to get rid of remaining linear DNA, fragmentation to 300–700 bp with a Covaris S2 instrument (LGC Genomics), and binding to streptavidin beads, the libraries were completed via End Repair, A-Tailing, and Illumina Truseq adapter ligation. The final sequencing library was obtained after PCR for 1 min at 98°C, followed by nine cycles of 30 s at 98°C, 30 s at 60°C, 1 min at 72°C, and a final elongation step of 5 min at 72°C. Sequencing was carried out with Illumina HiSeq2000 (2 × 101 bp reads), MiSeq (2 × 75 bp reads), or NextSeq (2 × 150 bp reads) instrument using v3 or v4 chemistry to reach an average spanning coverage of 20–30× for the mate-pair libraries and an average sequencing coverage of 20–30× for deep WGS libraries. After sequencing, the reads were aligned to the hg19 build of the human reference genome using the Illumina-provided alignment software ELAND (version 2) for all RPE-1-derived sequencing data, and using BWA for the cancer genomic data.

Sri Lanka, a multi-ethnic developing country with a mid-year population of 20 million estimated in 2005 and some of the best health and educational indicators in the South Asian region [26] (link), has seen many instances of internal displacement, but these have been characteristically unpredictable and fluid, the main precipitants being the three decades of conflict and the 2004 Tsunami [27] .
The COMRAID cross-sectional survey was carried out in the first half of 2011 in the Kalpitiya division of Puttalam district, North-western province of Sri Lanka. Puttalam district, which has long had a majority Muslim population [28] , [29] , has been an accessible safe haven for a large number of IDPs due to its geographical closeness to conflict areas. The Kalpitiya division (a small peninsular landmass separated from the mainland by a lagoon) was selected for sampling as it had the largest concentration of IDPs within a relatively small geographic area. It is estimated that around 75,000 Muslims (a distinct minority ethnic group in Sri Lanka, whose religion is Islam, and language mainly Tamil) were displaced over a period of few days in 1990 from the northern regions. The majority (72%) were displaced from Mannar district of the Northern Province [28] . According to official sources, Puttalam district received around 15,000 of these internally displaced families (63,000 individuals), mainly arriving via the sea route, who were initially resettled in temporary shelters in the peninsula in 141 locations (mainly welfare centres) [30] , [31] . Figure 1 illustrates the displacement routes and resettlement areas of the northern Muslim IDPs.
Over a time span of 20 years in forced displacement, as the population numbers grew, a proportion of IDPs bought local land, establishing communities based on their pre-displacement communal orientations while others have lived in government welfare centres for the whole duration of displacement. Living circumstances and economic standards are similar across these different types of settlements, and some of the privately established communities in fact function as welfare centres. The 141 welfare camps in existence at the time of the survey were mostly situated in government land and had basic facilities, consisting of permanent, semi-permanent and temporary housing for IDP families numbering from less than hundred families to several hundred. Camps were overseen by a ‘camp officer’ (the village level administrative official) appointed by the government while civil society organizations such as mosque councils were also responsible for handling various communal affairs. The flight of northern Muslims has been well documented in sociological and anthropological studies [32] –[34] ; however, no study to date has investigated the mental health status of these IDPs, considered to be at high risk for developing psychopathology due to the nature of their displacement experience.

The composition of the proposed nanoemulsions (NEs) presented the usual concentrations of sunscreens in photoprotective preparations already described in the literature [8 (link),33 (link)]. Through the use of OMC and EHMC filters to obtain a wide photoprotective spectrum, the concentrations of BO and AV were modulated such that they do not exceed the maximum concentration of a single synthetic filter used, and mainly of the surfactants based on the principle of hydrophilic-lipophilic balance (HLB), which directly influences droplet size and, consequently, the stability and effectiveness of the photoprotective action of nanostructured systems. Two processing methods were evaluated for the preparation of NEs, both of which were high energy, using an ultrasonic processing equipment (US) and an Ultra-Turrax^® high-speed homogenizer (Figure 6). The choice of this type of processing was due to the stability characteristics promoted by the high shear caused by these techniques, resulting in more uniform droplet size distributions when compared with those produced using low-energy methods.
The composition of the formulation developed for preparing the NE is shown in Table 1, with the combination of surfactants being varied to determine the most stable nanoemulsion system.
Synthetic OMC and EHMC chemical filters were used at concentrations of 10% and 3%, respectively, as they are the maximum amounts permitted by current legislation. The 10% BO and AV were added to replace other synthetic filters. The water-soluble surfactants, Tween^® 80 and Span^® 20, were used in the range of 8–20% to evaluate the stability of the proposed nanostructured system. The use of surfactants was based on the HLB calculation ratio. The 1% Aristoflex AVC^® was added to the formulation as a consistency agent.

In the aqueous phase, the HLB value of Tween^® 80 is 15 and that of Span^® 20 is 8.6. After evaluating different concentrations of these surfactants, the best proportion for their application in the preparation of NEs was chosen, which corresponded to a HLB value of 7.25 for the BO in the oil phase. The impact of the variation of the ultrasonic processing parameters in the chosen NEs was verified by varying the processing time (min) and amplitude (NE-A9 to NE-A15). The composition of the NEs and the ultrasound parameters used are presented in Table 3.