Additional file 1 of The use and potential impact of digital health tools at the community level: results from a multi-country survey of community health workers

Supplementary Material 1

Evaluación del horno de curado de tabaco por convección forzada usco-madr1

Teniendo como base la infraestructura existente de un horno tradicional de curado de tabaco, se rediseño e implementó en él un sistema de intercambio de calor por convección forzada que funciona con cisco de café como combustible. Este horno de curado de tabaco por convección forzada USCO-MADR fue evaluado durante el periodo de cosecha, lográndose un manejo controlado de las variables de temperatura y humedad relativa dentro de él durante las tres etapas del curado de la hoja de tabaco; el equipo utilizado tuvo un excelente desempeño al emplear cisco de café como combustible con los siguientes consumos durante el proceso de curado: en la fase de “amarillamiento”, 8,92 kilogramos por hora; en la de “secado de paño y fijación de color”, 17,75 kilogramos por hora; y en la de “secado de vena”, 19,29 kilogramos por hora; el análisis comparativo de los costos operativos del horno evaluado, con los ajustes propuestos a éste, permiten presentarlo a la cadena de tabaco como una alternativa promisoria.A traditional oven for curing tobacco leaves was redesigned (based on existing infrastructure); a forced-convection heat exchan- ger system was implemented in it which worked with coffee hulls as fuel. This oven (called a forced-convection tobacco leaf curing oven) was evaluated during the harvesting season. It was found that temperature and relative humidity inside the furnace could be controlled with this assembly during the three stages involved in curing tobacco leaves. The equipment used performed excellently when using coffee hulls as fuel, having the following approximate consumption during curing: 8.92 kilograms per hour during the yellowing stage, 17.75 kilograms per hour during the leaf drying and color fixation phase and 19.29 kilograms per hour during the stem drying stage. Comparative analysis of the oven’s operating costs along with the proposed adjustments to be made to it would allow its implementation as a promising alternative in the existing tobacco chain

Generation of lapatinib-resistant (LR) OE19 subclones, and the identification of a novel, acquired <i>Src</i>^E527K mutation.

<p><b>A,</b> Schematic view of the development of lapatinib resistance OE19 cells followed by subcloning of distinct clones of resistant cells for genomic characterization. At right is the listing of somatic alterations identified in the distinct subclones, compared to the genome of the parental OE19 cells. The allelic fraction of each mutation, percent of sequenced reads with the mutant allele, is listed for each candidate mutation. <b>B,</b> IGV (Integrated Genomic Viewer) snapshot of <i>Src</i> mutations in two LR subclones compared to the sequencing seen from this locus in the parental, lapatinib-sensitive cell line. <b>C,</b> Direct sequencing results from genomic DNA from both parental OE19 cells and <i>Src</i>^E527K mutant LR2A and LR2B subclones identifies mutation detected from next-generation sequencing. <b>D,</b> DNA restriction analysis results using <i>Ban</i> II enzyme, from the genomic DNA from the parental OE19, Src E527K mutant LR2A and LR2B subclones, and Het1A (normal esophageal cell line). Ban II enzyme cuts and yields new amplicons of 182 bp in the parental OE19 and Het1A, however, the bands of 203 bp, which contained Src mutants, are still visualized in the two Src mutant subclones, LR2A and LR2B.</p

Impact of ectopic expression of <i>Src</i>^E527K in parental OE19 on lapatinib sensitivity and cell signalling.

<p><b>A,</b> Dose-response curves for lapatinib in the non-transduced parental OE19 (OE19 ^Mock), transduced with GFP (OE19 ^GFP), wild-type <i>Src</i> (OE19 ^{Src wild-type}) or <i>Src</i>^E527K mutation (OE19 ^{Src E527K}). The calculated values of IC₅₀ for lapatinib was >1,000 nM in OE19 ^{Src E527K} cells. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment. The <i>p</i> value calculated by two-way ANOVA was 0.376 for mock <i>vs</i> control (GFP), <0.0001 for control <i>vs</i> wild-type Src, and <0.0001 for control <i>vs Src</i>^E527K. <b>B,</b> Relative cell viability lapatinib in the non-transduced parental OE19 (OE19 ^Mock), transduced with GFP (OE19 ^GFP), wild-type <i>Src</i> (OE19 ^{Src wild-type}) or <i>Src</i>^E527K mutation (OE19 ^{Src E527K}). The <i>p</i> values were calculated by two-tailed <i>t</i>-test. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment. <b>C,</b> Immunoblots showing changes of various signalling proteins after treatment with 1 µM concentration of lapatinib or saracatinib, either alone or combination, in the OE19 cells with or without Src ^E527K transduction.</p

Comparisons of lapatinib sensitivities and the baseline signalling proteins activities between parental and lapatinib-resistant subclones.

<p><b>A,</b> Lapatinib sensitivity curves in the parental OE19 and two lapatinib-resistant (LR) subclones. The calculated values of IC₅₀ of lapatinib were 200 nM in parental cells and >1,000 nM in two LR subclones. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment. The <i>p</i> values were <0.0001 for OE19 <i>vs</i> LR2A and OE19 <i>vs</i> LR2B, and was 0.129 for LR2A <i>vs</i> LR2B. The <i>p</i> values were calculated by two-way ANOVA. <b>B,</b> Immunoblots showing the phosphorylations of distinct signalling molecules in parental OE19 cells and the <i>Src</i>-mutant lapatinib-resistant derivatives.</p

Effects of pharmacologic inhibition of <i>Src</i> using saracatinib in combination with lapatinib in LR subclones.

<p><b>A,</b> Dose-response curves from escalated dose of lapatinib in the presence of 1 µM concentration of saracatinib. The calculated values of IC₅₀ for lapatinib were following; 207.3 nM and 145 nM, respectively, after lapatinib alone and in the presence of saracatinib for parental OE19; >1,000 nM after lapatinib alone for LR2A and LR2B; 91.66 nM and 224.0 nM in the presence of saracatinib in LR2A and LR2B, respectively. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment. The <i>p</i> values calculated by two-way ANOVA were <0.0001 comparing lapatinib alone with lapatinib plus saracatinib both in the LR2A and LR2B. <b>B,</b> Relative cell viability in the parental OE19 and two LR subclones after 1 µM concentration of saracatinib or lapatinib, either alone or combination. The <i>p</i> values were calculated by two-tailed <i>t</i>-test. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment. <b>C,</b> Immunoblots showing changes of in intracellular signalling proteins after treatment with 1 µM concentration of lapatinib or saracatinib, either alone or combination, in LR subclones. Proteins were harvested 4 hours after each treatment. <b>D,</b> Relative cell viability in the OE33, <i>ERBB2</i> and <i>MET</i> co-amplified gastroesophageal cancer cells, after treatment with various drugs either alone or combination. The effect of combination treatment with lapatinib and saracatinib did not differ either from lapatinib alone or saracatinib alone; the combination treatment with lapatinib and crizotinib, which is a potent MET inhibitor, showed synergistic effects. The <i>p</i> values were calculated by two-tailed <i>t</i>-test. Values were presented as relative cellular viability relative to vehicle-treated controls with the mean ± S.E. of quadruplicate from a representative experiment.</p