Recombinant expression and purification of L2 domain of human epidermal growth factor receptor

Epidermal growth factor receptor (EGFR) is one of the key molecules in cell growth and multiplication and plays an important role in some malignant processes. L2 domain of extra-cellular part of this receptor involved in ligand binding and its inhibition can prevent activation of related signaling pathways. The aim of the present study was cloning and expressing the fragment coding for L2 region of human EGFR for the production of recombinant L2 protein. The total RNA from A431 cells line was extracted and used for amplification of the sequence coding for L2 domain of EGFR by reverse transcriptase-polymerase chain reaction (RT-PCR) technique. The product was cloned into the PGEM-T vector and used for sequencing. In the next step, the insert was removed from the PGEM-T vector and subcloned into the PET22 expression vector. The expression construct was transformed into the Escherichia coli BL21 (DE3) and recombinant protein expression was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Analyzing the expression of produced recombinant protein showed that the recombinant L2 can be highly expressed by this expression system. This recombinant protein can be used for the production of specific mAb, screening for specific ligands and competitive inhibition of the EGFR.Keywords: Human epidermal growth factor receptor, domain L2, recombinant expressionAfrican Journal of Biotechnology Vol. 9(33), pp. 5292-5296, 16 August, 201

Recombinant expression and purification of L2 domain of human epidermal growth factor receptor

Epidermal growth factor receptor (EGFR) is one of the key molecules in cell growth and multiplication and plays an important role in some malignant processes. L2 domain of extra-cellular part of this receptor involved in ligand binding and its inhibition can prevent activation of related signaling pathways. The aim of the present study was cloning and expressing the fragment coding for L2 region of human EGFR for the production of recombinant L2 protein. The total RNA from A431 cells line was extracted and used for amplification of the sequence coding for L2 domain of EGFR by reverse transcriptase-polymerase chain reaction (RT-PCR) technique. The product was cloned into the PGEM-T vector and used for sequencing. In the next step, the insert was removed from the PGEM-T vector and subcloned into the PET22 expression vector. The expression construct was transformed into the Escherichia coli BL21 (DE3) and recombinant protein expression was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Analyzing the expression of produced recombinant protein showed that the recombinant L2 can be highly expressed by this expression system. This recombinant protein can be used for the production of specific mAb, screening for specific ligands and competitive inhibition of the EGFR

Mapping local patterns of childhood overweight and wasting in low- and middle-income countries between 2000 and 2017

A double burden of malnutrition occurs when individuals, household members or communities experience both undernutrition and overweight. Here, we show geospatial estimates of overweight and wasting prevalence among children under 5 years of age in 105 low- and middle-income countries (LMICs) from 2000 to 2017 and aggregate these to policy-relevant administrative units. Wasting decreased overall across LMICs between 2000 and 2017, from 8.4% (62.3 (55.1–70.8) million) to 6.4% (58.3 (47.6–70.7) million), but is predicted to remain above the World Health Organization’s Global Nutrition Target of <5% in over half of LMICs by 2025. Prevalence of overweight increased from 5.2% (30 (22.8–38.5) million) in 2000 to 6.0% (55.5 (44.8–67.9) million) children aged under 5 years in 2017. Areas most affected by double burden of malnutrition were located in Indonesia, Thailand, southeastern China, Botswana, Cameroon and central Nigeria. Our estimates provide a new perspective to researchers, policy makers and public health agencies in their efforts to address this global childhood syndemic

Treatment of atrophic cutaneous leishmaniasis scar using autologous fibroblasts and keratinocytes (a case report and literature review)

In this case report we present a case of wide, deep, atrophic cutaneous leishmaniasis scar that was improved significantly using injection of autologus fibroblasts, dermabrasion and dressing with autologus keratinocytes and fibroblasts. To our best knowledge, it is the first time that separate cultured fibroblasts and keratinocytes are used in conjunction with dermabrasion for treatment of cutaneous leishmaniasis scar. We have started this procedure as an extensive research on patients with leishmaniasis scar that will be reported in the future

Mapping routine measles vaccination in low- and middle-income countries

The safe, highly effective measles vaccine has been recommended globally since 1974, yet in 2017 there were more than 17 million cases of measles and 83,400 deaths in children under 5 years old, and more than 99% of both occurred in low- and middle-income countries (LMICs)1–4. Globally comparable, annual, local estimates of routine first-dose measles-containing vaccine (MCV1) coverage are critical for understanding geographically precise immunity patterns, progress towards the targets of the Global Vaccine Action Plan (GVAP), and high-risk areas amid disruptions to vaccination programmes caused by coronavirus disease 2019 (COVID-19)5–8. Here we generated annual estimates of routine childhood MCV1 coverage at 5 × 5-km2 pixel and second administrative levels from 2000 to 2019 in 101 LMICs, quantified geographical inequality and assessed vaccination status by geographical remoteness. After widespread MCV1 gains from 2000 to 2010, coverage regressed in more than half of the districts between 2010 and 2019, leaving many LMICs far from the GVAP goal of 80% coverage in all districts by 2019. MCV1 coverage was lower in rural than in urban locations, although a larger proportion of unvaccinated children overall lived in urban locations; strategies to provide essential vaccination services should address both geographical contexts. These results provide a tool for decision-makers to strengthen routine MCV1 immunization programmes and provide equitable disease protection for all children

MAPPING LOCAL PATTERNS OF CHILDHOOD OVERWEIGHT AND WASTING IN LOW- AND MIDDLE-INCOME COUNTRIES BETWEEN 2000 AND 2017

A double burden of malnutrition occurs when individuals, household members or communities experience both undernutrition and overweight. Here, we show geospatial estimates of overweight and wasting prevalence among children under 5 years of age in 105 low- and middle-income countries (LMICs) from 2000 to 2017 and aggregate these to policy-relevant administrative units. Wasting decreased overall across LMICs between 2000 and 2017, from 8.4% (62.3 (55.1–70.8) million) to 6.4% (58.3 (47.6–70.7) million), but is predicted to remain above the World Health Organization’s Global Nutrition Target of <5% in over half of LMICs by 2025. Prevalence of overweight increased from 5.2% (30 (22.8–38.5) million) in 2000 to 6.0% (55.5 (44.8–67.9) million) children aged under 5 years in 2017. Areas most affected by double burden of malnutrition were located in Indonesia, Thailand, southeastern China, Botswana, Cameroon and central Nigeria. Our estimates provide a new perspective to researchers, policy makers and public health agencies in their efforts to address this global childhood syndemic