A COMPARATIVE STUDY TO DETERMINE THE EFFECTS OF VARIABLE CONCENTRATIONS AND DIFFERENT ROUTES OF ADMINISTRATION OF LIDOCAINE IN SUPPRESSING COUGH AND ON HEMODYNAMIC RESPONSE DURING EXTUBATION IN PEDIATRIC AGE GROUP: AN OBSERVATIONAL PROSPECTIVE STUDY

Objectives: The objectives of this study were to compare the effects of different concentrations of lidocaine (2% endotracheal [ET] spray; 10% ET spray; and 2% intravenous [IV]) in suppressing cough and on hemodynamic response during extubation in pediatric age groups; an observational prospective study. Methods: Ninety patients were enrolled for the study and divided into three groups. In Group A, patients were administered (1 mg/kg) of 2% lidocaine ET spray 5 min before extubation; in Group B, patients were administered (1 mg/kg) of 10% lidocaine ET spray 5 min before extubation; and in Group C patients were administered (1 mg/kg) 2% lidocaine IV 3 min before extubation. The three groups were compared for hemodynamic parameters, incidence of cough, breathing pattern, and need for continuous positive airway pressure (CPAP). Results: There was significant attenuation of hemodynamic parameters and less incidence of cough and labored breathing in patient receiving either 10% ET or 2% IV lidocaine. As compared to 2% ET lidocaine, requirement of CPAP support was less in patients who received 10% lidocaine. Patients who were administered 2% IV lidocaine did not receive any CPAP support postextubation. Conclusion: As compared to 2% lidocaine spray postextubation, both 10% lidocaine spray and 2% IV lidocaine postextubation have significantly positive effect on suppression of cough and on hemodynamic parameters

Effects of foliar application of melatonin on gas exchange and certain biochemical characteristics broccoli cv. Palam Samridhi

Considering the rich nutritional status and possibility of broccoli in improving the profitable yield, and wide role of Mel in regulating the plant physiological process, an investigation was carried out at the division of Basic Sciences and Humanities during 2017 to investigate the effect of foliar application of Mel on leaf photosynthetic and biochemical attributes broccoli. Thirty days old and uniform seedlings of broccoli cv. Palam Samridhi were transplanted in the field at a spacing of 45 × 45cm. Different concentrations of Mel, viz. 0, 20, 40, 60 and 80 ppm were sprayed on the plant foliage at 15 days after transplanting (DAT) replicating each treatment four times. Leaf gas exchange and biochemical attributes were tested following the standard procedures. The Results showed the lowest stipulated rate of photosynthesis (10.87 µmole.m-2.sec-1), stomatal conductance (301.44 mole H2O.m-2ses-1) and leaf transpiration (1. 14 mole H2O.m-2ses-1) in untreated plants.  Different doses of Mel significantly increased the values of these attributes and the highest values of photosynthesis (18.63 µmole.m-2.sec-1), stomatal conductance (324.37 mmole.m-2.ses-1) and leaf transpiration (3.23 mmole.m-2.ses-1) with Mel 60 ppm were recorded. The alterations in different biochemical attributes were also evident due to foliar application of Mel and maximum leaf sugar (77.0 and 85.9µg/g), protein (56.9 and 77.3 µg/g), total phenols (260.1 and 339.9 mg/100g), antioxidants (142.8 and 159.9 mg GAE /100g DW) and MSI (94.89 and 97.43 percent) values with Mel 60ppm at 30 and 60DAT, respectively. Therefore, the present study signifies the useful effects of Mel in regulating the physio-biochemical properties of broccoli

Assessment of protein silver nanoparticles toxicity against pathogenic Alternaria solani

Mycogenic synthesis of silver nanoparticles (AgNPs) was carried out in the present investigation using an aqueous extract of endophytic non-pathogenic Alternaria solani F10 (KT721914). The mycosynthesized AgNPs were characterized by means of spectroscopic and microscopic techniques. The surface plasmon resonance found at 430 nm confirmed the formation of stable AgNPs for several weeks at room temperature. Also, the results revealed the formation of spherical and monodispersed AgNPs with an average size of 14.8 +/- 1.2 nm. The FT-IR spectrum suggested that the fungal extracellular proteins and secondary metabolites had the role in Ag reduction and AgNPs capping of which protein Ag nanoconjugates were formed. Furthermore, the mycosynthesized AgNPs exhibited potent antifungal activity against different pathogenic isolates of the same Alternaria solani fungus, the causal pathogen of tomato early blight disease. The antifungal efficiency of the AgNPs at 1, 5 and 10 ppm were evaluated for 8 days after incubation by measuring the inhibition rate of fungal radial growth. The results were further supported by investigating fungal hyphae morphology alteration by scanning and transmission electron microscopy. Treated fungal hyphae showed formation of pits and pores. Also, the mycosynthesized AgNPs were able to pass and distribute throughout the fungal cell area and interact with the cell components.A financial support from European Commission by Erasmus Mundus Scholarship-ACTION 2 WELCOME program is gratefully acknowledged. Work in JAD laboratory was supported by grant BIO2014-54269-R from the Ministerio de Economia y Competividad (Spain).Abdel-Hafez, SII.; Nafady, NA.; Abdel-Rahim, IR.; Shaltout, AM.; Daros Arnau, JA.; Mohamed, MA. (2016). Assessment of protein silver nanoparticles toxicity against pathogenic Alternaria solani. 3 Biotech. 6(199):1-12. https://doi.org/10.1007/s13205-016-0515-6S1126199Abd-Alla MH, Nafady NA, Khalaf DM (2016) Assessment of silver nanoparticles contamination on faba bean-Rhizobium leguminosarum bv. viciae-Glomus aggregatum symbiosis: implications for induction of autophagy process in root nodule. Agric Ecosyst Environ 15(218):163–177Abdel-Hafez SI, Nafady NA, Abdel-Rahim IR, Shaltout AM, Mohamed MA (2016) Biogenesis and optimisation of silver nanoparticles by the endophytic fungus cladosporium sphaerospermum. Int J Nano Chem 2(1):11–19Agrios GN (1997) Plant pathology, 4th edn. Academic Press, LondonAzizi S, Namvar F, Mahdavi M, Ahmad MB, Mohamad R (2013) Biosynthesis of silver nanoparticles using brown marine macroalga, Sargassum muticum aqueous extract. Materials 6(12):5942–5950Birla S, Tiwari V, Gade A, Ingle A, Yadav A, Rai M (2009) Fabrication of silver nanoparticles by Phoma glomerala and its combined effect against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Lett Appl Microbiol 48(2):173–179Chohan S, Perveen R, Mehmood MA, Naz S, Akram N (2015) Morpho-physiological studies, management and screening of tomato germplasm against alternaria solani, the causal agent of tomato early blight. Int J Agric Biol 17(1):111–118Das VL, Thomas R, Varghese RT, Soniya EV, Mathew J, Radhakrishnan EK (2014) Extracellular synthesis of silver nanoparticles by the Bacillus strain CS 11 isolated from industrialized area. 3 Biotech 4(2):121–126Datar VV, Mayee CD (1981) Assessment of losses in tomato yield due to early blight. Indian phytopathol 34:191–195Elyasi M, Khalilzadeh MA, Karimi-Maleh H (2013) High sensitive voltammetric sensor based on Pt/CNTs nanocomposite modified ionic liquid carbon paste electrode for determination of Sudan I in food samples. Food Chem 141(4):4311–4317Ensafi AA, Karimi-Maleh H (2010) Modified multiwall carbon nanotubes paste electrode as a sensor for simultaneous determination of 6-thioguanine and folic acid using ferrocenedicarboxylic acid as a mediator. J Electroanal Chem 640(1):75–83Fayaz M, Tiwary CS, Kalaichelvan PT, Venkatesan R (2010) Blue orange light emission from biogenic synthesized silver nanoparticles using Trichoderma viride. Colloids Surf B Biointerfaces 75(1):175–178Gardes M, Bruns TD (1993) ITS primers with enhanced specificity for basidiomycetes-application to the identification of mycorrhizae and rusts. Mol Ecol 2(2):113–118Gurunathan S, Lee KJ, Kalishwaralal K, Sheikpranbabu S, Vaidyanathan R, Eom SH (2009) Antiangiogenic properties of silver nanoparticles. Biomaterials 30(31):6341–6350Kagithoju S, Godishala V, Nanna RS (2015) Eco-friendly and green synthesis of silver nanoparticles using leaf extract of Strychnos potatorum Linn. F. and their bactericidal activities. 3 Biotech 5(5):709–714Kanmani P, Lim ST (2013) Synthesis and structural characterization of silver nanoparticles using bacterial exopolysaccharide and its antimicrobial activity against food and multidrug resistant pathogens. Process Biochem 48(7):1099–1106Khan MR, Rizvi TF (2014) Nanotechnology: scope and application in plant disease management. Plant Pathol J 13:214–231Khan M, Rizwani GH, Shareef H, Cavar S, Zia-Ul-Haq M (2013) Assessment of total phenolic content and antioxidant potential of methanol extract of Peltophorum pterocarpum (DC.) Backer ex K. Heyne. Pak J Pharm Sci 26(5):967–972Kim KJ, Sung WS, Suh BK, Moon SK, Choi JS, Kim JG, Lee DG (2009a) Antifungal activity and mode of action of silver nano-particles on Candida albicans. Biometals 22(2):235–242Kim SW, Kim KS, Lamsal K, Kim YJ, Kim SB, Jung M, Sim SJ, Kim HS, Chang SJ, Kim JK, Lee YS (2009b) An in vitro study of the antifungal effect of silver nanoparticles on oak wilt pathogen Raffaelea sp. J Microbiol Biotechnol 19(8):760–764Kim SW, Jung JH, Lamsal K, Kim YS, Min JS, Lee YS (2012) Antifungal effects of silver nanoparticles (AgNPs) against various plant pathogenic fungi. Mycobiology 40(1):53–58Kirk AB, Martinelango PK, Tian K, Dutta A, Smith EE, Dasgupta PK (2005) Perchlorate and iodide in dairy and breast milk. Environ Sci Technol 39(7):2011–2017Kumar CG, Sujitha P (2014) Green synthesis of Kocuran-functionalized silver glyconanoparticles for use as antibiofilm coatings on silicone urethral catheters. Nanotechnology 25(32):325101Liu L, Yang J, Xie J, Luo Z, Jiang J, Yang YY, Liu S (2013) The potent antimicrobial properties of cell penetrating peptide-conjugated silver nanoparticles with excellent selectivity for Gram-positive bacteria over erythrocytes. Nanoscale 5(9):3834–3840Loza K, Diendorf J, Sengstock C, Ruiz-Gonzalez L, Gonzalez-Calbet J, Vallet- Regi M, Köller M, Epple M (2014) The dissolution and biological effects of silver nanoparticles in biological media. J Mater Chem B 2:1634–1643Malik P, Shankar R, Malik V, Sharma N, Mukherjee TK (2014) Green chemistry based benign routes for nanoparticle synthesis. J Nanopart 24:1–14McDonnell G, Russell AD (2001) Antiseptics and disinfectants: activity, action, and resistance. Clin Microbiol Rev 14(1):227Metuku RP, Pabba S, Burra S, Gudikandula K, Charya MS (2014) Biosynthesis of silver nanoparticles from Schizophyllum radiatum HE 863742.1: their characterization and antimicrobial activity. 3 Biotech 4(3):227–234Mohamed AM (2015) One-step functionalization of silver Nanoparticles using the orsellinic acid compound isolated from the endophytic fungus Epicoccum Nigrum: characterization and antifungal activity. Int J Nano Chem. 1(3):103–110Moradi R, Sebt SA, Karimi-Maleh H, Sadeghi R, Karimi F, Bahari A, Arabi H (2013) Synthesis and application of FePt/CNTs nanocomposite as a sensor and novel amide ligand as a mediator for simultaneous determination of glutathione, nicotinamide adenine dinucleotide and tryptophan. Phys Chem Chem Phys 15(16):5888–5897Nadworny PL, Wang J, Tredget EE, Burrell RE (2008) Anti-inflammatory activity of nanocrystalline silver in a porcine contact dermatitis model. Nanomedicine 4(3):241–251Namanda S, Olanya OM, Adipala E, Hakiza JJ, El-Bedewy R, Baghsari AS, Ewell P (2004) Fungicide application and host-resistance for potato late blight management: benefits assessment from on-farm studies in SW Uganda. Crop Prot 23(11):1075–1083Narayanan KB, Sakthivel N (2010) Biological synthesis of metal nanoparticles by microbes. Adv Colloid Interface Sci 156(1):1–3Netala VR, Kotakadi VS, Bobbu P, Gaddam SA, Tartte V (2016) Endophytic fungal isolate mediated biosynthesis of silver nanoparticles and their free radical scavenging activity and anti microbial studies. 3 Biotech 6(2):1–9Petrini O, Fisher PJ (1988) A comparative study of fungal endophytes in xylem and whole stems of Pinus sylvestris and Fagus sylvatica. Trans Br Mycol Soc 91(2):233–238Qin Y, Ji X, Jing J, Liu H, Wu H, Yang W (2010) Size control over spherical silver nanoparticles by ascorbic acid reduction. Colloids Surf A Physicochem Eng Asp 372(1):172–176Ramamurthy CH, Padma M, Mareeswaran R, Suyavaran A, Kumar MS, Premkumar K, Thirunavukkarasu C (2013) The extra cellular synthesis of gold and silver nanoparticles and their free radical scavenging and antibacterial properties. Colloids Surf B Biointerfaces 102:808–815Rogers JV, Parkinson CV, Choi YW, Speshock JL, Hussain SM (2008) A preliminary assessment of silver nanoparticle inhibition of monkeypox virus plaque formation. Nanoscale Res Lett 3(4):129–133Sadeghi R, Karimi-Maleh H, Khalilzadeh MA, Beitollahi H, Ranjbarha Z, Zanousi MB (2013) A new strategy for determination of hydroxylamine and phenol in water and waste water samples using modified nanosensor. Environ Sci Pollut Res Int 20(9):6584–6593Saharan V, Sharma G, Yadav M, Choudhary MK, Sharma SS, Pal A, Biswas P (2015) Synthesis and in vitro antifungal efficacy of Cu–chitosan nanoparticles against pathogenic fungi of tomato. Int J Biol Macromolec 75:346–353Satyavani K, Ramanathan T, Gurudeeban S (2011) Plant mediated synthesis of biomedical silver nanoparticles by using leaf extract of Citrullus colocynthis. R J Nanosci Nanotech 1(2):95–101Siddique YH, Fatima A, Jyoti S, Naz F, Khan W, Singh BR, Naqvi AH (2013) Evaluation of the toxic potential of graphene copper nanocomposite (GCNC) in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg 9. PloS one 8(12):e80944Stoimenov PK, Klinger RL, Marchin GL, Klabunde KJ (2002) Metal oxide nanoparticles as bactericidal agents. Langmuir 18(17):6679–6686Tanvir S, Oudet F, Pulvin S, Anderson WA (2012) Coenzyme based synthesis of silver nanocrystals. Enzyme Microb Technol 51(4):231–236Thakkar KN, Mhatre SS, Parikh RY (2010) Biological synthesis of metallic nanoparticles. Nanomedicine 6(2):257–262Vahdati AR, Sadeghi B (2013) A study on the assessment of DNA strand-breaking activity by silver and silica nanoparticles. J Nanostruct Chem 1:1–3Wu D, Fan W, Kishen A, Gutmann JL, Fan B (2014) Evaluation of the antibacterial efficacy of silver nanoparticles against Enterococcus faecalis biofilm. J Endod 40:285–290Zachariadis PC, Hadjikakou SK, Hadjiliadis N, Skoulika S, Michaelides A, Balzarini J, De Clercq E (2004) Synthesis, characterization and in vitro study of the cytostatic and antiviral activity of new polymeric silver (I) complexes with ribbon structures derived from the conjugated heterocyclic thioamide 2-mercapto-3, 4, 5, 6-tetra-hydropyrimidine. Eur J Inorg Chem 7:1420–1426Zhang W, Qiao X, Chen J (2007) Synthesis of silver nanoparticles—effects of concerned parameters in water/oil microemulsion. Mater Sci Eng, B 142(1):1–5Zhao N, Gao J, Enns CA, Knutson MD (2010) ZRT/IRT-like protein 14 (ZIP14) promotes the cellular assimilation of iron from transferrin. J Biol Chem 285(42):32141–3215

Burden of disease scenarios for 204 countries and territories, 2022–2050: a forecasting analysis for the Global Burden of Disease Study 2021

Background: Future trends in disease burden and drivers of health are of great interest to policy makers and the public at large. This information can be used for policy and long-term health investment, planning, and prioritisation. We have expanded and improved upon previous forecasts produced as part of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) and provide a reference forecast (the most likely future), and alternative scenarios assessing disease burden trajectories if selected sets of risk factors were eliminated from current levels by 2050. Methods: Using forecasts of major drivers of health such as the Socio-demographic Index (SDI; a composite measure of lag-distributed income per capita, mean years of education, and total fertility under 25 years of age) and the full set of risk factor exposures captured by GBD, we provide cause-specific forecasts of mortality, years of life lost (YLLs), years lived with disability (YLDs), and disability-adjusted life-years (DALYs) by age and sex from 2022 to 2050 for 204 countries and territories, 21 GBD regions, seven super-regions, and the world. All analyses were done at the cause-specific level so that only risk factors deemed causal by the GBD comparative risk assessment influenced future trajectories of mortality for each disease. Cause-specific mortality was modelled using mixed-effects models with SDI and time as the main covariates, and the combined impact of causal risk factors as an offset in the model. At the all-cause mortality level, we captured unexplained variation by modelling residuals with an autoregressive integrated moving average model with drift attenuation. These all-cause forecasts constrained the cause-specific forecasts at successively deeper levels of the GBD cause hierarchy using cascading mortality models, thus ensuring a robust estimate of cause-specific mortality. For non-fatal measures (eg, low back pain), incidence and prevalence were forecasted from mixed-effects models with SDI as the main covariate, and YLDs were computed from the resulting prevalence forecasts and average disability weights from GBD. Alternative future scenarios were constructed by replacing appropriate reference trajectories for risk factors with hypothetical trajectories of gradual elimination of risk factor exposure from current levels to 2050. The scenarios were constructed from various sets of risk factors: environmental risks (Safer Environment scenario), risks associated with communicable, maternal, neonatal, and nutritional diseases (CMNNs; Improved Childhood Nutrition and Vaccination scenario), risks associated with major non-communicable diseases (NCDs; Improved Behavioural and Metabolic Risks scenario), and the combined effects of these three scenarios. Using the Shared Socioeconomic Pathways climate scenarios SSP2-4.5 as reference and SSP1-1.9 as an optimistic alternative in the Safer Environment scenario, we accounted for climate change impact on health by using the most recent Intergovernmental Panel on Climate Change temperature forecasts and published trajectories of ambient air pollution for the same two scenarios. Life expectancy and healthy life expectancy were computed using standard methods. The forecasting framework includes computing the age-sex-specific future population for each location and separately for each scenario. 95% uncertainty intervals (UIs) for each individual future estimate were derived from the 2·5th and 97·5th percentiles of distributions generated from propagating 500 draws through the multistage computational pipeline. Findings: In the reference scenario forecast, global and super-regional life expectancy increased from 2022 to 2050, but improvement was at a slower pace than in the three decades preceding the COVID-19 pandemic (beginning in 2020). Gains in future life expectancy were forecasted to be greatest in super-regions with comparatively low life expectancies (such as sub-Saharan Africa) compared with super-regions with higher life expectancies (such as the high-income super-region), leading to a trend towards convergence in life expectancy across locations between now and 2050. At the super-region level, forecasted healthy life expectancy patterns were similar to those of life expectancies. Forecasts for the reference scenario found that health will improve in the coming decades, with all-cause age-standardised DALY rates decreasing in every GBD super-region. The total DALY burden measured in counts, however, will increase in every super-region, largely a function of population ageing and growth. We also forecasted that both DALY counts and age-standardised DALY rates will continue to shift from CMNNs to NCDs, with the most pronounced shifts occurring in sub-Saharan Africa (60·1% [95% UI 56·8–63·1] of DALYs were from CMNNs in 2022 compared with 35·8% [31·0–45·0] in 2050) and south Asia (31·7% [29·2–34·1] to 15·5% [13·7–17·5]). This shift is reflected in the leading global causes of DALYs, with the top four causes in 2050 being ischaemic heart disease, stroke, diabetes, and chronic obstructive pulmonary disease, compared with 2022, with ischaemic heart disease, neonatal disorders, stroke, and lower respiratory infections at the top. The global proportion of DALYs due to YLDs likewise increased from 33·8% (27·4–40·3) to 41·1% (33·9–48·1) from 2022 to 2050, demonstrating an important shift in overall disease burden towards morbidity and away from premature death. The largest shift of this kind was forecasted for sub-Saharan Africa, from 20·1% (15·6–25·3) of DALYs due to YLDs in 2022 to 35·6% (26·5–43·0) in 2050. In the assessment of alternative future scenarios, the combined effects of the scenarios (Safer Environment, Improved Childhood Nutrition and Vaccination, and Improved Behavioural and Metabolic Risks scenarios) demonstrated an important decrease in the global burden of DALYs in 2050 of 15·4% (13·5–17·5) compared with the reference scenario, with decreases across super-regions ranging from 10·4% (9·7–11·3) in the high-income super-region to 23·9% (20·7–27·3) in north Africa and the Middle East. The Safer Environment scenario had its largest decrease in sub-Saharan Africa (5·2% [3·5–6·8]), the Improved Behavioural and Metabolic Risks scenario in north Africa and the Middle East (23·2% [20·2–26·5]), and the Improved Nutrition and Vaccination scenario in sub-Saharan Africa (2·0% [–0·6 to 3·6]). Interpretation: Globally, life expectancy and age-standardised disease burden were forecasted to improve between 2022 and 2050, with the majority of the burden continuing to shift from CMNNs to NCDs. That said, continued progress on reducing the CMNN disease burden will be dependent on maintaining investment in and policy emphasis on CMNN disease prevention and treatment. Mostly due to growth and ageing of populations, the number of deaths and DALYs due to all causes combined will generally increase. By constructing alternative future scenarios wherein certain risk exposures are eliminated by 2050, we have shown that opportunities exist to substantially improve health outcomes in the future through concerted efforts to prevent exposure to well established risk factors and to expand access to key health interventions

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Purification and Characterization of Glutaminase Free Asparaginase from Enterobacter cloacae: In-Vitro Evaluation of Cytotoxic Potential against Human Myeloid Leukemia HL-60 Cells.

Asparaginase is an important antileukemic agent extensively used worldwide but the intrinsic glutaminase activity of this enzymatic drug is responsible for serious life threatening side effects. Hence, glutaminase free asparaginase is much needed for upgradation of therapeutic index of asparaginase therapy. In the present study, glutaminase free asparaginase produced from Enterobacter cloacae was purified to apparent homogeneity. The purified enzyme was found to be homodimer of approximately 106 kDa with monomeric size of approximately 52 kDa and pI 4.5. Purified enzyme showed optimum activity between pH 7-8 and temperature 35-40°C, which is close to the internal environment of human body. Monovalent cations such as Na+ and K+ enhanced asparaginase activity whereas divalent and trivalent cations, Ca2+, Mg2+, Zn2+, Mn2+, and Fe3+ inhibited the enzyme activity. Kinetic parameters Km, Vmax and Kcat of purified enzyme were found to be 1.58×10-3 M, 2.22 IU μg-1 and 5.3 × 104 S-1, respectively. Purified enzyme showed prolonged in vitro serum (T1/2 = ~ 39 h) and trypsin (T1/2 = ~ 32 min) half life, which is therapeutically remarkable feature. The cytotoxic activity of enzyme was examined against a panel of human cancer cell lines, HL-60, MOLT-4, MDA-MB-231 and T47D, and highest cytotoxicity observed against HL-60 cells (IC50 ~ 3.1 IU ml-1), which was comparable to commercial asparaginase. Cell and nuclear morphological studies of HL-60 cells showed that on treatment with purified asparaginase symptoms of apoptosis were increased in dose dependent manner. Cell cycle progression analysis indicates that enzyme induces apoptosis by cell cycle arrest in G0/G1 phase. Mitochondrial membrane potential loss showed that enzyme also triggers the mitochondrial pathway of apoptosis. Furthermore, the enzyme was found to be nontoxic for human noncancerous cells FR-2 and nonhemolytic for human erythrocytes