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Item Formulation and stabilization of oil-in-water nanoemulsions using a saponinsrich extract from argan oil press-cake(Food Chemistry, 2010) Taarji, Noamane; A. Rabelo da Silva, Cezar; Nauman Khalid; Gadhi, Chemseddoha; Hafidi, Abdellatif; Kobayashi, Isao; A. Neves, Marcos; Isoda, Hiroko; Nakajima, MitsutoshiIn this study, we formulated and stabilized oil-in-water nanoemulsions using a crude extract from argan press-cake as sole emulsifier. Various extracts from argan press-cake were prepared in order to select the most surface-active one(s) foreseeing emulsions preparation. Fifty percent (v/v) ethanolic extract reduced the interfacial tension to a minimum value at both MCT oil and soybean oil interfaces (12.7 and 10.5 mN m−1 respectively). This extract was also effective at producing fine emulsions with small droplet sizes (d3,2 < 115 nm) and good physical stability using different oils such as soybean oil, MCT oil and fish oil and at conventional homogenization conditions (100 MPa for 4 passes). On the other hand, the emulsions were very sensitive to NaCl addition (≥25 mM) and to acidic pH (<3) indicating that the main stabilization mechanism is electrostatic, likely due to the presence of surface-active compounds with ionizable groups such as saponins.Item Flowering and fruiting responses of strawberry to growth hormone and chilling grown under tunnel conditions.(Pakistan Journal of Agricultural Sciences., 2016) Nauman KhalidStrawberry (Fragaria ananasa) is a berry fruit belonging to the family Rosaceae and requires certain amount of cold temperature to initiate flowering. Present study was conducted with the objective to study flowering and fruiting responses of strawberry to growth hormone and chilling under tunnel conditions. The study comprised of six different treatments as; control, GA3 (gibberellic acid) 200 ppm, GA3 400 ppm, chilling at 4°C, chilling at 4°C + GA3 200 ppm and chilling at 4°C + GA3 400 ppm. Treatments were applied at vegetative stage before flowering. It was inferred from the study that strawberry plants responded much better to application of GA3 at 400 ppm for their growth, flower stimulation, their number, quality features while chilling at 4°C proved better for maximum fruit production and better quality under tunnel condition.Item Controlled assembly of silver nano-fluid in heliotropium crispum extract: a potent anti-biofilm and bactericidal formulation.(applied surface science, Elsevier, 2016) Nauman KhalidThe study describes the optimized method for silver nanoparticle (AgNPs) synthesis using Heliotropium crispum (HC) plant extract. Optimization of physicochemical parameters resulted in stable and rapidly assembled AgNPs. FTIR results suggest presence of plant phytochemicals that helped in the reduction, stabilization and capping of AgNPs. The assembled Ag nano-composites displayed the peak surface plasmon resonance (SPR) around 428 nm. The presence of uniquely assembled Ag-biomolecule composites, cap and stabilize nanoparticles in aqueous plant suspension. Spherical, uniform-shaped AgNPs with low poly-dispersion and average particle size of 42 nm and was determined through dynamic light scattering (DLS) and scanning election microscopy (SEM) which present robust interaction with microbes. The study also evaluates the antimicrobial and anti-biofilm properties of biologically synthesized AgNPs on clinical isolates of MRSA, Pseudomonas aeruginosa and Acinetobacter baumannii. Minimum inhibitory concentration (0.5 mg mL−1) of nanoparticles that presented bactericidal effect was made through inhibition assays on bacterial strains. The concentration which presented potent bactericidal response was then evaluated through growth inhibition in liquid medium for anti-biofilm studies at 2.0 mg mL−1. HC-Ag nanoparticles mediated anti-biofilm effects on Pseudomonas aeruginosa was revealed through SEM. Complete breakdown of biofilm’s extracellular polymeric substances resulted after incubation with AgNPs. Peptidoglycan cell wall destruction was also revealed on planktonic bacterial images after 24 h of incubation.Item Formulation and stability assessment of ergocalciferol loaded oil-in-water nanoemulsions: insights of emulsifiers effect on stabilization mechanism.(Food Research International, Elvsevier, 2016) Nauman KhalidIn the study, we investigated the effect of emulsifiers with different stabilizing mechanisms on the formulation and stability of ergocalciferol loaded oil-in-water (O/W) emulsions. O/W emulsion stabilized by modified lecithin (ML; electrostatic stabilization), sodium caseinate (SC; electrosteric stabilization) or decaglycerol monooleate (MO-7S; steric stabilization) were formulated using high-pressure homogenization. The Sauter mean diameter (d3,2) of emulsions produced by ML, SC and MO-7S were 126 ± 1, 127 ± 4 and 138 ± 3 nm, respectively. The stability of resulting emulsions was evaluated when they exposed to different environmental stresses and during 30 days of storage at 25 and 55 °C. Results showed that the emulsions prepared by MO-7S or ML were stable against a wide range of pH (2–8), while SC-stabilized emulsions showed instability with extensive droplet aggregation at pH 4 or and 5. Only ML-stabilized emulsions showed droplet growth due to coalescence when treated at high NaCl concentration (300–500 mM). In the absence of glucose, SC-stabilized O/W emulsions showed better freeze-thaw stability, in comparison to those formed with ML or MO-7S emulsifiers. The emulsion produced by ML was found to be stable to droplet aggregation at heating temperatures (80–120 °C) for 1 h. All the O/W emulsions stored at 25 °C showed good physical and chemical stability. However, the chemical stability of ergocalciferol in emulsion system decreased in order of ML > MO-7S ≫ SC during storage at 55 °C for a period of 30 days. These findings provide valuable information for the development of nanoemulsion-based delivery system applied in food and beverage products.Item Formulation and characterization of esterified xylo-oligosaccharides-stabilized oil-in-water emulsions using microchannel emulsification.(Colloids and Surfaces B-Biointerfaces, Elsevier, 2016) Nauman KhalidAseries of amphiphilically esterified xylo-oligosaccharides (xylo esters) with differentfatty acids residues – decanoic acid (C-10), lauric acid (C-12) and palmitic acid (C-16) – were enzymatically modified at 60 ◦C for 4 h. These xylo esters were used as emulsifiers to formulate oil-in-water (O/W) emulsions by microchannel emulsification (MCE). Grooved and straight-through MCE was used to investigate the droplet generation and/or emulsion stability. Xylo ester-stabilized oil droplets were generated smoothly from microchannels arranged linearly or two dimensionally, while xylo ester-stabilized emulsions were less monodispersed owing to low surface activity of the xylo esters. The combined use of xylo esters (2.5% (w/w)) and Tween series (0.1% (w/w)) in the continuous phase can improve the monodispersity of the resultant oil. Successful droplet generation was achieved with the straight-through MCE using 2.5% (w/w) xylo laurate and 0.1% (w/w) Tween 20. The optimized combination of xylo laurate and Tween 20 inhibited coalescence and oiling off more efficiently than the droplets solely stabilized by Tween 20 during 30 days of storage.Item Biodegradation of phenol by stenotrophomonas sp. And staphylococcus sp. Isolated from contaminated sites.(Applied Ecology and Enviromental Research, 2016) Nauman KhalidPhenol as environmental pollutant is detrimental to living organisms and needed to be eliminated for environmental safety. Among the various practiced approaches for its removal, bacterial utilization gets attraction due to its eco-friendly and cost effective nature. For this purpose, bacterial strains were isolated from bioremediation site and industrial waste through enrichment in phenol (250 mg L -1 ) for 3 days at 28oC. After enrichment, morphologically distinct colonies were purified on phenol (200 mg L-1 ) agar plates and the strains were identified through 16S rRNA gene sequence. Total of eight strains were identified, among them two strains, NCCP-310 and NCCP-405 had the best potential of phenol degradation which were identified as the members of the genera Stenotrophomonas and Staphylococcus. NCCP-310 and NCCP- 405 showed 98.85 and 98.9% sequence identity with Stenotrophomonas maltophilia and Staphylococcus equorum subsp. equorum, respectively. Both strains have ability to tolerate 1000 mg L-1 phenol. The isolated strains degraded 750 mg L-1 of phenol at pH 7 and 28+2oC. NCCP-310 and NCCP-405 showed degradation of such amount in 65 and 85 h with the average rate of 15.65 and 11.64 mg L-1 h -1 . Our work suggests that these strains are efficient in phenol removal and could be used for bioremediation.Item Microchannel emulsification study on formulation and stability characterization of monodisperse oil-in-water emulsions encapsulating quercetin.(Food Chemistry, Elsevier, 2016) Nauman KhalidThe study used microchannel emulsification (MCE) to encapsulate quercetin in food grade oil-in-water (O/W) emulsions. A silicon microchannel plate (Model WMS 1-2) comprised of 10,300 discrete 10 × 104 μm microslots was connected to a circular microhole with an inner diameter of 10 μm. 1% (w/w) Tween 20 was used as optimized emulsifier in Milli-Q water, while 0.4 mg ml−1 quercetin in different oils served as a dispersed phase. The MCE was carried by injecting the dispersed phase at 2 ml h−1. Successful emulsification was conducted below the critical dispersed phase flux, with a Sauter mean diameter of 29 μm and relative span factor below 0.25. The O/W emulsions remained stable in terms of droplet coalescence at 4 and 25 °C for 30 days. The encapsulation efficiency of quercetin in the O/W emulsions was 80% at 4 °C and 70% at 25 °C during the evaluated storage period.Item Microchannel emulsification: a promising technique towards encapsulation of functional compounds(Critical Reviews in Food Science and Nutrition, 2018) Nauman Khalid; Kobayashi, Isao; A. Neves, Marcos; Uemura, Kunihiko; Nakajima, MitsutoshiThis review provides an overview of microchannel emulsification (MCE) for production of functional monodispersed emulsion droplets. The main emphasis has been put on functional bioactives encapsulation using grooved-type and straight-through microchannel array plates. MCE successfully encapsulates the bioactives like β-carotene, oleuropein, γ-oryzanol, β-sitosterol, L-ascorbic acid and ascorbic acid derivatives, vitamin D and quercetin. These bioactives were encapsulated in a variety of delivery systems like simple and multiple emulsions, polymeric particles, microgels, solid lipid particles and functional vesicles. The droplet generation process in MCE is based upon spontaneous transformation of interfaces rather than high energy shear stress systems. The scale-up of MCE can increase the productivity of monodispersed droplets >100 L h−1 and makes it a promising tool at industrial level.Item Emulsion stability of clove oil in chitosan and sodium alginate matrix(International Journal of Food Properties, 2018) Purwanti, Nanik; Syahrian Zehn, Azmi; Dian Pusfitasari, Eka; Nauman Khalid; Yundra Febrianto, Erfin; Suro Mardjan, Sutrisno; Andreas & Isao KobayashiClove oil was emulsified in 1% w/w chitosan (CC emulsions) and 2.5% w/w sodium alginate matrix (CA emulsions) containing Tween 80 as the surfactant. Different homogenization speeds (5,000, 10,000, 15,000 and 20,000 rpm) were used to produce the emulsions, and the stability of the emulsions during storage (29 days) was determined. The stability of the emulsions containing clove oil prior to the solidification process was assessed when chitosan and sodium alginate were used as encapsulating materials. Different homogeniza- tion speeds resulted in polydisperse emulsions with a size of 2–3μm and 90% of stability after 29 days of storage. Different homogenization speeds did not significantly affect the concentrations of the active compounds contained in the emulsions. However, these concentrations changed significantly after 29 days of storage when sodium alginate was used to make the emulsions and the homogenization speeds were≥10,000 rpm. High temperature caused by the high viscosity of the solution and high energy dissipation during homogenization suggested that the emulsions composed of sodium alginate were unstable. Chitosan enabled a longer processing time during the clove oil encapsulation process compared to sodium alginate, when emulsification by homogenization was used. The stability of the emulsion of the clove oil-in- chitosan matrix prior to the solidification step was superior.Item A question mark on emerging zinc-related nutritional deficiencies in Pakistani population(Asia-Pacific Journal of Public Health, 2018) Waqas Asghar; Wahab Nazir; Nauman KhalidItem Global, regional, and national age-sex-specific mortality and life expectancy, 1950-2017: a systematic analysis for the global burden of disease study 2017(Lancet, 2018) Nauman KhalidBackground Assessments of age-specific mortality and life expectancy have been done by the UN Population Division, Department of Economics and Social Affairs (UNPOP), the United States Census Bureau, WHO, and as part of previous iterations of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD). Previous iterations of the GBD used population estimates from UNPOP, which were not derived in a way that was internally consistent with the estimates of the numbers of deaths in the GBD. The present iteration of the GBD, GBD 2017, improves on previous assessments and provides timely estimates of the mortality experience of populations globally. Methods The GBD uses all available data to produce estimates of mortality rates between 1950 and 2017 for 23 age groups, both sexes, and 918 locations, including 195 countries and territories and subnational locations for 16 countries. Data used include vital registration systetns, sample registration systetns, household surveys (complete birth histories, summary birth histories, sibling histories), censuses (summary birth histories, household deaths), and Demographic Surveillance Sites. In total, this analysis used 8259 data sources. Estimates of the probability of death between birth and the age of 5 years and between ages 15 and 60 years are generated and then input into a model life table system to produce complete life tables for all locations and years. Fatal discontinuities and mortality due to HIV/AIDS are analysed separately and then incorporated into the estimation. We analyse the relationship between age-specific mortality and development status using the Socio-demographic Index, a composite measure based on fertility under the age of 25 years, education, and income. There are four main methodological improvements in GBD 2017 compared with GBD 2016: 622 additional data sources have been incorporated; new estimates of population, generated by the GBD study, are used; statistical methods used in different components of the analysis have been further standardised and improved; and the analysis has been extended backwards in time by two decades to start in 1950. Findings Globally, 18.7% (95% uncertainty interval 18.4-19.0) of deaths were registered in 1950 and that proportion has been steadily increasing since, with 58.8% (58.2-59.3) of all deaths being registered in 2015. At the global level, between 1950 and 2017, life expectancy increased from 48.1 years (46.5-49.6) to 70.5 years (70.1-70.8) for men and from 52.9 years (51.7-54.0) to 75.6 years (75.3-75.9) for women. Despite this overall progress, there remains substantial variation in life expectancy at birth in 2017, which ranges from 49.1 years (46.5-51.7) for men in the Central African Republic to 87.6 years (86.9-88.1) among women in Singapore. The greatest progress across age groups was for children younger than 5 years; under-5 mortality dropped from 216.0 deaths (196.3-238.1) per 1000 livebirths in 1950 to 38.9 deaths (35.6-42.83) per 1000 livebirths in 2017, with huge reductions across countries. Nevertheless, there were still 5.4 million (5.2-5.6) deaths among children younger than 5 years in the world in 2017. Progress has been less pronounced and more variable for adults, especially for adult tnales, who had stagnant or increasing mortality rates in several countries. The gap between male and female life expectancy between 1950 and 2017, while relatively stable at the global level, shows distinctive patterns across super-regions and has consistently been the largest in central Europe, eastern Europe, and central Asia, and smallest in south Asia. Performance was also variable across countries and time in observed mortality rates compared with those expected on the basis of development. Interpretation This analysis of age-sex-specific mortality shows that there are remarkably complex patterns in population mortality across countries. The findings of this study highlight global successes, such as the large decline in under-5 mortality, which reflects significant local, national, and global commitment and investment over several decades. However, they also bring attention to mortality patterns that are a cause for concern, particularly among adult men and, to a lesser extent, wotnen, whose mortality rates have stagnated in many countries over the time period of this study, and in some cases are increasing. Copyright C) 2018 The Author(s). Published by Elsevier Ltd.Item Pharmacological and phytochemical analysis of bergenia ciliata leaf and rhizome extracts(Pakistan Journal of Pharmaceutical Sciences, 2018) Saiqa Yousaf; Ghazala Kaukab; Hina Gul; Nauman Khalid; Rehana Kausar; Haroon Ahmed; Huma Ajab; Muhammad GulfrazAntibacterial, antifungal, antioxidant, cytotoxic, and anti-haemolytic activity of various rhizome extracts of Bergenia ciliata were evaluated in this study. The results showed inhibition of the growth of all selected bacterial and fungal strains in comparison with standard antibiotics. The antioxidant activities of Bergenia ciliata extracts were evaluated against DPPH, H2O2, ABTS, total antioxidant capacity and reducing power assays. The order of antioxidant activity of various extracts were methanol> ethanol>n-hexane> aqueous>chloroform. The cytotoxicity ( brine shrimp assay) and anti-haemolytic activities of plant extracts were also promising and varies in dose depended manner. The phytochemical analysis of rhizome extracts of Bergenia ciliata revealed presence of various secondary metabolites which might be responsible for the antimicrobial, antioxidant, cytotoxic and anti-haemolytic activities.Item Long-term stability of droplet production by microchannel (step) emulsification in microfluidic silicon chips with large number of terraced micro channels(Chemical Engineering Journal, 2018) T. Vladisavljevi, Goran; E. Ekanem, Ekanem; Zhang, Zilin; Nauman Khalid; Kobayashi, Isao; Nakajima, MitsutoshiA long-term production stability of oil-in-water emulsions by microchannel (step) emulsification was investigated using two cross flow silicon chips consisting of 540 or 1850 microchannels fabricated on 10 parallel terraces. Each terrace was 9.54 mm long and consisted of 54 channels with a depth of 5 μm and a width of 18 μm (chip 1) or 185 channels with a depth of 4 μm and a width of 8 μm (chip 2). The dispersed phase was a mixture of 2 wt% polycaprolactone (PCL) and 1 wt% poly(d,l-lactic) acid (PLA) dissolved in dichloromethane and the continuous phase was 2 wt% poly(vinyl alcohol). After solvent evaporation, the droplets were transformed into highly uniform composite polymer particles with an average diameter of 8.8 or 4.9 μm. The percentages of active channels, droplet sizes and droplet generation frequencies at individual terraces were investigated in both chips to reveal any flow maldistribution. After 6 h of production in chip 1, overall 95% of the channels produced droplets and the percentage of active channels on each terrace was at least 91%. The percentage of active channels decreased by decreasing the size of the channels. The mean droplet size varied negligibly across individual terraces over 7 h. Slightly higher droplet generation frequencies and smaller percentages of active channels were observed at central terraces. The droplet generation frequencies at the channels located close to each other were similar, indicating that droplet formation dynamics was coupled. The droplet size was not affected by the dispersed phase flow rate.Item Formulation and characterization of astaxanthin-enriched nanoemulsions stabilized using ginseng saponins as natural emulsifiers(Food Chemistry, 2018) Shu, Gaofeng; Nauman Khalid; Chen, Zhang; A. Neves, Marcos; J. Barrow, Colin; Nakajima, MitsutoshiIn this study ginseng saponins (GS) were used as natural emulsifiers to formulate and stabilize O/W nanoemulsions loaded with astaxanthin (AST). GS were found to be highly effective at reducing the interfacial tension at the soybean oil–water interfaces, and were capable of producing nano-scaled droplets (d4,3 ≈ 125 nm) using a high-pressure homogenizer. The droplet size of the nanoemulsions decreased with increasing emulsifier concentration and homogenization pressure. The nanoemulsions were stable without droplet coalescence against thermal treatment (30–90 °C, 30 min), and over a narrow range of pH values (7–9). GS-coated droplets were unstable in acidic conditions (pH 3–6) and in the presence of salt (>25 mM NaCl). The formulated nanoemulsions showed slight change in d4,3 during 15 days of storage at 5, 25 and 40 °C. However, the chemical stability strongly depended on the storage temperature, with the lowest level of AST retained in nanoemulsions stored at higher temperature.Item Measuring progress from 1990 to 2017 and projecting attainment to 2030 of the health-related sustainable development goals for 195 countries and territories: a systematic analysis for the global burden of disease study 2017(Lancet, 2018) Nauman KhalidBackground: Efforts to establish the 2015 baseline and monitor early implementation of the UN Sustainable Development Goals (SDGs) highlight both great potential for and threats to improving health by 2030. To fully deliver on the SDG aim of “leaving no one behind”, it is increasingly important to examine the health-related SDGs beyond national-level estimates. As part of the Global Burden of Diseases, Injuries, and Risk Factors Study 2017 (GBD 2017), we measured progress on 41 of 52 health-related SDG indicators and estimated the health-related SDG index for 195 countries and territories for the period 1990–2017, projected indicators to 2030, and analysed global attainment.Item Global, regional, and national disability-adjusted life-years (dalys) for 359 diseases and injuries and healthy life expectancy (hale) for 195 countries and territories, 1990-2017: a systematic analysis for the global burden of disease study 2017(Lancet, 2018) Nauman KhalidBackground: How long one lives, how many years of life are spent in good and poor health, and how the population’s state of health and leading causes of disability change over time all have implications for policy, planning, and provision of services. We comparatively assessed the patterns and trends of healthy life expectancy (HALE), which quantifies the number of years of life expected to be lived in good health, and the complementary measure of disability-adjusted lifeyears (DALYs), a composite measure of disease burden capturing both premature mortality and prevalence and severity of ill health, for 359 diseases and injuries for 195 countries and territories over the past 28 years.Item In silico functional and tumor suppressor role of hypothetical protein pcnxl2 with regulation of the notch signaling pathway(Rsc Advances, 2018) Muhammad Naveed; Komal Imran; Ayesha Mushtaq; Abdul Sammad Mumtaz; Hussain A. Janjua; Nauman KhalidSince the last decade, various genome sequencing projects have led to the accumulation of an enormous set of genomic data; however, numerous protein-coding genes still need to be functionally characterized. These gene products are called “hypothetical proteins”. The hypothetical protein pecanex-like protein 2 Homo sapiens (PCNXL2) is found to be mutated in colorectal carcinoma with microsatellite instability; therefore, annotation of the function of PCNXL2 in tumorigenesis is very important. In the present study, bioinformatics analysis of PCNXL2 was performed at the molecular level to assess its role in the progression of cancer for designing new anti-cancer drugs. The retrieved sequence of PCNXL2 was functionally and structurally characterized through the web tools Pfam, Batch CD (conserved domain) search, ExPASy, COACH and I-TASSER directed for pathway analysis and design to explore the intercellular interactions of PCNXL2 involved in cancer development. The present study has shown that PCNXL2 encodes multi-pass transmembrane proteins whose tumor suppressor function may involve regulating Notch signaling by transporting protons across the membrane to provide suitable membrane potential for γ secretase function, which may liberate the Notch intracellular domain NICD from the receptor to inside the cell. Furthermore, domain A of PCNXL2 may exhibit nuclear transport activity of NICD from the cytoplasm to the nucleus through interaction with a nuclear localization signal that may act as an activator for Notch signaling in the nucleus. Conclusively, the tumor suppressor role of PCNXL2 by regulation of the Notch signaling pathway and its functional and structural characteristics are important findings. However, further studies are required to validate the putative role of PCNXL2 as a cancer biomarker in cancer development.Item Population and fertility by age and sex for 195 countries and territories, 1950-2017: a systematic analysis for the global burden of disease study 2017(Lancet, 2018) Nauman KhalidPopulation estimates underpin demographic and epidemiological research and are used to track progress on numerous international indicators of health and development. To date, internationally available estimates of population and fertility, although useful, have not been produced with transparent and replicable methods and do not use standardised estimates of mortality. We present single-calendar year and single-year of age estimates of fertility and population by sex with standardised and replicable methods.Item In vitro bio accessibility of ergocalciferol in nanoemulsion-based delivery system: the influence of food-grade emulsifiers with different stabilising mechanisms(International Journal of Food Science and Technology, 2018) Shu, Gaofeng; Nauman Khalid; Boon Tan, Tai; Zhao, Yiguo; A. Neves, Marcos; Kobayashi, Isao; Nakajima, MitsutoshiThe effect of emulsifier type on thein vitrobioaccessibility of ergocalciferol-loaded nanoemulsions wasexamined (mouth, stomach and small intestinal phases). Oil-in-water nanoemulsions were prepared usingemulsifiers with different stabilising mechanisms: decaglycerol monooleate (MO7S; steric), modifiedlecithin (ML; electrostatic), sodium caseinate (SC; electrosteric) and ML-MO7S (combined electrostaticand steric). The droplet size, size distribution,f-potential and microstructure of nanoemulsions duringdigestion depended on the emulsifier type. The fate of lipid in the small intestinal phase also relied on theemulsifier type, with the free fatty acids release rate decreasing in the following order: MO7S>ML-MO7S>ML>SC. The ergocalciferol bioaccessibilities in nanoemulsions prepared using MO7S (62%),ML (64%) and ML-MO7S (65%) were similar and significantly higher than that stabilised by SC (12%).No significant loss of ergocalciferol was observed in all nanoemulsions after full digestion; they werechemically stable against digestion conditions, regardless of the emulsifier type.Item Global, regional, and national comparative risk assessment of 84 behavioral, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990-2017: a systematic analysis for the global burden of disease study 2017(Lancet, 2018) Nauman KhalidBackground The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017 comparative risk assessment (CRA) is a comprehensive approach to risk factor quantification that offers a useful tool for synthesizing evidence on risks and risk–outcome associations. With each annual GBD study, we update the GBD CRA to incorporate improved methods, new risks and risk–outcome pairs, and new data on risk exposure levels and risk–outcome associations.