Plant glucosinolate biosynthesis and breakdown pathways shape the rhizosphere bacterial/archaeal community.

Rhizosphere microbial community assembly results from microbe-microbe-plant interactions mediated by small molecules of plant and microbial origin. Studies with Arabidopsis thaliana have indicated a critical role of glucosinolates in shaping the root and/or rhizosphere microbial community, likely through breakdown products produced by plant or microbial myrosinases inside or outside of the root. Plant nitrile-specifier proteins (NSPs) promote the formation of nitriles at the expense of isothiocyanates upon glucosinolate hydrolysis with unknown consequences for microbial colonisation of roots and rhizosphere. Here, we generated the A. thaliana triple mutant nsp134 devoid of nitrile formation in root homogenates. Using this line and mutants lacking aliphatic or indole glucosinolate biosynthesis pathways or both, we found bacterial/archaeal alpha-diversity of the rhizosphere to be affected only by the ability to produce aliphatic glucosinolates. In contrast, bacterial/archaeal community composition depended on functional root NSPs as well as on pathways of aliphatic and indole glucosinolate biosynthesis. Effects of NSP deficiency were strikingly distinct from those of impaired glucosinolate biosynthesis. Our results demonstrate that rhizosphere microbial community assembly depends on functional pathways of both glucosinolate biosynthesis and breakdown in support of the hypothesis that glucosinolate hydrolysis by myrosinases and NSPs happens before secretion of products to the rhizosphere.

Do High School Students in India Gamble? A Study of Problem Gambling and Its Correlates.

Studies from the West suggest that significant numbers of high school students gamble, despite it being illegal in this age group. To date, there have been no studies on the prevalence of gambling among senior high school and higher secondary school students in India. This study reports point prevalence of gambling and its psychosocial correlates among high school students in the State of Kerala, India. 5043 high school students in the age group 15-19 years, from 73 schools, were selected by cluster random sampling from the district of Ernakulam, Kerala, South India. They completed questionnaires that assessed gambling, substance use, psychological distress, suicidality, and symptoms of Attention Deficit Hyperactivity Disorder (ADHD). Of a total of 4989 completed questionnaires, 1400 (27.9 %) high school students reported to have ever gambled and 353 (7.1 %) were problem gamblers. Of those who had ever gambled, 25.2 % were problem gamblers. Sports betting (betting on cricket and football) was the most popular form of gambling followed by the lottery. Problem gamblers when compared with non-problem gamblers and non-gamblers were significantly more likely to be male, have academic failures, have higher rates of lifetime alcohol and tobacco use, psychological distress, suicidality, history of sexual abuse and higher ADHD symptom scores. Gambling among adolescents in India deserves greater attention, as one in four students who ever gambled was a problem gambler and because of its association with a range of psychosocial variables.

Prevalence & correlates of tobacco use among adolescents in Kerala, India.

BACKGROUND & OBJECTIVES: Most tobacco users who initiate its use during adolescence are likely to continue the use into adulthood and contribute to the 90 per cent of premature deaths among tobacco users. In this context the prevalence, patterns and correlates of tobacco use were studied among adolescent school students in Kerala, India. METHODS: Total 7560 students from classes 8, 10 and 12, within the age group of 12-19 yr, across 73 schools in Ernakulam district, Kerala, India, selected by cluster random sampling, completed a self-administered questionnaire incorporating standardized instruments. RESULTS: Of the 7350 valid questionnaires, the overall lifetime prevalence of tobacco use was 6.9 per cent (12.5% males and 1.2% females). The prevalence of tobacco use increased from 3.1 per cent at 12-13 yr to 15.1 per cent at 18-19 yr. The mean age of onset of tobacco use was 14.0±2.2 yr. The prevalence was higher among students from urban backgrounds, lower socio-economic status and those with part-time jobs. Tobacco users had significantly higher rates of use of alcohol (67.8 vs. 11%) and illicit drugs (33 vs. 6.1%). They had poorer academic performance (24.7 vs. 9.1%), more severe psychological distress (10.8 vs. 4.5%), suicidal attempts (10.2 vs. 3.5%), higher scores of ratings of attention deficit hyperactivity disorder (8.3 vs. 2.5%) and history of sexual abuse (12.5 vs. 3.8%). INTERPRETATION & CONCLUSIONS: The prevalence of tobacco use in adolescents reported in this study was relatively lower than those reported from other Indian States. However, it correlates with multiple negative outcomes suggesting a need to promote specific interventions to prevent adolescent tobacco use.

An epidemological study of obsessive compulsive disorder in adolescents from India.

OBJECTIVES: There is scarce data on the prevalence of OCD among adolescents in India. This study reports point prevalence of OCD among school students (age 12-18years) in the Kerala state of India and examines its association with ADHD, psychological distress, tobacco/alcohol abuse, suicide risk and history of sexual abuse. METHOD: 7560 students of 73 schools were self-administered the OCD subsection of Clinical Interview Schedule-Revised, the Composite International Diagnostic Interview (CIDI) for obsessive compulsive symptoms and other relevant instruments to identify OCD and related clinical measures. A diagnosis of ICD-10 OCD was derived through the CIS-R algorithm which required duration of at least 2weeks and at least a thought/behavior to be resisted along with a cut-off score for severity and impairment. RESULTS: In the sample, 50.3% were males with a mean age of 15.2years (range of 12-18years). The response rate was 97.3% (7380 valid responses). 0.8% (n=61) fulfilled criteria for OCD with a male predominance (1.1 vs. 0.5%, p=0.005). Prevalence was higher among Muslims and increased with age. Taboo thoughts (62.3%) and mental rituals (45.9%) were the commonest symptoms. Those with OCD had significantly higher suicidal thoughts (59 vs. 16.3%, p<0.01) suicide attempts (24.6 vs. 3.8%, p<0.01), ADHD (28 vs. 4%, p<0.001), sexual abuse (24.6 vs. 4.2%, p<0.01), and tobacco use (23 vs. 6.8%, p=0.01). They also reported greater psychological distress and poorer academic performance. CONCLUSIONS: OCD is common among adolescents in India. Its associations with ADHD, sexual abuse, psychological distress, poorer academic performance and suicidal behavior are additional reasons for it to be recognized and treated early.

Age-associated bidirectional modulation of gene expression in single identified R15 neuron of Aplysia.

BACKGROUND: Despite the advances in our understanding of aging-associated behavioral decline, relatively little is known about how aging affects neural circuits that regulate specific behaviors, particularly the expression of genes in specific neural circuits during aging. We have addressed this by exploring a peptidergic neuron R15, an identified neuron of the marine snail Aplysia californica. R15 is implicated in reproduction and osmoregulation and responds to neurotransmitters such as acetylcholine, serotonin and glutamate and is characterized by its action potential bursts. RESULTS: We examined changes in gene expression in R15 neurons during aging by microarray analyses of RNAs from two different age groups, mature and old animals. Specifically we find that 1083 ESTs are differentially regulated in mature and old R15 neurons. Bioinformatics analyses of these genes have identified specific biological pathways that are up or downregulated in mature and old neurons. Comparison with human signaling networks using pathway analyses have identified three major networks [(1) cell signaling, cell morphology, and skeletal muscular system development (2) cell death and survival, cellular function maintenance and embryonic development and (3) neurological diseases, developmental and hereditary disorders] altered in old R15 neurons. Furthermore, qPCR analysis of single R15 neurons to quantify expression levels of candidate regulators involved in transcription (CREB1) and translation (S6K) showed that aging is associated with a decrease in expression of these regulators, and similar analysis in three other neurons (L7, L11 and R2) showed that gene expression change during aging could be bidirectional. CONCLUSIONS: We find that aging is associated with bidirectional changes in gene expression. Detailed bioinformatics analyses and human homolog searches have identified specific biological processes and human-relevant signaling pathways in R15 that are affected during aging. Evaluation of gene expression changes in different neurons suggests specific transcriptomic signature of single neurons during aging.

Dynamic ensemble view of the conformational landscape of HIV-1 TAR RNA and allosteric recognition.

RNA conformational dynamics and the resulting structural heterogeneity play an important role in RNA functions, e.g., recognition. Recognition of HIV-1 TAR RNA has been proposed to occur via a conformational capture mechanism. Here, using ultrafast time-resolved fluorescence spectroscopy, we have probed the complexity of the conformational landscape of HIV-1 TAR RNA and monitored the position-dependent changes in the landscape upon binding of a Tat protein-derived peptide and neomycin B. In the ligand-free state, the TAR RNA samples multiple families of conformations with various degrees of base stacking around the three-nucleotide bulge region. Some subpopulations partially resemble those ligand-bound states, but the coaxially stacked state is below the detection limit. When Tat or neomycin B binds, the bulge region as an ensemble undergoes a conformational transition in a position-dependent manner. Tat and neomycin B induce mutually exclusive changes in the TAR RNA underlying the mechanism of allosteric inhibition at an ensemble level with residue-specific details. Time-resolved anisotropy decay measurements revealed picosecond motions of bases in both ligand-free and ligand-bound states. Mutation of a base pair at the bulge--stem junction has differential effects on the conformational distributions of the bulge bases. A dynamic model of the ensemble view of the conformational landscape for HIV-1 TAR RNA is proposed, and the implication of the general mechanism of RNA recognition and its impact on RNA-based therapeutics are discussed.

Prevalence and Correlates of Self-Reported ADHD Symptoms in Children Attending School in India.

Objective: To study the prevalence and correlates of self-reported ADHD symptoms among school-going adolescents from Kerala, India. Method: Seven thousand five hundred sixty students from Classes 8, 10, and 12, aged 12 to 19 years, across 73 schools selected by cluster random sampling, were invited to participate, but only 7,150 successfully completed the questionnaire incorporating standardized instruments. Results: Three hundred five (4.3%) self-reported symptoms for ADHD combined type, 131 (1.8%) for ADHD hyperactive-impulsive type, and 102 (1.4%) for ADHD inattentive type with a male predominance. Binary logistic regression analysis showed that those with symptoms of ADHD (combined type) compared with the non-ADHD group had poorer academic performance, significantly higher substance use, psychological distress, suicidality, and sexual abuse. Conclusion: The high prevalence of self-reported ADHD symptoms and its association with negative correlates previously reported in literature in those with a diagnosis of ADHD suggests that clinically significant self-reported ADHD symptoms could be as disabling as ADHD.

The Biochemical and Genetic Basis for the Biosynthesis of Bioactive Compounds in Hypericum Perforatum L., One of the Largest Medicinal Crops in Europe.

Hypericum perforatum L. commonly known as Saint John's Wort (SJW), is an important medicinal plant that has been used for more than 2000 years. Although H. perforatum produces several bioactive compounds, its importance is mainly linked to two molecules highly relevant for the pharmaceutical industry: the prenylated phloroglucinol hyperforin and the naphtodianthrone hypericin. The first functions as a natural antidepressant while the second is regarded as a powerful anticancer drug and as a useful compound for the treatment of Alzheimer's disease. While the antidepressant activity of SJW extracts motivate a multi-billion dollar industry around the world, the scientific interest centers around the biosynthetic pathways of hyperforin and hypericin and their medical applications. Here, we focus on what is known about these processes and evaluate the possibilities of combining state of the art omics, genome editing, and synthetic biology to unlock applications that would be of great value for the pharmaceutical and medical industries.

Conformational distribution and ultrafast base dynamics of leadzyme.

The dynamic nature of ribozymes represents a significant challenge in elucidating their structure-dynamics-function relationship. Here, using femtosecond time-resolved spectroscopy and other biophysical tools, we demonstrate that the active site of leadzyme does not have a unique structure, but rather samples an ensemble of conformations that undergo picosecond structural changes. Various base modifications have a profound context-dependent impact on the catalysis.

Accuracy of Detection and Grading of Diabetic Retinopathy and Diabetic Macular Edema Using Teleretinal Screening.

PURPOSE: To determine the accuracy of a county teleretinal screening program of detecting referable diabetic retinopathy (DR) and treatable diabetic macular edema (DME), as well as to evaluate patient compliance with clinic follow-up after referral from teleretinal screening. DESIGN: Retrospective observational study. PARTICIPANTS: Patients in the Harris Health System (HHS, Houston, TX) older than 18 years of age who underwent teleretinal screening between July 2014 and July 2016. METHODS: Teleretinal imaging (TRI) consisting of single-field 45-degree nonmydriatic color fundus photography with referral thresholds of severe nonproliferative DR, proliferative DR, and significant DME. Teleretinal imaging results for all referred subjects were obtained and cross-referenced with dilated fundus examination findings with regard to DR severity and the presence of DME. Follow-up status was also noted. Subjects underwent OCT if deemed necessary by the examining specialist. Agreement between TRI and dilated fundus examination (DFE) findings was determined by calculating the Cohen κ coefficient. MAIN OUTCOME MEASURES: The primary outcome measure is agreement between TRI results and DFE findings with regard to DR severity and the presence of DME. The secondary outcome measure is compliance with follow-up. RESULTS: Of 1767 patients who were screened and referred for clinical examination, 935 (52.9%) attended their clinic appointment. Overall agreement between DFE and TRI was moderate (weighted κ 0.45) in terms of DR severity. There was agreement within one DR severity level in 86.2% of patients. The positive predictive value for detecting referable disease was 71.3%. Of patients referred for DME, 30.4% were deemed to have treatable DME. CONCLUSIONS: The HHS teleretinal screening program demonstrates a high level of accuracy in the detection and classification of referable DR, but a lesser degree of accuracy in the detection of treatable DME. Only slightly more than half of participants were compliant with follow-up after a TRI referral. This large-scale study provides insight into the utility of teleretinal screening in a county health care system.

First report of evaluation of K-M media: a new corneal preservation medium.

PURPOSE: To analyze the outcome of keratoplasty performed using Kalevar-Majumdar (K-M) media, a new synthetic viscous medium for preservation of the cornea. MATERIALS AND METHODS: The K-M media-preserved donor eye balls were kept in a bottle in a refrigerator at 4 degrees C till the corneas were used. Forty-eight consecutive keratoplasty cases of pseudophakic bullous keratopathy with vision less than counting fingers at one meter and operated by a single surgeon have been analyzed. Corneal donor button of 7.5 mm was used on the 7.0 mm recipient bed in all cases. Surgery was done with a standard technique. All the cases were examined daily for the first week and at the end of one month for graft clarity, epithelial defect and stromal edema. RESULTS: The K-M media-preserved corneal grafts remained clear at the end of the first week in 95.8% (46 of 48) cases and at the end of one month in 93.7% (45 of 48) cases. Donor epithelial haze cleared in 24 h in all cases. The stromal edema got cleared in the majority (91.7%, 44 of 48) within 24 h. Epithelial defect was seen in only 10.4% (five cases). There was no primary graft failure. CONCLUSION: K-M medium, a new viscous, synthetic corneal preservation medium, is a safe (no primary donor failure) alternative to conventional liquid corneal preservation media. K-M media-preserved eyes appear to have better preserved corneal epithelium with faster achievement of graft clarity postoperatively.

Comparison of Candida species isolated from children with and without early childhood caries: A descriptive cross-sectional study.

BACKGROUND: Early childhood caries (ECC) is characterized by the presence of one or more decayed, missing (due to caries), or filled teeth surfaces in any primary tooth, in a child below 6 years of age. Although ECC is primarily associated with high levels of maternal Streptococcus mutans, there has been an increased interest in finding the relationship between oral fungal flora and dental caries. OBJECTIVE: The aim of the study is to identify and characterize the Candida species and to compare the candidal isolates in children with ECC and without ECC. MATERIALS AND METHODS: The study was conducted on children below 6 years of age, who were categorized into ECC and non-ECC groups of fifty children each. Samples were collected using sterile cotton swabs and were inoculated on Sabouraud's Dextrose Agar and incubated at 37°C for 24 h. Candidal colonies were isolated, species identified and virulence factors tested for both ECC and non-ECC groups. RESULTS: The candidal carriage among the ECC children was found to be 84%, which was significantly higher than the non-ECC group of 24%. Candida albicans and non-albicans Candida (NAC) were isolated in both ECC and non-ECC groups. Phospholipase production was significantly high in ECC group whereas hemolysin production and germ tube formation showed no significant difference between the two groups. CONCLUSION: A significant correlation was found between the presence of Candida and ECC. NAC also plays an important role in the development of ECC. The virulence factors such as phospholipase may be responsible for the pathogenicity of Candida in the development of ECC.

Prevalence and Correlates of Psychological Distress in Adolescent Students from India.

OBJECTIVE: There are limited data on the prevalence and correlates of psychological distress among adolescents in India. This study assessed psychological distress among adolescents who attended school in Kerala, India. METHODS: A total of 7560 students from 73 schools, aged 12 to 19 years completed a self-administered questionnaire that included Kessler Psychological Distress Scale and other standardised instruments to assess various domains. RESULTS: Mild psychological distress was reported by 10.5%, moderate distress by 5.4%, and severe distress by 4.9% of students. Older age, not living with both parents, and urban residence were significantly associated with psychological distress (p < 0.05). Students who experienced psychological distress had a higher risk of reporting academic failure, alcohol and tobacco use, suicidality, and sexual abuse. Increasing severity of psychological distress was associated with higher odds of these correlates. CONCLUSIONS: Psychological distress is common among adolescents and its correlates with negative outcomes suggest the need for early recognition and treatment.

An audit of in-hospital cardiopulmonary resuscitation in a teaching hospital in Saudi Arabia: A retrospective study.

OBJECTIVES: Data reflecting cardiopulmonary resuscitation (CPR) efforts in Saudi Arabia are limited. In this study, we analyzed the characteristics, and estimated the outcome, of in-hospital CPR in a teaching hospital in Saudi Arabia over 4 years. METHODS: A retrospective, observational study was conducted between January 2009 and December 2012 and included 4361 patients with sudden cardiopulmonary arrest. Resuscitation forms were reviewed. Demographic data, resuscitation characteristics, and survival outcomes were recorded. RESULTS: The mean ± standard deviation age of arrested patient was 40 ± 31 years. The immediate survival rate was 64%, 43% at 24 h, and 30% at discharge. The death rate was 70%. Respiratory type of arrest, time and place of arrest, short duration of arrest, witnessed arrest, the use of epinephrine and atropine boluses, and shockable arrhythmias were associated with higher 24-h survival rates. A low survival rate was found among patients with cardiac types of arrest, and those with a longer duration of arrest, pulseless electrical activity, and asystole. Comorbidities were present in 3786 patients with cardiac arrest and contributed to a poor survival rate (P < 0.001). CONCLUSIONS: The study confirms the findings of previously published studies in highly developed countries and provides some reflection on the practice of resuscitation in Saudi Arabia.

Encoding of contextual fear memory requires de novo proteins in the prelimbic cortex.

BACKGROUND: Despite our understanding of the significance of the prefrontal cortex in the consolidation of long-term memories (LTM), its role in the encoding of LTM remains elusive. Here we investigated the role of new protein synthesis in the mouse medial prefrontal cortex (mPFC) in encoding contextual fear memory. METHODS: Because a change in the association of mRNAs to polyribosomes is an indicator of new protein synthesis, we assessed the changes in polyribosome-associated mRNAs in the mPFC following contextual fear conditioning (CFC) in the mouse. Differential gene expression in mPFC was identified by polyribosome profiling (n = 18). The role of new protein synthesis in mPFC was determined by focal inhibition of protein synthesis (n = 131) and by intra-prelimbic cortex manipulation (n = 56) of Homer 3, a candidate identified from polyribosome profiling. RESULTS: We identified several mRNAs that are differentially and temporally recruited to polyribosomes in the mPFC following CFC. Inhibition of protein synthesis in the prelimbic (PL), but not in the anterior cingulate cortex (ACC) region of the mPFC immediately after CFC disrupted encoding of contextual fear memory. Intriguingly, inhibition of new protein synthesis in the PL 6 hours after CFC did not impair encoding. Furthermore, expression of Homer 3, an mRNA enriched in polyribosomes following CFC, in the PL constrained encoding of contextual fear memory. CONCLUSIONS: Our studies identify several molecular substrates of new protein synthesis in the mPFC and establish that encoding of contextual fear memories require new protein synthesis in PL subregion of mPFC.

Prevalence and correlates of alcohol use among adolescents attending school in Kerala, India.

INTRODUCTION AND AIMS: Concern around potentially increasing alcohol use among young people has been growing in public discourse in India. However, there are few published studies on this issue. We studied the prevalence, patterns and correlates of alcohol use among adolescents in Ernakulam, Kerala State, India. DESIGN AND METHODS: A total of 7560 students in the age group 12-19 years from 73 schools completed a self-administered questionnaire incorporating standardised instruments to assess alcohol use. RESULTS: The overall prevalence of lifetime alcohol among adolescents use was 15% (23.2% among boys and 6.5% among girls) with prevalence increasing with age, and 25.3% of drinkers reported hazardous alcohol use. The mean age at onset of alcohol use was 13.6 years. Initiation of alcohol use typically preceded use of tobacco and illicit drugs. Most students reported initiation into alcohol use by family members in the context of family celebrations. The prevalence of alcohol use was higher among students from urban areas and those with a part-time job. Lower use was seen among Muslims. Lifetime alcohol use was associated with significantly higher tobacco and illicit drug use, suicidal thoughts, attention deficit hyperactivity disorder symptom-scores, history of non-contact sexual abuse and with poor academic performance. DISCUSSION AND CONCLUSIONS: Alcohol use among adolescents in India deserves greater attention than it has previously received, marked as it is by an early onset and associated with a range of negative mental health problems. [ Jaisoorya T S, Beena K V, Beena M, Ellangovan K, Jose D C, Thennarasu K, Benegal V. Prevalence and correlates of alcohol use among adolescents attending school in Kerala, India. Drug Alcohol Rev 2016;35:523-529].

NSP-Dependent Simple Nitrile Formation Dominates upon Breakdown of Major Aliphatic Glucosinolates in Roots, Seeds, and Seedlings of Arabidopsis thaliana Columbia-0.

One of the best-studied plant defense systems, the glucosinolate-myrosinase system of the Brassicales, is composed of thioglucosides known as glucosinolates and their hydrolytic enzymes, the myrosinases. Tissue disruption brings these components together, and bioactive products are formed as a consequence of myrosinase-catalyzed glucosinolate hydrolysis. Among these products, isothiocyanates have attracted most interest as chemical plant defenses against herbivores and pathogens and health-promoting compounds in the human diet. Previous research has identified specifier proteins whose presence results in the formation of alternative product types, e.g., nitriles, at the expense of isothiocyanates. The biological roles of specifier proteins and alternative breakdown products are poorly understood. Here, we assessed glucosinolate breakdown product profiles obtained upon maceration of roots, seedlings and seeds of Arabidopsis thaliana Columbia-0. We identified simple nitriles as the predominant breakdown products of the major endogenous aliphatic glucosinolates in root, seed, and seedling homogenates. In agreement with this finding, genes encoding nitrile-specifier proteins (NSPs) are expressed in roots, seeds, and seedlings. Analysis of glucosinolate breakdown in mutants with T-DNA insertions in any of the five NSP genes demonstrated, that simple nitrile formation upon tissue disruption depended almost entirely on NSP2 in seeds and mainly on NSP1 in seedlings. In roots, about 70-80% of the nitrile-forming activity was due to NSP1 and NSP3. Thus, glucosinolate breakdown product profiles are organ-specifically regulated in A. thaliana Col-0, and high proportions of simple nitriles are formed in some parts of the plant. This should be considered in future studies on biological roles of the glucosinolate-myrosinase system.

Improving quality for maternal care - a case study from Kerala, India.

BACKGROUND: The implementation of maternal health guidelines remains unsatisfactory, even for simple, well established interventions. In settings where most births occur in health facilities, as is the case in Kerala, India, preventing maternal mortality is linked to quality of care improvements. CONTEXT: Evidence-informed quality standards (QS), including quality statements and measurable structure and process indicators, are one innovative way of tackling the guideline implementation gap. Having adopted a zero tolerance policy to maternal deaths, the Government of Kerala worked in partnership with the Kerala Federation of Obstetricians & Gynaecologists (KFOG) and NICE International to select the clinical topic, develop and initiate implementation of the first clinical QS for reducing maternal mortality in the state. Description of practice: The NICE QS development framework was adapted to the Kerala context, with local ownership being a key principle. Locally generated evidence identified post-partum haemorrhage as the leading cause of maternal death, and as the key priority for the QS. A multidisciplinary group (including policy-makers, gynaecologists and obstetricians, nurses and administrators) was established. Multi-stakeholder workshops convened by the group ensured that the statements, derived from global and local guidelines, and their corresponding indicators were relevant and acceptable to clinicians and policy-makers in Kerala. Furthermore, it helped identify practical methods for implementing the standards and monitoring outcomes. LESSONS LEARNED: An independent evaluation of the project highlighted the equal importance of a strong evidence-base and an inclusive development process. There is no one-size-fits-all process for QS development; a principle-based approach might be a better guide for countries to adapt global evidence to their local context.