Several controversial findings have been reported on treatment outcomes for diabetic stroke patients that received thrombolysis therapy in the hospital. We determined whether the association between telestroke technology, thrombolysis therapy and clinical risk factors in diabetic acute ischemic stroke may result in the inclusion or exclusion or more diabetic ischemic stroke patients for thrombolysis therapy.Retrospective data that comprises of a total of 3202 acute ischemic stroke patients from a regional stroke registry that contained telestroke and non telestroke patients with a primary diagnosis of acute ischemic stroke of which 312 were identified as diabetic stroke patients were used in this study. Multivariate logistic regression models were used to determine the associated pre-clinical risk factors, and demographics associated with recombinant tissue plasminogen activator (rtPA) therapy in a subset of diabetic acute ischemic stroke patients in the telestroke and non-telestroke settings.In the telestroke, only higher International Normalized Ratio (INR)  was associated with exclusion from thrombolysis. Direct admission  and telestroke  were independent predictors in the inclusion for thrombolysis therapy. In the non telestroke, older age (> 80 years) , higher blood glucose level , higher INR , and renal insufficiency  were associated with exclusion while higher NIH stroke scale  was associated with inclusion for thrombolysis in the non telestroke.The non-telestroke setting admitted more diabetic stroke patients to the hospital, but more were excluded from thrombolysis therapy when compared with the telestroke setting. Measures to improve clinical risk factors that excluded more diabetic ischemic stroke patients in the non telestroke will improve the use of thrombolysis in the treatment of diabetic acute ischemic stroke patients. Diabetes mellitus is a frequently identified comorbid risk factor in acute ischemic stroke. The risk of ischemic stroke in diabetic patients is twofold higher when compared to people without diabetes . This unAlthough diabetes is not an absolute or relative exclusion criteria for thrombolysis, a low rate of thrombolysis therapy has been reported in diabetic ischemic stroke patients due to concerns over poorer outcomes . ProposeIt has been shown that a practice-based model of telestroke can manage pretreatment clinical risk factors for thrombolysis therapy relaxing the criteria for the inclusion or exclusion for thrombolysis in ischemic stroke patients . AlthougRetrospective data were collected from the acute ischemic stroke registry of Greenville Health System (GHS) between January 2010 and June 2016. The registry has been described in our previous studies . PatientUpon admission, all patients underwent brain computed tomography. Patients with subarachnoid and intracerebral hemorrhage were excluded in our analysis. A standardized stroke protocol was used in all patients, including T2-weighted, T1-weighted, and diffusion-weighted images. Data on symptom onset time and the admission to Emergency Department (ED) for both telestroke and non telestroke diabetic stroke patients were collected. Patients that were directly admitted to the ED or with emergency medical services (EMS) and those with indirect admission by being transferred to the ED in the telestroke or non telestroke from another hospital were also identified. Data on patient demographics, including age, sex, race, and ethnicity were also extracted Information on the ambulation status prior to event, during and at discharge were also collected. Ethical approval was obtained from the Institutional Review Board of Greenville Health System and the institutional Committee for Ethics.T test. All variables presented in Tables 2 and Classification Plots. Multicollinearity of variables were assessed with variance inflation factor analysis to confirm independence of variables included in regression model.The SPSS package version 20  for Windows was used for all statistical analyses and P < 0.05 was used to establish statistical significance for all comparisons between groups. We used univariate analysis to analyze baseline characteristics including age, gender, medical history, prestroke treatments and admission parameters such as blood glucose and stroke severity. This allowed us to determine baseline or pre-clinical risk factors that were associated with inclusion or exclusion for recombinant tissue plasminogen activator (rtPA). All discrete variables were represented as number (percentage) and comparisons between groups were made using Pearson’s Chi Squared analyses. Descriptive statistics were calculated for the demographic and clinical characteristics of patients. All continuous variables were presented as mean (STD), and comparisons between groups were determined using the Student’s A total of 3202 acute ischemic stroke patients were collected from the stroke registry, 312 were identified as diabetic stroke patients. Of the 312, 180 were in the non-telestroke setting and 132 in the telestroke setting. Comparisons between the baseline demographic and clinical characteristics of telestroke and non-telestroke diabetic acute ischemic stroke patients are presented in Table As shown in Table In a diabetic acute ischemic stroke population, patients that present with obesity, directly admitted to emergency department, and received thrombolysis therapy have higher odds of being associated with the telestroke setting. Following the adjustment for comorbidities, the telestroke setting represents the strongest predictor for the administration of thrombolysis therapy. In both telestroke and non-telestroke diabetic acute ischemic stroke patients, direct admission represents a predictor for administration of thrombolysis therapy. In the univariate analysis, non-telestroke diabetic stroke patients who received thrombolysis were more likely to be younger, have a lower blood glucose level, lower creatinine, lower INR, and present with a better ambulatory status at baseline than the patients who did not receive thrombolysis. In the telestroke setting, diabetic acute ischemic stroke patients who received thrombolysis were more likely to have a worse ambulatory status at presentation and more likely to be directly admitted to the emergency department.In the adjusted analysis for the total diabetic stroke population, only direct admission and being treated in the telestroke setting were independent variables associated with administration of thrombolysis therapy. The non-telestroke setting admitted more diabetic stroke patients, but more were excluded from thrombolysis therapy when compared with the telestroke setting. This may be connected with a higher rate of hospital admission of patients with highly variable clinical risk factors, resulting in the exclusion of more admitted patients from thrombolysis therapy when compared with the telestroke setting. In the adjusted analysis for the non-telestroke setting, age (> 80), higher blood glucose level, and renal insufficiency were all associated with exclusion from thrombolysis. The benefits of thrombolysis therapy have been shown in many studies , 22–30. A major finding in this study is that our multivariate model predicted a direct association of treatment in the telestroke setting as an independent variable with the highest odds for the inclusion of diabetic stroke patients for thrombolysis therapy. Moreover, following adjustment for baseline demographic and clinical risk factors in the telestroke network, only diabetic stroke patients with higher INR were excluded from thrombolysis, and the effect was not significant. These findings differ from the non-telestroke setting in which diabetic stroke patients with increased age, higher blood glucose level, higher INR, and renal insufficiency were all pre-clinical risk factors that predicted exclusion from thrombolysis therapy. The finding that in the non-telestroke setting, diabetic stroke patients with complicated pre-clinical risk factors were associated with a higher likelihood of exclusion from thrombolysis therapy, suggests a more stringent exclusion criteria when compared with the telestroke setting. Therefore, it is possible that telestroke technology provides a real-world clinical setting that streamlines in-hospital evaluation with less stringent exclusion criteria, allowing stroke neurologist to consult quickly on whether or not administer thrombolysis therapy. This may enable an increase in the rate of use and efficiency of the timeline for administration of thrombolysis in the treatment of diabetic acute ischemic stroke patients.There are limitations to our study. First, our study is limited by its retrospective design, although data was collected using an established prospective stroke registry, a risk of selection bias is possible. Furthermore, this is unicenter stroke registry and does not allow for the generalization of our findings. Moreover, information about the management of diabetes mellitus (type I or type II) was not included in our analysis. The relatively small groups of patients of diabetic stroke patients did not increase the power of our analysis. The strengths of our study are that in the non-telestroke setting, increased age, higher blood glucose level, renal insufficiency were pre-clinical risk factors that predicted the exclusion from thrombolysis therapy, while only INR predicted a non-significant exclusion from thrombolysis therapy in the telestroke setting. Our multivariate model was able to identify treatment in the telestroke setting as an independent variable with the highest prediction for the inclusion of diabetic stroke patients for thrombolysis therapy. Finally, we found that in older diabetic stroke patients (> 80 years), exclusion maybe linked with pre-treatment functional status that includes history of higher blood glucose level, higher INR, and renal insufficiency.Diabetes is not an exclusion criterion for thrombolysis, however, a low rate of thrombolysis therapy has been reported in diabetic acute ischemic stroke patients. More studies are necessary to determine how identified exclusion risk factors in the non-telestroke setting can be improved to provide a real-world clinical setting with less stringent exclusion criteria for thrombolysis therapy.


There is consistent evidence that caregiving for grandchildren is psychologically, emotionally and physically challenging for grandparents. However, many of them may not be able to receive the support they need from their own families or public services. Using the Selective Optimization with Compensation framework and principles of solution-oriented cognitive models, we implemented a six-week session pilot program to increase grandparents’ effective coping strategies, reduce stress, and improve parenting and communication with grandchildren. Preliminary data from 22 grandparents suggest that participants improved their use of effective selection strategies to set priorities and pursue goals related to their wellbeing and the quality of their relationship with grandchildren. Their pre-post test scores also indicated a higher level of parental efficacy, social relations and confidence in organizing their time and priorities. Results indicate that focusing on solution-oriented strategies may empower grandparents to improve their psychological adjustment and take care of their needs.


Models of G protein-coupled receptor (GPCR) signaling have dramatically altered over the past two decades. Indeed, GPCRs such as the follicle-stimulating hormone receptor (FSHR) have contributed to these new emerging models. We now understand that receptor signaling is highly organized at a spatial level, whereby signaling not only occurs from the plasma membrane but distinct intracellular compartments. Recent studies in the role of membrane trafficking and spatial organization of GPCR signaling in regulating gonadotropin hormone receptor activity has identified novel intracellular compartments, which are tightly linked with receptor signaling and reciprocally regulated by the cellular trafficking machinery. Understanding the impact of these cell biological mechanisms to physiology and pathophysiology is emerging for certain GPCRs. However, for FSHR, the potential impact in both health and disease and the therapeutic possibilities of these newly identified systems is currently unknown, but offers the potential to reassess prior strategies, or unveil novel opportunities, in targeting this receptor. The follicle-stimulating hormone receptor (FSHR) belongs to the superfamily of G protein-coupled receptors (GPCRs). With more than 800 members in humans they represent the largest family of signaling receptors and a major, successful, drug target . The canThis review will discuss our current understanding of the molecular mechanisms and signaling roles of membrane trafficking of FSHR, and how gonadotropin hormone receptors have shed light on novel cell biological pathways potentially applicable to many GPCRs. We will primarily focus on post-endocytic intracellular trafficking, and then discuss how this novel cell biology could shed light on specific facets of FSH/FSHR function and its implications to endocrine function.21/18 and the fully glycosylated FSH24     . This labinding) . While pFollowing internalization, GPCRs are trafficked to endosomes where they are sorted to either a plasma membrane recycling pathway, or to the lysosomal pathway for degradation. Such pathways program the temporal profile of G protein signaling, by regulating resensitization/hormone recovery (recycling) or permanent signal termination (degradation), Additionally, the sorting fate of a GPCR pharmacologically manipulated and altered in disease . HoweverThe textbook model of cargo sorting depicts the Rab5 early endosome (EE) as the common post-endocytic compartment from which receptors are first sorted to opposing fates. GPCRs sorted to a degradative pathway following internalization are trafficked from EEs to Rab7 positive late endosomes. Receptors are involuted into vesicles within the lumen of these endosomes, to form multivesicular bodies (MVBs). MVBs will then fuse with lysosomes resulting in protein degradation. GPCRs will engage with this pathway with distinct kinetics and for those receptors targeted to the recycling pathway, chronic ligand stimulation will reroute receptors to this degradative pathway as part of the mechanism of downregulation. Classically, lysosomal targeting of different receptors is via ubiquitination at lysine residues and engagement with endosomal sorting complex required for transport (ESCRT)-dependent degradation, however, GPCRs exhibit ubiquitin-independent and ESCRT-independence in their mechanisms of degradation . GPWhile the above describes that the recycling pathway of FSHR, and other GPCRs, involves a one-step mechanism with its recycling sequence and interacting partner, which may be unique to a given GPCR, we now know that both common and receptor-specific post-endocytic mechanisms exist. Furthermore, sequence-directed recycling  occur via a complex, multi-step system, with the GPCR's own signaling playing a key role in driving receptors to these distinct cellular fates, in addition to trafficking regulating the signal from that receptor. This interconnected property of post-endocytic trafficking and GPCR signaling has been highlighted recently through studies of the gonadotropin hormone receptors and will be discussed next.The internalization of GPCRs into the endocytic network is no longer viewed as a mechanism to only control plasma membrane signaling but to also provide additional platforms to continue or reactivate signaling from intracellular compartments, including G protein signaling. Furthermore, this spatial control of signaling has been shown to be important for cells to decode common second messenger signaling molecules, such as cAMP signaling, activated by many GPCRs, into specific downstream cellular functions –61. IntrMany GPCRs, including the human FSHR and LHR undergo the regulated recycling pathway described in Section Post-endocytic intracellular trafficking pathways of FSHR. Our studies first identifying the VEE with the gonadotropin hormone receptors were initially driven as way to understand why GPCRs have distinct recycling sequences if there is a common primary function, i.e., sorting to the recycling pathway. Thus, comparisons were first made between the β2AR and LHR that initially seem quite similar in their signaling and trafficking profiles . Both ar3 production and calcium release  to possibly distinct functional consequences. It also raises the intriguing possibility that GPCR activity, can be reprogrammed by the cell to alter the compartment the receptor is targeted to via altering expression of key proteins in this pathway, e.g., GIPC, APPL1 Figure . Such a These recent advances in FSHR function from its distinct roles in extragonadal tissues and novel cell biological functions demonstrates that FSHR is a good example of a GPCR that achieves signal diversification via multiple strategies and a prototype receptor for understanding novel facets of GPCR function. Whether such pathways are then perturbed in disease and if we can harness these properties of GPCRs therapeutically needs to be addressed.While much has been uncovered about novel intracellular trafficking systems, there are numerous outstanding questions at the molecular level. A major one to be tackled in the future is the identifying the molecular composition (both protein and lipid content of the membrane) of the VEEs, as so far APPL1 is the only known protein to reside there and is only present in ~50% of the VEE population . While nThe ability to pharmacologically exploit these new GPCR signaling models and target the intracellular signaling receptor specifically has been recently demonstrated for three distinct GPCRs. Employing cholestenol-conjugated antagonists, which accumulate in the lumen of endosomal compartments, can block endosomal signaling from the neurokinin 1 receptor and calcitonin-gene related peptide receptor and has been shown to be an effective nociceptive target in animal models , 97. Whiin vivo. This would provide the opportunity to be able to target these intracellular signaling modalities with high precision, in order to create the next generation of therapeutics for reproductive medicine.In summary, our understanding of the complexity of GPCR signaling pathways via the tight control of their intracellular location has been advanced through studies on the gonadotropin hormone receptors. Such mechanisms have highlighted the interconnected nature of these intracellular systems, and thus a primary future goal is to further understand the significance of these molecular systems to health and disease if they are to be of therapeutic value. The critical nature of intracellular sorting of FSHR to signaling has been demonstrated, so it is not a question of is it important, but rather how the intricacies of modulating receptor from one intracellular compartment to another impact specific functions All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


An extensive body of theoretical work has advocated the use of multiple human wellbeing indicators to assess the outcomes of agricultural investments in Sub-Sahara Africa (SSA). However, few studies have actually achieved it. This study investigates the human wellbeing outcomes of involvement in industrial crop production in Ghana by comparing the levels of different objective and subjective wellbeing measures for groups involved in industrial crop production as plantation workers and smallholders, and groups not involved (i.e. control groups). We use household income, adult consumption and the multidimensional poverty index (MPI) as indicators of objective wellbeing. We measure subjective wellbeing through self-reported levels of satisfaction with life, worthwhileness, happiness and anxiousness. Propensity Score Matching (PSM) analysis is used to assess whether involvement in industrial crop production increases household income and consumption. Overall, for most indicators of objective wellbeing industrial crop outgrowers, smallholders and independent smallholders are better off compared to other groups in their respective sites (in terms of mean scores), but involvement does not necessarily brings human wellbeing benefits . On the other hand plantation workers are either worse off or have similar level of objective human wellbeing with control groups in their respective sites (in terms of mean scores), but involvement sometimes brings human wellbeing benefits . However, workers tend to benefit from access to plantation infrastructure, which has a positive effect to their multi-dimensional poverty. In most cases the objective wellbeing measures do not correlate well with self-reported levels of subjective wellbeing. It is important to combine such indicators when evaluating the human wellbeing outcomes of agricultural investments in order to obtain a more comprehensive outlook of whether industrial crop production can become a valuable rural development strategy in SSA. Industrial crop production has been expanding in several parts of Sub-Sahara Africa (SSA) during the past decades . PerhapsHowever, the human wellbeing and rural development outcomes of engaging in the production of such crops depends on various factors such as the crop, mode of production , available markets, and the local socioeconomic and environmental context –14. For In any case, it is important to assess the household-level human wellbeing outcomes for the different possible types of involvement in industrial crop value chains  in order to understand their possible rural development and poverty alleviation benefits in SSA . HoweverThe aim of this study is to undertake a comprehensive assessment of the objective and subjective human wellbeing outcomes of involvement in the production of different industrial crops (under various production models) in Ghana. We use multiple subjective and objective measures to gain a comprehensive understanding of how involvement in such crop systems can affect the human wellbeing at the household level. We focus on crops that can potentially be used as biofuel feedstocks , as such crops have experienced a substantial expansion in the country. Jatropha was the most promoted industrial crop in Ghana  before iTo achieve this, we compare households involved in industrial crop production  with households not involved (i.e. control groups). We focus on three operational industrial crop projects in Ghana, namely sugarcane , jatropha (plantation-based production) and oil palm . We employ various indicators of objective and subjective wellbeing. In particular, we use income, consumption and the multidimensional poverty index as measures of objective wellbeing ,35–38, aFirst we outline the adopted methodology, including the study sites, and the data collection and analysis methods (see “Methodology”). Subsequently we report the findings for the different indicators in each site, as well as their correlations .The selected study sites have very different characteristics. First, they represent the main different biofuel feedstock options in Ghana  . BetweenTragelaphus spekii) under the International Union for Conservation of Nature (IUCN) Red List. In 2012, with the intention to protect the landscape and the Sitaunga species, the 15 local communities located within the catchment area formed the Avu Lagoon Community Protected Area .,68.35,68q denotes the number of people identified as poor and n the total number of people in the sampleiC(k) denotes the censored deprivation score which (k) indicates the share of possible deprivation experienced by the poor person i.The Adjusted Headcount Ratio (Mo), is determined based on Eq ,Mo=H×A(mple see . and “A”mple see .H=q/n(7Considering the Eqs –8), the, the8), Several studies have already reported the detailed steps for using the MPI in terms of whether the household is deprived in a specific indicator (deprivation focused), the number of indicator dimensions (poverty-focused) and ignoring of households not meeting the dual cut-offs in MPI calculations. The reader interested in the methodology of the MPI is referred to ,64,68,69To explore whether groups involved in industrial crop production were significantly better off than the control groups, we used the bootstrap resampling approach for statistical inference of multi-dimensional poverty between groups . We alsoAs outlined above, the MPI calculation procedure aggregates the deprivation scores of poor households into a single index, which is obtained by multiplying the multidimensional poverty headcount ratio and the intensity of poverty. This makes it impossible to obtain individual MPIs for each household, as for other indicators of objective wellbeing . Therefore it is impossible to undertake PSM analysis using the MPI as an outcome variable, as performed for the other objective wellbeing indicators (see “Income and expenditure”). This means that caution needs to be paid when using the results of this study to understand whether there is a causal relationship between involvement in industrial crop production is an agent of poverty alleviation in terms of MPI.Several studies have made the point that measures of subjective wellbeing (SWB) need to be used alongside measures of objective wellbeing –25. In o(i) Overall, how satisfied are you with your life nowadays?(ii) Overall, how happy did you feel yesterday?(iii) Overall, how anxious did you feel yesterday?(iv) Overall, how worthwhile are the things that you do in your life?U test to assess whether the self-reported levels for each indicator were statistically significant between groups.We used a 4-level Likert scale to assess each indicator . We then used descriptive statistics based on the distribution of the responses for each dimension –77. SubsWe do not undertake the PSM analysis for the different indicators of subjective wellbeing. First of all, subjective wellbeing can be affected by many factors that go beyond household livelihoods  such various cultural, socioeconomic, environmental and personal factors . FurtherThe development of the research protocol was informed by a similar protocol that elicited the impact of industrial crops in southern Africa , and adoTo ensure the smooth and proper community entry, prior to contacting respondents we undertook meetings with traditional authorities and representatives of the private companies and farmers’ associations operating in each study site. This was necessary as local chiefs are influential members of the local communities in Ghana. Furthermore, we sought the permission of the two industrial crop companies in Kwae (GOPDC) and Yeji (Smart Oil) to avoid causing problems to the interviewed workers. Eventually we were allowed to interview plantation workers in company estates , which wWe sought oral informed consent from all respondents before each interview, explaining the purpose and procedures of the research. Oral consent was considered as the most appropriate approach considering the high illiteracy rates in the local communities. Through this process seeking oral informed consent we ensured that all respondents were fully aware what their participation in this study entailed, and explained any concerns related to anonymity and disclosure of personal information. We did not interview any minors. The age ranges of the respondents in each site are: Dabala: 24–73 years; Yeji: 21–90 years; Kwae: 20–75 years.Participation in the household survey was voluntary. Respondents reserved the right to decline being interviewed, and were informed that possible refusal would not lead to any adverse consequences. No payments were made to participants and all surveys were anonymised to avoid the identification of respondents.n = 100) having at least some level of formal education (i.e. at least basic education).Tables Overall the smallest household sizes are reported in Kwae , with GOPDC workers having the smallest households, largely due to the fact that most originate from other parts of Ghana. Worker households usually consist of adult members with few children. With the exception on workers, there are no other significant differences in terms of household sizes and composition between the other study groups . Similarly, there is no significant difference in total household size and composition between sugarcane smallholders and the control group in Dabala. On the contrary there are significant differences in household size and composition between permanent jatroph workers (YJ_JPW) and the control group in Yeji. Overall, the jatropha site exhibits the largest mean household sizes compared to other sites, at 5.7, 5.8 and 6.3 persons per household for permanent workers, seasonal workers and the control group respectively. At the same time the study groups in Yeji have the highest fraction of children to adult members among all sites. These patterns reflect well the context of Ghana, where households in the savanna ecological zone (where Yeji is located) have larger household sizes and higher fertility rates compared to other parts of the country ,80.When it comes to land ownership there is no significant difference in total land size and cropland between sugarcane smallholders and control groups in Dabala Tables  and 6. IIn the Dabala site (sugarcane), sugarcane growers report significantly higher mean income levels compared to the control group . FurtherIn Yeji (jatropha), control group respondents report substantially higher mean income compared to permanent and seasonal workers . ActuallIn Kwae , mean income levels are very significantly different between all groups, with the exception of outgrowers (KW_OG) and independent smallholders (KW_ID) . Oil palWe use the PSM technique to assess whether there is a causal relationship between involvement in industrial crop production and improvement in the four objective wellbeing indicators. The PSM analysis for sugarcane (DB_S) and non-sugarcane (DB_NS) producers uses the radius caliper (0.1) matching algorithm and suggests that engagement in sugarcane production leads to both significantly higher total household income  and per capita household income  . SimilarThe PSM estimation for permanent jatropha workers (YJ_JPW) and the control group (YJ_JC) suggests that engagement in permanent jatropha employment leads to higher total income (by GHC 500.78) and per capita income (by GHC146.95), but this is not statistically significant . To achiThe PSM analysis in Kwae suggests that involvement in oil palm production as workers (KW_W), outgrowers (KW_OG) and independent smallholders (KW_ID) leads to declining levels of most objective wellbeing indicators . The PSMIn summary, all groups engaged in industrial crop production  exhibit very different poverty levels compared to their respective control groups. Groups involved in sugarcane and oil palm production are better off than their respective control groups, while the opposite is true for jatropha workers. In fact, among all groups seasonal workers in the jatropha plantation (YJ_JSW) have the highest MPI (0.400). To check the robustness of the MPI results, we change indicator weights . Table 1In Dabala the patterns of deprivation across the different MPI dimensions is fairly similar for sugarcane smallholders and the control group, with both groups exhibiting high deprivation for access to clean energy and improved sanitation.In Yeji, the patterns of deprivation are almost identical between groups for the living standards dimension. Overall, while it is not clear-cut, the control group tends to have the lowest deprivation scores for most indicators. It is also worth mentioning that compared to all other sites, groups in Yeji have the highest deprivation for education, largely due to the fact that some of the sampled communities do not have basic school facilities in their vicinity.Finally, in Kwae , deprivation patterns are quite mixed between groups. One interesting finding is that GOPDC workers have low deprivation for access to clean drinking water, sanitation, and clean cooking fuel as the company has different facilities within the estate for social service delivery such as schools, pipe water supply and hospital. For most other indicators oil palm outgrowers (KW_OG) and independent smallholders (KW_ID), have lower deprivation compared to GOPDC workers (KW_GW) and the control group (KW_C).U, However, differences of the mean scores are not always statistically significant between groups  correlation between consumption, and two indicators of subjective wellbeing  for permanent jatropha workers , sugarcane smallholders have lower levels of multi-dimensional poverty, and higher levels of income and consumption than the control group . This is consistent with recent studies in other SSA countries that have reported relatively higher income levels and lower poverty levels for sugarcane smallholders ,81–83. WHowever, the higher levels of objective wellbeing of sugarcane growers do not necessarily translate in higher levels of subjective wellbeing. Indeed most sugarcane smallholders have reported low levels of satisfaction with life, worthwhileness, and happiness , Fig 4. In Yeji (jatropha), the control group has lower poverty (in terms of MPI) and higher mean income than jatropha workers. The relatively higher poverty of jatropha workers might reflect the fact that by the time of the survey they were employed only for 2–5 years, which is possibly a short period of time for strong poverty alleviation effects to materialize, especially those related to some of the longer-term MPI components . However, the PSM analysis suggests that engagement in jatropha production increases income for permanent workers, which means that such longer-term MPI indicators might improve in the future. Our results contrast a recent study that found lower levels of multi-dimensional poverty for permanent jatropha plantation workers in Mozambique , but conThe relatively lower human wellbeing levels of jatropha workers is also reflected in the subjective wellbeing indicators, as a notable proportion of control group respondents reported higher satisfaction with life, happiness and worthwhileness compared to jatropha workers , Fig 4. In Kwae , we observed lower MPI levels for GOPDC workers compared to other groups , despite their relatively lower mean income amongst study groups (see “Income and expenditure”). This discrepancy is largely due to the fact that their employment entitles them to some social services offered by GOPDC such as access to piped water, electricity, a hospital and a school. Similar findings have been reported for workers in other types of plantations in Mozambique (jatropha), and Malawi/Swaziland (sugarcane) .Our study also confirms other studies in Ghana (and elsewhere in SSA), which identify oil palm income as a major livelihood source for smallholders and outgrowers –86. HoweFirst, due to its size and spacing requirement, oil palms need extensive land areas in order to generate large economic output . Indeed oil palm producers have on average much larger plots compared to control groups and workers Tables  and 6. GSecond, market prices can have a strong effect on the income obtained from oil palm production. Even though the GOPDC is the main oil palm market option in Kwae, there are multiple other informal buyers operating in the area (see “Study sites”). Outgrowers are contractually obliged to sell their harvest to the GOPDC, but independent smallholders can decide where to sell according to price signals. Generally speaking, GOPDC offers a slightly lower price for fresh fruit bunches, compared to informal buyers (respectively GHC 480 and 520 per tone of fresh bunches during the time of survey). This ability to tap into better market options might explain the relatively higher mean income  and moreAs outlined in the “Introduction” and “Methodology”, the main group comparison in this paper are performed for groups within the same site. This is because it is difficult to compare and generalize results between sites, due to the pre-existing differences and the specific socioeconomic/environmental contexts in each site. However, we can identify three interesting patterns between sites:levels of objective wellbeing decline when moving on a wet-dry gradient (mean scores);industrial crop outgrowers, smallholders and independent smallholders are generally better off than other groups in their respective sites (mean scores), but involvement in industrial crop production does not necessarily bring human wellbeing benefits ;plantation workers are either worse off or have similar level of human wellbeing with control groups in their respective sites (mean scores), but engagement in plantation work sometimes brings human wellbeing benefits .Regarding (a), mean incomes in the oil palm area (Kwae) are generally higher than in the sugarcane  and jatropha area (Yeji), both for groups involved in industrial crop production and control groups . In partRegarding (b) and (c), when looking at the mean income and consumption levels, the outgrowers and smallholders are better off than other study groups in each site, while plantation workers are either worse off or have the same levels compared to their respective control groups Tables  and 8. HHowever, it is worth noting that it is highly possible that these multidimensional poverty benefits are precarious. For example, the access to social services might cease (taking a toll on MPI scores), once plantation employment is lost either through termination or company collapse . This isAs already discussed above there is good correlation between objective wellbeing indicators such as income and consumption (see “Correlations between indicators of objective and subjective wellbeing”). However, the levels of these indicators do not always correspond with MPI, due to the fact that some of the social benefits that plantation workers receive affect “disproportionally” overall MPI levels .There are only a few strong correlations between indicators of objective and subjective wellbeing (see “Correlations between indicators of objective and subjective wellbeing”), but it is not clear from our cross-sectional data why this happens. In theory larger income can help households attain greater material welfare thereby contributing positively to subjective wellbeing –26. HoweAt the group level, relatively high incomes with low correlation with subjective wellbeing indicators might indicate "frustrated achievers" . In our When it comes to job security, plantation employment is among the few formal employment opportunities in poor rural areas of Ghana . HoweverSimilarly, most industrial crop smallholders rely on loans to purchase agricultural inputs such as fertilizers and pesticides. FGDs have suggested that most smallholders enter loan agreements before knowing the actual crop price, as this is established post-harvest depending on many different factors. In fact there is a significant risk of defaulting loan repayments, especially considering yield variability, price volatility, market access , and climatic variability  (see previous section). Many FGD respondents actually suggested that such factors could cause substantial stress to many famers, whose families depend on industrial crops, possibly influencing their self-reported levels of subjective wellbeing.Finally, the general lack of strong correlation between objective and subjective wellbeing measures suggests that the different indicators can indeed shed light to different aspects of human wellbeing. Thus, studies that seek to provide a comprehensive understanding of whether different agricultural interventions can improve human wellbeing should contain a combination of different human wellbeing indicators.Our study identifies two important aspects that need to be considered in industrial crop policies and practices in Ghana, and elsewhere in SSA: (a) securing stable markets for smallholders; (b) improving plantation performance for workers and surrounding communities.First, stable markets are a strong pre-condition that can allow smallholders reap the benefits of involvement in industrial crop production. This aspect came out strongly in the open-ended survey questions and FGDs in Kwae and Dabala, and is reflected in some of the objective and subjective wellbeing indicator patterns (see previous sections). As already discussed, there are multiple market options in Kwae for oil palm, which allows independent smallholders to choose the most competitive option for selling their produce. This is possibly reflected in their better performance compared to the outgrowers, in terms of higher mean income and better comparison to the control group in the PSM analysis (see “Patterns between groups in each site”), whose only market option is GOPDC. However, the opposite is observed in Dabala, where sugarcane growers often indicate that the lack of stable markets is a key risk of the engagement in sugarcane production. With a pretty much collapsed sugarcane sector in Ghana, stable market development should become a key priority in future efforts to revamp sugarcane production in the country. The government adopted recently a sugarcane policy in its efforts to boost the sector , includiSecond, commercial agriculture has been promoted in Ghana during the past decade mainly through plantation-based large-scale land acquisitions for jatropha production . DespiteIn this context, some of the possible measures to ensure the delivery of broader sustainability benefits from industrial crop plantations could be: (a) to improve the working conditions of plantation workers, and (b) ensure that the wider community benefits from the infrastructure and social services developed by plantations, even following company restructuring or collapse. In particular, the adoption of sustainability standards can possibly catalyze: (a) improvements in income levels and broader working conditions for plantation workers, (b) development of social services . GOPDC aFinally, despite the rich information generated presented in the previous sections, establishing causality has been a major challenge. In other words it has been very challenging to ascertain whether those households that are better off in terms of income, consumption and MPI got better off due to their involvement in industrial crop production, and were not already better off. The inability to adopt a random sampling approach and the lack of a baseline study with longitudinal data curtails our ability to definitively conclude this with certainty ,104. To This study investigated the human wellbeing outcomes of involvement in industrial crop production using evidence from multiple sites, and objective and subjective wellbeing measures. In general, the results suggest that sugarcane and oil palm growers are better off compared to their respective control groups in terms of lower multidimensional poverty, and higher mean income and consumption. However, the PSM analysis suggests that engagement in industrial crop cultivation might not actually deliver positive human wellbeing outcomes in the oil palm study site . On the other hand, results for workers in oil palm and jatropha plantations are mixed. Workers have consistently lower (or similar) mean income and consumption than their respective control groups. However, the PSM analysis suggests that employment in the jatropha plantation leads to higher income, while employment in oil palm plantations has the exactly opposite effect. Interestingly oil palm workers have the lowest levels of multidimensional poverty than any other groups in Kwae, which possibly reflects their access to the social services offered by the GOPDC.It is particularly interesting to point out that for most groups there is no strong correlation between the different measures of objective and subjective wellbeing. It is highly possible that diverse factors such as job/market insecurity, climate variability (e.g. floods) and self-comparisons with neighbors could influence the levels of reported subjective wellbeing. This suggests that the different metrics used in this study capture indeed different aspects of human wellbeing. It is therefore important to combine such metrics in order to obtain a more holistic overview of the performance of agricultural investments in SSA. Such a multi-dimensional approach could offer decision-makers and practitioners with more insights as to the potential of different rural development interventions in the continent.S1 File(DOCX)Click here for additional data file.S2 File(DOCX)Click here for additional data file.S3 File(XLSX)Click here for additional data file.


Substantial pressure to reduce antibiotic use has necessitated the development of antibiotic alternatives. However, relatively little consideration has been given to the development of resistance to these alternatives.Whether we come up with antibiotic alternatives that are bacteriocidal or inhibitory, bacteria will continue to adapt and evolve.Some antibiotic alternatives support the development of antibiotic resistance necessitating caution.There are opportunities to optimize antibiotic alternative effectiveness as well as to minimize the development of resistance mechanisms.With the growing concern of antibiotic resistance , there hCampylobacter jejuni  system as a pseudoimmune system, or the abortive infection system to kill infected cells before the phage can spread , Zinc (Zn2+), and Silver (Ag+), and nonmetal elements, such as Iodine (I2), have been used in animal production for their broad-spectrum antibacterial activity and low generation of resistance  with a wolf in sheep’s clothing .Pig image: many antimicrobial alternatives target the early post-weaning piglet.Polyphenols: polyphenolic compounds found in plant products, such as these pea seed coats, have strong antibacterial activity.


Curcuma longa L, which possesses a wide range of pharmacological effects. However, few studies have collected scientific evidence on its dual effect on angiogenesis. The present review gathered the fragmented information available in the literature to discuss the dual effect and possible mechanisms of curcumin on angiogenesis. Available information concerning the effect of curcumin on angiogenesis is compiled from scientific databases, including PubMed and Web of Science using the key term (curcumin and angiogenesis). The results were reviewed to identify relevant articles. Related literature demonstrated that curcumin has antiangiogenesis effect via regulating multiple factors, including proangiogenesis factor VEGF, MMPs, and FGF, both in vivo and in vitro, and could promote angiogenesis under certain circumstances via these factors. This paper provided a short review on bidirectional action of curcumin, which should be useful for further study and application of this compound that require further studies.Curcumin is a compound extracted from the Curcuma longa L with the symmetric chemical structure [Curcumin, one of the most promising natural compounds, is the major polyphenol compound found intructure , 2. Sinctructure , Lampe etructure , and idetructure ; extensitructure –11. Prectructure . Cancer Angiogenesis is the process of new vessel formation and hallmark of tumor progression , which iThis article is aimed at reviewing the bidirectional action of curcumin and curcuminoids as well as synthetic curcumin analog on angiogenesis based on current research findings, focusing on regulation of curcumin on proangiogenesis factors in antiangiogenesis and proangiogenesis process; some of the mechanisms were summarized and discussed. Furthermore, this article provided some information and insights which could be interesting to researchers in related areas. Curcuma longa L has been used as a commonly used traditional Chinese medicine for thousands of years with the effect of activating qi flowing, removing blood stasis, benefiting menstruation, and relieving pain. In India, turmeric is commonly used in the Indian subcontinent as a spice, concurrently used for health care including various respiratory diseases [Curcumin  is a memdiseases , 23. Besdiseases .α,β-unsaturated β-diketone [β-diketone moiety, curcumin exists in keto-enol tautomers, and this tautomerism favors interaction and binding with a wide range of enzymes [κB) differ between curcuminoids, suggesting that the methoxy groups on the phenyl rings in curcumin are important to have health effects [Chemically, curcumin is a bis-diketone   can be inhibited by curcumin both in vitro and in vivo, which has a significant role in angiogenesis [ in vitro and vivo; results showed decreasing of microvessel density, which is regulated by targeting the nuclear factor-κB pathway [κB in corneal cells induced by lipopolysaccharide [κB activation in mediating angiogenesis and curcumin could downregulate NF-κB to inhibit angiogenesis.NF-tivation . Activatogenesis , 79; the pathway . Curcumiccharide . These sDespite the antiangiogenesis of curcumin has been discussed a lot; it is found that curcumin possess a proangiogenesis effect. Study found out that curcumin pretreatment augmented adipose derived stem cells production of VEGF, which contributed to neovessels formation and improving cells survival . A previ in vivo effects of curcumin on wound healing in rats and guinea pigs has been studied showing that extensive neovascularization and molecular biology analysis also showed an increase in the mRNA transcripts of TGF-beta1 in curcumin-treated wounds [Meanwhile, neovascularization represents an important part in wound healing; angiogenesis could affect the whole process of wound healing from the very beginning after skin injury until the end of the wound remodeling ; curcumid wounds . These r Curcuma longa also showed significant proangiogenesis activity [Expect for curcumin itself, ethosomal curcumin also showed the effect of promoting neovascularization in the second degree burns in rat  and curcactivity . All tog in vitro study showed that curcumin reveals opposite angiogenesis effects on human umbilical vein endothelial cells and chicken chorioallantoic membrane as a function of dose [As discussed before, it is found that curcumin shows a dual effect on angiogenesis ; studies of dose . Besides of dose . Proangi of dose . It was  of dose , while i of dose , indicatThe inhibition of angiogenesis differentiation by curcumin depends on the serum concentration present in the incubation medium. The effective concentration required to inhibit angiogenesis in the presence of 10% serum is much higher than that required to do so in the presence of 2% serum .As reviewed in this article, it can be seen that curcumin has dual effects on angiogenesis; curcumin can not only inhibit angiogenesis in tumor and other pathological conditions, but also promote angiogenesis in fresh wound and diabetic rats, which has broadened the clinical use, pharmaceutical significance, and therapeutic applicability of curcumin, providing a new research direction on curcumin.Even though the dual effect of curcumin has been found during years of hard work, the molecular mechanism of its dual effects has been studied in some details but the underlying mechanism of this association between angiogenesis and the dual effect of curcumin is still not crystal clear. Some researches proposed the idea that the dual effect of curcumin is dose-based or depending on the microenvironment, there is other hypothesis that chemistry structure of curcumin may contribute to it, but no solid conclusion has been drawn; further researches based on chemical genomics approach and other methods are still needed.κB pathway. While curcumin shows a two-way prooxidant and antioxidant effects regulated by concentration [κB pathway plays an important role as well, indicating that the underlying mechanisms of bidirectional actions of curcumin on angiogenesis are more complicated.Such bidirectional actions of curcumin are not uncommon. Curcumin has radioprotective effects on normal tissues and radiosensitization of tumor cells –101, whintration –104, theThere is study which showed microRNAs as a part of cellular communication . Researc in vitro and in vivo, and curcumin has been used in traditional medicine for many years with accurate effects and has tremendous potential in treating difficult disease; it still needs deeper researches, both evidence-based preclinical and clinical researches for evaluating its pharmaceutical potentialities and better understanding of its pharmacological mechanisms, and puts studies of curcumin translate from lab to clinical. However there are limitations to this research; there are still many deficiencies in the thesis. It is our hope that more people see the value in this natural compound and provide evidence needed to help prove its mechanisms.Recent researches focus mostly on the unilateral role of curcumin on angiogenesis; studies focused on antiangiogenesis effect of curcumin showed that curcumin has great potential in anticancer treatment, as a natural product with great safety and low toxicity; more preclinical and clinical studies should be conducted to dig the anticancer effect of curcumin through inhibiting angiogenesis and promote new drug research and development. Novel delivery systems of curcumin have been investigated extensively to improve the bioavailability


The scope of targets investigated in pharmaceutical research is continuously moving into uncharted territory. Consequently, finding suitable chemical matter with current compound collections is proving increasingly difficult. Encoded library technologies enable the rapid exploration of large chemical space for the identification of ligands for such targets. These binders facilitate drug discovery projects both as tools for target validation, structural elucidation and assay development as well as starting points for medicinal chemistry. Novartis internalized two complementing encoded library platforms to accelerate the initiation of its drug discovery programs. For the identification of low-molecular weight ligands, we apply DNA-encoded libraries. In addition, encoded peptide libraries are employed to identify cyclic peptides. This review discusses how we apply these two platforms in our research and why we consider it beneficial to run both pipelines in-house. This lowers the chances to identify hits with high affinity and selectivity or for demanding targets. LMW agents originating from traditional compound libraries may in principle be bioavailable, allow for cell penetration and are suited to bind in defined pockets of proteins such as enzymes. Yet they frequently perform worse against protein-protein interactions (PPIs). PPIs tend to have rather large and extended interaction surfaces. On the other side of the spectrum, biologics are better suited for such applications. They are able to address extended PPIs on large surfaces and are usually both very selective and potent. However, their large molecular weight, amongst others, largely excludes addressing intracellular targets in a therapeutic manner as well as oral administration. Advances in disease characterization and target identification ,2 consta9 and more possible chemical entities. This size thus is 1’000-fold larger than a usual compound collection of pharmaceutical companies and significantly increases the chances of finding new chemical starting points. As a second pillar, encoded peptide libraries as utilized in the Novartis-internal Peptide Discovery Platform (PDP) can fill the gap between LMW and biologics. Peptides with a typical length of 5–20 amino acids have an intermediate size and are able to interact with target proteins in many ways, like covering large surfaces or small binding pockets. Depending on the set-up, peptidic libraries can encode up to 1012 different sequences, allowing for identification of high affinity binders against almost any target [Encoded libraries can help to overcome these shortcomings. Compounds originating from DNA-encoded libraries (DEL) resemble LMW molecules, but offer the opportunity to screen up to 10y target .First theoretically proposed by Brenner and Lerner in 1992 , the selThere are hundreds of different proprietary DELs used by the individual labs and institutions. Despite their distinct designs, most of them employ the same common “split-and-pool” approach for the generation of chemical diversity on a massive scale ,12. ThisBy combining innovative library design based on the constant development of new chemistries ,16,17,18The standard DEL selection procedure has been reviewed numerous times in the past ,20,21, tOver the past decade, various developments in the DEL field have caused this technology to become a well-established tool that requires little project-specific assay development. Therefore, DEL screens provide a time- and cost-effective entry point into new projects, supporting aspects from target validation to hypothesis testing and hit discovery. Once nanomolar quantities of the purified protein of interest (POI) are available, it is straightforward to assess the general ligandability of the target. As the DEL assay format allows simultaneous screening through parallel automation with multi-channel pipettes or alternative liquid handlers ,24, vari7 [Peptidic ligands offer an attractive alternative to LMW compounds. A comprehensive overview of the various technologies to identify peptides can be found in recent reviews ,26. All 7 . 9 sequences [Escherichia coli is generally limited to the incorporation of the 20 natural amino acids and cyclization is mainly possible through disulfide bridges only. However, stably cyclized peptides are often favorable to e.g., prevent proteolysis. In addition, ring closure reduces conformational freedom and thus results in peptides with higher affinity and selectivity compared to their linear counterparts [To increase the variety of peptides, molecular biology based selection approaches were introduced, linking the phenotypic (peptide sequence) to the genetic (coding) information provided by the nucleic acid sequence. The scope of phage display techniques , best knequences ,30. Phagterparts . These lterparts ,33,34, eterparts . 9 to 1013 possible sequences, as libraries are translated in vitro, making the diversity limiting step of cell transformation unnecessary. Since the encompassment by phage particles is lacking, the linkage between expressed peptide and nucleic acid must happen during the translation process in order to ensure an unambiguous linkage. One of the most widespread approaches to fulfill this requirement is mRNA display, in which the antibiotic puromycin is attached to the mRNA strand and leads to a covalent link between the mRNA  system such as the “Protein synthesis Using Recombinant Elements” (PURE) was introduced by the Ueda lab from the University of Tokyo [To cover a maximal chemical space of displayed peptides, the so-called “reprogramming” of the genetic code, i.e., enabling the incorporation of non-natural amino acids, was the last key advancement towards the versatile Peptide Discovery Platform as it is used now at Novartis ,40. Tradof Tokyo . This coof Tokyo . These sof Tokyo ,46. By jof Tokyo ,47. ThisAs described above for DEL screens, consecutive selection rounds based on the affinity of the library members to the POI are also applied with the peptide discovery technology C. In conBoth DEL and PDP screens offer various benefits over conventional approaches like HTS for LMW compounds or phage display for peptide discovery, thereby enabling rapid access to both tool compounds as well as suitable starting points for drug discovery projects. First of all, the sizes of the respective input libraries are larger, allowing the chemical space to be more deeply sampled in just one screen. Additionally, in comparison to other display systems (like phage or yeast), less library focusing is inherent to these cell-free techniques, i.e., less reduction of the starting pool’s diversity due to different growth rates of the employed cellular systems. Due to the activity-agnostic characteristic of the ELT technology, chemical matter for diverse MoAs can be identified with only limited project-specific assay development. As the screens are neither focusing on the target’s known biochemical activity nor its interaction partners, their output can also include “silent binders”. Such binders might be of great interest as tool compounds or be employed in projects focusing on applications other than classical drug discovery, like protein degradation or peptide-drug conjugates ,49.Because of the combinatorial nature of the employed libraries, DEL hits usually can be identified on different “levels”: Often, not only individual ligands are identified, but rather larger families can be detected, in which related building blocks or combinations thereof are enriched . This feThe fact that only limited assay development and rather low amounts of reagents (regarding both targets and libraries) are required often makes ELT screens the methods of choice for the first round of identifying binders in the pharmaceutical industry, particularly when tackling novel targets. Another key benefit of ELT is the ability to perform several selections in parallel: by simultaneously screening against multiple targets, different protein constructs and/or various selection conditions, different MoAs can be probed in a cost- and time-effective way (and not in a linear flowchart as most often done in standard HTS projects). As any selection system, ELTs also hold certain challenges and limitations. Direct functional read-outs can only be implemented after laborious technical development, mainly for two reasons: On the one hand, in both selection systems the respective targets are present in great excess over the individual ligands, making it challenging for enzymatic assays to give rise to tangible results. On the other hand, both in DEL and PDP screens enrichment is driven by the binding affinities and kinetics of their library members to the respective target and not based on functional effects. Thus, identified hits generally need to be thoroughly validated in subsequent assays, which requires them to be resynthesized in an untagged format in sufficient quantities. This hit re-synthesis is also important to unambiguously identify the interacting chemical matter, as despite thorough synthesis validations, the occurrence of side reactions in the library preparation cannot be fully excluded. Several methods were developed to enable the facile validation of large numbers of hits on DNA ,51. Nonede facto inverting the synthesis procedure and attaching the DNA to the other end of the tagged ligand. However, very deep or fully enclosed pockets may remain out of reach. An overview of all mentioned advantages and challenges for encoded library technologies in comparison to conventional HTS is provided in Generally, the barcodes of the library members impose an additional multitude of limitations on ELTs. Most obvious is the scope of chemical reactions compatible with DNA: even though multiple new chemical reactions are reported regularly, some conditions remain challenging, e.g., those containing strong acids, requiring extensive UV-light usage or demanding water-free conditions . For PD9 individual compounds. Once available, these libraries can be used for hundreds to thousands of selection campaigns. A DEL screen typically consists of two to three rounds of panning which can be completed in a few days. Together with sample preparation for sequencing, an entire DEL screen can be conducted in about one week. In contrast, a PDP screen typically takes several weeks as both an individual selection round is more labor intensive and more rounds need to be conducted (up to 10). The reason for this greater cycle number is based on a number of key differences between the two technologies. Firstly, for PDP the theoretical library size based on the degenerate code is frequently greater than the main limiting factor in the entire mix: the number of active ribosomes, which is estimated to be around 1012–1013. Consequently, each peptide sequence is statistically present as a single instance in the input library. Thus, binders can only be identified after several rounds of selection cycles. Secondly, the DNA sequences are amplified and the peptide library is regenerated at every selection cycle. This allows positive binding sequences to be enriched over consecutive cycles while keeping the total amount of library constant. This is a clear advantage over DEL where ligand enrichment takes place at the expense of total library material. As a result, PDP selections can readily be run until individual peptide sequences dominate the pool. Additionally, this enables step-wise altering the selection conditions between the rounds or revisiting earlier rounds if required. In analogy to DEL, the procedure itself can be streamlined and at least partly automated on pipetting systems which permits the screening of several targets and/or selections conditions in parallel [When comparing the two technologies with each other they both offer their own advantages. In the case of PDP, library generation by IVT can be accomplished in less than one hour once all reagents are available and hence can be performed at each round of selection. In contrast, DEL synthesis needs significant effort to generate the desired size of 10parallel . This en12. In spite of their potency, peptide hits are rarely applied as immediate starting points for intra-cellular targets due to their low cell-penetration although some possibilities to tweak their properties in the desired direction exist [As will be discussed in detail in the following section a major difference is in the application scope of the identified ligands. DEL screening hits tend to have affinities in the low µM range. However, they readily serve as suitable starting points for further medicinal chemistry efforts. In contrast, the peptidic hits identified through PDP usually exhibit binding in the nM range. This increased affinity can be rationalized due the larger interaction surfaces of those ligands as well as the greater number of library members of about 10on exist . Alternaon exist ,58. OnlyMajor pharmaceutical companies have multiple hit finding technologies at their disposition when initiating drug discovery programs. Several factors determine which of these technologies are applied in which order to a given target. A more complete overview of all considerations will take place at a later stage of this review. When deciding between the two encoded library platforms discussed in this review, one important scientific criterion to consider is the purpose of the ligand being sought. Most DELs are designed to contain a large fraction of LMW molecules within a favorable property space for initiating medicinal chemistry campaigns towards orally bioavailable drugs. In a first pass, ligands identified from DEL screens are thus geared towards the same target application scope as hits from a similarly tailored compound collection used for HTS campaigns. Consequently, screening of DELs is the preferred option over peptide libraries if a molecule with more drug-like properties is the immediate aim of the experiment.Two examples recently reported by GSK for receptor-interacting serine/threonine-protein kinase (RIPK1)  and soluThe scope of applications of DEL hits is not limited to serving as LMW starting points. Due to the nature of combinatorial chemistry, not every library member fully adheres to the desired property space aimed for when designing the DEL. Such compounds may still provide important information on challenging targets where other hit finding efforts do not lead to suitable ligands. In a recent example, a selective ligand termed BAY-850 for the epigenetic regulator protein ATPase family AAA-domain containing protein 2 (ATAD2) was identified in screens against its bromo-domain . In spit50 of 2 µM. A co-crystal structure revealed that the ligand bound in a β-turn conformation, causing the two ends of the peptide to be in proximity of each other. Macrocyclization of this compound enabled the design of a highly selective inhibitor with an IC50 of <3 nM. DEL technology is also compatible with directly synthesizing macrocyclic libraries either by combinatorial assembly of the ring during its synthesis [12 members composed of 4–20 amino acids. Such design in essence forms a bridge between typical DELs and PDP libraries [As stated above, some of the key advantages of the DEL technology include the comparably rapid access to very large compound collections and the possibility to pool multiple libraries in one screen. This provides the opportunity to build DELs addressing certain areas of chemical space aiming e.g., at identifying tool compounds rather than drug-like hits. One common addition to any DEL portfolio is frequently referred to as “capped dipeptide” or “tripeptide library”. This synthesis procedure connects three building blocks by two amide bonds in a linear fashion. The advantages of such a set-up include the vast availability of building blocks as well as the efficiency of the bond forming reaction. Such DELs reliably deliver validated hits and can correspondingly serve the purpose of assessing general ligandability of a target. An example on how to progress such compounds was recently reported for the induced myeloid leukemia cell differentiation protein . A DEL synthesis  or the dynthesis . A notewibraries .Employing DEL screening to assess ligandability of a given target is further facilitated by the possibility of combining multiple libraries in one screen. Pooling DELs based on different synthetic strategies in one experiment allows efficient probing of the target for accessible binding pockets. Two recent reports on protease-activated receptor (PAR2) exemplify this well. Screening of DELs against a thermo-stabilized mutant of the receptor identified two ligand families binding to distinct sites on the target . The firIn contrast to DEL, the vast majority of hits identified using PDP are neither cell permeable nor orally bioavailable without substantial optimization. However, the platform very reliably delivers potent ligands against challenging targets or MoAs for which other hit finding methods are traditionally less successful, such as membrane proteins and PPIs. Thus, it is not surprising that these peptides routinely find applications as tools to enable drug discovery projects. Due to their peptidic nature, protein-protein-interactions represent an obvious MoA to be targeted by this class of ligands. A recent example in literature describes the identification of potent macrocyclic inhibitors of the interaction between the Zaire Ebola virus protein 24 and karyopherin alpha 5 . AlthougPyrococcus furiosus multidrug and toxic compound extrusion transporter identified four inhibitory macrocycles. These ligands facilitated structural studies of this protein by co-crystallization and were shown to bind to different regions of the target, locking it in an outward-facing conformation [p-glycoprotein homolog from Cyanidioschyzon merolae co-crystallized with its target in an inward open conformation. Comparison with the apo-structure gave further insights into the mechanism of this transporter [An additional tool application of PDP-derived ligands is enabling structural determination of hard to crystallize proteins as well as giving rise to structures in different conformations. Noteworthy examples are two membrane transporter proteins: Firstly, screening of peptide libraries against ormation ,71. Seconsporter . Due to their high affinity and good selectivity, PDP ligands can also serve as probes to study the function of their target. This was recently exemplified by a screen of the catalytic domain of Lysine-specific demethylase 4A, which identified ligands with novel binding modes . These i9 bicyclic peptides [Despite the challenges with respect to oral bioavailability and cell penetration, ligands identified through PDP and related technologies ,74 may apeptides ,76. In apeptides ,78. Bothpeptides .in vivo and results in silencing of the targeted genes [Another means of expanding the therapeutic scope of peptidic ligands can be achieved by the conjugation of functional payloads, generating peptide-drug conjugates (PDCs) . In comped genes .ELTs are most successful when applied to soluble, well-behaved target proteins. This is readily rationalized as in the most commonly applied screening methods ligand enrichment relies on the stable biophysical interaction with the target and is driven by the target concentration. Consequently, poorly structured proteins or targets adopting multiple conformations result in lower enrichment. This may also contribute to the observation that a larger number of DEL literature reports describe the identification of enzyme inhibitors binding to well defined pockets . NonetheTo date, larger protein complexes as well as transcription factors are target classes, which are still hard to address with DEL and PDP. For the latter of these targets, the presence of DNA-binding domains and largely unstructured domains represent the major obstacles. For complexes, a multitude of challenges ranging from protein production, target complex stability and immobilized amounts as well as more opportunities for unspecific binding due to increased target size are conceivably responsible for the lack of success. Some of the abovementioned limitations may be overcome when using covalently binding ligands. In this case, enrichment is not primarily governed by reversible binding constants, making the identification of less frequently occurring binding events possible ,98. Bothin silico methodologies and medicinal chemistry into one project framework. This enables multidisciplinary project teams to effectively react on findings that cross-fertilize all other involved disciplines. Often enough, the successful drug candidates are strongly influenced by the various efforts. As a consequence, the chemical matter is not a product of one certain technology, but the joint invention of the whole multidisciplinary team. With such a set-up, any potential competition between either the approaches, involved partners or sub-teams is transformed into collaboration.Novartis strategically conducts small molecule lead discovery projects in a holistic manner termed “integrated lead finding” . This coAs drug discovery is an innovative and evolving field, pharmaceutical companies constantly need to reflect on their lead discovery technology arsenal to remain effective and successful. Novel drug finding technologies are often invented and first applied in academic or biotech settings. This also holds true for DEL and PDP screening where multiple biotech and academic groups have specialized their efforts in. The PDP as described above was first commercialized by the Japanese biotech company “PeptiDream” and a dedicated DEL platform was developed by the Danish biotech company “Nuevolution”. To assess the value of both lead finding technologies, Novartis successfully collaborated with these biotech partners in a step-wise approach. Initial feasibility studies revealed the great potential and complementary strengths of both technologies. This led to project-based ligand-finding collaborations. Both milestone-driven partnerships delivered tangible chemical matter in a number of projects and the technologies were thus internalized to facilitate a broader and fully enabled deployment within Novartis’ project portfolio.Integrated lead finding requires effective communication, hand-in-hand cooperation as well as full and timely data sharing between all involved sub-teams. This can be more challenging in external collaborations, particularly when the complexity of the projects requires iterative and multidisciplinary lead finding efforts. Therefore, our approach was to internalize both technologies in-house to effectively enable and support the Novartis project portfolio. Besides that, internalization allowed Novartis to deploy the technologies to any internal project without potential limitations of the technology partner company due to other competing agreements. Another important reason for the chosen business model was the freedom to internally build up an individualized and proprietary library portfolio that best suits Novartis’ needs. Lastly, the technologies required specific adaptations and innovations to fully fit to Novartis’ scientific requirements.Through the internalization, these encoded technologies are fully embedded in Novartis’ lead finding efforts and are operated by teams with multidisciplinary backgrounds. The DEL and PDP platforms have strong synergies, both benefiting from a close collaboration and therefore optimally intertwine. Many of the involved technology aspects are synergistic and require similar expertise and set-up. The two screening platforms were grown within the screening sciences discipline and possess a shared set-up between the chemical biology and discovery chemistry groups—this interdisciplinary strategy is fundamental for their impact and ensures best leverage of the available in-house drug discovery and medicinal chemistry knowledge.As discussed earlier, both technologies, DEL and PDP, offer various opportunities to augment more classical drug discovery approaches and to deliver complementing, synergizing chemical matter. For Novartis, DELs nicely expands other available low molecular weight screening decks, like those for fragment based and medium/high-throughput screening. Besides their high value for hit finding, the classical screening decks are also a fundamental source of proprietary building blocks to assemble new and innovative DELs with unique chemical matter. On the other side, resynthesized DEL compounds are continuously merged into the classical screening decks and thus made available for any future conventional hit finding effort.In many of our encoded library projects, the screening results enable the design of novel or additional lead finding efforts. PDP has a strong track record in efficiently identifying ligands with high affinity and target selectivity. These represent valuable starting points for extracellular targets or can serve as high impact tools for structural efforts like the exploration of binding pockets or pharmacophore mapping. Another important application of PDP-derived ligands is their use as tools to identify small molecule chemical matter in competition screens. This was a significant bottleneck before encoded library technologies became available. Designing and developing assays to identify chemical matter for early projects was a sometimes both time-consuming and resource-intense effort, particularly in absence of known target ligands. Identifying low molecular target binders at Novartis was restricted to relatively resource intense biophysical methodologies like NMR  or MS-baToday, we apply encoded library screening to novel targets in a way that is agnostic to already available tool compounds. Particularly PDP rapidly identifies suitable high affinity binders, possibly to multiple pockets on the individual targets. In few iterations, we can investigate key motifs responsible for affinity, e.g., by amino acid scanning. In most cases, it is straightforward to attach a fluorophore on the ligand at positions not involved in target binding—often the linkage vector to the encoding tags is readily available and suitable for tethering. Using such ligands as tools for competition assays is helpful to decipher the binding mode for chemical matter. In principle, DEL can also identify such tools, however as previously stated PDP has a higher likelihood to identify binders with the required affinity.Particularly for early stage drug discovery projects both technologies help addressing key scientific questions associated with the biological context, the disease relevance or which sites on the target are associated with the desired functional effect. As an example, a recent DEL campaign against a target hypothesized to be involved in cardiomyopathy revealed the role of individual domains in contraction and relaxation of diseased heart muscle. Initial parallel DEL screening of multiple protein domains and their combinations indicated the relevance of specific combinations. This information was vital to confirm the target hypothesis and to design a suitable lead finding strategy and project flow chart. It also permitted the identification of ligands that showed functional activity and proof of concept in a bio-artificial muscle contractility assay. Similarly, several PDP campaigns strongly impact projects through the identification of unprecedented high affinity ligands for novel and often induced pockets. The compounds enable co-crystal structures and pharmacophore mapping for new MoAs or highlight the therapeutic potential of novel pockets due the observed functionality or selectivity. Acquiring these detailed insights can be vital to design the most appropriate project flowchart.As described above, ELTs are agnostic to functional target activity and can thus be applied to a variety of possible drug discovery projects. Suitable encoded library screening projects are picked based on a science- and priority-driven assessment. Chemistry and biology ELT experts discuss and align with key project scientists how the strengths of both technologies can impact the individual project in the most efficient manner. This discussion is influenced both by other foreseen lead finding efforts and the overall goal of the project. Balancing this with resource distribution and Novartis’ priorities ensures that DEL and PDP screening is applied with highest impact on the portfolio in the framework of integrated lead finding. Still, for several projects both platforms are applied if the anticipated impact, the project-priority as well as scientific questions benefit from such an approach.Project campaigns are tactically binned to minimize the timeframe from tool availability to the generation of ligand hit lists for the projects. This empowers the ELT team to identify possible ligands within 2–4 months from the beginning of wet-lab activities, including technology specific assay development, preparation and screening, sequencing, data parsing and eventually hit analysis. In the past years, through continuous technical and strategic optimizations, we have achieved a strong project impact track record: PDP delivered validated high affinity ligands in >85% of the performed projects while DEL screens identified validated chemical matter for >66% of the selections. These projects included many challenging targets with unprecedented chemical matter or those that were a challenge for encoded technologies .Other pharmaceutical companies successfully apply similar technologies with a different model compared to Novartis, e.g., by internalizing full biotech companies specialized on technologies like DEL, or by fully outsourcing certain project aspects  to biotech collaboration partners. Besides Novartis, several large pharmaceutical companies in-licensed encoded library technologies or have built-up their own capacities in the field. To our knowledge, however, Novartis is uniquely applying both screening technologies in a synergistic and combined set-up as an integrated approach while other players in the field only apply either DEL or PDP. In our eyes, fully embedding both methodologies into Novartis’ drug discovery teams with enabled expert scientists shows great impact and ensures effective and seamless collaborations as described above.Over the last decade, ELTs emerged as powerful method to rapidly access large chemical space using comparably simple affinity screening. As stated earlier, one of the major limitations of ELTs is that the most frequently applied selection methodologies primarily enrich ligands based on binding to the target protein. While variations of the conditions allow the detection of differences in affinity and selectivity  or may hint at possible binding sites , a direct functional read-out remains challenging. Still, a few examples on how to tackle this limitation were described recently: For peptide libraries, SICLOPPS enables functional in-cell selections of peptides by coupling the desired activity to the survival of the cell . For smaAnother technological aspect of these platforms to likely see major advances in the next years is data analysis. The read-outs of DEL and PDP screening—typically performed by NGS—generate large amounts of data. The reported analysis methods mainly focus on the similarity of enriched building blocks or their combinations as well as the performance of the enriched ligands under the different screening conditions . HoweverThe application scope of ligands originating from ELT will most likely expand in the near future as well. In the recent years, a number of modalities gained attention in which molecules exhibiting different functions are chemically connected. This includes the abovementioned PDCs, but also bifunctional degraders . LigandsThese and other future optimizations will further increase the impact of encoded library technologies on drug discovery. The complementing in-house availability of the two platforms discussed in this review allows Novartis to apply them synergistically to our projects and to identify ligands against almost any target in a way complementary to more conventional HTS or fragment based methods. Due to the large chemical space covered, these platforms enable us to rapidly respond to both the evolving targets space and novel modalities in an efficient and thorough way. Thus, the full embedding of both platforms into our integrated lead finding concept allows us to broadly deploy them to the Novartis project portfolio and to maximize their impact on future drug discovery.