How is who: evidence as clues for action in participatory sustainability science and public health research.

Participatory and collaborative approaches in sustainability science and public health research contribute to co-producing evidence that can support interventions by involving diverse societal actors that range from individual citizens to entire communities. However, existing philosophical accounts of evidence are not adequate to deal with the kind of evidence generated and used in such approaches. In this paper, we present an account of evidence as clues for action through participatory and collaborative research inspired by philosopher Susan Haack's theory of evidence. Differently from most accounts of evidence for use in policies and interventions, our account combines action-oriented (the how) and actors-oriented (the who) considerations. We build on Haack's theory and on the analysis of examples of participatory and collaborative research in sustainability science and public health research to flesh out six procedural criteria for the generation and mobilization of evidence in and from participatory research. Action-oriented criteria invite to look at evidence from a (a) foundherentist, (b) gradational and (c) quasi-holistic perspective. Actors-oriented criteria point out that evidence generation and utilization are (d) social, (e) personal, and (f) embedded. We suggest that these criteria may reinforce participatory and collaborative approaches to evidence co-production when addressing complex problems in sustainability science and public health allowing for the generation of a kind of practical objectivity.

Complexity in Epidemiology and Public Health. Addressing Complex Health Problems Through a Mix of Epidemiologic Methods and Data.

Public health and the underlying disease processes are complex, often involving the interaction of biologic, social, psychologic, economic, and other processes that may be nonlinear and adaptive and have other features of complex systems. There is therefore a need to push the boundaries of public health beyond single-factor data analysis and expand the capacity of research methodology to tackle real-world complexities. This article sets out a way to operationalize complex systems thinking in public health, with a particular focus on how epidemiologic methods and data can contribute towards this end. Our proposed framework comprises three core dimensions-patterns, mechanisms, and dynamics-along which complex systems may be conceptualized. These dimensions cover seven key features of complex systems-emergence, interactions, nonlinearity, interference, feedback loops, adaptation, and evolution. We relate this framework to examples of methods and data traditionally used in epidemiology. We conclude that systematic production of knowledge on complex health issues may benefit from: formulation of research questions and programs in terms of the core dimensions we identify, as a comprehensive way to capture crucial features of complex systems; integration of traditional epidemiologic methods with systems methodology such as computational simulation modeling; interdisciplinary work; and continued investment in a wide range of data types. We believe that the proposed framework can support the systematic production of knowledge on complex health problems, with the use of epidemiology and other disciplines. This will help us understand emergent health phenomena, identify vulnerable population groups, and detect leverage points for promoting public health.

Opitz syndrome: improving clinical interpretation of intronic variants in MID1 gene.

BACKGROUND: Loss-of-function variants in MID1 are the most common cause of Opitz G/BBB syndrome (OS). The interpretation of intronic variants affecting the splicing is a rising issue in OS. METHODS: Exon sequencing of a 2-year-old boy with OS showed that he was a carrier of the de novo c.1286-10G>T variant in MID1. In silico predictions and minigene assays explored the effect of the variant on splicing. The minigene approach was also applied to two previously identified MID1 c.864+1G>T and c.1285+1G>T variants. RESULTS: Minigene assay demonstrated that the c.1286-10G>T variant generated the inclusion of eight nucleotides that predicted generation of a frameshift. The c.864+1G>T and c.1285+1G>T variants resulted in an in-frame deletion predicted to generate a shorter MID1 protein. In hemizygous males, this allowed reclassification of all the identified variants from "of unknown significance" to "likely pathogenic." CONCLUSIONS: Minigene assay supports functional effects from MID1 intronic variants. This paves the way to the introduction of similar second-tier investigations in the molecular diagnostics workflow of OS. IMPACT: Causative intronic variants in MID1 are rarely investigated in Opitz syndrome. MID1 is not expressed in blood and mRNA studies are hardly accessible in routine diagnostics. Minigene assay is an alternative for assessing the effect of intronic variants on splicing. This is the first study characterizing the molecular consequences of three MID1 variants for diagnostic purposes and demonstrating the efficacy of minigene assays in supporting their clinical interpretation. Review of the criteria according to the American College of Medical Genetics reassessed all variants as likely pathogenic.

Loss-of-function variants in exon 4 of TAB2 cause a recognizable multisystem disorder with cardiovascular, facial, cutaneous, and musculoskeletal involvement.

PURPOSE: This study aimed to describe a multisystemic disorder featuring cardiovascular, facial, musculoskeletal, and cutaneous anomalies caused by heterozygous loss-of-function variants in TAB2. METHODS: Affected individuals were analyzed by next-generation technologies and genomic array. The presumed loss-of-function effect of identified variants was assessed by luciferase assay in cells transiently expressing TAB2 deleterious alleles. In available patients' fibroblasts, variant pathogenicity was further explored by immunoblot and osteoblast differentiation assays. The transcriptomic profile of fibroblasts was investigated by RNA sequencing. RESULTS: A total of 11 individuals from 8 families were heterozygotes for a novel TAB2 variant. In total, 7 variants were predicted to be null alleles and 1 was a missense change. An additional subject was heterozygous for a 52 kb microdeletion involving TAB2 exons 1 to 3. Luciferase assay indicated a decreased transcriptional activation mediated by NF-κB signaling for all point variants. Immunoblot analysis showed a reduction of TAK1 phosphorylation while osteoblast differentiation was impaired. Transcriptomic analysis identified deregulation of multiple pleiotropic pathways, such as TGFß-, Ras-MAPK-, and Wnt-signaling networks. CONCLUSION: Our data defined a novel disorder associated with loss-of-function or, more rarely, hypomorphic alleles in a restricted linker region of TAB2. The pleiotropic manifestations in this disorder partly recapitulate the 6q25.1 (TAB2) microdeletion syndrome and deserve the definition of cardio-facial-cutaneous-articular syndrome.

Sprouts of Moringa oleifera Lam.: Germination, Polyphenol Content and Antioxidant Activity.

(1) Background: In recent years, the consumption of sprouts, thanks to their high nutritional value, and the presence of bioactive compounds with antioxidant, antiviral and antibacterial properties, is becoming an increasingly widespread habit. Moringa oleifera Lam. (Moringa) seems to be an inexhaustible resource considering that many parts may be used as food or in traditional medicine; on the other hand, Moringa sprouts still lack a proper characterization needing further insights to envisage novel uses and applications. (2) Methods: In this study, a rapid and easy protocol to induce the in vivo and in vitro germination of Moringa seeds has been set up to obtain sprouts and cotyledons to be evaluated for their chemical composition. Moreover, the effects of sprouts developmental stage, type of sowing substrate, and gibberellic acid use on the chemical characteristics of extracts have been evaluated. (3) Results: Moringa seeds have a high germinability, both in in vivo and in vitro conditions. In addition, the extracts obtained have different total phenolic content and antioxidant activity. (4) Conclusions: This research provides a first-line evidence to evaluate Moringa sprouts as future novel functional food or as a valuable source of bioactive compounds.

Surface-clearance levels for the normal reuse of objects for 413 radionuclides.

This paper provides surface-clearance levels for 413 radionuclides for the normal reuse of objects leaving the controlled area of a nuclear facility. Four sets of surface-specific clearance levels are derived based on choices of dose coefficients (DCs) in probabilistic dose assessments using the SUrface DOse QUantification model (SUDOQU; van Dillen and van Dijk 2018J. Radiol. Prot.381147-203). The general dose criteria for clearance, extended with a suitable dose constraint for the concept of the representative person, are applied in the derivations. These clearance levels apply to the total of fixed and removable radioactive contamination on the surface of objects. The results are discussed in the context of conservatism of the exposure scenarios (parameters, DCs), and guidance is given on the selection and application of the values.

COVID-19 heralds a new epistemology of science for the public good.

COVID-19 has revealed that science needs to learn how to better deal with the irreducible uncertainty that comes with global systemic risks as well as with the social responsibility of science towards the public good. Further developing the epistemological principles of new theories and experimental practices, alternative investigative pathways and communication, and diverse voices can be an important contribution of history and philosophy of science and of science studies to ongoing transformations of the scientific enterprise.

D-Amino acids in mammalian endocrine tissues.

D-Aspartate, D-serine and D-alanine are a regular occurrence in mammalian endocrine tissues, though in amounts varying with the type of gland. The pituitary gland, pineal gland, thyroid, adrenal glands and testis contain relatively large amounts of D-aspartate in all species examined. D-alanine is relatively abundant in the pituitary gland and pancreas. High levels of D-serine characterize the hypothalamus. D-leucine, D-proline and D-glutamate are generally low. The current knowledge of physiological roles of D-amino acids in endocrine tissues is far from exhaustive, yet the topic is attracting increasing interest because of its potential in pharmacological application. D-aspartate is known to act at all levels of the hypothalamus-pituitary-testis axis, playing a key role in reproductive biology in several vertebrate classes. An involvement of D-amino acids in the endocrine function of the pancreas is emerging. D-Aspartate has been immunolocalized in insulin-containing secretory granules in INS-1 E clonal ß cells and is co-secreted with insulin by exocytosis. Specific immunolocalization of D-alanine in pituitary ACTH-secreting cells and pancreatic ß-cells suggests that this amino acid participates in blood glucose regulation in mammals. By modulating insulin secretion, D-serine probably participates in the control of systemic glucose metabolism by modulating insulin secretion. We anticipate that future investigation will significantly increase the functional repertoire of D-amino acids in homeostatic control.

Accounting for ingrowth of radioactive progeny in dose assessments: generic weighting factors for dose coefficients.

In this paper, we describe a practical and convenient method to include the contribution of the ingrowth of radioactive progeny in dose assessments of the corresponding parent nuclides. This method modifies the dose coefficients (DCs) of parent nuclides by adding weighted DCs of the corresponding daughter nuclides to them. Based on the decay kinetics of serial nuclear transformations, the progeny weighting factors, with values between 0 and 1, are derived by analysis of the time-integrated activity of each nuclide in the (branched) decay chain headed by a parent nuclide. Using the electronic, nuclear-decay database of Publication 107 of the International Commission on Radiological Protection (ICRP 2008), DC weighting factors for annual dose assessments are calculated for all daughter radionuclides in the decay chains and are tabulated in this paper. Weighting factors based on integration periods other than one year, ranging from 1 h-70 years, are also provided (see the supplementary material). With a priori established weighting factors, dose assessments become significantly simplified by considering the decay kinetics of only the parent nuclides and by applying the modified DCs. This ensures that the ingrowth of progeny is taken into account realistically. In some cases, one requires a conservative estimate of the dose, for instance when dealing with issues of the clearance of materials under regulatory control. Therefore, we adapted the weighting-factor method to derive conservative DC weighting factors for dose evaluations. These values are calculated for various integration periods and compared with those from an existing method adopted by the Euratom Article 31 Group of Experts and by the International Atomic Energy Agency. Identical progeny weighting factors are obtained for long-lived parent radionuclides, whereas for short-lived parent radionuclides, the new method can yield significantly larger values. For example, the weighting factor of I-131 (daughter of parent Te-131) increases from 0.002 to 1.0 based on an integration period of 1 year. The progeny DC weighting factors, derived based on nuclear transformations in exit-only decay chains, may not always be suitable for use in radiological dose evaluations. For instance, when environmental removal pathways are dominant, the application of these weighting factors may have its limitations. This paper, therefore, provides guidance on the proper selection and application of weighting factors.

AMPA receptor expression in mouse testis and spermatogonial GC-1 cells: A study on its regulation by excitatory amino acids.

Excitatory amino acids (EAAs) are found present in the nervous and reproductive systems of animals. Numerous studies have demonstrated a regulatory role for Glutamate (Glu), d-aspartate ( d-Asp) and N-methyl- d-aspartate (NMDA) in the control of spermatogenesis. EAAs are able to stimulate the Glutamate receptors, including the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR). Here in, we assess expression of the main AMPAR subunits, GluA1 and GluA2/3, in the mouse testis and in spermatogonial GC-1 cells. The results showed that both GluA1 and GluA2/3 were localized in mouse testis prevalently in spermatogonia. The subunit GluA2/3 was more highly expressed compared with GluA1 in both the testis and the GC-1 cells. Subsequently, GC-1 cells were incubated with medium containing l-Glu, d-Glu, d-Asp or NMDA to determine GluA1 and GluA2/3 expressions. At 30 minutes and 2 hours of incubation, EAA-treated GC-1 cells showed significantly higher expression levels of both GluA1 and GluA2/3. Furthermore, p-extracellular signal-regulated kinase (ERK), p-Akt, proliferating cell nuclear antigen (PCNA), and Aurora B expressions were assayed in l-Glu-, d-Glu-, and NMDA-treated GC-1 cells. At 30 minutes and 2 hours of incubation, treated GC-1 cells showed significantly higher expression levels of p-ERK and p-Akt. A consequent increase of PCNA and Aurora B expressions was induced by l-Glu and NMDA, but not by d-Glu. Our study demonstrates a direct effect of the EAAs on spermatogonial activity. In addition, the increased protein expression levels of GluA1 and GluA2/3 in EAA-treated GC-1 cells suggest that EAAs could activate ERK and Akt pathways through the AMPAR. Finally, the increased PCNA and Aurora B levels may imply an enhanced proliferative activity.

Seasonal expression and cellular distribution of star and steroidogenic enzymes in quail testis.

The quail Coturnix coturnix is a seasonal breeder with a physiological switch on/off of gonadic activity. Photoperiod and temperature are the major environmental factors regulating the spermatogenesis. To more thoroughly comprehend the steroidogenic pathways that govern the seasonal reproductive cycle, we have investigated the localization of StAR protein and steroidogenic enzymes (3ß-HSD, 17ß-HSD, P450 aromatase, and 5α-Red) as well as androgen and estrogen levels, in the testis of reproductive and nonreproductive quails. We demonstrated that StAR, 3ß-HSD, 17ß-HSD, P450 aromatase, and 5α-Red were always present in the somatic (Leydig and Sertoli cells) and germ cells (spermatogonia, spermatocytes I and II, spermatids, and spermatozoa). In addition, by western blot analysis, we demonstrated that 17ß-HSD, P450 aromatase, and 5α-Red showed the highest expression levels during the reproductive testis compared with nonreproductive one. Accordingly, we also found that during the reproductive phase the highest titres of testosterone, 17ß-estradiol, and 5α-dihydrotestosterone are recorded. In conclusion, our findings demonstrated that in C. coturnix: (a) both somatic and germ cells are involved in the local synthesis of sex hormones; (b) 17ß-HSD, P450 aromatase, and 5α-Red expressions, as well as testicular androgens and estrogens, increased in reproductive quail testis. This study strongly indicates that the steroidogenic process in quail testis exhibits seasonal changes with the promotion of both androgenic and estrogenic pathways in the reproductive period, suggesting their synergic mechanism in the spermatogenesis regulation.

Filling the gap between chemical carcinogenesis and the hallmarks of cancer: A temporal perspective.

BACKGROUND: Cancer is believed to arise through the perturbation of pathways and the order of pathway perturbation events can enhance understanding and evaluation of carcinogenicity. This order has not been examined so far, and this study aimed to fill this gap by attempting to gather evidence on the potential temporal sequence of events in carcinogenesis. DESIGN: The methodology followed was to discuss first the temporal sequence of hallmarks of cancer from the point of view of pathological specimens of cancer (essentially branched mutations) and then to consider the hallmarks of cancer that one well-known carcinogen, benzo(a)pyrene, can modify. RESULTS: Even though the sequential order of driving genetic alterations can vary between and within tumours, the main cancer pathways affected are almost ubiquitous and follow a generally common sequence: resisting cell death, insensitivity to antigrowth signals, sustained proliferation, deregulated energetics, replicative immortality and activation of invasion and metastasis. The first 3 hallmarks can be regarded as almost simultaneous while angiogenesis and avoiding immune destruction are perhaps the only hallmarks with a varying position in the above sequence. CONCLUSIONS: Our review of hallmarks of cancer and their temporal sequence, based on mutational spectra in biopsies from different cancer sites, allowed us to propose a hypothetical temporal sequence of the hallmarks. This sequence can add molecular support to the evaluation of an agent as a carcinogen as it can be used as a conceptual framework for organising and evaluating the strength of existing evidence.

Causal narratives in public health: the difference between mechanisms of aetiology and mechanisms of prevention in non-communicable diseases.

Research in the health sciences has been highly successful in revealing the aetiologies of many morbidities, particularly those involving the microbiology of communicable disease. This success has helped form a narrative to be found in numerous public health documents, about interventions to reduce the burden of non-communicable diseases (e.g., obesity or alcohol related pathologies). These focus on tackling the purported pathogenic factors causing the diseases as a means of prevention. In this paper, we argue that this approach has been sub-optimal. The mechanisms of aetiology and of prevention are sometimes significantly different and failure to make this distinction has hindered efforts at preventing non-communicable diseases linked to diet, exercise and alcohol consumption. We propose a sociological approach as an alternative based on social practice theory. (A virtual abstract for this paper can be found at: https://www.youtube.com/channel/UC_979cmCmR9rLrKuD7z0ycA).

Causality in cancer research: a journey through models in molecular epidemiology and their philosophical interpretation.

In the last decades, Systems Biology (including cancer research) has been driven by technology, statistical modelling and bioinformatics. In this paper we try to bring biological and philosophical thinking back. We thus aim at making different traditions of thought compatible: (a) causality in epidemiology and in philosophical theorizing-notably, the "sufficient-component-cause framework" and the "mark transmission" approach; (b) new acquisitions about disease pathogenesis, e.g. the "branched model" in cancer, and the role of biomarkers in this process; (c) the burgeoning of omics research, with a large number of "signals" and of associations that need to be interpreted. In the paper we summarize first the current views on carcinogenesis, and then explore the relevance of current philosophical interpretations of "cancer causes". We try to offer a unifying framework to incorporate biomarkers and omic data into causal models, referring to a position called "evidential pluralism". According to this view, causal reasoning is based on both "evidence of difference-making" (e.g. associations) and on "evidence of underlying biological mechanisms". We conceptualize the way scientists detect and trace signals in terms of information transmission, which is a generalization of the mark transmission theory developed by philosopher Wesley Salmon. Our approach is capable of helping us conceptualize how heterogeneous factors such as micro and macro-biological and psycho-social-are causally linked. This is important not only to understand cancer etiology, but also to design public health policies that target the right causal factors at the macro-level.

The integration of social, behavioral, and biological mechanisms in models of pathogenesis.

A large part of contemporary medicine is concerned with describing and understanding the biological mechanisms involved in disease causation. Comparatively less attention has been paid to the socioeconomic and behavioral mechanisms underlying disease. This article argues for an integration of social, behavioral, and biological factors in the explanation of pathogenesis, a perspective that is in accord with the vision of pioneer public health practitioners of the 19th century, but that has gradually been overtaken by the dominance of the biomedical disease model. In recent decades, the social components of disease have been depicted as "distal" factors or used as "classificatory" devices. We explain how the integration we propose, which draws upon the concepts of "mixed mechanism" and of "lifeworld," advances the view of several scholars of the recent past. Finally, we discuss new findings in epigenetics and psychology, where socioeconomic disparities appear to be an integral part of the explanation of health conditions, to illustrate how the integration may work in practice.

The evidence that evidence-based medicine omits.

According to current hierarchies of evidence for EBM, evidence of correlation (e.g., from RCTs) is always more important than evidence of mechanisms when evaluating and establishing causal claims. We argue that evidence of mechanisms needs to be treated alongside evidence of correlation. This is for three reasons. First, correlation is always a fallible indicator of causation, subject in particular to the problem of confounding; evidence of mechanisms can in some cases be more important than evidence of correlation when assessing a causal claim. Second, evidence of mechanisms is often required in order to obtain evidence of correlation (for example, in order to set up and evaluate RCTs). Third, evidence of mechanisms is often required in order to generalise and apply causal claims. While the EBM movement has been enormously successful in making explicit and critically examining one aspect of our evidential practice, i.e., evidence of correlation, we wish to extend this line of work to make explicit and critically examine a second aspect of our evidential practices: evidence of mechanisms.

Navigating complex trade-offs in public health interventions.

Public health, just as any policy-related field, faces the evergreen problem of turning knowledge into action. Among other problems, there is a clash between the inherent complexity of public health problems and the inevitable push, by decision-makers and the public, to simplify them. The Covid-19 pandemic has shown the insufficiencies of our current epistemological, methodological and normative apparatus to handle such crises in a timely manner. Despite this, several authors have been arguing for the importance of engaging global crises such as Covid-19 in ways that do not oversimplify key dimensions of the issues involved. In this paper, we contribute to this emerging scholarship. Building on existing work in the field of environmental problem-solving, we propose an integrative approach to navigating complex trade-offs in public health interventions. Briefly put, we propose that decision making should be informed by an analysis of any given problem from four distinct, but interrelated, lenses: (i) values and valuation, (ii) process and governance, (iii) power and inequalities and (iv) scientific evidence, methods and concepts. This normative framework, we argue, can help with spelling out the complexity of public health problems and with spelling out the rationale behind public health decision making to non-specialists and the general public. We illustrate our approach using the controversy over wearing face masks in the Covid-19 pandemic.

The 'lifeworld' of health and disease and the design of public health interventions.

The consequences of the COVID-19 pandemic are still working through health systems worldwide, and further reflections about the nature of health and disease, and about how to design and implement effective public health interventions are much needed. For numerous diseases and conditions, as well as for COVID-19, our knowledge base is rich. We know a lot about the biology of the disease, and we have plenty of statistics that relate health to socio-economic factors. In this paper, we argue that we need to add a third dimension to this knowledge base, namely a thorough description of the lifeworld of health and disease, in terms of the mixed biosocial mechanisms that operate in it. We present the concepts of lifeworld and of mixed mechanisms, and then illustrate how they can be operationalised and measured through mixed methodologies that combine qualitative and quantitative approaches. Finally, we explain the complementarity of our approach with the biological and statistical dimensions of health and disease for the design of public health interventions.

Promoting the health of vulnerable populations: Three steps towards a systems-based re-orientation of public health intervention research.

This paper proposes a novel framework for the development of interventions in vulnerable populations. The framework combines a complex systems lens with syndemic theory. Whereas funding bodies, research organizations and reporting guidelines tend to encourage intervention research that (i) focuses on singular and predefined health outcomes, (ii) searches for generalizable cause-effect relationships, and (iii) aims to identify universally effective interventions, the paper suggests that a different direction is needed for addressing health inequities: We need to (i) start with exploratory analysis of population-level data, and (ii) invest in contextualized in-depth knowledge of the complex dynamics that produce health inequities in specific populations and settings, while we (iii) work with stakeholders at multiple levels to create change within systems.

Public health policy, evidence, and causation: lessons from the studies on obesity.

The paper addresses the question of how different types of evidence ought to inform public health policy. By analysing case studies on obesity, the paper draws lessons about the different roles that different types of evidence play in setting up public health policies. More specifically, it is argued that evidence of difference-making supports considerations about 'what works for whom in what circumstances', and that evidence of mechanisms provides information about the 'causal pathways' to intervene upon.