Fungi are eukaryotic heterotrophs present in virtually every environment, with potentially more than 5 million individual species . DespiteThe constant exposure of humans to both commensal and environmental fungi requires a competent immune system for tolerance, protection, and/or clearance, while limiting collateral damage caused by excessive or detrimental inflammation. Innate responses to fungal pathogens are initiated by fungal component recognition via an array of pathogen-associated molecular pattern recognition receptors (PRRs) –11. RecoIncreased understanding of these host-fungal interactions and the mechanisms that favor protective immunity over immune pathology could provide targets for novel therapeutic approaches that complement the limited repertoire of existing antifungal drugs. The aim of this research topic is to explore the host and fungal pathways that program innate and adaptive immunity and the immune cells, molecules, and regulatory pathways that comprise protective or detrimental responses to fungal exposure or infection. Researchers from 15 countries in North and South America, Europe, Asia, and Australia contributed 36 review and original research articles that cover a wide range of fungal pathogens, disease models, and effector and regulatory cell and molecular pathways of host immune responses to fungal exposure and infection. Here, we present an outline of the findings, perspectives, and reviews contained in this research topic.Tang et al.. Although the important role of the β-1,3-glucan receptor dectin-1 in antifungal immunity is well appreciated, the α-mannan-binding dectin-2 and dectin-3 also influence responses to fungal exposure and infection and their roles are less clear. In support of these studies, Preite et al. show that dectin-2 and dectin-3 mediate antifungal activity and cytokine secretion of human plasmacytoid dendritic cells (pDCs) in response to the endemic dimorphic fungal pathogen Paracoccidioides brasiliensis via a pathway mediated by the CLR-associated signaling molecule Syk. In addition, de Castro et al. show that mouse bone marrow-derived macrophages (BMMs) and DCs (BMDCs) produce IL-1β and IL-18 in response to Fonsecaea pedrosoi, the main causative agent of chromoblastomycosis, in a dectin−1, −2, and −3-dependent manner. A role for the NOD-Like Receptor P3 (NLRP3), an intracellular protein complex that controls activation of inflammatory caspases and cytokine production, is shown by Feriotti et al. as important for the development of protective immunity to pulmonary infection with P. brasiliensis. β-glucan stimulation of NLRP3 inflammasome-mediated IL-1β secretion in B cells reported by Ali et al. shows innate antifungal cytokine production in an adaptive lymphocytes that were also capable of producing IgM in an NLRP3-dependent manner.The major class of pattern recognition receptors, the C-type lectin receptors (CLRs), including molecules critical for the initiation of inflammation and the development of adaptive immunity to fungi, are reviewed by Aspergillus fumigatus . Circulating monocytes are subsequently recruited to sites of infection in response to inflammatory signals produced by tissue-resident macrophages. Tóth et al. report that the exposure of hyphae of the dematiaceous mold Curvularia lunata to human THP-1 monocytes resulted in increased inflammatory IL-8 and regulatory/anti-inflammatory IL-10, suggesting a possible mechanism for the ability of this species to cause chronic infections in immune competent individuals. Landgraf et al. report that dihydrolipoyl dehydrogenase secreted by P. brasiliensis may act as an exoantigen that enhances macrophage phagocytosis. Mukaremera et al. report that hypha of the yeast Candida albicans induced low levels of cytokine secretion from human monocytes, with the highest levels from yeast and intermediate levels from pseudohypha, and cell wall mannan depletion partially reversed these responses. Using fluorescence yeast cell wall-labeling to measure yeast division within macrophages, Dagher et al. show that activation of the tyrosine kinase Syk is critical for macrophage control of phagocytosed C. glabrata. Together, these studies provide new insights into the roles for monocytes and macrophages in induction and regulation of antifungal inflammation and fungal clearance.PRR-expressing tissue-resident macrophages are part of the first line of defense against fungal infections  cells in innate antifungal immunity is reviewed by Schmidt et al.. These works further illustrate diverse mechanisms and roles for innate myeloid and lymphoid cells in antifungal immunity.Inflammatory cytokine production by macrophages leads to localized inflammation with production of chemokines that attract additional innate immune cells. Maldonado and Fitzgerald-Bocarsly. Xu et al. investigated the role of macrophage receptor with collagenous structure (MARCO) on pDC recruitment and induction of antifungal adaptive immunity to C. neoformans infection, and report that expression of MARCO promoted recruitment of lymph node DCs and skewed local and systemic T helper cell responses away from protective Th1 responses and toward non-protective Th2 responses. Wong et al. demonstrate a role for leucine-rich repeat kinase 2 (LRRK2) in translocation of NFAT to the nucleus in DCs in the early stages of the non-canonical autophagic response to A. fumigatus conidia, thus connecting LRRK2 with NFAT-mediated IL-2 expression in early antifungal immunity. Hellmann et al. compared immune responses to A. fumigatus in human and mouse DCs, macrophages, and neutrophils, and observed that human DCs exhibited more significant increases in maturation markers and phagocytic ability than murine DCs, while murine neutrophils and macrophages displayed more reactive oxygen species production after A. fumigatus exposure. Collectively, these works provide support for additional mediators of DC function that shape antifungal adaptive immunity, and suggest distinct roles for these cells in human fungal disease and experimental animal models.Dendritic cells (DCs) are multifaceted professional antigen presenting cells that integrate antigen uptake and local inflammatory signals in order to effectively prime adaptive antifungal immune responses upon migration to site-draining lymph nodes . The rolTristão et al. report that these cytokines, along with the IL-17 receptor A, were necessary for protective lung granuloma formation in mice infected with P. brasiliensis. IL-1α/β also influence adaptive immune responses to fungi, such as the dimorphic Cryptococcus spp., as Shourian et al. observed that IL-1 receptor type 1-deficient mice had increased cryptococcal burden, eosinophilia, M2 macrophage polarization, and recruitment of CD4+IL-13+ T cells, with a concomitant reduction in pro-inflammatory Th1 and Th17 cytokines. However, excessive inflammation and Th1 activation in C. neoformans infection is also detrimental, and was reported by Fa et al. to be mediated by the cell death regulatory genes FADD (Fas-associated death domain) and RIP3K (receptor interacting protein kinase 3). The results of these studies support roles for Th1 and Th17 responses in antifungal immunity, as well as a requirement for regulation of cell death to limit excessive inflammation.After DCs migrate to draining lymph nodes, they present fungal antigen to naïve T cells, inducing antifungal adaptive immunity. This process is influenced by cytokines in the microenvironment. Formation of the antifungal Th17 subset of T helper cells is promoted by the inflammatory cytokines IL-6 and IL-23, and A. fumigatus; findings that shed light on this host-pathogen dynamic are reviewed by Choera et al.. De Araújo et al. report that DCs from an infected P. brasiliensis-susceptible mouse strain exhibit IDO activity and promote inflammation, while DCs from resistant mice phosphorylate IDO and promote a tolerogenic phenotype. The same group further compared P. brasiliensis infection in IDO-deficient and wild-type mice, and observed increased mortality, fungal burden, histopathology, and Th17 cell recruitment and activation in IDO-deficient mice with concomitant decreases in Th1 and Treg cells de Araújo et al.. In addition to the enzymatic activity and signaling by IDO, antifungal immune responses are regulated at the post-transcriptional level by microRNAs (miRNAs); these molecules and their associated pathways are reviewed by Croston et al.. Zimmerman et al. report that patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) exhibit decreased levels of the signaling molecule STAT1, in contrast with patients with STAT1 gain-of-function mutations, despite similar disease phenotypes. These works highlight findings of enzymatic, post-translational, and signal transduction in the regulation of antifungal immunity and fungal disease susceptibility.Cell and molecular regulation of immunity occurs at multiple levels from induction to resolution of immune responses. An enzyme that limits available tryptophan and thus dampens immune cell proliferation and effector function, indoleamine 2,3-dioxygenase (IDO), is produced by mammalian hosts and the fungal pathogen Kernien et al. review host immune recognition of fungal biofilms and biofilm properties that inhibit host recognition and clearance. Zhang et al. explore how immune interactions with respiratory and gastrointestinal fungal microbiota contribute to chronic airway inflammatory disease. Sparber and LeibundGut-Landmann more specifically discuss host immune responses to skin-colonizing Malassezia species, yeasts that are involved in a variety of skin disorders. Finally, Casadevall et al. review the evidence that C. neoformans mediates host damage at the molecular, cellular, tissue, and organism levels, in some instances with formation of large fungal masses in brain tissue. These reviews highlight emerging areas of fungal immunology that consider fungal interactions with the host immune system beyond the level of cell and molecular recognition.Although spore/hyphal recognition by innate cells drives the development of immunity to fungal pathogens, host-fungal pathogen interactions are also considered at the macroscopic level of microbial communities  and host tissues. Elsegeiny et al. review studies of immune pathology in cryptococcal infection and discuss the need for immune targeting therapies that suppress immune-mediated damage without further compromising host protection. Van Dyke et al. demonstrate that combining non-lethal C. neoformans challenge with local IFN-γ production elicits an immune response that protects mice from subsequent lethal infection. Tso et al. summarize efforts and obstacles in the development of anti-Candida vaccines, and discuss the potential use of trained immunity in the development of strategies against opportunistic fungal infections. Finally, Kumaresan et al. review the development of CD8+ T cell therapy for the control of invasive fungal infections, with a focus on the use of chimeric antigen receptor (CAR) T cells that target β-glucan. These reviews underscore the importance of current and future efforts to enhance immune protection while limiting immune pathology in patients that may not respond to traditional antifungal therapies.Despite significant advances in our understanding of host immunity to fungal exposure and infection, treatment of fungal diseases has not progressed beyond the use of a limited repertoire of antifungal drugs that are rendered increasingly ineffective by emerging fungal resistance. Since appropriate antifungal immunity is critical for protection from fungal disease, complementary therapies that target immune pathways represent areas of considerable interest for fungal immunologists. Collectively, the studies described in original research and review articles in this topic provide optimism for the future of antifungal immune therapy. Recent advances by fungal immunologists have greatly increased our understanding of the basic mechanisms of innate immune recognition, inflammation, adaptive immunity, and regulation of antifungal immune responses at molecular, cell, tissue, and organismal levels. We hope these articles will stimulate further research with the ultimate goal of improving outcomes for patients with fungal diseases.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.


People with diabetes mellitus (DM) have a higher-than-average risk of having a heart attack or stroke , 2. In fDe Rosa et al., from Magna Graecia University, elegantly illustrated fundamental genetic and epigenetic mechanisms linking cardiovascular disease and DM; similarly, Pordzik et al. identified the functional role of specific platelet-related microRNAs in the pathophysiology of cardiovascular events in high-risk populations, including diabetic patients.Soares Felicio et al. demonstrated an association between reduced levels of Vitamin D and the presence and severity of diabetic kidney disease in type 1 DM (T1DM); the molecular mechanisms underlying diabetic nephropathy have been also explored by Zou et al. in streptozotocin-induced DM. Arcangeli et al. found a significant association between the number of circulating endothelial progenitor cells (cEPCs) and the age and duration of the disease in T1DM patients: indeed, young T1DM patients have significantly higher levels of cEPCs compared to adult T1DM patients; of note, such difference is also maintained when the disease lasts for more than 10 years. The Authors propose that maintaining a high number of cEPCs, possibly through an efficient glycemic control, would contribute to contain the cardiovascular burden in T1DM. Notably, in vitro experiments performed by Lin et al. at New York University have shown how to ameliorate purification and maturation of human induced pluripotent stem cell (iPSC)-derived cardiomyocytes through means of culture in glucose-depleted medium supplemented with fatty acids (oleic acid and linoleic acid) and 3,3′,5-triiodo-L-thyronine (T3).Infante et al. revealed a greater severity of coronary artery disease in type 2 diabetes (T2DM) patients compared to non-diabetic individuals; equally important, van Bussel et al. from Maastricht University Medical Center, highlighted the actual advantages of multiparametric neuroimaging in the clinical evaluation of cognitive decline in T2DM.Applying comprehensive analyses based on imaging and molecular biology, Orosz et al. in subjects with impaired glucose tolerance, a prediabetic condition, have shown that prediabetes is associated with repolarization instability, indicated by elevated values of beat-to-beat short-term QT interval variability, thereby suggesting that an impaired autonomic control precedes the actual onset of diabetes. Last but not least, Altara et al. validated the key importance of targeting microvascular disease, common in both diabetes and obesity, in order to treat heart failure with preserved ejection fraction (HFpEF). Microvascular disease is a growing public health problem, accounting for approximately half of hospital admissions of individuals with heart failure (The studies performed by  failure , 11, 12.In summary, the present Research Topic indicates that the exceptional advances achieved in the last decade in understanding the molecular alterations involved in the pathophysiology of both DM and cardiovascular disease are opening new therapeutic opportunities for the treatment of these disorders and, potentially, their future application to the clinical scenario might result to further enhancements in patient care. Furthermore, the exciting findings discussed herein might foster community awareness of these important diseases and stimulate further research in the field.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Numerous studies have shown that infiltration of lymphocytes into tumor tissue is associated with better prognosis (Bindea et al., 2013; SchmidtThis result is of relevance, because it shows that immune modifiers have different roles in NSCLC of smokers and non-smokers. It will be important to consider this difference in therapy studies targeting the PD-1/PD-L1 axis. Predicting prognosis and response to therapy remains a challenging task (Hellwig et al., 2016; Lohr etThe author declares no conflict of interest.


Cervico-vaginal mucus (CVM), the product of epithelial cells lining the uterus, cervix and vagina, is secreted to facilitate uterine lubrication and microbial clearance. Predominantly composed of water and mucins, CVM also contains high levels of immuno-active proteins such as immunoglobulin A (IgA), lactoferrin and lysozyme which protect against infection by blocking adhesion and mediating microbial killing. The repertoire of cytokines, chemokines and antimicrobial peptides is predominantly generated by the secretions of endometrial epithelial cells into the uterine lumen and concentrated in the CVM. The quantity and relative proportions of these inflammatory biomarkers are affected by diverse factors including the estrus cycle and health status of the animal and therefore potentially provide important diagnostic and prognostic indicators. We propose that measuring molecular signatures in bovine CVM could be a useful approach to identifying and monitoring genital tract pathologies in beef and dairy cows. Cervico-vaginal mucus (CVM) represents a mixture of vaginal, cervical and uterine mucus and is composed of 92–95% of water, ions and 5–8% solid matter   multiplication in the uterus  are secreted directly into the endometrial lumen while TNF-α, IL-1 and IL-8 are concentrated in uterine mucus after infiltrating through uterine wall (Carneiro et al. IL8 in the endometrium 2 weeks after calving, and intrauterine infusion of exogenous IL-8 reproduces the disease (Chapwanya et al. IL-1 is the key mediator of uterine inflammation that is secreted as a result of tissue damage associated with parturition (Adnane et al. E. coli infection (Chapwanya et al. E. coli have high systemic levels of AGP at 21 and 28 DPP (Williams et al. Trichomonas foetus or endometritis (Adnane et al. APPs are highly responsive proteins, secreted by the liver in response to injury or infection in an effort to restore homeostasis. Extra-hepatic sources of APPs have also been identified, including in endometrial cells (Chapwanya et al. Members of the S100 family chelate calcium and therefore regulate many cellular processes including microbial viability (Corbin et al. Here we reviewed the measurement of inflammatory biomarkers in CVM early postpartum as an alternative method for the prognosis of genital tract pathology in cows and other species. CVM molecular signatures are specific to uterine health status may also reflect local microbiome diversity. They provide a convenient, cost effective and welfare-friendly method for timely detection of uterine inflammation in order to reduce the impact of endometritis on dairy cow production. CVM therefore is a potentially valuable resource to investigate the diversity of bacteria in cows with uterine disease as many of these bacteria are trapped by the mucus structure.


DiMarco et al., Chem. Sci., 2018, DOI: 10.1039/c7sc03839a.Correction for ‘Dye-sensitized electron transfer from TiO The authors regret that The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.


Microglia are the resident immune cells of the central nervous system and have many functions including neuroinflammation, axonal guidance and neurotrophic support. We aimed to provide a quantitative review of g). The difference in 18-kDa translocator protein (TSPO) binding between patients with schizophrenia and healthy controls, as quantified by either binding potential (BP) or volume of distribution (VT), was used as the main outcome. Sub-analysis and sensitivity analysis were carried out to investigate the effects of genotype, ligand and illness stage.Demographic, clinical and imaging measures were extracted from each study and meta-analysis was conducted using a random-effects model  but no significant differences when VT was used .In total, 12 studies comprising 190 patients with schizophrenia and 200 healthy controls met inclusion criteria. There was a significant elevation in tracer binding in schizophrenia patients relative to controls when BP was used as an outcome measure, (Hedge's In conclusion, there is evidence for moderate elevations in TSPO tracer binding in grey matter relative to other brain tissue in schizophrenia when using BP as an outcome measure, but no difference when VT is the outcome measure. We discuss the relevance of these findings as well as the methodological issues that may underlie the contrasting difference between these outcomes. However, findings have been inconsistent and so far they have only been partially reviewed quantitatively . In order to extract data from studies where data were available only in a plot format, we have used the plot digitiser software (http://plotdigitizer.sourceforge.net/).The main outcome measure was the difference in the TSPO imaging index between patients with schizophrenia-spectrum disorders and healthy controls. For all studies, we extracted the following variables: authors, year of publication, subject characteristics for the patient and healthy control group , imaging characteristics  and modelling method. The estimation of pooled standard deviation was performed using the statstodo software  . The DVR method used in this study is not equivalent to the measures used in the other studies. Also Ottoy et al.  . Therefore, the main analysis of VT values included six studies using the 2TCM model.The main outcome measure was the effect size determined using the TSPO tracer measure and quantified by either BPd Hickie , and alt  express both mixed affinity for TSPO (MAB), while those without the polymorphism (HH) have high-affinity binding (HAB) for TSPO . Leave-one-out sensitivity analyses were conducted to investigate the potential effect of an individual study on the outcome measure. A p value <0.05 (two-tailed) was taken as a significance level. The statistical analysis of the extracted data was conducted using the R statistical programming language version 3.2.2 with the ‘metafor’ package.Publication bias was assessed using visual inspection of funnel plots as well as regression test. Where potential publication bias was suspected, trim and fill analysis was conducted to correct for putatively missing studies. Heterogeneity was estimated using the The PubMed, EMBASE and PsycINFO databases were searched without language restrictions. The electronic search using EMBASE and PsycINFO were carried out together using Ovid. The following keywords were used: ‘(Positron Emission Tomography OR PET OR Single photon emission tomography OR SPET OR Single Photon Emission Computed Tomography OR SPECT) AND (schizophrenia OR schizophreniform OR psychosis) AND ’. Review papers were also screened to search for additional studies.The inclusion criteria were: original studies in (1) patients with a diagnosis of schizophrenia or related psychotic diagnoses , (2) reporting PET measures using a TSPO-specific ligand and (3) reporting data for the whole grey matter or grey matter regions. Studies that did not have a control group were excluded. Where there was sample overlap between studies, we included the largest one and excluded the other to avoid double counting.VT). As these give different information, we conducted separate meta-analyses of these outcomes. The studies that used BP as an outcome measure have used either values obtained using microparameters from Simplified Reference Tissue Model  or volume of distribution  0.02–0.6] . Visual inspection of the funnel plot suggested asymmetry . The regression test was significant . The trim and fill analysis showed three putatively missing studies on the left side. The results were not significant after correcting for these studies . The results were significant in two out of six in the leave-one-out analysis, with effect sizes varying from 0.27 to 0.47. Five out of the six studies used the [11C]-PK11195 ligand, with the sub-analysis of these studies revealing a significant elevation of BP in patients with schizophrenia with an effect size of 0.35  .Six studies reported outcome measures as BP. Our results showed that BP was significantly elevated in patients with schizophrenia when compared with healthy controls with an effect size of 0.31 [Hedge's .02–0.6] . The I2 VT. VT in patients with schizophrenia when compared with healthy controls . The I2 test revealed moderate–high heterogeneity . Visual inspection of funnel plot suggested asymmetry . However, the regression test was not significant . Trim and fill analysis showed two missing studies in the right side. The effect sizes varied from −0.08 to −0.37 in the leave-one-out analysis. We extracted data for high- (HABs) and mixed-affinity binders (MABs) separately from these studies to conduct a sub-analysis stratified by genotype. Bloomfield et al. reported only one patient who was a MAB, precluding accurate estimation of the effect size. Thus, this study was not included in the sub-analysis of MAB subjects . However, there was a significant difference in the MAB .Six studies reported the outcome measure as g = 0.31; p = 0.03), but there is no significant difference when the tracer volume of distribution (VT) is used as the outcome measure . In the following section, we consider methodological factors and the implications of our findings.Our main findings are that TSPO PET tracer binding is significantly elevated in patients with schizophrenia relative to controls when BP is used as an outcome measure, with a small-to-moderate effect size  is negative, the results using this tracer are not an outlier, suggesting findings may not entirely be accounted for by tracer differences. It has been suggested that TSPO expression may change during the course of the disorder, which could account for the differences in findings between studies  measures the tracer binding in the tissue of interest relative to another brain region selected to have negligible specific binding, to give specific binding in the region of interest  measures the total amount of tracer in the brain region relative to that in the blood  that are known to be elevated in schizophrenia (Telford et al., et al., et al., VT, and potentially mask an elevation in the patients’ brain as compared with controls. There is inconsistency in findings in schizophrenia, with one study showing elevation in the plasma concentration levels of [11C]-PBR28 in patients with schizophrenia relative to controls (Bloomfield et al., 18F]-FEPPA and [11C]-PBR28 show no differences in plasma concentration levels (Hafizi et al., et al., et al., et al., et al., VT values in the disorder, but it should be recognized that there is no direct evidence that plasma binding alters VT values in humans (Cumming et al., VT changes are not explained by an effect of protein binding (Collste et al., VT in schizophrenia. Nevertheless, while this is a potential concern for studies using VT as the outcome measure, blood binding should not affect the studies that use a ratio approach, as these methods report tracer uptake relative to another brain region rather than blood. Furthermore, TSPO is expressed on the endothelial cells of brain blood vessels as well as on the outer layer of mitochondria in microglia (Rizzo et al., et al., The non-displaceable BP (BPet al., et al., et al., et al., et al., Taken together, our meta-analytic findings suggest an elevation in TSPO tracer binding in total grey matter relative to other brain tissue, but not relative to blood, with the caveat that the relative increase is largely based on studies using PK11195, which has a lower specific signal. Thus, this could reflect an increase in TSPO in grey matter or a reduction in TSPO in the reference region, or altered non-specific binding in the brain (Cumming VT as the outcome measure, showed a reduction in VT in patients relative to healthy controls (Plaven-Sigray et al., VT in our meta-analysis, our meta-analysis included one more second-generation study showing no differences in VT between schizophrenia patients and healthy controls (Ottoy et al., in utero and early development may predispose to schizophrenia (Meyer, VT approach (Bloomfield et al., VT as the outcome measure, using the tracer [18F]-FEPPA and [11C]-PK11195, showed no differences in TSPO binding between healthy controls and in individuals at ultra high risk of psychosis for psychosis (Hafizi et al., et al., et al., et al. (et al. (et al. (Our different findings depending on the outcome measure used point towards the existence of potential methodological problems in TSPO imaging, raising questions over the interpretation of the elevation in grey matter TSPO binding relative to a reference region in patients with schizophrenia when compared with healthy controls. A recent individual participant data meta-analysis of second-generation radioligand studies, with a Meyer, . However, et al.  and van  (et al.  suggest  (et al. b using tVT is the outcome measure used. These results suggest that potential methodological differences between TSPO studies need to be accounted for and addressed in future studies and keep open the discussion over the existence of an increase in microglia activity in patients with schizophrenia-spectrum disorders.In conclusion, there is evidence for a moderate effect size elevation in TSPO tracer binding in grey matter in schizophrenia-spectrum disorders when using BP as an outcome measure, but no changes when


Purified proteins offer a homogeneous population of biological nanoparticles, equipped in many cases with specific binding sites enabling the directed self-assembly of envisaged one-, two- or three-dimensional arrays. These arrays may serve as nanoscale biotemplates for the preparation of novel functional composite materials, which exhibit potential applications, especially in the fields of nanoelectronics and optical devices. This review provides an overview of the field of protein-mediated biotemplating, focussing on achievements made throughout the past decade. It is comprised of seven sections designed according to the size and configuration of the protein-made biotemplate. Each section describes the design and size of the biotemplate, the resulting hybrid structures, the fabrication methodology, the analytical tools employed for the structural analysis of the hybrids obtained, and, finally, their claimed/intended applications and a feasibility demonstration (whenever available). In conclusion, a short assessment of the overall status of the achievements already made vs. the future challenges of this field is provided. Purified proteins offer a homogeneous population of biological nanoparticles, equipped in many cases with specific binding sites enabling directed self-assembly into one-, two- or three-dimensional arrays. These arrays may serve as biotemplates for the preparation of novel functional composite materials on the nanoscale, using either specific or site-directed chemical binding, with the potential for expansion by genetic or chemical manipulations.The potential inherent in protein-mediated biotemplating has attracted much attention throughout the past two decades . SeveraThis review provides an overview of the field of protein-mediated nanoscale biotemplating, focused on the achievements made throughout the past decade. The review is organized into sections according to the size and configuration of the protein-made biotemplate ranging from single protein soluble molecules and supramolecular protein “cages” to two-dimensional (2D) protein arrays, protein fibers and tubes, viral envelopes, three-dimensional (3D) protein crystals, and fragments of microbial cells.Each section describes the structure and size of the biotemplates employed, the fabrication methodology, the analytical tools used for its structural analysis, its claimed/intended applications and a feasibility demonstration (whenever available).Dagan-Moscovich et al.  initiateThe construction of a conducting metallic silver coating on the surface of the oxidoreductase enzyme glucose oxidase (GOx), a 5 × 8 nm ‘peanut-like’ dimer, enabled the nanowiring of its active site to platinum electrodes, resulting in the oxygen-independent direct determination of glucose by electrochemical biosensors , as scheThe biotemplating approach for electroless metal deposition directed to the surface of single soluble protein molecules was readily expanded from silver deposition to the deposition of other metals . GlucoseBy changing the biologically active protein core from GOx to the binding protein avidin, which is capable of specifically binding the vitamin biotin, additional applications of metallic silver-coated soluble protein molecules became available: imaging by HRTEM without staining. Mor et al.  demonstrThe range of applications of silver–enzyme hybrids was expanded further to antimicrobial applications aiming at the prevention of microbial biofilm formation, by affecting the on-site enzymatically attenuated release of silver ions, generating in situ antimicrobial activity. ‘Zone of inhibition’ antimicrobial studies demonstrated the feasibility of using these hybrids as wide-spectrum antimicrobial agents, especially as potential antifungal agents against skin and hair fungal contaminations .Veelders and Essen  demonstr2S nanoparticles (NPs) ~20 nm in diameter, enabled by RNase A capping, as hollow or full spheres with a claimed potential application as a drug delivery vehicle. The structural characterization of the QDs and Ag2S NPs thus obtained was carried out by high-resolution electron microscopy and spectral analysis.Kong et al.  describeThree recent reports have described biomineralization processes directed by protein molecules and designed for biomedical applications, including (i) the formation of albumin-mediated platinum nanocrystals for tomography imaging ; (ii) siFerritin—a natural, spherical, protein-made nanoscale “cage”—serves as an iron storage protein in bacteria, plants, animals and humans. In eukaryotic cells, ferritin is comprised of 24 subunits forming a hollow spherical shell with an inner diameter of 8 nm ,18). TheThe currently available biochemical data on the molecular mechanisms involved in these processes has inspired an exploration of the potential biomedical applications of a series of ferritin and mutated ferritin as protein cages for the preparation of hybrids by biotemplating. The templating of metals, alloys and metal ion salts, including iron, silver, gold, palladium, cadmium, cobalt, platinum and copper, were described . It Listeria innocua [The demonstrated feasibility of using the cavity, 7 nm in diameter, of natural ferritin for biotemplating has triggered efforts to develop analogous, smaller protein cages made of other proteins, resulting in CdS and CdSe NPs biotemplated inside the 4.5 nm diameter cavity of a cage-shaped protein derived from  innocua ,24, as wThe use of the cage-forming protein clathrin as a biotemplate for the synthesis of metallic NPs was also described, enabling the preparation of either homogeneous or bimetallic silver–gold or silver–copper NPs, albeit in sizes larger than those available by ferritin-mediated biotemplating (characterized by HRTEM) .2 molecules by using a small de novo designed protein, self-assembled as a “nanoreactor” for biomineralization. Behrens et al. [Voet et al.  demonstrs et al.  describeZhou et al.  describeThe pioneering work of Slytr et al.  on the iThe elucidation of the mechanism underlying the biotemplating of FePt NPs from the gas phase into a 2D array on the surface of a bacterial S-layer was presented by Queitsch et al.  and veriValero et al.  demonstrTwo-dimensional protein arrays other than bacterial S-layers were also used as biotemplates: Shindel and coworkers ,38,39 ex2 nanotubes enabled directed gold deposition, resulting in a hybrid exhibiting enhanced electron transfer between heme proteins and electrodes [Bovine serum albumin adsorbed as a 2D monolayer on the surface of TiOectrodes .Magnetospirillum magneticum for biotemplating of magnetic nanoparticle arrays; characterized by TEM, SEM and AFM, with potential applications in the manufacture of electric circuits.Galloway et al.  and BirdThe data available in the literature on self-assembly of proteins into fibers and tubes has paved the way for the exploitation of the inherent potential of using a series of readily available self-assembled protein fibers as biotemplates.Colby et al.  describeMalisauskas et al.  describeMethanocaldococcus jannaschii as a biotemplate decorated with silver, gold, palladium or platinum NPs, by electroless deposition using NaBH4 as a reducer of pre-adsorbed metal ions. The coated protein filaments thus obtained were characterized by TEM, spectral and electrical analysis, with the intended application of metal nanowires of defined length.Slocik et al.  describeIonov et al.  describep-nitrophenol to p-aminophenol by NaBH4. The gold nanoparticle-decorated lysozyme fibrils obtained were subjected to structural analysis by TEM and X-ray diffraction (XRD) analysis, and the feasibility of their use as a catalyst was demonstrated by kinetic spectrophotometric analysis.Juarez et al.  describeLeroux et al.  demonstr3O4–protein hybrids thus obtained were characterized by HRTEM, XRD and Fourier transform infrared spectroscopy (FTIR) analysis, and the feasibility of their magnetization demonstrated, leading towards potential applications as MRI contrast agents.The preparation of magnetic nanowires was also described by Juarez et al. . The coaGeobacillus stearothermophilus into tubular structures serving as biotemplates for the chemical deposition of platinum. The resulting hybrid was characterized by HRTEM, SEM and fluorescence analysis, in preparation for an intended application as nanowires.Korkmaz et al.  describe2 and TiO2 nanowires were grown on the biotemplate by monomer polymerization. The structures obtained were characterized by TEM and field-emission scanning electron microscopy (FESEM), and the feasibility of their use for biosensors based on the enzymatic activity of acetylcholine esterase was demonstrated and characterized.Padalkar et al.  describep-nitrophenol by NaBH4 to 4-aminophenol was described by Batzli and Love [Insulin fibers were coated with palladium by electroless deposition. The nanostructure and electrical properties of the palladium hybrids thus obtained were characterized by AFM, TEM, X-ray photoelectron spectroscopy (XPS) and scanning capacitance microscopy (SCM) analysis . The useand Love ; see alsand Love .Methanocaldococcus jannaschii, and its use as the major component of a biotemplate has been constructed in combination with other protein-made connecting units .The highly-ordered 3D protein envelopes of viruses, especially of plant viruses and bacteriophages, offer a ‘natural’ biotemplating tool, which is available in several geometrical shapes and sizes , used as a biotemplate for electroless deposition mediated by reduction with NaBH4 of cobalt and platinum ions within the channel. Nanowires ranging from 50 to 100 nm were identified by HRTEM analysis, as well as by EDXS and magnetometry measurements.Tsukamoto et al.  describe2 nanocrystals biotemplated onto the surface of M13 bacteriophages, significantly improving parameters such as operational and storage stability.Neltner et al.  describeLim et al.  describeThe production of 3 nm aligned magnetic NPs within the central channel of mutated tobamovirus was described by Kobayashi et al. . Energy-Nuraje et al.  demonstrThe controlled preparation of a 3D super lattice of metallic nanocrystals was demonstrated by Alloyeau et al. , combiniZhou et al.  describeFlexible electroactive nanomaterials were prepared by Vera-Robles et al.  by biote2+ cations [Piezoelectric nanowires were biotemplated on a genetically engineered M13 bacteriophage coating protein surface, displaying triglutamate residues for the periodical positioning of Pb cations . A latti cations  and tuna cations .As the structural complexity degree of biotemplated arrays grows, it should be mentioned that new characterization tools are also gradually becoming available for the biotemplating arena, as demonstrated by the recent report by Carreno-Fuentes et al.  on the cProtein crystals, routinely prepared for the elucidation of protein 3D structures by X-ray crystallography, present a highly ordered 3D array of protein molecules. Along with the formation of this array, a complementary 3D array of nanosize voids is also formed; with patterns, geometry and dimensions depending on the protein ‘building block’ and its mode of ‘packaging’ within the crystal .The composites obtained by ‘filling’ these voids of protein crystals as biotemplates have potential in providing nanostructured, 3D, highly accurate arrays of hybrid materials for applications in fields such as electronic devices  and speThe exploitation of the potential inherent to 3D protein crystal biotemplating calls for the availability of complex infrastructures, including the means, know-how, and the control of protein crystallization and protein crystal stabilization to prevent crystal dissolution; as well as structural analysis by XRD and related advanced spectrophotometric methods in order to monitor the preservation of the parent 3D crystalline structure throughout the templating procedure, and the prevention of distortion resulting from the impact on the template array by the templated material.The feasibility of using such an infrastructure for the monitoring of the stability of a crosslinked lysozyme crystal throughout biotemplating of a crosslinked hydrogel inside its internal voids array was successfully demonstrated in our laboratory . FollowiStabilization of protein crystals by chemical crosslinking is a pre-requisite to controlled 3D biotemplating, as the stability of the crystalline protein template should be preserved throughout the process. This stabilization is mostly affected by glutaraldehyde crosslinking . In vieThe elucidation of the glutaraldehyde crosslinking mechanism and an understanding of its progress throughout the protein crystal structure paved the way for the potential inherent to site-preferred partial crosslinking, enabling the isolation of the homogeneous population of protein products  “liberated” from partially crosslinked crystals by re-dissolution, as demonstrated in our laboratory .A large spectrum of 3D void arrays, exhibiting different shapes and sizes, is available for exploitation from the Protein Data Bank (PDB). Their use is, however, often limited by the commercial availability of an envisaged protein or tedious crystallization procedures and their structural analysis. An alternative route for fishing data out from the PDB might be the use of a “parent” protein building block for the preparation of a series of protein crystals, each exhibiting a different 3D void array. This approach may be materialized by affecting different crystallization conditions: (i) modification of salt and alcohol concentrations, (ii) “directing” the crosslinking of binding proteins by bi-ligand as demonstrated by crosslinking of concanavalin A, yielding ‘diamond-like’ crystals by crosslinking with monosaccharide bi-ligands ; or (iiiAcetivibrio cellulolyticus) as illustrated in To illustrate the feasibility of the latter approach, Wine and coworkers ,83 used An alternative route to affect parent protein–protein molecular interactions affecting crystal “packing” was demonstrated by Cohen-Hadar et al. , using tTakeda et al.  suggeste4, indicating the superiority of smaller crystals for this application.He and colleagues ,87 demonThe feasibility of using bacterial flagella, either extracted from bacterial cells or re-assembled from purified flagellin, was demonstrated by several groups.Salmonella typhimurium . Oxide formation on the surface of a series of biological templates, including flagella mechanically detached from S. typhimurium was also demonstrated by this laboratory [Mao and colleagues  demonstrboratory , using cS. typhimurium flagella reconstructed from isolated and purified flagellin. The hybrid obtained was characterized by HRTEM, X-ray and Raman spectroscopy, followed by electrochemical I-V measurements, indicating that electrical conductivity of the hybrid prepared from reconstructed flagella was superior to the conductivity exhibited by directly extracted flagella.Gopinathan et al.  demonstrProtein-mediated biotemplating has attracted much attention throughout the past decade, culminating in data proving the feasibility of a wide spectrum of biotemplating systems demonstrated in one, two and three dimensions. All these data are supported by advanced, state-of the-art characterization technologies.The preliminary feasibility demonstrations of several applications made available by protein-mediated biotemplating was also demonstrated with an emphasis on nanoelectronics and biosensing. It appears, however, that most protein-mediated biotemplating systems described so far did not culminate in feasibility demonstrations for their potential applications. Therefore, it appears that protein-mediated biotemplating-related research and development (R+D) efforts should address the challenges of the identification and demonstration of new applications, and focus on further developing the already described initiations.Future advancement of such a technology should also aim to solve important practical aspects as bringing results from bench to practice, including the provision of the means of contact or connection of the hybrids thus obtained to working/detecting systems, as well as data on storage and operational stability. The process of coping with future R+D challenges may benefit from further exploitation of the potential inherent to the incorporation of additional protein engineering and chemical modification-based methods for the manipulation and performance improvement of protein templates.


The hypothalamic non-apeptide oxytocin (OT) has several physiological functions, ranging from lactation to social attachment  neurons, showed that OT induces membrane hyperpolarization and consequently inhibits the nociceptive transmission by Ca2+-dependent mechanisms  neurons and ex vivo patch-clamp as well as behavioral analyses, suggest that OT inhibits the nociceptive input either directly , for which OT has a physiologically relevant affinity, cannot be ruled out  abolished the OT-induced Ca2+ influx. These results were confirmed by replicating both the nociceptive DRG neurons isolated from wild-type animals and the modest OT-induced analgesia observed in TRPV1 knock-out mice. Further electrophysiological, planar lipid bilayer, and in silico experiments showed that OT acts as a partial agonist of TRPV1 channels and induces a strong desensitization of these channels, under nociceptive stimulation. This pioneering study strongly suggests that at peripheral level, OT-induced analgesia passes at least in part, through a direct TRPV1 interaction.Nersesyan et al. was first to show that aversive behaviors induced by an intracutaneous (i.c.) injection of capsaicin into an animal hindpaw, were reduced in animals concomitantly treated with i.c. OT. One can therefore propose that the OT effect is a result of OTR activation at the peripheral nerve endings  mediated by GABA. These data suggest that OT recruits GABAergic interneurons, which in turn, activate GABAB receptors and consequently inhibit the capsaicin-sensitive neurons. Interestingly, TRPV1 and GABAB receptors were found to be coexpressed in primary nociceptive neurons. Additionally, the TRPV1 spinal cord expression was enhanced in neuropathic rats, an up-expression corrected by intrathecal OT treatment. Accordingly, the OT-induced analgesia was partially reduced in TRPV1 knockout mice. Like the proposal of Nersesyan et al., this study points out that OT-induced analgesia may rely on both GABA release and modulation of TRPV1 activity.To further explore if OT-induced analgesia in neuropathic pain may occur via TRPV1, Sun et al. first confirmed that intrathecal OT diminishes the aversive behavior in neuropathic animals (Miranda-Cárdenas et al., B receptors, which in turn modulate the TRPV1 function during neuropathic pain. This finding is in line with the observation that GABAB receptors revert the TRPV1 sensitization by noncanonical signaling under pathological pain (Hanack et al., In the past decades, OT has slowly emerged as an important mediator of endogenous analgesia (Poisbeau et al., An intriguing point in Sun et al. study is how exactly, at the spinal level, OT engages the intracellular machinery to modulate the TRPV1 expression. At least two mechanisms can be proposed. The first might be a direct action of OT on TRPV1, as suggested by Nersesyan et al. The second might be that the OT-induced activation of OTR, leads to an intracellular cascade capable of modulating the expression or epigenetic modifications of TRPV1, as suggested by Sun et al. It remains unclear however, how exactly OT modulates the TRPV1 functions and whether it remains true at physiological concentrations. Indeed, while Nersesyan et al. clearly show a direct interaction between OT and TRPV1, neither them nor Sun et al. were able to reveal a strong, behaviorally relevant, OT-mediated TRPV1 modulation. Since OTR was recently involved in functional heterodimers, such as vasopressin or dopamine receptors (Terrillon et al., Finally, although the sources of OT and its spinal effects are neuronal and seem to play a role in endogenous analgesia (Eliava et al., In conclusion, a collection of evidence, briefly explained here, suggests that a triad between OT, GABA, and TRPV1 does exist and sheds a light on new mechanisms involved in OT-induced analgesia, which ultimately is much more complex than initially expected.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.


Rhodovulum sulfidophilum, a marine purple non-sulfur bacterium. Several types of artificial RNAs have been successfully produced in R. sulfidophilum. Purple photosynthetic bacteria produce hydrogen via nitrogenase, and genetic engineering strategies have been investigated to enhance the hydrogen production. This mini review describes the microbial production of these high-value compounds using purple photosynthetic bacteria as the host microorganism.Photosynthetic microorganisms can serve as the ideal hosts for the sustainable production of high-value compounds. Purple photosynthetic bacteria are typical anoxygenic photosynthetic microorganisms and are expected to be one of the suitable microorganisms for industrial production. Purple photosynthetic bacteria are reported to produce polyhydroxyalkanoate (PHA), extracellular nucleic acids and hydrogen gas. We characterized PHA production as a model compound in purple photosynthetic bacteria, especially focused on marine strains. PHA is a family of biopolyesters synthesized by a variety of microorganisms as carbon and energy storage materials. PHA have recently attracted attention as an alternative to conventional petroleum-based plastics. Production of extracellular nucleic acids have been studied in  Cyanobacteria, algae and plants have two photosystems (photosystem I and II), extract electrons from water, and evolve oxygen as a byproduct , extracellular nucleic acids and hydrogen gas as shown in Cupriavidus necator, a hydrogen-oxidizing bacterium, is the most studied bacterium for PHA production and produced about 90% of dry cell weight (wt%) PHA  through three enzyme reactions . KetothiA. vinosum, a purple sulfur bacterium, has Class III type PhaC and extensively studied its biological activity  was produced by a cell free protein expression system and characterized its activity -3-hydroxybutyryl-CoA (3HB-CoA) and did not saturate, suggesting that the PhaCRs was not saturated due to low affinity for the substrate. Generally, PhaC is thought to exist as monomeric and dimeric forms in equilibrium and dimerization of PhaC induced by substrate binding facilitate the PHA polymerization  affect the cell growth and PHA production in R. sulfidophilum  among three types of wavelengths we studied. We found that marine purple non-sulfur bacteria strains hardly accumulated PHA (<5 wt%) under aerobic conditions in the presence of malate and pyruvate. Interestingly, the addition of acetate induced high PHA production (33 wt%) under aerobic conditions. The expression of isocitrate dehydrogenase in the tricarboxylic acid (TCA) cycle decreased under aerobic conditions in the presence of malate and pyruvate and upregulated by the addition of acetate. Considering these results, we proposed that low PHA production under aerobic conditions is caused by low activity of the TCA cycle and its activity was enhanced by the addition of acetate. We found that the expression of PdhR, which is a transcriptional repressor of the pyruvate dehydrogenase complex, was upregulated upon the addition of acetate. The changes in the metabolic state might be induced by the addition of acetate under aerobic conditions and PdhR is involved in this regulation.PHA production was examined in marine purple non-sulfur bacteria under various growth light and oxygen conditions  [P(3HB-co-3HV)], has a lower melting temperature and higher biodegradability compared to P(3HB) depending on the polymer composition  and 3-hydroxyvalerate (3HV). PHA composition affects the mechanical and thermal properties of PHA. Poly(3-hydroxybutyrate) [P(3HB)], homopolymer of 3HB, is a highly crystalline and brittle material. Melting temperature of P(3HB) is around 180°C Rehm, . The copE. coli that do not have PHA degradation pathway  have been found in natural conditions such as freshwater, seawater, and soil and it is reported that some bacteria produced nucleic acids extracellularly  are considered to be involved in the production of extracellular nucleic acids in in vitro transcription . Purple non-sulfur bacteria is known to produce hydrogen via nitrogenase    (Franchi et al., A variety of kinds of large-scale photobioreactor has been investigated for industrial photohydrogen production in purple non-sulfur bacteria. Reactor design, culture light condition and nutrient sources have been investigated and are reviewed elsewhere (Eroglu and Melis, 2, N2 and sunlight energy can be used as a culture medium, biological sources and energies. Genetic tools such as synthetic promoter and transformation method have not been fully established yet in marine purple photosynthetic bacteria, even though we recently developed a transformation method using chemical competent cells of marine purple photosynthetic bacteria (Higuchi-Takeuchi et al., Marine purple photosynthetic bacteria are environmentally friendly microorganisms and can produce high-valuable compounds such as PHA, extracellular nucleic acids, and hydrogen gas. Previously, we demonstrated that marine purple photosynthetic bacteria were able to produce PHA even in seawater (Higuchi-Takeuchi et al., MH-T and KN conceptualized and wrote the manuscript.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.


Electronic health  tools are becoming increasingly popular for helping patients’ self-manage chronic conditions. Little research, however, has examined the effect of patients using eHealth tools to self-report their medication management and use. Similarly, there is little evidence showing how eHealth tools might prompt patients and health care providers to make appropriate changes to medication use.The objective of this systematic review was to determine the impact of patients’ use of eHealth tools on self-reporting adverse effects and symptoms that promote changes to medication use. Related secondary outcomes were also evaluated.MEDLINE, EMBASE, and CINAHL were searched from January 1, 2000, to April 25, 2018. Reference lists of relevant systematic reviews and included articles from the literature search were also screened to identify relevant studies. Title, abstract, and full-text review as well as data extraction and risk of bias assessment were performed independently by 2 reviewers. Due to high heterogeneity, results were not meta-analyzed and instead presented as a narrative synthesis.A total of 14 studies, including 13 randomized controlled trials (RCTs) and 1 open-label intervention, were included, from which 11 unique eHealth tools were identified. In addition, 14 RCTs found statistically significant increases in positive medication changes as a result of using eHealth tools, as did the single open-label study. Moreover, 8 RCTs found improvement in patient symptoms following eHealth tool use, especially in adolescent asthma patients. Furthermore, 3 RCTs showed that eHealth tools might improve patient self-efficacy and self-management of chronic disease. Little or no evidence was found to support the effectiveness of eHealth tools at improving medication recommendations and reconciliation by clinicians, medication-use behavior, health service utilization, adverse effects, quality of life, or patient satisfaction. eHealth tools with multifaceted functionalities and those allowing direct patient-provider communication may be more effective at improving patient self-management and self-efficacy.Evidence suggests that the use of eHealth tools may improve patient symptoms and lead to medication changes. Patients generally found eHealth tools useful in improving communication with health care providers. Moreover, health-related outcomes among frequent eHealth tool users improved in comparison with individuals who did not use eHealth tools frequently. Implementation issues such as poor patient engagement and poor clinician workflow integration were identified. More high-quality research is needed to explore how eHealth tools can be used to effectively manage use of medications to improve medication management and patient outcomes. Use of the internet has increased considerably since the early 1990s. The World Bank reports that almost 44% of people across the globe used the internet in 2015, compared with 0.25% in 1993 , while aWell-functioning eHealth tools can help patients better understand their health  and may The ability of patients to use eHealth tools to better manage medication by reporting feedback on symptoms and use of medications directly to health care providers has not been comprehensively explored in the literature. Similarly, there is little evidence showing how eHealth tools might provide prompts to patients and health care providers to make appropriate changes to medication use based on this feedback. A synthesis of this literature will provide greater understanding of what eHealth tool design features may be helpful in patient reporting of medication-related experiences and outcomes.The objective of this systematic review was to determine the impact of patients’ use of eHealth tools on self-reporting adverse effects and symptoms that promote changes to medication use. The PICO model was used to focus the objective of the review, as seen in This systematic review was performed following steps outlined by Cochrane’s Effective Practice and Organization of Care group and reported based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement . A totalFor the purposes of this review, an eHealth tool was considered to be any internet-based intervention, including mobile health apps, used by patients for clinical purposes that focused on improving patient health and clinical outcomes. The term PHR refers to an eHealth tool wherein a patient has access to and can enter or edit their own health data. The population investigated was community-dwelling individuals of any age in an outpatient setting.For a study to be included, the eHealth tool must have allowed patients (or caregivers) to enter information directly ; included self-reporting functionalities focusing on medication monitoring, contain a medication monitoring or use component, or specifically incorporating the option for the patient or caregiver to enter symptoms including adverse effects; and needed to focus specifically on medication use, clinical outcomes, or symptom reporting following use of the eHealth tool. Any eHealth tools involving changes in medication reconciliation and recommendations made to changes in drug therapy were also included.Exclusion criteria were conference abstracts; qualitative studies; articles without a comparator group; articles that did not report on at least one medication-related outcome; articles where self-management strategies focused on lifestyle modification, behavioral interventions, or nondrug interventions; articles focused solely on the validation of an eHealth tool; articles focused on methodological or technical aspects of eHealth interventions; articles containing nonempirical information; articles that synthesized information about multiple eHealth tools in an article ; and eHealth tools used by regulatory agencies to report adverse drug events (ADEs).All potentially relevant articles were uploaded into DistillerSR software, which was used throughout the selection process. Potentially relevant articles underwent title, abstract, and full-text review. Articles that met inclusion criteria proceeded to data abstraction and risk of bias assessment. Articles not meeting inclusion criteria were excluded at both levels. Title and abstract review were performed independently by 2 reviewers from a pool of 5 reviewers. Of these, 1 reviewer went through the reference lists of all the articles included in this study. Another reviewer went through reference lists of relevant systematic reviews identified during the literature search. Potentially relevant articles were identified. These articles went through abstract review by 2 reviewers. Studies found not to fit inclusion criteria after abstract review were excluded. Full-text review was performed independently by 5 reviewers. The kappa scores were calculated to determine agreement among reviewers who conducted review of titles and abstracts. All kappa scores calculated were greater than .93. Conflicts were resolved by consensus.Data extraction and risk of bias assessment were performed for each study independently by 2 reviewers. Data extracted included study design and setting, participant demographics, number of participants in each group, intervention components, comparator group components, eHealth tool functionality measured, and results and significance levels for each outcome measure. Conflicts in data extraction were resolved by consensus. Risk of bias assessment used questions recommended by the Agency for Healthcare Research and Quality’s 2014 publication Methods Guide for Effectiveness and Comparative Effectiveness Reviews  and was The primary and secondary outcomes are listed in Subgroup analyses were performed to investigate differences in treatment effect present because of (1) age of participants, (2) patients with specific conditions targeted by intervention, and (3) different features and functionalities of the included eHealth tools.A total of 3515 articles were generated from database and reference searching, resulting in 2489 potential articles that were screened based on their titles and abstracts, after duplicates were removed. Furthermore, 75 full-text articles were assessed for eligibility, of which 14 were included in this systematic review  -31 and 1Of the 13 RCTs included in this review, 2 studies were cluster RCTs ,26. The A total of 4 RCTs focused on pediatric and adolescent asthma patients ,23-25; 1double-dummy style intervention, where both groups used Web-based PHRs to record information that differed only in content. Cho et al [In 3 studies, use of an eHealth tool in the intervention group was compared with usual care plus links to relevant websites ,25,28. Sho et al  comparedho et al  utilizedho et al  used a sho et al  developeho et al  used a mho et al . Carlsenho et al  used an From the 14 included studies, 11 unique eHealth tools were described. The 2 RCTs by Joseph et al ,25 utiliAll 11 eHealth tools from all 14 studies included a component where patients could self-report medication management information or changes, including symptoms, health data, adverse effects, or ADEs. A total of 12 studies included Web-based patient questionnaires or surveys ,21,23-32The results of each study by outcome can be seen in A total of 6 RCTs -22,24,27P=.035). A nonsignificant trend approaching significance was seen for decreased number of over-the-counter medications used in the eHealth tool group (P=.05) [P<.001). They also found that a significantly higher proportion of patients in the intervention group had medications initiated or dosages changed for hypertension  and hyperlipidemia .Chrischilles et al  found a  (P=.05) . Grant e (P=.05)  found a P=.01). Simon et al [21=10.5, P=.001). Carlsen et al [P<.001) [Joseph et al  found thon et al  found then et al  showed ten et al  showed t[P<.001) .P=.36). Fiks et al [In contrast, Cho et al  found noks et al  provided1c) [A total of 9 RCTs ,27,30,311c) , and 2 fP=.001) and that the active group had less frequent flare-ups . Gustafson et al [P=.01).Fiks et al  reportedon et al  found anP=.009), symptom days , days of restricted activity , and school days missed . In another study, Joseph et al [P=.01). Following subgroup analysis, it was found that teenagers with moderate to severe asthma had fewer symptom days , total school days missed , school days missed because of asthma , and days of restricted activity .Joseph et al  found thph et al  reportedP=.04). Ahmed et al [1c ; however, they did find a statistically significant decrease in waist circumference between intervention and control . Cho et al [1c in the active group  at 30 months. Grant et al [1c and for percentage of patients at target HbA1c levels. Similarly, Carlsen et al [P=.18).Simon et al  reporteded et al  reporteded et al  found noho et al  did findnt et al  found noen et al  found noA total of 5 RCTs measured this outcome ,23,26,30P=.01). They also found a positive significant effect of intervention on ACQ score when mediated by information competence . Schnipper et al [P<.001). Ahmed et al [Gustafson et al  found ther et al  found thed et al  found thed et al  found thed et al  found noA total of 3 RCTs measured this outcome, all using measures of medication adherence as a surrogate for medication use behavior ,23,24. NP=.01). In addition, Schnipper et al [P=.04). The number of medication discrepancies per patient with the potential for harm approached significance as a result of using eHealth tools (P=.05) [A total of 3 RCTs reported on this outcome ,20,26. OCho et al  acknowleP value: .02 to <.001) relative to usual care. Descriptive evidence from Fiks et al [A total of 1 open-label intervention  and 5 RCks et al  showed 1ks et al  providedks et al  or ADEs ks et al ,28 foundP=.01). The remaining 5 studies [A total of 6 RCTs reported health service utilization outcomes ,27,28,30 studies ,27,28,30A total of 4 studies measured this outcome ,30-32, a21=8.38, P=.004). Fiks et al [A total of 2 RCTs ,27 and 1ks et al  providedA total of 4 RCTs investigated the use of eHealth tools in children and teens with asthma ,23-25. TSubgroup analysis also found that multifaceted interventions combining use of eHealth tools with clinician support or case management and eHealth tools utilizing direct patient-provider communication might be more effective at improving some aspects of patient self-management and self-efficacy ,26,30,31Many studies reported barriers to the implementation of eHealth tools. The most common barrier was lack of participant engagement, resulting in low eHealth tool utilization rates. This was reported by 9 of the 14 studies -27,31,32Evidence from 4 RCTs and 1 open-label intervention ,24,27,32Evidence was found that eHealth tools improved the outcome of patient self-management and self-efficacy. Subgroup analysis found that eHealth tools that allow patients and clinicians to communicate directly, and multifaceted interventions combining eHealth tools with clinician support and case management might lead to greater increases in patient self-management and self-efficacy. It is notable that more significant improvements were found for more objective outcome measures, such as number of medication changes and clinical signs, and less were found for more subjective outcome measures such as self-management and self-efficacy. It may also be that sample sizes were too small to detect differences, particularly if this was not the primary objective for these studies. It is likely that the eHealth tools under investigation either did not provide effective content or functionalities to help participants improve self-management and medication management in participants or the tools used to measure these outcomes were not able to detect any differences between groups. Another possibility is the lack of patient understanding of chronic disease and poor perception of health goals.It was thought that eHealth tools that focus on improvement of patient self-efficacy and self-management might lead to improved medication-use behavior, which in turn may lead to changes in medication use, identification of real or potential ADEs, improvement in signs and symptoms, and overall improvement in HRQoL. However, there is not enough evidence to draw conclusions as to the effectiveness of eHealth tools for identification of adverse effects, improving medication-use behavior, increasing recommendations to medication therapies and improving medication reconciliation, improving health service utilization, and improving overall health status and patient satisfaction. Only a small number of included studies investigated these outcomes; it is likely that with such a small overall sample size, it was not possible to find differences between groups.As with most systematic reviews on the subject of eHealth tools ,33,54-57A 2012 systematic review by Ammenwerth et al found that use of patient portals linked to a PHR led to significant increases in medication adjustments in diabetic patients . Other rOverall, this review supports findings by Ammenwerth et al  that intTo our knowledge, this is the first systematic review of eHealth interventions focusing on patient self-reporting of symptoms and adverse effects. The review’s search strategy was augmented by reference searching. This review was limited to studies that included medication-related outcomes. The majority of studies included in this review were RCTs, most of which were of moderate quality.eHealth and believe future research should focus on establishing better consensus for this term. There was considerable variety among the interventions in the studies, some of which included features such as direct health care provider follow-up, thus making it more complicated to determine which outcomes could be specifically attributable to using an eHealth tool. As this review also examined different populations of varying sample sizes and medical conditions for eHealth tools, it may be difficult to detect differences and generalize findings and conclusions. We did not include qualitative studies in our review because the goal of our study was to better understand the effectiveness and impact of changes to medication regimens based on quantifiable differences in using eHealth tools for self-reporting adverse effects and symptoms that promote changes to medication use versus a comparator. We value the insight of qualitative studies that have been investigated elsewhere [This review also has a number of weaknesses. It was limited to studies published in English, which may have excluded relevant articles. No searching of grey literature was performed, as we focused on empirical work published in academic journals, and so it is possible that some early non–peer-reviewed reports may have been missed. We acknowledge the lack of definitional clarity surrounding the term lsewhere ,60. AddiWhere possible, health care providers should encourage patient use of eHealth tools for symptom and adverse effect self-reporting. eHealth tools may be especially useful for reducing symptoms in certain populations, for example, children and teenagers with asthma. eHealth tools might also encourage patients to improve self-management behaviors and participate in shared decision making with clinicians. Having information from the EMR entered directly into the eHealth tool may reduce the burden on the patient to routinely update their clinical information (something that only highly motivated patients are likely to do regularly) . CliniciThere is a paucity of primary research articles investigating eHealth tools and their impact on medication use. Studies are generally small and of moderate quality. Large-scale RCTs focusing on the use of eHealth tools for medication and symptom management should be undertaken to establish more high-quality evidence. This is especially important given how ubiquitous the use of medication is. Furthermore, the effects of patient self-management and self-efficacy on medication use and symptom experience are not well studied; more research in this area could help drive creation of medication-focused eHealth tools. Low patient engagement and eHealth tool utilization were commonly noted implementation barriers; it could be that patients were not engaged in eHealth tool use enough for them to feel an impact on their satisfaction with health care or overall quality of life. Descriptive evidence shows low proportions of patients felt that eHealth tools improved their care or communication with providers, indicating that development of eHealth tools should focus on functionalities and outcomes that are important to the patient. This may be achieved by utilizing research on patient motivation and behavior change to increase patient engagement ,24,25.The results of this review show initial and promising findings that specialized eHealth tools can be used for reporting and monitoring of symptoms and medication-related adverse effects and some evidence that use of eHealth tools have the potential to identify instances where changes in medication use may be appropriate. A modest amount of mixed evidence was found, demonstrating that eHealth tools can improve patient self-management and self-efficacy. Very little or no evidence was found to demonstrate that use of eHealth tools could increase numbers of medication recommendations or improve medication-taking behavior, health services utilization, identification of adverse effects, overall health status, and patient satisfaction. eHealth tools may be more effective at promoting medication changes and improving patient self-management and self-efficacy if they provide mechanisms for direct patient-provider communication and may be more effective in certain populations such as children and teenagers with asthma.


Recently Christian Hudert and colleagues from the Charité in Berlin published a study about genetic determinants in pediatric non-alcoholic liver disease . Non-alDue to the current increase in the incidence of liver diseases a better understanding of their pathophysiology is of major importance (Jansen et al., 2017; Vartak


Sci., 2019, DOI: ; 10.1039/c8sc05721d.Correction for ‘Facile N-functionalization and strong magnetic communication in a diuranium( The authors regret that The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.


Alzheimer's disease (AD), the most common type of dementia among the elderly, is caused by progressive neural death that results in impaired memory, thinking skills and, eventually, the ability to carry out simple tasks. Unfortunately, approximately 5% of people over the age of 65 suffer from AD, and the prevalence of AD increases with aging (Udeochu et al., Investigators have proposed that the onset and progression of AD is contributed by pathogenic microbes although definitive evidence has not been presented (Sjogren et al., Based on unbiased approaches and large-scale data sets from several brain banks and cohort studies, this is the first study to provide strong evidence supporting the controversial hypothesis that viruses play an essential role in regulatory genetic networks that are believed to lead to AD. Identifying links to viruses may help scientists interested in developing potential new treatment strategies.Several important questions are raised from this phenomenal study. First, does herpes virus cause the onset or progression of AD? Eimer and colleagues recently reported that Aβ traps herpes viruses in insoluble amyloid, and active herpes infections in brain accelerate amyloid deposition, indicating that herpes infection may promote AD pathology directly via amyloid-mediated pathological pathways (Eimer et al., Second, whether herpesviruses regulate, or are regulated by AD-associated genes? Will anti-herpes drugs be effective against early onset of AD? This study established a strong connection between multiple viruses, especially HHV-6A, and AD risk genes, including PSEN1, BACE1, and APBB2 which are implicated in regulation of Aβ production. Besides, several recent studies show that Aβ is an antimicrobial protein of the body's innate immune system, capable of providing immediate, effective protection from infection with pathogens like herpes viruses in both cultured human brain cells and animal models of AD (Kumar et al., Third, is miR-155 a regulator of anti- or pro-viral activity in early AD? Could miR-155 gain-of-function help with Aβ clearance and neuroprotection in AD? MiR-155 has been reported as an important regulator of T cell and microglia in response to neurodegeneration (Song and Lee, Lastly, there remain some open issues that future studies will need to address about the Readhead et al. findings. For example, HHV-6A/B and HHV-7 are considered lymphotropic rather than neurotrophic (Berneman et al., X-WS drafted an initial version of this commentary. C-ML and Z-QT revised and finalized the text. All authors 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.