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The present review summarizes the current knowledge concerning physiological effects of resveratrol (RSV) with emphasis on the RSV action on female reproductive processes. The review outlines provenance, properties, mechanisms of action, physiological and therapeutic actions of RSV on female reproduction and other physiological processes, as well as areas of possible application of R. This review is based on the search for the related full papers indexed in Medline/Pubmed, Web of Science and SCOPUS databases between the year 2000 and 2021 according to the criteria of preferred reporting items for systematic reviews and meta-analyses extension for scoping reviews and other related guidelines. The analysis of the available information suggests that RSV has a number of properties which enable its influence on various physiological processes including female reproduction at various regulatory levels via various extra- and intracellular signalling pathways. Despite some contradictions and limitations in the available data, they indicate applicability of both stimulatory and inhibitory effects of RSV for control and influence of various reproductive and non-reproductive processes and treatment of their disorders in phytotherapy, animal production, medicine, biotechnology and assisted reproduction. To establish the clinical efficacy of RSV, further high quality studies are needed.

The cognitive deficits of schizophrenia are linked to imbalanced excitatory and inhibitory signalling in the prefrontal cortex (PFC), disrupting gamma oscillations. We previously demonstrated that two mGlu5 receptor-positive allosteric modulators (PAMs), VU0409551 and VU0360172, restore cognitive deficits in the sub-chronic PCP (scPCP) rodent model for schizophrenia via distinct changes in PFC intracellular signalling molecules. Here, we have assessed ex vivo gamma oscillatory activity in PFC slices from scPCP rats and investigated the effects of VU0409551 and VU0360172 upon oscillatory power. mGlu5 receptor, protein kinase C (PKC), and phospholipase C (PLC) inhibition were also used to examine ‘modulation bias’ in PAM activity. The amplitude and area power of gamma oscillations were significantly diminished in the scPCP model. Slice incubation with either VU0409551 or VU0360172 rescued scPCP-induced oscillatory deficits in a concentration-dependent manner. MTEP blocked the PAM-induced restoration of oscillatory power, confirming the requirement of mGlu5 receptor modulation. Whilst PLC inhibition prevented the power increase mediated by both PAMs, PKC inhibition diminished the effects of VU0360172 but not VU0409551. This aligns with previous reports that VU0409551 exhibits preferential activation of the phosphatidylinositol-3-kinase (PI3K) signalling pathway over the PKC cascade. Restoration of the excitatory/inhibitory signalling balance and gamma oscillations may therefore underlie the mGluR5 PAM-mediated correction of scPCP-induced cognitive deficits.

The cannabinoid type 2 receptor (CB2R) is an important therapeutic target for pain and inflammatory disorders. G protein-coupled receptors (GPCRs) are conventionally thought to signal exclusively at the plasma membrane; however, recently this has been challenged by the notion of intracellular signalling receptors. Better understanding of GPCR location requires tools that can differentiate cell surface versus subcellular receptors as well as accessing different parts of the body. Herein, we report the synthesis and pharmacological evaluation of polar chromenopyrazole-based CB2R-selective agonists that contain short peptides that could be useful tools for interrogating CB2R.

Biological therapies have improved the outcomes of several major inflammatory, autoimmune and also neoplastic disorders. Those directed towards cytokines or other soluble mediators, cell-surface molecules or receptors or various components of intracellular signalling pathways may be associated with the occurrence of infections whose diversity depends on the particular immune target. In this context and following a keynote lecture given by one of us at the European League Against Rheumatism meeting on June 2018, a multidisciplinary group of experts deeply involved in the use of targeted and biological therapies in rheumatoid and psoriatic arthritis decided to summarise their recent vision of the immunological basis and epidemiology of infections occurring during targeted and biological therapies, and provide useful indications for their management and prevention.

Plants produce a number of biological active substances with healthy benefits. Resveratrol (3,5,4′-trihydroxystilbene), a polyphenol produced by plants has been associated with many health beneficial properties, including its ability to induce mitochondrial biogenesis and fight against health problems such as obesity, inflammation, heart diseases, cancers among others. Mitochondrial dysfunction is recognized as central to the pathogenesis and development of many diseases. Thus the present review describes how resveratrol (RSV) may counteract physiological and age-related diseases/disorders through its effect on mitochondrial biogenesis and function. In addition, we discuss the chemistry, main sources, and the doses of RSV shown in previous studies to be efficient for the prevention and treatment of different diseases. Through its ability to improve mitochondrial dysfunction, RSV can be used in the prevention and/or treatment of human physiological diseases. However, more research for optimal dose in a human scale is still relevant. This review brings new hope to the therapy of physiological diseases as it will provide useful future perspectives for the planning of clinical studies on RSV and mitochondrial dysfunction-related diseases.

The CaMK subfamily of Ser/Thr kinases are regulated by calmodulin interactions with their C-terminal regions. They are exemplified by Ca2+/calmodulin dependent protein kinase 1δ which is known as CaMK1D, CaMKIδ or CKLiK. CaMK1D mediates intracellular signalling downstream of Ca2+ influx and thereby exhibits amplifications of Ca2+signals and polymorphisms that have been implicated in breast cancer and diabetes. Here we report the backbone 1H, 13C, 15N assignments of the 38 kDa human CaMK1D protein in its free state, including both the canonical bi-lobed kinase fold as well as the autoinhibitory and calmodulin binding domains.

In recent years, several novel aspects of GPCR pharmacology have been described, which are thought to play a role in determining the in vivo efficacy of a compound. Fluorescent ligands have been used to study many of these, which have also required the development of new experimental approaches. Fluorescent ligands offer the potential to use the same fluorescent probe to perform a broad range of experiments, from single‐molecule microscopy to in vivo BRET. This review provides an overview of the in vitro use of fluorescent ligands in further understanding emerging pharmacological paradigms within the GPCR field, including ligand‐binding kinetics, allosterism and intracellular signalling, along with the use of fluorescent ligands to study physiologically relevant therapeutic agents.

Colorectal cancer (CRC) is one of the most prevalent cancers, driven by several factors including deregulations in intracellular signalling pathways. Small extracellular vesicles (sEVs) are nanosized protein-packaged particles released from cells, which are present in liquid biopsies. Here, we characterised the proteome landscape of sEVs and their cells of origin in three CRC cell lines HCT116, HT29 and SW620 to explore molecular traits that could be exploited as cancer biomarker candidates and how intracellular signalling can be assessed by sEV analysis instead of directly obtaining the cell of origin itself. Our findings revealed that sEV cargo clearly reflects its cell of origin with proteins of the PI3K-AKT pathway highly represented in sEVs. Proteins known to be involved in CRC were detected in both cells and sEVs including KRAS, ARAF, mTOR, PDPK1 and MAPK1, while TGFB1 and TGFBR2, known to be key players in epithelial cancer carcinogenesis, were found to be enriched in sEVs. Furthermore, the phosphopeptide-enriched profiling of cell lysates demonstrated a distinct pattern between cell lines and highlighted potential phosphoproteomic targets to be investigated in sEVs. The total proteomic and phosphoproteomics profiles described in the current work can serve as a source to identify candidates for cancer biomarkers that can potentially be assessed from liquid biopsies.

Background and Purpose: The intracellular signalling molecule cGMP, formed by NO-sensitive GC (NO–GC), has an established function in the vascular system. Despite numerous reports about NO-induced cGMP effects in the heart, the underlying cGMP signals are poorly characterized. Experimental Approach: Therefore, we analysed cGMP signals in cardiac myocytes and fibroblasts isolated from knock-in mice expressing a FRET-based cGMP indicator. Key Results: Whereas in cardiac myocytes, none of the known NO–GC-activating substances (NO, GC activators, and GC stimulators) increased cGMP even in the presence of PDE inhibitors, they induced substantial cGMP increases in cardiac fibroblasts. As cardiac myocytes and fibroblasts are electrically connected via gap junctions, we asked whether cGMP can take the same route. Indeed, in cardiomyocytes co-cultured on cardiac fibroblasts, NO-induced cGMP signals were detectable, and two groups of unrelated gap junction inhibitors abolished these signals. Conclusion and Implication: We conclude that NO-induced cGMP formed in cardiac fibroblasts enters cardiac myocytes via gap junctions thereby turning cGMP into an intercellular signalling molecule. The findings shed new light on NO/cGMP signalling in the heart and will potentially broaden therapeutic opportunities for cardiac disease.

When produced transiently and moderately, reactive oxygen species (ROS) are not just by-products of substrate oxidation, they are also chemicals involved in intracellular signalling. The objective of this review is to explore the intracellular signaling aspects of reactive oxygen species in influencing feeding behaviour. Substrates such as glucose and lipids promote the generation of reactive oxygen species, primarily through mitochondria and, to a lesser extent, through NADPH oxidases. Under the influence of substrate level, the level of ROS generated in hypothalamic neurons such as NPY/AgRP and POMC neurons directly influences their activity and, as a result, affects downstream neurons located in other parts of the hypothalamus such as the ventromedial nucleus (VMN), the paraventricular nucleus (PVN), and the lateral hypothalamus. The activation of the POMC neuronal population is caused by an increase in ROS levels, whereas the activation of the NPY/AgRP neurons is caused by a decrease in ROS levels. The activation of these neurons determines feeding behaviour, which can be satiety if POMC neurons are activated or increased food intake if NPY/AgRP neurons are activated.

G protein coupled receptors (GPCRs) translate the actions of hormones into intracellular signalling events. Mutations in GPCRs can prevent their correct expression and trafficking to the cell surface and cause disease. We use single cell measurements in HEK293 cells to show that the balance between endoplasmic reticulum (ER) and cell surface localisation of the Vasopressin 2 receptor (V2R) varies significantly from cell to cell. We find that mutations in the V2R affect the proportion of cells able to send this GPCR to the cell surface but do not prevent all cells in the population from correctly trafficking the mutant receptors. These findings reveal that the ability of cells to correctly traffic V2R to the cell surface depends not only on the expressed V2R mutant but also on the individual cell environment.Significance statementMissense mutations in the Vasopressin 2 Receptor (V2R) cause Nephrogenic Diabetes Insipidus. Some of these mutations prevent correct expression and trafficking of V2R to the cell surface resulting in a loss-of-function. We show -using single cell measurements-that the balance between endoplasmic reticulum and cell surface localisation of the V2R varies significantly from cell to cell, independent from its expression level. Mutations affect the proportion of cells able to send V2R to the cell surface but do not prevent all cells in the population from correctly trafficking the mutant receptors. Hence, the ability of cells to correctly traffic V2R to the cell surface depends not only on expressed V2R mutant but also on the cell environment.

Objectives Receptor Activity-Modifying Protein 2 (RAMP2) is a chaperone protein which allosterically binds to and interacts with the glucagon receptor (GCGR). The aims of this study were to investigate the effects of RAMP2 on GCGR trafficking and signalling in the liver, where glucagon (GCG) is important for carbohydrate and lipid metabolism. Methods Subcellular localisation of GCGR in the presence and absence of RAMP2 was investigated using confocal microscopy, trafficking and radioligand binding assays in human embryonic kidney (HEK293T) and human hepatoma (Huh7) cells. Mouse embryonic fibroblasts (MEFs) lacking the Wiskott-Aldrich Syndrome protein and scar homologue (WASH) complex and the trafficking inhibitor monensin were used to investigate the effect of halted recycling of internalised proteins on GCGR subcellular localisation and signalling in the absence of RAMP2. NanoBiT complementation and cyclic AMP assays were used to study the functional effect of RAMP2 on the recruitment and activation of GCGR signalling mediators. Response to hepatic RAMP2 upregulation in lean and obese adult mice using a bespoke adeno-associated viral vector was also studied. Results GCGR is predominantly localised at the plasma membrane in the absence of RAMP2 and exhibits remarkably slow internalisation in response to agonist stimulation. Rapid intracellular accumulation of GCG-stimulated GCGR in cells lacking the WASH complex or in the presence of monensin indicates that activated GCGR undergoes continuous cycles of internalisation and recycling, despite apparent GCGR plasma membrane localisation up to 40 min post-stimulation. Co-expression of RAMP2 induces GCGR internalisation both basally and in response to agonist stimulation. The intracellular retention of GCGR in the presence of RAMP2 confers a bias away from β-arrestin-2 recruitment coupled with increased activation of Gαs proteins at endosomes. This is associated with increased short-term efficacy for glucagon-stimulated cAMP production, although long-term signalling is dampened by increased receptor lysosomal targeting for degradation. Despite these signalling effects, only a minor disturbance of carbohydrate metabolism was observed in mice with upregulated hepatic RAMP2. Conclusions By retaining GCGR intracellularly, RAMP2 alters the spatiotemporal pattern of GCGR signalling. Further exploration of the effects of RAMP2 on GCGR in vivo is warranted., Graphical abstract Image 1, Highlights • The glucagon receptor rapidly recycles to the cell membrane in the absence of RAMP2. • RAMP2 promotes internalisation of the glucagon receptor basally and after agonist stimulation. • RAMP2 confers signalling bias to the glucagon receptor with a reduction in β-arrestin-2 recruitment. • A novel NanoBiT bystander assay demonstrates increased endosomal activation of Gαs with RAMP2. • RAMP2 has time-dependent effects on glucagon-stimulated cAMP accumulation.

Developing a model for a signalling pathway requires several iterations of experimentation and model refinement to obtain an accurate model. However, the implementation of such an approach to model a signalling pathway induced by a poorly-known stimulus can become labour intensive because only limited information on the pathway is available beforehand to formulate an initial model. Therefore, a large number of iterations are required since the initial model is likely to be erroneous. In this work, a numerical scheme is proposed to construct a time-varying model for a signalling pathway induced by a poorly-known stimulus when its nominal model is available in the literature. Here, the nominal model refers to one that describes the signalling dynamics under a well-characterised stimulus. First, global sensitivity analysis is implemented on the nominal model to identify the most important parameters, which are assumed to be piecewise constants. Second, measurement data are clustered to determine temporal subdomains where the parameters take different values. Finally, a least-squares problem is solved to estimate the parameter values in each temporal subdomain. The effectiveness of this approach is illustrated by developing a time-varying model for NF-[inline-formula removed]B signalling dynamics induced by lipopolysaccharide in the presence of brefeldin A.

Membrane contact sites (MCSs) are specialized subcellular compartments formed by closely apposed membranes from two organelles. The intermembrane gap is separated by a distance ranging from 10 nm to 35 nm. MCSs are typically maintained through dynamic protein-protein and protein-lipid interactions. These inter-membrane contact sites constitute important intracellular signaling hotspots to mediate a plethora of cellular processes, including calcium homeostasis, lipid metabolism, membrane biogenesis and organelle remodeling. In recent years, a series of genetically-encoded probes and chemogenetic or optogenetic actuators have been invented to aid the visualization and interrogation of MCSs in both fixed and living cells. These molecular tools have greatly accelerated the pace of mechanistic dissection of membrane contact sites at the molecular level. In this review, we present an overview on the latest progress in this endeavor, and provide a general guide to the selection of methods and molecular tools for probing interorganellar membrane contact sites.

The current existing therapies for severe cases of systemic lupus erythematosus (SLE) patients are still limited. Intravenous immunoglobulin (IVIGs), which are purified from the plasma of thousands of healthy human donors, have been profiled as efficacious and life-saving options for SLE patients refractory to conventional therapy. The specific mechanism of action by which IVIGs generate immunomodulation in SLE is not currently understood. In this manuscript, we reviewed some of the hypothesis that have been postulated to explain the IVIG effects, including those on T and B cell intracellular signalling and activation, as well as the interferon signalling pathways involved in the detection of nucleic acids and the defective removal of immune complexes and debris.

The metabolic charts memorized in early biochemistry courses, and then later forgotten, have come back to haunt many immunologists with new recognition of the importance of these pathways. Metabolites and the activity of metabolic pathways drive energy production, macromolecule synthesis, intracellular signalling, post-translational modifications and cell survival. Immunologists who identify a metabolic phenotype in their system are often left wondering where to begin and what does it mean? Here, we provide a framework for navigating and selecting the appropriate biochemical techniques to explore immunometabolism. We offer recommendations for initial approaches to develop and test metabolic hypotheses and how to avoid common mistakes. We then discuss how to take things to the next level with metabolomic approaches, such as isotope tracing and genetic approaches. By proposing strategies and evaluating the strengths and weaknesses of different methodologies, we aim to provide insight, note important considerations and discuss ways to avoid common misconceptions. Furthermore, we highlight recent studies demonstrating the power of these metabolic approaches to uncover the role of metabolism in immunology. By following the framework in this Review, neophytes and seasoned investigators alike can venture into the emerging realm of cellular metabolism and immunity with confidence and rigour. This Review from Jeff Rathmell and colleagues serves as a guide to immunologists on how to select the appropriate tools and techniques to interrogate metabolism in their experimental systems. The authors provide advice for avoiding common mistakes and on how best to employ metabolomics.

In this issue of Systems Biology and Physiology Reports A.A. Martyanov and M.A. Panteleev suggested a review on platelet intracellular signalling network, which is a second part in the discussion on the molecular relationships between platelet activation and responses [1]. The review contains seven thousands words and two hundred references and yet it is not complete, as there are still unclear parts in platelet signalling, especially in its inhibition [2-4]. In an effort to comprehend the platelet activation pattern, I drew a signalling scheme based on the review and data from other authors [2, 3].

Mechanical stretch activates several intracellular signalling networks and modulates gene expressions which initiate structural and functional remodelling in cardiomyocytes. Various durations, loads and frequencies of mechanical forces induce differing response mechanisms. Cardiomyocytes remodel in response to mechanical stress both during cardiac development as well as in disease conditions. In both of these situations, the cytoskeleton has a key role in sensing mechanical stress and in mediating adaptive or maladaptive changes within the cardiomyocyte. Cytoskeleton which is highly sensitive and adaptive to mechanical forces acts as a signal integrator for mechanical and structural inputs. The information is transmitted throughout the cardiomyocyte to ensure that the cell respond and adapt appropriately. Mechanical stretch induces secretion or synthesis of molecules with autocrine and paracrine effects. Energy metabolism is also altered in adult cardiomyocytes, which hypertrophy in response to mechanical stress.

Resistance training is performed by a wide range of populations, to achieve a diverse range of outcomes in skeletal muscle. Predominantly, such outcomes include the development and/or maintenance of skeletal muscle mass and strength but may also include muscular power or endurance. Blood flow restricted (BFR) resistance exercise is recognized as a novel strategy to achieve such outcomes in skeletal muscle, with much lower-loads (20-50% of one-repetition maximum) than those traditionally employed. The capacity to reduce external loading has made BFR exercise an attractive training modality for individuals who may not tolerate high levels of mechanical stress, such as frail older persons or clinical populations. Moreover, temporarily limiting training loads and/or volume has also generated interest in BFR exercise in athletic cohorts. Despite the promise held by this modality of exercise, there is large heterogeneity in the methods (e.g., BFR methodology and exercise prescription) adopted by practitioners, which often leads to variable training outcomes. Consequently, there is a need to understand each of the many variables that comprise the BFR exercise stimulus, such as those relating to the cuff application (e.g., pressure, restriction duration etc.) and the exercise prescribed (intensity/loading, volume etc). A review of the literature (Chapter Two) identified that one particular variable receiving little attention is whether blood flow must be restricted during the inter-set recovery periods (continuous BFR), or if this is only necessary during the working periods (intermittent BFR). A greater understanding of such variables will permit the configuration of optimal and evidence-based BFR protocols, meaning that this mode of exercise may be tailored to suit the goals and health considerations of a wide spectrum of individuals.As such, this thesis encompasses a series of independent, yet related studies that examine a wide range of acute and chronic responses to different low-load BFR resistance exercise protocols, with the ultimate aim of contributing towards the current evidence-based and best-practice guidelines for BFR exercise. Study One compared chronic training adaptations between continuous and intermittent BFR protocols. The lack of differences between protocols in several morphological and functional outcomes suggested that the additional metabolic stress imposed on skeletal muscle with a continuous BFR protocol may not be necessary. However, because the study involved untrained participants, the findings may not translate to well-trained and athletic cohorts.Study Two compared differences between continuous and intermittent BFR for a wide range of acute outcomes, including neuromuscular, cardiovascular, metabolic and biochemical responses. The inclusion of trained participants, and a traditional high-load resistance exercise group permitted more ecologically valid comparisons between BFR protocols and the current gold-standard method. It was observed that the maintenance of the BFR stimulus during rest periods significantly altered the intramuscular oxygen kinetics, and as a consequence, was capable of inducing the same magnitude of peripheral fatigue in skeletal muscle as much higher-load exercise. In addition, these differences occurred at no additional cardiovascular demand, thus indicating that this exercise modality is safe compared with traditional approaches. Together, these data suggested that a continuous BFR application may be favourable for presenting a maximal adaptive stimulus for trained individuals. However, an intermittent BFR application induces less fatigue, and may be preferable if subsequent performance takes precedence over adaptation.The final study of this thesis, Study Three, included a comprehensive comparison of low-load resistance exercise continuous BFR, and traditional high-load resistance exercise. Novel aspects of this study involved the use of trained participants, the high-quality assessments of muscle morphology (magnetic resonance imaging and immunohistochemical analysis) and the inclusion of both acute and chronic components. The key findings were that, in trained individuals, the exclusive use of low-loads with BFR did not negatively influence muscular strength or power and produced similar skeletal muscle hypertrophy to traditional high-load training. Acutely, these adaptations were achieved by activation of key intracellular signalling pathways, including the mammalian target of rapamycin (mTOR) and extracellular signal-regulated kinases (ERKs), which were similar to high-load resistance exercise. Together, these findings validate low-load continuous BFR as an effective surrogate to high-load resistance training in trained individuals.This thesis examined the acute and chronic responses to different resistance protocols differing in loading conditions and BFR protocols. The studies presented herein address a prominent gap in the literature on BFR methodology, educating scientists and practitioners on the effects of maintaining vs releasing the BFR stimulus during inter-set rest periods. Moreover, the validation of low-load BFR exercise in trained individuals for stimulating acute anabolic processes within muscle that translate to favourable chronic adaptations has important implications for both clinical and athletic cohorts.

The use of exosomes in clinical settings is progressively becoming a reality, as clinical trials testing exosomes for diagnostic and therapeutic applications are generating remarkable interest from the scientific community and investors. Exosomes are small extracellular vesicles secreted by all cell types playing intercellular communication roles in health and disease by transferring cellular cargoes such as functional proteins, metabolites and nucleic acids to recipient cells. An in-depth understanding of exosome biology is therefore essential to ensure clinical development of exosome based investigational therapeutic products. Here we summarise the most up-to-date knowkedge about the complex biological journey of exosomes from biogenesis and secretion, transport and uptake to their intracellular signalling. We delineate the major pathways and molecular players that influence each step of exosome physiology, highlighting the routes of interest, which will be of benefit to exosome manipulation and engineering. We highlight the main controversies in the field of exosome research: their adequate definition, characterisation and biogenesis at plasma membrane. We also delineate the most common identified pitfalls affecting exosome research and development. Unravelling exosome physiology is key to their ultimate progression towards clinical applications. Video Abstract Supplementary Information The online version contains supplementary material available at 10.1186/s12964-021-00730-1.

With an increasing number of countries and states adopting legislation permitting the use of cannabis for medical purposes, there is a growing interest among health and research professionals into the system through which cannabinoids principally act, the endocannabinoid system (ECS). Much of the seminal research into the ECS dates back only 30 years and, although there has been tremendous development within the field during this time, many questions remain. More recently, investigations have emerged examining the contributions of the ECS to normative development and the effect of altering this system during important critical periods. One such period is adolescence, a unique period during which brain and behaviours are maturing and reorganizing in preparation for adulthood, including shifts in endocannabinoid biology. The purpose of this review is to discuss findings to date regarding the maturation of the ECS during adolescence and the consequences of manipulations of the ECS during this period to normative neurodevelopmental processes, as well as highlight sex differences in ECS function, important technical considerations, and future directions. Because most of what we know is derived from preclinical studies on rodents, we provide relevant background of this model and some commentary on the translational relevance of the research in this area.

Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is involved in lipid-mediated intracellular signalling and the regulation of events such as cytoskeletal reorganization and membrane trafficking. An inducible PI(4,5)P2 depletion system is now available for examining its function in plant cells.

Monoclonal antibodies (mAbs) are a swiftly growing class of targeted therapeutics for malignancies. After their first advent, the antibody (Ab) engineering trail has shown an evolutionary trajectory - from the rodent-derived Abs to the chimeric, humanised and fully human Abs with higher efficacy and lower/no immunotoxicity. Despite possessing great clinical potentials, several reports have highlighted that monospecific mAbs, even with high-affinity, often fail to induce sufficient immunologic responses. The full activation of the immune system demands cooperative interactions of immunotherapies with target antigen (Ag) towards functional avidity. Although the monospecific mAbs show affinity to a target Ag, they often fail to render sufficient avidity necessary for the activation of intracellular signalling mechanisms and the provocation of the immune system. Thus, various Ab/non-Ab scaffolds with much greater therapeutic impacts have been engineered based on the adjustment of their affinity and avidity balance. Novel multivalent Ab scaffolds (e.g. MDX-447, MT110, CD20Bi, TF2 and FBTA05) and mimetic Abs (e.g. adnectin, DARPins and ecallantide) offer improved pharmacokinetic and pharmacodynamic properties. Here, we discuss the avidity and multivalency and provide comprehensive insights into advanced Ab scaffolds used for immunotargeting and therapy of cancer.

Hypoxia has a profound effect on fish behaviour, cardiovascular physiology, most ionic and substrate membrane transport systems, intracellular signalling systems, and gene expression in fish. Extant adult fishes show an extremely wide diversity of form and function, and in many instances this diversity is supported by similarly large variations in cardiovascular anatomy and physiology. Zebrafish offer a number of advantages that allow imaging of the cardiovascular system during development. Temperature has a significant effect on not only the physiology, but also the morphological development of the cardiovascular system during development in zebrafish. Hypoxia has a profound affect on the onset of cardiovascular regulation in that it can accelerate the onset of adrenergic and vasoconstrictive responses relative to the developmental program. Larval fish undergo complex and dynamic patterns of change during their development, and the growth and elaboration of the cardiovascular system is crucial to larval ontogeny.

The sinoatrial node is perhaps one of the most important tissues in the entire body: it is the natural pacemaker of the heart, making it responsible for initiating each-and-every normal heartbeat. As such, its activity is heavily controlled, allowing heart rate to rapidly adapt to changes in physiological demand. Control of sinoatrial node activity, however, is complex, occurring through the autonomic nervous system and various circulating and locally released factors. In this review we discuss the coupled-clock pacemaker system and how its manipulation by neurohumoral signaling alters heart rate, considering the multitude of canonical and non-canonical agents that are known to modulate sinoatrial node activity. For each, we discuss the principal receptors involved and known intracellular signaling and protein targets, highlighting gaps in our knowledge and understanding from experimental models and human studies that represent areas for future research.

Fatty acids play a very important role in our body. Both essential and nonessential fatty acids are necessary for our body. They help in regulation of membrane structure and function, transcription factor activity, regulation of intracellular signalling pathways, and gene expression. Traditional oilseed crops in India include safflower, sunflower, mustard, soybean, etc. Linseed and garden cress seeds are also being popularized due to its healthy effect on the heart and body. The fatty acids are categorized based on bond present in them, viz., saturated (SFA), monounsaturated (MUFA), and polyunsaturated fatty acids (PUFA). As per WHO (Interim summary of conclusions and dietary recommendations on total fat and fatty acids. The joint FAO/WHO expert consultation on fats and fatty acids in human nutrition. World Health Organization, Geneva, Switzerland, 2008) specifications, the ratio between sat-, mono-, and polyunsaturated fatty acids should be 1:1.5:1 which is considered ideal for healthy heart. This exact ratio is not present in any oil. Moreover, the oils are lacking in one or more essential fatty acids. Therefore, the present article shows the brief description and comparison of fatty acid composition of seven different oilseeds, viz., mustard, garden cress, sunflower, safflower, groundnut, linseed, and sesame seeds, to find out among the best oil to be consumed.

Toll-Like Receptors (TLRs) are membrane-bound proteins that recognize invading organisms with Pathogen-Associated Molecular Patterns (PAMPs) and Damage-Related Molecular Patterns (DAMPs)...

Motivation Due to the complexity and heterogeneity of multicellular biological systems, mathematical models that take into account cell signalling, cell population behaviour and the extracellular environment are particularly helpful. We present PhysiBoSS, an open source software which combines intracellular signalling using Boolean modelling (MaBoSS) and multicellular behaviour using agent-based modelling (PhysiCell). Results PhysiBoSS provides a flexible and computationally efficient framework to explore the effect of environmental and genetic alterations of individual cells at the population level, bridging the critical gap from single-cell genotype to single-cell phenotype and emergent multicellular behaviour. PhysiBoSS thus becomes very useful when studying heterogeneous population response to treatment, mutation effects, different modes of invasion or isomorphic morphogenesis events. To concretely illustrate a potential use of PhysiBoSS, we studied heterogeneous cell fate decisions in response to TNF treatment. We explored the effect of different treatments and the behaviour of several resistant mutants. We highlighted the importance of spatial information on the population dynamics by considering the effect of competition for resources like oxygen. Availability and implementation PhysiBoSS is freely available on GitHub (https://github.com/sysbio-curie/PhysiBoSS), with a Docker image (https://hub.docker.com/r/gletort/physiboss/). It is distributed as open source under the BSD 3-clause license. Supplementary information Supplementary data are available at Bioinformatics online.

Motivation Intracellular signalling is realized by complex signalling networks, which are almost impossible to understand without network models, especially if feedbacks are involved. Modular Response Analysis (MRA) is a convenient modelling method to study signalling networks in various contexts. Results We developed the software package STASNet (STeady-STate Analysis of Signalling Networks) that provides an augmented and extended version of MRA suited to model signalling networks from incomplete perturbation schemes and multi-perturbation data. Using data from the Dialogue on Reverse Engineering Assessment and Methods challenge, we show that predictions from STASNet models are among the top-performing methods. We applied the method to study the effect of SHP2, a protein that has been implicated in resistance to targeted therapy in colon cancer, using a novel dataset from the colon cancer cell line Widr and a SHP2-depleted derivative. We find that SHP2 is required for mitogen-activated protein kinase signalling, whereas AKT signalling only partially depends on SHP2. Availability and implementation An R-package is available at https://github.com/molsysbio/STASNet. Supplementary information Supplementary data are available at Bioinformatics online.

The receptor for advanced glycation end products (RAGEs) was first illustrated in the year 1992. RAGE is a single-transmembrane and multi-ligand component of the immunoglobulin protein super family. The engagement of RAGE turns out to an establishment of numerous intracellular signalling mechanisms resulting in the progression and perpetuation of many types of cancer including, the pancreatic cancer. The present review primarily focuses on the multi-ligand activation of RAGEs leading to the downstream signalling cascade activation. The kick start of the RAGEs activation leads to the several anomalies and includes multiple types of cancers. The RAGE expression correlates well with the survival of pancreatic cancer cells leading to the myeloid response. RAGEs assist in the tumourogenesis which enhance and thrive to its fullest in the stressed tumour microenvironment. An improved perceptive of its involvement in pancreatic cancer may offer novel targets for tumour supervision and risk measurement.

Rejeneratif tedaviler progenitor/kok hucreler, doku iskeleleri ve buyume faktorleri olmak uzere uc bilesene dayalidir. Buyume faktorleri sinyal molekulleri olarak davranan polipeptidlerdir ve farklilasma, cogalma ve migrasyonu iceren hucresel olaylari module ederler. Hucresel aktivitelerin duzenlenmesi sirasinda, buyume faktorleri kendilerine ozgu hucre yuzey reseptorlerine baglanirlar ve hucre ici sinyal yolaklari ve transkripsiyon faktorlerini kullanirlar. Transforme edici buyume faktoru, vaskuler endotelyal buyume faktoru, platelet kaynakli buyume faktoru, fibroblast buyume faktoru ve insulin benzeri buyume faktoru pulpa-dentin kompleksinin tamir ve rejenerasyonunda etkilerinin gosterildigi rejeneratif endodontik calismalara konu olan buyume faktorleridir. Bu derlemenin amaci, hucre ici fonksiyonlarda buyume faktorlerinin onemini tartismaktir. Anahtar Kelimeler: Buyume faktorleri, rejeneratif endodonti, pulpa-dentin kompleksi GROWTH FACTORS IN REGENERATIVE ENDODONTICS ABSTRACT Regenerative therapies are based on three components which are progenitor/stem cells, scaffolds and growth factors. Growth factors are polypeptides that act as signal molecules and modulate celullar actions including differentiation, proliferation and migration. During the regulation of cellular activities, they bind specific cell-surface receptors and use intracellular signalling pathways and transcription factors. Transforming Growth Factor, Vascular Endothelial Growth Factor, Platelet Derived Growth Factor, Fibroblast Growth Factor and Insulin-like Growth Factor are growth factors mostly subjected to regenerative endodontic studies in which their effects are displayed on pulp-dentine complex repair and regeneration. The aim of this review is to discuss the importance of growth factors in intracellular functions. Keywords: Growth factors, regenerative endodontics, pulp-dentine complex

Our gut forms an important organ and its formation, functioning and homeostasis are maintained by several factors including cell signalling pathways and commensal microflora. These factors affect pathological, physiological and immunological parameters to maintain gut health and prevent its inflammation. Among these, different intracellular signalling pathways play an important role in regulating gut homeostasis. These pathways are in turn regulated by various microRNAs that play a key role in maintaining the balance between tolerance and inflammation. This review highlights the importance of various cell signalling pathways in modulating gut homeostasis and the role specific miRNAs play in their regulation.

Over the past decades, a number of prolactin receptor (PRLR) antagonists have been developed, which can be divided into two categories, PRLR analogue and anti-PRLR antibody. However, until now, there have been no commercially available PRLR antagonists. Here, we described a new approach for the preparation of PRLR antagonist, namely internal image anti-idiotypic antibody strategy. The hybridoma technique was used to generate anti-idiotypic antibodies to PRL. Competitive ELISA, competitive receptor-binding analysis and immunofluorescence assay (IFA) were then used to screen and characterize anti-idiotypic antibodies to PRL. One internal image anti-idiotypic antibody, termed MG7, was obtained. A series of experiments demonstrated that MG7 behaved as a typical internal image anti-idiotypic antibody (Ab2β). MG7 exhibited effective antagonistic activity, which not only inhibited PRL binding to PRLR in a dose-dependent manner but also inhibited PRLR-mediated intracellular signalling. Furthermore, MG7 also blocked Nb2 cell proliferation induced by PRL. The current study suggests that MG7 has the potential application in the PRL/PRLR-related studies in future. In addition, this work also suggests that the internal image anti-idiotypic antibody may represent a novel strategy for the development of PRLR antagonist.

Pain sensitivity is strongly modulated by intracellular signalling. The analysis of pain signalling is hampered by small numbers of neurons, high cellular heterogeneity, and near absence of transfectability. HCS microscopy allowed us to proof that nociceptive neurons express specific signalling components, to analyze signalling activity of endogenous signalling components with single cell resolution, and thereby to suggest blockers of Nav1.7 as novel therapeutic approach to boost the efficacy of opioids.

The formation and release of neutrophil extracellular traps (NETs) discovered in 2004 by Volker Brinkmann and Arturo Zychlinsky cast a new light on the role of neutrophils in the non-specific immune response of the body. This discovery has resulted in the rapid development of neutrophil studies in different bacterial and autoimmune diseases as well as neoplasms. Research is also being performed on the role of different signalling pathways engaged in the induction of NETosis, a unique form of a programmed cell death leading to the creation of NETs. The literature provides information on the structure and composition of neutrophil extracellular traps. This review presents the latest data on NET formation and the role of their key components, as well as describing the intracellular signalling pathways leading to the generation of NETs that have been discovered.

Objectives Impaired angiogenesis in sciatic nerve is a major complication of diabetic neuropathy. Protein kinase B (AKT) and extracellular signal regulated kinase (ERK) signaling pathways play critical roles during capillary-like network formation in the angiogenesis process. Methods Twenty-four adult male Wistar rats (weight: 250–300 g) were used in the research. The rats were randomly divided into four groups (n = 6): (1) diabetic (Dia), (2) diabetic + castration (Dia-Cas), (3) diabetic + exercise (Dia-E), and (4) diabetic + castration + exercise (Dia-Cas-E). Type 1 diabetes (T1D) was induced with streptozotocin (50 mg/kg). After 6 weeks, sciatic nerve was separated and used for histological evaluation and determination of phosphorylated AKT (P-AKT) and phosphorylated ERK (P-ERK) levels by ELISA method. Results Glucose levels decreased in the Dia-E group compared to the Dia-Cas group (p p p Conclusions Voluntary exercise in diabetic rats increases angiogenesis in the sciatic nerve. The possible mechanism is the increase of P-ERK 1/2 but not P-AKT levels in the sciatic nerve of T1D rats.

In recent years, fluorescent dyes have been frequently used for monitoring mitochondrial membrane potential to evaluate mitochondrial viability and function. However, the reproducibility of the results across laboratories strongly depends upon following well validated and reliable protocols along with the appropriate controls. Herein, we provide a practical user guide for monitoring mitochondrial membrane potential in whole cells using a fluorescent cationic probe. The data analysis of mitochondrial membrane potential we provide is one associated with the impact of xenobiotics such as tobacco smoking on blood-brain barrier endothelial cells including both mouse primary (mBMEC) and a mouse-based endothelial cell line (bEnd3) in a side by side comparison.

In 2013, we discovered that the entitled "calcium paradox" phenomenon, which means a paradoxical sympathetic hyperactivity produced by l-type Ca(2+) channel blockers (CCBs), used in antihypertensive therapy, is due to interaction between the intracellular signalling pathways mediated by Ca(2+) and cAMP (Ca(2+)/cAMP interaction). In 2015, we proposed that the pharmacological manipulation of this interaction could be a new therapeutic strategy for increasing neurotransmission in psychiatric disorders, and producing neuroprotection in the neurodegenerative diseases. Besides the paradoxical sympathetic hyperactivity produced by CCBs, several clinical studies have been demonstrating pleiotropic effects of CCBs, including neuroprotective effects. CCBs genuinely exhibit cognitive-enhancing abilities and reduce the risk of dementia, including Alzheimer's, Parkinson´s disease and others. The molecular mechanisms involved in these pleiotropic effects remain under debate. Our recent discovery that the "calcium paradox" phenomenon is due to Ca(2+)/cAMP interaction may provide new insights for the pharmacological treatment of neurological and psychiatric disorders, including enhancement of current therapies mainly by reducing adverse effects, and improving effectiveness of modern medicines. Whether Ca(2+)/cAMP interaction is involved in CCBs pleiotropic effects also deserves special attention. Then, the pharmacological manipulation of the Ca(2+)/cAMP interaction could be a more efficient therapeutic strategy for increasing neurotransmission in psychiatric disorders, and producing neuroprotection in the neurodegenerative diseases. Thus, in this review we summarize the current knowledge of this field, making new directions and future perspectives.

For over a decade, pepducins have been utilized to develop unique pharmacological profiles that have been particularly challenging for traditional drug discovery methods. It is becoming increasingly clear that these cell-penetrating lipopeptides can access receptor conformations that are currently not accessible through orthosteric targeting. This review addresses the emerging concepts in the development of pepducins including the elicitation of biased signalling, pepducin polypharmacology and recent insight into their mechanism of action.

Although advances in biological medicine have seen significant progress in the treatment of autoimmune and inflammatory disease, many patients do not experience a satisfactory response. Hence, there are two challenges facing the medical research community. The first is to continue development in the field of existing biological therapies, such as monoclonal antibodies. The second is to open new frontiers of research and explore treatment alternatives for non-responders to other therapies. Attention has increasingly turned to the therapeutic potential of small molecule weight kinase inhibitors (SMKIs), currently used extensively in oncology and haematology. Initial research into the therapeutic value of SMKIs for autoimmune and inflammatory diseases has been encouraging. SMKIs are taken orally, which reduces cost for the health provider, and could increase compliance for the patient. This is why research is now focusing increasingly on SMKIs as a new generation line of treatment in these diseases. Tofacitinib, an inhibitor of Janus-kinase, is currently the only drug approved for the treatment of rheumatoid arthritis by FDA. However, much more needs to be done to understand the intracellular signalling pathways and how these might affect disease progression before solid conclusions can be drawn.

Recent studies have shown that silver is no longer restricted to just being found in soil and natural waters. The metal and its various forms are used in medicine as nanoparticles for various applications. After assimilating and reviewing a considerable volume of literature, it can be safely stated that silver displays different extents of bioaccumulation depending on the ecosystem involved. Aquatic ecosystems were revealed to be less prone to the toxic effects of the metal due to the buffering capacity of water and its dissolved contents. Evidence suggests that even though bioaccumulation is possible in terrestrial animals, biomagnification is not. Humans are more prone to bioaccumulation due to the use of silver nanoparticles in medicine. The toxic effects of silver are attributed to the formation of free silver (I) ions (Ag + ). Biochemical effects are brought about by interactions with proteins, deoxyribonucleic acid (DNA) and intracellular signalling pathways. Physiological effects are manifestations of biochemical effects in different regions of the body. Literature has shown that by means of physiological and biochemical effects, silver bioaccumulation gives rise to both benign and harmful disease, namely argyria, cardiovascular disease and respiratory disease.

Background: Medicinal plant and plant products have shown tremendous potentials and are used beneficially in the treatment of inflammation and in the management of diseases with significant inflammatory components. Many medicinal plants employed as anti-inflammatory and anti-phlogistic remedies lack the gastro-erosive side effects of non-steroidal anti-inflammatory drugs (NSAID) or the plethora of unwanted side effects associated with steroidal anti-inflammatory drugs. In order to harness and optimise the applications of these herbs in inflammatory diseases, there is a need to understand how these herbs produce their anti-inflammatory actions. Materials and Methods: This paper is a review of some anti-inflammatory herbs and their molecular mechanisms of action. A literature search and analysis of published manuscript was employed to x-ray research findings that show how medicinal plants produce anti-inflammatory activities. Results: Many studies have shown that anti-inflammatory activities of herbal extracts and herb-derived compounds are mainly due to their inhibition of arachidonic acid (AA) metabolism, cyclo-oxygenase (COX), lipo-oxygenase (LOX), pro-inflammatory cytokines, inducible nitric oxide, and transcription activation factor (NF-κB). Some anti-inflammatory medicinal herbs are reported to stabilize lysosomal membrane and some cause the uncoupling of oxidative phosphorylation of intracellular signalling molecules. Many have also been shown to possess strong oxygen radical scavenging activities. Conclusion: Most of the mechanisms by which anti-inflammatory medicinal plants act are related and many herbal products have been shown to act through a combination of these molecular pathways.

Currently, Systemic Lupus Erythematosus (SLE) therapies range from antimalarials to glucocorticoids, in addition to immunosupressive agents or biologics such as rituximab or belimumab, when needed. Several unmet needs remain in the treatment SLE and more targeted drugs with improved safety profiles are expected. Based on recent advances in the understanding of the complex pathogenesis of SLE, several targeted treatments are currently assessed in clinical trials. In this study, we performed a systematic review of all targeted therapies under clinical development in SLE in 17 online registries of clinical trials. The search yielded a total of 1140 trials, from which we identified 74 targeted therapies for SLE. Those treatments target inflammatory cytokines, chemokines, or their receptors (n = 17), B cells or plasma cells (n = 17), intracellular signalling pathways (n = 10), T/B cells costimulation molecules (n = 8), interferons (n = 7), plasmacytoid dendritic cells (pDC) (n = 3), as well as various other targets (n = 12). Not all these candidate drugs will reach phase III, but the broad spectrum of drugs being investigated may satisfy the urgent need for improved lupus medications. The identification of biomarkers that would allow adequate prediction of response-to-therapy remains high, but when solved will allow a more rationale selection of the optimal pharmacological agent at the patient level.

Because of their antimicrobial properties, the use of silver nanoparticles (AgNPs) is increasing fast in industry, food, and medicine. In the food industry, nanoparticles are used in packaging to enable better conservation products such as sensors to track their lifetime, and as food additives, such as anti-caking agents and clarifying agents for fruit juices. Nanoemulsions, used to encapsulate, protect and deliver additives are also actively developed. Nanomaterials in foods will be ingested and passed through the digestive tract. Those incorporated in food packaging may also be released unintentionally into food, ending up in the gastrointestinal tract. It is therefore important to make a risk assessment of nanomaterials to the consumer. Thus, exposure to AgNPs is increasing in quantity and it is imperative to know their adverse effects in man. However, controversies still remain with respect to their toxic effects and their mechanisms. Understanding the toxic effects and the interactions of AgNPs with biological systems is necessary to handle these nanoparticles and their use. They usually generate reactive oxygen species resulting in increased pro-inflammatory reactions and oxidative stress via intracellular signalling pathways. Here, we mainly focus on the routes of exposure of AgNPs, toxic effects and the mechanisms underlying the induced toxicity.

Ageing is a cellular process with many facets, some of which are currently undergoing a paradigm change. It is the case of “mitochondrial theory of ageing”, which, interestingly, has been found lately to cross paths with another ageing dysfunctional process – intracellular signalling – in an unexpected point (or place) – caveolae. The latter represent membrane microdomains altered in senescent cells, scaffolded by proteins modified (posttranslational or as expression) with ageing. An important determinant of these alterations is oxidative stress, through increased production of reactive oxygen species that originate at mitochondrial site. Spanning from physical contact points, to shared structural proteins and similar function domains, caveolae and mitochondria might have more in common than originally thought. By reviewing recent data on oxidative stress impact on caveolae and caveolins, as well as possible interactions between caveolae and mitochondria, we propose a hypothesis for senescence-related involvement of caveolins.