The primary objective of population-based newborn screening is the early identification of asymptomatic infants with a range of severe diseases, for which effective treatment is available and where early diagnosis and intervention prevent serious sequelae. Primary immunodeficiency diseases (PID) are a heterogeneous group of inborn errors of immunity. Severe combined immunodeficiency (SCID) is one form of PID which is uniformly fatal without early, definitive therapy, and outcomes are significantly improved if infants are diagnosed and treated within the first few months of life. Screening for SCID using T cell receptor excision circle (TREC) analysis has been introduced in many countries worldwide. The utility of additional screening with kappa recombining excision circles (KREC) has also been described, enabling identification of infants with severe forms of PID manifested by T and B cell lymphopenia. Here, we review the early origins of newborn screening and the evolution of screening methodologies. We discuss current strategies employed in newborn screening programs for PID, including TREC and TREC/KREC-based screening, and consider the potential future role of protein-based assays, targeted sequencing, and next generation sequencing (NGS) technologies, including whole genome sequencing (WGS). Population-based newborn screening enables the early identification of asymptomatic infants with a range of severe diseases, for which effective treatment is available and where early diagnosis and intervention prevent serious sequelae. Primary immunodeficiency diseases (PID) are a heterogeneous group of inborn errors of immunity, the majority of which present in infancy and result in significant morbidity and mortality. Until recently, it was not possible to identify infants with PID prior to the onset of clinical symptoms, at which point they typically have complications of severe and protracted infection. Advances in technology have enabled identification of infants with severe forms of PID manifested by T and/or B cell lymphopenia. Here, we review the early origins of newborn screening and the evolution of screening methodologies. We review current strategies employed in newborn screening programs for PID, including T cell receptor excision circle (TREC) and kappa recombining excision circles (KREC)-based screening approaches, and discuss the potential future role of protein-based assays, targeted sequencing, and next generation sequencing (NGS) technologies.Phenylketonuria (PKU), also known as Følling’s disease, was first identified in 1934 by the Norwegian biochemist and physician Ivar Asbjörn Følling (1888–1973), who evaluated two siblings aged 4 and 7 years with developmental delay and an unusual urinary odor . FøllingBacillus subtilis by phenylalanine and associated compounds .T cell receptor excision circles (TREC) are small, circular pieces of episomal DNA which are formed during T cell receptor (TCR) rearrangement in naïve T cells and are thus surrogate markers for recent thymic emigrants. TREC were first visualized by electron microscopy as circular, extra-chromosomal DNA in mouse thymocytes in 1982  and wereAlthough identification of infants with SCID was the intended aim of TREC-based newborn screening programs, it became evident that in addition to this disorder, the assay would also identify infants with T cell lymphopenia due to other primary and secondary causes Table . For exaIn addition to TREC analysis, other SCID screening approaches have been proposed. In Italy, newborns are screened for SCID with the TREC assay, along with tandem mass spectrometry to identify ADA deficiency, which also enables identification of infants with delayed-onset ADA deficiency –33. A siPrimary immunodeficiency diseases were previously thought to be rare entities, and the incidence of SCID was unknown. Prospective screening programs have since enabled the true incidence of SCID to be determined. Furthermore, screening has facilitated the identification and treatment of infants who would have otherwise died from complications of undiagnosed SCID. Based on studies in the USA where over 3 million infants were screened, the incidence of SCID was found to be much higher than expected at 1/58000 and the incidence of clinically relevant T cell lymphopenia was 1/7300 . The annBTK gene) and autosomal recessive XLA-like disorders. Patients have absent B cells, extremely low or undetectable immunoglobulin levels, and an increased susceptibility to severe infection with bacteria and other pathogens [IGK locus results in a Vk-Jk coding joint, followed by rearrangement of the intron RSS and Kde elements with deletion of the Ck exon and enhancers. The coding joint remains present in the genome, whereas the KREC with the corresponding signal joint is excluded as a stable, circularized DNA fragment [Mutations in key genes which are essential for B cell ontogeny give rise to congenital B cell deficiency disorders including X-linked agammaglobulinemia (XLA) , Taiwan, Israel, Qatar, and several Canadian regions, and are due to commence in New Zealand in the near future . ScreeniComplement proteins are an important component of the innate immune system, and deficiencies in complement give rise to a variable clinical phenotype, including autoimmune disease, renal disease, and a susceptibility to specific bacterial infections which frequently result in life-threatening infection . DisordeUNC13D may be identified [UNC13D inversion mutations, and it has been demonstrated that a reduction in the wild type gene copy numbers is an effective way to screen for affected individuals [Targeted DNA sequencing has previously been employed as a screening strategy for selected diseases, such as glutaric acidemia type I and cystic fibrosis. This approach has also been described as a potential method by which infants with familial hemophagocytic lymphohistiocytosis (FHLH) due to mutations in Primary immunodeficiency diseases are a heterogeneous group of disorders, which differ in terms of clinical phenotype, laboratory findings, and underlying molecular abnormalities. There are currently over 300 different genetic mutations associated with PID , and thiThe National Institutes of Health is currently evaluating the role of up-front NGS in newborn screening as part of the NSIGHT  project . Whole-eCFTR mutations [Recently, Pavey et al. used WGS to screen 1349 newborn and parent trios for variants in 329 known PID-associated genes . Applyinutations . Bodian utations .Applying a WGS-based approach to newborn screening represents a change in paradigm, and there are many factors which must be taken into consideration before adopting this approach in population-based screening programs , 58. FirManagement of PID is variable and is dependent on the disease, clinical phenotype, and molecular defect. In the case of SCID, HSCT or gene therapy is currently available curative therapies. Advances in CRISPR-Cas based genome editing have led to interest in potential therapeutic applications of this modality. Scott and Zhang recently highlighted that prior to institution of CRISPR-based therapy, patients should be screened using WGS to ensure safety and reduction of off-target effects . The earFGFR3 mutations which give rise to achondroplasia can also be identified. Targeted or whole genome sequencing of the fetus is also possible, along with sequencing of the fetal transcriptome and methylome, enabling identification of a wide range of disorders [Newborn screening enables identification of infants with key disorders requiring early intervention soon after birth. Prenatal screening for disorders such as Trisomy 21 and neural tube defects is well established in obstetric practice, along with prenatal diagnostics in the case of a high risk of a specific genetic disorder based upon positive family history or other factors. However, this has typically required invasive procedures which carry associated risks, such as amniocentesis and chorionic villus sampling. Technological advances have resulted in the ability to detect fetal DNA in maternal plasma, and Lo et al. demonstrated that massively parallel sequencing (MPS) enables detection of fetal aneuploidies such as trisomies 21, 13, and 18 . In addiisorders . PrenataNewborn screening practices worldwide have evolved significantly since the discovery of PKU in 1934, with rapid development of new and improved assays to enable identification of a wider range of conditions in infants and facilitating early diagnosis and treatment, and improving clinical outcomes. Primary immunodeficiency diseases are a heterogeneous group of disorders, and no single assay at the present time will identify all forms of PID, necessitating a challenge of current newborn screening paradigms. In the genomic era, it is likely that this will involve up-front next generation sequencing, including whole exome sequencing and ultimately, whole genome sequencing. This approach must be evaluated in large, prospective trials prior to adopting this strategy in population-based newborn screening programs.


Nanobiotechnology has the potential to revolutionize diverse sectors including medicine, agriculture, food, textile and pharmaceuticals. Disease diagnostics, therapeutics and crop protection strategies are fast emerging using nanomaterials preferably nanobiomaterials. It has potential for development of novel nanobiomolecules which offer several advantages over conventional treatment methods. RNA nanoparticles with many unique features are promising candidates in disease treatment. The miRNAs are involved in many biochemical and developmental pathways and their regulation in plants and animals. These appear to be a powerful tool for controlling various pathological diseases in human, plants and animals, however there are challenges associated with miRNA based nanotechnology. Several advancements made in the field of miRNA therapeutics make it an attractive approach, but a lot more has to be explored in nanotechnology assisted miRNA therapy. The miRNA based technologies can be employed for detection and combating crop diseases as well. Despite these potential advantages, nanobiotechnology applications in the agricultural sector are still in its infancy and have not yet made its mark in comparison with healthcare sector. The review provides a platform to discuss nature, role and use of miRNAs in nanobiotechnology applications. Nanotechnology, an emerging field of research encompassing the creation and usage of materials at nano scale, at the interface of physical and biological sciences using either top-down approaches or bottom-up assembly. In the biological domain a huge number of well-organised structures and nanomachines made up of macromolecules have been designed to carry out various biological functions. Their fascinating make-up have encouraged many biometric designs due to unique characteristics of DNA, RNA and proteins at nano scale, these can be a platform for the development of nano structures and devices. The idea was pioneered 30 years ago by developing nanostructures using DNA, leading to explosion of information in the well-recognised field of DNA nanotechnology . NowadayThe miRNA is endogenous non-coding RNA, which recently has been found to play a key role in regulatory functions responsible for gene expression in all the organisms. Typically, miRNA is 18–23 nucleotides in length and plays an important role in post transcriptional regulation. These are active and dynamic in cytoplasm, in spite of the fact that their synthesis and transcription takes place in the nucleus. The miRNA profile is changed in diseased state making miRNAs a great target for drug therapy through RNA nanotechnology. Restoration of down-regulated miRNA or restraint of overexpressed miRNA to return miRNA to its ordinary state is the premise of miRNA based disease control which can be additionally investigated to control and manage other various cellular functions also. The therapeutic potential of miRNAs was first realized with the discovery that downregulation of miR-15 and miR-16 is associated with development of B cell leukaemia [The biogenesis and maturation of miRNA requires different strides which includes nuclear processing, nuclear export and cytoplasmic processing. RNA polymerase II first transcribes the miRNA genes leading to the formation of long primary transcript having a hairpin structure –6. The pPlant miRNAs (highly variable in length) are generally transcribed by RNA pol II in a way similar to animals –25. HoweMiRNAs are involved in various biological processes such as cell growth and development, biogenesis, signal transduction and apoptosis. MiRNA genes are frequently subjected to genetic alterations as they are located in fragile genomic regions. The epigenetic changes can bring about varied miRNA expression prompting to over expression or down regulation of miRNA leading to various diseases like cardiac, diabetes, neurological and many more. MiR-145 is differentially expressed in various types of cancers like colon, breast, lung and prostate –35.P. trichocarpa when infected with stem canker pathogen [Verticillium dahlia in Eggplant [Exserohilum turcicum in maize. They also found that miR811, miR829, miR845 and miR408 were differentially expressed in infected plants [Papaya meleira virus was increased in response to very low levels of PMeV titre and decreased as the viral titre increases [Ziziphus jujube, 12 miRNAs were upregulated two-fold including miR156a, miR156b, miR156c, miR156d, miR156e, miR156 h, miR159e, miR319a, miR395a, miR395b, zju-miRn23 and zju-miRn24. Ten miRNA were down regulated two-fold including miR159a, miR172, miR2111, miR2950, miR399, miR477, miR858b, zju-miRn2, zju-miRn8 and zju-miRn16 in response to Witches’-Broom phytoplasma infection [F. Oxysporum [A large number of miRNAs have been reported in plants, which are evolutionary conserved and regulate several biological processes. These are also found to be associated with various bacterial, fungal and viral diseases. These miRNAs are differentially expressed in growth and development as well in response to biotic and abiotic stress. The miR156 and miR159 are dominant families which are associated with pathogen response in plants and can be targeted for controlling serval bacterial, fungal and viral diseases in plants. These miRNA families may provide an excellent opportunity for engineering disease resistance for crop protection. For example, differential expression of miRNA has also been observed in diseased plants. Twelve miRNAs  were found upregulated in stem bark of pathogen . Many hiEggplant . Gao et Eggplant . Wu et ancreases . In Zizinfection . A decrexysporum . This diPuccinia [R. solanacearum pathogenicity in Arabidopsis [Interplay of TFs with miRNA is well-known. For example in wheat, interaction between miR164 and a novel transcription factor NAC21/22 is said to be involved with resistance against Puccinia . In PopuPuccinia . WRKY trPuccinia . NF-YA tbidopsis .MiRNAs are key governors of altered gene expression under many pathological conditions in plants, animals and humans, and are characterized by altered miRNA levels. Therefore, precise and sensitive assessment of miRNA can lead to early and correct disease diagnosis and make these small noncoding RNA transcripts a valuable biomarker . Convent2 [Gold nanoparticles promise a viable and sensitive detection technology as their optical properties can easily be tuned and can be surface functionalized using a variety of well characterized chemical moieties that is thiols, disulphides and amines . Gold na2 . Carbona2 . The ide2  and nanoNanopore provides a sore single molecule platform for digging into a large variety of life science troubles –97. NanoDespite the fact that we have just barely started to pick up bits of knowledge into the full scope of biologic elements of miRNA, their inclusion in the onset and movement of diseases has created huge enthusiasm for its use in therapeutics. Mounting proofs recommend that miRNA-based treatments, either by restoring or repressing miRNAs expression and activity hold great promise in therapeutics. The unusual expression of miRNA in various diseases makes it a critically important focus for researcher to create more strategies to develop newer techniques for disease diagnostics and treatment. Biologists have understanding that several different miRNAs are upregulated upon infection with various diseases. Under this situation, antisense inhibitors of the overexpressed mature miRNA sequence can be used to hinder the overexpression as restorative technique. The miRNA expression can be reset to restore the normal cell functioning by providing mature miRNA exogenously. Presently, miRNA activity is regulated by two approaches (i) restoring miRNA function using synthetic double stranded miRNAs, (ii) inhibiting miRNA function using chemical modified anti-miR oligonucleotides .The normal functionality of miRNA in the diseased conditions which caused downregulation of miRNA can be regained by two ways . First bWhen miRNA upregulation leads to disease symptoms, the miRNA inhibition therapy can be employed to regulate the expression. Most methods involve miRISC complex disruption. MiRNA inhibition can be done either by using miRNA sponges or antisense oligonucleotides called anti-miRs. In miRNA sponge, the function of a specific miRNA or miRNA family is blocked by transgenic overexpression of RNA molecule harbouring complementary sites to miRNA of interest . ChemicaWith the advancement of RNA innovation, miRNA have drawn extraordinary consideration and their applications in research and pharmaceuticals are being quickly expanded. In addition to repressing gene expression, miRNA can also activate genes in some cases or even bring pseudo activity by themselves.MiRNA or other RNA-based nanostructures, for instance, anti-miRNA oligonucleotides (AMO) have short half-life and immediately degraded in natural fluids by widespread nucleases. Particularly, nanostructures are more sensitive to natural changes, such as the change from in vitro to in vivo conditions.In spite of the fact that pRNA has been shown to be a powerful nano-vector for delivery of miRNA, regardless it faces many difficulties, for example, industry scale synthesis and purification is not all that simple yet.MiRNA activity depends on imperfect base-matching with target sequences, which may lead to lower specificity. Also, a miRNA may focus on a few obscure qualities in the meantime and one gene can be regulated by a few miRNAs , which iOligonucleotides like siRNA, anti-miRNA and synthetic miRNA mimics can be linked with nanocarriers which improves their resistance to nucleases . The chaIn the recent years, nanotechnology has been through many advancements both in terms of novel material development  and applN-acetyl-d-glucosamine and d-glucosamine. It is generally produced by deacetylation of chitin from crustacean shells. Due to its cationic nature, natural abundance, low toxicity, biodegradability and mucoadhesivity [Chitosan is a natural polysaccharide of β-1-4 linked hesivity , 128 it hesivity –131 and hesivity . Deng ethesivity . Louw ethesivity .Liposome mediated DNA/RNA/miRNA delivery is usually done with the help of cationic lipids (CLs) , 136. ThNucleic acid carriers designed with the help of proteins and peptides are widely explored to deliver nucleic acids and small non-coding RNA molecules because the nucleotides are condensed through electrostatic interactions with the positive charge of amino acids. Furthermore, amino acids add to endosomal escape, targeted delivery and bioreversible polyplex stabilization of system . Oh et aAptamers are 3-D  single chain DNA or RNA molecules that bind to a particular target molecule with high affinity and particularity . RepeateInferable from nano-measurement size to volume proportion and its strength, inorganic  nanoparticles are as a rule broadly utilized as promising quality bearers in different biomedical applications. Inorganic nanomaterials have remarkable electrical and optical properties, biocompatibility and in addition low cytotoxicity .Among the different metals, suitability of gold as nanoparticles is highest due to its inactive nature, ease of synthesis, high functionalization, ability, higher absorption coefficient, simplicity of location and potential ability of targeted capacity. Therefore, it is widely utilized for different applications including medication and gene delivery. Due to its remarkable stability, large surface area, surface modification and high biocompatibility gold nanoparticles can retain the native structure and enzymatic activity of the attached proteins or enzyme in the drug delivery. The instability and poly anionic nature of naked miRNAs impede efficient cellular uptake and reduce half-life. Viral delivery requires cloning miRNA mimics/inhibitors requiring costly modifications and both require toxic lipofection or electroporation. To address these challenges, unmodified miRNAs were delivered using cysteamine-functionalized gold nanoparticles (AuNPs) showing highest payload, lowest toxicity, fastest endosomal escape and increased half-lives . Gold naGraphene is also an attractive nano-materials because of its optical, thermal and electrical properties . The nanQuantum dots (QD) are semiconductor-based, monodisperse, nanometer-sized crystals , which cSilica has also been used for miRNA delivery. Mesoporous silica nanoparticles have large surface area for functionalization and tunable pore size which makes it novel and ideal biomaterial for different therapeutic applications . These hBoth natural and synthetic polymers show variation in their structure, molecular weight together with the chemical properties of the repeat units(s), affect their physico-chemical properties. Due to the large diversity in polymers, polymer based nanomaterials have drawn interest as potential delivery vectors. Polymer mediated delivery provides unique advantage over liposome because of their stability in biological fluids after systemic administration. These are much suitable for industrial scale up as they require less time of preparation. Synthetic polymers include dendrimers, polyethyleneimines and polyphosphoesters.It is most extensively studied polymer for use in gene delivery. PEI is utilized for delivery of nucleic acid to the cells because of its buffering limit . It is aDendrimers are mono disperse, nanosized, highly branched macromolecules which are chemically synthesized in a controlled manner to obtain varied surface functionalities. These are important class of polymers used in delivery of oligonucleotides. They have high drug loading capacity either by encapsulation or conjugation. Delivery of mir-7 to human brain glioma cells (U251) using PAMAM dendrimers had higher transfection efficiency and improved therapeutic effects in vitro and in vivo were achieved . The co-co-glycolic acid) PLGAs are a family of water-insoluble polymers that have been widely used in biomedical applications [co-glycolic acid) are safe, biodegradable, and they have a long history of success in enhancing the delivery of therapeutic agents. Reduction in tumour growth was observed by polymer (PLGA-b-PEG) mediated delivery of anti-miR-10b and anti-miR-21 in mice [Poly technology in 1998 and has created boulevard for the treatment of various deadly diseases like cancer . In RNAi159 and amiR-Hc-Pro159 by Niu et al. [Arabidopsis thaliana miRNA159a, miRNA167b and miRNA171a as backbone. Transgenic tobacco plants showed resistance against PVY and PVX [Nicotiana benthamiana. Triple amiRNA expressing transgenic lines showed complete resistance against WSMoV [Like animals, the RNA nanoparticle based miRNA delivery technique can be utilised in plants to target those biotic and abiotic stresses against which there is no effective strategy for combating them. RNA nanotechnology may revolutionize the crop improvement programmes which will be a step towards food security worldwide. The artificial microRNA (amiRNA) technology can target endogenous miRNA precursors to give rise small RNA guiding gene silencing in plants and animals –194. Oliu et al. . AmiRNA u et al. . AmiRNA  and PVX . In whea and PVX . Arabidost WSMoV . EndogenConstruction of RNA nanoparticle involves coupling of functionalities, module joining, subunit labelling and chemical modification of nucleic acid molecules. Both chemical and enzymatic approaches are utilized in synthesizing RNA building blocks. Although the technology has gained much advancement, still there is a scope for improvement.One of the major challenges in RNA nanotechnology is to know the correct structure of the folded RNA nanoparticle. Because of unusual folding properties, the rules that clarify folding of RNA molecules are yet to be classified. With the use of RNA 2D structure prediction programme, only 70% of structure can be predicted accurately , 213. Th2 on C2′ carbon [RNA in its native form is sensitive to nucleases (RNases). This instability has become a limited factor for its application in nano material construction. With the advancement in technology, several steps have been taken to increase the stability of RNA molecule which includes; chemical modification of bases (5-Br-Ura and 5-I-Ura); alterations in phosphate linking  and changes like 2′-F, 2′-O-Me, 2′-NH′ carbon , 218. RN′ carbon , 220. Am′ carbon .In vivo retention time of the nanoparticle is determined by its size making it a critical factor in RNA nanotechnology. Scientific reports suggest that 10–100 nm range particles are optimal for non-viral vector based delivery. Particles within this range are large enough to be retained in the body and small enough to bind the cellular receptors and can cross cell membrane . Hence oPresently, RNA nanoparticles have limited carrying capacity and is a major problem associated with today’s RNA nanotechnology. Only the terminal end of the nanoparticle can be labelled with the molecule, particularly of smaller size. Whole chain labelling can increase the carrying capacity of the nanoparticle but may lead to misfolding of the nanoparticle due to steric hindrance ultimately leading to release of the target molecule. Computational methods can be employed to locate the site where the target molecule can be introduced without disrupting the nanoparticle. The carrying capacity of RNA nanoparticles can be increased by constructing dendrimers and RNA dendrimers have been developed up to generation 4 . DendrimHigh cost of production and purification of RNA nanoparticles has restricted its use in clinical applications. Typically, RNA nanoparticles are mainly designed within the range of chemical synthesis (maximum of 80 nt). With the advancement in the technology, the cost of oligosynthesis is significantly reduced. Now 10 g of RNA can be synthesized per cycle. However, large scale purification is still a problem. Gel electrophoresis and high-performance liquid chromatography have limited capabilities giving low yield. Recently, ultracentrifugation has been employed for purification of RNA nanoparticles with high yield .Receptor mediated endocytosis is one of the way through which RNA nanoparticles can be delivered in human body. Thus, tracking of intracellular RNA nanoparticle is another major challenge with respect to RNA nanotechnology. Similar to endocytosis pathway, RNA nanoparticles first forms the early endosomal vesicle. After sorting, RNA nanoparticles are shifted to late endosomes and then to lysosomes, where these particles get entrapped without reaching the target. An 8 nt, anti-miR LNA fragment was successful in endosomal escape in RNA nanoparticle mediated cancer therapy , but theWith the natural occurrence of miRNAs and their role in disease detection and prevention, these small RNAs prove to be a brilliant possibility for use as novel therapeutics. With effective delivery of miRNAs to diseased cells, regulation of specific gene expression and modification of cellular function can be accomplished however successful detection, targeting and delivery of the miRNAs are required. Several distinctive methodologies have been utilised for the delivery of miRNA and different detection procedures are employed using different nanoparticles. However, limitations associated with miRNA delivery can be encountered using RNA nanotechnology, which aims at developing RNA nanoparticles containing a targeting molecule and additionally miRNA. The self-modelling quality of RNA can be used as a powerful technique to design and structure RNA nanoparticles through the interchange of biological, chemical, physical and computational sciences. Inspite of difficulties associated with RNA nanotechnology, this emerging field has an extraordinary application in the field of medicine, biotechnology, synthetic biology and nanotechnology, especially in the field of therapeutics where it can be utilized in the treatment of savage diseases like malignancy and viral ailments. Nanotechnology works with smallest possible particles which raises hopes for improving agricultural productivity. It has also been suggested that artificial miRNA poses fewer biosafety or environmental problems when applied to agriculture compared with transgenic breeding. Nanostructured formulation through mechanisms such as targeted delivery or slow controlled release mechanisms and conditional release, could lead to precision release of active ingredients in response to environmental triggers. RNA nanotechnology in plant science holds promise to detect and control various biotic and abiotic stresses through use of miRNA based biomarkers which will revolutionize crop protection. RNA nanotechnology will help to develop new area of research developing biosensors for forecasting and early detection of plant disease.High thermodynamic stability and amazing diversity of RNA and its structure and function holds great promise over DNA Nanotechnology. Since it can be chemically modified, it has been efficiently utilized in human and animal therapeutics, however, plant nanotechnology is still an emerging area of research and its applications in plant disease detection and protection is yet to be seen. Nanotechnology assisted strategies will play key role in integrated pest management supplementing conventional methods in view of environmental concerns. Plant transformation has been used in improving disease resistance in crop plants using overexpression of disease resistance genes, RNAi etc. Synthetic miRNAs can be of particular use and provide excellent choice in these approaches. Delivery of synthetic miRNA in plants is a major concern but recently clay nanoparticles have been used in delivering miRNA for viral protection. The making of synthetic RNA is not cost effective at present; the new developments in this field may lead to novel cost effective strategies. RNA nanoparticles and their use as spray over plants may be advantageous as it is biocompatible, biodegradable and non-toxic making this approach highly flexible and safe.Nanoparticles synthesized from plants extracts can also be utilized for direct delivery of synthetic miRNA as an efficient method in comparison to Agrobacterium and other direct gene transfer methods in plants. Nanotechnology assisted targeting of miRNA can help in mitigating pests and abiotic stresses under climate change scenario. The miRNAs and transcription factors, being the regulatory molecules, promise to target multigenic traits since these regulatory networks overlap.Rapid and early detection of miRNA responsible for disease in plants is still a challenging task. The miRNA can be detected using nanoparticle tags in diseased state using miRNA assay. The association of electrocatalytic nanoparticles tags to hybridized miRNA molecules generates measurable current upon adding substrate to the solution providing better detection limit over optical miRNA assay. There could be new innovative approaches as well for miRNA detection and their use in therapeutics in humans and animals and crop protection as well in near future.


While basic and clinical research over the last several decades has recognized a number of modifiable risk factors associated with cardiometabolic disease progression, additional and alternative biological perspectives may offer novel targets for prevention and treatment of this disease set. There is mounting preclinical and emerging clinical evidence indicating that the mass of metabolically diverse microorganisms which inhabit the human gastrointestinal tract may be implicated in initiation and modulation of cardiovascular and metabolic disease outcomes. The following review will discuss this gut microbiome–host metabolism axis and address newly proposed bile-mediated signaling pathways through which dysregulation of this homeostatic axis may influence host cardiovascular risk. With a central focus on the major nuclear and membrane-bound bile acid receptor ligands, we aim to review the putative impact of microbial bile acid modification on several major phenotypes of metabolic syndrome, from obesity to heart failure. Finally, attempting to synthesize several separate but complementary hypotheses, we will review current directions in preclinical and clinical investigation in this evolving field. Both genomes have co-evolved over millennia with the microbiome expressing a genetic diversity greater than that of the host [13 . Such se13 . Thus, wN-oxide, a metabolite that correlates with increased risk of cardiovascular disease . Desp. DespN-ose  through heart failure with preserved ejection fraction to atherosclerotic disease and complications of stroke, myocardial infarction and peripheral vascular disease. This review pulls from the modest cohort of randomized controlled trials currently available, which is supported by preclinical mechanistic data and clinical observational studies of bariatric surgery patients. We aim to put into context the most current knowledge on how the microbiome, through bile acid manipulation, has the potential to impact on host cardiovascular and metabolic health in a manner which outlines not only pathways, but also the potential environmental aspects that contribute to this tipping-point and potential novel therapies that might derive from it.Bile acids have systemic applications that far surpass their mechanical role as intestinal detergents. The de novo synthesis of bile acids from cholesterol occurs in hepatocytes through two pathways, termed classical and alternative, which are facilitated by cytochrome P enzymes CYP7A1 and CYP27A1, respectively. In a homeostatic state, bile acids are recycled 4–12 times per day, with less than 5% of secreted bile acids being excreted. However, bile modification by members of the gut microbiome can greatly influence the efficacy at which these compounds are reabsorbed and recycled. Naïve conjugated bile salts, secreted into the descending duodenum, undergo several microbial modification reactions, the first of which involves deconjugation to primary bile acids by bile salt hydrolase (BSH) enzymes. This process involves the removal of an amine group, either taurine or glycine, from the conjugated bile salt. Bile salts can be highly cytotoxic to microorganisms, these enzymes therefore likely evolved as a detoxification mechanism and as such, are expressed in abundance by the major microbial phyla found in the human gastrointestinal tract . This reThe gut microbiome potentially represents a “bounty or burden” in terms of its metabolic capacity and how this impacts upon host cardiometabolic health. In line with this, the variety of microbes that humans host appear to have influence over virtually every metabolic process; altering adiposity, liver lipid metabolism and glucose metabolism, as well as heart and vascular function . CurrentObesity represents a central facet of the metabolic syndrome and a preponderance of microbiome–host metabolism interaction studies have focused on it. One of the most important early clinical discoveries was that perturbations in the gut microbiome caused by antibiotic administration during early development can have lasting effects on both the composition and richness of the microbiome , with deSome of the most convincing clinical data to indicate a role of the gut microbiome in obesity has arisen from bariatric surgery studies. The mechanism underlying efficacy of these obesity-curbing surgeries, including vertical sleeve gastrectomy and Roux-en Y gastric bypass, has long puzzled researchers. Observational clinical studies have catalogued the gut microbiome alterations observed in these patients, consistently reporting increased Proteobacteria and decreased Bacteroides levels . In addiIn terms of the weight attenuating effects observed in mice, several groups have now pointed to the potent endogenous FXR antagonist, TβMCA, as a key modulator of host adiposity. The Wallenberg Laboratory initially identified bile acid enrichment in the enterohepatic system of germ-free mice, demonstrating additionally the regulation of bile acid synthesis through the FXR-FGF15 axis . The samIntriguingly, bile acids have also shown the propensity to impact upon mammalian energy expenditure through the thyroid system. Watanabe and colleagues first demonstrated this effect in mice through application of the Takeda-G-protein-receptor 5 (TGR5) agonist, cholic acid (CA) . The effPrevotella copri and Bacteroides vulgatus, has previously been associated with metabolic dysfunction and future insulin resistance [The gut microbiome has been proposed as a highly sensitive and specific tool for the identification of insulin insensitivity –44, althsistance , 48.The potential association of bile acid profile with diabetic state has been acknowledged for decades , yet theInterestingly, TGR5 has been shown to modulate secretion of the incretin GLP-1 , a gut hBacteroidales and Clostridiaceae.There is an established link between the composition and functionality of the gut microbiome and host lipid metabolism , 64. A rLactobacillus reuteri NCIMB 30242, has repeatedly been shown to modestly reduce circulating non-HDL-C levels, as well as other inflammatory markers of atherogenesis [At the center of microbial bile metabolism are the BSH enzymes, which deconjugate bile salts, thereby rendering them exposed to numerous additional modifications. Joyce et al. previously demonstrated the ability of a recombinant BSH-expressing microbe to substantially alter the host circulating bile acid profile and, in turn, markers of host lipid metabolism and circadian rhythm . In suppogenesis , 68. Furogenesis , can benogenesis . This saogenesis .From a clinical standpoint, clean receptor agonists/antagonists would be more attractive than enzyme-secreting microbes, in terms of ease of use, efficacy and safety. In line with this, several bile acid receptor agonists/antagonists have been developed in recent years in an attempt to target this microbial–host crosstalk in certain metabolic pathologies . FXR hasObeticholic acid (OCA) is a highly potent semi-synthetic derivative of the endogenous FXR-agonist chenodeoxycholic acid, which has demonstrated great promise in the treatment of a number of hepatic diseases . The comBeyond lipid metabolism modulation, there is an emerging relationship between gut microbiome status and vascular diseases, such as atherosclerosis, myocardial infarct and stroke , 80. MorBeyond the modification of lipid risk factors, bile signaling has been relatively underexplored as an immunomodulatory pathway directly impacting upon vascular function. Despite this, bile acid receptors have been identified in the vasculature  and sevePreclinical studies with the synthetic TGR5 agonist 6alpha-ethyl-23(S)-methylcholic acid  now also indicate a potential role of TGR5 in the prevention of atherosclerosis. The results demonstrated direct inhibition of NF-κB, which accompanied reduced lipid loading of macrophages and intraplaque inflammation . In termThe risk of CVD events in patients with concomitant hypertension is estimated to be raised by two to threefold ; therefoFrom a wider CVD risk and heart failure-preserved ejection fraction (HF-PEF) perspective, there exists a clear relationship between hypertension, atrial structural change and atrial fibrillation. Given that altered serum bile acid composition is known to induce atrial arrhythmias in patients with atrial fibrillation , it is tWhen considering microbiome effects on bile acid signaling it is noteworthy that physiological differences and inconsistencies exist between animal models and the human system. While the gut microbiome can appear relatively similar between rodents and humans at the level of phylum, further probing to the genus level reveals a juxtaposition in the proportions of several bacterial taxa considered of importance to host health and disease . Diet isA further consideration, which is highly pertinent to this review, is the differences in serum bile profiles between rodents and humans . Most noDespite the outlined successes of several FXR mediated therapeutics, significant disparities in the downstream actions of FXR signaling have been observed . Indeed,Over the past several decades, a multiplicity of clinically relevant functions of the microbiome have become evident including inter alia prevention of enteral colonization by pathogens, education of the host immune system, modulation of host metabolism, bile acid deconjugation and caloric salvage, production of essential short chain fatty acids, vitamin B and K synthesis and even participation in cardiac drug metabolism , 119. AsIntriguingly, microbiome research is now presenting new insights into the multifactorial mechanisms through which cardiometabolic pathologies arise and in doing so presents exciting prospects for development of novel therapeutics and targets within human subjects. These therapies span the spectrum of prebiotic , traditiMoreover, while statin-based therapies represent the gold standard and have proven highly effective in the management of LDL-C and CVD risk, there remain a subset of at-risk hyperlipidemic patients who are partially or entirely intolerant of this drug class. Therefore, there remains scope for in silico mining and development of novel microbial-derived molecular targets , or evenAlready, a number of human studies have indicated promising correlations and direct effects of specific microbial taxa on weight loss, glucose metabolism and correction of other phenotypes of metabolic syndrome , 129–132Although the promise of the personalized medicine predicted upon completion of the Human Genome Project  has not Finally, we must continue to recognize the power of diet and lifestyle adjustments in augmenting a patient’s disease risk status. In line with this, clinical evidence that implicates diet  and exerThe gut microbiome represents the physiological ‘system’ which is perhaps most readily modified by external factors, such as diet , 145 and


POEMS syndrome is an acronym for polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy and skin changes, which is a rare paraneoplastic disease of monoclonal plasma cells. A mandatory criterion to diagnose POEMS syndrome is the presence of a monoclonal plasma cell dyscrasia in which plasma cell leukemia is the most aggressive form. Early identification of the features of the POEMS syndrome is critical for patients to identify an underlying plasma cell dyscrasias and to reduce the morbidity and mortality of the disease by providing early therapy. We present a case of a 64-year-old male who presented with non-specific symptoms and was found to have primary plasma cell leukemia, which was part of his unfortunate POEMS syndrome. Polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome, also known as Crow-Fukase syndrome is a rare paraneoplastic disease of monoclonal plasma cells which was first reported in 1956 . The POEA 64-year-old Caucasian male with a past medical history of hypertension, diabetes mellitus, peripheral neuropathy and recently diagnosed primary hypogonadism presented due to a two-week history of gradual onset of shortness of breath at rest, fatigue and weight loss of 30 lbs in the last one month. He also reported worsening numbness of the lower extremities up to the abdomen despite excellent diabetic control. He denied chest pain, palpitations, cough, orthopnea, paroxysmal nocturnal dyspnea, lightheadedness, fevers, chills or change in stool or urinary symptoms. Social history was significant for only a smoking history of 14 pack years. The physical exam including vital signs was normal, except for splenomegaly and sensory neuropathy of the lower extremities up to the lower abdomen. 9/L) on the differential cell count and thrombocytopenia (platelets of 129 x 109/L). The complete metabolic profile showed an increased corrected calcium, aspartate amino transferase (AST), amino alanine transferase (ALT), alkaline phosphatase and total bilirubin (predominantly direct). Lactate dehydrogenase (LDH) level was normal but β2-macroglobulin and plasma vascular endothelial growth factor (VEGF) was elevated. A chest x-ray showed subacute fractures of multiple ribs , normal white blood cell count but with plasmacytosis  replaced with lenalidomide because of peripheral neuropathy, cisplatin, adriamycin, cyclophosphamide, and etoposide. His course was complicated with liver failure from plasma cell infiltration and subsequent neutropenic fever, septic shock with multi-organ failure. Unfortunately, the patient expired three weeks after diagnosis.9/L. Prognosis is poor due to the high proliferative index, high tumor burden and rapid clinical course of the disease. The estimated median survival is estimated to be only four months [Plasma cell leukemia (PCL) is the most aggressive plasma cell proliferative disorder. Primary PCL occurs de novo in patients with no evidence of previous multiple myeloma, whereas secondary PCL is an evolution of an underlying multiple myeloma. The incidence of PCL is between two percent to four percent of patients with multiple myeloma . It is dr months , therefoPOEMS syndrome is a rare paraneoplastic syndrome due to an underlying plasma cell disorder with an estimated prevalence of 0.3 per 100,000 people . Other nThere are no published randomized trials among patients with POEMS syndrome and therefore treatment is based mainly on case reports and case series. Treatment is directed at the underlying monoclonal plasma cell disorder rather than anti-VEEF antibodies and whether patients have localized monoclonal disease versus those with disseminated bone marrow involvement. Localized plasma cell disease in the form of plasmacytomas without disseminated bone marrow involvement is treated with radiation . With diPOEMS syndrome should be considered when a unique constellation of symptoms is observed, especially when they accompany other nonspecific symptoms such as polyneuropathy, endocrinopathies, and organomegaly. Early identification of the features of the POEMS syndrome is critical for patients so that it can identify an underlying plasma cell dyscrasias and reduce the morbidity and mortality associated with the disease. As seen in our case, plasma cell leukemia which remains the most aggressive plasma cell proliferative disorder was diagnosed with POEMS syndrome being the only identifying feature.


The lateral hypothalamus is comprised of a heterogeneous mix of neurons that serve to integrate and regulate sleep, feeding, stress, energy balance, reward, and motivated behavior. Within these populations, the hypocretin/orexin neurons are among the most well studied. Here, we provide an overview on how these neurons act as a central hub integrating sensory and physiological information to tune arousal and motivated behavior accordingly. We give special attention to their role in sleep-wake states and conditions of hyper-arousal, as is the case with stress-induced anxiety. We further discuss their roles in feeding, drug-seeking, and sexual behavior, which are all dependent on the motivational state of the animal. We further emphasize the application of powerful techniques, such as optogenetics, chemogenetics, and fiber photometry, to delineate the role these neurons play in lateral hypothalamic functions. Another group identified the same peptides via ligand screening of multiple orphan G-protein coupled receptors (GPCRs) using a cell-based reporter system, and, because they promoted feeding, named them “orexins”  has been delineated as a key region for the integration and regulation of sleep, feeding, stress, energy balance, reward, and motivated behavior. Further in-depth analyses of molecular, cellular, and circuit-level functions of the LHA have made progress in gaining a holistic understanding of how the LHA integrates sensory information to influence arousal and behavior. The lateral hypothalamic cell population that has been the most intensely studied thus far is a group of neurons expressing the neurotransmitters hypocretin-1 and -2 . Cells expressing these peptides span the lateral and perifornical/medial hypothalamus . These cells were identified by two groups at essentially the same time. The first approach used directional tag PCR subtraction, a technique adapted to enrich mRNAs that were specifically expressed in the hypothalamus , sleep stages and structure can be objectively determined via comparative analysis of the electroencephalogram (EEG) and electromyogram (EMG) signals. The EEG reflects large scale changes in electrical activity in the cerebral cortex, as the firing rate of cortical neurons steadily declines during non-rapid eye movement (NREM) sleep in comparison to rapid eye movement (REM) sleep and wakefulness –32. AggrThe sleep disorder narcolepsy  highligin vivo use of optogenetics revealed that light-evoked stimulation of Hcrt neurons powerfully promoted wakefulness in mice, an effect dependent on Hcrt (and not other co-released neurotransmitters) signaling  noradrenergic neurons, 5-HT neurons in the dorsal raphe, dopaminergic (DA) neurons within the VTA, histamine neurons in the tuberomammillary nucleus, and cholinergic neurons in the brainstem and basal forebrain . Below, The noradrenergic LC has long been recognized to promote wakefulness and arousal , 55, andThe basal forebrain is a heterogenous structure composed of cholinergic and non-cholinergic (mostly GABAergic) neurons with differential effects on arousal, spanning the substantia innominata, the vertical and horizontal limbs of the diagonal band, the extended amygdala, ventral pallidum, and the medial septum . These aThe TMN contains histaminergic neurons that are silent during sleep and begin to fire after wake onset, where histamine release promotes arousal –77. AcutThe DRN has long been implicated in arousal, showing vigilance state-specific changes in firing rates . ComposeThe laterodorsal tegmental nucleus (LDT) is a key pontine site in the regulation of wakefulness , primariThe VTA is composed primarily of DA and GABAergic neurons and has long been recognized as a node regulating arousal and motivation , 98. EarHcrt peptides can also have direct effects on cortical neurons. These neurons innervate deep layers of the cerebral cortex , and mor+, Ca2+, Mg2+, and H+ was able to cause a reversible switch between sleep and wakefulness. This suggests that downstream of neuromodulator actions , the build-up of certain ionic concentrations in the brain may independently drive sleep pressure and alter the wakefulness threshold.Recently, brain interstitial ion concentrations have been shown to directly influence sleep and wakefulness, providing an indirect pathway by which Hcrt-ergic signaling may influence arousal . NeuromoHcrt neurons respond to a wide variety of neurotransmitters and hormones , includiDo Hcrt receptors (HcrtR1 and HcrtR2) share redundant functions in the regulation of arousal? HcrtR1 binds Hcrt-1 preferentially over Hcrt-2, while HcrtR2 binds both peptides at high affinity . HcrtR2-How do Hcrt neurons integrate time-of-day information to appropriately promote arousal and motivated behavior? The circuitry discussed above largely controls vigilance state switching on short to medium (1–30 s) timescales, so there must be some intervening population of cells that conveys longer term  information to Hcrt neurons to regulate their activity. No direct synaptic connections between the master clock located in the suprachiasmatic nuclei (SCN) and Hcrt neurons has been demonstrated . Howevernptx2b, a gene implicated in AMPA receptor clustering  prevented this response but did not interfere with NMDA-induced Ca2+ increase. The suppressive effects of Hcrt seems to further depend on GABAergic signaling within the SCN, as the inhibitory effect is blocked when Hcrt is applied in the presence of gabazine and CGP55845 (antagonists of GABAA and GABAB receptors). These observations provide evidence for a reciprocal relationship between the SCN and Hcrt neurons in the lateral hypothalamus, demonstrating suppressive and phase-modulatory actions (through Ca2+) of Hcrt on the circadian clock.Hcrt neurons themselves seem to show a circadian rhythm in structural plasticity. In zebrafish, two-photon imaging of Hcrt axons revealed day/night differences in synapse numbers, which was driven by changes in ustering . Overexpustering , suggestustering . By examThe term arousal covers more than mere wakefulness, indeed, it can be considered an umbrella term for multiple waking states, such as restful waking, active waking, and stressed states. Due to the well-studied role for Hcrt initiating arousal from sleep, researchers have also investigated the possibility of a role for Hcrt in the transition from arousal into hyper-arousal associated with stressed states. Supporting evidence for this role has been found in patients suffering from panic symptoms who have elevated levels of Hcrt in their cerebrospinal fluid in comparison to control subjects without substantial panic symptoms . AdditioIn order to further investigate the role of Hcrt in stress researchers have also investigated how manipulation of Hcrt function affects an animal's response to stress. Central administration of Hcrt-1 results in an increase in anxious behaviors as measured in multiple standard laboratory tests of anxiety such as the elevated plus maze, the light-dark box exploration test , the opead libitum-fed controls. Further investigation of interactions between Hcrt, feeding, and stress showed that satiety hormone leptin, which, itself has been shown to be behaviorally anxiolytic  are strongly associated with narcolepsy  challenge reduces cFos expression in Hcrt neurons as well as Hcrt concentrations in cerebrospinal fluid in rats . This isThere are a number of VTA-projecting neuron types in the lateral hypothalamus that are considered to play a role in reward processing. Excitotoxic lesions of the LHA produce similar pathologies to that of dopamine depletion, including aphagia , 170, suHcrt is known to play a role in feeding behaviors. Indeed, studies have shown that Hcrt administration results in increased food intake  and canob/ob and db/db) both strains of mice showed a significant downregulation of the prepro-hypocretin gene  channel (K2P)  . Forty-eP . Fortyel (K2P) , this isel (K2P) . Conversel (K2P) , or had el (K2P) , 190. Thel (K2P)  suggestso leptin –195 a hoo leptin  and is co leptin  and are o leptin . Local Go leptin . Recent o leptin ]. These Hcrt activity is closely linked to not only leptin and insulin, both of which are associated with reducing food intake, but also to the hunger-associated hormone ghrelin which promotes feeding –205. HcrGiven the role Hcrt plays in arousal and motivated states, it is unsurprising that manipulation of Hcrt activity alters many aspects of drug seeking and reward. DiLeone et al.  first suin vivo voltammetry to directly measure dopamine release, España and colleagues demonstrated that Hcrt-1 increases the effects of cocaine on tonic and phasic dopamine signaling, leading to enhanced motivation to self-administer cocaine . Furthermore, they showed that Hcrt fibers innervate the insular cortex, and local blockade of Hcrt signaling in this brain region, but not the adjacent somatosensory cortex, decreased nicotine self-administration . This efWhile studies investigating the function of Hcrt have focused in on a variety of types of arousal such as arousal from sleeping, hyper-arousal in stress-states, arousal from illicit drug-use, etc. there is also a role for Hcrt in behaviors associated with sexual arousal. While the lateral hypothalamus in general has also been associated with inducing  or impaSexual behavior itself can also be modulated via manipulation of Hcrt neurons specifically, by injecting Hcrt-1 into the medial preoptic area significantly improved male sexual behavior by increasing the speed with which males mount females and ejaculate, as well as increasing the frequency with which they mount and ejaculate . ConversHcrt has been shown to play important roles in a wide variety of complex behaviors. The LHA Hcrt system integrates sensory information on stress, metabolic, immune, and motivational states to adjust behavioral responses accordingly. Motivation of behaviors is a two-part process, first comes the energizing component which gives the animal the drive necessary to complete a task, followed by a directional component which determines the type of behavior this burst of energy should be focused toward. The earliest electrophysiological studies in which researchers stimulated neural activity in the LHA showed that activity in this brain region could result in numerous different rewarding behaviors –227, whiResearch on the Hcrt peptides has been unusually productive. Within just 15 years of their discovery, they were causally implicated in the etiology of narcolepsy, specific receptor antagonists were developed, and a dual-receptor antagonist formulation was FDA-approved for the treatment of insomnia. Using cutting edge tools like CRISPR/Cas9 in tandem with cre-dependent adeno-associated viral vectors to manipulate specific genes within Hcrt neurons or their projection sites will undoubtedly provide further classification of their actions and potential for modulation. While a lot of previous work has focused on the role of Hcrt in mediating arousal specifically, the actual operations and computations of the Hcrt network are still unknown. These computations could be achieved by studying projection-selective subsets of Hcrt neurons. It is still unknown whether individual cells perform the same computations, or whether projection specific subsets perform different computations. In order to develop a better understanding of the intricacies of the Hcrt network it will be important for researchers to investigate this neural circuit with all of these systems in mind. While this can appear complicated—it is very likely that the motivated behaviors and the environmental inputs that drive them are all interconnected and can modulate other relevant behaviors to the Hcrt system. Up until this point the heterogeneity of Hcrt neurons specifically, and lateral hypothalamic neurons in general, have slowed the progress of research in this region. With recent developments in research technologies such as optogenetic stimulation, calcium recordings, single cell RNA sequencing, chemogenetic neuron manipulation, and advances in computational modeling, a better understanding of the Hcrt system is within reach. These methods hold much promise for the next 20 years of hypocretin research to continue to be as fruitful as the first twenty.ST and JB wrote the first draft of this manuscript. LdL contributed editorial help and guidance.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.


Previously, a large platinum (Pt) anomaly was reported in the Greenland ice sheet at the Younger Dryas boundary (YDB) . In order to evaluate its geographic extent, fire-assay and inductively coupled plasma mass spectrometry (FA and ICP-MS) elemental analyses were performed on 11 widely separated archaeological bulk sedimentary sequences. We document discovery of a distinct Pt anomaly spread widely across North America and dating to the Younger Dryas (YD) onset. The apparent synchroneity of this widespread YDB Pt anomaly is consistent with Greenland Ice Sheet Project 2 (GISP2) data that indicated atmospheric input of platinum-rich dust. We expect the Pt anomaly to serve as a widely-distributed time marker horizon (datum) for identification and correlation of the onset of the YD climatic episode at 12,800 Cal B.P. This Pt datum will facilitate the dating and correlating of archaeological, paleontological, and paleoenvironmental data between sequences, especially those with limited age control. Petaev et al.18O values, confirming the onset of cooler conditions at the beginning of the YD interval. This peak interval is represented by a rise in Pt concentrations over 14 years and subsequent drop during the following 7 years, consistent with the known residence time of stratospheric dustet al. to have resulted from a highly fractionated, Ir-deficient, iron-rich, extraterrestrial impactor. However, ten additional YDB studies have reported different concentrations and ratios of Pt and other platinum group elements (PGE), including iridium (Ir), osmium (Os), ruthenium (Ru), and rhodium (Rh), all of which usually co-vary  the onset of the YD climate cooling episode at ~12,800 calendar years BP, b) a peak in continental-scale biomass burning, c) extinction of more than 35 genera of North American Pleistocene megafauna, and d) the demise of the Paleoindian Clovis technocomplex345634568910et al.3719et al.et al.A number of other studies have made critical assessments of the evidence for a YD impact event. Specifically, questions have been raised regarding the nature or replicability of the reported impact proxies, the accuracy of the established chronology, and its purported effects on humans and animals at the YD onset121314151617et al.2324et al.et al.et al.et al.Of particular relevance to our study, Andronikov et al.et al.et al.et al.et al.In the same study of Blackwater Draw, Andronikov et al.Andronikov Following the discovery of the Pt anomaly at the onset of the YDB in the Greenland ice sheetWe first tested the hypothesis of a widespread YDB peak in Pt from well-dated, well-stratified sites with clearly established YD-age sediments and with previously reported YDB impact proxies (e.g. microspherules and nanodiamonds). These sites include Arlington Canyon on Santa Rosa Island in Southern California, Murray Springs in southeastern Arizona, Blackwater Draw in eastern New Mexico, and Sheriden Cave in Ohio . We hypoLithologies for western and Midwestern study sites have been well-described elsewhere, but vary widely from cobble lag and alluvial sands and gravels at Arlington Canyon, incised marl deposits filled with stream-channel sands and gravels at Murray Springs, Pleistocene sandy alluvium capped by diatomite and silty muds at Blackwater Draw, and deeply buried, stratified deposits within a collapsed karst cavern at Sheriden Cave  within each sample and found that Pt concentrations may vary widely between duplicate testing of the same aliquot  below the inferred YDB. Although the record at two YDB sites is unclear, 9 out of 11 study sites have Pt anomalies that are constrained to or consistent with the YDB layer based on chronometric dating and/or archaeostratigraphy, thus, making the Pt anomaly a useful time marker horizon. A few other study sites have samples with elevated Pt in post-YDB sediments; however, with only one exception, they represent only slight deviations from background. Although Arlington Canyon has a relatively large Pt anomaly above the YDB, the sampling location is reported to be a catchment basin filled with reworked material, and therefore, would be expected to contain reworked YDB Pt. Multiple and significantly higher Pt anomalies are present within the established YDB interval at Arlington Canyon and are associated with other known impact proxies  is based on numerous studies by multiple authors of artifact distributions, the stratigraphic position of temporally diagnostic artifacts , intact cultural features, occupation surfaces indicated by dense lithic debris, refitting studies indicating minimal vertical displacement, and analysis of sediments indicating discrete periods of sedimentation, artifact burial, and pedogenesis2930313233343530Another anomaly involves the Pt/Pd ratios for the YDB layer, which are typically very different from the background Pt/Pd ratios above and below the YDB layer . Pt/Pd a3456Regarding the likely mechanisms for PGE enrichment, discussed in Sawlowicz34Supplementary Table 10]The platinum anomalies reported here for eastern sites are particularly striking, given the difficulties in establishing archaeological stratigraphy and landform chronologies for the relatively uniform, undifferentiated, sandy sediments that mark the latest Quaternary sequences of the southeastern Atlantic Coastal Plain. Overall, the eastern study sites display few visual changes in stratigraphy that mark the YDB layer, and thus, the Pt anomaly provides a highly useful tool for locating that boundary, particularly because collecting carbon-rich material for radiocarbon dating is very difficult on the Coastal Plain and because OSL dating typically has larger uncertainties at ~13,000 years ago than for younger sediments. In contrast, the YD onset layer is more easily identified at the four intensively studied and well-dated western and Midwestern YDB study sites that have coeval Pt anomalies and YDB proxies, because the anomalies lie directly beneath distinctive, easily identified “black mat” layersRegardless of origin, the consistent presence of anomalous Pt concentrations within sediments from multiple archaeological sites across North America that date to the onset of the YD Chronozone is compelling. This chronostratigraphic marker at the YDB will be useful in geoarchaeological research for inferring the relative stratigraphic position of early Paleoindian (Clovis) and post-Clovis occupations over large portions of North America, especially for sites lacking visual stratigraphy and/or unambiguously stratified, well-dated archaeological assemblages. Regarding limitations of Pt as a chronostratigraphic marker, this study finds that the Pt enrichment is highly variable, may be only slightly above background in some cases, often occurs within narrow (<2.5-cm) sediment intervals, and may be subject to the same post-depositional processes affecting sediments and artifacts .The same limitations discussed above for Pt also apply to methods of absolute dating of archaeological sites. Radiocarbon and OSL dating rely on carbon and sand grains respectively that are subject to reworking and displacement. In the case of the Pt anomalies reported here, their use as a temporal marker for the onset of the YD is likely to be at least as accurate as OSL dating given the large associated error (±500 years or more) typical for OSL dating of 13,000 year-old sediments. Ideally, the Pt anomaly will be useful for identifying areas to be targeted later for absolute dating.et al.This study finds no evidence to contradict the conclusions of Petaev Our evidence indicates the Pt anomaly is a useful time marker horizon, and the ability to locate this chronostratigraphic marker easily within de-glaciated sediments is a significant finding. This time marker will allow more robust age-depth models and facilitate understanding of the effects of YD climate change on early Paleoindians, flora, and fauna across the Bølling-Allerød/YD transition.In all, 199 bulk sediment samples of ~50 gm each were analyzed for Pt and palladium (Pd). A bulk weight of 50 gm or more is strongly recommended due to potential problems from the nugget effect see . Each saHow to cite this article: Moore, C. R. et al. Widespread platinum anomaly documented at the Younger Dryas onset in North American sedimentary sequences. Sci. Rep.7, 44031; doi: 10.1038/srep44031 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Most infections by human papillomaviruses (HPVs) are ‘acute’, that is non-persistent. Yet, for HPVs, as for many other oncoviruses, there is a striking gap between our detailed understanding of chronic infections and our limited data on the early stages of infection. Here we argue that studying HPV acute infections is necessary and timely. Focusing on early interactions will help explain why certain infections are cleared while others become chronic or latent. From a molecular perspective, descriptions of immune effectors and pro-inflammatory pathways during the initial stages of infections have the potential to lead to novel treatments or to improved handling algorithms. From a dynamical perspective, adopting concepts from spatial ecology, such as meta-populations or meta-communities, can help explain why HPV acute infections sometimes last for years. Furthermore, cervical cancer screening and vaccines impose novel iatrogenic pressures on HPVs, implying that anticipating any viral evolutionary response remains essential. Finally, hints at the associations between HPV acute infections and fertility deserve further investigation given their high, worldwide prevalence. Overall, understanding asymptomatic and benign infections may be instrumental in reducing HPV virulence. Alphapapillomavirus genus and classically referred to as ‘high risk types’ [The most oncogenic viruses to humans are a group of around 20, closely related, human papillomavirus (HPV) types. All of them are classified in the k types’ . HPV-indk types’ . Here weInfection-driven cancers are distinctive because they can be fought using the arsenal developed against infectious diseases: identification of risk factors, prevention of transmission and early detection of infected individuals. Identification of risk factors has led to the recognition of a few, closely related oncogenic HPVs as necessary etiologic agents of several cancers . ContagiIn spite of the primary and secondary prevention measurements available, HPVs will continue to infect millions of people in the foreseeable future, thereby causing significant morbidity and mortality worldwide . Indeed,HPV infections that are not chronic or latent have been referred to as ‘acute’ ,19, ‘nonAs illustrated in One important reason for the blurry line between acute and chronic infections is that HPVs are very diverse in terms of genetics as well as in the clinical presentation of the infection. For instance, papillomaviruses colonize the skin and mucosa of virtually all humans from very early in life ,27,28 anFor most viruses the acute, clinically prominent phases of viral infections are generally better understood than the chronic ones . ResearcBesides studies on the natural history of the infection, the placebo arms of vaccine trials have been instrumental in increasing our understanding of the epidemiology of these infections by following thousands of young adults over time e.g., ,36,37). ,37. 36,3Some longitudinal studies performed more frequent sampling over a long period of time. For instance, two studies sampled women every three months. One of these followed unvaccinated girls aged 15–16 years in Tanzania and found that the median time from reported sexual debut to first HPV infection was five months, while median infection duration was six months . AnotherWhy most infections clear, while only few persist and progress to cancer remains largely unresolved . We knoRecent research into the role of innate effectors  and Th-17 responses , we need to invoke the meta-community concept . This haA first advantage of a meta-community approach for HPVs is that it can factor in different target cell types. Papillomaviruses have been infecting amniotes since their first appearance  and haveThe heterogeneous nature of the skin underlies also the different propensity for chronic viral infections to lead to cancer in different anatomical locations. Many squamous cervical carcinomas associated with HPVs are thought to arise from a discrete population of cuboidal epithelial cells located in the transformation zone between the endo- and the ectocervix . The absMore generally, the underlying hypothesis in our meta-population and meta-community approaches is that acute, latent and chronic infections exhibit differential specific features at the virocellular level, which further vary as a function of the infected cell type and differentiation status. Markers of acute versus true persisting infections will most likely be different for different anatomical sites and skin structures. The overall dynamics of the virus–host interaction is thus an integration of the interactions between viral genotype, host genotype, cellular phenotype and environment.Papillomavirus latent infections are still very poorly characterized and understood. The prevalence of latency remains largely unexplored, and its contribution to the oncogenic process in chronic infections is obscure. Establishment of latent infections is a life-history trait shared by very divergent viruses. For large DNA viruses, such as herpesviruses, the infection often starts with an acute phase, and then viral gene expression changes towards a different profile where the virus enters a latent state, with limited or even no genome replication, and no cellular damage, until reactivation is triggered. In small DNA viruses, such as Torque-Teno viruses, the initial infection goes unnoticed  and the viral genomes remain in the host and replicate chronically at very low levels without apparent damage . For papFor HPVs, two ways for latency to arise have been proposed: infections may directly enter latency without going through an acute phase, or it may instead arise after a productive phase without successful clearance . In bothOverall, in addition to molecular virology investigations into latency mechanisms, studies combining a detailed initial follow-up (to demonstrate clearance), a long follow-up (to demonstrate chronic infection) and sequencing  will help generate a complete picture on papillomavirus infection latency. For instance, a recent analysis of HPV16 complete genomes in 57 persistent infections revealed one case of reinfection by the same HPV type, which could have been mistaken for a persitent infection .Since vaccination will not be widespread for many years  and giveE6 and E7 genes of HPV16 and HPV18, which showed efficiency in a controlled trial at improving the regression of high-grade lesions (CIN2/3) [Currently, the bulk of clinical and animal model research into HPV immunotherapies is understandably focused on cervical cancer treatment. Indeed, there are numerous approaches to developing these treatments, such as protein/peptide vaccines, bacteria-and-viral based vectors, and immunomodulators . One of (CIN2/3) . The iat(CIN2/3) ). While In practice, immunotherapies cannot be envisaged to treat all acute HPVs infections given their prevalence and their often sub-clinical presentations. However, knowledge on the acute stage could be transferable to fight chronic stage infections, for instance, by identifying any immune stimulants or immune cell subtypes that help clear natural acute infections. However, it may be possible that several treatments will be needed for different lesion grades , given that the heightened immune suppression microenvironment in advanced malignancies is particularly complex and strong. Therefore, insights from acute infection clearance could be particularly well suited for development of therapies against premalignant lesions that are usually removed surgically.More focused applications could arise from studying HPV infections in sites other than the cervix, for instance in the case of respiratory recurrent papillomatosis. This is a rare condition caused by chronic infection by HPV6 or HPV11 that shares features of both acute and chronic infections. The chronic disease imposes a recurrent burden to the patients to control the acute, benign clinical presentations of the disease, which can progress fatally to the lungs . FurtherThe Zika epidemic has reminded us of the risk viral infections have on pregnancies . AnogeniIn men, anogenital asymptomatic infections by mucosal and cutaneous HPVs are very common  and someFinally, in utero transmission of HPVs is rare , but asyOverall, the presence of mucosal HPVs in placenta and semen suggests non-classical tropisms, and thus, viral life cycle in these particular cellular environments needs to be elucidated. Clinical studies investigating acute anogenital infections in pregnant women  as well as in their partners are required to unravel how HPVs  may be either directly or indirectly increasing the risk of infertility, spontaneous abortions, pre-term labor, pre-eclampsia or other complications.Effective vaccines with high coverage rates exert major selective pressures on pathogens . The risWhile vaccines against HPVs are effective and safe, they may be leaving the door open for an evolutionary response . This cale S5 in ). In theHPV vaccines can generate off-target immune activity and offer protection against closely related viruses not targeted by the vaccine formulation ,97. Yet Thanks to next generation sequencing, we now have access to an increasing number of full genomes. For instance, very recently, an analysis of more than 5500 HPV16 genomes identified shared feature between viruses isolated from precancers and cancers compared to viruses from case-control samples . HoweverEven when viral clearance does occur, recent work shows that acute infections can impair the immune system causing chronic inflammation or ‘immunological scaring’ . CertainSpeculatively, such a long-term impact could concern not only anogenital but also infections at cutaneous sites. Indeed, virtually all humans become infected by very diverse cutaneous HPVs, chiefly beta- and gammapapillomaviruses, which are proposed to act as cofactors for the risk of developing non-melanoma skin cancers in certain human populations . Given tAcute infections by HPVs are the rule rather than the exception, and a better understanding of such infections is urgent because of their enormous fundamental and public health implications. We propose that metapopulation and metacommunity approaches borrowed from ecology can provide a strong explanatory framework to the study of the course of HPV infections and better distinguish between acute, latent and chronic infections. In addition to the promise of identifying early markers of infection chronicity, understanding viral-immunity interactions can help design new treatments to boost natural immunity against novel infections. Such therapies may reduce the chances of an acute infection to chronify and to reach (pre)cancer stages. Detailed studies with long follow-up and short time intervals are further needed to precisely assess the role acute HPV infections could have on fertility or on the long-term ‘immune scars’ these infections may leave behind.Finally, as with most infectious diseases, it is important to remain humble and to accept that elimination  of virulBox 1Do some innate immunity evasion mechanisms lead to delayed clearance? How common is immunological tolerance to HPVs?What role, if any, do natural B-cell responses play in clearance?Are there differences in immunological clearance mechanisms for different HPVs or for different infected cell types, tissue structures or anatomical sites?Are there long term effects after HPV infection clearance ?How do HPV infections affect male and female fertility?How do HPV infection kinetics affect transmission? Are HPVs infections structured in host tissues (the meta-population hypothesis)?What is the role of the microbiota in HPV infection clearance, persistence and progression to cancer? ,114


Epilepsy is a paroxysmal neurological disorder characterized by recurrent and unprovoked seizures affecting approximately 50 million people worldwide. Cognitive dysfunction induced by seizures is a severe comorbidity of epilepsy and epilepsy syndromes and reduces patients’ quality of life. Seizures, along with accompanying histopathological and pathophysiological changes, are associated with cognitive comorbidities. Advances in imaging technology and computing allow anatomical and topological changes in neural networks to be visualized. Anatomical components including the hippocampus, amygdala, cortex, corpus callosum (CC), cerebellum and white matter (WM) are the fundamental components of seizure- and cognition-related topological networks. Damage to these structures and their substructures results in worsening of epilepsy symptoms and cognitive dysfunction. In this review article, we survey structural, network changes and topological alteration in different regions of the brain and in different epilepsy and epileptic syndromes, and discuss what these changes may mean for cognitive outcomes related to these disease states. Epilepsy is a prevalent neurological disorder characterized by recurrent seizures. An important complication of epilepsy and epilepsy syndromes is disruption in cognitive ability. Understanding how cognitive dysfunction occurs will not only improve the quality of life for patients, but will also help elucidate neural mechanisms underlying cognition.There are many forms of epilepsy, broadly characterized as involving partial or generalized seizures. Epilepsy syndromes are defined by associated clinical manifestations. Childhood absence epilepsy (CAE), for instance, is characterized by brief, consciousness-impairing staring spells and 3-Hz discharges on EEG.Some forms of epilepsy and epilepsy syndromes may lead to cognitive impairment, further compounding disease burden. Yet, how cognitive impairment relates to seizures is unclear. How do seizures impact cognition? What are the neural mechanisms mediating this relationship? These are important questions being addressed with investigation on multiple levels.In this review article, we focus on changes to neuroanatomy and neural networks that alter the topological relationship between brain regions. These alterations result in greater seizure susceptibility and cognitive dysfunction. Furthermore, increased seizure severity causes further topological changes, thus establishing a vicious cycle. In this survey, we focus on specific brain areas involved in seizure initiation and cognition and epilepsy syndromes associated with cognitive impairment. Abbreviations used throughout this review article are in Table Animal and clinical studies show that seizures can result in hippocampal dysfunction associated with cognitive and behavioral disturbances in both the developing and mature brain , extra-hippocampal volume abnormalities are observed in association with cognitive dysfunction. Chronic TLE is characterized by extra-hippocampal brain abnormality and cognitive impairment in both memory and non-memory domains , dendritic pathology and cortical alterations.HS is a pathological development associated with benign and drug-resistant mTLE and is involved in epilepsy-related cognitive dysfunction  and the rest of human brain , ELS and juvenile myoclonus epilepsy (JME). In TLE, cortical parameters including area, thickness and volume are affected, and cognitive impairment in TLE is associated with the severity of cortical abnormalities  in children with FLE shows widespread hypometabolism not only in the frontal lobe but also in the extrafrontal cortex, including the temporal and parietal cortices  is an important brain region for neonatal seizure-related cognitive impairment, as in the case of ELS. Anatomical and electrophysiological studies show that direct connections between the hippocampus and medial PFC as part of the hippocampo-PFC circuit play an important role in aspects of learning and memory processing including information consolidation and working memory  alterations in the PFC. STP of the PFC is important to its functions including short-term working memory, information processing and decision-making processes  reveals the following associations between WM in different regions of the brain and cognitive function: (1) right temporal WM with language and executive function; (2) the CC with intelligence and language; and (3) left parietal WM with language , striatal changes in benign epilepsy with centro-temporal spikes (BECTS) and response to hypometabolism in temporal and frontal lobe seizures.When the LSTGp is temporarily impaired by a seizure, brain trauma or stroke, the brain adapts to maintain language comprehension ability. Data from repetitive transcranial magnetic stimulation (rTMS) with fMRI studies suggest that this adaptation consists of two parts: (1) increased synchronization between compensating regions coupled with decreased synchronization within the primary language network; and (2) decreased activation at the rTMS site as well as in distal regions followed by a recovery process. Adaptation involves three synchronization centers: the contralateral homolog of the area receiving rTMS (i. e. the right STGp), areas adjacent to the rTMS site and the medial frontal gyrus, a region involved in discourse monitoring  human epilepsy includes acquired, idiopathic and cryptogenic causes, whereas animal epilepsy models are mostly induced by chemical or physical methods, which means they cannot completely imitate the pathophysiological course of human epilepsy; (2) chemical or physical ignition affects the whole brain and disrupts the neurogenic niche, which makes it difficult to parse out the contribution of specific regions or structures to cognition; and (3) human beings have higher intelligence and more advanced cognitive functions such as computation, language and integrated analysis, which cannot be studied in animal models, thus animal models are used to study memory, social behavior and recognition. For readers interested in chemosonbulsant-induced epilepsy and cognitive testing in animal models, we recommend a review by Minjarez et al. .On the other hand, complicating factors in human studies include the following: (1) most epilepsy patients studied are receiving antiepileptic drugs that can result in cognitive impairment. This acts as a confounding factor when we aim to study the direct relationship between epileptogenesis and epilepsy-associated cognitive deficit; and (2) patients who suffer from chronic epilepsy often have comorbid depression and anxiety, which can result in cognitive decline.Alterations in anatomic structure, networks and topology are associated with seizure-related cognitive impairment. Integrating multiscale imaging approaches such as CT, PET and MRI for macroscopic observation, super-resolution and electron microscopy for microscopic research, and computational tools for analysis, studying structural or sub-structural alterations in the brain is a promising and rewarding approach to understand brain function and dysfunction.QW: conception and design, manuscript writing, final approval of manuscript. CWZ and ZL: conception and design, final approval of manuscript. BX: final approval of manuscript. LF: conception and design, financial support, final approval of 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. The reviewer JIA declared a shared affiliation, though no other collaboration, with two of the authors CWZ and LF to the handling Editor.


Among the complex determinants of aging, mitochondrial dysfunction has been in the spotlight for a long time. As the hub for many cellular functions, the maintenance of an adequate pool of functional mitochondria is crucial for tissue homeostasis. Their unique role in energy supply makes these organelles essential, especially in those tissues strictly dependent on oxidative metabolism. Mitochondrial quality control (MQC) is ensured by pathways related to protein folding and degradation as well as by processes involving the entire organelle, such as biogenesis, dynamics, and mitophagy. Dysfunctional MQC, oxidative stress and inflammation are hallmarks of senescence and chronic degenerative diseases. One of the consequences of age-related failing MQC and oxidative stress is the release of mitochondria-derived damage-associated molecular patterns (DAMPs). Through their bacterial ancestry, these molecules contribute to mounting an inflammatory response by interacting with receptors similar to those involved in pathogen-associated responses. Mitochondrial DAMPs, especially cell-free mitochondrial DNA, have recently become the subject of intensive research because of their possible involvement in conditions associated with inflammation, such as aging and degenerative diseases. Here, we review the contribution of mitochondrial DAMPs to inflammation and discuss some of the mechanisms at the basis of their generation. Aging is a complex and multi-factorial process characterized by increased risk of adverse health outcomes . Understmilieu that characterizes aging and its associated conditions are not completely understood.Together with mitochondrial dysfunction, chronic inflammation is another hallmark of both aging and degenerative diseases . InteresHere, we provide an overview of major processes of MQC and their changes during aging. Subsequently, we describe candidate mechanisms responsible for generating and releasing mitochondrial DAMPs. Finally, we summarize the current evidence in support of mitochondrial DAMPs as triggers for age-related chronic inflammation.The mitochondrial proteolytic quality control system consists of subcompartment-specific proteases (mitoproteases) and the ubiquitin-proteasome system (UPS) that together regulate mitochondrial protein turnover and degrade misfolded or oxidized proteins . MitochoMitochondrial protein turnover is also regulated by the cytosolic UPS . A proteSimilar to the endoplasmic reticulum (ER), mitochondria possess a stress responsive system for protein degradation that shares with its ER analogue some key components, including the AAA ATPase p97 and the cofactor Npl4 . Under sMitochondrial biogenesis is a multi-stage process generating new organelles that, in a dynamic balance with their degradation, regulate the mitochondrial content within the cell. Mitochondriogenesis is orchestrated through the expression of nuclear and mtDNA-encoded genes. In particular, transcriptional coactivators belonging to the peroxisome proliferator activated receptor gamma coactivator-1 (PGC-1) family , the nuclear respiratory factor (NRF) 1 and 2, and the estrogen-related receptor α (ERRα), coordinate the expression of mitochondrial proteins encoded by nuclear DNA . SubsequBoth qualitative and quantitative changes in mtDNA have been found with aging in various tissues and across species ,28. MtDNThe importance of the modulation of TFAM binding also resides in its ability to stabilize the mtDNA. The latter, if unbound, becomes more fragile and prone to degradation, which ultimately leads to organelle dysfunction. The identification of misplaced TFAM and mtDNA in the plasma of persons with chronic inflammatory diseases suggests these molecules may be involved in the inflammatory response ,38. WhetMitochondrial dynamics control organelle shaping, metabolic plasticity , redox sThe integration of mitochondrial dynamics and mitophagy ensures an efficient MQC process and contributes to preserving metabolic cellular “fitness” and plasticity . DerangeThe exposure of cultured cells to subcytotoxic doses of hydrogen peroxide represses the expression of fission protein 1 (Fis1), thereby promoting the formation of elongated mitochondria with increased oxidant emission . ExcessiImbalanced mitochondrial dynamics toward fission have been found in several disease conditions  as well Autophagy is, literally, a cellular self-eating process through which intracellular components are degraded within lysosomes during periods of stress, such as nutrient deprivation, as an attempt to adapt and survive . SelectiDysfunctional vesicle trafficking as well as specific molecular patterns recruited in vesicle cargos has been described under several conditions ,62. A seInflammation has long been considered to be a defense response against microbial agents. It is now clear that a (chronic) inflammatory response can also occur in the absence of infections, a condition referred to as “sterile inflammation” . The terThe anti-microbial inflammatory response and sterile inflammation are alternatively mounted through the recruitment of distinct macrophage subsets: (1) tissue-resident macrophages, which serve anti-inflammatory functions to safeguard tissue homeostasis and resolve local inflammation; and (2) circulating monocytes, which are pro-inflammatory and limit the spread of the infection .Mitochondria play a central role in sterile inflammation through the activation of several pathways . A redoxParticularly interesting are the differential outcomes of redox imbalance in inflammation. In the setting of moderate inflammation, if cellular repair systems are overwhelmed, the intrinsic apoptotic cascade may be triggered . In the Another pathway through which mitochondria contribute to sterile inflammation is deeply rooted into the “danger theory” of inflammation proposed by Matzinger . This paThe TLR pathway is triggered by the binding of DAMPs to neutrophils, followed by their activation and subsequent organization of the inflammatory response via NF-κB signaling  Figure . An alteThe cGAS-STING DNA-sensing pathway is an additional component of the innate immune system . Upon biBeside a generalized pro-oxidant environment, aging is also characterized by a decline in immune cell subtypes and functions, a condition referred to as immunosenescence . Aged imEver since Matzinger proposed the “danger theory” of inflammation , severalInterestingly, circulating levels of mtDNA molecules increase progressively past the age of 50 and correlate with those of pro-inflammatory cytokines, including IL6, tumor necrosis factor α (TNF-α), Regulated on Activation Normal T Cell Expressed and Secreted (RANTES), and IL1 receptor antagonist . RemarkaTFAM has also been suggested to act as a mitochondrial DAMP . Mouse eGeneral autophagy and mitophagy modulate inflammation by either clearing apoptotic corpses through macrophage activity or inhibiting NLRP3 inflammasome activation. Down-regulation of mitophagy results in spontaneous inflammasome activation as a consequence of mitochondrial ROS burst ,92. Accomilieu of aging represents an uninvestigated but interesting scenario.Cell death has been suggested to be another modulator of immune response through mitochondrial involvement and inflammasome activation . ATP relDespite the evidence supporting cell-free mtDNA and TFAM-bound mtDNA as DAMPs, the exact mechanisms of mtDNA delivery into the cytosol and then into the circulation are currently unknown. As elegantly reviewed by Safdar et al. , one of milieu have been proposed to contribute to inflamm-aging. From this perspective, aging may be envisioned as the result of an “autoimmune-like” process. Given the role played by mitochondrial DAMPs in the activation of sterile inflammation, the mechanisms favoring organelle damage, in particular failing MQC processes, represent a relevant matter to be addressed by future investigations. The elucidation of these mechanisms may provide clinicians with novel therapeutics to counteract inflamm-aging and its negative correlates.Population aging poses a tremendous burden on the society. This has instigated intense research on the mechanisms that make the elderly more susceptible to diseases and disability. Several processes have been identified. Among these, inflamm-aging, a condition of chronic inflammation that develops independent of infections, has gained special attention. The cellular mechanisms responsible for inflamm-aging are not fully understood. However, recent studies suggest that a danger cellular-driven response may represent a relevant player. The coexistence of oxidative stress resulting from mitochondrial dysfunction and sterile inflammation has been summarized in the concept of oxy-inflamm-aging that merges the role of inflammation and oxidative stress in the aging process. Specific “danger molecules” generated in an oxidative