Growing evidence from our group and others indicates that key neural functions, including regulation of synaptic plasticity and axonal guidance and connectivity, arise from interactions between glial cells, neurons, and the extracellular matrix. Several distinct populations of glial cells critically contribute to the composition of main components of the extracellular matrix (ECM), synthesizing them and secreting them into the extracellular space, where they become incorporated in organized ECM structures. The brain ECM, and chondroitin sulfate proteoglycans (CSPGs) in particular, play a key role in brain development and adult life, in turn regulating glial functions as well as synaptic plasticity and neural connectivity. We have previously shown that glial cells expressing CSPGs are altered in the amygdala and entorhinal cortex of people with schizophrenia (SZ) and bipolar disorder (BD). These changes are accompanied by marked decreases of perineuronal nets (PNNs), organized ECM structures unsheathing distinct neuronal populations. Recent and ongoing studies are focused on novel CSPG-enriched ECM structures, related to synaptic complexes and myelinated axons, their relationship to glial populations and their involvement in the pathophysiology of SZ and BD.Postmortem tissue samples from the amygdala, entorhinal cortex and thalamus from a well characterized cohort of healthy control, SZ and BD subjects were included in these studies. Multiplex immunofluorescence combined with quantitative microscopy was used to quantify glial cells and CSPGs, while electron microscopy on human and mouse tissue were used to investigate ultrastructural morphology. Step-wise ANOVA analyses included several potential confounds such as exposure to pharmacological agents and substance abuse.Our results show that at least two novel ECM structures are present in the human brain. The first, enriched in CSPGs bearing chondroitin sulfation in position 6 (CS-6), and named here ‘CS-6 clusters’ was found to be markedly decreased in the amygdala of people with SZ and BD. Electron microscopy studies show that CS-6 clusters are composed of astrocytes synthesizing and secreting CS-6 CSPGs in the vicinity of adjacent groups of dendrites, where it is incorporated into postsynaptic densities of dendritic spines. The second CSPG-enriched ECM structure, i.e. axonal coats, has been observed in the human thalamus to envelope distinct populations of axons, interweaving with myelin sheets. Its main CSPG components appear to be synthesized and secreted by oligodendrocytes precursor cells located in the vicinity of axon bundles. Preliminary results show abnormalities affecting both oligodendrocyte precursors and axonal coats in SZ.In summary, our results show complex interactions between glial cells, neurons and ECM, potentially affecting synaptic functions and axonal conductance. Results in SZ and BD point to a profound disruption of these interactions in several brain regions.


Diabetes mellitus in early pregnancy is the most severe maternal disease that is counted for 10% of newborn infants with structural defects. With the rapid increases in the number of diabetic women in childbearing age, the birth defect rate is projected to elevate dramatically. Thus, prevention of embryonic malformations becomes an urgent task. Animal studies have revealed an involvement of oxidative stress in diabetic embryopathy and treatment with antioxidants can reduce embryonic abnormalities. However, the failure of clinical trials using free radical-scavenging antioxidants to alleviate oxidative stress-related diseases prompts researchers to reevaluate the strategy in birth defect prevention. Hyperglycemia also disturbs other intracellular homeostasis, generating aberrant conditions. Perturbed folding of newly synthesized proteins causes accumulation of unfolded and misfolded proteins in the lumen of the endoplasmic reticulum (ER). The ER under the stress activates signaling cascades, known as unfolded protein response, to suppress cell mitosis and/or trigger apoptosis. ER stress can be ameliorated by chemical chaperones, which promote protein folding. Hyperglycemia also stimulates the expression of nitric oxide (NO) synthase 2 (NOS2) to produce high levels of NO and reactive nitrogen species and augment protein nitrosylation and nitration, resulting in nitrosative stress. Inhibition of NOS2 using inhibitors has been demonstrated to reduce embryonic malformations in diabetic animals. Therefore, targeting ER and nitrosative stress conditions using specific agents to prevent birth defects in diabetic pregnancies warrant further investigations. Simultaneously targeting multiple stress conditions using combined agents is a potentially effective and feasible approach. Congenital fetal anomalies are a major factor in perinatal mortality and infant disability. According to the World Health Organization, approximately two hundred seventy thousand newborn babies die each year from congenital abnormalities . In the Diabetes mellitus in early pregnancies is the most severe maternal disease to cause birth defects in newborn infants, in addition to other adverse pregnant outcomes including small for gestational age (growth restriction), large for gestational age (macrosomia), and perinatal demise  –8. AlthoIt has been shown that maternal hyperglycemia alters mitochondrial morphology and function, leading to generate high levels of reactive oxygen species (ROS) ,12. ConcThe identification of oxidative stress in diabetic embryopathy led to development of strategies to reduce fetal abnormalities using antioxidants in animal models . Lipoic Increasing the expression of endogenous antioxdative enzymes may enhance the intracellular defense against hyperglycemic insult. It has been shown that, in strains of rat that express higher levels of SOD and catalase, embryos of diabetic dams are resistant to hyperglycemic insult . In diabAlthough animal studies have shown that antioxidants can reduce developmental malformations in embryos, the enthusiasm about the application to humans has been dampened by the failure in multiple large scale trials using antioxidants  to treat similar diseases  –41. The Indeed, stress in the endoplasmic reticulum (ER), exhibited by changes in specific factors (markers), in the embryos of diabetic animals has been observed –45. HypeER stress can be alleviated by enhancing protein folding, using chemical chaperones, including phenylbutyrate (PBA), ursodeoxycholic acid (UDCA), and taurine-conjugated derivative, tauroursodeoxycholic acid (TUDCA) . The cheIn addition to dysfunction of the ER, maternal hyperglycemia also stimulates production of nitric oxide (NO) in the embryo, notably in the central nervous and cardiovascular systems ,56. NO iNO is produced by members of the NO synthase (NOS) family, consisting of NOS1 , NOS2 (inducible NOS or iNOS), and NOS3  –66. nNOSIn the embryos of diabetic animals, NOS2 expression is increased, whereas the expression of NOS1 and NOS3 is decreased ,73. The The failure of antioxidant trials apparently casts clouds over the effort to apply the similar approach to birth defect prevention; but it also opens opportunities for developing new strategies. Instead of scavenging free radicals, reinforcing the endogenous antioxidative capacity is an approach potentially to achieve effectiveness. This approach includes upregulation of antioxidative enzymes and replenishing of endogenous antioxidants.NAC, a cysteine precursor, and folic acid (FA), a methionine precursor, can increase biosynthesis of GSH to enhance intracellular protection against ROS ,75. TreaIn addition to the strategy of replenishing endogenous antioxidants, upregulation of antioxidative enzymes is also a powerful approach. This notion is supported by the experiments, in which overexpression of a superoxide dismutase in mouse embryos can decrease malformation rate in diabetic pregnancies ,82. Animin vivo . Each of the conditions can be intervened with pharmacological approaches in vivo . Targeti


In the setting of profound ocular blindness, numerous lines of evidence demonstrate the existence of dramatic anatomical and functional changes within the brain. However, previous studies based on a variety of distinct measures have often provided inconsistent findings. To help reconcile this issue, we used a multimodal magnetic resonance (MR)-based imaging approach to provide complementary structural and functional information regarding this neuroplastic reorganization. This included gray matter structural morphometry, high angular resolution diffusion imaging (HARDI) of white matter connectivity and integrity, and resting state functional connectivity MRI (rsfcMRI) analysis. When comparing the brains of early blind individuals to sighted controls, we found evidence of co-occurring decreases in cortical volume and cortical thickness within visual processing areas of the occipital and temporal cortices respectively. Increases in cortical volume in the early blind were evident within regions of parietal cortex. Investigating white matter connections using HARDI revealed patterns of increased and decreased connectivity when comparing both groups. In the blind, increased white matter connectivity (indexed by increased fiber number) was predominantly left-lateralized, including between frontal and temporal areas implicated with language processing. Decreases in structural connectivity were evident involving frontal and somatosensory regions as well as between occipital and cingulate cortices. Differences in white matter integrity  were also in general agreement with observed pattern changes in the number of white matter fibers. Analysis of resting state sequences showed evidence of both increased and decreased functional connectivity in the blind compared to sighted controls. Specifically, increased connectivity was evident between temporal and inferior frontal areas. Decreases in functional connectivity were observed between occipital and frontal and somatosensory-motor areas and between temporal (mainly fusiform and parahippocampus) and parietal, frontal, and other temporal areas. Correlations in white matter connectivity and functional connectivity observed between early blind and sighted controls showed an overall high degree of association. However, comparing the relative changes in white matter and functional connectivity between early blind and sighted controls did not show a significant correlation. In summary, these findings provide complimentary evidence, as well as highlight potential contradictions, regarding the nature of regional and large scale neuroplastic reorganization resulting from early onset blindness. Ocular blindness has served as an important model in helping to understand the consequences of sensory deprivation on brain development. Extensive work in animal models has provided compelling anatomical and behavioral evidence regarding the dramatic neuroplastic changes that result from altering visual experience e.g. –3). In h. In h3])Despite accumulating evidence of these dramatic neuroplastic changes , a number of reports have suggested that the functional organization (i.e. domain specificity) of the occipital cortex may develop independently of visual experience e.g. –31). Imp. Imp31])At the regional level, structural morphometry studies of the occipital cortex in blind humans show evidence of decreased gray matter volume , 34–36 aCertainly, these results continue to raise interesting questions regarding the nature and extent of neuroplastic changes within the context of profound blindness. However, uncertainty remains regarding the relationship between changes in brain structure and large-scale changes in anatomical and functional brain connectivity. This is particularly evident when one considers that previous imaging studies have usually focused on analyzing one parameter at a time, thus making reconciliation of observed results across multiple studies particularly challenging. Given these gaps in our understanding, it would be of value to capture multiple measures of morphometry, structural, and functional data throughout the brain as a means to better characterize the nature and extent of these changes resulting from visual sensory deprivation.To our knowledge, no previous study has attempted to address this issue using a comprehensive multimodal imaging approach. Here, we investigated potential differences between early blind and sighted control individuals with respect to morphometry obtained from standard anatomical MRI, white matter connectivity and integrity obtained by diffusion MRI, as well as functional connectivity characterized by resting state fMRI. In so doing, we aimed to examine the interrelationship between these multiple imaging parameters, with particular attention to structural and functional connectivity.A total of 28 subjects were recruited for the study and separated into two groups comprised of 12 early blind  and 16 normally sighted controls . Comparing demographic factors between both groups revealed no statistically significant differences in terms of age (p = 0.26) or gender (Chi-square = 0.86). For the purposes of this study, we defined “early blind” as documented residual vision no greater than light perception and/or hand motion acquired prior to the age of three . While 1-weighted scans were acquired using a turbo spin echo sequence . Diffusion based imaging was carried out using high angular resolution diffusion imaging (HARDI). Images were acquired with a single-shot EPI sequence . Resting state functional connectivity (rsfcMRI) was acquired with a 7 min single-shot EPI sequence . For the resting state sequence, subjects were blindfolded and instructed to lie still and let their minds “wander” while remaining awake. Finally, a field map was acquired to correct for EPI related field inhomogeneities using a fast field echo sequence .All imaging data were acquired on a 3T Philips Achieva System  with an 8-channel phased array coil. Subjects were instructed to lie still and foam padding was used to minimize head motion. Two structural TData processing procedures and analysis workflow for structural morphometry, HARDI-based tractography, and rsfcfMRI analyses are outlined in 1-weighted scans were processed using FreeSurfer 5.3.0 (https://surfer/nMR/mgh/harvard.edu). Details regarding this procedure have been described elsewhere ).To assess differences in connectivity (HARDI and rsfcMRI) between individuals in the sighted and blind groups, a two-sample t-test was performed for each connection between pairs of ROIs. This resulted in a total of 2278 (i.e. 68 X 67/2 = 2278) connections. A False Discovery Rate (FDR) analysis was performed at a rate of q = 0.05 to correct for multiple comparisons within the structural and functional connectivity matrices  as previAs a final analysis, a Pearson’s r-coefficient was used to determine the degree of association between the acquired measures of structural (number of fibers obtained by HARDI) and functional  connectivity. Prior to correlation analyses, HARDI matrices were z-transformed to ensure normality. Correlations between early blind and sighted control subjects for both white matter fiber number and resting state connectivity were examined separately. Finally, to explore if alterations in the profile of white matter connectivity were related to alterations in the profile of resting state functional connectivity, correlations were determined between changes in structural connectivity (i.e. early blind and sighted controls) with changes in functional connectivity (i.e. early blind and sighted controls). For this correlation analysis, the difference between groups was calculated by subtracting the mean sighted control connectivity data from the mean ocular blind connectivity data (separately for HARDI and for rsfcMRI data). These difference matrices were z-transformed prior to the final correlation analysis.http://www.circos.ca/) and processing pipelines adapted for neuroimaging data have been previously explained in detail [The term connectome has been used to refer to the development of comprehensive maps characterizing neural connections within the brain . In thisn detail . Brieflyn detail ;).An initial exploratory analysis  revealed widespread differences (i.e. trends of both increases and decreases) in cortical volume throughout both hemispheres when comparing early blind to sighted controls. Specifically, clusters showing increases in cortical volume were observed bilaterally in the temporal, frontal, cingulate, and motor  cortices, as well as the right parietal cortex. Decreases in cortical volume in early blind were more widespread and clusters showing decreases in volume were observed bilaterally in the occipital, temporal, parietal, and frontal, as well as the left cingulate and sensorimotor  cortices (p<0.005 uncorrected). In the subsequent analysis, only two of these clusters survived correction for multiple comparisons. Specifically, a significant decrease in volume of the left pericalcarine cortex (p = 0.0066) and a significant increase in volume of the right inferior parietal cortex (p = 0.0032) was evident when comparing early blind to sighted controls . However, the only cluster to survive correction for multiple comparisons was a significant decrease observed in the left fusiform gyrus .Exploratory analysis  of changes in white matter volume revealed that early blind subjects showed decreases compared to sighted controls within occipital regions including bilateral pericalcarine, cuneus, and lingual cortices, as well as the right lateral occipital region. There was no significant trend for relative increases in white matter volume in blind subjects compared to controls.Finally, comparing subcortical structures  did not reveal statistically significant differences between early blind and sighted controls . Complete results from the morphometric analyses (uncorrected) are reported in Exploratory analysis of ROI-pairs revealed a number of connections that differed significantly in early blind subjects compared to sighted controls (p<0.05). Specifically, increased connectivity (indexed by fiber number) in early blind compared to sighted was observed for bilateral intra-hemispheric and inter-hemispheric connections involving temporal, parietal, and frontal lobes, as well as the left occipital and sensorimotor cortices . Five ofAn analysis of QA was also carried out as an index of white matter integrity. Exploratory analysis of ROI-pairs revealed a number of connections that differed for QA in early blind subjects compared to sighted controls (p<0.05). Specifically, increased QA was observed in early blind compared to sighted for bilateral intra-hemispheric connections involving the occipital, temporal, parietal, frontal, and cingulate cortices. Additional inter-hemispheric increases in QA were also observed, however these were less numerous than intra-hemispheric increases. Increased QA was observed in five connections following FDR correction for multiple comparisons. Increases in QA in the early blind compared to sighted were mainly left lateralized involving fronto-temporal and temporal-temporal white matter connections. Exploratory analysis also revealed a number of connections showing decreases in QA in early blind compared to sighted subjects (p<0.05). These were equally distributed between inter-and intra-hemispheric connections and involved occipital, temporal, parietal, frontal, and cingulate cortices. Decreased QA was observed in 14 connections after FDR correction for multiple comparisons. Decreases in QA included the interhemispheric connections between the occipital lobes through the splenium were among those that survived FDR correction. Further decreases surviving FDR correction included the left occipito-cingulate, left entorhinal to insula, as well as right lingual to parahippocampal gyrus, and right entorhinal to precentral gyrus . Notably, increases in temporal correlations in early blind compared to sighted involved bilateral intra-hemispheric occipital, temporal, and frontal connections, as well as inter-hemispheric connections involving the occipital, temporal, frontal, and sensori-motor cortices . Of the In the exploratory analysis, there was an overall greater trend for decreases rather than increases in temporal correlations between regions when comparing early blind to sighted. In particular, bilateral intra-hemispheric and inter-hemispheric connections involving the occipital, temporal, frontal, parietal, cingulate, and sensorimotor cortices . Of the As a final analysis, the degree of association characterizing structural connectivity (i.e. white mater fiber number obtained by HARDI) and functional connectivity  between early blind and sighted controls was independently determined. Regarding white matter structural connectivity, the measure of association was found to be very high  .To help better visualize the inter relationships between all the data modalities acquired, the results of the exploratory morphometry analysis , white matter connectivity (uncorrected), and functional connectivity (uncorrected) were presented simultaneously using a circular connectogram , occipital cortical regions , and regions involved in higher order cognitive functions .In summary, outcomes of morphometry in early blind compared to sighted controls revealed co-occurring decreases in cortical volume and cortical thickness within visual processing areas of the occipital and temporal cortices respectively. Increases in cortical volume in the early blind were evident within regions of parietal cortex (corrected results). White matter connectivity in the blind was increased predominantly within the left hemisphere, including between frontal and temporal areas implicated with language processing. Decreases in structural connectivity were evident involving frontal and somatosensory regions as well as between occipital and cingulate cortices. Changes in white matter integrity (as indexed by QA) were also in general agreement with observed pattern changes in the number of white matter fibers. Analysis of resting state sequences revealed patterns of increased connectivity between temporal and inferior frontal areas in the blind while decreases in functional connectivity were observed between occipital and frontal and somatosensory-motor areas and between temporal (mainly fusiform and parahippocampus) and parietal, frontal, and other temporal areas. Finally, correlations in white matter connectivity and functional connectivity observed between early blind and sighted controls showed an overall high degree of association. However, comparing the relative changes in white matter and functional connectivity between early blind and sighted controls did not show a significant correlation.While extensive differences were observed along all three MRI-based modalities, the most notable finding was the evidence of increased inter- and intrahemispheric white matter connectivity in blind individuals. Taken together, the combination of these morphological, structural, and functional connectivity changes may underlie the neurophysiological substrate that supports crossmodal sensory processing and compensatory behaviors observed in individuals who are blind.Morphological changes in gray and white matter as well as subcortical structures have been investigated previously by a number of groups. In general, our differences in morphological findings from the exploratory analysis are in agreement with previous studies using both volume and surface based morphometry approaches investigating cortical and subcortical morphometry. We observed trends for co-occurring decreased cortical volume and increased cortical thickness within occipital cortex in the blind as compared to sighted controls as reported by other groups –40, 80. Notably, we did not find statistically significant differences between both groups with regards to the morphometry of subcortical structures  although a previous group of studies did report increased hippocampal volume in the blind , 83, 84.In contrast to previous reports, we found evidence of both significant increases and decreases in white matter connectivity (both inter- and intrahemispheric) in the blind compared to sighted controls. The vast majority of prior studies have highlighted evidence of marked reductions in the structural volume of numerous brain areas, and in particular, geniculocalcarine structures and tracts including the optic radiations , 42, 44.As opposed to these aforementioned studies employing DTI, our study concentrated on white matter whole-brain connectivity patterns reconstructed by diffusion data acquired by HARDI. Using fiber number as the main outcome of interest, we found evidence of both increases and decreases in white matter connectivity in the blind compared to sighted controls. These observed white matter changes were also supported by analyzing an index of white matter structural integrity: quantitative anisotropy (QA). In particular, we found evidence of increased connections involving superior temporal cortex; a region of the brain that is involved in spatial awareness .The reason for our ability to observe both significant increases and decreases in white matter connectivity (as opposed to only decreases as in previous studies) can likely be explained by the variant of the diffusion MRI approach used in this study. In contrast to previous studies carried out with blind individuals, we employed HARDI rather than DTI to characterize white matter connectivity and integrity. While both techniques provide information regarding the degree of water diffusivity in the brain in order to derive local axonal fiber orientation, DTI is unable to correctly resolve multiple individual orientations within the same voxel such as crossing fibers . In conFinally, with regards to resting state functional connectivity, our results are in general agreement with previous findings reporting patterns of both increases and decreases in connectivity in the blind. In a recent study, Burton and colleagues (2014) found evidence of decreased functional connectivity between occipital cortex and auditory as well as somatosensory cortices in early blind individuals as compared to normally sighted controls. In contrast, increased functional connectivity was reported between occipital and frontal cortical areas  along with parietal regions of the brain . EnhanceAs mentioned earlier, there has been considerable interest in relating compensatory behaviors in blind individuals within the context of observed structural and functional neuroplastic changes in the brain. While compensatory abilities in the blind have been reported across a wide variety of behavioral tasks and implicating different sensory modalities, it is important to note that these abilities are not universally evident across the blind population. Indeed, while there is evidence of enhanced sensory and cognitive task performance, there are also other reports suggesting that the blind are equal or even impaired on certain tasks compared to the their sighted counterparts , 13, 95.In this study, we acquired spatially co-registered data from three MRI-based modalities; structural morphometry, HARDI, and resting state. When comparing across modalities, it is perhaps surprising to find evidence of inconsistencies between increased white matter connections and decreased functional connectivity in early blind and sighted individuals. In other words, while we observed increases in white matter connectivity in the blind, these connections were not always associated with concomitant enhancement in functional connectivity.The reason for this apparent mismatch is not entirely clear at this time. However, such incongruities in observed structural and functional connectivity have been previously noted. A classic example is the evidence of strong interhemispheric functional connectivity between primary visual cortices despite a lack of direct white matter links between these same two regions . While tRecent work has also highlighted important differences in functional connectivity derived from resting state sequences with that of functional connectivity that characterizes task-specific activity. More specifically, it has been suggested in a number of reports that differences in connectivity patterns between populations are task dependent –103 and To help resolve these inconsistencies, further studies should compare metrics of brain structure, white matter structural connectivity, along with measures of effective connectivity as well as individual parametric measures of behavioral performance. Again, by leveraging the advantages of multi-modal imaging, we are more likely to better understand how structure, connectivity, and behavior are reciprocally linked.S1 FigSurface based morphometry analysis comparing differences in A) cortical volume and B) cortical thickness between early blind and sighted controls (corrected for multiple comparisons). Decreases in cortical volume (shown in blue) were evident within the left pericalcalcarine region, while increases (shown in red) were evident in right inferior parietal cortex. Decreases in cortical thickness were observed within left fusiform cortex (shown in blue). Differences in white matter volume and subcortical structures were not significant following correction for multiple comparisons. Abbreviations: L = left, R = right.(EPS)Click here for additional data file.S2 FigQuantitative anisotropy (QA) values revealed by HARDI. A) Exploratory analysis of ROI-pairs revealed an overall tendency for both increases and decreases in QA in ocular blind compared to sighted controls. Decreases were spread throughout both inter- and intra-hemispheric connections, whereas increases in QA were primarily limited to inter-hemispheric connections. B) Ball and stick representation of increases (upper panel) and decreases (lower panel) in QA values between early blind and sighted controls after FDR correction. Differences in connection strength are represented by line thickness, whereby thicker lines represent larger changes in the early blind group . Increases in QA in the early blind group are represented by orange lines, whereas decreases in QA in the ocular blind are represented by cyan lines. Dark blue spheres represent the nodes (i.e. ROIs) associated with the start and/or end points of the connections. Note the overall similarity to the pattern presented for white matter connectivity (fiber number) shown in (EPS)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.


Over-production of reactive oxygen species (ROS) in plants under stress conditions is a common phenomenon. Plants tend to counter this problem through their ability to synthesize ROS neutralizing substances including non-enzymatic and enzymatic antioxidants. In this context, ascorbic acid (AsA) is one of the universal non-enzymatic antioxidants having substantial potential of not only scavenging ROS, but also modulating a number of fundamental functions in plants both under stress and non-stress conditions. In the present review, the role of AsA, its biosynthesis, and cross-talk with different hormones have been discussed comprehensively. Furthermore, the possible involvement of AsA-hormone crosstalk in the regulation of several key physiological and biochemical processes like seed germination, photosynthesis, floral induction, fruit expansion, ROS regulation and senescence has also been described. A simplified and schematic AsA biosynthetic pathway has been drawn, which reflects key intermediates involved therein. This could pave the way for future research to elucidate the modulation of plant AsA biosynthesis and subsequent responses to environmental stresses. Apart from discussing the role of different ascorbate peroxidase isoforms, the comparative role of two key enzymes, ascorbate peroxidase (APX) and ascorbate oxidase (AO) involved in AsA metabolism in plant cell apoplast is also discussed particularly focusing on oxidative stress perception and amplification. Limited progress has been made so far in terms of developing transgenics which could over-produce AsA. The prospects of generation of transgenics overexpressing AsA related genes and exogenous application of AsA have been discussed at length in the review. Likewise, ascorbate anion (AH−) is a vital water-soluble antioxidant molecule within biological system ] is an antioxidant molecule and a key substrate for the detoxification of reactive oxygen entities  improved chlorophyll “a” contents of wheat seedlings subected to salt stress. Similarly, an increase in plant growth and proline content and decrease in ion leakage and lipid peroxidation was observed in okra plants under drought stress  via seed soaking, rooting medium and foliar spray on wheat plants under salinized hydroponic culture. They found that AsA application improves growth, AsA content, and activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) antioxidative enzymes.Not only this, AsA is a ubiquitious molecule proves effective in improving stress tolerance in plants Table . Most efGenetic engineering to improve abiotic stress tolerance is one of the important strategies being under consideration these days. A variety of genetically engineered crop plants have been generated particularly with superior salinity and drought tolerance. Potentially, AsA can improve plant growth and yield as a potential regulator of different mechanisms under adverse factors, so over-accumulation of AsA in plants through gene (s) engineering could efficiently increase plant stress tolerance. Yet, not a single report in the literature is available on stress tolerant genetically modified plants with high accumulation of AsA. Thus, under stress conditions the genetic manipulation of plants with the goal of obtaining high accumulation of AsA is an essential area to be considered. Thus, the present review focuses on how biosynthesis of AsA is regulated in plants under stress or non-stress conditions and how far AsA accumulation in plants has been improved by different means.primary most elucidated pathway is the Smirnoff-Wheeler pathway which is also called as D-mannose/L-galactose pathway  than catalase (CAT) and peroxidase (POD) and the exogenous addition of ascorbate through the nutritional solution was reported to result in marked reduction in the activities of both enzymes  system is far efficient in detoxification of hydrogen peroxide  is reported to be localized in the cytoplasm, however, in apoplast its concentration is reported in milli-molar range due to its export from the cytosol  is a glycoprotein and a member of blue copper oxidase family that mediates ascorbate oxidation to dehydroascorbate (DHA) which usually follows high affinity transport across the membrane  the involvement of DHA in signaling events, and (4) possible involvement of (APX) in the apoplast.Interestingly, the function of this enzyme in the past was often confused or neglected in several studies, so it requires to be elucidated well. The extracellular localization of this enzyme proposes its role as a link between environmental stress perception and adaptive response. Recently, in this connection, De Tullio et al.  proposedStriking functional similarities exists for (APX) and (AO) and the role of both these ascorbate related enzymes within plant growth and development is imperative. Both AO and APXs are responsible for the conversion of reduced form of ascorbate to an oxidized form (dehydro-ascorbate) which is then transported to cytoplasm for subsequent glutathione mediated reduction to ascorbate finally recycled back to cell apoplast. The biochemical aspects of these two enzymes have been considered in the previous paragraphs in detail while some aspects are considered again. How both of these two enzymes determine substrate specificity or how their regulation takes place can provide insight for discrimination/identification of developmental and stress responses. Here we propose cross talk between AO and APX which can be further integrated with hormonal network.As discussed previously, AO through oxygen initiates ascorbate oxidation in apoplast and this phenomenon is reported to be linked with cell expansion, cell division  which finally decomposes rapidly to dehydro-ascorbate (DHA)  takes place in mitochondria, however, its prime requirement is in the chloroplast Foyer, . It is r− from photosystem II (PSII) to photosystem I (PSI) generates ROS in chloroplasts  to molecular oxygen (O2). This may finally result in severe oxidative stress in the light harvesting organelle and therefore the free radicals are required to be quenched.The light triggered splitting of water molecule in the oxygen-evolving complex (OEC) and subsequent transfer of e Ivanov, . The H2O Kaiser, . In agre Kaiser, . Interes Kaiser,  reported2O2 and ensure sustained light and dark reactions  are primarily the dominant antioxidant enzymes that neutralize chloroplastic HSpinacea oleracea  is known to protect organelles and cells from the adversaries of ROS, which over-accumulate due to stress-induced oxidative damages  to drought stress. In another study with cauliflower plants, Mukhtar et al. (−1) was effective in improving plant growth due to AsA-induced decrease in H2O2 contents and membrane permeability and increase in chlorophyll, proline, GB, potassium and phosphorus contents under water-deficit conditions. Likewise, seed treatment with AsA improved drought stress tolerance in two cauliflower cultivars due to AsA-induced improvement in the activities of catalase (CAT) and (SOD) enzymes in cauliflower plant   which increased AsA level (2–4.25-fold) and ratio of AsA/DHA (3–16-fold) relative to those in wild type Arabidopsis plants. They suggested that increase in plant AsA content through enhanced ascorbate recycling could limit the deleterious effects of stress-induced oxidative stress, particularly temperature stress  is a vital metabolite which is essentially required for the regulation of key physio-biochemical processes in plants. As mentioned earlier, under normal conditions it contributes to a wide range of plant processes, e.g., chloroplast redox reactions, photosynthesis, seed germination, floral induction, fruit expansion, cellular ROS regulation, oxidative defense system, membrane stability, plant water status and senescence etc. Due to its multipurpose involvement within plant cells, its role in plants under stress conditions has become crucial. Several studies have reported the positive influence and stress amelioration by the exogenously supplementation of AsA on plants under various abiotic stress conditions. Varying levels of AsA have been applied during different phases of development to improve stress tolerance in different types of crops including vegetables, oil-seeds, major cereals, pulses, squash, cotton, ornamental/medicinal plants as well as sugarcane. Effectiveness of AsA in the range of 0.1–5.0 mM as well as 10–2000 mg LIn almost all studies reported so far on the role of exogenously applied AsA, synthetic/commercially available AsA have been used. However, extracts of none of the natural plants/fruits, such as papaya, bell peppers, citrus, and straw berries as sources of AsA can been tested to overcome the adverse effects of environmental adversaries on different plants through an eco-friendly source of AsA. Also as a future perspective, the intermediates mentioned in the schematic diagram could be targeted to enhance the AsA contents in plant cells/tissues and modulate subsequent plant stress responses. In this context, it should be investigated that whether the exogenous application of mannose, galactose, gulose and myo-inositol could modulate AsA pools in plant cells and subsequent response to environmental stresses. Similarly, compounds like L-gulono-1,4-lactone and L-galactono-1,4-lactone can also be targeted for improved AsA biosynthesis and investigated their role under stress conditions. Despite the considerable role of AsA in plant stress tolerance, the role of naturally occurring or synthetic analogs of AsA has rarely been focused. Very few studies are available wherein the activities of different enzymes involved in the biosynthesis of AsA and its analogs have been studied, particularly in plants exposed to different stresses. Thus, this needs to be included/elucidated in future research programs.On a biotechnological note, over-expression of the enzymes involved in the biosynthesis of AsA can be targeted to regulate vitamin C contents of plants. There is a need to focus metabolic and biotechnological engineering for improved AsA contents among plants so as to achieve improved tolerance to abiotic stresses. Although the biosynthesis is tightly regulated, the overexpression of certain genes have been successfully achieved in several biofortification studies  and which is again reduced (ascorbate) in cytoplasm by glutathione and then transported back to cell apoplast. In this context, study of apoplast redox status, hormone levels especially auxin and their relationship with NADPH oxidase and comparative regulation of AO and APX activities are essential. These could provide insight into mechanisms of plant environmental stress perception, its amplification and most importantly to discriminate between plant developmental and stress responses.NA wrote the manuscript, FS contributed in section introduction of this manuscript, while MA reviewed and updated the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Acute pancreatitis (AP) is a common abdominal acute inflammatory disorder and the leading cause of hospital admission for gastrointestinal disorders in many countries. Clinical manifestations of AP vary from self-limiting local inflammation to devastating systemic pathological conditions causing significant morbidity and mortality. To date, despite extensive efforts in translating promising experimental therapeutic targets in clinical trials, disease-specific effective remedy remains obscure, and supportive care has still been the primary treatment for this disease. Emerging evidence, in light of the current state of pathophysiology of AP, has highlighted that strategic initiation of nutrition with appropriate nutrient supplementation are key to limit local inflammation and to prevent or manage AP-associated complications. The current review focuses on recent advances on nutritional interventions including enteral versus parenteral nutrition strategies, and nutritional supplements such as probiotics, glutamine, omega-3 fatty acids, and vitamins in clinical AP, hoping to advance current knowledge and practice related to nutrition and nutritional supplements in clinical management of AP. Acute pancreatitis is the leading cause of acute hospital admission for gastrointestinal disorders in many countries, and its incidence continues to raise worldwide –3. The aPathophysiology of AP encompasses complex cascaded events of acinar cell inflammation, involvement of immune system, and systemic pathological outcomes  Figure . PrematuBetter understanding of the pathophysiology of AP has drawn research efforts to reestablish the immune and organ/tissue homeostasis in clinical AP and toward the development of new intervention strategies . With stnothing by mouth) strategy has been implemented conventionally to treat AP , glutamine, ω-3 FAs, and vitamins, hoping to provide the basis for future development of nutritional strategies in clinical AP.nil per os or nothing per mouth treatment until resolution of pain or normalization of pancreatic enzymes to allow the pancreas to rest . Patients receiving synbiotic therapy had reduced total incidence of SIRS and lower rates of organ failure, supporting that early EN with synbiotics may prevent organ dysfunctions in the late phase of severe AP  consumption may be associated with decreased risk of non-gallstone-related AP . A randoP < 0.05)  was not justified to continue in clinical severe AP  . Recentl < 0.05) . Vitaminevere AP . Subsequal trial . With real trial . In contal trial . CollectIn most patients, an oral soft or solid diet can be beneficial if tolerated. When oral feeding is not tolerated for a few days, enteral feeding through a nasogastric or nasojejunal feeding tube should be attempted within the first 72 h of administration. PN should be minimized for its risks of infection and other complications. Only if enteral route is not available or tolerated, PN may be considered. Overall, nutritional support plays a critical role in clinical management of severe AP, although the optimal timing remains unclear. Predicting the nutritional tolerance of patients with AP remains challenging as the current evaluation system needs to be improved. Various nutritional supplement(s) together with PN or EN with currently mixed clinical outcomes is a subject of interest for future evaluation and may lead to promising outcomes. In addition, given its heterogeneous etiological factors and varying clinical manifestations, precision medicine, although not much applied in the condition, remains as a temping approach to optimize clinical outcomes on classified individuals based on susceptibility to the condition and its systemic complications.JS designed the subject content of the review article. L-LP, JL, MS, and JS conducted initial search of literature, drafted the manuscript, and prepared the figures and tables. MB gave the constructive comments and critically reviewed the manuscript. JS had primary responsibility for final content. All authors read and approved the final 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.


Despite investment in family planning programs and education, unmet need for family planning remains high among young women (aged 15–24) in low and middle-income countries, increasing the risk for unwanted pregnancies and adverse social and reproductive health outcomes. There is a dearth of cross-national research that identifies the differential impact of community level factors among youth in low and middle-income countries (LMICs), which is imperative for the design of structural level interventions aimed at increasing family planning use.Grounded in the socio-ecological framework, this paper utilizes Demographic and Health Survey (DHS) from 52 LMICs to examine the influence of community level reproductive, gender, fertility, literacy and economic indicators on modern contraceptive use among female youth. Analyses are conducted using multi-level logistic regressions with random community-level effects.Our findings highlight the positive influence of community level education attainment and negative influence of gender and fertility related norms on young women’s contraceptive use. Additionally, increased exposure to mass media did not positively influence young women’s uptake of modern contraceptive methods.Taken together, findings indicate that young women’s contraceptive decision-making is greatly shaped by their social contexts. The commonalities and regional variations in community level influences provide support for both structural level interventions and tailored regional approaches to family planning interventions. Family planning is one of the most important health interventions of the twentieth century . It enabContraceptive use among young women aged 15–24), whether married or unmarried, is lower than among relatively older women in the developing world. Consequently, over 39.5% of young unmarried women and 25.9% of young married women in low and middle-income countries have an unmet for family , Eastern Mediterranean region [AOR: 1.08], European region [AOR: 1.17], the Americas [AOR: 1.05] and South East Asia [AOR: 1.16] but associated with lower odds for modern contraceptive use among young women in the Western Pacific region [AOR: 0.88].Despite ongoing efforts to expand access to family planning services, unmet for family planning among young women remains high . UnderstWith a few notable exceptions, our findings on the global association between the community factors and contraceptive use were largely consistent with the current literature , 19, 20.Regional patterns in the associations between the community level factors related to mean age at first marriage, mean age at first birth, mean age at sexual debut and exposure to mass media, education attainment of women, attitudes towards intimate partner violence and contraceptive use also emerged. These findings emphasize the contextual nature of the interrelationship between community demographics, socio-cultural norms, and gender dynamics with contraceptive use. These findings also highlight the need to considering both space and time in investigations of community level influences on contraceptive behavior because communities and young women occupy spaces that are continually evolving with political, cultural and social discourses that are unique to each space and time , 29. YouOf the several community level factors examined in this paper, the mean ideal family size emerged as the most salient community level predictor of modern contraceptive use among young women. In all regions, young women living in communities with a greater ideal number of children were less likely to report using a modern contraceptive method. This finding could be attributed to the pro-natal attitudes that are prevalent across low and middle-income countries , 32. TheThe study has a number of limitations. First, the cross-sectional nature of the DHS data precludes analyses of causal relationships between the community level factors and contraceptive use. Second, the data are heavily weighted towards countries in Africa; as such, they may not provide an accurate representation of other regions that have fewer countries represented within the DHS. Third, as discussed, the PSUs may not be representative of natural community formations and represent artificial sampling blocks. The analysis did not incorporate infrastructural factors , peer-related factors, and ethnic norms/practices that could influence young women’s contraceptive behavior as this is not collected in the DHS. Fourth, our analyses are based on self-report data, which is vulnerable to recall bias and social desirability bias. Some authors have also raised concerns about the quality of data in earlier DHS surveys, particularly with regard to indicators of sexual activity among youths SSA ; this coThese findings have numerous implications for programs and research aimed at increasing uptake of modern contraceptive methods among women in sub-Saharan Africa. First, they highlight the potential mechanism through which community level factors may shape young women’s contraceptive use. In particular, these highlight the need for community-wide multifaceted programs to address the persistent influence of socio-cultural and gendered norms on uptake of modern contraceptives. Such programs should be implemented in tandem with greater efforts to expand young women’s access to education and employment opportunities, youth friendly sexual reproductive health services, comprehensive health information and life skills training, to promote positive decision-making.Second, they underscore the need for tailoring and audience segmentation in family planning programs. As indicated, they findings suggest that community level factors may shape contraceptive use in younger and older women differently. Audience segmentation is an important health tool that allows programs to identify relatively homogenous sub-populations with similar characteristics and needs that can be targeted through tailored messages and programs that meet these needs , 38.Lastly, these findings underscore the need for longitudinal studies to enhance our understanding how the dynamic interactions between space and time may shape contraceptive use during the transition to adulthood. Currently, our understanding of contraceptive use among young women is informed by data from cross-sectional studies: this limits our ability to unpack the complexities in women’s experiences, particularly in the context of competing and, at times, conflicting interrelationships between the individual and community level factors that shape women’s FP decision-making processes and experiences. As such, our findings highlight the need for further studies, particularly qualitative and longitudinal studies, to expand our understanding of the complex inter-play between the various individual and community factors that shape women’s contraceptive behavior and how these vary over the life course.


Glioblastomas are devastating central nervous system tumors with abysmal prognoses. These tumors are often difficult to resect surgically, are highly invasive and proliferative, and are resistant to virtually all therapeutic attempts, making them universally lethal diseases. One key enabling feature of their tumor biology is the engagement of the unfolded protein response (UPR), a stress response originating in the endoplasmic reticulum (ER) designed to handle the pathologies of aggregating malfolded proteins in that organelle. Glioblastomas and other tumors have co-opted this stress response to allow their continued uncontrolled growth by enhanced protein production (maintained by chaperone-assisted protein folding) and lipid biosynthesis driven downstream of the UPR. These features can account for the extensive extracellular remodeling/invasiveness/angiogenesis and proliferative capacity, and ultimately result in tumor phenotypes of chemo- and radio-resistance. The UPR in general, and its chaperoning capacity in particular, are thus putative high-value targets for treatment intervention. Such therapeutic strategies, and potential problems with them, will be discussed and analyzed. Glioblastoma multiforme (WHO grade IV glioma [GBM]) is a devastating tumor of the brain/central nervous system, and is also the most common malignant primary brain tumor in the USA. Patients with GBMs seldom live longer than 15 months . The conSolid tumors are existentially stressed tissues. Their disregard for normalizing cues concerning unrestrained cell proliferation leads to inadequate blood supplies, insufficient nutrients and a hypoxic environment. The host's immune system, while largely inadequate, nonetheless attacks the tumor and requires resistance, and exogenously we attempt to inflict damage with chemical agents and radiation. These stresses provoke tumors to defend themselves by upregulation of particular pathways designed for cytoprotection, particularly those involved in protein folding and protein stabilization. These pathways include upregulation of chaperone proteins/heat shock proteins in the cytoplasm (and other organelles such as the mitochondria and endoplasmic reticulum), including those involved in the unfolded protein response (UPR) ,4. GBMs Ostensibly, the UPR manifests when misfolded proteins accumulate in the endoplasmic reticulum (ER) , but theThe UPR is a beautifully complex set of molecular interactions that globally alter cellular transcriptional and translational profiles, involving sensors, transducers and multiple layers of effectors. One of the ER sensors is the HSP70 family member GRP78 (BiP/HSPA5); in its chaperone role it can detect unfolded/malfolded proteins in the ER lumen, and can assist in their folding in an ATP-dependent manner . Under cWith GRP78 bound to these three ER membrane proteins, they remain in a monomeric/tethered state; once some triggering level of unfolded proteins appears in the ER lumen, GRP78 releases the three to engage in chaperone duties for the unfolded proteins. PERK and IRE1 now can dimerize or oligomerize, and ATF6 simply leaves the organelle, to act as transducers of the UPR . The diATF4 are translated more efficiently in this situation , reducing the GDP-GTP exchange of eIF2-GTP. This prevents binding of eIF2 to initiator methionine tRNAs and loading into the P site on the 40S ribosome, effectively halting most translation . Howeverart site ). In addart site , such asart site . Some ofart site ), XBP-1 art site . Howeverart site ; thus, tXBP-1 mRNA , and allows splicing (by unknown ligases) to generate the longer  mRNA with a frame shift that now codes for a longer, more stable and active protein transcription factor. IRE1 is also responsible for degradation of other mRNAs, particularly those encoding membrane and secreted proteins )Downstream of both IRE1 and ATF6 is XBP-1. Direct inhibition of XBP-1 transcriptional activities has not been described. Alternatively, the triene-ansamycins were identified as compounds preventing the splicing of XBP-1 mRNA . None ofConceptually, ATF6 might be inhibited directly, but no compounds currently seem to do this – however, such screening may be underway . ATF6 coIn general, pharmacologic inhibition of transcription factors has historically been considered difficult, usually due to the lack of enzymatic activities associated with such proteins . Thus, dAs mentioned above , ERAD isOur recent metabolomic work showed that recurrent GBMs produce more lipids of almost every category compared with primary GBMs, and those recurrent tumors have high lipogenesis enzyme expression . As therThe stressful tumor environment drives stress responses in tumors that allow for their adaptation to this environment. The UPR is one such response that appears to benefit the ‘stressed’ tumor by enabling extended protein folding capabilities with high chaperone content, enhanced lipid biosynthesis for extracellular remodeling and generating metabolic profiles consistent with drug resistance  to deterThe unfolded protein response (UPR) is an ancient endoplasmic reticulum (ER)-based cytoprotective mechanism evolved to protect cells from pathologic protein aggregation. It engages three arms to halt incoming protein load into the ER, to aid in protein folding of the molecules in the ER, to increase shipments out of the ER, to degrade excess unfolded proteins, and to initiate cell death if the stress is intractable.Tumor cells, especially those from high grade gliomas/glioblastoma multiforme (GBMs), engage the UPR in various ways, perhaps initially as a stress response, but often seemingly to benefit from the increased outputs of proteins and lipids, without suffering the apoptotic consequences.The high chaperone protein content of GBMs (and other brain tumors) implies that the folding pathways and the antiapoptotic pathways involving chaperones are critical features of such tumor biology.These features, as well as those intertwined with the UPR, appear as provocative targets for drug intervention. However, to date few, if any, of those pharmacologics have had any significant impact on treatment regimens for patients with GBMs. We need further refinements in our understanding of tumor stress biology and its consequences for tumor survival and progression to appropriately design effective treatment strategies.


Anaemia is the most frequent, though often neglected, comorbidity of inflammatory bowel disease (IBD). Here we want to briefly present (1) the burden of anaemia in IBD, (2) its pathophysiology, which mostly arises from bleeding-associated iron deficiency, followed by (3) diagnostic evaluation of anaemia, (4) a balanced overview of the different modes of iron replacement therapy, (5) evidence for their therapeutic efficacy and subsequently, (6) an updated recommendation for the practical management of anaemia in IBD. Following the introduction of various intravenous iron preparations over the last decade, questions persist about when to use these preparations as opposed to traditional and other novel oral iron therapeutic agents. At present, oral iron therapy is generally preferred for patients with quiescent IBD and mild iron-deficiency anaemia. However, in patients with flaring IBD that hampers intestinal iron absorption and in those with inadequate responses to or side effects with oral preparations, intravenous iron supplementation is the therapy of choice, although information on the efficacy of intravenous iron in patients with active IBD and anaemia is scare. Importantly, anaemia in IBD is often multifactorial and a careful diagnostic workup is mandatory for optimized treatment. Nevertheless, limited information is available on optimal therapeutic start and end points for treatment of anaemia. Of note, neither oral nor intravenous therapies seem to exacerbate the clinical course of IBD. However, additional prospective studies are still warranted to determine the optimal therapy in complex conditions such as IBD. Anaemia and iron deficiency are global health issues and a recent analysis estimated that approximately one-third  of the world’s population is anaemic . Furthern = 775) or ulcerative colitis (UC) (n = 512) revealed that flaring disease is the parameter most consistently related to the presence of anaemia, with no differences between CD and UC, although anaemia is more frequent among women [p < 0.001) [In inflammatory bowel disease (IBD) ,8, whichng women , especiang women . Further< 0.001) . It was < 0.001) . Moreove< 0.001)  and 68% < 0.001) , exceedi< 0.001) ,19.The pathogenesis of anaemia in IBD is multifactorial and results mainly from intestinal blood loss in inflamed mucosa and impaired dietary iron absorption . The chrApart from IBD, anaemia is observed in a number of chronic inflammatory disorders. These include autoimmune disorders , cancer and infections. This so-called anaemia of chronic disease (ACD) or anaemia of inflammation is more prevalent in patients with advanced disease and in those responding poorly to therapy .Treatment of iron-deficiency anaemia in IBD is of importance because of the possible consequences on multiple organs and biological processes. These include cellular dysfunctions comprising impaired mitochondrial respiratory capacity and metabolic impairments that translate into specific organ dysfunctions, for example, in the central nervous system , immune system , cardiorespiratory system , vascular system , genital tract  and gastrointestinal tract  ,33. CollThe perception that anaemia in IBD needs specific treatment apart from regular control of IBD is still underdeveloped. Thus anaemia is neither diagnostically worked up nor do the majority of anaemic patients receive any specific treatment , althougIron constitutes a key part of haemoglobin in erythrocytes and of myoglobin in muscles, which in combination contain approximately two-thirds of the total body iron. In addition, iron is crucial to a wide range of biological processes ,42. The 2+) to the protoporphyrin ring, which is abundant in animal food products such as meat, poultry and seafood [3+ or ferric iron) and is present in foods of vegetable origin . Heme iron is assumed to constitute 10–15% of total iron intake in meat-eating populations but because of its higher bioavailability (estimated absorption rate of 15–35%) than nonheme iron (5–15%), it accounts for more than 30% of the total absorbed iron [Dietary iron is available in two forms: heme and non-heme-bound iron. Within heme, iron is complexed as ferrous iron  with animal data showing some absorption from the large bowel as well [3+. Circulating iron forms complexes with transferrin and is delivered as transferrin-bound iron to cells and tissues. Most of the iron needed for metabolic purposes and erythropoiesis (approximately 20–30 mg/day) originates from macrophages that engulf senescent erythrocytes and reuse iron that is returned to the circulation via ferroportin. During systemic inflammation and increased levels of inflammation-induced hepatic hepcidin secretion, the iron transporter ferroportin found in cells of the reticuloendothelial system and in enterocytes is degraded and cellular iron export is reduced. This results in iron retention in cells of the reticuloendothelial system and impaired dietary iron absorption, subsequently resulting in low serum iron levels with all the clinical consequences mentioned in the text. To overcome iron deficiency in patients with IBD, iron supplementation in the form of oral or intravenous iron can be applied. Novel approaches include inhibition of hepcidin itself or its expression. The most important preventive intervention for long-term well-being of the patients, however, is to efficiently treat the underlying condition, in this case the intestinal inflammatory process. The asterisk indicates points of therapeutic intervention. Between 1 and 2 mg of iron is taken up daily from the diet, which is balanced by its secretion mainly through intestinal and skin epithelial desquamation. Intestinal bleeding in patients with IBD increases iron loss. With the majority of iron being taken up in the duodenum through heme and nonheme iron transporters  or anaemia of inflammation by retaining iron in the reticuloendothelial system and blocking iron absorption, causing an iron-limited erythropoiesis ,55. ThusHowever, patients with flaring IBD experience chronic blood loss due to intestinal mucosal bleeding, which often causes true iron deficiency in conjunction with inflammatory anaemia  ,56. Of n12, folic acid and vitamin D) due to either intestinal inflammation or extensive bowel resection can also contribute to the development of anaemia [Finally, vitamin deficiencies , adult males and females with a blood haemoglobin concentration below 13 and 12 g/dL, respectively, are considered anaemic (<11 g/dL during pregnancy) . ThreshoAs a consequence of chronic inflammation, patients with active IBD may show reduced levels of transferrin, which is contrary to the definition of patients with iron deficiency . ImportaSerum ferritin concentration, which generally correlates with body iron stores, is the most widely used surrogate marker of stored iron and works nicely in patients without any concomitant inflammatory condition. Circulating ferritin levels, however, are influenced by inflammation; in fact, several proinflammatory cytokines stimulate ferritin expression, leading to measurements in the normal or even elevated range during chronic inflammation, even in the presence of true iron deficiency ,72,73. TSerum-soluble transferrin receptor (sTfR), a proteolytic derivative of the membrane-bound transferrin receptor, is another marker of iron status. With true iron deficiency, increased synthesis of transferrin receptors is observed along with a corresponding increase in sTfR levels. Nevertheless, the sTfR concentration might also rise during disorders associated with increased erythropoiesis, including chronic lymphatic leukaemia, whereas it can be reduced by the actions of cytokines during inflammation. Therefore, no consensus currently exists as to a standardized cut-off value for sTfR . To distBased on erythrocyte analyses of full blood samples from anaemic patients, information about mean cellular haemoglobin concentration  and mean cell volume  can be obtained. The values of these indices are decreased by iron deficiency. In the case of microcytosis in patients with an appropriate ethnic background, haemoglobin electrophoresis may be considered to rule out hemoglobinopathies such as sickle-cell disease and thalassemia . MicrocyBone marrow aspiration for diagnosing iron deficiency appears to be the gold standard. This method is thought to be unaffected by inflammation but it is invasive in nature, uncomfortable for the patient, expensive and might be affected by concomitant treatment with recombinant erythropoietin. Thus, bone marrow aspiration should be reserved for specific situations where other techniques are either unavailable or conflicting .Hepcidin has a key regulatory role in iron homeostasis as an inhibitor of cellular iron export . MeasureIt is of great importance to establish the presence of true iron-deficiency anaemia in patients to avoid any unnecessary treatment. The assessment should be guided by predictive serum parameters such as hepcidin, soluble transferrin receptor and others including red cell indices but no gold standard is available at present ,84. The The primary treatment of ACD is to cure the underlying pathology or other easily treatable conditions contributing to anaemia, such as vitamin deficiency, which often leads to improvement in haemoglobin levels unless other pathophysiologic factors or deficiencies are present ,29,33,85Imbalances of iron homeostasis are the major reason for anaemia in patients with IBD. The currently available options for iron supplementation to balance iron intake and iron loss consist of oral and intravenous administration and their pros and cons are listed in 2+ (ferrous) or trivalent Fe3+ (ferric) salts coupled with sugar complexes or protein succinylate [2+)—A process catalysed by membrane-bound ferric reductase. Divalent metal transporter-1 (DMT1) facilitates iron uptake in the acidic environment [3+ to Fe2+ prior to epithelial uptake [The bioavailability of “traditional” oral iron preparations is relatively low but nevertheless is the first-line therapy in iron-deficiency anaemia. Oral iron has a well-established safety profile, is easy to administer and comes with a generally low cost—with the latter being important in a pharmaco-economical setting . Oral ircinylate ,96. The ironment . Ascorbiironment  by provil uptake . Furtherl uptake  D) below 30 ng/mL  were associated with increased hepcidin concentrations and reduced haemoglobin concentrations compared with individuals with plasma 25(OH)D concentrations above 30 ng/mL . Because3+ reduction by the enzyme ferric reductase to enhance its activity. Vitamin C also suppresses the inhibitory features of phytate and calcium on iron uptake. Vitamin D obtained from the diet or generated in the skin through ultraviolet-induced photolysis of vitamin D precursors augments iron absorption by lowering mRNA expression of hepcidin mediated by the presence of vitamin D response elements (VDREs) identified in the promoter region of the hepcidin gene. Additionally, vitamin D inhibits the release of IL-1 and IL-6 and increases erythroid progenitor proliferation.Dietary vitamin C enhances iron absorption by providing reducing equivalents for FeRecently, new orally available products have been introduced into clinical practice. One of these, ferric maltol, has been successfully studied in a phase III trial in IBD patients with iron-deficiency anaemia who had previously been either intolerant or unresponsive to oral ferrous products .Although the optimal dose of oral iron supplements in patients with IBD and iron deficiency has not been established, a dose of 50–200 mg/day of elemental iron is often recommended . Only 10Given the low absorption of oral iron, a high proportion remains in the gut and is associated with the development of gastrointestinal side effects, including nausea, dyspepsia, diarrhoea, abdominal discomfort, vomiting and constipation in up to 20% of patients—often resulting in nonadherence to therapy ,114. GenMost of the reservations regarding oral iron therapy in IBD come from studies in animal models, which have shown contradictory evidence regarding the impact of oral iron on ongoing intestinal inflammation ,116. In Parenteral iron administration more rapidly increases haemoglobin levels than oral delivery ,123 and Six intravenous iron preparations are available at present. These include iron dextran, iron gluconate and iron sucrose, as well as the more recently licensed high-molecular-weight compounds ferumoxytol, iron isomaltoside 1000 and ferric carboxymaltose ,130,131.The iron dextran compounds exist in two forms that are stable: A low- (73 kDa) and a high-molecular-weight (165 kDa) complex. Because the latter has been linked to an increased risk of both anaphylactic and anaphylactoid reactions ,134,135,The stability of both iron gluconate (37 kDa) and iron sucrose (43 kDa) is lower than that of iron dextran and these two iron compounds can be administrated at a maximal single dose of 200 mg of iron gluconate (300 mg in some countries) over a minimum infusion time of 30 min  or 62.5 The introduction of more stable iron complex formulations for intravenous iron administration has permitted the infusion of higher single doses with minimal side effects and no need for test doses because of the marginal release of labile iron during administration. The highly stable 150 kDa complexes ferric carboxymaltose ,144,145 Although different side-effect profiles are associated with various preparations of large-molecule iron complexes , the mosBased on our expanding knowledge of the pathways underlying inflammatory anaemia and specifically the role of hepcidin, new therapeutic strategies are emerging that attempt to block hepcidin activity either by directly interfering with hepcidin synthesis by affecting different inflammation- or iron-driven signalling pathways that regulate hepcidin expression  or by neutralizing hepcidin in the circulation ,159,160 Members of our group previously performed a systematic search that yielded a total of 632 studies concerning iron therapy in IBD published from January 2004 to March 2015 , of which 13 prospective trials met the inclusion criteria as randomized, controlled trials and included 2906 patients in total . This syComparative studies of intravenous versus oral iron supplementation in the systematic review did not demonstrate any significant difference in haemoglobin normalization favouring the use of intravenous iron therapy unless considered for patients with intolerance or an inadequate response to oral supplementation . In patiIt is known that apart from the WHO definitions of anaemia , a low Tp = 0.016) at the same time [Only nine randomized, controlled trials investigating oral iron supplementation in IBD patients were published between 2004 and 2017 ,145,165.ame time  and whename time , an obseA very recent systematic review and Bayesian network meta-analysis performed on the five eligible randomized, controlled trials with a total population of 1143 patients has shown ferric carboxymaltose to be the most effective preparation for the treatment of iron-deficiency anaemia in IBD, followed by iron sucrose, iron isomaltoside and oral iron in fourth place . This anIt is generally accepted that individuals with iron deficiency and coexisting anaemia need treatment. It is, however, a subject of debate whether treatment of iron deficiency should be initiated before the development of anaemia—a condition that recently was reported to occur in 37% of IBD patients in a Spanish outpatient cohort —because The cause of anaemia and specifically of concomitant iron deficiency should be identified in every patient with IBD. Thus, the recurrence of iron deficiency following successful treatment is often due to persistence or relapse of the initial inciting cause , which should be managed appropriately. Further, in patients who have failed to respond to either oral or parenteral iron therapy, the cause for this failure should be carefully determined.Previously it was accepted that clinical symptoms of anaemia occurred only when the haemoglobin level dropped abruptly  and, conClinical guidelines often emphasize that because of the ease of treatment, patients with uncomplicated iron-deficiency anaemia should be treated with oral rather than intravenous iron formulations . In thisIndications for intravenous iron administration include severe anaemia (haemoglobin < 10 g/dL), intolerance of or inappropriate response to oral iron administration, severe intestinal disease activity and concomitant therapy with an erythropoiesis agent or patient preference. Oral iron supplements can be used if these indications for intravenous therapy are not met.If intravenous iron supplementation is considered, the use of low-dose regimens is not recommended from the point of view of clinical efficacy because a number of infusions might be needed over several days or weeks. Instead, high-dose regimens that result in fewer infusions and increase both convenience and cost-effectiveness of intravenous iron repletion should be considered.The optimal dosing strategy for intravenous iron compounds depends on the type of preparation, the body weight of the patient and the haemoglobin concentration. The amount of iron needed to correct the haemoglobin can be calculated using the Ganzoni equation , often rLast but not least, it should be kept in mind that iron deficiency in IBD often relapses after iron replenishment . ConsequThis leads to the questions of (1) therapeutic start and end points  and (2) whether or not iron-deficiency anaemia should be treated differently depending on the underlying disease? To start with the latter point, in general, subjects with pure iron-deficiency anaemia on the basis of an inadequate dietary iron intake and/or increased blood losses and iron-deficient IBD patients with no or minimal signs of inflammation should initially be recommended to oral iron therapy. However, such a recommendation might have some caveats. Oral iron replacement therapy may be of limited efficiency in the setting of concomitant inflammation, which is usually associated with increased hepcidin concentrations resulting in an impaired response to iron therapy ,187. YetWhile normalization of haemoglobin appears to be a reasonable readout in subjects with iron-deficiency anaemia in the absence of inflammation , retrospImportantly, life-threatening reactions possibly caused by release of free iron are rare after administration of intravenous iron supplementation therapies ,193. ThuPatients with inflammatory diseases will respond poorly to oral iron therapy unless the iron deficiency is severe. Newer iron formulations, such as ferric maltose, have been shown to correct mild anaemia in patients with quiescent IBD  but whetHere we have summarized the impact and pathophysiology of iron deficiency in the setting of IBD. Diagnostic criteria are provided as well as methods to differentiate between functional and true iron deficiency. We also discussed the currently available drugs and commented on issues that should be considered by physicians treating patients with IBD. Thus, treating physicians need to pay more attention to the management of anaemia and iron deficiency for improvement of the general well-being of their patients with IBD—a matter that actually does not gain the attention it deserves. Although we lack knowledge on the effects of iron repletion strategies on the course of IBD, the control of inflammation is pivotal in the management of anaemia in this intestinal disorder.Given the novel intravenous high-dose iron replacement regimens introduced within the last decade, oral iron therapy should be preferred for IBD patients with mild and uncomplicated iron-deficiency anaemia (haemoglobin ≥ 10 g/dL) in quiescent disease stages unless previous complications have been observed, including an inadequate response (haemoglobin increase < 2 g/dL within 4 weeks) . IntraveFinally, it should be emphasized that iron deficiency may relapse often after iron replenishment , specifi


The ends of linear genomes, whether viral or cellular, can elicit potent DNA damage and innate immune signals. DNA viruses entering the nucleus share many features with telomeres in their ability to either suppress or co-opt these pathways. Here, we review some of the common mechanisms that viruses and telomeres use to manage the DNA damage and innate immune response pathways. We highlight recent studies on the role of the telomere repeat-containing RNA (TERRA) in response to viral infection. We discuss how TERRA can be activated through a p53-response element embedded in a retrotransposon-like repeat found in human subtelomeres. We consider how TERRA can function as a danger signal when secreted in extracellular vesicles to induce inflammatory cytokines in neighboring cells. These findings suggest that TERRA may be part of the innate immune response to viral infection, and support the hypothesis that telomeres and viruses utilize common mechanisms to maintain genome integrity and regulate innate immunity. Viruses and telomeres share numerous structural and molecular features that require the competitive utilization of common factors required to propagate and maintain their respective genomes . For thecis and other genetic loci in trans by interacting with a large network of proteins, including epigenetic and telomeric factors [Telomeres are repetitive elements at the ends of linear chromosomes that are essential for maintaining genomic stability ,6,7. Tel factors . TERRA t factors . Subtelo factors . These r factors . Nucleosome assembly and chromatin organization are critical for genome maintenance and protection against the various DNA damage sensing and repair pathways. Telomeres have a complex chromatin structure, are typically enriched with heterochromatic features, and are localized to the periphery of the nucleus . The hisTelomeres have been shown to have a complex 3D organizational structure that can affect chromatin and gene expression at subtelomeric sites ,31,32. TISG15), an interferon-regulated gene that is among the most significantly activated upon viral infection. Thus, telomere repeat length and telomeric chromatin have functional links to innate immune signaling pathways, and cells with different telomere lengths may have different responses to viral infection. Telomere repeat length may also contribute to telomeric heterochromatin and affect local transcription through telomere DNA-looping into the subtelomeres . InteresDNA virus infection in the nucleus can activate a DDR signaling pathway similar to that of a chromosome double strand break or an uncapped telomere . Like teAn alternative mechanism to evade double-strand break recognition is observed in herpesviruses that enter the nuclei as linear genomes, but then circularize at early time points of the viral life cycle ,42,43. CViral DNA is recognized by the promyelocytic leukemia-nuclear bodies (PML-NBs) , an interferon-inducible nuclear structure consisting of core proteins PML, SP100, Daxx, and ATRX ,56,57,58PML-NBs have been implicated in the ALT ,68,69. HIn addition to its anti-viral role and function in ALT, PML has been implicated in telomere maintenance in diploid fibroblasts . The siRn, suggesting that it has the capacity to recognize telomeres [Other cellular factors have been implicated in the intrinsic response to naked viral DNA in the nucleus. Among the first nuclear factors that recognize and respond to invading naked DNA is the Interferon-Inducible 16 (IFI16) protein, a member of the PYHIN family of DNA binding proteins  secreted from cells to mediate intercellular communication and deliver functional factors . Cell-frThe release of these telomeric factors has several impacts on the microenvironment. cfTERRA-containing exosomes were a strong inducer of inflammatory cytokines, including interleukin (IL)6 and tumor necrosis factor (TNF)α . The incViruses also hijack exosome signaling to evade host immunosurveillance. Cells latently infected with EBV type III latency had high levels of cfTERRA . Type IIExosomes share several features with those of virus-like particles. Like viruses, exosomes are infectious particles containing protein and nucleic acids, but an important difference is that exosomes lack the capacity to replicate. However, their structural and functional similarity to viruses suggests that they share similar evolutionary origins. Viruses such as HIV-1 and the Hepatitis E virus utilize similar pathways as exosomes to facilitate viral infection and virion release ,117. SevThe analogies between viruses and telomeres may be a useful framework to understand similar genetic and genomic processes, and act as a heuristic guide in developing new hypotheses to test experimentally or through bioinformatic forensics. Here, we have considered how viruses can resemble uncapped telomeric DNA, and share the strategies to evade DNA damage recognition and degradation. We have discussed the role of p53 response elements in subtelomeres and how these may affect telomeric adaptation to DNA damage stress, which is perhaps similar to how viruses may reactivate during latent infection. We have also considered how telomeres can express and package TERRA to communicate with neighboring cells, and how this resembles viral packaging and transmission. While some of the common mechanisms and themes are not limited to viruses or telomeres, and there is no surprise that viruses have captured many cellular processes, there remain compelling arguments that the common features of viruses and telomeres have emerged from either common ancestry or shared biological constraints . In eith


Group B streptococcus (GBS) is a leading cause of neonatal mortality and morbidity in the United States and Europe. It is part of the vaginal microbiota in up to 30% of pregnant women and can be passed on to the newborn through perinatal transmission. GBS has the ability to survive in multiple different host niches. The pathophysiology of this bacterium reveals an outstanding ability to withstand varying pH fluctuations of the surrounding environments inside the human host. GBS host pathogen interations include colonization of the acidic vaginal mucosa, invasion of the neutral human blood or amniotic fluid, breaching of the blood brain barrier as well as survival within the acidic phagolysosomal compartment of macrophages. However, investigations on GBS responses to acid stress are limited. Technologies, such as whole genome sequencing, genome-wide transcription and proteome mapping facilitate large scale identification of genes and proteins. Mechanisms enabling GBS to cope with acid stress have mainly been studied through these techniques and are summarized in the current review Streptococcus agalactiae or group B streptococcus (GBS) is an opportunistic pathogen which colonizes up to 30% of the genitourinary and gastrointestinal tracts of healthy women. At the same time GBS is a leading cause of life-threatening neonatal infections, such as meningitis, sepsis and pneumonia , transcriptional regulators and sigma factors which respond to acid stress by modifying gene expression. Moreover, metal ion homeostasis, osmoregulation and oxidative stress response have been increasingly reported to contribute to acid adaptation mechanisms.Low pH environments are often encountered by GBS inside the human host and mechanisms that enable GBS to cope with acid stress must therefore be essential for colonization as well as infection. However, our understanding of the acid stress response in GBS is incomplete. While the relevant mechanisms have often been studied at a functional level in other streptococci or gram positive pathogens studies in GBS largely rely on genomic and transcriptomic investigations. With this review, we try to summarize our current knowledge about mechanism permitting GBS survival at low pH and draw comparisons to other gram positive bacteria, especially streptococci. Figure A low pH of 4 ± 0.5 is crucial to maintain a healthy vaginal environment. These pH-values mediate a microbicidal impact, which has been demonstrated to be effective against various microbial pathogens of sexually transmitted diseases (STD), including HIV , and urinary tract infections (UTIs)  intraluminal pH. As maturation proceeds to later stages, the lumen becomes more acidic (pH 5.5–6.0) and enriched in proteases and lysosomal-associated membrane proteins (lAMPs). Terminal phagolysosomal compartments are strictly acidic, possessing intraluminal pH-values as low as 4.5, and contain reactive oxygen species (ROS), reactive nitrogen species (RNS), and antimicrobial peptides. Thus, during the maturation process, phagosomes become fully armed with destructive antimicrobial features , it comprises an arginine deiminase, ornithine carbamoyltransferase , and carbamate kinase. These three enzymes are encoded by the S. pyogenes  has been characterized in oral streptococci   are composed of a membrane-associated histidine kinase sensor and a cytoplasmic response regulator. They contribute to adaptation, virulence and survival through detecting environmental fluctuations and providing a proper response , GroEL and GroES, encoded by the GroE operon (groES-groEL), and Clp proteases , such as superoxide anions, hydrogen peroxide and hydroxyl radicals. ROS are toxic leading to irreversible DNA and protein damage  is another system that is involved in protection against oxidative stress. This system has been shown to be essential for the respiratory metabolism and establishing a full respiration chain in GBS when the surrounding environment supplies heme and quinone. Aeration alone had a slight effect on aerobic growth in comparison to static conditions in GBS as indicated by Yamamoto et al. . Interesi et al.  reportedS. mutans, peroxidase activity is mainly achieved through the AhpC-AhpF complex where AhpF (or Nox-1) acts as a dehydrogenase that reduces NADH and delivers electrons to AhpC. In addition, AhpC converts peroxidase into water or alchohol  and thiol peroxidase (Tpx) (Lemos et al., 2+ and Fe2+, the Mn-dependent SOD, cytochrome bd quinol oxidase and thiol peroxidase. These genes can be targeted in further mutagenesis studies to assess and confirm their physiological significance for GBS in coping with acid stress. As discussed above, a successful scenario was recently shown when proofing the direct contribution of the mntH NRAMP transporter in GBS acid stress that was first mentioned in an earlier genome-wide transcription study.In GBS a wealth of data has accumulated from genome sequencing projects, genome-wide transcription analysis and proteome mapping resulting in the large-scale identification of genes and proteins that are induced in response to low-pH environments. However, care must be taken when interpreting these results, to differentiate between genes and proteins with a direct contribution in the pH stress response and more unspecific effects. To validate these results, more conventional techniques need to be employed, such as the construction of mutants and addressing the consequences of these mutations on an individual basis. Unfortunately, functional studies to investigate GBS acid responses are still rare. Nonetheless, this review shows that genomic and proteomics-based approaches have identified many acid-inducible genes, such as the ADI system genes, F1F0-ATPase genes, genes of the glycine betaine system, genes encoding Clp chaperones, metal-ion transporters especially for MnBS and SS both designed and wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Earlier research primarily attributed the effects of mesenchymal stem cell (MSC) therapies to their capacity for local engrafting and differentiating into multiple tissue types. However, recent studies have revealed that implanted cells do not survive for long, and that the benefits of MSC therapy could be due to the vast array of bioactive factors they produce, which play an important role in the regulation of key biologic processes. Secretome derivatives, such as conditioned media or exosomes, may present considerable advantages over cells for manufacturing, storage, handling, product shelf life and their potential as a ready-to-go biologic product. Nevertheless, regulatory requirements for manufacturing and quality control will be necessary to establish the safety and efficacy profile of these products. Among MSCs, human uterine cervical stem cells (hUCESCs) may be a good candidate for obtaining secretome-derived products. hUCESCs are obtained by Pap cervical smear, which is a less invasive and painful method than those used for obtaining other MSCs . Moreover, due to easy isolation and a high proliferative rate, it is possible to obtain large amounts of hUCESCs or secretome-derived products for research and clinical use. Stem cells are immature tissue precursor cells that are able to self-renew, have the ability to form clonal cell populations, and differentiate into multiple cell lineages ,2. TheseThe therapeutic potential of stem cells can be attributed to three key mechanisms of action. The first is “homing”, for which systemically administered stem cells migrate to the focus of acute injury due to chemical gradients. It is hypothesized that migration to target tissues occurs through a process similar to that of leukocyte migration. Chemoattraction is mediated by cell surface receptors such as the chemokine receptors. Although the exact mechanism of stem cell-endothelial interaction at the target site is not well established, integrins and selectins mediate these interactions ,4. TransAdult stem cells guarantee the maintenance and repair of adult tissues and organs. Mesenchymal stem cells (MSCs) are especially emerging as cell-based therapy of numerous diseases.Adult bone marrow MSCs (BMMSCs) were discovered in 1968 by Friedenstein et al. , skin 11, lung , synoviaAlthough MSCs are present in multiple tissues, their overall quantity in the body is scarce. Cell therapy protocols generally require hundreds of millions of MSCs per treatment; therefore, cell expansion in vitro is needed for about 10 weeks before implantation. The patient’s age and clinical characteristics influence the optimal culture conditions for clinical scale production of human MSCs ,31. The Earlier research attributed the therapeutic effects of MSCs to their engrafting and differentiation capacity. However, several studies have revealed that the implantation time of MSCs is usually too short to have an effective impact ,35,36,37We are presently witnessing the emergence of a novel type of biological regulation involving the communication between cells via their secreted substances, the secretome. The secretome is defined as the set of factors/molecules secreted to the extracellular space. These factors include, among others, soluble proteins, free nucleic acids, lipids and extracellular vesicles. The latter can be subdivided into apoptotic bodies, microparticles and exosomes . The secThe use of cell-free therapies such as MSC-sourced secretome in regenerative medicine provides key advantages over stem-cell based applications: (a) application of the secretome resolves several safety considerations potentially associated with the transplantation of living and proliferative cell populations including immune compatibility, tumorigenicity, emboli formation and the transmission of infections; (b) MSC-sourced secretome may be evaluated for safety, dosage and potency in a manner analogous to conventional pharmaceutical agents; (c) storage can be done without application of potentially toxic cryopreservative agents for a long period without loss of product potency ,49,50; . The most promising potential applications involve the treatment of graft versus host disease -5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1), an NF-κB inhibitor, can attenuate the secretion by MSCs of pro-inflammatory factors for at least six days in vitro. CM derived from TPCA-1-loaded MSCs also showed reduced ability to attract human monocytes and prevented differentiation of human cardiac fibroblasts to myofibroblasts . This isMSCs have drawn much interest for their therapeutic benefits in immune modulation and tissue remodeling. Earlier research primarily attributed this to the ability of transplanted MSCs to engraft and to differentiate into multiple tissue types. However, there are several limitations that currently impair the wide widespread use of MSCs. Some studies suggest that the transportation of MSCs into normal tissues might cause tumor formation. Recent studies have revealed that implanted MSCs do not survive for long, suggesting that the benefits of MSC therapy might be attributable to their secreted factors. An expanding body of recent literature has brought attention to the vast array of bioactive factors produced by MSCs including cytokines, growth factors, microRNAs, proteasomes, and exosomes, which may play an important role in the regulation of numerous physiological processes. Secretome-based approaches using CM or exosomes may present considerable potential advantages over living cells in terms of manufacturing, storage, handling, product shelf life and their potential as a ready-to-go biological therapeutic agent. Multiple experimental studies demonstrate that secretome-derived products are sufficient to significantly improve multiple biomarkers of pathophysiology in many animal models of different diseases. However, composition and effects of secretomes differ based on MSC anatomical origin. Using hUCESCs, in particular, presents important advantages. hUCESCs are obtained by Pap cervical smear, which is a less invasive and painful method than those used for obtaining other MSCs . In addition, due to their easy isolation and high proliferative rate, it is possible to quickly obtain large amounts of hUCESCs or secretome-derived products for research and clinical use.Considering that multiple clinical trials involving MSCs have been approved by national agencies, it is perfectly reasonable to expect approval of secretome-derived products from MSCs. Although the composition of secretome-derived products from MSCs is complex, this in itself should not be an impediment for regulatory approval of a regenerative product. For example, platelet-rich plasma or amniotic fluid, which are highly complex and include numerous growth factors and exosomes that remain poorly characterized, are routinely used as a regenerative therapy for multiple applications in wound healing and orthopedics. Exosomes sourced from dendritic cells have already reached the clinical-trial stage for immunotherapy of certain cancers. Nevertheless, regulatory requirements for manufacturing and quality control will be necessary to establish the safety and efficacy profile of these products.