Diabetes highlights a growing epidemic imposing serious social economic crisis to the countries around the globe. Despite scientific breakthroughs, better healthcare facilities, and improved literacy rate, the disease continues to burden several sections, especially middle and low income countries. The present trends indicate the rise in premature death, posing a major threat to global development. Scientific and technological advances have witnessed the development of newer generation of drugs like sulphonylureas, biguanides, alpha glucosidase inhibitors, and thiazolidinediones with significant efficacy in reducing hyperglycemia. Recent approaches in drug discovery have contributed to the development of new class of therapeutics like Incretin mimetics, Amylin analogues, GIP analogs, Peroxisome proliferator activated receptors, and dipeptidyl peptidase-4 inhibitor as targets for potential drugs in diabetes treatment. Subsequently, the identification and clinical investigation of bioactive substances from plants have revolutionized the research on drug discovery and lead identification for diabetes management. With a focus on the emerging trends, the review article explores the current statistical prevalence of the disease, discussing the benefits and limitations of the commercially available drugs. Additionally, the critical areas in clinical diabetology are discussed, with respect to prospects of statins, nanotechnology, and stem cell technology as next generation therapeutics and why the herbal formulations are consistently popular choice for diabetes medication and management. Diabetes is a major killer worldwide and its unprecedented rise poses a serious threat to mankind. According to recent estimation, 387 million people worldwide are affected from the disease with a prevalence rate of 8.3% and 46.3% still remains undiagnosed . Furtherβ cell defects, dipeptidyl peptidase-4 (DPP-4) inhibition by Sitagliptin, and increased islet survival [The selection and application of a glucose lowering therapy are dependent on a number of considerations like the severity of hyperglycemia, hepatic and renal associated functions, risks of hypoglycemia, body mass index, ability to self monitor the blood glucose level, and also the cost of the medication. The therapeutics for type 1 diabetes includes stimulation of insulin secretion through GLP analogues like Exenatide and Liraglutide , 9, insusurvival , 11 and survival .The treatment approach for type 2 diabetes includes several conventional therapeutics, namely, sulfonylureas and repaglinide enhance insulin secretion, troglitazone increases insulin action in fat and muscle, metformin promotes insulin mechanism in liver tissue, and miglitol and acarbose enact delayed carbohydrate absorption from food intake, respectively . The druSecondary Forms of Diabetes. Subsequently, secondary forms of the disease may arise due to defects/mutations in genome of the organism: gene mutations in MODY1-hepatocyte nuclear factor-4-alpha (HNF4α) in chromosome 20q12-q13.1; MODY2-glucokinase (GCK) gene mutation in chromosome 7p15-p13; MODY3-hepatocyte nuclear factor-1-alpha (HNF1A) gene mutation in chromosome 12q24.2; MODY4-insulin promoter factor-1 (IPF1) gene mutation in chromosome 13q12.1; MODY5-hepatic transcription factor-2 (TCF2) gene mutation, chromosome 17cen-q21.3; MODY6- neurogenic differentiation 1 (NEUROD1) gene mutation in chromosome 2q32; MODY7-Kruppel-like factor 11 (KLF11) gene mutation in chromosome 2p25; MODY8 (or diabetes-pancreatic exocrine dysfunction syndrome)-carboxylester lipase (CEL) gene mutation, chromosome 9q34; MODY9-paired box gene 4 (PAX4) mutation, chromosome 7q32:Mutations occurring in mitochondrial genome, referred to as mitochondrial diabetes MELAS syndrome .Genetic syndromes, Klinefelter's syndrome, Turner's syndrome, Down syndrome, Prader-Willi, Laurence-Moon-Biedl, Friedreich's ataxia, Huntington's chorea, myotonic dystrophy, and porphyria.Drug/chemical induced diabetes, Thiazides, DPH, α-interferon, L-asparaginase, vacor, nicotinic acid, pentamidine, steroids, levothyroxine, and diazoxide [iazoxide .However, the disease together with its multiple complications puts forward the immediate requirement to act with a well defined strategy. The underlying platform is aimed at achievement of complete glycemic regulation, possible through assessment of present glycemic status, and analysis of the associated disorder would aim at allocating the healthcare facilities to the affected people . The newThe present trends in diabetes therapeutics and management have highlighted an urgent requirement for extensive investigations aiming at identification and clinical trials of natural products and their analogues in drug discovery studies. In an attempt to address the global issue, the paper provides a comprehensive update highlighting the global scenario and statistical prevalence of diabetes. Furthermore, the emerging trends in clinical diabetology were discussed, exploring the advantages as well as the limitations of the commercially available therapeutics. The present era has witnessed the development of next generation therapeutics, statins, nanotechnology, and stem cell technology and the reasons why the natural products and analogues define a prospective field in diabetes medication and management.On the basis of insulin deficiency, diabetes can be classified into the following types as follows.β cell destruction varies showing fast deterioration in infants and children while the degeneration of β cells is slower in adults. Symptoms like ketoacidosis occur in children and young individuals while others exhibit modest fasting hyperglycemia that can change to severe hyperglycemia or ketoacidosis in response to stress or infection [It is also known as juvenile onset diabetes or type 1 diabetes, which accounts for 5–10% of the patients, resulting from cellular-mediated autoimmune destruction of the pancreatic cells. The disease can affect people of all ages but usually occurs in children or young adults. Regular supply of insulin injections is essential for the control of glucose level in blood. The rate of nfection . These pnfection . This fonfection .  Table A minor number of patients having type 1 diabetes, mostly of Asian and African ancestry, have no etiologies. These are prone to ketoacidosis and have permanent insulinopenia. The occurrence of ketoacidosis is in episodes and the level of insulin deficiency between episodes fluctuates. Idiopathic diabetes has genetic predisposition and an absolute need for insulin replacement therapy depends on the condition of the patient .It is also referred to as adult onset diabetes, which accounts for 90–95% of all diabetes. Major metabolic syndromes like obesity, insulin resistance, and dyslipidaemia have led to an epidemic of type 2 diabetes . The treIt is defined as any degree of glucose intolerance resulting in hyperglycaemia of variable severity that is diagnosed during pregnancy . GDM, orDiabetic ketoacidosis (DKA) is a condition, arising due to infection, inadequate insulin or poor insulin compliance, acute pancreatitis, stroke, drugs, metabolic disturbances within the body, or negligence with the treatment . The uncThe interface of nanotechnology in the treatment of diabetes has introduced novel strategies for glucose measurement and insulin delivery. Researchers have demonstrated the advantages of glucose sensors and closed-loop insulin delivery approaches in facilitating the diabetes treatment to make it  beneficiA nanomedical device is a microcapsule containing pores which has been a promising tool in the drug delivery approach. These pores are considerably large to allow the passage of small molecules such as oxygen, glucose, and insulin but are small enough to allow the movement of larger immune system molecules such as immunoglobulins and graft-borne virus particles. Microcapsules containing replacement islets of Langerhans cells, mostly derived from pigs, could be implanted beneath the skin of diabetes patients. This could temporarily restore the body's delicate glucose control feedback loop without the need for powerful immunosuppressants that can leave the patient at serious risk for infection  Table 2 The nanoparticle targeted drug delivery approach has enormous benefits which include the improved bioavailability of drugs by targeting specific tissues, organs, and tumors thereby providing the highest dose of drug directly at the targeted site. One of the biggest technological challenges is the scalability of a nanoparticle. Manufacturing three-dimensional nanostructures as compared to stand-alone or two-dimensional layer-shaped nanosurfaces is a complex task since manufacturing techniques are yet to be standardized. Another apprehension is that the exposure to nanoparticles might be toxic or hazardous. Concerns about the potential ill effects of engineered nanomaterials such as carbon buckyballs and nanotubes through inhalation, ingestion, or absorption through the skin are increasing .Insulin forms an essential requirement for type 1 and type 2 advanced diabetes and the traditional systems of insulin delivery included infections, painful administration, and poor compliance of patients. However, recent micro- and nanotechnologies have facilitated the insulin administration process through regulation of insulin delivery constituting pulmonary, nasal, transdermal, and closed-loop delivery .Statins are defined as inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A and inhibit the crucial process of LDL cholesterol in liver, thereby decreasing its level in the blood besides increasing healthy blood vessel lining . Since tStatin therapy reduces low density lipoprotein (LDL) cholesterol to a significant level thereby greatly decreasing the chances of developing a coronary artery disease . NationaStatins have good efficacy and are effective in lowering cardiovascular events in people with modest levels of cholesterol and without cardiovascular disease. However, the HMG-CoA reductase inhibitors or statin therapy also has some disadvantages. The therapy has some side effects like renal dysfunction and muscle disorders from myositis to frank rhabdomyolysis and hepatic dysfunction which is rare and can be tolerated by the patient . The triβ cell deficiency of the pancreatic cells, resulting in insufficient insulin secretion. The strategies should aim at either removing the defects in pancreatic β cell or enhancing the sensitivity of the body cells to the action of insulin. β cell replacement strategies offer a novel source while current strategies aiming at islet cells and pancreas transplantation are limited due to shortage of donor organs [The interest to find a possible therapeutic for diabetes has eventually explored various new scientific areas of research, with the stem cell technology being one of them. It is known that both type 1 and type 2 diabetes result from the r organs .β cells, type 2 diabetes results from irregularities in β cells function together with insulin resistance in peripheral organs [ in vitro and in vivo conditions due to direct contact and production of soluble markers [β cell function in newly diagnosed type 1 diabetic patients [In contrast to type 1 diabetes, which is caused by autoimmune destruction of pancreatic l organs . Mesench markers –49. MSCspatients . Furtherpatients . in vitro and in vivo [Some studies have shown that bone marrow derived MSCs have the ability to differentiate into insulin producing cells both in vivo –54. The  in vivo . Further in vivo . Still,  ex vivo gene therapy is described as the one in which the tissues are removed from the body; the therapeutic gene is inserted in vitro and then reimplanted back in the body while the in vivo therapy involves the insertion of gene therapy vectors directly to the patients by subcutaneous, intravenous, or intrabronchial routes, or by local injection [ ex vivo therapy aims at the generation of cells which possess the properties of β cells, for example, insulin producing cells [β cells for transplantation. However, the concern lies in the aspect of surgically removing the tissue from the patient and reimplantation of the genetically modified tissues back into the body of the patients [β cells and islet transplantation has been explored as a possible solution for the treatment. The invention of insulin gene therapy substitutes β cell function by generating insulin secretory non-β cells, not vulnerable to autoimmune reactions, offering a prospective therapeutic approach for type 1 diabetes [The series of experiments leading to cloning and expression of insulin in the cultures cells in the 1970s was a tremendous revolution in the field of medicine and application of gene therapy in the treatment of diabetes was suggested as a possible cure. Regulating the sugar levels is the most important aspect in the treatment which also reduces the complications associated with the disease. Somatic gene therapy involving the somatic cells of the body includes two methods of gene delivery. The first one known asnjection . The appng cells . This thpatients . Furtherdiabetes . in vivo gene therapy is the method of choice as a therapeutic strategy because it is simpler and the vector containing the desired gene is directly inserted into the patient, but the development of safe (not toxic to host) and effective vectors remains as a challenging task for gene therapist. Presently, the strategies for in vivo therapy involve three methods: genetic transfer of glucose lowering genes which are noninsulin in nature. Presently, the strategies for in vivo therapy include genetic transfer of glucose lowering genes which are non-insulin in nature and application of blood sugar lowering genes: an enhancer of glucose utilization by liver or skeletal muscles and an inhibitor of glucose production by the liver [ Gck enhances glucose utilization by the body [Thehe liver . For exahe liver . It was the body . The genthe body . In anotthe body , 64. Thethe body .β cells production in the liver [Other areas of genetic engineering include transfer of genes which show response to glucose and the use of gene therapy to induce he liver . The gluhe liver , 66. Thehe liver –69. The he liver  having 2he liver .β cells formation in the liver. Kojima et al. reported that it is possible to induce the formation of β cells by the endocrine cells by delivering islets specific transcription factors [β cells neogenesis seems to be a promising approach as a therapeutic for diabetes, since it can offer a solution for the autoimmunity in type 1 diabetes [Research has also been carried out to induce the synthesis of  factors , 73. The factors . The strdiabetes .Medical nutrition therapy in prevention and management of diabetes puts forth numerous advances in clinical research, aiming to use nutrition therapy for the treatment of disorders and illnesses. American Diabetes Association in 1994 coined the term “medical nutrition therapy” constituting 2 phases, namely, adjudging the nutritional requirement of a person and treatment through counseling and nutrition therapy, respectively . The obj Allium sativum Linn. (Liliaceae), Gymnema sylvestre (Retz.) Schult (Asclepiadaceae), Murraya koenigii (L.) Spreng. (Rutaceae), Allium cepa (Liliaceae), Withania somnifera Dunal (Solanaceae), and Ferula foetida Linn. (Umbelliferae) have been found to possess antidiabetic properties when assessed in experimental models of diabetes. The antidiabetic properties of G. sylvestre had been discussed in detail [Literature has suggested the utilization of herbal medications for the treatment of insulin dependent and noninsulin dependent diabetes since time immemorial. Plants possessing antidiabetic properties may be suitable as adjunct to the existing therapies or as a prospective source of new hypoglycemic compounds. Since time immemorial, naturopathic therapies have been applied for a number of health ailments and continue to gain popularity in the present arena as well. Ancient literature revealed that diabetes was a known disease since Brahmic period and finds a mention in Ayurvedic literature, Sushruta samhita written in fourth and fifth centuries BC . Two forn detail , 85 owinDiabetes has remained as one of the most challenging health problems in the 21st century accounting for a global presence. Diabetes is a serious public health problem, but the good news is that important advances are being made in prevention, detection, and treatment of diabetes. For the management of type 1 diabetes, patients require insulin administration 3-4 times a day throughout their lives and their blood sugar levels should be regularly monitored to avoid complications like retinopathy and risks of cardiovascular diseases. It has been estimated that around 1300 patients with type 1 diabetes receive whole organ (pancreas) transplant and do not require insulin infusion but the demand for organs transplantation is higher than supply. Another risk factor is the rejection of transplanted organ; therefore, patient is given strong immunosuppressive drugs which can lead to other serious diseases .β cell function is essential since it can lead to a loss of glycemic control. Conventional drugs and insulin are effective but cannot repair the associated metabolic and glucoregulatory dysfunctions. The menace of diabetes is increasing day by day and aggressive and targeted combinational therapy is the need of the hour particularly incretin based therapy and peptide analogs. This may restore and preserve β cell function and halt the progression of type 2 diabetes [For the management of type 2 diabetes, a well monitored glycemic control is required. The need to control the progressive deterioration of diabetes . In the The prospects of leptin therapy are one of the emerging trends in the treatment of diabetes. It is a hormone secreted by adipocytes, which acts on the neurons within the central nervous system. The multiple actions of this hormone include control of excessive increase in weight, by suppressing the intake of food and increasing the expenditure of energy . Leptinshttp://www.ondrugdelivery.com/, 2006). A research by Odegaard and colleagues revealed that activated macrophages display a beneficial role in the regulation of nutritional homeostasis and suggests that polarization of the macrophages towards the alternative state might be a useful possibility in the treatment of type 2 diabetes [Another area of drug research includes designing and use of mucoadhesive microcapsules of various drugs like glipizide to achieve controlled release of the drug and its effective targeting. Mucoadhesion has been a novel approach in drug delivery designing because it causes the slow release of the drug at the action or absorption site thereby enhancing the interaction of the drug with the underlying tissue forms, enhancing the bioavailability of the drugs . There idiabetes .Great strides have been made clinically in the prevention, development, and treatment of the disease but no therapeutic method have been completely successful till date. With new technologies revolutionizing the treatment possibilities, the search for an effective medication is not far ahead. The extensive research leading to the discovery of the pathway genes contributing to the development of the disease and the sequencing of complete genomes have revolutionized the diabetes research. The development of the techniques like the PCRs, DNA microarray, and gene knockouts with silencing has opened up a new area in the identification of the defective genes/mutations in the genome of the organism. The increasing prevalence of diabetes globally is creating a financial burden on the economy of the respective country. Unlike some other diseases, treatment exists for diabetes, and if managed correctly, it is very effective in reducing complications such as heart attacks, amputations, blindness, and kidney failure. With the ongoing research, a right therapeutic for the treatment of diabetes is not unachievable.


Pulmonary hypertension complicating idiopathic pulmonary fibrosis, also known as secondary pulmonary hypertension, represents a major source of morbidity and mortality in affected patients. While the study of primary pulmonary arterial hypertension has yielded several therapies, the same is not true for the treatment of pulmonary hypertension secondary to pulmonary fibrosis. Recent studies have indicated an important role of hypoxia-inducible factor (HIF) – a regulatory protein that is vital in adaptation to hypoxic conditions – in the development of secondary pulmonary hypertension. HIF influences development of hypoxia-induced pulmonary hypertension through alteration in voltage-gated potassium channels and homeostatic calcium regulation, resulting in disruption of endothelial cell-cell communication, and eventual vascular remodeling. This article summarizes salient literature related to HIF and secondary pulmonary hypertension, in addition to proposing a final common pathway in known mechanistic pathways that result in endothelial barrier integrity loss – vascular “leak” – primarily through a shared endothelial-epithelial signaling protein family, CCN. Unfortunately, the available medical therapies for treatment of primary pulmonary arterial hypertension are either ineffective in the treatment of secondary pulmonary hypertension , or provide only a small potential benefit in secondary trial outcomes . To this end, there is a large need for novel therapies based on alternative mechanistic pathways in disease development.Idiopathic pulmonary fibrosis (IPF) is a progressive parenchymal lung disease without significant disease-modifying treatment. Survival time after diagnosis is widely variable and depends on a variety of factors including age, pulmonary function testing and degree of fibrosis. One additional factor contributing to death associated with IPF is the development of secondary pulmonary hypertension, defined as WHO group III pulmonary hypertension – associated with lung diseases and/or hypoxia. Prevalence of secondary pulmonary hypertension in IPF ranges from 32% to 85%, and importantly is associated with worse exercise capacity, quality of life and survival compared to patients with IPF and no evidence of pulmonary hypertension , 5. While the literature on the topic is sparse, at least one murine study utilizing a model of pulmonary fibrosis – adenoviral delivery of TGF-β1 – showed that administration of a downstream hypoxia-response element target protein (VEGF) ameliorated pulmonary hypertension, yet may have exacerbated the fibrotic response .One such candidate pathway is that of hypoxic signaling in the pathogenesis of secondary pulmonary hypertension, specifically the role of hypoxia-inducible factor (HIF). HIF is a key regulator of cellular adaptation to hypoxia, leading to increased transcription of hypoxia-response element genes, such as vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF). Existing in nature as a heterodimer with an α- and β-subunit, two isoforms of HIF (HIF1α and HIF2α) have been shown to influence development of secondary pulmonary hypertension in a chronic hypoxia model of secondary pulmonary hypertension .The purpose of this article is to both explore current models of how HIF influences development of secondary pulmonary hypertension, and to examine potential future directions in research and treatment of this prevalent – conservatively, up to 4 per 100,000 people in the United States – and devastating illness . It has also been known for some time that hypoxia acts directly on the pulmonary artery smooth muscle to alter Ca2+ transport and increase contractility . More recently, in a model of chronically hypoxic rats, endothelin-1 (ET-1) – a potent vasoactive peptide in pulmonary vasculature – was demonstrated to increase the calcium sensitivity of pulmonary artery smooth muscle cells, suggesting a shared downstream mediator with pulmonary arterial hypertension .Examining the effect of calcium antagonists on hypoxic pulmonary vasoconstriction, it is known that the hypoxic mechanism is critically dependent on the transmembrane influx of calcium 2 and treated with either a HIF-1 translational inhibitor or vehicle. They were then able to demonstrate that not only did HIF-inhibition blunt the increase in right ventricular pressure and remodeling, it did so – in part – by decreasing the amount of intracellular calcium within pulmonary artery smooth muscle cells . This was independent of whether HIF-inhibition occurred prior to or after development of hypoxia-induced pulmonary hypertension.Investigators from The Johns Hopkins University have been integral in establishing a link between HIF and calcium channel homeostasis in the development of secondary pulmonary hypertension. In one study, the group exposed mice to three weeks of 10% FiO, suggesting a potential mechanism for a chronic HIF-controlled change in vascular remodeling.While tonic hypoxic vasoconstriction almost certainly plays a role in the long-term remodeling of the pulmonary vasculature, it does not by itself establish a link with the late manifestations of disease demonstrating perivascular fibrosis and vessel thickening. To this end, in a second related study, the same group established a decrease in pulmonary artery smooth muscle cell migration with inhibition of hypoxia-induced, and calcium channel-mediated, development of pulmonary hypertension . A key determinant of intracellular calcium concentration in the pulmonary artery smooth muscle cell is the resting membrane potential, as this influences voltage dependent sarcolemmal Ca2+ channels. The resting membrane potential is in turn tightly regulated by voltage-gated potassium channels. These channels are known to have impaired function within the cell membrane upon exposure to chronic hypoxia. In a study exposing isolated rat intrapulmonary arterial cells to greater than 24 hours of hypoxia, Wang and colleagues  showed that the decrease in KV channel subtypes 1.2 and 1.5 was a transcriptionally regulated event, data that were later confirmed in an in vivo model of subacute hypoxic exposure with subsequent development of secondary pulmonary hypertension . The phenotype is rescued with gene therapy targeted to increase KV1.5 channel expression, with decrease in hypoxic vasoconstriction and resolution of pulmonary hypertension .A novel potassium channelopathy, KCNK3, was recently linked to the development of familial pulmonary hypertension, suggesting a potential role of potassium channels in the development of secondary pulmonary hypertension V channel expression was later found to be at least partially HIF-mediated by Bonnet and colleagues. In their study, a rodent model of pulmonary hypertension with activation of HIF1α had inhibited expression of voltage-gated channel, KV1.5 . Blocking HIF activation decreased pulmonary vasoconstriction and downstream vascular remodeling, suggesting that manipulation of the protein pathway may lead to further pharmaceutical targets of disease.The link between hypoxia exposure and subsequent alteration in K. Using a model of pulmonary hypertension associated with pulmonary fibrosis, our group has recently demonstrated that mice with BMPRII mutation expressed a high amount of HIF1α in the lung, corresponding with worsening pulmonary hypertension . Thus, vascular “leak”, related to hypoxic signaling, may play a role in the progression of pulmonary hypertension. We also know that calcium entry into the endothelium influences barrier integrity, leading to immediate and delayed clinical consequences such as alveolar flooding and impairment of gas exchange . Prior work by Irwin and colleagues  demonstrated that not only does hypoxic-exposure in cultured human pulmonary artery endothelial cells cause increased albumin permeability and HIF1α accumulation, but treatment of hypoxia-exposed cells with HIF1α siRNA lowers monolayer permeability. A similarly designed study examining hypoxic exposure in an in vivo murine model found the same results, with downstream inhibition of HIF-dependent molecules resulting in protection against extravasation of Evans blue dye, indicating preserved barrier integrity .A clue linking the mechanism of the above derangements with the subsequent development of secondary pulmonary hypertension can be found in the examination of BMPRII-related pulmonary arterial hypertension. It has previously been shown that BMP receptor signaling controls vascular remodeling during angiogenesis by maintaining the expression of endothelial regulatory molecules, with the attenuation of the signal resulting in predisposition to compromised vascular integrity , and therefore represent an attractive area of research in the treatment of secondary pulmonary hypertension. CCN proteins are tightly regulated by HIF under hypoxic conditions involved in a diverse array of tissue beds and functions, such as appropriate placental invasion in trophoblast cell lines (Cyr61 and NOV) , chondrocyte development and survival (CTGF)  and gastric cancer cell invasion (Cyr61) . In addition CCN-4, also known as WISP-1, has been shown to be necessary in the development of alveolar epithelial cell hyperplasia and epithelial-to-mesenchymal transition (EMT) leading to pulmonary fibrosis . Such a unique array of functions provides an interesting intersection between epithelial and endothelial cell biology, making the CCN pathway a very attractive target for future research , 29. In support of this assertion, our lab has recently shown that through deletion of HIF signaling within the vascular endothelium of mice, pulmonary hypertension is normalized in both the pulmonary fibrosis and chronic hypoxia models of disease . We furthermore demonstrated a correlative relationship with decreased CTGF expression, an alteration in vascular permeability, and degree of vascular remodeling. These findings support further exploration into tissue-specific targeting of the CCN family of proteins, including CTGF, for pharmacologic manipulation of the hypoxia-signaling pathway.Consequently, targeting a HIF-mediated molecular pathway that regulates both intracellular calcium and endothelial cell communication – influencing vascular integrity – would logically be of potential therapeutic benefit. The CCN family  are one such group of regulatory proteins that are involved in both endothelial cellular signaling and intracellular calcium trafficking While much is known from existing literature on the development of secondary, mainly hypoxia-induced, pulmonary hypertension, this knowledge base has afforded little in the way of directed therapy. The mechanism that HIF plays in the development and progression of disease has provided intriguing insight into the future of treatment. Working on the background of established pathways, including the role of HIF in potassium-channel mediated cell membrane potential and calcium homeostasis, a new direction has been established for examining shared pathways in disruption of the barrier integrity of the lung. Pursuing research along these lines, with the CCN family of proteins being a primary candidate for study, will provide the impetus to follow Benjamin Franklin’s timeless advice, and avoid allowing a “small leak” to continue to plague our efforts at preserving our pulmonary vessel, a “great ship.”


P<0.1) directly interact more than random expectation and are involved in a limited number of interconnected biological processes, indicating that they are functionally related. The functionally coherent networks generated by this approach contain ASD-relevant disease biology, as demonstrated by an improved positive predictive value and sensitivity in retrieving known ASD candidate genes relative to the top associated genes from either GWAS, as well as a higher gene overlap between the two ASD datasets. Analysis of the intersection between the networks obtained from the two ASD GWAS and six unrelated disease datasets identified fourteen genes exclusively present in the ASD networks. These are mostly novel genes involved in abnormal nervous system phenotypes in animal models, and in fundamental biological processes previously implicated in ASD, such as axon guidance, cell adhesion or cytoskeleton organization. Overall, our results highlighted novel susceptibility genes previously hidden within GWAS statistical “noise” that warrant further analysis for causal variants.Genome-wide association studies (GWAS) for Autism Spectrum Disorder (ASD) thus far met limited success in the identification of common risk variants, consistent with the notion that variants with small individual effects cannot be detected individually in single SNP analysis. To further capture disease risk gene information from ASD association studies, we applied a network-based strategy to the Autism Genome Project (AGP) and the Autism Genetics Resource Exchange GWAS datasets, combining family-based association data with Human Protein-Protein interaction (PPI) data. Our analysis showed that autism-associated proteins at higher than conventional levels of significance ( Autism Spectrum Disorder (ASD) is a complex neurodevelopmental illness with significant clinical and genetic heterogeneity. Family and twin studies demonstrated that ASD is one of the most heritable neuropsychiatric disorders, but there is yet no consensus on the underlying genetic architecture loci to remain hidden within the GWAS statistical “noise”. Evidence from classical quantitative genetic analysis further suggests that most of the heritability missing in complex diseases is rather hidden below the threshold for genome-wide significant associations Devlin et al. (2011) have recently predicted that common variants having an odds ratio of 1.5 or more are very unlikely to exist; few, if any, common variants with an impact on risk exceeding 1.2 may still await discovery, but require much larger sample sizes, while variants with modest impact may range from zero to many thousands a priori sorting the genes into incompletely annotated predefined gene sets, is emerging as an appealing unbiased alternative to pathway analysis. Network-based approaches have been widely applied in the analysis of high-throughput expression data from a wide range of diseases loci in human GWAS data New strategies are therefore needed to increase the power of GWAS analysis. The use of molecular networks, which is not limited by Based on the premise that disease-causing genes are likely to be functionally related, in the present study we applied a network-based approach to two ASD GWAS datasets, the AGP consortium GWAS and the GWAS carried out in the Autism Genetic Resource Exchange (AGRE) dataset A workflow of the strategy for network definition, validation and identification of the most relevant candidate genes is shown in All the data used is previously published and publicly available. Written informed consent has been previously obtained from all families and procedures had approval from institutional review boards from all the institutions involved in recruitment and research, following national and international ethical and legal regulations and the principles of the Declaration of Helsinki.The AGP dataset included 2818 trios consisting of autistic patients and both parents collected as part of the AGP Consortium. Patients were diagnosed and genotyped as previously reported The GWAS replication dataset from the Autism Genetic Resource Exchange (AGRE) included 943 ASD families  from the AGRE cohort Summary SNP association results were obtained from the database of Genotype and Phenotype (dbGAP) repository for 6 case-control GWAS for other pathologies, including Parkinson's Disease (PD) P-value among the TDT P-values of all individual SNPs mapped to that gene. MAGENTA then uses step-wise multivariate linear regression analysis to regress out of this P-value the confounding effects of gene size, number of SNPs per kilobase (kb), number of independent SNPs, number of recombination hotspots and the number of linkage disequilibrium units per kb.Genotyped SNPs from the AGP and AGRE GWAS were assigned to specific genes if they were located within or up to 10 kb from the gene, using the GRCh37/hg19 genome build (Step 1). Each gene was assigned a gene score using MAGENTA  P-value cutoffs (0.5<-LogP<5) were superimposed onto their corresponding protein on a large human protein-protein interaction (PPI) network, converting Entrez gene IDs to Uniprot IDs (release 2010_04) (Step 2). This PPI network, covering 12372 proteins and 58365 interactions, was previously built compacting data from six public PPI databases: BIND, BioGRID, HPRD, IntAct, MINT and MPPI Genes selected at various gene-wise P-value for which corresponding proteins were functionally connected beyond random expectation, thus the lowest gene-wise P-value for which there is still relevant biological data in the GWAS that can be captured through network analysis (step 3). Three metrics indicative of this functional coherence were estimated for various association gene-wise P-value thresholds, for the two ASD datasets, and compared with those determined for 1000 equal size sets of randomly selected proteins from the human PPI network. The metrics evaluated were 1) the percentage of proteins directly interacting; 2) the percentage of isolated nodes, which represents the fraction of selected proteins with no interactions with any other selected protein; and 3) the size of the largest connected component (LCC), the largest group of selected proteins that are reachable from each other in the network. An empirical P-value was obtained computing the fraction of random samples where the value of the network metric is greater  than the observed one. Network analysis was performed using python module Network X.Topological properties from the resulting network were analyzed to select the gene-wise https://gene.sfari.org/autdb/Welcome.do).To evaluate the performance of the proteins included in the LCC in retrieving known ASD candidate genes, the precision and recall against a curated list of ASD candidate genes were calculated (step 4). This list was obtained from SFARIGene and includes 236 genes having at least minimal evidence of association with ASD (categories 1 to 4) or categorized as syndromic  the same number of top genes  as those included in the LCC.Precision  is the proportion of known candidate genes among the selected genes, while recall (Sensitivity) is the proportion of known candidate genes retrieved by the selection. The precision and recall calculated for the genes encoding LCC proteins were compared to those determined using two other gene selection criteria: a) all genes selected at the same gene P-value) and top genes .Overlap between the AGP and AGRE datasets at SNP, gene and LCC levels was determined using the Jaccard index, defined as the size of the intersection divided by the size of the union of the datasets. For comparison purposes the size of each dataset LCC was used to select from each GWAS dataset an equal number of top SNPs . Each protein included in the AGP LCC had an initial score of 0.5. If the protein was present in the AGRE ASD dataset LCC, 0.5 was added to the initial protein score, whereas for each unrelated disease dataset LCC where the protein was present, one sixth of 0.5 was subtracted from the score. Therefore, protein scores vary between 0 and 1, with zero representing a protein present in the LCCs of the AGP dataset and the 6 unrelated diseases, while a score of 1 is attributed to a protein present only in the LCCs of both ASD datasets.th; http://david.abcc.ncifcrf.gov) P-values were corrected by the Benjamini correction. Top-scoring genes were further investigated using NextBio platform , a curated and correlated repository of experimental data derived from an extensive set of public resources (eg. ArrayExpress and GEO) Functional enrichment was tested by DAVID , followed by the integration of GWAS data onto protein interaction data. Then, we determined the lowest gene-wise P-value threshold for which genes encoding the network proteins were functionally related, inferred by their proximity in the network. Statistical noise is expected to have random connections in the network, while disease proteins are more likely to establish direct interactions between them and more rarely be isolated in the network, translating into a larger group of proteins that are all interconnected. For both ASD datasets, proteins encoded by genes selected at a gene-wise -Log10P cutoff between 0.5 and 1.5 were found to establish significantly more direct interactions than equal sized sets of randomly selected proteins , with the significance maintained up to -Log10P = −2.0 in the case of AGRE dataset , again with significant differences maintained for lower gene-wise P-values in the AGRE dataset   . The larP-value for which the percentage of direct interactions was significantly higher, the percentage of isolated nodes significantly smaller and the size of the LCC significantly larger than random expectation (in AGP dataset), but additionally capture low effect genes, such as TSC2 (Uniprot P49815), which single gene association analysis alone does not have the power to detect.Concerning the proportion of known genes that are retrieved by our selection, or recall, LCC encoding genes had a lower recall compared with all genes selected at the same cutoff, as expected since LCC genes are a subset of this selection . HoweverTBK1 gene) and 10 genes in common between the two datasets, when comparing the same number of SNPs or genes  than genes included in the LCCs from each dataset. Remarkably, we observed a 25 and 2.5-fold increase in the overlap between the two ASD datasets (AGP and AGRE) at PPI network level when compared to SNP or gene level, respectively . The largest connected components generated by genes selected at -Logde novo mutations in ASD described in at least one of the three whole exome sequencing studies recently published CSDE1 (Uniprot O75534), PGD (Uniprot P52209), TSC2). In addition, 80 (∼19%) of the AGP LCC-encoding genes were deleted or duplicated by CNVs identified by the AGP whole genome analysis as potentially pathogenic (with less than 50% of length overlap with control datasets) (A closer inspection of these LCCs at the gene level showed that around 30 (7–8%) of the encoding genes were implicated in neuropsychiatric or neurodegenerative disorders . More inatasets) .To further examine the specificity of the proteins in the AGP LCC for ASD, this network was compared with LCCs generated from six unrelated diseases GWAS . Based on the presence of each protein in the LCC of each unrelated disease and in the AGRE LCC, we derived a highly stringent ASD-specificity protein score, allowing the prioritization of encoding genes for follow-up. Low scoring proteins were not replicated in the AGRE dataset, and were present in one or more unrelated diseases, whereas the highest scoring proteins were present in both ASD LCCs, but in none of the LCCs generated from the unrelated diseases. This analysis revealed that the majority of proteins (∼63%) were present only in the AGP network, while 31% of the proteins were present in at least one additional non-ASD network, and thus were not specific. From the 25 proteins identified in both ASD networks, the majority (56%) was not present in any ASD-unrelated network and 28% were present in one of the ASD-unrelated networks.de novo mutations in ASD , TSC2) and an increased overlap with known candidate genes ) and with genes contained in potentially relevant CNVs identified by the AGP analysis (∼26%). Enrichment in mouse phenotypes was also similar but, in addition, an enrichment in abnormal nervous system phenotype became significant, and in abnormal behavior/neurological phenotype borderline significant.Using this gene scoring system, based on gene reproducibility and specificity for ASD, we built a network with the 14 top scoring genes and their first neighbors in the LCC network . The larde novo mutation in the phosphogluconate dehydrogenase gene (PGD) has recently been reported in ASD ABL1 (Uniprot P00519), RPS6KA1 (Uniprot Q15418) and PPP1CB (Uniprot P62140) genes were identified in ASD patients from the AGP study. A query of our genes in the NEXTBIO platform, a data mining framework that integrates and correlates global public datasets with several normal and disease phenotypes, revealed correlations of six genes with ASD. For instance, deletions within the NASP (Uniprot P49321) gene were identified in ASD patients from the Simons Simplex Collection (SSC) NR4A1 (Uniprot P22736), ABI1 (Uniprot Q8IZP0), BBS4 (Uniprot Q96RK4), LMNA (Uniprot P02545) and ABL1 genes, was found in postmortem brain tissue CTSB (Uniprot P07858), BBS4, LMNA and ABL1 genes, were associated with abnormal nervous system phenotypes in animal models. The most strongly associated genes to ASD, using the AGP data, were the peroxiredoxin 1 gene (PRDX1 (Uniprot Q06830)) and cathepsin B gene (CTSB).The genes encoding the 14 top scoring proteins were considered the best candidates for harboring common variants associated with ASD risk . These gP<0.1 encoded proteins that directly interact beyond random expectation, are more rarely found isolated in the network and are connected in significantly larger LCCs than expected by chance, suggesting a functional connection. These results support recent findings from the AGP consortium, showing that stronger association of allele scores with case status was generally achieved when those scores were based on markers associated at significance thresholds higher than 0.2 P-values as high as 0.2, In this study we have conducted a network-based analysis of two ASD GWAS datasets, hypothesizing that small effect ASD risk variants hidden at the level of GWAS statistical noise can be discovered from networks of genes with related biological functions. Mapping of association data to a PPI network indeed revealed that, in both datasets, ASD-associated genes at de novo mutations have been described in whole-exome sequencing studies of nearly a thousand ASD patients de novo variants which do not explain the heritability of the disorder, but support recent observations that common and rare variants associated with ASD disturb common neuronal networks The relevance to ASD of these networks was further illustrated by their higher performance in retrieving known ASD candidates compared to top GWAS genes, and the increased similarity between the two ASD datasets, when compared to SNP or gene level overlap. Remarkably, the AGP and AGRE LCCs included 20 genes, respectively, in which de novo mutations or potentially pathogenic CNVs in ASD.As an additional filter for meaningful ASD biology, we derived an ASD candidate gene prioritization system ranking the genes encoding proteins included in the AGP LCC for ASD reproducibility and specificity. The scoring system used was very stringent, in particular since some of the control disorders are neurological  and may share susceptibility genes and pathways with autism CTSB, BBS4, LMNA and ABL1 genes have been associated with neurobiological phenotypes identified in an enrichment analysis of mouse neurobiological phenotypes from a list of 112 ASD candidate genes CTSB and ABL1 associated with cerebellum morphological and development abnormalities. The AGP genome-wide analysis identified potentially pathogenic CNVs spanning ABL1, RPS6KA1 and PPP1CB, whose relevance needs to be further established. Likewise, in the phosphogluconate dehydrogenase gene (PGD), a de novo mutation has recently been reported in a patient with ASD PRDX1, which also has an important antioxidant protective role in cells This approach generated a list of 14 top-scoring genes, present in the two ASD networks and none of the other disorders, which were considered strong candidates to harbor common variants associated with ASD risk. These genes are mostly novel candidates for ASD, and are involved in nervous system pathways or other more fundamental biological processes which have been widely associated to ASD, such as ubiquitination a priori assumptions of association thresholds. The present results have shown that autism-associated genes at higher than conventional levels of significance are functionally related, and were used to extract relevant disease biology and uncover small effect variants contributing to the disorder. The study highlighted a group of novel susceptibility genes relevant for CNS function with a high probability of bearing common variants associated with autism, which have been elusive thus far, and warranting further analysis for identification of causal variants.Thus far the use of protein networks to address common risk variants in ASD was limited to enrichment analysis of GWAS top hit genes in co-expressed or differentially expressed networks Figure S1Network properties per gene-wise P-value for each ASD dataset. For each –Log10 gene wise association P-value cutoff in the x-axis, the percentage of direct interactions (A) and isolated nodes (B) and the logarithm of the LCC size (C) were plotted for proteins encoded by disease-associated genes (red line) and for the mean of 1000 equal sized random samples of proteins (blue line). Dark grey areas represent the range between the 25th and 75th quartiles and light gray areas indicate the range between the minimum and maximum values of the random data. Empirical P-values are indicated for each gene wise association P-value comparison. Values are plotted until the –Log10 for which the percentage of direct interactions and isolated nodes reaches 0 and 100%, respectively.(TIF)Click here for additional data file.Figure S2Overlap between the two ASD datasets at SNP, gene or network level. Venn diagrams showing the overlap between the two ASD datasets (AGP and AGRE) at SNP, gene or network level.(TIF)Click here for additional data file.Table S1GWAS datasets used in the analysis and genotyping platforms.(XLSX)Click here for additional data file.Table S2AGP LCC network genes. List of the 416 genes included in the AGP LCC with information on gene-wise association P-value, specificity score for ASD and previous findings regarding implication in ASD and other neurological disorders.(XLSX)Click here for additional data file.


Multiple sclerosis is a chronic inflammatory disease of the central nervous system, characterized by an aberrant activation of the immune system and combining demyelination with neurodegeneration. Studies on experimental models of multiple sclerosis revealed immunomodulatory and immunosuppressive properties of mesenchymal stem cells. Clinical trials using mesenchymal stem cells therapy in multiple sclerosis patients showed tolerability, safety on short term, some immunomodulatory properties reducing the Th1 proinflammatory response and the inflammatory MRI parameters. The author reviews the data about experimental studies and clinical trials using mesenchymal stem cells for the treatment of multiple sclerosis.Abbreviations: MS = multiple sclerosis, RRMS = relapsing-remitting multiple sclerosis, PPMS = primary progressive multiple sclerosis, EAE = experimental autoimmune encephalomyelitis, MOG = myelin oligodendrocyte glycoprotein, PLP = proteolipid protein, MSCs = mesenchymal stem cells, CNS = central nervous system, IT = intrathecal, ALS = amyotrophic lateral sclerosis, MSC-NPs = mesenchymal stem cell-neural progenitors, EDSS = expanded disability status score. Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) leading in the end to demyelination and axonal loss. The disease is complex and multifactorial, but it seems that the main etiopathogenic event is represented by an aberrant response of the immune system cells (T and B-lymphocytes) to myelin proteins. There are three forms of the disease evolution. Approximately 80% of the patients have a relapsing-remitting form (RRMS), and two thirds of them will develop a secondary progressive form after 10-15 years from the disease onset. Approximately 20% of the patients develop a progressive form right from the onset, the primary progressive multiple sclerosis (PPMS).Experimental studies with mesenchymal stem cells1] and in vivo [2] suggested that MSCs might be effective in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, stopping the (auto) immune attack against myelin antigens and promoting nervous tissue repair by their integration into the CNS. In 2005, Zappia et al [3] demonstrated that the intravenous injection of syngeneic MSCs improves the clinical course of the experimental autoimmune encephalomyelitis induced by myelin oligodendrocyte glycoprotein (MOG) and reduces demyelination and leukocyte infiltration in the CNS. Cells have these effects only when injected at disease onset or peak; the administration during the chronic stage has no effect. A decisive finding was that T lymphocytes isolated from treated mice did not proliferate in vitro after the exposure to new MOG, an observation suggesting that MSCs induce peripheral immune tolerance. In a subsequent publication, following the intravenous of xenogeneic MSCs (human) in mice with EAE, Zhang and his team found a clinical improvement of the histological scores and some evidence of oligodendrogenesis, possibly through the secretion of neurotrophic factors [4]. In 2006, another group confirmed that the administration of MSCs protects neurons from the inflammatory attacks associated with experimental autoimmune encephalomyelitis. They also found a low degree of co-localization (less than 10%) of the labeled human MSCs with cells expressing neural markers [5]. Therefore, their findings suggested that a degree of transdifferentiation was present. A subsequent report proved that intravenous injection of MSCs in SJL mice with EAE inhibits the pathogenic humoral immune response by reducing the production of proteolipid protein (PLP)-specific antibodies [6]. Moreover, the exposure of PLP-specific encephalitogenic T cells to MSC in vitro prevents the passive transfer of EAE. Some labeled MSCs were detected in the inflamed CNS, close to the infiltrated cells, but no evidence of neural transdifferentiation was found. It is worth noting that neural [7] stem cells and, more recently, neural precursor cells derived from human embryonic stem cells [8], showed a remarkable benefit following their administration to mice with EAE through the assistance mechanism (“bystander mechanism”) that leads to the immunomodulation of autoreactivity and to neuroprotection [9]. Besides the aforementioned immunomodulatory characteristics, MSCs are endowed with many other therapeutic properties, as demonstrated by various studies.Findings about the fact that MSCs inhibit the proliferation of T cell both in vitro [10]. In addition, some research groups reported that MSCs showed a minimum potential for transdifferentiation into neurons when transplanted into the CNS of affected mice [11]. More importantly, MSCs could promote endogenous repair by recruiting local neural precursor cells, thus leading to a degree of neurogenesis and remyelination [12]. Many other features are likely to be relevant to the use of MSCs in multiple sclerosis or in other diseases of the CNS. MSCs showed a strong antioxidant effect in mice affected by EAE [13]. The neuroprotective effect may result from the release of anti-apoptotic [14] molecules and of neurotrophins [15]. These results support the idea that MSCs promote CNS repair by acting both as tolerogenic cells and as bioactive providers of trophic and anti-apoptotic factors which lead to neuroprotection [16].Despite some concerns about their transdifferentiation potential, it is accepted that MSCs can differentiate into mature mesoderm cells such as bone, cartilage and fat [Clinical trials using mesenchymal stem cells for treatment of multiple sclerosis17].The rationale for using adult stem cells in multiple sclerosis originates in the hope that they might repair the CNS by tissue integration and possibly by differentiating into neural cells. Recent evidence from preclinical studies suggested that MSCs, a sub-class of adult progenitor cells isolated from almost any mesoderm tissue and already clinically studied in various diseases, are an effective cell therapy for EAE. In EAE, intravenous injection of MSCs improves the clinical evolution of the disease and reduces demyelination, immune infiltrates and axonal loss. Surprisingly, these effects do not require grafting of MSCs in the CNS, but are based on the MSCs’ ability to inhibit the pathological response of T and B cells and on the secretion of neuroprotective and pro-oligodendrogenic molecules, facilitating tissue protection and repair. These results led to the conclusion that the administration of MSCs should focus on individuals with ongoing inflammation and before they develop irreversible disability [18]. International experts in multiple sclerosis and stem cells, together with immunologists formed the “International Stem Cells Transplantation Study Group” (IMSCTSG) in order to achieve a consensus protocol on the use of MSCs for the treatment of multiple sclerosis, with protocols for cell cultivation and patient treatment. This international consensus treatment protocol based on the efforts of Dr. Uccelli (University of Genova) and Dr. Freedman (University of Ottawa) was conducted and published in 2010 [19].The first small clinical trials in multiple sclerosis have already proven MSCs can be safely used, creating the possibility to move from the clinical use to phase II - extensive studies which address the biological effects of MSCs on markers of disease activity . A small number of multiple sclerosis patients, who obtained benefits from receiving intravenous or intrathecal infusions of MSCs, was reported. The results of a Phase I/ II study with MSCs used in neurological diseases reported the administration of autologous MSCs in 19 patients with amyotrophic lateral sclerosis (ALS) and 15 patients with multiple sclerosis [18]. MSCs were administered as a combination of intravenous and intrathecal injections, at doses of up to 60-70x 10^6 cells/ injection/ patient and the patients monitored for 6 to 28 months, mainly to determine the feasibility and safety of the procedure. None of the patients experienced any significant side effect, except for a mild meningeal irritation such as a headache and fever in those intrathecally injected with MSCs. No infection associated with the (early or late) injection was reported.Autologous MSCs derived from the bone marrow of patients with multiple sclerosis exhibit the same properties as MSCs from healthy donors, in terms of proliferation, phenotype, and differentiation in vivo and immunosuppressive ability [21] and a report on 7 patients intrathecally treated in Lebanon [22]. They did not communicate significant side effects except for those pertaining to the route of injection (different types of headaches and one case of iatrogenic meningitis) and one case of transient encephalopathy in a patient who intrathecally received an increased dose (100x10^6).Additional data available on the intrathecal use of multipotent mesenchymal stromal cells in multiple sclerosis include a study on 10 patients with progressive multiple sclerosis in Iran [23] included 10 patients with a 14.4 years average duration of disease progression, who received an intravenous dose of 1-2x10^6 cells/ b.w. The patients were clinically monitored for side effects for 4 hours after receiving the dose and also at 3 and 6 months after their treatment. The immediate adverse effects recorded were type I hypersensitivity  and were reported in 10% of the subjects after intravenous administration. A statistically significant improvement of the visual acuity and visual evoked potentials was found, as well as an increased optic nerve area following MSCs therapy. There were no identifiable effects on color vision, visual field, macular volume, retinal nerve fiber layer thickness or optic nerve magnetization transfer ratio. The authors’ conclusion was that MSCs were proven safe for use in patients with secondary progressive multiple sclerosis. The study provided evidence of structural, functional and physiological improvement with respect to several goals established for the visual function, which is indicative of the neuroprotective effect of MSCs.A phase II open-label clinical trial with MSC in secondary progressive multiple sclerosis (SPMS) which was completed and whose results were published (MSCIMS)  [24]. Investigators used the administration of autologous mesenchymal stem cell-neural progenitors (MSC-NPs), included 20 patients with progressive MS with established disability (average EDSS=6.0). MSC-NPs were administered IT in three doses of up to 10 million cells per injection, spaced three months apart. Preliminary safety outcomes in the first five study subjects indicated safety and tolerability of the treatment.A phase I open label clinical trial of intrathecal (IT) presented a preliminary result at ACTRIMS 2014 [25] conducted an open-label study including 25 patients with progressive MS (EDSS from 4.0 to 6.5) who received a single intrathecal injection of autologous MSCs and were followed-up for 12 months. Associated short-term adverse events of injection consisted of transient low-grade fever, nausea/ vomiting, weakness in the lower limbs and headache. No major delayed adverse effect was reported. 3 patients left the study for personal reasons. The mean (SD) expanded disability status scale (EDSS) score of 22 patients changed from 6.1 (0.6) to 6.3 (0.4). In the MRI evaluation, 15 patients showed no change, whereas 6 patients showed new T2 or gadolinium enhanced lesions (1 lost to follow-up). It seems that MSC therapy can improve/ stabilize the course of the disease in progressive MS in the first year after injection, with no serious adverse effects.Bonab et al (2012) [26] included 9 patients who received a dose of 1-2x10^6 MSCs/ Kg body weight intravenously infused. Immunological changes that were consistent with a lower proinflammatory T cell profile, resulting from the decrease in the proportion of IFN-c and with lesser intensity of IL-17-producing CD4+ T cells, and a reduced Th1/ Th17 ratio, were observed. From the MRI point of view, there was a significant trend to lower the mean number of GEL in the second period, suggesting a potential carryover effect of MSCs administration. There were no AE reports after the completion of the 12 months protocol.A recent randomized placebo-controlled phase II trial [A systematic review and meta-analysis of the prospective clinical trials that used intravascular delivery of MSCs  in adult populations or mixed adult and pediatric populations (SafeCell) included 36 studies with a total of 1012 participants with clinical conditions of ischemic stroke, Crohn’s disease, cardiomyopathy, myocardial infarction, graft versus host disease, and healthy volunteers. The meta-analysis of the randomized clinical trials did not detect an association between acute infusion toxicity, organ system complications, infection, death or malignancy. There was a significant association between MSCs and transient fever. However, the results from the experimental studies raised some concerns regarding the tumorigenic potential.Mesenchymal stem cells showed effects on innate and adaptive immunity in studies on animal models of multiple sclerosis. Most of the reported trials were uncontrolled open-label phase I studies and included secondary progressive MS, progressive MS, relapsing-remitting and secondary progressive MS and active but unspecific patients, a phase II trial, on secondary progressive MS which included visual and neurophysiological parameters of efficacy and one randomized controlled trial on a small number of relapsing-remitting MS patients not responding to at least one year of approved therapy. The way of administration was intrathecal or intravenous, the number of cells administered and also the number of administrations varied between trials. The patients included were with important disabilities and were followed up to 28 months. All trials reported safety and tolerability of MSCs and stabilization/ mild improvement of the disease. There are still issues to be clarified about the migration potential and homing into the central nervous system of MSCs and also about the possibility of tracking the distribution of MSCs in humans. The small number of patients included in these trials and the differences between the trial designs consist of limitations in the interpretation of data and justify the necessity for further randomized multicentric controlled trials.Source of funding: noneDisclosures: None


Self-renewal and the potential to yield various cell types are the main attributes of stem cells and shared by the so called cancer stem cells (CSCs). CSCs consist a small portion of cells within the tumor mass that are very often resistant to chemotherapy and radiation, while being responsible for cancer relapse after treatment. The role of cancer stem cells has been recognized in cancers such as leukemia and colon cancer, whereas their role in hepatocellular carcinoma (HCC) remains to be unraveled. The vast majority of HCCs (~80%) are related to Hepatitis virus B (HBV) and C (HCV) persistent infection implicating parallel pathways of intrinsic cell transformation and chronic infection-inflammation.CSCs reside among other more differentiated cancer cells, are resistant to apoptosis and their self-renewal is based on transcription factors similar to those of adult and embryonic stem cells such as Oct4, Nanog and Klf-4. Although their origin and relationship with the tumor initiating cell is still debatable it is rather established that they sustain tumor growth, produce differentiated progenies that organize a hierarchical cell system and induce eventually tumor's metastatic potential. Recently, it has been suggested that persistent oncogenic viruses such as HBV and HCV trigger the formation of CSCs that are possibly responsible for the high rates of recurrence and mortality that characterize HCC.Both HBV and HCV although very different in their genome organization and life cycle they result often in persistent infection that leads eventually in 1–5% of infected individuals to HCC through liver fibrosis and cirrhosis. HBV replicates its DNA in cell nucleus and remains episomally in the form of minichromosome. HBV DNA fragments are often integrated in host genome. HCV on the other hand is a positive strand RNA virus with cytoplasmic replication organized in virus induced vesicles. HBx protein of HBV and HCV core, NS3, and NS5A proteins are modulators of cellular pathways that lead to cell transformation. HBV and HCV chronic infection induce accumulative liver damage followed by an increasing liver regeneration that is favored by both viruses. Indeed, hepatic progenitor cells increase their numbers in the liver of HCV patients as the disease advances to cirrhosis  doublecortin and CaM kinase-like-1 (DCAMKL-1) and CD133 in conjunction with liver progenitor cell makers such as α-fetoprotein (AFP). Remarkably, curing the cells by using Interferon 2a reversed the phenomenon, while xenograft-derived tumors of these cells in mice encompassed cells with liver progenitor and CSC traits  or stemness. Such condition, which is often present in inflamed and malignant areas of the liver, is hypoxia. Accumulated evidence from various cancer types strongly support the hypothesis that hypoxia sustains the self-renewal characteristics of a portion of cancer cells in hypoxic niches mainly due to the upregulation of Oct4, NANOG, SOX2, Klf4, and c-myc (Mathieu et al., A less evaluated parameter in HCV and HBV HCC are the extrinsic signals of CSC sustainment. Cytokines such as IL-8 are secreted during HBV infection, while they fluctuate during disease exacerbation periods (Dunn et al., Thus, it is becoming increasingly supported that HBV and HCV infection promotes EMT and stem cell-like characteristics in the infected liver cells that enable them to adapt and proliferate in the hypoxic inflammatory environment of the cirrhotic liver paving the ground for the occurrence of HCC Figure . These cThe 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.


Infants who are breastfed are at an immunological advantage when compared with formula fed infants, evidenced by decreased incidence of infections and diminished propensity for long term conditions, including chronic wheeze and/or asthma. Exclusive breastfeeding reduces the duration of hospital admission, risk of respiratory failure and requirement for supplemental oxygen in infants hospitalised with bronchiolitis suggesting a potentially protective mechanism. This review examines the evidence and potential pathways for protection by immunomodulatory factors in human milk against the most common viral cause of bronchiolitis, respiratory syncytial virus (RSV), and subsequent recurrent wheeze in infants. Further investigations into the interplay between respiratory virus infections such as RSV and how they affect, and are affected by, human milk immunomodulators is necessary if we are to gain a true understanding of how breastfeeding protects many infants but not all against infections, and how this relates to long-term protection against conditions such as chronic wheezing illness or asthma. The development of the infant immune system is influenced by primary exposure to antigens that will promote either antibody dominant humoral immune responses (T helper lymphocyte (Th)2 mediated) or cytokine-cell mediated responses Th1 mediated)  mediated,4. The sInfants who are breastfed are at an immunological advantage when compared with formula fed infants, as evidenced by both decreased incidence of infections during infancy but also diminished propensity for a number of long term conditions, including chronic wheeze and/or asthma ,8,9,10. Potential immunomodulators which continue to be identified in human milk include most established mediators, such as cytokines and maternally derived leukocytes, but may also include epigenetic factors. These immunomodulators may affect development through both direct and indirect influences on the naive cells of the immature infant immune system during the early, formative stages of immune development .ex utero.Th2-type cytokines are predominant in favourable intrauterine environments of human infants . StudiesInfants born to atopic mothers are at higher risk of developing hypersensitivity than those born to an atopic father . This maLactobacillus rhamnosus (Lactobacillus GG) to mothers in the final weeks of pregnancy and continuing through 6 months of breastfeeding can halve the incidence of atopy during the first 2 years of an infant’s life [Children exposed to early infections, other than those of the lower respiratory tract, through contact with older siblings or attendance at day care or nursery facilities, prior to 6 months of age, are less likely to develop asthma than their only child, home care peers ,21,22,23t’s life , while dt’s life ,26,27,28t’s life . HoweverA link between hospitalisation with RSV bronchiolitis and the subsequent development of chronic wheezing illness has been well established ,30,31,32The mechanism by which RSV predisposes to chronic wheeze or asthma is unclear. One hypothesis centres around the extensive damage caused to the respiratory epithelium by the virus via upregulation of chemokines such as IL-8 and regulated on activation, normal T cell expressed and secreted (RANTES), which results in an influx of polymorphonuclear granulocytes, particularly neutrophils and eosinophils, into the airways ,41,42,43in vitro [Activation of the immune system by infections such as RSV and autoimmune diseases such as atopy, significantly alter the cytokine profiles produced by immunoactive cells. These changes affect cells not only at sites of activation, such as the respiratory mucosal surfaces, but also systemically in peripheral blood. The peripheral blood mononuclear cells of infants hospitalised with RSV-mediated respiratory tract infections demonstrate a significantly elevated IL-4 to IFN-γ ratio upon mitogen stimulation in vitro .+ T lymphocytes (cytotoxic/suppressor) and CD8+/CD25+ (IL2 receptor) activated T lymphocyte populations [+ (helper/inducer) T lymphocytes as being the primary effectors responsible for viral clearance rather than the CD8+ cytotoxic subset [RSV infection also alters ratios of lymphocyte subsets through an increase in B lymphocyte numbers and changes in CD8ulations ,49,50,51c subset . Murine c subset ,54.+ T lymphocyte or CD4+ T lymphocyte activity, respectively, both resulting in increased viral clearance and reduced severity of illness in RSV-infected mice [+ T lymphocytes as the major effector for viral clearance. This indicates a differing immunological mechanism working under the influence of Th1 versus Th2 cytokines in RSV infection.RSV infection upregulates both IL-10 mRNA and protein expression by lung alveolar macrophages, as well as increasing the ratio of IL-4/IFN-γ production by PBMC ,55,56. Tted mice . IntramuIt is therefore feasible that maternal RSV infection will exert similar effects on the cells and cytokines found in human milk, manifesting in changes to the Th1/Th2 balance and ultimately in differing responses in the infant.Breastfeeding confers protection against both incidence and severity of RSV disease, particularly in those born prematurely, as well as the subsequent development of recurrent wheezing illness ,63,64,65Breastfed infants with RSV infection have markedly higher serum IFN-α levels than RSV infected non-breastfed infants during the course of the illness and continuing throughout the first 2–4 weeks post-infection . This ma+ and CD25+), and memory (CD45RO+) T cells than peripheral blood [ex vivo [in vitro, indicating a comparable response by human milk-derived leukocytes to that from circulatory leukocytes [The lymphocyte population of human milk consisting largely of T cells is made up of significantly more activated  prevalent in both the gastrointestinal and respiratory mucosa provide an intrinsic link between innate and adaptive immunity which is substantively regulated by the microenvironment at the time of antigen recognition. Resultant differences cytokine and in cell surface molecule expression, including toll-like receptors (TLRs), determine DC effects on the development of T-cell sub-populations (Th1/Th2/Th17 and regulatory T cells) and thereby antigen-specific activity by T cells. While mature DC have not been identified in human milk, the potential for breast milk macrophages to commence this transition has been demonstrated ex vivo . These bex vivo . As RSV ex vivo .Corynebacterium and Dolosigranulum a newly recognized member of the family of lactic acid bacterium, generally considered to be contributors to a healthy microbiota. Conversely formula fed infants had decreased Staphylococcus, and Prevotella and Veillonella, anaerobic bacteria, recently associated with an increased risk of otitis media [Dolosigranulum in the nasopharynx and parent reported symptoms of wheezing and respiratory infections was observed. The reported effect was absent by 6 months of age suggesting a limited window of direct benefit. However, this study provides an interesting beginning to this novel area of investigation and does not preclude the possibility for early immunomodulation by favorable microbiota extending beyond this window of difference.An association between the microbiota of human milk and of the infant gut has been established ,93. Moreis media . While tin vitro however a single clinical study describing differentiation of miRNA response not just to RSV infection but also relating to disease severity marks this as a promising novel avenue for the future [Finally, miRNA expression has been established in human milk, including identification of miRNA’s unique to milk and large quantities of immune-related miRNAs particularly in the first six months postpartum ,96,97. Ae future .A link between infant and mother through milk feeding is well accepted. However, evidence supporting the potential for that link to be bi-directional has only recently emerged. Maternal exposure to RSV during their infant’s disease course significantly increases the number of cells passed from the mother to her infant in milk . In addiThis research provides evidence for a direct link between the cells found in human milk and illness in the recipient infant, regardless of the presence of clinical symptoms in the breastfeeding mother. The marked increase in cells, which are skewed to a Th2 cytokine response to RSV, may affect the outcomes of RSV disease and the development of asthma in the recipient infant. Determining the degree and specificity of activation of human milk cells that traverse the mammary gland during RSV infection will help determine the potential for these cells in activating and modulating the developing immune system of the recipient infant.Further investigations into the mechanisms by which respiratory viruses such as RSV affect human milk immunomodulators is necessary if we are to gain a true understanding of how breastfeeding protects many infants against infections, as well as associated long-term immune-mediated conditions such as chronic wheezing illness or asthma. This future research must also examine why, for some infants this protection is incomplete. In this light, the composition of human milk must be profiled further. This is a continuing process with new cytokines being identified in samples of human milk almost as soon as they are identified in blood. Similarly, the cells of human milk are currently being identified and characterised by many researchers and this process must continue, including broadening to encompass the latest understanding of leukocyte cell subsets such as the T cell Treg and Th17 and monocyte/macrophage M1 and M2 if we are to understand the potential influence of these potent immunomodulators.Identifying the potential of these human milk cells, as well as the complete cytokine profile of milks in response to immuno-active conditions, will also provide the first steps toward furthering research into modification of these factors. While cellular activation, as well as the production of many cytokines and prostaglandins, can be regulated by pharmaceuticals, in the context of infant feeding a preferable option would always be less synthetic means such as nutriceuticals including the pro-biotics ,99,100. Overall, we have merely scratched the surface in the process of gaining an understanding of what is in milk, how it gets there and what it does in the context of respiratory viral illnesses so often shared by mother and child. Extensive further research is required if we are ever to truly appreciate the benefits available to all infants via this paradoxical fluid.


In early stages of life, this tolerogenic state of the innate and adaptive immune system and the lack of immunological memory render the host more susceptible to infectious pathogens like HIV. HIV pathogenesis is different in children, compared to adults, with more rapid disease progression and a substantial lack of control of viremia compared to adults. Plasma viral load remains high during infancy and only declines gradually over several years in line with immune maturation, even in rare cases where children maintain normal CD4 T-lymphocyte counts for several years without antiretroviral therapy (ART). These pediatric slow progressors also typically show low levels of immune activation despite persistently high viremia, resembling the phenotype of natural hosts of SIV infection. The lack of immunological memory places the fetus and the newborn at higher risk of infections; however, it may also provide an opportunity for unique interventions. Frequencies of central memory CD4+ T-lymphocytes, one of the main cellular reservoirs of HIV, are very low in the newborn child, so immediate ART could prevent the establishment of persistent viral reservoirs and result in “functional cure.” However, as recently demonstrated in the case report of the “Mississippi child” who experienced viral rebound after more than 2 years off ART, additional immunomodulatory strategies might be required for sustained viral suppression after ART cessation. In this review, we discuss the interactions between HIV and the developing immune system in children and the potential implications for therapeutic and prophylactic interventions.The developing immune system is adapted to the exposure to a plethora of pathogenic and non-pathogenic antigens encountered  In the dynamic developmental period from fetal life to adolescence, the human organism is more susceptible and vulnerable to chronic viral infections . HIV patHowever, the recent report of a child who apparently achieved “functional cure” for more than 2 years following early therapeutic intervention  has raisOf the estimated 35 million people living with HIV today, 3.3 million are children (UNAIDS 2013 report). Administration of combination antiretroviral therapy (ART) can reduce risk of transmission to approximately 1–2%  and WHO in utero (5–10%), intrapartum (15%), or post-partum via breastfeeding (15%)  . Interesng (15%) . During  utero 5–%, intrapng (15%) , since tng (15%) . Indeed,ng (15%) . In humang (15%) . However% . Interin utero, intrapartum and postnatal infection found median survival times from infection of 208, 380, and >500 days, respectively  through pattern recognition receptors (PRRs) that have evolved as substrate-specific receptors that can discriminate between self- and non-self and constantly sample the intra- and extracellular milieu [reviewed in Ref. ]. PRRs iThe importance of these innate immune mechanisms in viral infections has been demonstrated in children with gene mutations in the TLR3 signaling pathway, who are prone to severe herpes simplex encephalitis . Certainin utero and after birth when the newborn is exposed to an abundance of newly encountered antigens; and second, to ensure protection against extracellular bacterial and fungal pathogens, which can be rapidly fatal soon after birth , dendritic cells, and innate lymphoid cells , 164 disThe immaturity of the gut in early life, characterized by increased epithelial permeability  and the In vertically HIV-infected infants, levels of microbial translocation are elevated compared to uninfected controls and persist at a higher level after initiation of ART . PersistHIV-specific antibodies have been in the focus of intense research for several decades in the quest for a prophylactic HIV vaccine. Recently, hope has been fueled by the results of the RV144 vaccine trial, which showed modest vaccine efficacy  and idenThese broadly neutralizing antibodies (bNABs) have been shown to occur naturally in a minority (20%) of HIV-infected subjects over the course of several years post-infection –182 duriin utero HIV-infected child who appeared to have been “functionally cured” after early initiation of ART was received with much excitement. The child initiated ART within 30 h of life and stayed on ART until 18 months of age when ART was discontinued by the mother. In subsequent visits to the clinic half a year later and in the successive follow-up period, viral load levels remained undetectable in the absence of ART and no signs of ongoing viral replication or HIV-specific T-cell or antibody responses were detected (The case report of an detected , hence tThe main obstacle to HIV eradication is a pool of long-lasting, treatment-resistant viral reservoirs that only decay slowly on ART and are a source for viral rebound after treatment cessation . DThe frequency of latently infected CD4 T cells in children strongly depends on early initiation of ART and the time to achieve virologic suppression . A crossApart form timing of ART initiation, the size and constitution of the viral reservoir is affected by host immunological factors including persistent immune activation that fuels replenishment of the reservoir during ART [reviewed in Ref. ] and effin utero and after birth. The initial ontogeny of the immune system is characterized by a more tolerogenic state of innate and adaptive immunity that prevents potentially harmful autoimmunity and inflammatory responses, and is geared toward preferential protection against extracellular pathogens. As a consequence, vertically acquired HIV infection is poorly controlled and marked by rapid progression to AIDS and death without effective interventions. Chronic infection results in systemic immune activation that drives immunopathology with functional immune exhaustion, increased susceptibility to co-infections, and inflammatory comorbidities. Coverage rates of PMTCT, early infant diagnosis, and pediatric ART need to be improved worldwide to reduce the global disease burden among children. For those children who become infected despite PMTCT, novel therapeutic strategies need to be developed targeting persistent immune dysregulation and the viral reservoir to potentially achieve drug-free remission.In summary, immunity to HIV in early life differs from that in adulthood not only in quantity but also in quality. This results from the adaptations of the immune system to the abundance of antigenic stimulation and pathogen exposure 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.


Salinity stress is a major constrain in the global rice production and hence serious efforts are being undertaken towards deciphering its remedial strategies. The comparative analysis of differential response of salt sensitive and salt tolerant lines is a judicious approach to obtain essential clues towards understanding the acquisition of salinity tolerance in rice plants. However, adaptation to salt stress is a fairly complex process and operates through different mechanisms. Among various mechanisms involved, the reactive oxygen species mediated salinity tolerance is believed to be critical as it evokes cascade of responses related to stress tolerance. In this background, the present paper for the first time evaluates the ROS generating and the scavenging system in tandem in both salt sensitive and salt tolerant cultivars of rice for getting better insight into salinity stress adaptation.2O2) and lower level of superoxide ions (O2-) in the salt tolerant as compared to salt sensitive cultivars. Specific activity of ROS generating enzyme, NADPH oxidase was also found to be more in the tolerant cultivars. Further, activities of various enzymes involved in enzymatic and non enzymatic antioxidant defence system were mostly higher in tolerant cultivars. The transcript level analysis of antioxidant enzymes were in alignment with the enzymatic activity. Other stress markers like proline were observed to be higher in tolerant varieties whereas, the level of malondialdehyde (MDA) equivalents and chlorophyll content were estimated to be more in sensitive.Comparative analysis of ROS indicates the higher level of hydrogen peroxide (H2O2 which is considered as a key determinant for conferring salt stress tolerance to rice might have originated through an alternative route of photocatalytic activity of chlorophyll.The present study showed significant differences in the level of ROS production and antioxidant enzymes activities among sensitive and tolerant cultivars, suggesting their possible role in providing natural salt tolerance to selected cultivars of rice. Our study demonstrates that the cellular machinery for ROS production and scavenging system works in an interdependent manner to offer better salt stress adaptation in rice. The present work further highlights that the elevated level of H Rice is an important cereal crop that has the potential to provide food security to the increasing world population. Out of the total global agricultural land, 150 million hectares is estimated to be under rice cultivation that leads to an annual production averaging to 500 million metric tons , 3. Amon2-), hydroxyl radical (OH-) and hydrogen peroxide (H2O2) are produced in a low concentration as a result of normal cellular metabolism in plants and plays important role in growth, development and in adaptation to stress -3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzenesulfonic acid hydrateAPX, ascorbate peroxidase; CAT, catalase; DAB, 3, 3-diaminobenzidine; DHAR, dehydroascorbate reductase; GPX, guaicol peroxidase; GR, glutathione reductase; H


Gap junctions (GJs) are conglomerates of intercellular channels that connect the cytoplasm of two or more cells, and facilitate the transfer of ions and small molecules, including second messengers, resulting in metabolic and electrical coordination. In general, loss of gap junctional communication (GJC) has been associated with cellular damage and inflammation resulting in compromise of physiological functions. Recently, it has become evident that GJ channels also play a critical role in the pathogenesis of infectious diseases and associated inflammation. Several pathogens use the transfer of intracellular signals through GJ channels to spread infection and toxic signals that amplify inflammation to neighboring cells. Thus, identification of the mechanisms by which several infectious agents alter GJC could result in new potential therapeutic approaches to reduce inflammation and their pathogenesis. Herpes simplex virus-2 (HSV-2) reduces expression of connexins and GJC by direct tyrosine phosphorylation of Cx43 , and attempts have been made to utilize the multipotential nature of these cells to repair and repopulate areas of CNS damage , amyotropic lateral sclerosis, and Alzheimer’s disease, but the contribution of infectious agents upon demyelination has not been examined.Oligodendrocytes are responsible for myelinating neurons in the CNS, and astrocytes provide metabolic support of this process. Homotypic Cx47 and Cx32 GJs couple oligodendrocytes to themselves, and heteromeric Cx47/Cx43 GJs couple oligodendrocytes to astrocytes Figure . Double In vivo activation of oligodendrocyte TNFα receptor induces severe myelin vacuolization and death , and induce oligodendrocyte progenitor cell (OPC) death by releasing TNFα  immunized with MOG (to induce neuroinflammation), found CD11c+ DCs abundantly infiltrated CNS parenchyma, were positive for myelin antigen after 21 days, and participated in cross-presentation with T-cells  play an essential role in generating and adaptive immune response for clearing bacterial and fungal infections through phagocytosis, antigen presentation, and release of factors that reduce inflammation. Cx43 is essential for phagocytic activity in peritoneal macrophages as seen in a mouse model implementing immunostaining and confocal microscopy of Cx43, LAMP-2, and dyna beads  and selecting for varying degrees of expression of each protein revealed that bystander cell death due to metabolic stress was in direct proportion to the number and density of GJs with less resistant neighbors  mediated signaling. CytC is a pro-apoptotic signaling molecule that mediates caspase activation by participating in apoptosome formation, and microinjection of CytC in cultured astrocytes induces 100% apoptosis. However, microinjection of CytC into HIV infected astrocytes did not induce cell death, suggesting that HIV infection protects astrocytes from apotopsis  proteins have been identified as capable of allowing the release of CytC. Bcl-2 associated X (Bax) protein is a pro-apoptotic cytosolic member of the Bcl-2 family that translocates to the OMM and is associated with CytC release and apoptosis , and donates an electron to complex IV for the final step in HUse of the retroviral beta-galactosidase at gag (BAG) vector and Herpes Simplex Virus Thymidine Kinase (HSV-tk) has expanded our knowledge of the relationship between GJIC and bystander cell death. Several studies validate the use of this retroviral vector in drug delivery intending to preserve brain tissue while treating gliomas, transfected a retroviral BAG vector into rat brain inoculated with a C6 glioma cell line  engineered to secrete IL-12, and bone marrow derived stem cells (BMSCs), have shown the ability to track migrating tumor cells (Ehtesham et al., We have outlined the involvement of Cx expression and GJC in neuronal plasticity, myelination, migration, stem cell differentiation, and discussed the devastating effects inflammation and infectious diseases can incur on these processes. Inflammation induced by disease is amplified by the opening of uHCs in microglia, and is followed by peripheral immune invasion into the CNS to resolve the damage. Adaptive immune responses following inflammation in the CNS are also dependent on Cx expression and GJC, and the mechanisms of adaptive immune response beyond antigen cross-presentation are an active field of study. In contrast, GJs are also used by some pathogens to induce bystander killing, and this phenomena has been explored to treat malignant gliomas in a novel HSV-tk/Cx43 “hunter-killer” tactic after resection. Thus, further studies to control GJC and uHCs can improve the detrimental effects of infectious diseases and associated inflammation in the CNS.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.


SOCS1 or PTPN1 [JAK2 amplifications [STAT6 gene in the exons encoding the DNA binding domain are detected in about 35% of the cases and in MedB-1 PMBL derived cell line, but their functional consequences are so far unknown [SOCS1, PTPN1, JAK2 and STAT6 overlap in some tumors, suggesting that their effects may not be strictly redundant.Primary Mediastinal B-cell Lymphoma (PMBL) is a specific subtype of B-cell lymphoma which shares some phenotypic and genotypic features with diffuse large B-cell lymphoma (DLBCL) and with classical Hodgkin's lymphoma (HL). The genomic landscape of PMBL has not been fully elucidated, but has revealed the recurrent alteration of genes involved in JAK-STAT signaling and in immune surveillance . The actor PTPN1  and to Jications . Mutatio unknown . Curiousin vitro and in vivo, and to induce a broad inhibition of STAT proteins phosphorylation [The optimal treatment strategy of PMBL remains to be precisely defined. The results of a recent trial indicate that dose intensive chemotherapy (dose adjusted EPOCH) combined with rituximab may be the best option nowadays, limiting the need of consolidation radiotherapy and the risk of associated sequelae . An alterylation . Hence, in vitro to specific inhibitors is a strong indicator that the targeted pathway is biologically relevant. Nevertheless, a lot of questions still remain: which target will be the most efficient to kill tumor cells without major side effects: JAK2 or STAT6? Why does STAT6 knock-down affect drug sensitivity in some cell lines but not in others: is it related to the presence of STAT6 or any other gene mutations? Should these targeted therapies be given alone, or in association with immuno-chemotherapy? And will these strategies prove more efficient than dose adjusted EPOCH-rituximab? Last but not least, as mentioned previously, PMBL are also characterized by the alteration of the immune surveillance system. It would be worth evaluating the relationship between JAK-STAT activation and the immune privilege of these tumors, because it could impact patients' outcome upon targeted therapy.The response of cell lines cultured


Understanding neural injury in hydrocephalus and how key physiological factors changes during the course of the disease in-vivo remain unclear. This study aims to describe brain deformation, microstructural and mechanical properties changes during obstructive hydrocephalus (HCP) development in a rat model using multimodal magnetic resonance (MR) imaging.Hydrocephalus was induced in 8 Sprague-Dawley rats by injecting a kaolin suspension into the cisterna magna. 6 sham-injected rats were used as controls. MR imaging  was performed 1 day before, then at 3, 7 and 16 days post injection. T2-weighted MR images were collected to quantify ventricular enlargement and brain deformation. MR elastography was used to measure the brain stiffness, and diffusion tensor imaging (DTI) was conducted to observe brain tissue microstructure. Histology examination was performed immediately after the last MR scan. Generalized estimating equations (GEE) were used to assess linear relationships between variables while accounting for repeated measure design.The enlargement of the ventricular system was associated with a decrease in the cortical gray matter thickness and basal ganglia cross-sectional area from day 3 , an alteration of the corpus callosum microstructure and rearrangement for the cortical gray matter microstructure , while a compression without gross microstructural alteration was evident for the basal ganglia and internal capsule , not seen in controls. Specifically, during hydrocephalus development, increase in space between the white matter tracts was observed in the corpus callosum (P<.001), while a decrease of space was observed for the internal (P<.001). For the cortical gray matter, an increase in extracellular tissue water originating from the ventricles via the discontinuous ependyma was observed and significantly associated with a decrease in stiffness (P=.001). Finally, for the basal ganglia, results suggested a progressive compression of the tissue, beginning with the periventricular region, then extending to the entire structure. The basal ganglia did not appear to be edematous, and the tissue compression was associated with an increase in stiffness (P=.001).This study outlines the temporal changes in tissue microstructure, water content and stiffness in different brain regions using DTI and MR elastography and how these changes are associated with ventricular enlargement. It shows that the effect of ventricular enlargement is not limited to periventricular white matter, severity of microstructural changes vary with brain region and there is regional and temporal variation in brain tissue stiffness during hydrocephalus development.