The Juvenile Diabetes Research Foundation (JDRF) Network for Pancreatic Organ Donors with Diabetes (JDRF nPOD) was established to obtain human pancreata and other tissues from organ donors with type 1 diabetes (T1D) in support of research focused on disease pathogenesis. Since 2007, nPOD has recovered tissues from over 100 T1D donors and distributed specimens to approximately 130 projects led by investigators worldwide. More recently, nPOD established a programmatic expansion that further links the transplantation world to nPOD, nPOD-Transplantation; this effort is pioneering novel approaches to extend the study of islet autoimmunity to the transplanted pancreas and to consent patients for postmortem organ donation directed towards diabetes research. Finally, nPOD actively fosters and coordinates collaborative research among nPOD investigators, with the formation of working groups and the application of team science approaches. Exciting findings are emerging from the collective work of nPOD investigators, which covers multiple aspects of islet autoimmunity and beta cell biology. The disease is often diagnosed in childhood or adolescence, but is also common in adult individuals . These oRecover organs from deceased donors with T1D  and establish a repository of pancreas and other relevant tissues Distribute donor specimens to approved investigators, worldwide, to support comprehensive and diversified studies of human T1DPromote and coordinate collaboration and team science approaches, by establishing and managing project interactions and focused working groups, to achieve the most comprehensive understanding of human T1DThe JDRF nPOD has three main strategic goals:This article provides an overview of the JDRF nPOD, both in terms of its operational models and the discoveries of nPOD investigators that have been recently contributed to the scientific literature.http://www.jdrfnpod.org/for-investigators/standard-operating-procedures/) [The nPOD operational model and standard operating procedures (SOP) have been recently described in detail , 5 and aedures/) . TissuesnPOD actively seeks organ donors with T1D, with particular emphasis on those with shorter disease duration that are more likely to still have detectable islet autoimmunity and residual insulin-positive beta cells. Recent onset T1D may sometimes be diagnosed when a patient has developed diabetic ketoacidosis (DKA) and has been admitted to an emergency room; nPOD established a partnership with the Pediatric and Critical Care sections of the American College of Emergency Physicians (ACEP), to engage pediatric physicians in the referral of thankfully rare but unfortunate deaths resulting from complications of DKA , 9.To identify donors who might have had asymptomatic prediabetes, characterized by islet immune infiltrates (insulitis) in the absence of diabetes symptoms, nPOD has enabled several OPOs to screen non-diabetic donors for autoantibodies associated with T1D risk. nPOD provides the OPO clinical laboratories with training, protocols, and reagents so that their staff can rapidly (3 h) screen donors for autoantibodies to the GAD65, IA-2, and ZnT8 autoantigens ).http://www.jdrfnpod.org/for-investigators/donor-groups/). nPOD also recovers donors with type 2 diabetes (T2D), as a comparison to T1D donors and to study pathways of beta cell physiology and pathology that can also be relevant to T1D. Finally, nPOD recovers organs from donors affected with gestational diabetes and cystic fibrosis; non-diabetic donors are also accepted as controls.The nPOD repository currently includes specimens from 100 donors with T1D, including donors with recent onset diagnosis, and from 22 non-diabetic donors with islet autoantibodies. At least 19 donors have been identified with insulitis in their pancreas, including donors of pediatric age. An updated list of donors is maintained on the nPOD website , depending on the investigator’s affiliation. The nPOD TPC will typically approve projects that have scientific merit and are feasible, and often will provide critical guidance and constructive feedback, both in terms of scientific design and strategic considerations that take into account sample availability and relevance to the study. Investigators are also prompted to embrace nPOD policies about data sharing and collaboration (described below). Indeed, once a project is approved, investigators are encouraged to take an active role in nPOD research activities, beyond the conduct of their own study. In fact, they are made aware of collaborative opportunities within the network, either with specific investigators or with working groups. The nPOD staff promotes interactions among investigators by organizing periodic web-based meetings (webinars) to review data and discuss scientific projects or provide training on the use of the online pathology system. In addition, nPOD holds an annual investigators’ meeting where data, most often unpublished, are presented. Indeed, nPOD is implementing “real-time” data sharing to best take advantage of the coordinated analyses of donor specimens by multiple investigators. Thus, investigators are given the opportunity to share results prior to publication, among members of the nPOD network, and this facilitates changes in the experimental strategy, reconciliation of discordant findings, as well as case selection during the course of a project. Data sharing is made possible by the creation of nication .Moreover, nPOD has developed collaborative working groups focused on major research areas where questions are many and complex and will likely require a team science approach for significant progress to be made. Thus, interested investigators begin webinar discussions of a major topic and collectively define critical questions to address in the context of a collaborative experimental strategy where synergies can be exploited. For example, nPOD has established working groups on viral etiology (nPOD-Virus), autoreactive T cells (nPOD-T), and extracellular matrix components (nPOD-Matrix), and a group on beta cells (nPOD-Beta Cell) is developing. Since inception, nPOD has provided tissue specimens in support of about 130 projects led by investigators around the world. nPOD-supported projects have a broad scope and cover critical research areas relevant to T1D, including islet autoimmunity, environmental factors, beta cell physiology and dysfunction, as well as pancreas development and remodeling, with emphasis on beta cell regeneration, trans-differentiation, and dedifferentiation. nPOD samples are analyzed with diverse and advanced methodological approaches for immunohistochemistry, immunofluorescence, electron microscopy, proteomics genotyping, RNA sequencing, gene expression, multi-parameter flow cytometry, and more.On post-transplant follow-up, reappearance of autoantibodies, months to more often years prior to diabetes recurrence, correlating with increased risk of diabetes recurrence , 16, 17At presentation of diabetes recurrence, severe, insulin-requiring hyperglycemia, in the absence of rejection and stable exocrine pancreas (urine amylase) and kidney (serum creatinine) graft function, yet in the presence of selective loss of C-peptide secretionIn the pancreas transplant biopsy, the presence of insulitis and/or beta cell loss, at the same time when islet cell autoantibodies and autoreactive T cells are present in the circulation and in the pancreas transplant-associated lymph nodes The presence of autoreactive T cells in the circulation that correlated with disease activity and progression; in fact, in patients that received additional immunosuppression in the attempt to salvage the residual B cell mass demonstrated at biopsy, autoreactive T cells were no longer detected after treatment but reappeared on later follow-up. Their return was followed by further and complete loss of C-peptide . MoreoveThe demonstration that antigen-specific autoreactive T cells isolated from these patients could mediate beta cell destruction when transplanted with human islets in immunodeficient mice. Moreover, the T cell receptor sequences (V-beta and CDR3) in samples collected over time suggested a memory phenotype , 18 and Studies we conducted at the University of Miami in T1D recipients of simultaneous pancreas-kidney (SPK) transplants have revealed that approximately 5–6 % of such patients will, on long-term follow-up, develop recurrence of T1D in the pancreas allograft , 11. WhiImportantly, the biopsies of the transplanted pancreas in our cases with diabetes recurrence revealed insulitis and beta cell loss that did not appear dissimilar in severity and pathological features from nPOD donors with spontaneous disease in whom insulitis is present in their native pancreas. Figure Studies of biopsies from pancreas transplant recipients with recurrent disease can also be relevant to study beta cell regeneration and other mechanisms of pancreas remodeling. Pancreas transplant biopsies from several of our patients with recurrent autoimmunity revealed the presence of ductal cells expressing insulin . These cIn 2012, based on the above considerations, nPOD initiated a programmatic expansion, called nPOD-Transplantation, in which patients with T1D who are recipients of pancreas transplants consent for directed, postmortem donation to the nPOD program. This new program is pioneering an approach that allows T1D recipients of pancreas allografts to consent for postmortem donation of both their transplanted and native pancreata to nPOD, so that their organs can support diabetes research  would have insulitis in the pancreas , 48, 49.Examination of nPOD specimens also shows that human insulitis is rarely as severe as reported in the NOD mouse, an experimental model that presents striking similarity with the human disease , even inStudies on nPOD cases with insulitis reveal that islet autoantigen-specific CD8 T cells are present in the insulitis lesion, using MHC tetramers to identify such cells in the pancreas of nPOD donors. These findings place those autoreactive T cells at the crime scene and as such provide as much causal evidence as is possible through the study of pathology specimens. Thus, those autoreactive T cells and antigen specificities can be considered validated therapeutic targets ••. FurthAire transcription factor, also known as the autoimmune regulator [Aire-expressing cells [Another critical area of study deals with how the immune system loses tolerance to islet cell autoantigens. The discovery that insulin and other self-molecules are expressed in the thymus in early life linked this mechanism to tolerance, and genetically determined influences of levels of autoantigen expression were linked to T1D risk and enhanced probability of generating autoreactive T cells –59. In tegulator . Followiegulator . The anaegulator ; such a egulator . These segulator  and impang cells  that areng cells •.nPOD studies show that beta cell loss occurs with distinct patterns, indicating that the disease pathogenesis is likely heterogeneous and that more than one pathway may explain beta cell death . HoweverNovel molecules have now been associated with disease, including selected chemokines . Key medThe nPOD-Virus group is conducting extensive studies seeking to address key questions about the role of viruses in T1D etiology. Ongoing analysis of the nPOD samples is providing additional evidence for an association of enteroviruses with T1D •. ImportnPOD investigations are exploring the extent to which beta cell replication might be possible in the human pancreas. Beta cell replication appears more sustained in early life. While replication is possible in adult patients, it is rare . HoweverThe analysis of the pancreas body weight revealed that T1D pancreata have reduced weight compared to those of non-diabetic donors. Importantly, the pancreas weight was also reduced in donors with autoantibodies . These fDuring the last few years, many papers have reported novel findings based on the study of nPOD samples •, 47•. MIn closing, the JDRF nPOD is actively promoting the study of T1D focusing research on the analysis of pancreas and other relevant tissues from patients. Through its interactions with OPO and transplant programs, nPOD and nPOD-Transplantation provide investigators with the rare opportunity to study specimens with ongoing disease and compare findings at different time points in the natural history of the disease and after transplantation, when the immunosuppression cover may provide additional insight into the immune responses and possibly regeneration. nPOD is pioneering an expanded team science approach, involving the coordination of collaborative efforts and real-time data sharing in the joint study of a cohort of T1D donors. Studies supported by nPOD are shedding new light into the complexity and heterogeneity of the human disease and is challenging the concept that beta cell destruction is completed and autoimmunity almost exhausted within a few months from diagnosis. nPOD can also help in validating therapeutic targets and in this function contribute to defining strategic direction and priority for clinical trials to interdict T1D.


AbstractHypothesis: Even at the beginning of the new millennium, secondary peritonitis presents a common life-threatening condition associated with high mortality and morbidity. Objective: This article comments on epidemiology, diagnosis and general principles of surgical management in patients with secondary peritonitis.  Methods and Results: The demographic data, clinical findings and surgical outcome of 204 patients who had a confirmed generalized secondary peritonitis were analyzed retrospectively. Our approach was laparotomy, surgical control of contamination, antibiotic therapy and modern intensive care support. Acid peptic disease was the most common cause of perforation peritonitis 60 , following by the perforated appendicitis 45 . The faecal peritonitis and colon perforation were found in 42 patients . The morbidity rate was 50%; 41  patients had more than one complication. The morbidity rate was significantly the highest in patients with colon perforation  . The overall mortality rate in our study was 8,82%. The mortality rate was significantly the highest among the patients with mesenteric ischemia in 4 patients , followed by colon perforation in 10 cases , and 4 deaths due to gastro-duodenal perforation . Discussion: This study has confirmed that the clinical presentation and outcome of the secondary peritonitis depend on duration of abdominal infection, the site of perforation and the general condition of the patient. Rapid surgical source control, modern intensive care and sepsis therapy may offer the chance of decreased morbidity and mortality of the intra-abdominal infections. Abbreviations  Intensive Care Unit (ICU), Perforated Diverticular Disease (PDD), Inflammatory Bowel Disease (IBD), Multiple Organ Dysfunction Syndrome (MODS), Acute Respiratory Distress Syndrome (ARDS), Nonsteroidal Anti-inflammatory Drugs (NSAID), Peptic Ulcer Disease(PUD), Ultrasonography (US), Computer Tomography (CT), Colorectal Cancer (CRC), Postoperative Adhesive Disease (PAD), Acute Mesenteric Ischemia (AMI). With a mortality of up to 20% peritonitis is a dominant cause of death due to surgical infections [1]. Generalized secondary peritonitis is one of the most common surgical emergencies. Despite the great progress in intensive care support, antimicrobial therapy and surgical techniques, the management of peritonitis is still highly complex and represents a challenge for clinicians [3]. The causes, frequency and consequences of acute abdominal surgical emergencies due to peritonitis are different. Yet most of these are elderly patients, 60 years of age and above, with severe concomitant disorders and poor conditions. In recent decades, life expectancy has increased significantly and it may consequently be expected that demand for surgical care of the elderly is rising and will rise in the future [1]. Surgical source control is the most important determinant for survival and has to be placed on top of the therapeutic priority list. Nevertheless the therapy of sepsis requires state of the art intensive care. The mortality rate increases with the length of interval between the time of hollow organ perforation and time of surgery.[1] A prompt diagnosis and urgent surgery is life-saving for all patients with generalized secondary peritonitis . Intra-abdominal sepsis, although it affects all age groups, takes a greater toll on the elderly population than it does on younger populations [4]. The signs and symptoms are usually typical for acute abdomen and it is possible to make a rapid clinical diagnosis of peritonitis in most patients. Unfortunately, in majority of cases the presentation to the hospital is late, with well established generalized peritonitis, purulent contamination and varying degree of septicemia. A loss of physiologic reserve, together with concomitant systemic illness, results in worse outcomes for generalized peritonitis, especially in elderly population, those with imunosupresion and serious comorbid condition [5]. Pathophysiology of generalized peritonitis implies the complex processes in each organ system in intra-abdominal sepsis that deplete physiologic reserves and they inhibit the ability to localize, combat, and eradicate infections [5]. Secondary peritonitis is common surgical emergency which presents a life-threatening condition associated with a high mortality and morbidity. Rapid source control combined with antibiotic therapy, modern intensive care and sepsis therapy are decisive for the outcome . Additional surgical procedures for complicated PUD include vagotomy and pyloroplasty, vagotomy and antrectomy with gastroduodenal reconstruction, or a highly selective vagotomy [8].  We found that patients with perforated gastroduodenal ulcers usually had a short history of pain starting in upper abdomen along with generalized abdominal tenderness. Thirty-nine (65%) of those patents had positive history of nonsteroidal anti-inflammatory drugs (NSAID) consumption. On admission 40 (66%) patients had evidence of pneumoperitoneum on abdominal X-Ray done in erect posture. These patients come to the hospital fast and dramatic symptoms and signs allow the rapid diagnosis. It is known that with the success of medical therapy, surgery has a very limited role in the management of peptic ulcer disease (PUD) today. The one of indications for urgent surgery in PUD is perforation. Many authorities recommend simple over sewing of the ulcer, with treatment of the underlying H pylori infection [10]. When the diagnosis is unclear, ultrasonography (US) and in particular a computer tomography (CT) are useful [10]. The mortality rate for appendicitis in the elderly population is 2%–14%, and morbidity is 40% [5].  Acute appendicitis and appendicular perforations, addition to the typical presentation, unfortunately, may have an atypical clinical symptoms and sings. In our study among patients with appendicle perforations, 33% patients had characteristic pain, localized tenderness in right iliac fossa along with vomiting and fever. Delays in diagnosis are more common in the elderly population, in patients who are mentally confused, obese, or immunosuppressed and probably account for the higher incidences of generalized peritonitis and death . In symptomatic cases, 0.5% will require surgery due perforated sigmoid diverticular disease (PDD) that result in generalized peritonitis [13]. Treatment guidelines for the PDD recommend Hartmann's procedure or anastomosis with protective colostomy for Hinchey stage III diverticulitis and Hartmann's procedure only for Hinchey stage IV diverticulitis [13]. Richter and al showed that one-stage sigmoid resection with primary anastomosis can be safe in nearly 90% of all patients with Hinchey III/IV stage [14]. However, in hemodynamic unstable patients, immunosuppression, feculent peritonitis with open communication with bowel lumen, and questionable viability of the bowel wall, single-stage operation is contraindication. The two-stage procedure should be used in these situations [13].  Colonic diverticular disease is common in Western countries, almost routinely found in elderly persons and the majority of patients (80-85%) will remain asymptomatic throughout their life [17]. Perforated colon perforation have a poor outcome, due to patients nutritional status, peritoneal spread of the cancer during perforation, and the emergent interventions in less prepared patients [17]. The common clinical presentation of colon cancer preceding perforation are abdominal pain and rectorrhagia. If the lesion is localized at or proximal to the splenic flexure, recommendation is one-stage resection and right or extended right colectomy with ileocolic anastomosis [17]. In the presence of lesions distal to the splenic flexure, a left hemicolectomy or anterior rectal resection is option [16]. In high-risk patients with septic shock, diffuse fecal peritonitis, or in the presence of unresectable cancer, alternative interventions are right or subtotal colectomy with terminal ileostomy, Hartmann procedure, bowel bypass, or colostomy [18].  In colorectal cancer (CRC), obstruction and perforation may occur either alone or together, at the site of the neoplasm or proximally [19]. PAD is common cause of the small bowel obstruction (SBO) which may lead to the bowel strangulation, ischemia and perforation [19]. Small bowel perforation in our study was caused by strangulation and necrosis due to adhesive ileus in mostly cases.  Postoperative adhesive disease (PAD) is not an uncommon disorder, and the incidence ranges between 12% and 17% following abdominal surgery [20]. Postoperative course in perforative Crohn's disease is often characterized by post-operative complications, anastomotic leakage and recurrent disease [20]. We found Crohn's disease in 3 cases of small bowel perforation, etiology of the bowel perforation was conformed on postoperative histology. After bowel resection, there was no anastomotic complications in those patient. We did not follow-up this patient more than one month.  About 1–3% of patients with Crohn’s disease will present with a perforation of the intestine during their disease course [21]. In this study we found 5 causes of blunt trauma caused small bowel perforation. Four injuries occurred in a traffic accident and one patient fell from a tree. All patients recovered following surgery. Small bowel perforation caused by blunt abdominal trauma is rare but serious condition, because if the clinician does not have in mind this mechanism of bowel perforation, diagnosis may be delayed [24]. Patients with perforated gangrenous cholecystitis may have delays in presentation to the hospital, some of them presented with a benign course, with mild leukocytosis and a lack of peritoneal signs. [25] In case of complication, surgery and cholecystectomy is mandatory in 48-72 hours [26]. Mortality rates for gallbladder perforation decreased to 7- 16% in the following years owing to surgery, the developments in anesthesiology and intensive care conditions [28]. Laparoscopic approach is the treatment of choice for gallbladder disease, but an open approach and emergency laparotomy, cholecystectomy, lavage and drainage of the abdominal cavity is necessary in complicated cholecystitis with perforation [23]. Our study confirmed this attitude.  Perforation of the gallbladder is an infrequent complication of acute cholecystitis (2-15%), but it is associated with a high mortality rate without early treatment [30]. Reasons are delayed diagnosis and the irreversibility of bowel ischemia after a few hours. [30] Risk factors for this condition include age of >50 years, cardiovascular disorders, absolute arrhythmia, and presence of multiple comorbid conditions. The causes of this serious condition are: embolus of the superior mesenteric artery (50%), thrombosis (<25%), and rare by venous thrombosis. Patients with mesenteric ischemia present with abdominal pain and sometimes bloody diarrhea, but it is important to emphasize that clinical presentation, laboratory investigations and radiological finding may all be non-specific.[30] Treatment includes mesenteric revascularization and resection of necrotic bowel. Unfortunately, mortality and morbidity rates after bowel resection for mesenteric ischemia are high. [30] In our study we performed bowel resection and mesenteric revascularization in 2 patients. In others cases of AMI, all small intestines and right colon was necrotic, so operation was finished by exploration.  Acute mesenteric ischemia (AMI) is a life-threatening surgical emergency but a relatively uncommon cause of acute abdomen, because the mortality rates have a range of 45%–90%. [31]. The most common sites of intestinal perforation are the ileocecal and rectosigmoid regions. In our study peritonitis due to distal ileum perforation caused by foreign body was founded in 2 cases. Our approach was segmental resection of the terminal ileum and end to end anastomosis.  Accidental ingestion of a foreign body occurs rarely and intestinal perforation occurs in less than 1% of ingested bodies [32]. Clinical presentation is acute with signs of acute abdomen. Recommendation in acute abdomen with diffuse peritonitis is urgent laparotomy with drainage and salpingectomy [32]. Peritonitis secondary to bilateral pyosalpinx in a young healthy woman is not very common, in those cases infection my reached the peritoneal cavity through the Fallopian tubes [34]. The etiology of a hepatic abscess is complex, the infection my be cryptogenic in 69%, commonly in the setting of a comorbid illness such as diabetes mellitus , cardiopulmonary disease, malignancy, or liver cirrhosis [33]. Pyogenic liver abscesses may be caused by biliary tract disease, infection via the portal route associated with diverticulitis, pyelophlebitis, appendicitis, and proctitis; penetrating trauma and hepatic arterial microembolism in patients with sepsis [33]. In our study, we founded two cases of the ruptured pyogenic liver abscess in patients with diabetes mellitus, we performed debridement, pus evacuation and segment liver resection. Third patient had a perforated diverticulitis and huge liver abscess in the right lobe; surgery treatment was Hartmann procedure and liver resection. Rupture of a pyogenic liver abscess is relatively uncommon, reportedly occurring in 5.4% of cases [35]. Rupture of a pyonephrotic kidney is usually associated with a previous kidney abnormality, renal stones and, much less commonly, neoplasms [35]. We revealed intraoperatively the renal origin of peritonitis, kidney was complete destroyed in both patient, patients was in severe sepsis origin in ruptured pyonephrosis. Peritoneal fistulization of a pyonephrosis is an extremely rare event which invariably leads to generalized peritonitis [Even at the beginning of the new millennium generalized peritonitis still has a high mortality and morbidity. The atypical clinical presentation and delay in presentation can make diagnosis difficult. In addition to the availability of modern radiological methods, a careful clinical examination is still the most important way to achieve rapid, accurate diagnoses. Surgical attitude is rapid control contamination, abdominal lavage and abdominal drainage. In low risk patient, surgeons prefer a single definitive operation, but it is reasonable to use staged procedures and “second look" re-laparotomy in patients who are at high risk. There is no way to prevent peritonitis, however, efficient surgery, modern anesthesia and intensive care support my offer the chance of decreased morbidity and mortality of the intra-abdominal infections.


Considerable evidence indicates that the innate and adaptive immune systems participate in the recognition and destruction of cancer cells by a process known as cancer immunosurveillance. Tumor antigen-specific cytotoxic T-lymphocytes (CTL) are the major effectors in the immune response against tumor cells, and current approaches are essentially designed with the ultimate goal of inducing a strong CTL response. It is now obvious that some tumor cells can escape immunosurveillance, and accumulating evidence suggests such escape is tightly controlled by the tumor microenvironment, metabolic remodeling/hypoxia, cellular complexity and plasticity. As we develop a more complete view on the multifaceted role of tumor microenvironment in tumor development, progression, and in shaping tumor stroma, emerging observations now provide support for an essential role of tumor plasticity in resistance to CTL attacks. Along this line, epithelial mesenchymal transition (EMT) is an effective strategy by which cancer cells could gain plasticity. EMT is a transdifferentiation process whereby epithelial cells lose their epithelial properties while gaining mesenchymal properties . At leasIn our recent analyses, we found that silencing of WNT1-inducible signaling pathway protein 2 (thereafter WISP2) in MCF7 cells, resulted in EMT, coinciding with hyperactivity of TGF-β signaling, upregulation of stem cell factor KLF4, and impairment of CTL-mediated lysis [Recently various investigators pointed out the role of EMT in mounting resistance to anti-tumor immunity. Transduction of Snail in B16 melanoma cells resulted in inhibition of CTL lysis activity concomitantly with inhibition of dendritic cell maturation and expansion of suppressive Treg-like CD4+ Foxp3+ cells . In variWhile our knowledge is growing on the contribution of EMT and CSCs in cancer, it becomes clear that a better understanding of the pathways governing tumor plasticity will offer new therapeutic solutions in the future to combat cancer, predict response, and boost effectiveness of treatments. We argue that targeting carcinoma cell plasticity represents a novel strategy to better control the emergence of resistant variants. In this context, the design of innovative integrative immunotherapy approaches is warranted.


Neuroprotective therapies which focus on factors leading to retinal ganglion cells (RGCs) degeneration have been drawing more and more attention. The beneficial effects of nerve growth factor (NGF) on the glaucoma have been recently suggested, but its effects on eye tissue are complex and controversial in various studies. Recent clinical trials of systemically and topically administrated NGF demonstrate that NGF is effective in treating several ocular diseases, including glaucoma. NGF has two receptors named high affinity NGF tyrosine kinase receptor TrkA and low affinity receptor p75NTR. Both receptors exist in cells in retina like RGC (expressing TrkA) and glia cells (expressing p75NTR). NGF functions by binding to TrkA or p75NTR alone or both together. The binding of NGF to TrkA alone in RGC promotes RGC's survival and proliferation through activation of TrkA and several prosurvival pathways. In contrast, the binding of NGF to p75NTR leads to apoptosis although it also promotes survival in some cases. Binding of NGF to both TrkA and p75NTR at the same time leads to survival in which p75NTR functions as a TrkA helping receptor. This review discusses the current understanding of the NGF signaling in retina and the therapeutic implications in the treatment of glaucoma. Glaucoma is one of the leading causes of blindness worldwide. Glaucoma is characterized by retinal ganglion cell (RGC) degeneration and loss of visual field and it occurs with or without elevated intraocular pressure (IOP) . ApoptosNerve growth factor (NGF) was discovered in 1948. It prevents neuronal apoptosis in primary cultured neurons and reduces neuronal degeneration in animal models of neurodegenerative diseases , 6. In cβ, promotes wound healing, inhibits macrophage proliferation, and protects against nerve-injury-induced neuropathic pain [Growth factors are produced by our body and they have a very extensive role in the regulation of many cellular processes. The binding of growth factors to their receptors on the cell surface affects cellular survival, proliferation, and/or differentiation , 11. Forhic pain . in vitro and in vivo, have shown that NGF stimulates neurite outgrowth and axonal branching and extension [NGF is another important polypeptide growth factor which functions to regulate the growth and survival of nerve cells; it was discovered by Rita Levi-Montalcini and Stanley Cohen in the 1950s . NGF belxtension –20. Mostxtension .The clinical significance of NGF has been widely studied and it is recognized that NGF profoundly affects the development of both the young and the adult nervous systems . In the NGF plays its role by binding to its receptors located in the surface of cells. TrkA is the high affinity catalytically active receptor for NGF. NGF binding to TrkA leads to the phosphorylation of TrkA and activation of its downstream targets, such as protein kinase B (Akt) or extracellular signal-regulated protein kinase 1/2 (ERK1/2), which eventually cause neural differentiation and prevention of apoptosis . The othThe affinity of NGF binding to p75NTR receptor is weaker than NGF binding to TrkA, but the cell type distribution of p75NTR is wider than that of TrkA; TrkA receptor is mainly expressed at neurons responsive to NGF: peripheral sensory neurons, sympathetic neurons, and basal forebrain cholinergic neurons , while tTrkA is the high affinity catalytic receptor for the NGF and it mediates the main effects of NGF, which include cell growth, the formation and regeneration of neurites, and avoidance of programmed cell death . BindingPhosphorylation of the TrkA receptor leads to the interaction of TrkA and phosphatidylinositiol-3 kinase (PI3K). PI3K is activated and recruited to the plasma membrane and leads to the production of phosphoinositide 3,4,5-trisphosphate and membrane translocation of the serine/threonine-protein kinases Akt and Akt activation . Althougγ1) [γ1 supports activation of PKC signaling pathway and is thus involved in antimitogenic/mitogenic signaling.Another NGF-activated signaling pathway is the Ras-mediated activation of the mitogen-activated protein kinase (MAPK) pathway, which is initiated through recruitment and phosphorylation of Shc . Ras is γ1) . PLCγ1 s in vitro and in vivo. p75NTR activates Rac GTPase and activated c-jun N-terminal kinase (JNK), including an injury-specific isoform, JNK3 [κB (NF-κB) through p75NTR and during this process, p75NTR-mediated NF-κB activation enhances the survival response of developing sensory neurons to nerve growth factor [κB is a nuclear transcription factor that regulates expression of a large number of genes that are critical for the regulation of cell survival. Thus, in summary, the p75NTR receptor activates apoptotic signaling through the JNK cascade or cell survival through NF-κB pathway.The other receptor for NGF is p75NTR, but it is not a specific receptor for NGF as it also binds other neurotrophins, such as NT-3, NT-4/5, and BDNF . The rolrm, JNK3 . JNKs strm, JNK3 , so p75Nh factor . NF-κB i in vitro have shown that neurons coexpressing p75NTR and TrkA respond to lower concentrations of NGF [NGF binds to both TrkA and p75NTR receptors when they coexpressed on the outer cell membrane, even though the affinity of TrkA with NGF is much higher. Studiess of NGF , 57, whis of NGF . The ress of NGF . There is of NGF . These dGlaucoma is a severe eye disease, which is usually associated with absolute or relative elevated fluid pressure within the eyes and gradually progressive visual field loss. One major risk factor for glaucoma is the raised IOP. However, other mechanisms must be involved in the pathology of glaucoma because glaucoma can develop in the absence of elevated IOP and, in the clinic, different individuals respond differently to the elevated pressure . In someNotwithstanding the above, among the several causes for glaucoma, elevated eye pressure is one of the most important and best recognized acceptable risk factors . BesidesOptic nerve damage is a common optic neuropathy of glaucoma, which is characterized by loss of retinal ganglion cells . Both thIn normal conditions, primary cultured RGCs and transformed RGC-5 cells express various neurotrophins, such as brain-derived neurotrophic factor (BDNF), NGF, neurotrophin-3 (NT3), neurotrophin-4 (NT4), and also their relevant receptors, such as TrkA, p75NTR, or TrkB. RGC-5 cells also secrete NT3, BDNF, NGF, and NT4 into the cultured media . In the While, in pathological conditions, such as glaucoma, the increased IOP will alter the level of NGF and NGF receptor expression, glaucoma significantly reduces the content of NGF in the cerebrospinal fluid (CSF) and lateral geniculate nucleus (LGN), but serum NGF protein levels may not be affected; this finding suggests that the NGF present in the CSF is most likely taken up by damaged retinal or brain neurons. Ongoing research shows that glaucoma increases the basal level of TrkA in the LGN and NGF administration further enhances this increase. However glaucoma had no effect on the expression of p75NTR at early stage, while NGF still enhanced the level of p75NTR; these results support the contention that glaucoma altered the basal level of NGF and NGF receptors in brain visual centers . After 2In the CNS, NGF possesses obvious effects on modulating neuronal inputs and thereby synaptic plasticity. NGF augmentation significantly enhanced cholinergic neuronal markers and facilitated induction of hippocampal long-term potentiation (LTP); moreover, blockade of endogenous NGF significantly reduced hippocampal LTP and impaired retention of spatial memory . These fOptineurin (OPTN) and myocilin (MYOC) are two genes linked to glaucoma . Rezaie κB and JNK activation, whereas TrkA mediates MAPK and Akt activation and when the two receptors are expressed together, TrkA blocks the p75NTR-mediated JNK activation, but NF-κB is unaffected [As discussed above, the role of the receptors, TrkA and p75NTR, is not consistent in the CNS. It is widely accepted that sympathetic neuron survival is regulated positively by TrkA and negatively by p75NTR. The apoptotic effect of p75NTR signaling occurs only under suboptimal survival conditions, while TrkA-mediated responses occur when survival conditions are optimal –91. In faffected , 90. in vitro studies, in vivo studies indicate that NGF binds to TrkA and p75NTR but fails to promote the survival of axotomized RGCs in the adult retina; however, TrkA agonists showed robust neuroprotective effects [ in vivo [In the optical system, the two receptors may also function differently. In normal adult retinas, TrkA is expressed in RGCs, whereas P75 is expressed in glia. In contrast to results from effects . Pharmac in vivo . Treatme in vivo . In a mo in vivo . In summ in vivo  and p75N in vivo . Thus, iNGF is a widely studied growth factor which possesses significant neuroprotective effects on neurons against various insults. In the central nervous system, NGF deficiency is involved in age-related neurodegenerative diseases; a deficit in the signaling and/or transport of NGF also leads to neurodegeneration . Treatme ex vivo gene delivery or biologically stable small molecules that could bind and activate the TrkA signaling pathway are alternative strategies [Sustained elevated IOP causes RGC degeneration and results in cell death. In animal models, glaucoma is induced by the injection of hypertonic saline into the episcleral vein of the eyes. A rat model of glaucoma showed that glaucoma led to progressive degeneration of RGCs, with the loss of nearly 40% of these cells after 7 weeks of treatment. The RGC loss is associated with the downregulation of NGF and NGF receptor expression in the retina and ocular treatment with NGF significantly reduced the deficit induced by glaucoma . Anotherrategies , 102. Frrategies ; howeverThe ultimate goal of glaucoma research is to find new compounds that will not only normalize IOP but also arrest or even reverse apoptotic damage to the optic nerve and RGCs and slow the rate of progression of the disease. This review discussed current knowledge of glaucoma, with an emphasis on the NGF and NGF receptor(s) signaling pathways, and further explored the possibility of targeting the NGF signaling pathway as a strategy for the treatment of glaucoma. NGF offers the promise of actually restoring visual function through acting on the TrkA receptor; however, we should be cautious regarding the future of NGF-dependent treatments in the armamentarium of glaucoma therapy as most of the present studies were in animal models and none has reached clinical success. After more research and deeper mechanistic understanding a randomized, controlled glaucoma, clinical trials need to be performed to evaluate the therapeutic effect of NGF in the treatment of glaucoma and this also will involve an evaluation of the magnitude and occurrence rate of adverse effects. In the future, developments of novel pharmacological interventions for glaucoma may include the development of small molecules that are specific for TrkA receptors in the optical nerves. With protein chemistry entering into a new phase of “peptide design,” it is hoped that careful design of these small molecules may be able to limit biological activity to only the wanted effects and minimize the adverse effects. Other strategies such as chemical immobilization of NGF into or onto the matrix and physical encapsulation of NGF in the delivery system may also be promising in the future studies. Such novel ideas for trophic support are still developing and may provide future “smart drugs” for glaucoma.


Neurotransmitters/neuromodulators binding to metabotropic receptors are essential to chemotransmission in the CB, and cAMP is central to this process. cAMP also contributes to raise intracellular Ca2+ levels, and is intimately related to the cellular energetic status (AMP/ATP ratio). Furthermore, cAMP signaling is a target of multiple current pharmacological agents used in clinical practice. This review (1) provides an outline on the classical view of the cAMP-signaling pathway in the CB that originally supported its role in the O2/CO2 sensing mechanism, (2) presents recent evidence on CB cAMP neuromodulation and (3) discusses how CB activity is affected by current clinical therapies that modify cAMP-signaling, namely dopaminergic drugs, caffeine (modulation of A2A/A2B receptors) and roflumilast (PDE4 inhibitors). cAMP is key to any process that involves metabotropic receptors and the intracellular pathways involved in CB disease states are likely to involve this classical second messenger. Research examining the potential modification of cAMP levels and/or interactions with molecules associated with CB hyperactivity is currently in its beginning and this review will open doors for future explorations.Chronic carotid body (CB) activation is now recognized as being essential in the development of hypertension and promoting insulin resistance; thus, it is imperative to characterize the chemotransduction mechanisms of this organ in order to modulate its activity and improve patient outcomes. For several years, and although controversial, cyclic adenosine monophosphate (cAMP) was considered an important player in initiating the activation of the CB. However, its relevance was partially displaced in the 90s by the emerging role of the mitochondria and molecules such as AMP-activated protein kinase and O Highly specialized chemosensory type I cells of the mammalian carotid bodies (CBs) sense acute changes in PaO2, PaCO2 and pH, and, upon stimulation, release neurotransmitters (NTs) that either inhibit or activate chemosensory fibers projecting into the central nervous system (CNS). The functional consequence of CB stimulation is the initiation of important cardiovascular, respiratory and metabolic reflexes. These reflexes include an increase in minute ventilation, a sympathetically mediated elevation in heart rate and peripheral vasoconstriction and an augmentation in adrenaline release from the adrenal medulla, with the latter leading to an increase in arterial blood glucose concentration.Adequate homeostatic regulation of arterial oxygen  and respective receptors in the CB has increased considerably over the last 20 years  neurons and CB type I and type II cells is another source of NTs/NMs release in the CB. PG neurons present catecholaminergic traits  exerts inhibitory effects on the CB in most species, except rabbit,  is a common downstream signaling molecule of numerous receptors expressed in the type I cells, and is coupled to cellular energetic status (AMP/ATP ratio). This article therefore aims to summarize how changes in CB cAMP levels in physiology, pathology and following pharmacological intervention may be central to alterations in type I cell excitability leading to chemoafferent discharge and cardiorespiratory and metabolic reflex responses.2  was originally prompted by the identification of secreted NTs and their receptor-mediated effects on CB chemoreceptor responses Table . These sFitzgerald and co-workers, were the first to identify cAMP in the CB cat homogenates  preparations) and cAMP detection methods  have all been credited for the discrepancies reported in the literature regarding the relevance of cAMP signaling in CB function , and by the fluctuating activity of downstream effectors  and G-inhibitory (Gi) proteins as well as by CO2 and HCO−3 (depending on specific AC isoforms- see below). [cAMP]i can also be modified by direct diffusion from one cell to another through gap junctions  and the more recently described soluble adenylyl cyclase (sAC)  express tmAC1, tmAC2, tmAC3, tmAC4, tmAC6 and tmAC9, with tmAC1, tmAC4 and tmAC6 exhibiting the highest fold expression level  to 44mM/10% (isohydric hypercapnia) does not alter cAMP content, PKA activity or CSN discharge frequency and, under these conditions, these assays were insensitive to the sAC inhibitor KH7  and ion channels. AKAPs tether PKA to particular cellular organelles and to the plasma membrane confining the PKA signaling to a small pool within the cells   diminishes the rise in L-type CaDifferent effects of PDE4 inhibitors on cAMP accumulation induced by hypoxia  mediated phosphorylation can modify PDE4 specific isoforms activity and subsequently alter the sensitivity to selective inhibitors  for the RasGTPase homologs, Rap1 and Rap2. EPAC is composed of two regions: an N-terminal regulatory region containing a cAMP-binding site and a C-terminal catalytic region, with GEF activity  and Gq signaling pathways within the type I cell likely occurs. Better characterization of this interaction could be particularly insightful given the known upregulation of Gq signaling associated with CB dysfunction in sleep disorder breathing  and inhibitor (brefeldin) on the release of CAs  and CaM-dependent kinases and, in turn, modulate cAMP production by regulating activity of AC and PDE. Since CNGC and HCNC are also permeable to Na+ and K+, they can also alter the membrane potential in electrically active cells. The presence of these channels in the rat CB has been suggested by the work of Stea and co-workers  and hyperpolarization-activated cyclic nucleotide-modulated (HCNC) ion channels. These channels belong to a superfamily of voltage-gated cation channels, and thus the binding of cAMP to these channels is translated into changes in membrane potential and influx of Ca2A and A2B Ado receptors .Observations from our laboratory show that cAMP levels are higher during normoxia than in hypoxic or hyperoxic superfusate in the whole CB from young and adult rats  on peripheral chemoreflexes is unknown but beneficial effects of loxapine on agitation and breathing patterns during weaning from mechanic ventilation have been described  is clinically useful to revert paroxysmal supraventricular tachycardia and causes dyspnea and chest discomfort mediated by CB activation  and the European Medicines Agency (EMA) for the treatment of severe chronic obstructive pulmonary disease (COPD), due to its anti-inflammatory and bronchodilator effects. No evidences of roflumilast effects on CB activity have been reported and those are difficult to address essentially because of their CNS effects and the variability of the PDE inhibitors efficacy in hypoxic conditions  (Whipp and Ward, 2/CO2 related diseases. However, manipulation of cAMP signaling may have consequences in the CB, that are clinically relevant and that have not yet been identified.From the above evidence, one can conclude that the manipulation of cAMP signaling pathway is important to address O2 chemotransduction to a central signaling molecule that is converged upon by multiple NTs/NMs, which collectively maintain an equilibrated CB activity. It remains to be seen whether modification of cAMP can improve patient outcomes in diseases associated with CB impairment or hyperactivity. We suggest that cAMP has an important role in the homeostasis of the CB since cAMP levels seem to be higher under normoxic conditions. Despite the increase in knowledge of CB physiology, the activity of tyrosine hydroxylase is still the hallmark of the CB and cAMP the classical second messenger of dopamine D2 receptor signaling.The importance of cAMP to CB physiology has moved from a discarded player in the OUnderstanding how calcium and cAMP cooperate in dysfunction CB, will be fundamental to understand the role of these second messengers in the CB transduction mechanism. Additionally, systemic pharmacological manipulation of cAMP signaling can have clinically relevant consequences mediated by the CB. This may proven to be an exciting field of research that is still currently unexplored.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.


Some subcellular compartments are more oxidizing (such as lysosomes or peroxisomes) whereas others are more reducing . Moreover, although more reducing, mitochondria are especially susceptible to oxidation, most likely due to the high number of exposed thiols present in that compartment. Recent advances in the development of redox probes allow specific measurement of defined ROS in different cellular compartments in intact living cells or organisms. The availability of these tools now allows simultaneous spatio-temporal measurements and correlation between ROS generation and organelle and/or cellular function. The study of ROS compartmentalization and microdomains will help elucidate their role in physiology and disease. Here we will examine redox probes currently available and how ROS generation may vary between subcellular compartments. Furthermore, we will discuss ROS compartmentalization in physiological and pathological conditions focusing our attention on mitochondria, since their vulnerability to oxidative stress is likely at the basis of several diseases.Reactive oxygen species (ROS) formation and signaling are of major importance and regulate a number of processes in physiological conditions. A disruption in redox status regulation, however, has been associated with numerous pathological conditions. In recent years it has become increasingly clear that oxidative and reductive modifications are confined in a spatio-temporal manner. This makes ROS signaling similar to that of Ca Reactive oxygen species (ROS) formation and redox signaling are well known to play a major role in physiology as well as in a variety of pathologies. For instance, in the heart, cardiomyocyte differentiation, and excitation-contraction coupling are under tight redox control  or with another protein thiol  are maintained at stable values and are not in equilibrium relative to each other in different organelles function, it is necessary to define which species are produced, in what amount and to characterize them in a spatio-temporal manner. Redox potential of a specific compartment or cell can be studied using a variety of techniques to identify and quantify major redox couples or redox sensitive proteins within organelles. For instance, high-performance liquid chromatography (HPLC) is used for the quantification of GSH/GSSG and NAD(P)H/NAD(P)s Jones, , whereass Jones, . Althougs Jones, . More reAlthough several small molecule fluorescent probes are available, only a few of them can be targeted to specific subcellular compartments Table . It shouMitoSOX Red is widely used for measurement of superoxide formation in the mitochondria of live cells  -based redox sensitive proteins were developed introducing cysteine residues onto fluorescent protein scaffolds . Initially, roGFP1 was developed that provided ratiometric fluorescence readout and increased sensitivity compared to rxYFP , ER, nucleus, lysosomes, and endosomes  and responds to either GSH or GSSG in a time scale of minutes and senses redox potential changes between −240 and −320 mV  into the regulatory domain of E. coli H2O2 sensing protein OxyR  based redox sensors. Initially, a linker containing two cysteine residues was placed between enhanced cyan and yellow fluorescent protein FRET pair, but showed a too small dynamic range of the probes to be used for imaging  were developed placing circularly permuted fluorescent proteins (cpFPs) into a tandem dimer of bacterial Rex protein, capable of binding NADH  photochemical properties of the fluorescent protein, (2) its concentration and subcellular localization, (3) the excitation intensity. Importantly, the total excitation light dose should be kept to minimum and it is preferable to use probes with longer wavelength excitation light, since excitation at a shorter wavelength is more damaging to cells because of increased efficiency of ROS production  among others, are prominent sites for ROS generation , rapid mitochondrial depolarization, and increase in pH. These changes were independent of mitochondrial Ca2+ levels, but highly dependent on mitochondrial ROS formation and were increased exposing axons to higher neuronal activity. This suggests that these reversible mitochondrial redox changes might serve as a signal to overcome acute challenges and protect from eventual damage. However, under pathological conditions, such as in a chronic amyotrophic lateral sclerosis model or axotomy, rapid mitochondrial Ca2+ increase led to long-lasting mitochondrial oxidation, irreversible changes in shape and opening of the permeability transition pore (PTP), suggesting that noxious levels of stress may induce more permanent, Ca2+-induced, mitochondrial derangements.A recent study examining the fine line between mitochondrial redox signals in physiology and pathology was performed on mice expressing Grx1-roGFP2 sensor in neuronal mitochondria , triggered by transient openings of the mitochondrial PTP , activated by hypoxia, is suggested to respond to and modulate mitochondrial oxidant production, through transcriptional regulation of several microRNAs that control the expression of components of the electron transport chain, lactate dehydrogenase A and PDK1 (pyruvate dehydrogenase kinase 1) . In addition, mitochondrial ROS formation through MAOs was shown to lead to the oxidation of myofibrillar proteins in the failing heart, an event that negatively correlated with cardiac function  and extracellular signal-regulated kinases (ERK1/2) activation in the cytosol, but without affecting cytoplasmic GSH and Trx1 redox state  and NF-E2-related factor 2 (Nrf2)  are maintained at more reduced values than the ones in the cytoplasm, protecting the genome from ROS-induced damage. Indeed, many studies found oxidation in mitochondria and cytoplasm, but were unable to detect any changes in nuclei exposed to oxidants , endosomes and the ER membrane allows visualization of sites of focal ROS formation within compartments  on the endosomes and the cytoplasmic side of the ER membrane, but no H2O2 formation was observed associated with the plasma membrane. PDGFR activation instead generated H2O2 microdomains at the plasma membrane and only after prolonged incubation on the endosomes, suggesting that a specific plasma membrane-residing Nox pool was activated  contain cysteine residues critical for enzyme activity, whose oxidation leads to their reversible inactivation Tonks, . The pre+ resulted in oxidation in the mitochondrial matrix that preceded the one in the cytosol, but did not lead to oxidative stress in the IMS  it allows for better characterization of ROS signals in compartments and microdomains, and (2) provides an explanation for antioxidants failure in treating several pathologies. The unsuccessful outcome of several clinical trials evaluating the therapeutic potential of antioxidants for the treatment of heart failure raised questions regarding the importance of ROS or oxidative stress. Taking into account data available and given the importance of ROS both in physiology and pathology, we propose that inhibition of specific processes that generate ROS rather than general antioxidants administration may prove as a successful therapeutic strategy.Nina Kaludercic and Fabio Di Lisa have a pending patent application entitled “Treatment of heart failure and associated conditions by administration of monoamine oxidase inhibitors,” number: 20090286883. 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.


Drosophila and mammals. The nuclear periphery is one of the nuclear compartments implicated in gene regulation. It consists of the nuclear envelope (NE) and the nuclear pore complexes (NPC), which have distinct roles in the control of gene expression. The NPC has recently been shown to tether proteins involved in the sumoylation pathway. Here, we will focus on the importance of gene positioning and NPC-linked sumoylation/desumoylation in transcription regulation. We will mainly discuss observations made in the yeast Saccharomyces cerevisiae model system and highlight potential parallels in metazoan species.Increasing evidence indicates that besides promoters, enhancers, and epigenetic modifications, nuclear organization is another parameter contributing to optimal control of gene expression. Although differences between species exist, the influence of gene positioning on expression seems to be a conserved feature from yeast to  It is well established that during interphase, mammalian chromosomes occupy distinct nuclear regions called chromosome territories  was proposed to act as a DNA zip code that is both necessary and sufficient to target a gene to the NPC -inducible genes  in higher eukaryotes  evolutionarily conserved polypeptide SUMO  on lysine residues. In SUMO proteomics studies indicate that most sumoylated proteins are nuclear , which involves an initial derepression step in order to achieve optimal coactivator recruitment. Derepression consists of extensive chromatin remodeling and the dissociation of the repressor Mig1 from the promoter, a process induced by phosphorylation of Mig1 by the Snf1 kinase , which mainly relies on the recruitment of the SAGA coactivator complex by the Gal4 activator , and Sec13 (component of Nup107/160 complex homologous to yeast Nup84C), have been implicated in transcription regulation , Ulp1 has been shown to relocate to the septin ring in mitosis to regulate septin desumoylation , further reinforcing the links between the sumoylation of a chromatin regulator and gene expression.A number of chromatin-modifying enzymes are sumoylated, and the modification is usually linked to repression   may also influence their interaction with these histone-modifying enzymes Fig. .An emerging view is that a multitude of nuclear proteins are either sumoylated and/or contain SIM domains, which facilitate the formation of vast protein networks that contribute to the constitution of nuclear subdomains enriched in specific factors important for optimal regulation of gene expression (Hickey et al. While we focused on the positive effect of desumoylation on gene expression, transcription factors such as Gcn4 become sumoylated during activation. In this case, sumoylation promotes ubiquitination and degradation of Gcn4 by the proteasome, favoring the rapid on-off switch of gene expression (Rosonina et al. Importantly, besides transcription and mRNA biogenesis, NPC-linked SUMO metabolism also contributes to genome stability. Indeed, both Ulp1 and the STUbL Slx5/Slx8 have been implicated in DNA repair and telomere maintenance (Nagai et al.


The immune response of the CNS is a defense mechanism activated upon injury to initiate repair mechanisms while chronic over-activation of the CNS immune system (termed neuroinflammation) may exacerbate injury. The latter is implicated in a variety of neurological and neurodegenerative disorders such as Alzheimer and Parkinson diseases, amyotrophic lateral sclerosis, multiple sclerosis, traumatic brain injury, HIV dementia, and prion diseases. Cyclooxygenases (COX-1 and COX-2), which are key enzymes in the conversion of arachidonic acid into bioactive prostanoids, play a central role in the inflammatory cascade. J2 prostaglandins are endogenous toxic products of cyclooxygenases, and because their levels are significantly increased upon brain injury, they are actively involved in neuronal dysfunction induced by pro-inflammatory stimuli. In this review, we highlight the mechanisms by which J2 prostaglandins (1) exert their actions, (2) potentially contribute to the transition from acute to chronic inflammation and to the spreading of neuropathology, (3) disturb the ubiquitin-proteasome pathway and mitochondrial function, and (4) contribute to neurodegenerative disorders such as Alzheimer and Parkinson diseases, and amyotrophic lateral sclerosis, as well as stroke, traumatic brain injury (TBI), and demyelination in Krabbe disease. We conclude by discussing the therapeutic potential of targeting the J2 prostaglandin pathway to prevent/delay neurodegeneration associated with neuroinflammation. In this context, we suggest a shift from the traditional view that cyclooxygenases are the most appropriate targets to treat neuroinflammation, to the notion that J2 prostaglandin pathways and other neurotoxic prostaglandins downstream from cyclooxygenases, would offer significant benefits as more effective therapeutic targets to treat chronic neurodegenerative diseases, while minimizing adverse side effects. Chronic neuroinflammation is recognized as a primary mechanism involved in the pathogenesis of a variety of neurodegenerative disorders including Alzheimer, Parkinson, and Huntington diseases as well as amyotrophic lateral sclerosis  are a family of 20-carbon unsaturated fatty acids produced via the cyclooxygenase pathway in response to numerous extrinsic and intrinsic stimuli Figure . PLA2 isCyclooxygenases, which are bifunctional enzymes inserted into the ER and nuclear membranes, will then catalyze the cyclooxygenation of arachidonic acid to PGG2 followed by hydroperoxidation of PGG2 to PGH2 .PGJ2 and its metabolites are not stored in tissues or cells and their production increases with diverse stimuli. Prostaglandins are largely produced in the brain by activated microglia, reactive astrocytes and neurons. During CNS inflammation, these cells make large quantities of prostaglandins such as PGE2 and PGD2 , and therefore reported levels of free J2 prostaglandins do not represent their total amounts.J2 prostaglandins Table  are bioaDespite their lipid nature, prostaglandins are charged anions thus have low intrinsic permeability across the plasma membrane, but they cross it twice Figure : once foProstaglandins also can be transferred from cell to cell via exosomes, which are extracellular bioactive vesicles released from multivesicular bodies that mediate intercellular signaling , to which they bind with high affinity  by endogenous compounds such as PGJ2 is regarded as playing an important role in determining whether neurons will live or die  of the DP2 receptor  PK11195  -propyl]piperidine], suppressed astroglyosis, demyelination, twitching and spasticity. These results support the notion that PGD2 and perhaps its metabolites, are key to the pathological demyelination occurring in the twitcher mouse, and the neuroprotective potential of manipulating the PGD2-signaling to overcome demyelination in Krabbe disease  Table . MAGL, win vitro and in vivo  were developed to specifically deliver prostaglandins, in this case PGA2, to injured arterial wall cells of atherosclerotic mice (Homem de Bittencourt et al., In conclusion, much remains to be discovered about the biology of the J2 prostaglandins, to prevent their neurotoxic effects and possibly “trick” ongoing inflammation into resolution. Further investigations may provide important clues “to bring us into a new era of inflammation research, which, if approached with creativity and persistence, might provide numerous benefits for those suffering from inflammation-mediated diseases” Gilroy, .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.


Heme oxygenase (Hmox) is an endogenous system that offers protection against placental cytotoxic damage associated with preeclampsia. The Hmox1/carbon monoxide (CO) pathway inhibits soluble Flt-1 (sFlt-1) and soluble Endoglin (sEng). More importantly, statins induce Hmox1 and suppress the release of sFlt-1 and sEng; thus, statins and Hmox1 activators are potential novel therapeutic agents for treating preeclampsia. The contribution of the Hmox system to the pathogenesis of preeclampsia has been further indicated by the incidence of preeclampsia being reduced by a third in smokers, who had reduced levels of circulating sFlt-1. Interestingly, preeclamptic women exhale less CO compared with women with healthy pregnancies. Hmox1 is reduced prior to the increase in sFlt-1 as Hmox1 mRNA expression in the trophoblast is decreased in the first trimester in women who go on to develop preeclampsia. Induction of Hmox1 or exposure to CO or bilirubin has been shown to inhibit the release of sFlt-1 and sEng in animal models of preeclampsia. The functional benefit of statins and Hmox1 induction in women with preeclampsia is valid not only because they inhibit sFlt-1 release, but also because statins and Hmox1 are associated with anti-apoptotic, anti-inflammatory, and anti-oxidant properties. The StAmP trial is the first randomized control trial (RCT) evaluating the use of pravastatin to ameliorate severe preeclampsia. This proof-of-concept study will pave the way for future global RCT, the success of which will greatly contribute to achieving the United Nations Millennium Development Goals (MDG4 and MDG5) and offering an affordable and easily accessible therapy for preeclampsia. If left untreated it can lead to debilitating and potentially eclamptic seizures that can cause coma and even death. According to the Preeclampsia Foundation, globally preeclampsia accounts for 76,000 maternal deaths and 500,000 infant deaths every year. Currently, the lack of effective pharmacological options to treat preeclampsia indicates that there is an unmet need for an affordable and easily accessible treatment to prevent maternal and infant mortality. Factors originating in the placenta are likely to be responsible for the condition, yet the exact etiology of the disorder is unknown. Currently, the only curative management strategy is the premature termination of pregnancy and the delivery of the placenta. Compelling data have pointed to maternal endothelial dysfunction as the central phenomenon responsible for the clinical signs of the disorder–hypertension and proteinuria    has led in vitro removal of sFlt-1 from the placental condition media of preeclamptic women by immunoprecipitation restored the angiogenic imbalance and promoted the capillary-like tube formation similar to normal pregnancy  is known to maintain endothelial cell integrity . The antregnancy . Furtherregnancy . Clinicaregnancy . Collect3Heme oxygenase (Hmox) is the rate-limiting enzyme responsible for the degradation of heme in the endoplasmic reticulum to generate an equimolar amount of biliverdin, free iron, and carbon monoxide (CO) . The patvia its products , which regulate important biological processes including oxidative stress, inflammation, apoptosis, and angiogenesis in various conditions , the Hmox system is able to provide mechanisms for protection during pregnancy against a variety of physiological threats. Hemoproteins are responsible for the transfer of oxygen from the mother to the fetus during pregnancy and heme is produced by the trophoblast, the site of exchange of substances between maternal blood and fetal circulation. The placental vascular endothelium and syncytiotrophoblasts are exposed to high concentrations of hemoglobin owing to their direct contact with fetal and maternal blood. As has been previously demonstrated, hemoglobin and free heme can undergo auto-oxidation to produce superoxide and hydrogen peroxide, which in turn promote the formation of reactive oxygen species (ROS) and damaging free radicals . BecauseThe syncytiotrophoblast is also directly exposed to maternal blood and to maternal immunological attack and other inflammatory stressors such as tumor necrosis factor-α (TNF-α), interferon-γ, and interleukins. These inflammatory mediators and other stressors may induce the expression of Hmox1 in the syncytiotrophoblasts to counteract the injurious effect of inflammation on the placenta and to prevent oxidative injury. Indeed, the pharmacological induction of Hmox1 in human placental villous explants was shown to offer placental cytoprotection . In pree3.3In recent years, the functional benefit of Hmox in preeclampsia has gained increasing importance, in particular, after the publication demonstrating that the adenoviral overexpression of Hmox-1 or direct exposure to CO reduce both basal and VEGF-E-stimulated sFlt-1 release from human umbilical vein endothelial cells (HUVEC), while siRNA-mediated Hmox1 knockdown increases sFlt-1 release . Another+/−) resulted in Hmox1+/− placentas that had significantly lower Hmox1 mRNA and protein levels compared with wild-type littermates. Additionally, the Hmox1+/− placentas were smaller than the wild-type placentas. More importantly, the diastolic blood pressure and plasma sFlt-1 level were significantly elevated in pregnant Hmox1+/− mice compared with their pregnant wild-type counterparts (Other laboratories have used mice models to demonstrate the effect of the loss of function of the Hmox1 system on pregnancy. Crossbreds of Hmox1 heterozygote mice (Hmox1terparts . A recenterparts . These sterparts .The reduced uterine perfusion pressure (RUPP) hypertension model, in which chronic RUPP leads to endothelial dysfunction and hypertension in the pregnant rat, has also been used as a tool to study the preeclamptic syndrome. Gilbert and colleagues demonstrated that the expression of Hmox1 in the placenta of the RUPP pregnant rats during placental ischemia was significantly reduced compared with control non-ischemic rat placentas. Interestingly, elevation in both circulating sEng and placental expression of sEng was observed in these pregnant rats . To subs1-AA) from preeclamptic women injected into pregnant mice mediated the release of TNF-α, which subsequently induced the release of circulating sEng and sFlt-1 to impair placental angiogenesis. Of importance, this study also confirms that the pharmacological induction of Hmox1 by hemin prevented TNF-α from inducing sEng production in mice and that it also blocked the ability of serum obtained from pregnant mice injected with preeclamptic IgG to stimulate sEng production by human endothelial cells  . The maiStatins are primarily used to treat hypercholesterolemia by inhibiting the enzyme HMG-CoA reductase in the liver, thereby lowering LDL cholesterol production . They al2S) and the H2S-producing enzyme cystathionine γ-lyase (Cth) may account for the reduction in PlGF in preeclampsia  and because of a small case series on the teratogenic effects of the original statins in use at the time . Howeverde novo synthesis (fetus or placenta) rather than from maternal sources. Fetal mutations in the cholesterol synthesis pathway do not synthesize cholesterol and have very low concentrations of cholesterol in their blood despite normal maternal levels, suggesting a minor role of maternal exogenous sources (The majority of fetal cholesterol originates from  sources . Thus, aApart from inhibiting the release of sFlt-1 , statinsIn order to develop other therapies apart from statins to tackle preeclampsia, basic research is still needed to elucidate fully the role of Hmox1 and its metabolites. Murine models do not equate to preeclampsia in women, but they can replicate many of the preeclampsia symptoms and thus offer very useful tools for performing proof of principle experiments to determine the role of specific genes and therapies. What is now needed is the use of sophisticated knockout and tissue-specific knockdown mouse models combined with further clinically based studies, as has been the case in the cardiovascular field.


The intratumoral metabolism and synthesis of estrogens, as a result of the interactions of various enzymes, is more important than blood uptake to maintain mammary gland estrogen levels in menopausal females. Additionally, estrogens are considered to play an important role in the pathogenesis and development of hormone-dependent breast carcinoma. Paracrine interactions among malignant epithelial cells and proximal adipose and endothelial cells, through cytokines and growth factors produced by breast tumor cells, modulate estrogen production at the mammary tumor level and, as a consequence, the genesis and development of mammary tumors. The aim of the present review is to summarize the recent findings describing the mechanisms by which melatonin is able to modulate the crosstalk among malignant epithelial, endothelial and adipose cells in breast cancer.Melatonin, the main secretory product of the pineal gland, is an oncostatic agent that reduces the growth and development of various types of tumors, particularly mammary tumors whose growth is dependent on estrogens. Previous  At the 2 (PGE2) is an important regulator of aromatase gene expression via promoters II and I.3 (2 occurs through the activity of the cyclooxygenases (COXs), rate-limiting enzymes that catalyze the conversion of arachidonic acid to prostaglandins. Promoter I.3 and II are considered to be the major promoters driving aromatase expression in breast cancer and surrounding adipose tissue. One of the mechanisms through which melatonin modulates aromatase enzyme in breast tumor cells is through its downregulatory action on the expression of COX enzymes, COX-1 and COX-2, which decrease the levels of PGE2. Lower levels of PGE2 result in decreased intracellular levels of cAMP, which in turn diminish the activation of promoters I.3 and II, and result in decreased aromatase expression  animals. The stimulatory effects of tumor development induced by testosterone , due to the aromatase action, or estrone sulfate , are suppressed by the administration of melatonin. Tumors from animals treated with melatonin have the lowest microsomal aromatase and sulfatase activity , interleukin 6 (IL-6) and IL-11, which are upregulated by estrogens. These cytokines inhibit the differentiation of surrounding fibroblasts into mature adipocytes, through the selective inhibition of expression of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα), and also stimulate aromatase expression in these undifferentiated fibroblasts  19,34,3,334,35. in vitro the situation occurring in the mammary tumor is to use cocultures of malignant epithelial cells with fibroblasts or endothelial cells. The presence of malignant epithelial cells in the cocultures inhibits the differentiation of preadipocytes to adipocytes and reduces the intracytoplasmic triglyceride accumulation, an indicator of adipogenic differentiation. The presence of malignant cells also stimulates the aromatase activity in the fibroblasts. Melatonin counteracts the inhibitory effect on adipocyte differentiation induced by malignant epithelial cells, and also counteracts the stimulatory effect of the presence of breast cancer cells on aromatase activity in fibroblasts ..45).in vitro the paracrine interaction between these cells in the mammary tumors. The presence of malignant epithelial cells in the cocultures is able to stimulate the endothelial cell proliferation and increase the VEGF levels in the culture media. Melatonin counteracts the stimulatory effects on endothelial cell proliferation and on VEGF protein levels in the coculture media. The changes in endothelial cell proliferation induced by melatonin are mediated by an inhibition of the synthesis of VEGF in malignant epithelial cells. Conditioned media from malignant cells stimulate endothelial cell proliferation, and this effect is significantly counteracted by anti-VEGF and melatonin (VEGF, a major regulator of endothelial growth, added to endothelial cell cultures stimulates the proliferation of these cells and melatonin counteracts this effect . Melatonelatonin .All these findings suggest that melatonin may play a role in the paracrine interactions between malignant epithelial cells and proximal endothelial cells, through a downregulatory action on VEGF expression in human breast cancer cells, which decreases the levels of VEGF surrounding endothelial cells. Lower levels of VEGF may be important in reducing the number of estrogen-producing cells proximal to malignant cells, as well as in decreasing tumoral angiogenesis. Antiangiogenic activity of melatonin against the pro-angiogenic effects of breast cancer cells has also been described . Recentl2 (2 synthesis induced by VEGF may directly promote angiogenesis and melatonin through its downregulatory action on the expression of COX enzymes, which decrease the levels of PGE2 and reduce angiogenesis. On the other hand, melatonin inhibits aromatase activity and expression in endothelial cells by regulating gene expression of specific aromatase promoter regions, thereby reducing the local production of estrogens (Melatonin may play a role in the paracrine interactions that take place between malignant epithelial cells and proximal endothelial cells, acting by different mechanisms. On one hand, melatonin exerts antiangiogenic effects and may be important in reducing endothelial cell proliferation, invasion, migration and tube formation, through a downregulatory action on VEGF and PGE2 . PGE2 systrogens .2 by malignant cells, which upregulates aromatase expression both in the tumor itself and in the surrounding adipose tissue and enhances the production of IL-11 by tumor cells. Endothelial cells also produce estrogens from androgens precursors. Melatonin decreases the activation of promoter I.7 and results in decreased aromatase expression. In addition, melatonin reduces endothelial cell proliferation, invasion, migration and tube formation, through a downregulatory action on VEGF. This melatonin modulation of epithelial-stromal interactions favors lower numbers of undifferentiated fibroblasts, angiogenesis and reduced local estrogen concentrations in breast tumors.Several lines of evidence highlight the contribution of the tumor microenvironment to its growth and maintenance. Cells immediately adjacent to the tumor are not only passive structural support but also active elements in tumor progression. Among the numerous different cell types surrounding breast cancer cells, the most abundant are those that compose mammary adipose tissue. Ninety percent of these resident cells of adipose tissue are fibroblasts, the precursors of mature adipocytes, and 7% are endothelial cells . EpithelMelatonin may play a role in the paracrine interactions that occur between malignant epithelial cells and proximal adipose and endothelial cells, through a downregulatory action on cytokines and growth factors produced by breast tumor cells. The actions of melatonin described in the present review involve antiproliferative, antiaromatase and antiangiogenic effects, and suggest that melatonin may potentially be beneficial as an anticancer drug in the prevention and treatment of estrogen-dependent mammary tumors. Therefore, this creates interesting possibilities for the clinical applications of melatonin in breast cancer.


Common variable immunodeficiency (CVID) is one of the most common and clinically important primary immune deficiencies. CVID patients have poor humoral immunity, resulting in recurrent infections of the gastrointestinal and upper respiratory tracts, as well as increased incidence of some forms of cancers and autoimmune diseases. The treatment for CVID is IgG replacement, often given as intravenous immunoglobulins (IVIg). IVIg consists of monomeric IgG purified from pooled plasma from healthy donors and is used to treat an increasing number of conditions including autoimmune diseases. In the case of CVID, IVIg has mainly been seen as reconstitution therapy, providing patients with pathogen-specific antibodies. Recent evidence shows that IVIg has diverse effects on the immune system of CVID patients, and one important component is that IVIg alleviates the state of chronic immune activation. In this review, we will discuss causes and consequences of persistent immune activation in CVID, possible underlying mechanisms for how IVIg treatment reduces immune activation, and implications for our understanding of primary as well as acquired immune deficiencies. We will also discuss the similarities between primary and secondary immune deficiencies and the possible implications for our understanding of those diseases.Diagnosis of common variable immunodeficiency (CVID) is based on low levels of IgG and IgA and lack of specific IgG response following vaccination . SeveralCommon variable immunodeficiency patients present defects in several arms of the innate immune system. Natural killer (NK) cells express a wide repertoire of activating and inhibitory receptors and are part of the innate defenses against viral infections and tumors . NK cellE. coli and reduced ROS production after TLR stimulation by PMN from CVID patients. The patients in this study were all under IVIg treatment, and the effects of IVIg on PMN phenotype and function therefore remain undetermined. However, in vitro experiments performed on whole blood from healthy individuals suggest that low doses of IVIg, as used for treatment of CVID, can induce CD11b expression and increase the ROS response (Polymorphonuclear neutrophils (PMNs) are an important component of the innate immune system. In response to pathogens, PMN can rapidly migrate to the site of inflammation and have microbicidal activity by the release of proteolytic enzymes and antimicrobial peptides as well as production of reactive oxygen species (ROS) . In CVIDresponse .bright CD16+ monocytes is elevated and these cells express higher levels of HLA-DR indicating a higher activation level  specialized in capturing, processing, and presenting antigens to initiate immune responses to pathogens. After TLR activation, DCs will mature and increase the expression of co-stimulatory molecules to provide the second signal needed to activate T cells. Bayry et al. showed that into DCs  is defecinto DCs . Howeverloid DCs , and theloid DCs . The frepatients , whereas FcγRIIb . It rema FcγRIIb .+ and a decrease in the CD8+ subset reported in one cohort (in vitro expansion (Because they can rapidly be activated and produce cytokines without previous encounter of their antigen, iNKT cells are sometimes considered to be part of the innate immune system . They ree cohort . The fune cohort . Increasxpansion . The frexpansion , indicatTreatment-naïve CVID patients present low-CD4 T cell counts, in some cases down to numbers that would be considered AIDS-defining in HIV-1 infected patients. Following IVIg initiation, CD4 counts increase in the majority of CVID patients and can reach normal levels in some cases , 31. It Similar to the CD4 T cell compartment, CD8 T cells in treatment-naïve CVID have elevated expression of activation markers Ki67 and co-expression of CD38 and HLA-DR. IgG replacement therapy leads to reduced expression of Ki67, CD38, and HLA-DR on CD8 T cells , indicatRegulatory T cells are key regulators of immune responses and they play a crucial role in limiting unwanted and persistent immune activation. Several studies demonstrated that Tregs are reduced in CVID patients , 36–38 aIn vivo and in vitro studies support the notion that IVIg reduces T cells activation indirectly by acting on APCs rather than on the T cells themselves (Signs of activation in both monocytes and DCs are associated with T cell activation , 22, 40,emselves –45. IVIgCMV infection has been associated with complications in CVID patients , 47 and Some of the immunological perturbations observed in treatment-naïve CVID patients are strikingly similar to those seen in untreated HIV-1 infection. HIV-1 infection leads to chronic immune activation characterized by increased T cell activation and exhaustion, elevated levels of sCD14 and a partial loss of CD4 T cells, DCs, iNKT cells, and Tregs. These features are also found in CVID patients. T cell activation is closely associated with HIV-1 disease progression  but it iex vivo (Intravenous immunoglobulin provides CVID patients with a partial replacement for their defective humoral immunity. However, CVID patients also present abnormalities in cellular immunity in a way similar to what is often seen in other conditions associated with chronic immune activation such as HIV-1 infection. Some of these changes are normalized by IVIg treatment, suggesting that IVIg may also be beneficial in other immune deficiencies characterized by persistent immune activation. IVIg seems to have short-lived effects on the monocyte and Treg populations, whereas the reduction of activation in DCs, iNKT cells, and T cells appear to be sustained. However, the vast majority of immunological studies of CVID have been performed on patients who are already on IVIg replacement, therefore, missing or underestimating some of the abnormalities in CVID that are corrected by IVIg. More studies of treatment-naïve CVID patients are needed to better understand what is driving immune activation in CVID and how IVIg is helping to improve cellular immunity. Furthermore, it will be important to investigate how we can restore the compartments that are not recovering after initiation of IVIg, such as Tregs and iNKT cells. Persistent loss of these cells may help explain why some CVID patients still suffer from severe inflammatory complications, such as interstitial lung disease and autoimmune enteropathy, even when on replacement therapy. Cellular therapy has been safely and successfully used to treat two CVID patients suffering from CMV intestinal disease by injection of autologous specific cytotoxic T cell lines expanded ex vivo . Thus, 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.


Hydroa vacciniforme is a rare idiopathic photodermatosis uncommon in dark skinned patients, which usually begins in childhood before the age of 10 years old and disappear in adolescence, it begins symmetrically in photo exposed areas as a sensation of skin burning in less than 24 hours of sun exposition, and then a vesicular rash develops, become umbilicated and secondarily crusted. Within weeks, the scars heal and leave a residual varioliform scar appearance, the condition generally improves with regular use of high protection sunscreens and resolves during adolescence. In patients who do not respond to conservative treatment, use of systemic agents has been reported like psoralen with exposure to ultraviolet A [PUVA], ultraviolet B TL-01phototherapy, antimalarial agents and immunosuppressive medication. Our patient is a 10-year-old child -without pathological antecedents- who consulted for atrophic depressed scars after a vesicular rash, which begin with a sensation of skin burning a day after intense sun exposure. These scars are covered by hemorrhagic crusts in some areas and appear on exposed areas: face, ear, hands and forearms with erosive lesions of the lower lip. The Child reports that this is the second episode with an interval of 6 months between the 2 episodes.