Analogous to findings in the rat, muscle spindles analyzed were most frequently supplied by two proprioceptive afferents, and in the majority of instances, both were classified as primary endings using established morphological criteria. Secondary endings were most frequently observed when spindle associated afferents totaled three or more. The mean diameter of primary and secondary afferent axons differed significantly, but the distributions overlap more than previously observed in cat and rat studies.Muscle proprioceptive afferents provide feedback critical for successful execution of motor tasks via specialized mechanoreceptors housed within skeletal muscles: muscle spindles, supplied by group Ia and group II afferents, and Golgi tendon organs, supplied by group Ib afferents. The morphology of these proprioceptors and their associated afferents has been studied extensively in the cat soleus, and to a lesser degree, in the rat; however, quantitative analyses of proprioceptive innervation in the mouse soleus are comparatively limited. The present study employed genetically-encoded fluorescent reporting systems to label and analyze muscle spindles, Golgi tendon organs, and the proprioceptive sensory neuron subpopulations supplying them within the intact mouse soleus muscle using high magnification confocal microscopy. Total proprioceptive receptors numbered 11.3 ± 0.4 and 5.2 ± 0.2 for muscle spindles and Golgi tendon organs, respectively, and these receptor counts varied independently ( Continual monitoring of alterations in muscle length, corresponding joint angle changes, and forces produced during muscle contraction are critical for execution of motor tasks. Proprioceptive sensory neurons (PSNs) encode and relay this information to the central nervous system for interpretation and response via spinal circuits and ascending pathways into the brain ,2. AxonsMS and GTOs are both encapsulated, stretch-activated sensory receptors found within skeletal muscles. As a consequence of their differing intramuscular location and architecture, however, the PSNs that innervate each receptor respond to distinct physical stimuli. For example, MS are located in the belly of the muscle, positioned in parallel with extrafusal muscle fibers causing them to respond to muscle stretch . ConversMS are structurally more complex and are generally comprised of three types of intrafusal muscle fibers  surrounded by a capsule structure . The MS As a result of their distinct endings on the muscle spindle, group II afferents encode stretch-evoked stimuli differently than Ia spindle afferents. For example, Ia afferents display a supralinear action potential firing response during passive muscle stretch, together with a characteristic pause in firing when the muscle is shortened . Firing To date, knowledge of the anatomy and physiology of MS and GTO has come primarily from work in the cat, and secondarily from studies in rat ,8,10,16.Rosa26tdTom/+ termed here R26-tdT; JAX Stock 007908) was crossed with two Cre-driver lines to label either all peripheral sensory neurons (Advillin-Cre (Adv-Cre); ) PSN axons in total. Our analysis further revealed 2 to 7  PSN axons within the thin branch. In all cases, Ib axons were present in the thin branch of the soleus nerve. Spindle afferents, however, were not always present in the thin branch, ranging from 0 to 5 axons .In every case, confocal imaging confirmed individual GTOs were innervated by a single PSN axon, namely a Ib afferent  = 0.498; rs(7) = -0.536; Possessing complete knowledge of the number of MS and GTOs, along with the number of PSN axons innervating these structures in multiple soleus muscles, we investigated the relationship between PSN axons and associated endings in muscles with varying numbers of MS and GTOs. Larger numbers of MS and GTOs were associated with increased numbers of proprioceptive afferents innervating the muscle, although the correlation was weak  was innervation by a single primary afferent. The frequency of the other two main categories, innervation by only two primary afferents  or by a primary with two secondary afferents , were similar  from age P19-20 soleus muscles were sorted according to their number of total associated proprioceptive afferents . Varying configurations of primary (P) and secondary (S) endings identified from morphological criteria were observed when MS had two or more associated afferents. In cases where two afferents supplied a MS, two primary endings (PP) were most frequently observed (14/22 MS). When three afferents supplied a MS, however, the configuration shifted to favor multiple secondary endings .Adv-Cre; R26-tdT mice. We first analyzed the distribution of diameters for multimodality and found the data to be statistically normal. Next we compared the axon diameters of afferents that must be either group Ia (afferents supplying the 30.0% of MS receiving a single afferent axon) or group Ib afferents innervating GTOs. The variances of the Ib afferent and Ia afferent diameter samples were equal. Therefore equal variance, two-tailed t-tests were performed, indicating that the average diameter of Ib afferents  was not statistically different from the average diameter of Ia afferents supplying single-axon spindles .In addition to characteristic differences in contacts with intrafusal fibers, Ia afferents in cats also have a larger average axonal diameter than group II afferents . We nextTo be conservative in the initial assessment of group II afferents, only the smallest axon taken from the subset of MS innervated by three axons was analyzed (n = 17). After unblinding the reviewers, 15 of these 17 axons were shown to have been classified as secondary endings based on morphological criteria. The two axons whose morphological classification and diameter-based prediction differed were removed from the group of presumed group II axons. The average diameter of the remaining group II axons was significantly smaller than the population of presumed primaries . A logistic regression was then applied to all spindle afferents to estimate the likelihood of correctly classifying a single axon as either a Ia or group II axon, based on diameter alone. MS afferents were first classified as primary or secondary endings based on morphological criteria and then these identifications were compared to those predicted by the logistic regression model. The model demonstrated that afferents could be correctly classified by diameter alone 67% of the time (p = 0.0011). To determine diameter cut-offs that would most accurately predict a spindle afferent’s subtype, threshold values were varied. The model had the strongest specificity and sensitivity, 74% and 72% respectively, when the thresholds were set to probabilities greater than 65% and less than 30%. These threshold values corresponded with the diameter measurements of 2.91 μm and 2.46 μm see . Thus, aThe logistic regression was conducted because Hartigan’s dip test determined that the combined population of all spindle afferents was statistically normal even though previous literature has reported that axon diameters follow bimodal distributions in the cat . ConversBoth morphological and axon diameter approaches contribute to our understanding of the afferent types innervating individual MS. Applying this information to the total complement of MS (11.3) and GTOs (5.2) found in the soleus, it was possible to estimate the total number of Ia, Ib, group II axons that innervate the soleus. Given GTOs were always observed to be innervated by a single Ib afferent, approximately 5 Ib afferents project to the soleus muscle. From the subset of MS analyzed in In this study we used a transgenic reporter system to analyze the sensory innervation of MS and GTOs using an intact mouse soleus muscle preparation. Robust fluorescence in PSN axons permitted visualization of afferent pathways and terminations within a muscle with a minimum of specimen preparation and no tissue sectioning, representing a substantial gain in efficiency compared with serial section or teased preparation techniques. This whole-mount muscle approach enabled quantification of proprioceptive receptors and their respective afferents as well as comparison of spindle sensory ending classifications obtained via morphological identification and intramuscular axonal diameter measurement.PV-Cre or Adv-Cre driver lines, the R26-tdTomato reporter line generated strong fluorescence in sensory axons that required no amplification following brief fixation. Use of the PV-Cre was desirable because among muscle sensory afferents, parvalbumin expression is limited to only proprioceptive axons [Adv-Cre eliminated expression in muscle fibers, and revealed a dense network of sensory axons and endings throughout the muscle. In addition to PSN axons, smaller caliber axons (likely group III and IV) were widely distributed in the muscle [When controlled by either ve axons ,23. Durive axons  leads toe muscle . These ae muscle . Such a MS in the mouse soleus have been quantified in previous reports, usually as part of a larger research question with fewer replicates than the present study. Our reported average of 11.3 ± 0.4 MS occupies the middle of the range of numbers from 10 to 12 MS reported elsewhere ,33–38. GNT-3 heterozygous animals [Analysis of MS and GTOs using an intact muscle allowed us to readily quantify endings from a relatively large sample of muscles, revealing a wide range of natural variability in both the number of MS, GTOs, and the PSN axons supplying these structures. Could the natural range of proprioceptive endings, two-fold for MS (8 to 16) and GTOs (3 to 7) influence the proprioceptive sensitivity of individual animals? Total loss of muscle spindles has an obvious impact on locomotor behavior, but intermediate reductions in spindle number have less obvious effects. Muscle spindles do not form in mice lacking neurotrophin-3 (NT-3), a trophic factor necessary during embryogenesis for PSN survival, and approximately half the normal complement of muscle spindles are found in mice heterozygous for the mutation . Neverth animals . This su animals ,41. Neve animals .The uncorrelated variability in MS and GTO numbers in individual muscles suggests no consistent ratio of MS and GTOs is maintained in the soleus. While the mechanisms involved in MS and GTO induction remain unclear, alterations in gene expression can favor development of one receptor type at the expense of the other. In mice lacking Er81, an ETS-family transcription factor expressed by all PSNs and also induced in intrafusal fibers during development, the number of MS in the soleus increased while the number of GTOs decreased ,36. DualWe found muscle spindle primary (Ia) and secondary (II) afferent populations of mature mice are significantly different in terms of intramuscular diameter, but the distributions share substantial territory. Overlap in the diameters of group Ia and II axons is also observed in the cat, with up to 75% of soleus group II axons being found within the range of Ia afferent axon diameters . The degTo date, the most detailed anatomical analysis of the afferents supplying MS in rodents has come from rats, and this data suggested spindles in the soleus muscle frequently receive multiple primary afferent endings (~65% of spindles), a finding in agreement with our results, but which contrasts with the predominant single primary afferent pattern observed in the classic feline animal model . The preIn summary, genetic reporter lines were used in this study to comprehensively analyze the proprioceptive components of the mouse soleus muscle. These mouse lines enabled us to examine a whole soleus muscle, accelerating the process of quantifying MS and GTO counts. Receptor counts demonstrated that there are approximately twice as many MS as GTOs on average, and that the number of each receptor type varies independently from the other in the soleus. Morphological analysis of PSN axons revealed that most muscle spindles are innervated by two or three axons, with variable numbers of primary and secondary endings. Overall, morphological classifications of spindle afferent subtypes matched classifications by axon diameter. However, due to the substantial degree of overlap in the diameters of Ia and II axons, over a third of afferents could not accurately be classified by their diameters. Combined with findings in the rat, the overlapping ranges of Ia and II axon diameters in this study suggest that in the rodent, conduction velocity and axon diameter only distinguish afferent subtypes at the farthest extremes of data distributions, and therefore reinforce the need for careful morphological characterization or more robust electrophysiological techniques to accurately classify primary and secondary endings . Taken t


Mycobacterium is a genus of aerobic and acid-fast bacteria, which include several pathogenic organisms that cause serious diseases in mammals. Previous studies have associated the immune response against mycobacteria with multiple sclerosis (MS), a chronic demyelinating disease of the central nervous system with unknown etiology. The role of mycobacteria in the pathological process has been controversial and often conflicting. We provide a detailed review of the mycobacteria that have been linked to MS over the last three decades, with a focus on Mycobacterium bovis bacille Calmette–Guérin vaccine for human and oral exposure to Mycobacterium avium subsp. paratuberculosis. We will also discuss the exposure and genetic susceptibility to mycobacterial infection, the protective role of vaccination, as well as the possible mechanisms involved in initiating or worsening MS symptoms, with particular emphasis on the molecular mimicry between mycobacterial and human proteins. Finally, we will introduce topics such as heat shock proteins and recognition by innate immunity, and toll-like receptor signaling-mediated responses to Mycobacterium exposure. Multiple sclerosis (MS) is a chronic, inflammatory demyelinating disorder of the central nervous system (CNS) characterized clinically by a wide variety of neurological symptoms .Even though rigorous research has been performed in the MS field, its etiology as well the exact pathogenic mechanisms remains poorly understood. Nevertheless, it is believed to be a multifactorial disease caused by autoimmune processes . While tEpstein–Barr virus (EBV)  , 3, endooversial .Innate immunity plays an important role in the initiation and progression of MS, as well as in host defense against mycobacteria infection and their immunogenic components Figure  4)..4).Mycobacterium and pathogen-associated molecular patterns that activate innate immune responses through TLRs such as TLR2, TLR4, and TLR9  priming can promote protection against EAE .Mycobacterium tuberculosis and Mycobacterium leprae in MS, compared to that against specimens isolated from patients with other neurological diseases or healthy control subjects  HSP70 antibodies in Sardinian patients with MS  , and a lMycobacterium avium subsp. paratuberculosis, an intracellular pathogen belonging to the Mycobacterium avium complex, is the causative agent of paratuberculosis or Johne’s disease  of 15.5% of the 436 patients with MS, and in 2.3% of the 264 healthy controls by PCR amplification of the highly specific equences .Further, the possible involvement of MAP in the etiology and pathogenesis of MS was supported by the discovery of several MAP proteins and antigenic peptides capable of inducing a stronger humoral and cell-mediated response in the PBMCs of patients with MS than in those of control subjects , 22.The humoral response against several homologous epitopes of EBV, MAP, and human proteins has also been explored –27. Anti295-303, an immunodominant B-cell epitope belonging to a MAP-specific protein, which is located within the high homology region with the T-cell receptor gamma-chain protein -specific IFN-γ+, CD4+, and IL-17+CD4+ T cells following EAE induction  for a 6-month period before starting the immunomodulation therapy . CIS des35-55 epitope and BCG-derived homologous peptides showed the absence of antibody-recognition against BCG peptides in 40 Sardinian patients with MS (Few studies have investigated the humoral response elicited by BCG antigens. A seroprevalence study based on the antibody detection against the encephalitogenic MOG with MS .Overall, our findings demonstrated a relationship between mycobacteria and MS. Cell-mediated immunity plays a critical role in protection against mycobacteria as they are intracellular pathogens, and even if the exact mechanisms underlying the effects of BCG on neuroinflammation are unclear, a protective role of BCG vaccination on MS progression is generally accepted. Further research is needed for potential therapeutic use of the BCG vaccine in patients at risk of developing MS.Mycobacterium avium subsp. paratuberculosis seems to have a causal role in the MS pathology according to the molecular mimicry theory in some genetically predisposed individuals, and clinical trials of antimycobacterial therapy targeting MAP are currently under way (nder way . The rolDC performed the experiments related to several articles included in this mini review and wrote the manuscript; KY wrote the manuscript and supported in the critical reading; NH wrote the manuscript and supported in the critical reading.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.


We report a very unique case of an esophageal metastasis from a pancreatic ductal adenocarcinoma (PDAC) primary.After obtaining consent from the patient, all relevant records of the case were obtained and retrospectively reviewed.At presentation, the patient was diagnosed with synchronous pancreatic and esophageal cancer. He received six months of neoadjuvant therapy including FOLFIRINOX  and stereotactic body radiation therapy (SBRT) to the pancreatic tumor followed by a combined pancreaticoduodenectomy and Ivor Lewis esophagectomy. Review of the final esophageal specimen revealed normal overlying squamous mucosa with an underlying focus of metastatic PDAC. The patient remains alive with no evidence of disease 17 months from surgery and 25 months from diagnosis.Differentiating an esophageal metastasis from a PDAC primary versus a synchronous esophageal carcinoma is very difficult despite state-of-the-art diagnostic techniques performed at a high-volume tertiary cancer center. Extensive evaluation and continued follow-up of PDAC patients presenting with a synchronous esophageal lesion in a multidisciplinary setting may help ensure efficient and accurate management. In our case, neoadjuvant FOLFIRINOX and SBRT to the primary PDAC tumor followed by surgery has been an effective approach for this patient. Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of death by a solid malignancy in the United States, with a 5-year overall survival rate of 8%.  PDAC is PDAC most commonly metastasizes to lymph nodes, the liver, lung, and peritoneal cavity, while rare locations that have been reported include bone, brain, myocardium, and the umbilicus. , 4 At thnd case of a metastasis to the esophagus arising from a PDAC primary reported in the modern era (since the 1980s). [Not only is a PDAC metastasis to the esophagus extremely rare, but it is also difficult to distinguish an esophageal primary from a metastasis to the esophagus by radiographic imaging or endoscopy. To our knowledge, we report the 2 1980s). -13A 72-year-old non-smoking male presented with a 6-month history of weight loss (9 kg) followed by obstructive jaundice characterized by a 2-month history of acholic stools and dark urine. Past medical history was significant for hypertension and dyslipidemia and an extensive family history of cancer was significant for pancreas, liver, breast, gynecologic, and colon malignancies in 5 siblings and his father. Initial evaluation was conducted by his primary care provider and included laboratory studies and imaging. Computed tomography (CT) scan of the abdomen and pelvis revealed a 2.5 x 1.7 cm mass in the pancreatic head, abutment of the superior mesenteric artery (SMA) and vein (SMV), and marked biliary and pancreatic ductal dilatation consistent with PDAC. Liver function tests (LFTs) were elevated, with an alkaline phosphatase of 515 IU/L, aspartate aminotransferase of 198 IU/L, and total bilirubin of 10.3 mg/dL. Carbohydrate antigen 19-9 (CA 19-9) at this time was 395 U/mL. Upon further workup by a gastroenterologist, endoscopic ultrasound (EUS) with fine needle aspiration (FNA) revealed adenocarcinoma of the pancreatic head in addition to an incidental 2.0 cm distal esophageal exophytic lesion that returned positive for adenocarcinoma. The relationship of these two carcinomas was uncertain. Endoscopic retrograde cholangiopancreatography (ERCP) was also performed for metallic biliary stent placement to relieve high-grade biliary obstruction related to the pancreatic mass.Further imaging with 18-fluorodeoxyglucose positron emission tomography (FDG-PET)/CT demonstrated a large hypodense mass in the head of the pancreas with moderate FDG activity consistent with the patient’s known PDAC in addition to multiple enlarged peripancreatic, aortocaval, and porta hepatic lymph nodes as well as a focal area of mild metabolic activity in the distal esophagus just above the gastroesophageal junction with multiple paraesophageal lymph nodes. At an outside institution, the patient was diagnosed with localized PDAC that was thought to be unresectable along with a separate esophageal adenocarcinoma primary. Therefore, his local oncologist recommended chemotherapy with FOLFIRNOX  and referred the patient to our Pancreatic Multidisciplinary Clinic (PMDC) for additional recommendations. nd and 3rd portions of the duodenum and demonstrating proximal main pancreatic duct dilation. Vessel involvement included encasement of the SMV/portal vein (PV) confluence and 180° abutment of the SMA, thereby conferring a diagnosis of borderline resectable PDAC  or stereotactic body radiation therapy (SBRT) with re-evaluation for potential surgical resection. FOLIFIRINOX was the recommended chemotherapy such that the platinum agent would have activity in both primary pancreatic and esophageal tumors. Depending on the expertise of the thoracic oncologists and tumor response to chemotherapy, standard CRT would be warranted in order to encompass both the esophagus and pancreas in the same field; however, if the esophageal lesion would not require neoadjuvant radiation, SBRT to the pancreas lesion would be preferred. In order to address the suspected esophageal lesion, our thoracic colleagues were consulted and the patient was referred for formal evaluation by a thoracic surgeon.After endoscopy and thoracic surgical consultation, the esophageal lesion was thought to be a synchronous esophageal primary cancer  and the treatment recommendation consisted of neoadjuvant FOLFIRINOX followed by SBRT and evaluation for surgery. It was understood that treatment of the PDAC was of primary significance, with the possibility of delivering definitive CRT to the esophagus later in the treatment course.The following week, FOLFIRINOX was initiated locally and continued for 3 months . Following 6 doses of FOLFIRINOX, the patient presented back to our PMDC for re-evaluation. The patient continued to work 12-hour days throughout therapy, with his only complaint being minor fatigue. His CA 19-9 had decreased to 71.9 U/mL at this time (4 months from diagnosis), with CT demonstrating the pancreatic mass and regional lymphadenopathy to be slightly less bulky, improvement of SMA/SMV involvement Figure , and impThe patient then underwent SBRT to the pancreatic tumor to a total cumulative dose of 30.5 Gy in 5 fractions. Image guidance was performed using 3 gold fiducial markers endoscopically placed around the lesion and active breathing control (ABC) was used to minimize movement of the tumor during respiration. Images of the pancreatic and esophageal lesions at the time of endoscopy can be visualized in Figure Of note, an esophagogastroduodenoscopy (EGD) was performed at the time of endoscopic fiducial placement to re-biopsy the esophageal lesion. The morphology was most consistent with a carcinoma that spread from the pancreaticobiliary system and immunolabeling for SMAD4 demonstrated retention of labeling, which neither confirmed nor refuted an interpretation of spread from a pancreaticobiliary lesion. The patient also experienced a few episodes of hematochezia during chemotherapy. A colonoscopy was performed and reported as negative, with the bleeding resolving spontaneously.Eight months after initial diagnosis and after six months of neoadjuvant therapy, the patient underwent a pylorus-preserving pancreaticoduodenectomy and Ivor Lewis esophagectomy with jejunostomy feeding tube (J-tube) placement. During the operation, the right gastric artery was preserved and the blood supply to the stomach was confirmed both visually and with an intraoperative Doppler ultrasound. The pancreatic specimen revealed numerous microscopic foci of adenocarcinoma with vacuolated cytoplasm and hyperchromatic nuclei scattered within a 5 cm fibrotic tumor bed Figure , otherwiin situ carcinoma. Focally active chronic gastritis and chronic cholecystitis were noted. The normal overlying squamous mucosa as shown in Figure The esophageal specimen revealed a microscopic focus (3 mm) of infiltrating adenocarcinoma involving submucosa of the distal esophagus Figure  along wiThere were no major surgical complications although gram-positive cocci in chains were found in the wound and treated with antibiotics and negative-pressure wound therapy was performed for drainage at the incision site. One month from surgery, the patient was doing well with no major complaints other than intermittent abdominal pain. His J-tube was removed at the time of two month follow-up and adjuvant chemotherapy was recommended; however, numerous appointments were missed and the patient refused adjuvant therapy. Unintentional weight loss secondary to pain during eating and acid reflux led to a nutrition consultation and prescription of a proton pump inhibitor and pancreatic enzyme supplementation. The patient occasionally complains of abdominal pain and nausea but is doing well otherwise and remains with no evidence of disease 17 months from surgery and 25 months from diagnosis, with surveillance laboratory studies and imaging occurring every 3 months.th reported case of an isolated esophageal metastasis from PDAC and the 2nd report on an aggressive approach of a combined pancreaticoduodenectomy and esophagectomy. [This case report summarizes a unique case of a patient with PDAC with an isolated metastasis to the esophagus who underwent successful resection of a pancreatic head and distal esophageal lesion after neoadjuvant therapy. To our knowledge, this is the 12gectomy. -13, 15The patient’s pancreatic primary showed moderate treatment effect among a background of extensive fibrosis, whereas the esophageal lesion showed a more classic appearance of an untreated adenocarcinoma with numerous dilated glands with an infiltrative growth pattern, nuclear atypia, and scattered mitotic figures. While there was focal Barrett’s mucosa elsewhere in the esophagus, it did not show significant dysplasia. Most significantly, the adenocarcinoma in this case had overlying squamous mucosa with no significant histopathologic change. While SMAD4 expression was intact in the esophageal lesion, roughly half of PDACs retain SMAD4 expression, and this result does not exclude spread from a PDAC primary. , 17versus a synchronous esophageal carcinoma. Despite numerous methods of imaging and procedures including CT, PET/CT, EUS, EGD, and immunohistochemistry as well as multidisciplinary review among radiology, pathology, gastroenterology, surgery, medical oncology, and radiation oncology at a high-volume tertiary center, the final diagnosis of metastatic PDAC to the esophagus was not reached until a substantial amount of tissue was reviewed after partial esophagectomy. FDG-PET is commonly used in combination with CT and/or EUS to identify occult metastases in pancreatic and esophageal adenocarcinomas. However, the sensitivity and specificity of detection of metastases range from 50-90% and, as observed in this case report, may not lead to conclusive evidence. [This case report elucidates the difficulty of differentiating an esophageal metastasis from PDAC primary vidence. -21In general, local therapy is not traditionally recommended for metastatic disease, PDAC or otherwise; however, oncologists are becomingly increasingly aggressive by offering radiation therapy and/or surgical resection in this patient population, particularly in the setting of limited oligometastatic disease. , 22-27 Ivs. 6.8 months), progression-free survival (6.4 months vs. 3.3 months), and objective response (31.6% vs. 9.4%). Since then, FOLFIRINOX has been studied in other settings of PDAC as well as other gastrointestinal cancers, with promising response rates in patients who are able to tolerate the regimen. [Neoadjuvant FOLFIRINOX was administered in this patient as a method to provide aggressive systemic therapy and include a platinum agent that may warrant a treatment response in both the PDAC and esophageal lesion. In 2011, Conroy and colleagues published the results of a randomized clinical trial comparing FOLFIRINOX and gemcitabine monotherapy.  FOLFIRINregimen. -35An esophageal metastasis from a pancreatic primary may be more common than traditional belief and patients with a suspicious esophageal lesion should undergo comprehensive evaluation and close follow-up in order to guide management. Although there are limited data to suggest an optimal approach to these cases, neoadjuvant FOLFIRNOX followed by SBRT and surgery has resulted in favorable disease control over two years from diagnosis despite no adjuvant therapy. However, we only recommend aggressive surgery of both lesions if there is no clear evidence of metastatic disease at other locations after an extended period of time ( > 6 months). As future technologies involving novel analytic techniques such as next-generation sequencing, , 37 circAfter obtaining consent from the patient, all relevant records of the case were retrospectively reviewed. The pathologic specimens were reviewed with response to neoadjuvant therapy graded using the criteria described previously by the College of American Pathologists.


HOX and PBX gene families encode transcription factors that have key roles in establishing the identity of cells and tissues in early development. Over the last 20 years it has become apparent that they are also dysregulated in a wide range of solid and haematological malignancies and have a predominantly pro-oncogenic function. A key mode of transcriptional regulation by HOX and PBX proteins is through their interaction as a heterodimer or larger complex that enhances their binding affinity and specificity for DNA, and there is growing evidence that this interaction is a potential therapeutic target in malignancies that include prostate, breast, renal, ovarian and lung cancer, melanoma, myeloma, and acute myeloid leukaemia. This review summarizes the roles of HOX and PBX genes in cancer and assesses the therapeutic potential of HOX/PBX dimer inhibition, including the availability of biomarkers for its application in precision medicine.The  HOX genes are a family of homeodomain-containing transcription factors that were originally identified due to their pivotal roles in early development [HOX-mutant animals, most notably Drosophila. One of the best known examples of this is the ectopic expression of the Antennapedia (Antp) gene in cells that would normally give rise to antennae; a near perfect pair of legs replaces the antennae of adult flies that develop from these embryos [HOX genes can determine the identity of cells and tissues, and hence also help regulate the proliferation, differentiation, and survival of these cells. The identification of additional mutants revealed a family of 8 HOX genes in the fly, which are expressed in an anterior to posterior pattern along the main embryonic axis and specify the identity of different embryonic structures [The elopment . These a embryos . These oructures .HOX gene organization is their existence in clusters within a single chromosome, and 2 further rounds of duplication in the course of vertebrate evolution have given rise to 4 chromosomal clusters in mammals, referred to as A, B, C, and D. The cluster names are also used to denote specific genes in conjunction with the position of the gene in the cluster, thus for example HOXD1 is the most 3′ member of the D cluster [HOX gene expression, as do a number of microRNAs encoded within each cluster [HOX regulation, which is in part enabled by enhancer sharing, is temporal and spatial collinearity during development, whereby each HOX gene in a cluster is expressed earlier in development and with a more anterior border of expression than its 5′ neighbour [An unusual feature of HOX genes that play key roles in patterning both the main embryonic anterior to posterior axis at a very early stage of development and embryonic structures that develop later, for example the limbs and many of the organs [HOX genes is reflected in a high level of functional redundancy during development [HOX genes playing specific roles in the embryo, for example during limb development where members of the HOXD cluster define specific structures [In total mammals have 39 elopment , althougructures .HOX genes were originally characterized as developmental genes they also play a number of important roles in the adult, and indeed their original embryonic expression patterns are sometimes maintained, at least to a limited extent [HOX gene function in the adult include the maintenance of hematopoietic stem cells (HSCs) [HOX genes become highly dysregulated (and often over expressed) in a wide range of both solid and haematological cancers.Although d extent . The moss (HSCs) , the spes (HSCs) , and regs (HSCs) . In addiS100B gene to promote its transcription in neuroblastoma cell lines [ITGB3 gene (encoding Integrin β3) during angiogenesis [TWIST1 enhancer, thereby repressing TWIST1 transcription [Although HOX proteins can bind to DNA through their homeodomain, this binding is relatively non-specific as it generally involves only a 4 base pair recognition sequence. Greater specificity is conferred by the binding of cofactors such Pre-B-cell Leukemia Homeobox (PBX) and Myeloid Ecotropic Viral Integration Site 1 Homolog (MEIS) proteins . The latll lines , whilst ogenesis . Converscription .In addition to determining target gene specificity and the mode of transcriptional regulation, HOX cofactors also play a role in post-translational regulation through facilitating the entry of HOX proteins into the nucleus (considered in more detail below).HOX genes in cancer first became apparent from their frequent inclusion in chimeric, oncogenic gene fusions that drive the formation of haematological malignancies [HOX genes are profoundly dysregulated in a wide range of both solid and haematological malignancies, most frequently showing very high levels of over expression. There is now a vast amount of data available on HOX gene expression in different malignancies, and it is not within the scope of this review to detail this. The reader is instead referred to a number of recent specialist reviews that examine the expression and possible function of HOX genes in specific cancer types such as melanoma [HOX dysregulation in cancer, relatively few HOX genes have been shown to act as oncogenes in the strictest sense, i.e. that their forced expression in phenotypically normal cells is sufficient to cause a switch to a malignant phenotype, although an important example is HOXA9, which can immortalize normal bone marrow cells in mice when overexpressed [HOX genes have a general supportive role in malignancy, both at the cellular level (for example in promoting proliferation and blocking apoptosis) [HOX expression being associated with a poor clinical outcome and prognosis (considered in more detail below).A potential role for gnancies . It has melanoma , and heamelanoma , prostatmelanoma , breast melanoma , ovarianmelanoma , and panmelanoma . Despitexpressed . Currentoptosis) , and at optosis) , drive moptosis) , and facoptosis) , 28, 29 optosis) . These pHOX genes also act as tumour suppressors. Examples include HOXA4, the expression of which can block the spread of ovarian cancer cells [HOXC8 which is inversely related to progression in ovarian cancer [In addition to a pro-oncogenic role, a number of er cells , HOXA5 ter cells  and promer cells , and HOXn cancer . Intrigun cancer , 34, or n cancer .HOX genes in cancer make the products of these genes potential biomarkers for both diagnosis and prognosis, and for precision medicine applications such as predicting treatment response , a gene that is very closely related to the HOX genes but which is not located within the 4 main chromosomal clusters described above [HOXC6 RNA has been shown to be a potential diagnostic marker for prostate cancer as part of a multi gene panel [HOX gene expression at the RNA level has also been shown to have prognostic significance in acute myeloid leukaemia (AML) [HOX gene expression apparently exist within different types of cancer, indicating that they could be used to distinguish between cancer types when the primary tumour type is unknown, for example in circulating tumour cells (CTCs) [The differential expression of ni, 2016 ). One ofed above . EN2 is ed above , 39. In ne panel . HOX genia (AML) , and in ia (AML) , breast ia (AML) , ovariania (AML) , oral sqia (AML) , thyroidia (AML) , clear cia (AML) , gastricia (AML) , and blaia (AML) . Discrees (CTCs) .PBX genes are homologues of the Drosophila extradenticle gene (Exd) and 4 are encoded in the human genome (PBX1-4). Like the HOX genes they encode homeodomain-containing transcription factors, but do not exist in chromosomal clusters. In addition to the homeodomain, PBX proteins contain other highly conserved regions, one of which is required for binding to a number of closely related transcription factors, MEIS and PREP [and PREP . PBX proand PREP . The extand PREP . Unusualand PREP .PBX proteins form strong complexes with HOX1-11 proteins in the presence of a HOX/PBX DNA binding consensus –55. X-raHOX genes in the context of cancer, the PBX gene family is known to have a number of oncogenic functions. Best characterised of these is as a chimeric fusion partner in various leukaemias and lymphomas, for example PBX1 and E2A in pre-B cell acute lymphoblastic leukaemia [PBX3 together with MEIS1 is necessary to drive leukemogenesis efficiently in mouse models; PBX3 protein is required both to stabilize MEIS1 and induce transcription of the MEIS1 gene [PBX3, HOXA7, HOXA9, and HOXA11 has been shown to be an independent predictor of poor survival in patients with cytogenetically abnormal AML (CA-AML), and that PBX3 (but not PBX1 or PBX2) is frequently co-expressed with HOXA9 in various subtypes of CA-AML, particularly MLL-rearranged AML, and may thus be a potential pathologic cofactor of HOXA9. This is further supported by the finding that knock-down of PBX3 prevents it from exerting a synergistic effect with HOXA9 in promoting leukemogenesis [Although less studied than the eukaemia . It has IS1 gene . Correspogenesis .PBX genes are also over expressed in a number of solid tumours. These include colorectal cancer (CRC), in which PBX3 expression was shown to be correlated with invasive potential in vitro, and significantly associated with lymph node invasion, distant metastasis, advanced TNM stage and poor overall survival of patients. Furthermore, the forced expression of PBX3 in cells with a low metastatic potential was shown to promote migration and invasion, at least in part through the upregulation of phosphorylated extracellular signal-regulated kinase (ERK)1/2 [PBX3 has also been shown to be upregulated in gastric cancer cells, and that this was associated with greater invasion depth, and advanced clinical stage and tumour grade. The overexpression of PBX3 in gastric cancer cell lines with low endogenous PBX3 expression accelerated cell proliferation and increased colony formation and cell-invading ability [PBX3 expression have been reported in prostate tumours and this was found to have prognostic significance in this cancer. PBX3 expression is promoted by androgen signalling through a pathway that is negatively regulated in part by the Lethal-7 family of microRNAs (let-7), which are in turn downstream targets of the Myc protoncogene [PBX3 as the most oncogenic PBX gene, a recent study demonstrated that increased PBX1 expression in ovarian cancer was associated with shorter post-chemotherapy survival and increased resistance to platinum-based drugs through the maintenance of a stem cell-like phenotype [The (ERK)1/2 . PBX3 ha ability . Similaroncogene . Althoughenotype .The key roles that HOX and PBX proteins play in cancer indicate that they are potential therapeutic targets. However, the high level of functional redundancy amongst HOX proteins and the general difficulty in producing effective small molecule inhibitors against transcription factors have proved significant barriers to this approach. As an alternative, it was proposed that the interaction between HOX and PBX proteins could be targeted, as this is mediated by a highly conserved hexapeptide sequence in HOX proteins and a hydrophobic binding pocket within PBX. Although a small molecule inhibitor of this interaction has previously been described, its Kd was in the micromolar range and it does not seem to have been adopted for experimental or clinical use . To datecFos expression, as preventing this using gene knock down strategies can achieve a partial rescue from HXR9-mediated cell killing [cFos has been shown to cause apoptosis in a number of different cancers [cFos tumour expression are associated with longer survival in ovarian cancer patients [cFos is an early response gene to many types of cellular stress, although there is now evidence to suggest that HOX/PBX dimers repress cFos expression both directly, through HOX/PBX binding sites in its promoter, and indirectly through HOX/PBX mediated regulation of a cFos-targeting microRNA [The mechanism by which HOX/PBX inhibition causes cell death remains to be fully elucidated Figure . In most killing . The upr cancers –83, in a cancers , 79–82. patients , and, inpatients . cFos ismicroRNA .HOX genes HOXB1 through to HOXB9 [HOX genes with reported pro-oncogenic functions to those with tumour-suppressor functions [As with other types of targeted therapy, companion diagnostics are becoming increasingly important for patient stratification in clinical trials as well as personalized therapies. The identification of markers that can predict which tumours will be sensitive to HOX/PBX inhibition is an important preclinical goal, although to date there is relatively little evidence for a single or even multiplex marker that can fill this role outside of limited, cancer-specific settings. In a study on breast cancer, the sensitivity of breast cancer cell lines to killing by HXR9 was shown to be strongly associated with the combined expression of to HOXB9 . Howeverunctions . Neitherunctions  and mesounctions  a subgrounctions .The lack of a robust and generalized marker of tumour sensitivity to HOX/PBX inhibition might be related to the difficulty in quantifying these dimers in cells and tissues, as opposed to measuring HOX gene expression at the RNA or protein level, which is at best an indirect measure of this. Work is continuing to develop a quantitative assay for HOX/PBX dimers, and it is hoped that this will in turn lead to a more accurate test for tumour sensitivity to HOX/PBX inhibition.cFos, as described above, together with a number of other targets that might act as a readout of the efficacy of inhibitors such as HXR9 [Surrogate markers of tumour response to treatment, which allow an earlier assessment of the efficacy of treatment than conventional clinical endpoints  are of growing importance both in clinical trials and in precision medicine. The regulatory function of HOX/PBX dimers means that there are a number of immediate transcriptional targets that change in response to HOX/PBX inhibition. These include  as HXR9 . The bes as HXR9 , and dec as HXR9 . ATF3 al as HXR9 –90.in vitro and in vivo data supporting the therapeutic potential of inhibiting HOX/PBX dimer formation in cancer. To date, however, the only effective inhibitors of HOX/PBX binding are the HXR9 peptide and its derivatives. Peptide-based therapeutics are already used in cancer therapy though [There is now considerable y though , includiy though , and indy though . DevelopSince the publication of the initial study in 2007 it has become increasingly apparent that the HOX/PBX dimer is a potential therapeutic target in both solid and haematological malignancies. Clinical exploitation of this target is aided by the co-development of markers that predict the sensitivity of tumours to HOX/PBX inhibitors, and which allow the initial response to treatment to be monitored. The outcome of clinical trials for peptide and small molecule inhibitors of HOX/PBX are eagerly awaited.


Although the general role of these calcium channels in modulating synaptic plasticity and memory has been explored, their role in olfactory learning is not well understood. In this review article we first discuss the role of LTCCs in olfactory learning especially focusing on early odor preference learning in neonate rodents, presenting evidence that while NMDARs initiate stimulus-specific learning, LTCCs promote protein-synthesis dependent long-term memory (LTM). Norepinephrine (NE) release from the locus coeruleus (LC) is essential for early olfactory learning, thus noradrenergic modulation of LTCC function and its implication in olfactory learning is discussed here. We then address the differential roles of LTCCs in adult learning and learning in aged animals.L type calcium channels (LTCCs) are prevalent in different systems and hold immense importance for maintaining/performing selective functions. In the nervous system, CaV CaV1.3 has a lower activation voltage compared to CaV1.2 , characterized by their long-lasting activity and sensitivity to dihydropyridine, are members of the high voltage gated calcium channel family. Like other calcium channels of this type, they are composed of multiple subunits including α1, the pore-forming subunit containing the voltage sensor, which dictates the nomenclature of the channel subtype. Between CaVIn this review article, we present evidence of the functional roles of LTCCs in synaptic plasticity and learning, focusing on olfactory learning as a model system. The role of norepinephrine (NE) via β-adrenoceptors (βARs) in modulating LTCCs and early odor preference learning are detailed. Finally, the differential roles of LTCCs in learning during development and aging are discussed.LTCC dependent calcium entry induces varied downstream molecular cascades, the functions of which are vital to understanding the importance of the channels. The C-terminal of the LTCC contains micro-clusters of AKAP, PKA, calcineurin and calmodulin in its vicinity. Following channel opening and calcium entry, unique combinations of these enzymes can be activated  of synaptic activity. LTP induction in perforant- dentate gyrus synapses is inhibited by chronic administration of an LTCC antagonist  and short-term memory (STM). Blocking PKA or protein synthesis produces a STM for 3 h which is not sustainable at 24 h  are critical for this early olfactory preference learning to occur  current and its slow component has LTCC dependance, especially on CaVAlthough it seems clear that NE and LTCCs are important players in olfactory learning and memory, several questions remain unanswered. Do different ARs modulate LTCCs uniquely in the olfactory system? Does LTCC activity change accordingly with age? Is LTCC-dependent AHP an important player in olfactory learning? If so, how does NE modulate LTCC-dependent AHP? Does the age dependent dichotomy in β-adrenergic modulation of LTCC function in early life play a similar role in senescent plasticity as well? Future studies should be directed towards these questions to bring about a clearer understanding of the diverse roles provided by the crucial players in learning and memory.In the olfactory system, a concerted effort by NE, NMDARs and LTCCs in both the MOB (Jerome et al., AG, QY and XC developed the idea. AG did the literature review. AG and SJC wrote the manuscript. Further editing was done by AG, SJC, XC and QY.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.


OASL early post infection, where it functions in a prosurvival fashion by inhibiting autophagic mechanisms and antimicrobial peptide expression. This suggests an underestimated role of OASL in the innate immune response to infection with a variety of pathogens and points to OASL-associated modulation of the type I IFN response. OASL may therefore play a critical role in defining the outcome of infection. We provide a brief update on the recent developments of the OAS family of proteins in response to DNA and RNA virus infections, as well as discuss evidence of Oasl expression in response to a number of cytosolic and vacuolar replicating bacterial pathogens.The type I IFN response quickly became associated with its role in the innate immune response to viral infection. The past few years have seen the significance of IFNs expand in breadth to include non-viral pathogens. Previous work has identified that following viral infection, type I IFN signaling induces the production of the 2′-5′-oligoadenylate synthetase (OAS) family, which include OAS1, OAS2, OAS3, and OAS-like (OASL) protein. OASL was identified to be strongly induced following viral infection through engaging the RNA sensor RIG-I and increasing signaling through this pathway to enhance the anti-viral type I IFN response. Surprisingly, infection with viral dsDNA revealed an IFN inhibitory role and therefore pro-viral function of OASL through the inhibition of the cGAS cytosolic DNA sensing mechanism. Intracellular bacteria are able to activate the cytosolic DNA sensing pathway, however the role of OASL during bacterial infection is largely unknown. Vacuolar pathogenic microbes such as mycobacteria induce  Oasl expression in response to a number of intracellular bacterial pathogens.The 2′-5′-oligoadenylate synthetases (OAS) were the first interferon (IFN)-induced antiviral proteins described that are regulated by both type I and II IFNs  synthase (cGAS) is a sensor of cytoplasmic DNA, usually of microbial origin and has recently been thoroughly reviewed   are central to distinguishing these signals and sensing conserved motifs presented by microbes Gordon, . The memSting/Tbk1/Irf3 axis which will culminate in type I IFN secretion. It is now known that IFN-α IFN-β secretion in response to bacterial infection is in fact a pathogenic mechanism that enhances bacterial replication  microbe link, the next question that arises is whether OASL is relevant during infection with other vacuolar pathogens and those which replicate in the cytoplasm. A search of the literature revealed the upregulation of OASL or the murine equivalent Oasl2 through transcriptome analysis (Table Chlamydia trachomatis (Lad et al., Chlamydia pneumonia (Eickhoff et al., L. monocytogenes (Kutsch et al., Brucella abortus (Roux et al., Lactobacillus acidophilus (Weiss et al., OASL knock-down did not influence IFN-β release during M. leprae infection (de Toledo-Pinto et al., As with viral dsDNA, it was observed that the entry of foreign bacterial dsDNA is met with the DNA sensor cGAS, which then signals through cGAMP, activating the M. tuberculosis, M. leprae, and L. monocytogenes, and since OASL plays a significant role in modulating type I IFN production is can be said that the expression of this protein may be central to controlling viral and bacterial infections. Collectively, the above evidence clearly points to an underestimated role of OASL in the innate immune response to infection with a variety of pathogens. Not only is OASL expression closely related to the secretion of type I IFNs, its role is clearly diverse with it interacting closely with the RIG-I pathway, autophagy, and DNA sensing through cGAS.The balance between type I and type II IFNs is particularly important for controlling intracellular bacterial infections, notably GL wrote the manuscript. IW and BB provided substantial contributions to the framework of the manuscript, as well as revision of the work, and approval of the final draft.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.


Autism spectrum disorder (ASD) defines a group of neurodevelopmental disorders whose symptoms include impaired communication and social interaction with restricted or repetitive motor movements, frequently associated with general cognitive deficits. Although it is among the most severe chronic childhood disorders in terms of prevalence, morbidity, and impact to the society, no effective treatment for ASD is yet available, possibly because its neurobiological basis is not clearly understood hence specific drugs have not yet been developed. The endocannabinoid (EC) system represents a major neuromodulatory system involved in the regulation of emotional responses, behavioral reactivity to context, and social interaction. Furthermore, the EC system is also affected in conditions often present in subsets of patients diagnosed with ASD, such as seizures, anxiety, intellectual disabilities, and sleep pattern disturbances. Despite the indirect evidence suggestive of an involvement of the EC system in ASD, only a few studies have specifically addressed the role of the EC system in the context of ASD. This review describes the available data on the investigation of the presence of alterations of the EC system as well as the effects of its pharmacological manipulations in animal models of ASD-like behaviors. N-acyl-phosphatidylethanolamine phospholipase D (NAPE-PLD) whereas 2-AG is synthesized by diacylglycerol lipase (DAGL)-α and DAGL-β. Both ECs are rapidly degraded to terminate their actions: AEA is catabolized primarily by fatty acid amide hydrolase (FAAH) while the main 2-AG degrading enzyme is monoacylglycerol lipase (MAGL). ECs exert their effects mainly through the G protein-coupled cannabinoid receptors type 1 (CB1) and type 2 (CB2). Additionally, they are also able to interact with non-CB1/non-CB2 targets, including the transient receptor potential vanilloid type 1 (TRPV1) channel [Endocannabinoids (ECs) are arachidonic acid-derived compounds that, together with their receptors and the associated metabolic enzymes, constitute the endocannabinoid (EC) system, a neuromodulatory network of lipid signaling pathways . The mai channel  as well  channel .Alterations of the EC system functionality contribute to the pathogenesis of several psychiatric and neurological disorders ,6,7. IndDespite these observations being suggestive of an involvement of the EC system in ASD, only a few studies have specifically investigated the role of the EC system in the context of ASD, and our understanding of the EC signaling in ASD is still in its infancy.The following paragraphs will summarize animal data supporting the fact that (1) the EC system is altered in ASD and (2) pharmacological modulators  of the EAccording to the Fifth Edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5), ASD comprises a heterogeneous group of neurological conditions characterized by impaired social communication functions and the presence of restricted, repetitive patterns of behavior or interests that start to emerge in the early developmental period. In subsets of patients, common associated symptoms include seizures, anxiety, intellectual disabilities, motor dysfunctions, altered responsiveness to sensory stimuli, sleep, and metabolic disturbances . ExperimSeveral experimental manipulations based on important known causative genetic and environmental factors of ASD have been used to mimic ASD-like phenotypes in animal models. These include the disruption of genes responsible for syndromic forms of ASD, genetic manipulations mimicking human polymorphisms, as well as the exposure to known environmental risk factors. A detailed description of available animal models of ASD is beyond the scope of this review and therefore we refer the reader to recent and comprehensive reviews on this topic ,26,27. HFmr1 mutations lead to a developmental condition called Fragile X Syndrome (FXS). FXS is one of the leading monogenetic causes of ASD and is characterized by intellectual disabilities, developmental delays, anxiety, repetitive behaviors, and physical malformations . ApproxiThe first evidence supporting an involvement of the EC system in FXS came from the studies of Zhang and Alger  and MaccCollectively, these data suggest that alterations of the EC system may contribute to the symptoms of FXS and modulation of the EC signaling may have therapeutic potential in ameliorating some symptoms of FXS. Accordingly, enhancement of 2-AG signaling through administration of JZL184, a pharmacological inhibitor of the 2-AG degrading enzyme MAGL, corrected EC-dependent LTD and improved hyperlocomotion and anxiety-like responses in Fmr1 knockout mice .In addition, modulation of either CB1 or CB2 receptors was shown to ameliorate some of the behavioral signs of Fmr1 knockout mice . In partFinally, it has also been shown that modulation of AEA signaling can improve some aspects of the behavioral phenotype in Fmr1 knockout mice. Indeed, acute injection of the FAAH inhibitor URB597 improved aversive memory and anxiety-like behavior in Fmr1 knockout mice, without affecting social behavior . In contAs a whole, available data on Fmr1 knockout mice indicate that imbalanced EC signaling contributes to FXS and correcting EC signaling by targeting different components of the EC system  might offer an interesting therapeutic strategy for treating some of the clinical manifestations of FXS.NLGNs are postsynaptic cell adhesion molecules required for synaptic function; they orchestrate the maturation and function of both excitatory and inhibitory synapses in the mammalian brain ,47. In hThe presence of alterations of the EC system has been investigated both in the NLGN3R451C knockin and the NLGN3 knockout mouse models of ASD-like behaviors. In the hippocampus, both animal models showed a loss of tonic, but not phasic, CB1 receptor-dependent suppression of GABA release at synapses formed by cholecystokinin-expressing (CCK) basket cells to CA1 pyramidal neurons . In addiOverall, these findings suggest that NLGN3 is required for tonic EC signaling both in the hippocampus and in the somatosensory cortex. Given the common genetic association of the R451C substitution and NLGN3 deletion with ASD, it is possible that disrupted EC signaling could contribute to autism pathophysiology in these mouse models. However, specific pharmacological approaches aimed at modifying EC signaling in the brain of NLGN3 mutant mice are required to establish whether the observed alterations of the EC system directly contribute to the ASD-like phenotype in these mouse models.In addition to the genetically modified rodent models of ASD, several inbred mouse strains incorporate face validity as ASD models, because they display robust and well-replicated social deficits and repetitive behaviors. These inbred strains are considered to be models of idiopathic autism, as their ASD-like phenotype is not caused by known genetic mutations. Among these, of particular interest is the BTBR mouse, which has been the most extensively characterized. BTBR mice display deficits in behavioral tasks modeling the core domains of ASD, including reduced play and social approach behavior ,55 and rThus far, only one study has investigated the effects of a manipulation of the EC system in the BTBR mouse model . The autAlthough the most commonly suggested etiology of ASD is through the hereditary genetic characteristics identified as high risk genes for ASD, exposure to environmental factors in the prenatal and early postnatal periods imposes a significant contribution to ASD development ,61. ReleThe EC system has been studied in two environmental-based models: prenatal VPA exposure and postnatal lipopolysaccharide (LPS) administration in rats.Among environmental animal models that show both construct and face validity to ASD, the VPA rat model represents an excellent system to test and develop novel behavioral and drug therapies. This model demonstrates many of the structural and behavioral features that can be observed in individuals with autism, thus enabling definition of relevant pathways of developmental dysregulation resulting from environmental manipulation . VPA is The VPA rat model has been extensively used to evaluate the possible implication of the EC system in ASD. Adolescent rats exposed to a single VPA injection at gestational day (GD) 12.5 exhibited reduced mRNA expression of the enzyme primarily responsible for the synthesis of 2-AG DAGLα in the cerebellum and increased activity of the 2-AG-catabolising enzyme MAGL in the hippocampus . mRNA exOf note, enhancing AEA signaling through inhibition of its degradation partially mitigated the behavioral phenotype induced by prenatal VPA exposure. Indeed, systemic administration of the FAAH inhibitor PF3845 at the dose of 10 mg/kg attenuated the deficit in social behavior observed in VPA exposed male animals . In contOverall, available data in the VPA rat model suggest that altered AEA-mediated signaling may contribute to communication and social deficits associated with ASD, and support a role of FAAH in the regulation of social behavioral deficits.Both viral and bacterial infections during pregnancy have been linked to an increased risk to develop ASD in the offspring . ExposurAlthough post-natal LPS administration is not recognized as a model of ASD, data collected further suggest that FAAH inhibition may represent a potential approach for the treatment of disorders involving impaired sociability. Alterations of the EC system have been reported in rats after postnatal LPS exposure . Early-lMany morphological and neurochemical abnormalities have been reported in ASD patients as well as in animal models, reflecting the heterogeneous and complex nature of this group of disorders. Such diversity poses a great challenge and hampers the identification of possible common pathophysiological mechanisms in ASD.The EC system is involved in the modulation of many of the cellular functions and molecular pathways altered in ASD, such as imbalanced GABAergic and glutamatergic transmission, oxidative stress, altered energy metabolism, immune dysregulation ,88,89,90For instance, the reported prosocial effects of AEA in animal models of ASD might arise from an interaction with oxytocin, a neuropeptide that promotes parental and social bonding. Indeed, recent evidence has highlighted that oxytocin stimulates AEA release in the nucleus accumbens, a key region for the reinforcing properties of both natural rewards and drugs of abuse, and, importantly, AEA-mediated signaling is required for the prosocial effects of this neuropeptide . This suAdditionally, it has been shown that the amelioration of cognitive deficits after chronic rimonabant administration in Fmr1 knockout mice was associated with the normalization of the hippocampal mTOR signaling pathway , suggestThere is also evidence that ECs might modulate ASD symptoms via interaction with immune system cells. Indeed, changes in AEA metabolism and CB2 receptors were observed in peripheral blood mononuclear cells  and blooOf note, converging evidence indicates that ASD is characterized by an excitation/inhibition (E/I) imbalance in selective neuronal circuits that appears to be mainly related to defects of GABAergic signaling in different brain structures ,98. As rAlthough the preclinical findings seem to suggest that pharmacological interventions aimed at modulating the EC system could be beneficial for relieving symptoms associated with ASD , their pConverging data indicate that enhancing AEA signaling through inhibition of its degradation exerts prosocial effects in different animal models of ASD. In addition, CB1 receptor blockade, either acute or chronic, seems to have beneficial effects towards cognitive deficits, at least in mouse models of FXS. Remarkably, in most of the studies, the drugs were administered systemically. However, the alterations of the EC system reported in animal models of ASD  appear t


As expected, doubly heterozygous mice demonstrated GFP expression in most of the cells in the bone marrow and to a lesser extent in the spleen and thymus . The development of marginal zone (MZ) B-cells in the spleen relies on Notch 2 signaling and Notch blockade results in the reduction of the MZ B-cell pool in murine spleens.7 As shown in f/−Vav+/− mice compared to controls at 6 months. Second, we confirmed that these mice exhibited the expected abnormalities in thymocyte development, a Notch1 process.9 We found a significant increase in the double negative (DN) population within the GFP+ fraction of thymocytes taken from DNMf/−Vav+/− mice compared to controls  at the expense of an increase in granulocyte–monocyte progenitors (GMPs).genitors , a signigenitors  and a sigenitors . There wgenitors  or the Cgenitors . This sigenitors . Finallygenitors  compared5 To determine if canonical Notch signaling can function as a tumor suppressor in myelopoiesis, a tumor watch was established . The mice were aged to 15–18 months and there was no evidence of a highly penetrant myeloid disease with loss of canonical Notch signaling as shown by no significant difference in survival  but viable cells could not be obtained and the other died abruptly without splenomegaly. A single mouse in the control group also died abruptly without splenomegaly. These mice continued to show >90% GFP expression at 12 months and decreased MZ B-cells at 18 months  in control and DNMf/−Vav+/− lineage negative bone marrow cells followed by transplantation into syngeneic recipients , resulted in a relative increase in GMPs that was stable over time and an increase in myeloid cells in the spleen. Despite these observations, there were no overt clinical features consistent with the development of a myeloid neoplasm. Our results are consistent with other studies that evaluated the impact of Notch inhibition on normal hematopoiesis via expression of DN-MAML1 or conditional ablation of the Rbpj gene.4 Additionally, our results regarding the impact of Notch blockade on leukemia growth using the FLT3ITD model is congruent with our previous data, which demonstrated that inhibition of Notch signaling, via retroviral expression of DN-MAML1 or treatment with gamma secretase inhibitors, can inhibit leukemia growth.6 Furthermore, inactivating mutations in the Notch pathway are uncommon in primary myeloid neoplasms.12 These studies are in contrast to the results from disruption of Nicastrin, which led to a marked expansion of myeloid cells and a fully penetrant myeloid neoplasm at 5 months, prompting speculation that Notch functions as a tumor suppressor in myeloid cells.5The results from our study demonstrate that prolonged loss of Notch signaling in vivo, we utilized the dominant-negative mastermind-like transgene in which the ICN transcriptional co-activator MAML1 is truncated and fused to GFP7 preventing the formation of the ternary complex that is required for the transcriptional activation of Notch target genes. This approach should yield a more specific disruption of canonical Notch signaling than deleting Nicastrin, which has other substrates besides Notch. Finally, we avoided the Mx-1 cre system, and relied on the more hematopoietic-specific Vav-cre model. Not only is the Mx-1-Cre transgene active in mesenchymal stem cells in the bone marrow stroma,13 but it requires an interferon response for induction and it’s well known that interferon signaling can alter stem cell function and induce proliferation of myeloid cells.14 Collectively, these features may lead to cell extrinsic effects on myeloid development and leukemogenesis.These differences are likely multifactorial and include cell context, the varying contribution of intrinsic and extrinsic signaling and dosage of pathway modulation; altogether highlighting the complexity of the Notch pathway. To achieve loss of Notch1-4 signaling in vivo is insufficient for the development of myeloid neoplasms. This lack of a strong phenotype observed after Notch blockade on normal hematopoiesis is encouraging for clinical studies evaluating the impact of different Notch pathway inhibitors on both solid and liquid tumors.15 Furthermore, our results highlight the importance of using selective in vivo mouse models of Notch perturbation for future evaluation of the underlying mechanisms of Notch signaling in myeloid malignancies.Altogether, our results demonstrate that loss of Notch signaling


Moreover, extracellular Ca2+ levels impact on distinct processes in mammalian reproduction. The calcium-sensing receptor (CaSR) translates changes in extracellular Ca2+-concentrations into cellular reactions. This review summarizes current knowledge on the expression of CaSR and its putative functions in reproductive organs. CaSR was detected in placental cells mediating materno-fetal Ca2+-transport such as the murine intraplacental yolk sac (IPYS) and the human syncytiotrophoblast. As shown in casr knock-out mice, ablation of CaSR downregulates transplacental Ca2+-transport. Receptor expression was reported in human and rat ovarian surface epithelial (ROSE) cells, where CaSR activation stimulates cell proliferation. In follicles of various species a role of CaSR activation in oocyte maturation was suggested. Based on studies in avian follicles, the activation of CaSR expressed in granulosa cells may support the survival of follicles after their selection. CaSR in rat and equine sperms was functionally linked to sperm motility and sperm capacitation. Implantation involves complex interactions between the blastocyst and the uterine epithelium. During early pregnancy, CaSR expression at the implantation site as well as in decidual cells indicates that CaSR is important for blastocyst implantation and decidualization in the rat uterus. Localization of CaSR in human extravillous cytotrophoblasts suggests a role of CaSR in placentation. Overall, evidence for functional involvement of CaSR in physiologic mammalian reproductive processes exists. Moreover, several studies reported altered expression of CaSR in cells of reproductive tissues under pathologic conditions. However, in many tissues we still lack knowledge on physiological ligands activating CaSR, CaSR-linked G-proteins, activated intracellular signaling pathway, and functional relevance of CaSR activation. Clearly, more work is required in the future to decode the complex physiologic and pathophysiologic relationship of CaSR and the mammalian reproductive system.Active placental transport of maternal serum calcium (Ca Moreover, it is modulated by various physiological stimuli including extracellular pH, L-aromatic amino acids, and ionic strength. Different ligand binding sites can stabilize distinct conformational changes, which results in “ligand-biased signaling.” As a consequence, CaSR was shown to interact with the heterotrimeric G proteins Gq/11, Gi/o, G12/13, and Gs and is able to target all major intracellular signaling pathways. CaSR-mediated activation of Gq/11 results in stimulation of phospholipase C (PLC), Cai/o can inhibit adenylyl cyclase (AC). On the other hand, it can activate mitogen-activated protein kinases (MAPK) such as ERK1/2 and JNK. This can lead to transactivation of the epidermal growth factor receptor (EGFR). Activation of G12/13 modulates several pathways. This can lead to migration via rho-mediated actin polymerization and membrane ruffling or induce cell differentiation. It can also target tyrosine kinases, protein phosphatases, or activate certain AC isoforms. CaSR-coupling to Gs also activates ACs. Furthermore, CaSR activation can stimulate PLA, phosphatidylinositol 3-kinase (PI-3K) and PI-4K. Overall, major consequences of CaSR activation in cells are CaCaSR is, however, promiscuous in ligand binding; it can be stimulated by other divalent  activity, which stimulates expression of Ca2+-channels with appropriate response elements. And finally, CaSR can increase K+-channel activity, and as a consequence increase membrane potential and a Ca2+-driving force. Many intracellular signaling pathways, in turn, are sensitive to cytosolic Ca2+-rises, and as a consequence, cellular processes such as proliferation or migration can result from CaSR activation  or as a tumor suppressor gene  , mainly activation of MAPK pathways  has been investigated so far, probably because MAPK activation is of great importance in reproduction  the proliferation of the stem cells (spermatogonia) resulting in spermatocytes, (2) two meiotic divisions that give rise to haploid spermatids, and (3) spermiogenesis, the differentiation into mature sperms , while a CaSR agonist (AC-265347) inhibited PY similarly to CaIn isolated equine sperms, CaSR protein was located predominantly in the head of the sperm, with lower expression seen along the tail. Proteins detected with an anti-CaSR antibody by western blotting had a molecular weight of 100 and 75 kDa . It was concluded that CaSR activation had a pro-apoptotic impact on germ cells in diabetic rats and overall participated in diabetes-induced testicular damage.Diabetes, being an increasing problem worldwideWhile CaSR expression in normal rat Leydig cells could not be shown  receptor present on the surface of murine cells inducing Ca2+ release from ryanodine receptor-gated intracellular Ca2+ stores  is characterized by hyperandrogenism, oligo- and/or anovulation, and polycystic ovarian morphology. PCOS is associated with insulin resistance, hyperinsulinemia and central obesity. Excessive proliferation of theca cells is often associated with PCOS; this is a major cause of infertility due to ovarian hyperandrogenism  first proliferate and then enter the first meiotic division, which is halted in a prophase state of meiosis I until sexual maturity. This first meiotic arrest is characterized by a large nucleus called the germinal vesicle  from the pituitary gland, which stimulates the release of the oocyte from the ovary (ovulation). Upon that stimulus, meiosis I continues and is terminated shortly before ovulation. Oocytes at metaphase I (MI) stage are defined as oocytes with no GV (intact nucleus) and no polar body . For transition of the unfertilized egg to a developing embryo, egg activation is initiated by alterations in Ca2+ concentrations in the surroundings. Induced proliferation of these cells due to high Ca2+ concentrations may be of physiological relevance during the healing of the ruptured surface following each ovulation, when neighboring cells are locally exposed to ovarian follicular fluid. At least in pig ovarian follicular fluid, Ca2+ concentrations of around 2.34 mmol/L have been measured  cells. There, inositol triphosphate production increased after stimulation with Gd3+ and Ca2+. Expression of an interfering mutant CaSR inhibited the proliferative response to elevated extracellular Ca2+. CaSR agonists induced tyrosine phosphorylation, ERK activation and proliferation. Expression of interfering mutants for Ras, Raf, and MKK1 indicated that proliferation of ROSE cells in response to increased CaExpression of CaSR in the ovary was first described in human surface epithelial cells Figure . ProteinOvarian surface epithelial cells are the cell type primarily responsible for malignant ovarian carcinoma, which is the most lethal female genital malignancy. McNeil et al.  observed2+ in mobilization of intracellular Ca2+ during oocyte maturation, activation, and fertilization  and was not observed at suboptimal external Ca2+(0.5 mM). However, variations of Ca2+ between 0.5 and 4 mM Ca2+ in the absence of the agonist did not stimulate in vitro oocyte maturation. In oocytes treated with NPS R-467, CaSR immunostaining increased at the plasma membrane, while it was reduced in the cytosol. Finally, treatment of oocytes as well as cumulus cells with CaSR agonist resulted in an increased activity (phosphorylation) of MAPK (ERK) in these cells . Increasing extracellular Ca2+ resulted in a biphasic response of the cells; an initial inhibitory effect on apoptosis was followed by a delayed phase of increased apoptosis. As the initial inhibitory effect of the Ca2+ on apoptosis was mimicked by applying the CaSR agonists Mg2+ and Gd3+, an involvement of CaSR in inhibition of apoptosis was suggested  as well as the ions Ca2+ and Mg2+. The authors suggested that activated CaSR may play a role in securing the survival of avian follicles after their selection . Current data support a role of CaSR in maturation (completion of first meiosis) of oocytes and a contribution of the MAPK ERK signaling pathway. However, the interplay of CaSR with other molecules that maintain oocytes in the meiotic arrest and those that initiate meiotic resumption (such as LH) , to house the growing conceptus and to expulse the fetus during delivery . If communication between the embryo and the endometrium fails, then implantation fails. This is an important cause of infertility. There are estimations that only 50–60% of all conceptions advance beyond 20 weeks of gestation and from the pregnancies that are lost, 75% represent a failure of implantation. Failed implantation is also a major limiting factor in assisted reproduction. To treat disorders such as infertility, which increases worldwide due to an increasing toxic environment, but also diseases such as obesity, it is important to understand the molecular mechanisms of implantation and placentation  placenta formation, two distinct populations of trophoblast cells, the villous and the extravillous trophoblast, which exhibit distinct functions, become evident. The expanding chorionic villi of the placenta, which mediate materno-fetal transport of nutrients are covered by a multinucleated layer of syncytiotrophoblast (STB). This cell layer is maintained by proliferation and fusion of the underlying villous cytotrophoblasts. Extravillous trophoblasts, in contrast, invade the uterus and interact with maternal cells. These cells are important for proper placentation.invasive as seen in humans, most primates, dog, cat mouse, rat, or rabbit meaning that the conceptus will break through the surface epithel of the maternal uterus and invades the underlying stroma or can be non-invasive , integrating the uterine epithelium in the placenta. The depths of invasion as well as the degree of proximity between maternal and fetal circulation can vary largely among species. In humans and guinea pigs, the conceptus invades the stroma so deeply that the uterine surface epithelium is restored over it . Other species , may invade the stroma only partially and project into the uterine lumen (eccentric implantation). This can result in a contact to other sites of the uterine lumen and additional placental development . While in some species the maternal tissue remains relatively intact , meaning that trophoblast cells contact maternal capillary endothelium, in other species, trophoblast cells also invade the maternal endothelium and ultimately bath in maternal blood . These types of placentas are classified as haemochorial, which means that all maternal tissue layers are removed and the chorion  bathes directly in maternal blood. But even structures of murine and human placentas differ to some extent. Whereas, the term human placenta is monochorial, which means that one continuous trophoblast cell layer, the STB, separates maternal and fetal blood circulation, the murine placenta is trichorial. Moreover, in rodent placenta, but not in human, other primate, or ruminant placentas the primitive yolk sac, which in all species participates in nutrient exchange between the fetal and maternal circulations before the formation of the placenta, is incorporated into the placenta and turns into the intraplacental yolk sac (IPYS). The IPYS is positioned between fetal vessels and maternal blood spaces, well situated for exchange of substances between mother and fetus. It is a bilayered membrane, where smaller parietal or cuboidal cells on a thick basement membrane (Reichert's membrane) overlie maternal blood spaces and vessels, while tall columnar cells on the visceral or endothelial side overly the fetal vessels. Between these layers is the sinus of Duval, which communicates with the yolk sac cavity and the uterine lumen . Current knowledge on placental Ca2+ transfer as well as regulation of maternal and fetal Ca2+ homeostasis have been reviewed in several recent publications  to stimulate Ca2+ secretion into milk when the Ca2+ supply is adequate  by immunization of mice with intact parathyroid cells. Two of these antibodies (G11 and B6) interfered with the Ca2+-sensing mechanism of parathyroid cells .The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cystic kidney diseases comprise a varied collection of hereditary disorders, where renal cysts comprise a major element of their pleiotropic phenotype. In pediatric patients, the term polycystic kidney disease (PKD) commonly refers to two specific hereditary diseases, autosomal recessive polycystic kidney disease (ARPKD) and autosomal dominant polycystic kidney disease (ADPKD). Remarkable progress has been made in understanding the complex molecular and cellular mechanisms of renal cyst formation in ARPKD and ADPKD. One of the most important discoveries is that both the genes and proteins products of ARPKD and ADPKD interact in a complex network of genetic and functional interactions. These interactions and the shared phenotypic abnormalities of ARPKD and ADPKD, the “cystic phenotypes” suggest that many of the therapies developed and tested for ADPKD may be effective in ARPKD as well. Successful therapeutic interventions for childhood PKD will, therefore, be guided by knowledge of these molecular interactions, as well as a number of clinical parameters, such as the stage of the disease and the rate of disease progression. PKHD1 gene which encodes a protein known as fibrocystin or polyductin (FPC), and both the gene and protein interact with the autosomal dominant polycystic kidney disease (ADPKD) genes and proteins.Autosomal recessive polycystic kidney disease OMIM #263200) is characterized by renal cysts and hepatobiliary dysgenesis and is a substantial cause of morbidity and mortality in children , 2. ARPK63200 is Autosomal dominant polycystic kidney disease is one of the most common genetic disease affecting 1/400 to 1/1,000 individuals worldwide. ADPKD is generally a late-onset, systemic disease characterized by bilateral, progressive enlargement of focal cysts occurring in all nephron segments with variable extrarenal manifestations .PKD1 (on chromosome 16p13.3) (OMIM #173900) and PKD2 (on chromosome 4q21) (OMIM #173910). These genes encode the protein polycystin 1 (PC1) and polycystin 2 (PC2), respectively.Autosomal dominant polycystic kidney disease was originally thought to be caused by mutations in two genes; in GANAB (OMIM 104160) encoding the glucosidase II subunit α on chromosome 11q12.3 cause a mild form of ADPKD and autosomal dominant polycystic liver disease of varying severity, most likely due to defects in PC1 maturation  that did not link to either locus –6. Recenturation .Childhood ADPKD may be indistinguishable from ARPKD, and histological or genetic analysis may be necessary to differentiate the two , 8. The Cyst formation and progressive growth is a complex dynamic process with multiple interacting signaling components that all contribute to disease, but never act autonomously. The early investigations of PKD focused on fundamental phenotypic changes that would be necessary for a normal renal tubular epithelial cell to become a cystic epithelial cell. Normal renal epithelial cells changed from a mature, differentiated, non-proliferative, absorptive cell to a partially de-differentiated secretory cell characterized by specific polarity defects and increased rates of proliferation .These led to the identification of a myriad of signaling molecules and signaling pathways that were found to be abnormal in cystic epithelium. Collectively, these changes define what is referred to as the “cystic phenotype.” Figure In vitro and in vivo experimental models have been used to identify fundamental pathogenic features in human PKD and are thought to be pathogenic in both ARPKD and ADPKD. These include the following:aberrant intracellular levels of the second messenger, cyclic adenosine monophosphate (cAMP), coupled with decreased intracellular calcium increasing both proliferation and secretion;abnormalities in expression and localization of the ErbB or epidermal growth factor (EGFR)—family of receptors and ligands (EGFR axis), leading to increased proliferation;abnormal activity of cSrc (p60Src), a non-receptor tyrosine kinase that serves as a critical mediator of cross talk between the EGFR axis and G-protein-cAMP pathways. In addition, cSrc interactions with the cleaved C-terminal PC1 tail (PC1-p30) activate the transcription factor STAT3;activation of mammalian target of rapamycin (mTOR) signaling;alterations in cell–cell and cell–matrix interactions;changes in interstitial macrophages that leads to progressive interstitial fibrosis.http://clinicaltrials.gov/.Clinical trials have to date been largely based on targeting these complex cellular signaling features of a cystic cell. Therefore, a brief discussion of each and the evidence supporting the pathogenic features listed above are included below. A brief discussion of the results of clinical trials to date will follow. A list of current and completed clinical trials can be found at The cAMP-dependent pathway is a G-protein-coupled receptor signaling pathway. Increased intracellular cAMP levels in a normal renal epithelial cell elicit a response that reduces MAPK activity resulting in a decreased rate of cellular proliferation. However, under conditions of low levels of intracellular calcium, a cSrc-dependent phosphorylation of β-Raf, allows the cell to bypass Raf-1and increase ERK phosphorylation and subsequent cell proliferation , that inhibit adenylate cyclase resulting in reduced cAMP levels , 14.®) has been tested in human patients with ADPKD and the results have been encouraging. Tolvaptan slowed the increased in total kidney volume and slowed the decline in loss of renal function  and other ErbB receptors and their ligands (the EGFR axis) are important mediators of renal cystic epithelial proliferation in PKD. In human ADPKD and ARPKD and rodent models of PKD, renal cystic epithelia display overexpression and mislocalization of one or more ErbB receptors to the apical cell surface instead of the customary basolateral localization seen in the normal human kidneys –19. Thesin vitro . Preclinhibitors , 22 or ghibitors  and/or rhibitors  significCre;del2-11,loxPkd1) found that ErbB4 activation was a major driver of cellular proliferation in ADPKD and may well be a biomarker of disease progression  the EGF receptor resulting in a reciprocal phosphorylation (activation) of cSrc. Phosphorylated cSrc activates β-Raf, which alters the response of renal epithelia to cAMP from a normally antimitotic to a pro-proliferative phenotype. Additionally, activated cSrc interacts with PC1-p30, a proteolytic fragment of the PC1 cytoplasmic tail, resulting in STAT3 phosphorylation and further increased proliferation , G-protein coupled receptors (which generate cAMP), cellular energy levels, nutrient status, and stress conditions to stimulate protein synthesis and cell growth through activation (phosphorylation) of S6K1 and eIF4E , 32. In It is impossible to imagine how a single compound could provide lifetime effective therapy even if started early in the disease process. However, the identification of pathological cellular events provides a starting point for building future therapeutic interventional strategies for childhood PKD. Challenges including the focal nature of cyst formation and the large variation in the phenotypic expression of ARPKD and ADPKD are not trivial. The substantial intra-familial phenotypic variability seen in both ARPKD and ADPKD suggests that complex factors which influence or direct the timing and severity of disease are operative.Pkd1, Pkd2, and Pkhd1. Studies in such models which more accurately reflect the human disease have yielded unexpected results regarding mechanisms of cyst development and enlargement in PKD. Advances in sequencing, in concert with improved methods to produce animal models with specific mutations including those found in PKD patients, allow suspected mechanistic processes to be directly tested. These new insights along with lessons learned from the original clinical trials will lead to novel therapeutic interventions for PKD.Improved molecular techniques and increased specificity in producing targeted gene mutations has allowed development of orthologous rodent models with conditional and hypomorphic mutations in Pkd1, Pkd2, Pkhd1, and the two polycystic liver disease genes, Prkcsh and Sec63, resulted in different combinations of mutant alleles which allowed the functional relationships between these genes to be defined  PC1 dosage modifies the severity of both ADPKD and ARPKD; (3) the threshold level of PC1 necessary for normal tubular morphology varies by nephron segment with collecting ducts being most sensitive; and (4) overexpression of Pkd1 is capable of rescuing a mutant Pkhd1 animal  cell proliferation; (b) cAMP levels; and (c) fluid secretion. Proliferation is an absolute necessity for cyst formation and as such, it must be targeted and controlled for any therapy to be effective; how much and for how long remains unknown.In the near future, the most promising therapies will target key signaling intermediates that integrate multiple pathways, such as cSrc  when therapy begins and the rate of progression, will require multiple agents or a single agent that hits multiple targets, and the choice of targets will be stage specific and change as disease progresses.There are a number of additional important factors to consider in developing therapies for childhood PKD. Therapies will increasingly focus on treating PKD in its earliest stages where they are likely to offer the maximum long-term benefit. Therapies that target abnormal proliferative pathways will need to be carefully monitored so that pathway activity is reduced to normal basal levels rather than completely shut off. Promising compounds may be modified to direct the molecule to the site of disease, making these therapies highly specific with low levels of toxicity . This sp® were reported to have increased urinary shedding of heparin-binding EGF-like growth factor (HB-EGF)  . In ARPKThe progress made in understanding the pathophysiology of PKD has been remarkable and despite the work still remaining, patients and parents can take heart that two clinical trials for children with PKD are underway: one for children with ADPKD [Tolvaptan (NCT02964273) Belgium and Italy] and one recently approved for the use of multi-kinase inhibitor, TSV in children with ARPKD.WS and EA contributed equally to the conception and design of the manuscript; drafted the article and made critical revisions related to the intellectual content of the manuscript; and approved the final version of the article to be published.The authors declare that they have no conflicts of interest with respect to research, authorship and/or publication of this manuscript.


Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive loss of lower and upper motor neurons (MN) leading to muscle weakness, paralysis and eventually death. Although a highly varied etiology results in ALS, it broadly manifests itself as sporadic and familial forms that have evident similarities in clinical symptoms and disease progression. There is a tremendous amount of knowledge on molecular mechanisms leading to loss of MNs and neuromuscular junctions (NMJ) as major determinants of disease onset, severity and progression in ALS. Specifically, two main opposing hypotheses, the dying forward and dying back phenomena, exist to account for NMJ denervation. The former hypothesis proposes that the earliest degeneration occurs at the central MNs and proceeds to the NMJ, whereas in the latter, the peripheral NMJ is the site of precipitating degeneration progressing backwards to the MN cell body. A large body of literature strongly indicates a role for the immune system in disease onset and progression via regulatory involvement at the level of both the central and peripheral nervous systems (CNS and PNS). In this review, we discuss the earliest reported immune responses with an emphasis on newly identified immune players in mutant superoxide dismutase 1 (mSOD1) transgenic mice, the gold standard mouse model for ALS. Motor neuron diseases are devastating and encompass a variety of disorders that lead to death of motor neurons and resulting loss of motor function, dysphagia and respiratory failure. Of these diseases, amyotrophic lateral sclerosis (ALS) is the most common. ALS frequently afflicts adults in middle age (40–60 years old) with an average lifespan of 3–5 years following diagnosis . Most caThere is currently a delay of more than one year between symptom onset and clinical diagnosis, likely due to the underestimation of initial symptoms by patients and clinicians as they are similar to those resulting from various etiologies . ALS is Research into the immune system influence in ALS has yielded important clues on possible markers that could be targeted to tackle the disease before it has progressed too far. Much research into immune system involvement in different stages of ALS disease has been covered to date ,7. HowevIn addition to MN cell intrinsic molecular defects, secondary mechanisms including toxic glial microenvironment, immune system influence on glial activation and neuromuscular junction (NMJ) maintenance, all converge to modify the disease course in ALS . Here, wCD4−/− background that lack CD4+ T cells, a similar significant reduction in their lifespan is observed relative to mSOD1 mice. The mSOD1 CD4−/− mice also exhibit attenuated microglial and astrocyte reactivity. Lack of CD4+ T cells alters the glia phenotype towards toxicity, owing to reduced expression of neurotrophic factors such as IGF-1, GDNF, BDNF; anti-inflammatory cytokines such as IL-4, TGF-β and increased expression of proinflammatory molecules such as TNF-α, NOX2 in the lumbar spinal cords of these mice . T. T104]. Despite our expanding understanding of cellular and physiological contributors to ALS onset and progression, there is still much to learn and identifying such processes and stages in human ALS is difficult. Therefore, improving our ability for early detection of the disease by identifying specific biomarkers that indicate temporospatial state of disease and immunologic and inflammatory correlates with these markers is essential for the best assessment of the efficacy of the variety of clinically-approved and newly developed immune and inflammation-targeted therapies. Also, new animal models that more widely represent the true ALS patient population beyond mutant SOD1 and other hereditary minority subsets of the disease would likely aid in screening such therapies before advancing to human application.Though recent research has expanded our understanding of the role of the immune response in MN diseases such as ALS, this review highlights not only new findings that contribute to this understanding, but also gaps in knowledge that still remain. ALS is considered a non-cell autonomous disease, as we now know many non-neuronal nervous system cells contribute to pathology, including astrocytes, microglia and neuromuscular interactions. We have focused on highlighting how complex immune system involvement may be in predicting the motor function outcome in this debilitating disease. It will be key for future research to connect findings involving immune involvement in CNS cell responses to those that reside in the peripheral segment: Schwann cells, fibroblasts, muscle cells, etc. Such investigations hold tremendous promise in identifying patterns to characterize immune responses at different disease phases that lead not only to potential therapeutic targets, but also biomarkers to improve speed of diagnosis.