Carotenoids found in these animals provide the food chain as well as metabolic pathways. In the present review, I will describe marine animal carotenoids from natural product chemistry, metabolism, food chain, and chemosystematic viewpoints, and also describe new structural carotenoids isolated from marine animals over the last decade. Marine animals contain various carotenoids that show structural diversity. These marine animals accumulate carotenoids from foods such as algae and other animals and modify them through metabolic reactions. Many of the carotenoids present in marine animals are metabolites of β-carotene, fucoxanthin, peridinin, diatoxanthin, alloxanthin, and astaxanthin,  Their astaxanthins consisted of three optical isomers and originated from dietary zooplankton .8) from β-carotene (52), ingested from dietary algae, via echinenone (53), 3-hydroxyechinenone (54), canthaxanthin (11), and adonirubin (12), as shown in Carotenoids in the carapace of crustaceans exist as both free and esterified forms. The principal carotenoid in crustaceans is astaxanthin ,7. In cr55) and lutein to fritschiellaxanthin (56) and papyrioerythrinone (57) [58) in the sea louse Ligia exotica exists as two optical isomers [59) [60) [Paralithodes brevipes and crawfish Procambarus clarkii, respectively (Some crustaceans can convert zeaxanthin to adonixanthin one 57) ,6,7. Cru ,6,7. Cr isomers ,6,7. Rec ,6,7. Crers [59)  and 2,3-59) [60) , were isectively .Z configuration (61) [Echinenone is a well-known major carotenoid in the gonads of sea urchins and is an oxidative metabolite of β-carotene ,7. Echinion (61) . 8), 7,8-didehydroastaxanthin (9), and 7,8,7′,8′-didehydroastaxanthin (10). They correspond to the oxidative metabolites of β-carotene, diatoxanthin, and alloxanthin, respectively. The crown-of-thorns starfish Acanthaster planci is a large, nocturnal sea star that preys upon coral polyps. Recently, four new carotenoids: 4-ketodeepoxyneoxanthin (62), 4-keto-4′-hydroxydiatoxanthin (63), 3′-epigobiusxanthin (64), and 7,8-dihydrodiadinoxanthin (65), were isolated from A.planci as minor components along with the major carotenoids 7,8-didehydroastaxanthin, peridininol, and astaxanthin, and several other minor carotenoids including 7,8,7′,8′-tetrahydroastaxanthin, diadinoxanthin, diatoxanthin, and alloxanthin [Starfish are carnivorous and mainly feed on bivalves and small crustaceans. Principal carotenoids in starfish are astaxanthin (oxanthin .66), was reported in the brittle star Ophioderma longicaudum [Z, 9′Z configurations, termed cucumariaxanthin (67), were isolated from the sea cucumber Cucumaria japonica [3,4,3′,4′-Tetrahydroxypirardixanthin 4,4′-disulfate, named ophioxanthin , an acetylenic analog of fucoxanthinol (26), and mytiloxanthinone (68), an oxidative metabolite of mytiloxanthin (28), were first isolated from the sea squirt Halocynthia roretzi [34) and amarouciaxanthin B (69), having a unique 3-oxo-6-hydroxy-ε-end group, were first isolated from the tunicate Amaroucium pliciferum [Halocynthiaxanthin  in muscle. Except for catfish, carotenoids in the integuments of fish exist in an esterified form.Many fish accumulate carotenoids in their integuments and gonads. On the other hand, Salmonidae fish peculiarly accumulate astaxanthin (8) is widely distributed in both marine and fresh water fish. Cyprinidae fish, which inhabit fresh water, can synthesize -astaxanthin (8a) from zeaxanthin (70) by oxidative metabolic conversion (Astaxanthin (nversion . On the axanthin 0 by oxid71) is widely distributed in fish belonging to Perciformes. The bright yellow color in the fins and skin of marine fish is due to the presence of tunaxanthin. Feeding experiments involving red sea bream and yellow tail revealed that tunaxanthin (71) was metabolized from astaxanthin (8) via zeaxanthin, as shown in 72) [Tunaxanthin 1 is widen in 72)  are key 73-76), were reported from the integuments of the black bass Micropterus salmoides [. They were assumed to be corresponding oxidative cleavage products of tunaxanthin, lutein, and alloxanthin. Unique apocarotenoids, micropteroxanthins , deepoxysalmoxanthin (78)  [79) (from the freshwater goby Rhinogobius brunneus) [Silurus asotus [80), 7′,8′-dihydrodiatoxanthin (81), and -ε-cryptoxanthin (82), were isolated from the integuments and gonads of the Japanese common catfish as minor carotenoids [Since 2000, there are a few reports on new structures of carotenoids from fish . Carotenus keta) , and gobrunneus) , were iss asotus . Recentlotenoids .There are few reports available on carotenoids in marine mammals. Only, β-carotene and lutein were reported from the dolphin . The wha25) in mice was investigated. Dietary administrated fucoxanthin was converted to amarouciaxanthin A (34) via fucoxanthinol (26) in mice [+ as a cofactor [Recently, absorption and metabolism of fucoxanthin  in some freshwater fish ,47.Many marine animals accumulate carotenoids in their integuments. Integumentary carotenoids may contribute to photoprotection, camouflage, and signaling such as breeding color. Carotenoids have excellent antioxidative activities for quenching singlet oxygen and inhibiting lipid peroxidation. Astaxanthin supplementation in Salmonidae fish suppressed oxidative stress ,49. Marine animals also accumulate carotenoids in their gonads. Carotenoids are assumed to be essential for reproduction in marine animals. Astaxanthin supplementation in cultured salmon and red sea bream increased ovary development, fertilization, hatching, and larval growth . In the Phaffia yeast and Haematococcus algae is widely used for the pigmentation of salmon, trout, and red sea bream. Lutein from marigold is also used as a yellow coloration for cultured marine fish such as yellow tail and red sea bream. Zeaxanthin from spirulina is used as a red coloration for goldfish and ornamental carp. Carotenoids are used for pigmentation in several aquaculture fish. Synthetic and natural astaxanthin from In the present review, I have described marine animal carotenoids from natural product chemistry, metabolism, food chain, and chemosystematic viewpoints and also describe new structural carotenoids isolated from marine animals during the last decade. In plants and photosynthetic bacteria, biosynthetic roots of carotenoids were identified at the enzymatic and gene level. On the other hand, neither enzymes nor genes for the metabolism of carotenoids in animals have been clarified. Therefore, chemical, biochemical, and analytical approaches are still important to clarify carotenoids in animals.Interesting new structural carotenoids can still be found in marine animals. The structures of these new carotenoids provide information on the function and food chain, as well as metabolic pathways in marine animals.


Nevertheless, most STAT3 inhibitors have yet to be translated to clinical evaluation for cancer treatment, presumably because of pharmacokinetic, efficacy, and safety issues. In fact, a major cause of failure of anticancer drug development is lack of efficacy. Genetic interactions among various cancer-related pathways often provide redundant input from parallel and/or cooperative pathways that drives and maintains survival environments for cancer cells, leading to low efficacy of single-target agents. Exploiting genetic interactions of STAT3 with other cancer-related pathways may provide molecular insight into mechanisms of cancer resistance to pathway-targeted therapies and strategies for development of more effective anticancer agents and treatment regimens. This review focuses on functional regulation of STAT3 activity; possible interactions of the STAT3, RAS, epidermal growth factor receptor, and reduction-oxidation pathways; and molecular mechanisms that modulate therapeutic efficacies of STAT3 inhibitors. Signal transducer and activator of transcription 3 (STAT3) plays critical roles in tumorigenesis and malignant evolution and has been intensively studied as a therapeutic target for cancer. A number of STAT3 inhibitors have been evaluated for their antitumor activity  Constit in vivo ,15,16, d in vivo , peptide in vivo , and sma in vivo ,20,21,22r models ,24,25,26r models ,28, inhir models . In addir models ,31 appror models ,33. Arser models ,35. Furtr models , inhibitr models ,38. HoweLack of therapeutic efficacy may be caused by low potency of the candidate drug in inhibiting its proposed target. Nevertheless, mutation analyses of primary cancer cells for genes encoding kinases or known to be associated with cancers have revealed that individual tumors may harbor multiple changes in such genes ,40,41,42C-terminal transactivation domain of STAT3α is deleted in STAT3β and replaced by seven unique C-terminal residues (CT7) whose functions remain undefined [C-terminus. The human STAT3 gene is located on chromosome 17q21.31 and encodes two major isoforms of STAT3 proteins via alternative mRNA splicing: STAT3α (p92) and STAT3β (p83). A 55-residue ndefined . Both is−/− mice that die at early stages of embryonic development [Targeted disruption of the murine Stat3 gene leads to early embryonic lethality. Stat3-deficient embryos have exhibited rapid degeneration from embryonic day 6.5 to embryonic day 7.5 . Ablatioelopment , expresselopment .De novo dominant-negative mutations in the DNA-binding domain of STAT3 have been identified in autosomal dominant or sporadic cases of hyperimmunoglobulinemia syndrome  in humans [De novo deficiency mutations of STAT3 have also occurred in the SH2 domain [n humans . These m2 domain ,53 and t2 domain  of STAT32 domain . These d2 domain ,57,58.There are only a few reports of STAT3 gene mutation in human cancer cells. In one report, a patient with HIES owing to STAT3 mutation had a subsequent primary parotid gland diffuse large B-cell lymphoma . BecauseGenetic interactions are functional cross-talks among genes, which regulate or compensate for one another in many signaling and/or metabolic pathways, leading to phenotypic changes, including disease status (synthetic sickness) and viability  alterations. Genetic interactions have been used by investigators to identify genes that are crucial to the survival of certain oncogene-transformed cells ,80,81,82STAT3 is activated by tyrosine phosphorylation in response to stimulation of a variety of cytokines, such as IL-6, leptin, prolactin, erythropoietin, and thrombopoietin, and growth factors, such as epidermal growth factor (EGF), fibroblast growth factor, insulin-like growth factor, hepatocyte growth factor, platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) ,94,95,96N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) may enhance or be required for STAT3/DNA interaction [The activity of STAT3 is also regulated by serine phosphorylation and lysine acetylation. Phosphorylation of S727 in STAT3 by c-Jun eraction ,103 and eraction ,104. Bloeraction . Nevertheraction ,107. Aceeraction ,109. Thieraction . A receneraction .Within cells, STAT3 activity is also regulated by several inhibitory molecules that prevent its continued activation. For example, tyrosine dephosphorylation of receptors and JAK receptor-associated phosphatases lead to inactivation of JAKs and prevent further STAT activation ,113,114.in vitro and tumor growth in vivo [in vitro and in vivo [The complexity of STAT3 activity regulation networks suggests that pharmaceutical inhibition of STAT3 activity can be achieved with a variety of mechanisms. Small-molecule inhibitors of upstream kinases JAK, Src, and Bcr-Abl are predicted to inhibit STAT3 activation. Indeed, the JAK inhibitors INCB16562, AZD1480, and tofacitinib (CP-690550) have potently blocked STAT3 signaling, suppressed cancer cell proliferation, or induced apoptosis  in vivo ,124,125. in vivo ,127,128  in vivo ,130, res in vivo . Phospho in vivo .As members of a subfamily of small guanine nucleotide-binding proteins, Ras proteins  cycle between active GTP-bound and inactive GDP-bound forms ,134. BinBoth Ras and STAT3 proteins are activated by EGFR ,141,147 Studies of K-ras-transgenic mice demonstrated that Stat3 phosphorylation occurs in tumors at multiple stages of K-ras-induced pancreatic tumorigenesis but not in normal pancreatic tissue ,166. DisBoth Ras and STAT3 regulate the activity of Rho-family small GTPases. STAT3 regulates Rac1 activity by interacting with the Rac1 activator betaPIX in the cytoplasm, thereby modulating the organization of the actin cytoskeleton and cell migration . Althougin vitro and in vivo. Similarly, investigators observed a synergistic effect between a PI3K inhibitor and a STAT3 inhibitor in human gastric cancer cells harboring KRAS mutations [The interaction between STAT3 and Ras signaling pathways suggests that the functional status of STAT3 affects responses of cancer to treatment with agents targeting these signaling pathways. Indeed, a study of response to treatment with the MEK inhibitor AZD6244 using a panel of lung cancer cell lines revealed that activation of the STAT3 pathway is associated with resistance to AZD6244 . Inhibitutations . Moreoveutations ,177 wereutations ,179, demEGFR-mediated signaling pathways are known to be a driving force in lung tumorigenesis and have been extensively investigated as targets for cancer therapy. Activating EGFR mutations are detected in about 10%–17% of human lung adenocarcinoma cases, with higher percentages in women and patients with no smoking history ,181,182.s [in vitro study using recombinant proteins demonstrated that STAT3α and STAT3β formed stable complexes with EGFR and were phosphorylated in tyrosine by the EGFR and activated for binding to DNA [STAT3 is identified because of its activation by tyrosine phosphorylation in response to exposure to EGF and IL-6 . Activats ,194. EGFs , upreguls ,197. An g to DNA . Activatg to DNA  and Grb2g to DNA ,200. In g to DNA ,201, wheg to DNA  and the g to DNA .in vitro. Inhibition of STAT3 expression by an antisense oligonucleotide dramatically suppressed the transforming growth factor-α/EGFR-mediated growth of transformed epithelial cells [As a key downstream mediator of the EGFR signaling pathways, STAT3 is crucial to EGFR-mediated cell growth al cells . STAT3 ial cells ,206. Moral cells , suggestal cells ,209 and al cells ,211  reactions, a molecule’s oxidation state changes because of a gain or loss of electrons. Inside a cell, redox metabolism is balanced by production and elimination of ROS, a group of oxygen- or nitrogen-containing molecules that are highly active in redox reactions. Produced in living organisms via a wide range of physiological process, ROS can serve as second messengers in response to exposure to growth factors, hormones, and cytokines. For example, Hytokines ,219. Alsytokines , and is ytokines , adhesioytokines , migratiytokines ,223, difytokines , oncogenytokines , and apoytokines ,227,228.ytokines ,230,231,ytokines , metastaytokines ,234,235,ytokines ,237, andytokines ,239. Nevnes [Ras , c-Myc [c [c-Abl , and SrcAbl [Src  and growAbl [Src , EGFR [2Abl [Src , and VEGAbl [Src  may rendAbl [Src ,246. IndAbl [Src ,248 and Abl [Src  induce a2O2 stimulates the activity of the STAT kinases JAK2 and TYK2 and activates STAT1 and STAT3 in fibroblasts, lymphocytes, and cancer cells. Activation of STATs by PDGF is markedly inhibited by the ROS scavenger N-acetyl-L-cysteine and diphenylene iodonium, indicating that ROS production contributes to STAT activation in response to PDGF exposure. These findings demonstrate that the JAK/STAT pathway responds to intracellular ROS induction and that PDGF uses ROS as second messengers to regulate STAT activation [2− anions generated by the NAD(P)H oxidase system [Cellular ROS may have either a stimulatory or inhibitory effect on STAT3 signaling depending on the cellular context or level or duration of ROS generation in a cell. Oxidative stress has been reported to stimulate the JAK/STAT pathway ,251,252.tivation . Stimulae system . ROS aree system . Increase system ,255,256.e system . Interese system , indicatin vitro, presumably via oxidation of crucial dithiols to disulfides in JAK2. The autokinase activity of JAK3 responds in a similar fashion to exposure to thiol redox reagents in vitro and nitric oxide donors in vivo. Treatment with parthenolide, an anti-inflammatory compound, increases intracellular ROS level and inhibits JAK1 and STAT3 activation but stimulates the MAPK pathways. Pretreatment with the antioxidant N-acetyl-L-cysteine completely suppressed the inhibitory effect of parthenolide on JAK1 and STAT3 [Evidence also suggests that ROS attenuate cytokine-induced JAK/STAT activation ,260,261.nd STAT3 . The cysnd STAT3 .In contrast to many reports on ROS-mediated changes in STAT3 activity, only a few reported studies have examined regulation of ROS levels by STAT3. In cardiomyocytes, constitutively active STAT3 protects against hypoxia- and/or reoxygenation-induced injury by scavenging ROS via upregulation of expression of manganese superoxide dismutase and its enzyme activity  and via Efficacy and toxicity are the two major issues in drug development. A survey of anticancer agents evaluated in human studies from 1991 to 2000 demonstrated that only about 5% of those entering clinical trials were approved for clinical use, and the majority of treatment failures in late-phase clinical trials of candidate anticancer agents resulted from a lack of efficacy of the drugs ,272,273.in vivo toxicity and efficacy profiles.The advances in knowledge about networks of genetic interactions are expected to impact strategies for enhancing the therapeutic efficacy of anticancer therapy. The interactions of the STAT3 pathway with other cancer-related pathways, such as the Ras, EGFR, and redox pathways, indicate that simultaneous targeting of several key molecules in these pathways using either combination therapy or agents capable of modulating the functions of multiple targets will be required for effective cancer therapy. Our own experience in anticancer drug development demonstrated that genetic interactions can be exploited for the development of anticancer agents targeting multiple pathways and that performing robust compound optimization once a lead compound is identified is critical, as compounds with similar chemical structures and in vitro activity may have dramatically different


This corrigendum/addendum supplies corrected statements and proofs of an error by author.Hear by, authors declare that the corrected authors list and affiliations are:1, Mohammad Kargar 1,*, Tarlan Jafarpour 1Akram Najafi 1 Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, IR IranSincerely YoursMohammad Kargar


Background. Significant decreases in p16 expression have been shown to occur in melanoma compared to Spitz tumors, and loss of p16 staining has been found to correlate with melanoma tumor progression. However, comparison of p16 between atypical cellular blue nevi (CBN) and melanoma has not been reported previously. Methods. p16 immunohistochemical staining was evaluated in 14 atypical CBN, 8 conventional and atypical melanocytic nevi, and 16 melanomas, including 4 malignant CBN. p16 staining intensity was graded on a scale of 0–3 and the percentage of melanocytes stained with p16 was determined. Results. p16 staining was significantly higher in all CBN as a group when compared to melanomas (P = 0.001) and malignant CBN (P = 0.00008). Higher p16 expression was also seen in mildly (P = 0.0002) and moderately atypical (P = 0.02), but not severely atypical, CBN compared to melanomas. Conclusions. p16 immunohistochemical expression is higher in mildly and moderately atypical CBN compared to severely atypical CBN and melanomas. In conjunction with additional markers and histology, p16 staining may be useful in confirming the benign nature of these tumors, but is not useful in distinguishing severely atypical CBN from malignant cases, consistent with the overlapping histologic features between these tumors. Distinguishing atypical and unusual variants of melanocytic nevi from melanoma by routine histologic examination can be difficult in some cases –3. In paCDKN2 [Immunohistochemistry is a useful tool in the diagnosis of some cases of melanoma, and markers such as S-100, HMB-45, Melan-A, MITF, and the proliferation marker Ki-67 are often used. Ki-67, in particular, has been found useful to distinguish benign from malignant melanocytic lesions [CDKN2 , p16 mayCDKN2 –13, inclCDKN2 . This loCDKN2 , 16. FurCDKN2 , 18, incCDKN2 .Examination of p16 immunohistochemical staining in CBN and in particular atypical CBN, which are difficult to differentiate from melanoma arising in or mimicking a CBN, has not yet been reported. In this study, we analyzed p16 expression in benign, mildly, moderately, and severely atypical CBN and compared these findings with cases of melanoma, malignant CBN, conventional benign, and atypical melanocytic nevi. We also examined Ki-67 in these lesions in order to compare this well-known marker to p16 to further evaluate the usefulness of p16 as an adjunct in distinguishing benign and atypical CBN from melanoma.Paraffin-embedded, formalin-fixed blocks were obtained from 15 CBN, 8 conventional and atypical melanocytic nevi, 12 melanomas, and 4 malignant CBN from the Kaiser Permanente Southern California pathology files. This study was approved by the Kaiser Permanente Southern California institutional review board. Among the CBN, one was conventional CBN, five were mildly atypical, four were moderately atypical, and five were severely atypical. The histologic grading was based on architectural features  and cytologic features . The conventional benign and atypical melanocytic nevi consisted of compound congenital nevi and atypical nevi ranging in atypia from mild to severe. The melanomas included a melanoma in situ, seven superficial spreading melanomas, two spitzoid melanomas, a nevoid melanoma, and a melanoma metastasis. In addition, we examined four frankly malignant cellular blue nevi (three primary and one recurrent). The depth of the invasive melanomas ranged from 0.3 mm to 5.0 mm.μm sections of the paraffin-embedded tissues. p16 staining was performed on all the specimens, Ki-67 was performed on all but one severely atypical cellular blue nevus, six of eight conventional nevi, and seven of 12 melanomas. HMB-45 was performed on three moderately/severely atypical CBN, four of eight conventional nevi, and six of 12 melanomas. p16 immunohistochemical staining was performed using the CINtec Histology Kit  which utilizes the p16 antibody clone E6H4.Immunohistochemical staining for p16, HMB-45 (only select cases), and Ki-67 was performed on 5 p16 (nuclear and cytoplasmic) and HMB-45 (cytoplasmic) staining intensity were graded on a scale of 0–3 , and the percentage of melanocytes positive for p16, HMB-45, and Ki-67 was determined. Each tumor was independently evaluated by two reviewers, and the results were averaged.t-test. ANOVA testing showed that p16 and Ki-67 staining results were statistically significant for the comparisons made among the multiple tumor categories.Statistical significance was calculated using the analysis of variance (ANOVA) test and the two-tailed, two sample unequal variance P = 0.02) and percentage of melanocytes positive with p16  in the CBN  and percentage of positive melanocytes staining with p16  , but not in percentage of melanocytes stained for p16   , Table 2 = 0.02) , Table 4P = 0.0002; N = 6) and moderately atypical , but not in CBN which were severely atypical   . Similar; N = 5) , as well; N = 5) . When di; N = 5) , but not; N = 5) . Percent; N = 5) , Table 2P = 0.009) and the percentage of melanocytes stained with p16  . This inverse relationship between Ki-67 and p16 corresponds to studies that have shown that proliferation rate as detected by Ki-67 has been associated with absent or minimal p16 staining in melanomas , 26.Few studies have compared p16 staining in atypical nevi versus benign nevi. Sanki et al.  showed tP values for percentage of melanocytes stained for p16 when CBN were compared to melanoma and malignant CBN. Unfortunately, p16 appears to not be useful in distinguishing severely atypical CBN from malignant CBN and melanoma. As the distinction between severely atypical CBN and malignant CBN is a very difficult diagnostic challenge, these findings are disappointing, but perhaps not surprising, as they likely reflect the overlapping histologic and biologic features between these tumors.In conclusion, p16 immunohistochemistry can be helpful to confirm the benign nature of a CBN when the staining is elevated, as seen in conventional, mildly, and moderately atypical CBN. As there are some cases of melanomas which do not lose p16 staining, this marker should not be used singly, but in conjunction with the histologic findings (which remain the gold standard) and other markers such as Ki-67 in making the distinction between atypical cellular blue nevi and melanoma. In interpreting p16 staining, the percentage of melanocytes stained for p16 should be considered more strongly than p16 staining intensity, as our results demonstrated lower


H1/TH17-polarized CD4+ T cells, B cells, and a plethora of myeloid cells, resulting in severe demyelination ultimately leading to a degeneration of neuronal structures. These pathological processes are substantially modulated by microglia, the resident immune competent cells of the CNS. In this overview, we summarize the current knowledge regarding the highly diverse and complex function of microglia during CNS autoimmunity in either promoting tissue injury or tissue repair. Hence, understanding microglia involvement in MS offers new exciting paths for therapeutic intervention.Multiple sclerosis (MS) is the most common autoimmune disease of the central nervous system (CNS) in the Western world. The disease is characterized histologically by the infiltration of encephalitogenic T Profound insights into the pathogenic mechanisms involved in MS as well as possible therapeutic interventions were gained through the use of experimental autoimmune encephalitis (EAE), the most used animal model for CNS autoimmunity [35-55 peptide is thought to be a monophasic chronically active disease without significant recovery and relapse phases, thereby only partially reflecting the clinical course found in MS patients [An autoimmune disease is characterized by the loss of self-tolerance of the immune system, which can be caused by either genetic or environmental factors or a combination of both . As a copse rate –5. Unforimmunity , 7. Seveimmunity , 9. Someimmunity . Howeverpatients . Neverth−-erythromyeloid precursors as the genuine microglia progenitors on a stem cell level [Microglia cells are resident tissue macrophages located in the CNS and are considered to be a patrolling immune competent cell type within the parenchyma –17. Micrll level . These pll level , 20. By ll level . In factll level , 23–25. in vivo imaging has revealed that microglia actively scavenge and monitor with their ramified branches the environment of the CNS for pathogens [γ and lipopolysaccharide (LPS) polarize microglia towards the M1 state and induce the release or expression of interleukin- (IL-) 1, IL-6, IL-12, IL-23, and inducible nitric oxide synthase (iNOS). On the other hand, the presence of IL-4, IL-10, and IL-13 turns microglia into M2 cells, which produce IL-10 and express arginase 1 [Due to their morphology, microglia cells are ascribed to be in a “resting” state under healthy conditions. This term is somewhat misleading, since athogens , 27. Upoathogens , 29. Theathogens , 30  or minocycline, which also attenuated EAE symptoms [H1 and TH17), while M2 microglia are designated as being protective in MS [For several decades, the activation of microglia has been described in the damaged CNS during the pathology of MS/EAE, reflecting an initial event in MS pathology . Even inuced EAE . In ordeenerated . In thissymptoms , 40. Recsymptoms , in micrsymptoms . Under psymptoms . The presymptoms . During symptoms . Thus, mve in MS , 44. Theα and thereby decreases the expression of C/EBP-α and indirectly Pu.1, both important myeloid regulatory transcription factors responsible for expression of activation markers [Recently, miRNA124 (miR-124) was identified to be a key regulator and modulator of microglia/monocyte activation . miR-124 markers . Finallyfl/fl mice, resulted in an early onset and more severe disease course, with an overly activated STAT3/4 signalling [In addition, suppressor of cytokine signalling (SOCS-) 3 was identified as another key player of microglia activation . SOCS prgnalling . MyeloidCytokines and chemokines are small secreted signalling molecules important for inter- and intracellular communication during inflammation. While both are mostly expressed at very low, basal levels in healthy conditions, the expression and secretion, primarily by microglia but also by astrocytes, are markedly increased upon CNS insult –50. Micrγ, tumour necrosis factor- (TNF-) α, IL-1β, and IL-6 are potent proinflammatory cytokines activating microglia and shifting them towards the cytotoxic M1 phenotype, thereby potentiating the inflammatory response [α, IL-1β, IL-2, IL-6, and IL-12, as well as iNOS and cyclooxygenase-2 (COX-2), binding of more than one cytokine is necessary [κB, JNK, ERK1/2, and p38 [κB signalling cascade in microglia/macrophages in LysMCreIκBαfl/fl mice worsens clinical symptoms of EAE and increases the expression of, for example, IL-1β and IL-6 [κB signalling is not only important in microglia, but also plays an important role in neuroectodermal-derived astrocytes during autoimmune inflammation of the CNS [ IFN-response –52  cells is induced [The roles of the proinflammatory cytokines IL-12, IL-23, and IL-17 produced by classically activated M1 microglia are more sophisticated –60 Figu. Interes17 cells , 60, 61.γ, TNF-α, and IL-1β, can induce the expression of CCL2 in the brain [hi mononuclear leukocytes into the CNS [ In addition to cytokines, microglia are also activated by and are able to release diverse chemoattractant chemokines, such as CCL2, CCL3, CCL4, CCL5, CXCL10, and/or CCL12 , 62, 63  the CNS , 66. Con the CNS –70. In a the CNS . In summ the CNS .α, or Fcγ receptor [Phagocytosis can be seen as a double-edged sword; on the one hand, it is beneficial by clearing cellular debris, but on other the hand it can also be destructive by inducing an oxidative burst . Microglreceptor . αPDGFR+ oligodendrocyte precursor recruitment [Interestingly, remaining myelin debris inhibits the regenerative remyelination, indicated by a loss of ruitment , 77. A druitment . Additioruitment . Thus, eH17, TH1) or anti-inflammatory T cells (TH2 or Treg) takes place, depending on the cytokine environment [MS/EAE pathology is characterized by harmful T cell infiltration . For T cironment . κB and p38 signalling [in vitro and reduced EAE severity in vivo [in vitro analyses revealed an inhibition of NF-κB signalling by compound A in microglia [β-Lapachone treatment, a natural substance from the bark of the lapacho tree, significantly reduced the expression of IL-12 family cytokines in microglia and thereby suppressed clinical severity [With the increased understanding of microglia function in MS/EAE pathology gained thus far, it might be possible to find and establish efficient therapies by targeting microglial responses. Since M1 microglia seem to be harmful during CNS autoimmune inflammation, most therapeutic ideas aim to either deactivate M1 microglia or turn them into M2 cells. Recently, galectin-  1 was identified as a potent microglia regulator . Gal-1 pgnalling . Finallygnalling . Similargnalling , 89. Tre in vivo , 89. In icroglia . β-Lapacseverity . β with synthetic ligands efficiently turned down microglia activation and thereby reduced EAE symptoms [ During the termination of the acute inflammatory reaction, microglia become deactivated . Therefosymptoms , 93. Recently, the use of reprogrammed embryonic stem (ES) cell-derived microglia-like cells (ESdM) for MS was suggested . IntraveActivation of microglia is a hallmark of MS/EAE pathology. During recent years more and more knowledge about how and why microglia become activated has accumulated. Subsequently, several investigations have already proven the feasibility of beneficially modulating microglia function in animal models of MS. Nevertheless, this path has to be taken further. There is a need for comprehensive and detailed analysis to further illuminate the triggering and signalling mechanisms of microglia in inflammation. Thoroughly understanding microglia action in the context of a disease will help us identify new targets for therapeutic approaches, which may ultimately be translated to the clinics.


Intramuscular fat (IMF) is one of the important factors influencing meat quality, however, for chickens, the molecular regulatory mechanisms underlying this trait have not yet been determined. In this study, a systematic identification of candidate genes and new pathways related to IMF deposition in chicken breast tissue has been made using gene expression profiles of two distinct breeds: Beijing-you (BJY), a slow-growing Chinese breed possessing high meat quality and Arbor Acres (AA), a commercial fast-growing broiler line.Agilent cDNA microarray analyses were conducted to determine gene expression profiles of breast muscle sampled at different developmental stages of BJY and AA chickens. Relative to d 1 when there is no detectable IMF, breast muscle at d 21, d 42, d 90 and d 120 (only for BJY) contained 1310 differentially expressed genes (DEGs) in BJY and 1080 DEGs in AA. Of these, 34–70 DEGs related to lipid metabolism or muscle development processes were examined further in each breed based on Gene Ontology (GO) analysis. The expression of several DEGs was correlated, positively or negatively, with the changing patterns of lipid content or breast weight across the ages sampled, indicating that those genes may play key roles in these developmental processes. In addition, based on KEGG pathway analysis of DEGs in both BJY and AA chickens, it was found that in addition to pathways affecting lipid metabolism , cell junction-related pathways , which play a prominent role in maintaining the integrity of tissues, could contribute to the IMF deposition.The results of this study identified potential candidate genes associated with chicken IMF deposition and imply that IMF deposition in chicken breast muscle is regulated and mediated not only by genes and pathways related to lipid metabolism and muscle development, but also by others involved in cell junctions. These findings establish the groundwork and provide new clues for deciphering the molecular mechanisms underlying IMF deposition in poultry. Further studies at the translational and posttranslational level are now required to validate the genes and pathways identified here. During the past decades, meat poultry breeding has been predominantly focused on accelerating growth rate and yields of breast and thigh meat. The impressive progress made in these traits, however, has been accompanied by deterioration of taste quality of the broiler meat and, in some markets, decreased acceptability of the meat by consumers ,2.Intramuscular fat (IMF), located in most species in the epimysium, perimysium and endomysium, is an important determinant of meat quality. A certain amount of IMF can enhance meat quality traits such as the flavor, juiciness, water holding capacity and tenderness -7. AdditCompared to beef, chicken meat does not present a marbled aspect and intramuscular fat is not visible and not anatomically separable; a higher proportion of IMF is represented by polar lipids, presumably within membranes. Previous studies found that the IMF content of chicken meat increased with age from d 28 to d 90  and may de novo synthesis of fatty acids occurs in adipose tissue [For livestock IMF, studies of the molecular mechanisms underlying IMF deposition have revealed large numbers of DEGs and signaling pathways including ADAMST4-signaling and insulin-signaling pathways -17. Compe tissue , chickene tissue -21. Untie tissue , liver [e tissue  or muscle tissue  have beeTo identify potential candidate genes affecting chicken IMF deposition, gene expression profiles in breast muscle of both Beijing-you  and Arbor Acres  chickens at different developmental stages were examined using Agilent cDNA microarray technology. Divergence of breast growth rates in BJY and AA chickens are shown in Table Based on the known DE genes, GO analysis was performed in each breed, and the enriched GO-terms (P < 0.05) in the ontology classification “Biological Process” were selected and are presented in Additional file To validate the microarrays, normal distribution analysis was performed with the results of the nine microarrays, and all of their coefficients were <0.5. Based on the 595 known genes that were shared as DEGs in both breeds, cluster analysis of all microarrays was performed Figure  using thTo further validate the results of microarray testing, q-PCR was used to examine the relative expression of 9 DE key genes selected in each breed at different ages. The results showed acceptable consistency between the results of q-PCR and the fold-change of DE genes in the microarrays Table .FGF4, MYH4, MYBPC1, MYH7B, MYL2, MYL10, MYL3, MYO15A, TGFA and WNT4  and genes related to lipid metabolism  were found  and phospholipids (PLIP) in breast tissue of the two breeds at different ages are provided in Table GDF3 and CAPNS1 in AA, and ANKRD1 and PLG in BJY were positively correlated (p < 0.05) with muscle development. The expression of BMP2K and MYBPC1 in AA, and CENPF, ELN, FGF7, FGFR1, MYBL2, MYCN and MYBPC1 in BJY were negatively correlated (p < 0.05) with that of muscle development  with PLIP contents while NACA, RBP7 and GLTPD1 in AA chickens, and LOC416618, ETFDH and GLTPD1 in BJY chickens were negatively correlated (p < 0.05) with PLIP contents. In the case of TG deposition, HMGCLL1, THBS1, UCP3 and SNX4 in AA, and EHHADH and SNX4 in BJY were positively correlated (p < 0.05) with TG contents and SGPL1, SH3PXD2B and THRSP in just the AA chickens were negatively correlated (p < 0.05) with contents of TG or IMF (Table MYBPC1), common to both breeds, was positively correlated with breast tissue weight across ages while 3 DEGs , common to the two breeds, were positively or negatively correlated with IMF, TG or PLIP contents  were examined. The expression of nt Table . For lipMYBPC1, GLTPD1, CETP, SNX4 transcripts in BJY chickens and in AA chickens were further measured by q-PCR. Consistent correlation between mRNA expression of these genes and muscle development and lipid deposition  were found and are shown in Figure The relative abundances of The regulation of IMF is possibly a function of complex pathway interactions involving muscle, fat and connective tissue , so examFat deposition in chickens takes place mainly in visceral adipose tissue and in muscle. The latter, IMF, is very important for sensory aspects of meat quality and there is increasing interest in improving quality, perhaps using marker-assisted selection for IMF. Although global gene expression surveys have been performed on visceral tissues , this st2 = 0.85) and AA (r2 = 0.72) chickens.Chicken microarrays were employed, each using pooled RNA samples . Such a pooling strategy can dramatically improve accuracy when only one array is available in each biological condition . PotentiCETPGLTPD1 and SNX4) and showed consistent trends with the changes in IMF, TG or PLIP contents across the ages examined  changed consistently with the changing patterns of breast tissue weight (Table MYBPC1 gene encodes a slow form(s) of MyBP C found in skeletal muscle and functions in the assembly and stabilization of sarcomeric M- and A-bands and regulates the contractile properties of actomyosin filaments. In chickens, increased levels of MyBP-C slow are found in dystrophic skeletal muscles [For key candidate genes affecting muscle development, one gene common to the two breeds , ACSL4, CD36, PLTP[adipocyte FABP and heart-type FABP genes and SNP markers from these genes were found to be associated with IMF content in Chinese chickens [FGF1, FGF4, FGF7, FGF16, FGFR1 and FGFR2) belonging to the FGF family and receptors were reflected in the MAPK signaling pathway.GO-term analysis was used to explore the function of DEGs and KEGG pathway analysis was used to explore the regulatory network underlying chicken IMF deposition. As expected, several well-known pathways related to lipid metabolism were found, including the MAPK and PPAR signaling pathways . Large nD36, PLTP-33. Thischickens ,35. SeveMYBPC1, MYBPC2, MYH4, MYH7BMYL10, MYL2, MYL3, MYO15A and MY01F) related to the biosynthesis of myosin, of which MYL10 and MYL2 encode proteins involved in the formation of tight junctions, focal adhesions and regulation of the actin cytoskeleton. Additionally, the CTNNB1 gene encodes β-catenin, one of the proteins constituting adherens junctions, and anchoring the actin cytoskeleton [ACTB encodes beta-actin, one of the non-muscle cytoskeletal actins playing a central role in shape determination, cytokinesis, and cell motility, along with cell-cell and cell-matrix interactions [FGF1, FGF4, FGF7, FGF16, FGFR1 and FGFR2) were reflected in the regulation of the actin cytoskeleton and may modulate morphogenetic processes involving cellular rearrangements and tissue remodeling [THBS1 and CD36 are involved in extracellular matrix (ECM)-receptor interaction [Of special interest, these GO and KEGG analyses provide the first demonstration that a series of pathways related to cell junctions might contribute to the deposition of IMF. DEGs related to muscle development included well-known genes  may play key roles in IMF developmental processes because their expressions were correlated with the changing patterns of lipid content or breast weight across the ages sampled in both two breeds. In addition, the results of KEGG pathway analysis imply that IMF deposition in chickens is regulated and mediated not only by genes and pathways related to lipid metabolism and muscle development, but also by others involved in cell junctions with the function in maintaining the integrity of tissues and signal transduction. These findings establish the groundwork and provide new clues for deciphering the molecular mechanisms underlying IMF deposition in poultry. Additional studies of translational and posttranslational effects will be required to complement these mRNA expression analyses.With aim of identifing global candidate genes and new pathways related to IMF deposition in chicken breast, Agilent cDNA microarray analyses were performed with both fast- and slow- growing breeds. Gene expression profiles of breast muscle sampled at different developmental stages of BJY and AA chickens were determined. Relative to d 1, breast muscle at d 21, d 42, d 90 and d 120 (only for BJY) contained 1310 DEGs in BJY and 1080 DEGs in AA. Several DEGs  and Arbor Acres , were performed in accordance with the Guidelines for Experimental Animals established by the Ministry of Science and Technology . Individuals within each breed had the same genetic background. Birds (60 BJY and 48 AA) were reared in stair-step caging under continuous lighting using standard conditions of temperature, humidity and ventilation. Chickens used for sample collection at d 1 were not fed. The same diet was fed to all chickens and was formulated to be intermediate between recommendations for the two breeds ,50. The pectoralis major muscle were excised, snap-frozen in liquid nitrogen and stored at −80°C. The entire right breast was collected and stored at −20°C for trait measurements.At each sampling age, d 1, d 21, d 42, d 90, and d 120 (only for BJY), six birds of similar weight from each breed were sacrificed for tissue collection. Samples of the left Intramuscular fat (IMF) content of breast muscle was determined by extraction with petroleum ether in a Soxhlet apparatus ,52 and epectoralis major muscle were homogenized using the method of Folch [Samples of the right of Folch . The conof Folch ,55 and Pof Folch ,57 kits Pectoralis major collected at each sampling age were extracted, and pooled within days and breeds for testing with microarrays. Microarray hybridization was carried out by GeneTech Biotechnology Limited Company  using Agilent Chicken Gene Chips (ID: 015068) with 42034 probes. Array scanning and data extraction were carried out following the standard protocol.Total RNA was isolated from breast muscle samples using Trizol reagent  according to the manufacturer's instructions and dissolved in RNase-free water at a final 2.0 μg/μl concentration. RNA from The normal distribution of signal plots in every chip was determined. Clustering was performed based on the DE genes in each chicken breed, using un-centered Pearson correlations and average linkage cluster 3.0, and was displayed in TreeView. Normalized fluorescence intensity values of each dye-swapped experiment were averaged separately for sample and reference channels. Thereafter, for each probe, averaged sample and reference fluorescence values were log2-transformed. Average linkage hierarchical clustering was performed using the Euclidian metric. In heat-maps, the color of features (probes) was determined by log2 (reference/sample).The distribution of expressed genes was analyzed by JMP4.0 according to their expression level. If the flag of a gene was “A” by the scanner according to the data normalization and results of Agilent Microarray Suite 4.0 software, it was considered to be "not detected", and hence "not expressed" in this study. Similarly, the genes with “P” flags were considered to be “expressed transcripts”. Expressed transcripts were defined as being present in samples of at least one sampled age and were used for all following studies. The expression value of each probe set was normalized and calibrated using the RMA method.Screening of differentially expressed genes (DEGs) was performed on the basis of differences in the IMF, TG and PLIP contents at different ages in each breed. Expression at d 1 was used as the controls, and comparisons were made within each breed at d 21, 42, 90 and 120 (only for BJY). Genes were considered to be DEGs only when the fold-change in abundance for all comparisons exceeded 2.0.To avoid amplification of any residual genomic DNA, all PCR primers were placed at or just outside the exon/exon junctions and specificity was determined with BLASTN . Melting curves were constructed to verify that only a single PCR product was amplified. Within runs, samples were assayed in triplicate with standard deviations of threshold cycle (CT) values not exceeding 0.5, and each q-PCR run was repeated at least twice. Negative (without template) reactions were performed within each assay.Quantification of the transcripts was determined using standard curves with 10-fold serial dilutions of cDNA  with breast tissue weight, or content of IMF, TG or PLIP, this gene was considered to be a key DE gene.KEGG pathway -60 inforACSF3, Acyl-CoA synthetase family member 3; ACSL1, Acyl-CoA synthetase long-chain family member 1; ACSS(1&2), Acyl-CoA synthetase short-chain family member; ACTB, Actin, beta; ADAMST4, A disintegrin and metalloproteinase with thrombospondin motif 4; ANKRD1, Ankyrin repeat domain 1; APOH, Apolipoprotein H; BMP, Bone morphogenetic protein; BMP2K, BMP-2-inducible protein kinase; CAMs, Cell adhesion molecules; CAPNS1, calpain, small subunit 1; CD36, Cluster of Differentiation 36; CD44, CD44-like protein; CENPF, Centromere protein F; CETP, Cholesteryl ester transfer protein; CH25H, Cholesterol 25-hydroxylase; CTNNB1, Catenin (cadherin-associated protein), beta 1; CYP17A1, Cytochrome P450, family 17, subfamily A, polypeptide 1; DCK, Deoxycytidine kinase; DGAT2, Diacylglycerol O-acyltransferase homolog 2; ECM, Extracellular matrix; EGF, Epidermal growth factor; EHHADH, Enoyl-Coenzyme A, hydratase/3-hydroxyacyl Coenzyme A dehydrogenase; ELN, Elastin; ErbB, Erythroblastic Leukemia Viral Ongene Homolog; ETFDH, Electron-transferring-flavoprotein dehydrogenase; FABP, Fatty acid binding protein; FGF, Fibroblast growth factor; FGFR, Fibroblast growth factor receptor; GDF3, Growth differentiation factor 3; GLTPD1, Glycolipid transfer protein domain containing 1; HMGCLL1, 3-hydroxymethyl-3-methylglutaryl-Coenzyme A lyase-like 1; KEGG, Kyoto Encyclopedia of Genes and Genomes; LDLR, Low density lipoprotein receptor; LOC416618, NAD(P) dependent steroid dehydrogenase-like,; MAP2K3, Mitogen-activated protein kinase kinase 3; MAPK, Mitogen-activated protein kinase; MBOAT2, Membrane bound O-acyltransferase domain containing 2; MYBL2, V-myb myeloblastosis viral oncogene homolog (avian)-like 2; MYBPC1, Myosin binding protein C, slow type; MYCN, V-myc myelocytomatosis viral related oncogene, neuroblastoma derived; MYF5, Myogenic factor 5; MYH, Myosin, heavy polypeptide; MYL, Myosin, light chain; MYO15A, Myosin XVA; NACA, Nascent polypeptide-associated complex alpha subunit; NR3C2, Nuclear receptor subfamily 3, group C, member 2; PGK1, Phosphoglycerate kinase 1; PLTP, Phospholipid transfer protein; PLG, Plasminogen; PPAR, Peroxisome proliferator-activated receptor; PPARGC1A, Peroxisome proliferator-activated receptor gamma, coactivator 1 alpha; RBP7, Retinol binding protein 7; RCJMB04_13o20, NSFL1 (p97) cofactor (p47); RXRA, Retinoid X receptor-alpha; SGPL1, Sphingosine-1-phosphate lyase 1; SH3PXD2B, SH3 and PX domains 2B; SNX, Sorting nexin; TGFA, Transforming growth factor, alpha; THBS1, Thrombospondin-1; THRSP, Thyroid hormone responsive, SPOT14 homolog; UCP3, Uncoupling protein 3; YWHAH, Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein; WNT4, Wingless-type MMTV integration site family, member 4.Annotation and changing of 1746 common DE genes in AA and BJY chickens.Click here for fileCommon Enriched GO terms among the differentially expressed genes in both AA and BYJ chickens.Click here for fileKey genes related to lipid metabolism and muscle development in BJY chicken.Click here for fileKey genes related to lipid metabolism and muscle development in AA chicken.Click here for fileCommon DE genes related to muscle development or lipid metabolism in AA and BJY.Click here for fileIdentification of enriched KEGG pathways based on known DEGs related to muscle development and lipid metabolism in AA and BJY chickens.Click here for fileSelected qPCR primer sequences and accession numbers.Click here for file


Glomerular diseases and obstructive uropathies are the two most frequent causes of chronic kidney disease (CKD) in children. Recently, biomarkers have become a focus of clinical research as potentially useful diagnostic tools in pediatric renal diseases. Among several putative biomarkers, chemokines emerge as promising molecules since they play relevant roles in the pathophysiology of pediatric renal diseases. The evaluation of these inflammatory mediators might help in the management of diverse renal diseases in children and the detection of patients at high risk to develop CKD. The aim of this paper is to revise general aspects of chemokines and the potential link between chemokines and the most common pediatric renal diseases by including experimental and clinical evidence. Chemokines constitute a large family of low molecular-weight cytokines whose main action is the recruitment of leukocyte subsets under homeostatic conditions and inflammatio; the word “chemokine” is the contraction of the terms “chemotactic” and “cytokine” . More spTo date, approximately 50 chemokines and 20 receptors have been described in humans , 3, 4. Cα , CX3CL1/fractalkine, and CXCL8/IL8 (interleukin 8) [With respect to renal diseases, there is much evidence that leukocyte infiltration is mediated by inflammatory chemokines released by various cell types , 5. Infieukin 8) . Messengeukin 8) .There are several techniques to measure chemokine—protein or mRNA—in tissues and body fluids. For example, chemokines could be directly measured in renal tissue by immunohistochemical or immunofluorescent techniques or by evaluating their levels in supernatants of homogenized tissues. In patients with renal diseases, the direct exam of tissue samples would be ideal, since it may evaluate the affected organ. However, kidney biopsy is an aggressive procedure and could be harmful. On the other hand, enzyme-linked immunosorbent assay (ELISA) or flow cytometry based techniques are less invasive and more useful for clinical purposes by measuring the levels of chemokines in urine or blood samples –12. AlteA number of studies have shown the relation between renal diseases and chemokines production –6. IndeeCXCL8/IL-8 is a chemokine responsible for neutrophil infiltration into the urinary tract with an important role in acute pyelonephritis , 16. In CCL2/MCP-1 was the first CC chemokine to be discovered, acting as a potent chemotactic factor for monocytes, and has been one of the most studied biomarkers , 17–28. CCL5/RANTES is a chemokine produced by human T lymphocytes at a “late” stage (3–5 days) after activation through their T-cell receptors. It is broadly chemoattractive for T lymphocytes, monocytes, natural killer cells, basophils, and eosinophils and can also activate immune cells. This chemokine is involved in AIDS, cancer, atherosclerosis, asthma, organ transplantation, and autoimmune diseases such as arthritis, diabetes, and glomerulonephritis .α, IL-1β, G-CSF, and GM-CSF. IL-17 cytokines mediate the chemotaxis of neutrophils to sites of infection and upregulate the intercellular adhesion molecule-1 [In addition, some cytokines clearly interact with chemokines. As an example, Th-17 cells are a subset of Th cells, which produce IL-17A and IL-17F, members of the IL-17 cytokine family. These cytokines induce various cytokines/chemokines including CXCL8/IL8, IL-6, CCL2/MCP-1, TNF-lecule-1 . IL-17A lecule-1 –34.In the last five years some, clinical studies measuring chemokines have been done in many forms of glomerular diseases such as IgA nephropathy, lupus nephritis, and idiopathic nephrotic syndrome (INS) including minimal change lesion (MCNS) and focal segmental glomerulosclerosis (FSGS).β, CCL2/MCP-1, IL-17, IFN-γ, IL-6, and IL-10) have been evaluated in IgA patients and some of them seemed to predict the outcome of the disease [IgA nephropathy is diagnosed by the predominance of IgA deposits in the glomerular mesangium and is present in around 13.8% of renal biopsies of children, being the second more frequent renal disease, and the mesangioproliferative glomerulonephritis is the most common histological presentation . Several disease . As an e disease .α, CCL5/RANTES, VEGF, IL-6, CXCL8/IL-8, IL-17, CCL2/MCP-1, CXCL3/MIP-1β, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IFN-γ, and TNF-α) were tested for correlation with lupus activity by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI-1) [α levels were significantly increased in patients with nephritis when compared to first-degree relatives and healthy controls, and TNF-α levels were significantly increased in patients with active disease [α was elevated in 12 patients with active lupus nephritis compared with inactive SLE and healthy subjects and declined after clinical remission [Systemic lupus erythematosus (SLE) is a multisystemic autoimmune disease affecting predominantly women and 2/3 of cases occur in the first two decades of age. Lupus nephritis is a very common feature observed in 50–67% of children with SLE . A wide LEDAI-1) . The stuLEDAI-1) . In a st disease . Serum semission . IL-17 aemission , 44.β might differentiate between FSGS and MCNS, but it seemed not to be a biomarker of steroid responsiveness [MCNS is the most common cause of nephrotic syndrome in children. The authors have proposed that this disease reflects a disorder of T-lymphocytes and some patients can progress to FSGS . Araya asiveness . Our ressiveness . We founsiveness . These fsiveness . More resiveness . In addisiveness . This stsiveness . Urinarysiveness .spectrum of malformations that occur at the level of the kidney , collecting system , bladder , or urethra  [Congenital anomalies of the kidney and urinary tract (CAKUT) comprise a  valves) . A varie valves) . An alte valves) , 50.γ-interferon-inducible protein (CXCL10/IP-10) have been evaluated in experimental hydronephrosis [In this regard, chemokines like CCL2/MCP-1, CCL5/RANTES, macrophage inflammatory protein-2 (CXCL2/MIP-2), and ephrosis –53. CCL2ephrosis , 54. Steephrosis . These aephrosis . In addiephrosis . An inteephrosis . Crismanephrosis . At 7 daephrosis . The stuephrosis .It should also be pointed that very few data about the role of chemokines in CAKUT were provided by clinical studies and the majority of them evaluated ureteropelvic junction obstruction (UPJO) and vesicoureteral reflux (VUR).UPJO is the most common cause of severe hydronephrosis in children –61. UPJOPostnatal differentiation between obstructive and nonobstructive hydronephrosis is quite difficult. Several studies have been made in patients with UPJO in order to find out noninvasive biomarkers to allow the diagnosis and treatment of these patients , 63, 64.α, CXCL10/IP-10, and CCL5/RANTES were increased in urine from the obstructed kidney compared to urine from the contralateral nonobstructed kidney [Healthy children presented high expression of EGF mRNA in renal tissue, whereas CCL2/MCP-1 mRNA was normally undetectable. On the other hand, in UPJO patients, CCL2/MCP-1 gene expression was strikingly increased at the tubulointerstitial level, while the EGF gene expression was markedly reduced. The interstitial mononuclear cell infiltrate in UPJO patients was strictly correlated with the degree of tubulointerstitial damage , 66. Accd kidney . These ud kidney .Taranta-Janusz and coworkers compared obstructed prenatal hydronephrosis cases (who underwent surgery) with nonsurgically managed cases and with healthy subjects (control group) . These aVUR is a congenital anomaly that increases the risk of repeated pyelonephritis and, consequently, can result in renal scarring, renin-mediated hypertension, and, in some cases, renal insufficiency , 71. VURμmol, the sensitivity of this marker in diagnosing VUR was 88%, the specificity 69%, the positive prognostic value 66%, and the negative prognostic value 89%. At higher cutoff concentrations, specificity of the marker increased, but sensitivity rapidly decreased [The inflammatory process in VUR is ongoing despite the occurrence or not of urinary tract infection (UTI). The elevated urinary level of CXCL8/IL-8 in children with reflux and without UTI might contribute to reflux nephropathy –75. Haraecreased . Our resecreased .New diagnostic approaches to and alternative therapies for pediatric renal diseases are clearly necessary. In this context, research into biomarkers has reached great importance. In 2001, an NIH working group standardized the definition of a biomarker as “a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention” . In thisWe believed that spot urine measurements of chemokines might become useful in pediatric renal diseases even at clinical practice. For instance, CCL2/MCP-1 and CXCL8/IL-8 are the chemokines more commonly associated with pediatric renal diseases , 73–75.In regard to glomerular diseases, high urinary levels of CCL2/MCP-1 have been frequently related to FSGS , 45, 46,In pediatric patients with CAKUT, the chemokine CCL2/MCP-1 has been associated with urinary tract obstruction in UPJO –68, wherYet, in spite of great advances in our knowledge about the pathophysiological mechanism linking the chemokines to renal diseases, much remains to be elucidated. Furthermore, the usefulness of chemokine measurements in clinical practice still needs to be established.


To the Editor: Toxic shock syndrome (TSS), which can be life threatening, is defined by clinical and laboratory evidence of fever, rash, desquamation, hypotension, and multiple organ failure caused by Staphylococcus aureus toxins. TSS caused by methicillin-resistant S. aureus (MRSA) strains has been found extensively in Japan (We report a case of TSS due to an MRSA strain that produced a TSS toxin 1 (TSST-1). A 54-year-old woman was admitted to the emergency ward of Brugmann University Hospital, Brussels, with a 2-day history of myalgia, diarrhea, and vomiting. She had undergone surgery for a palate neoplasia 2 months earlier, and again 2 weeks earlier, in another hospital. After the second operation, she had been treated for a local scar infection with amoxicillin–clavulanic acid for 1 week.3 with 97% polynuclear neutrophils, platelets 90,000/mm3, creatinine 2.1 mg/dL, bicarbonate 13 mEq/L, C-reactive protein 43.7 mg/dL, creatine kinase 514 U/L. Cultures of blood, stool, and urine samples were negative for microbial agents. Puncture of the wound released 12 mL of pus; culture of the pus sample yielded an MRSA strain harboring a TSST-1 gene, detected by multiplex polymerase chain reaction as previously described  type II. This clone belongs to the sequence type (ST) 5-SCCmec II clone, formerly named "New-York/Japan clone," which has been associated with neonatal TSS–like exanthematous disease in Japanese hospitals  (S. aureus, possibly MRSA. The possibility of MRSA must be considered when initiating antimicrobial agents to treat TSS.We immediately treated our patient with teicoplanin and clindamycin because we suspected a nosocomial infection with


Mitosis is tightly regulated and any errors in this process often lead to aneuploidy, genomic instability, and tumorigenesis. Deregulation of mitotic kinases is significantly associated with improper cell division and aneuploidy. Because of their importance during mitosis and the relevance to cancer, mitotic kinase signaling has been extensively studied over the past few decades and, as a result, several mitotic kinase inhibitors have been developed. Despite promising preclinical results, targeting mitotic kinases for cancer therapy faces numerous challenges, including safety and patient selection issues. Therefore, there is an urgent need to better understand the molecular mechanisms underlying mitotic kinase signaling and its interactive network. Increasing evidence suggests that tumor suppressor p53 functions at the center of the mitotic kinase signaling network. In response to mitotic spindle damage, multiple mitotic kinases phosphorylate p53 to either activate or deactivate p53-mediated signaling. p53 can also regulate the expression and function of mitotic kinases, suggesting the existence of a network of mutual regulation, which can be positive or negative, between mitotic kinases and p53 signaling. Therefore, deciphering this regulatory network will provide knowledge to overcome current limitations of targeting mitotic kinases and further improve the results of targeted therapy. Mitosis involves a highly orchestrated and fine-tuned sequence of events to properly transfer genetic information to the next generation by cell division , 2. It iMultiple phosphorylation and proteolysis events play important roles in the regulation of mitotic progression and cytokinesis , 2. Numep53 is one of the most frequently mutated or deleted genes in human cancers and plays a role in many cellular processes, including cell growth, differentiation, senescence, and DNA repair (reviewed in [p53 is located on chromosome 17 (17 p13) and consists of an N-terminal transactivation domain, a central specific DNA-binding domain and a C-terminal domain, containing a tetramerization domain and regulatory region [ Many studies have reported that deregulation of these mitotic kinases causes mitotic failure and aneuploidy and is closely associated with genomic instability and tumorigenesis , 36–38. iewed in ). In addiewed in , 43. Theiewed in ). The huy region . At leasy region  and impoy region , Thr-18 y region , and Sery region  are crity region . On the y region  and Thr-y region  often su Under normal circumstances, cells induce the p53-dependent transcriptional activation, cell cycle arrest, and apoptosis in response to mitotic defects or DNA damage . However In this paper, we will specifically focus on the classic mitotic kinases, including Aurora kinases, Plks, Bub1, Mps1, and BubR1, and their roles in regulating p53 protein stability and activity.Drosophila melanogaster mutants having defects in mitotic spindle-pole formation [Ipl1 gene is essential for maintaining genome stability through its roles in chromosome segregation, spindle checkpoint, mitotic spindle disassembly, and cytokinesis [Caenorhabditis elegans has two Aurora kinases, Aurora/Ipl1-related-1 and -2 (AIR-1 and AIR-2), and they are thought to be key regulators of mitotic spindle assembly and dynamics [Aurora kinases belong to a highly conserved family of serine/threonine kinases crucial for chromosome segregation, condensation, and spindle assembly . The firormation . Subsequormation . The Iplokinesis , 68. Caedynamics , 70. Thrdynamics , and fundynamics . Dereguldynamics . Not surdynamics –76. Sincin vitro and in vivo. Phosphorylation of Ser-215 but not Ser-315 inhibits p53 DNA binding and its transactivational activity [vice versa [ Xenopus p53 can block Xenopus Aurora A's ability to transform cells [ Aurora A is ubiquitously expressed in proliferating cells and its activity is tightly regulated through the cell cycle . Both thactivity , whereasactivity . These factivity . hnRNPK activity , suggestce versa . In addirm cells , further Aurora B is a member of the chromosome passenger complex (CPC), a key regulator of chromosome segregation, histone modification, and cytokinesis during mitosis , 94. The, indicating that Aurora C plays a critical role in meiosis. Recent studies show that Aurora B and C have similar structural and functional properties [ In contrast to Aurora A and B, the biological function of Aurora C has not been well-defined. Aurora C was first discovered in mouse sperm and eggs using a kinase screen . While Aoperties . Inhibitoperties . Aurora operties . Moreoveoperties , 106,   polo gene of Drosophila melanogaster, whose mutation causes a high frequency of abnormal mitosis and meiosis [Saccharomyces cerevisiae, [Schizosaccharomyces pombe [Caenorhabditis elegans [Xenopus laevis [Plks are a family of highly conserved serine/threonine protein kinases  named af meiosis . Subsequevisiae, , Plo1p ies pombe , Plc1, P elegans , 112, ans laevis –115. In s laevis –125. Alls laevis  and shars laevis , 126. It is now widely recognized that Plks are key regulators of mitosis, meiosis, and cytokinesis , 128 as in vitro and its transcriptional activity and proapoptotic function are inhibited by direct interaction and phosphorylation of Plk1 [ Plk1 reaches peak expression during G2/M phase and kinase activity during mitosis , 129. Pl of Plk1 . Plk1 ca of Plk1 . Plk1 ph of Plk1 . Topors  of Plk1 and funct of Plk1 , 149. Ph of Plk1 . G2 and  of Plk1 , 151 and of Plk1 , 152. Ph of Plk1 , 153. Pl of Plk1 , 154 and of Plk1 , 153. In of Plk1 , 63.Plk2 and Plk3 are serum-inducible immediate early response genes [Plk2 and Plk3 function as tumor suppressors in the p53-mediated signaling pathways to protect cell from DNA damage or oxidative stress . Activaiewed in . Plk2 apiewed in . Promoteid cells . Plk3 has shown deregulated expression of Plk3 in various types of cancers [ Plk3 plays an important role in the regulation of mitosis and DNA damage checkpoint , 162. It cancers , 163, su cancers  and colo cancers . Overexp cancers  and indu cancers . In resp cancers , 162. Pl cancers , 169 and Plk4 shares relatively little sequence homology with other members of Plks . Plk4 isPlk5 shows that the promoter region of Plk5 contains several p53 binding motifs; however, no such regulatory mechanisms have yet been found [Ras oncogene, suggesting that Plk5 may function as a tumor suppressor gene in brain cancer [A fifth member of the Plk family, Plk5, is mainly expressed in differentiated tissues, such as the brain, eye, and ovary , whereasen found . Interesn cancer , 181. Saccharomyces cerevisiae along with mitotic arrest-deficient 1, 2, and 3  in mammals), Bub3, and Mps1 [in vitro, possibly inducing cellular senescence [ Bub1 belongs to a small group of serine/threonine kinases that play multiple roles in chromosome segregation and spindle checkpoint during mitosis . Bub1 waand Mps1 , 184. Aland Mps1 , 184. Deand Mps1 –187. APCand Mps1 . Bub1 enand Mps1 . Phosphoand Mps1 –192. Buband Mps1 . It has and Mps1 . In addinescence . An intenescence . Schizosaccharomyces pombe [ Arabidopsis thaliana [ Xenopus laevis [p53 mutation, a basal-like phenotype of breast cancer and a poor prognosis outcome [Mps1 has an essential role in centrosome duplication, checkpoint signaling, cytokinesis, and development in organisms from yeast to mammalian –197. Kines pombe , PPK1 inthaliana , xMps1ins laevis  and mMpss laevis . Mps1 acs laevis , 202 ands laevis . Dereguls laevis , 204 ands laevis . The kins laevis , 206. Fos laevis . In addis laevis , at Ser-s laevis . Mps1-mes laevis ; thus, is laevis , 208. Ths laevis , 208. In outcome . These fBubR1 haploinsufficiency display a genetic instability phenotype due to underlying defects in DNA repair and chromosomal segregation [p53-null cells, inhibition of BubR1 expression enhances chromosomal instability and polyploidy; conversely, overexpression of BubR1 restores the checkpoint function, suppresses centrosome amplification, and selectively eliminates cells with amplified centrosomes [BubR1 in cancers are very rare [BubR1 is the mammalian homolog of yeast Mad3 and Bub1 , 210. Itregation . Moreoveregation . The redregation . Increasregation . BubR1 iregation . The expregation . In p53-trosomes . Interestrosomes .Despite ery rare .Thanks to advances in proteomics technology, many of the substrates for mitotic kinases have been identified, such as those listed above; however, the functional significance of these phosphorylation events has not been explored thoroughly. Therefore, dissecting the functional consequences of mitotic kinase-mediated phosphorylation should be given high priority to better understand their roles in mitosis. p53  or deregulation of mitotic kinases), p53 cannot be activated when damage occurs to the mitotic spindle, thereby inducing mitotic slippage and preventing apoptosis  . Therefo Studies have shown that depletion/inhibition of Aurora A, Aurora B, Plk1, or Bub1 induces cellular senescence or cell death in a p53-dependent or -independent but p73-dependent manner in many different cell types , 223–231


Annexin A2 (ANXA2) orchestrates multiple biologic processes and clinical associations, especially in cancer progression. The structure of ANXA2 affects its cellular localization and function. However, posttranslational modification and protease-mediated N-terminal cleavage also play critical roles in regulating ANXA2. ANXA2 expression levels vary among different types of cancers. With some cancers, ANXA2 can be used for the detection and diagnosis of cancer and for monitoring cancer progression. ANXA2 is also required for drug-resistance. This review discusses the feasibility of ANXA2 which is active in cancer development and can be a therapeutic target in cancer management. Annexin A2 , a 36 kDa protein , is locaκB nuclear translocation [β- and 12-O-tetradecanoylphorbol 13-acetate-induced tumor necrosis factor-α shedding by interacting with ADAM metallopeptidase domain 17. Inhibition of ANXA2 may be a therapeutic strategy for inflammation-associated disease progression [Intracellular ANXA2 has been reported to play roles in exocytosis , endocytlocation . Genotoxlocation . ANXA2, location . ANXA2 alocation . Knockdolocation . ANXA2 igression . Howevergression .Extracellular ANXA2 plays a role in phagocytosis , fibrinoα secretion through TLR4/MyD88- and TRIF-dependent pathways [As for the function of ANXA2 in immune regulation, recent studies show that secreted ANXA2 can activate macrophage for IL-1, IL-6, and TNF-pathways , 34. Thepathways  and hepapathways . Dendritpathways . Under Dpathways . It may pathways , 40.2+/phospholipid-stimulated kinases such as Ca2+-dependent protein kinases C (cPKCs), whereas one of the cPKC substrates, ANXAV, inhibits the phosphorylation of ANXA2 but other cPKC substrates have no effect on kinases that are only activated by phospholipids [The membrane association of ANXA2 can be induced by IL-6 in A549 cells via an unknown mechanism . Transloholipids . On the holipids . pp60 Srholipids . How is holipids . In addiholipids . Acetylaholipids . Abnormaholipids .2+ and requires an intact contractile apparatus [In vitro, cell cycle inhibition or apoptotic stimuli, such as treatment with chemotherapeutics, ER stress, or growth factor withdrawal, induce ANXA2 cleavage through a synergistic interaction of serine/threonine kinase glycogen synthase kinase-3 and serine protease Omi/HrA2. However, the overexpression of ANXA2 cannot reverse the cleavage [ANXA2 heterotetramerization is required for its extracellular function. N-terminal ANXA2 can stabilize intracellular p11 through direct binding and masking the autonomous p11 polyubiquitination signal, thus preventing proteasomal degradation . The N-tpparatus . In chappparatus . Howeverpparatus , 52. In cleavage , 54. Reccleavage  found thcleavage .Increased expression of ANXA2 is frequently observed in a broad spectrum of cancer cells. Overall, ANXA2 is overexpressed in acute lymphoblastic leukemia (ALL) , APL 5858, breasActually, compared with the peripheral-blood leukocytes or mononuclear cells from bone marrow from APL patients, the expression of ANXA2 in AML and ALL is lower . On the Compared to other leukemic cell types, leukemic cells from patients with APL or NB4 and HL60 cell lines display elevated levels of ANXA2 protein expression, as detected by a fluorescent-tagged antibody. The t-positive APL cells stimulated the generation of cell-surface, t-PA-dependent plasmin twice as efficiently as the t-negative cells. An abnormally high level of ANXA2 on APL cells increases the production of plasmin, a fibrinolytic protein. Thus, overexpression of ANXA2 may contribute to the hemorrhagic complications of APL . Howeverin situ. Furthermore, ANXA2 is selectively expressed in the invasive/metastatic MDA-MB231 cell but not in the noninvasive/nonmetastatic MCF-7. ANXA2-dependent, localized plasmin generation by human breast cancer cells could contribute to angiogenesis and metastasis [ANXA2 is undetectable in normal and hyperplastic ductal tissue samples. By contrast, ANXA2 is consistently expressed in invasive breast cancer and ductal carcinoma tastasis , 72. In tastasis  has showtastasis . A recentastasis . Furthertastasis .In 105 cases of primary colorectal carcinoma tissues, ANXA2 is overexpressed in the cancer cell membrane of the carcinoma cells more than in tumor stroma fibrous tissue, the muscularis propria, the vessel wall, and the adjacent normal bowel wall. Moreover, the expression of ANXA2 is positively correlated with histological type, tumor size, depth of invasion, and pathological tumor-node-metastasis (pTNM) stage . In 150 There is a clinical association between upregulated ANXA2 and tumorigenesis in gastric carcinoma. However, there is little understanding of the molecular function of ANXA2 in this context. Emoto et al. demonstrated that thirty-three percent of 153 primary gastric carcinoma patients are immunopositive for ANXA2; however, overexpression of ANXA2 is correlated with differentiation, lymph node metastasis, and venous invasion . It has in vivo, but lack of ANXA2 in stromal cells has no effect on glioma growth [ANXA2 protein is highly expressed in human high grade, but not in low grade, primary gliomas tissues. The expression of ANXA2 is highest in glioblastoma multiforme, intermediate in anaplastic astrocytomas, and lowest in astrocytomas. In contrast to the usual cytoplasmic localization of ANXA2, distinct nuclear staining has been found in many of the specimens . Knockdoa growth . The miga growth , 80. Fura growth .P < 0.01). Interestingly, both the tyrosine phosphorylation and protein expression of ANXA2 are elevated in HCC compared to normal or cirrhosis tissue [μL) and 78.4% (58/74) of alpha-fetoprotein (AFP)-negative  patients. Thus, ANXA2 may act as an independent serological candidate for the early detection of HCC, especially in early stage cases with normal serum AFP [ANXA2 is rarely detected in either normal or chronic hepatic tissues but is overexpressed transcriptionally and translationally in tumorous and nontumorous regions of HCC , especially in poorly differentiated HCC  . ANXA2 iMyeloma cells from MM patients display an increased expression of ANXA2 compared to plasma cells from normal subjects. In addition, ANXA2 expression in the MM cell lines U266 and RPMI8226 is elevated compared to other hematologic cell lines. Furthermore, ANXA2 has been shown to be required for the antiapoptosis, proliferation, and invasive potential of the MM cell lines U266 and RPMI8226 . Thus, AVishwanatha et al. found that in normal pancreas, ANXA2 expression is observed in ductal and ductular cells and there is no expression in acinar or islet cells. ANXA2 mRNA and protein levels in five established human pancreatic adenocarcinoma cell lines are elevated 5- to 15-fold compared with normal pancreas. However, higher expression of ANXA2 (2- to 8-fold) is observed in three primary pancreatic tumors and one metastatic tumor compared with normal pancreas . FurtherANXA2 is overexpressed in human B-cell lymphoma cell lines but is significantly reduced in Raji, OMA-BL-1 and REH cells compare with normal B-cells . Ohno etANXA2 expression is decreased in esophageal squamous cell carcinoma (ESCC) , 95, ostP < 0.05) and pathological differentiation (P < 0.05) [The mRNA and protein expression of ANXA2 is lower in only 22 ESCC tumor cells in the patient tissues than that in its paired normal counterpart , and thi < 0.05) . The abein vitro phenotypes that are often used to assess metastatic potential, including motility, adhesion, and proliferation. This study demonstrated, surprisingly, that ANXA2 is involved in the differentiation-induced mineralization process of osteoblastic cells in vitro and may decrease osteosarcoma aggressiveness [In osteosarcoma, the ANXA2 mRNA and protein levels are inversely correlated with metastatic potential in a subset of human osteosarcoma tumor specimens. However, overexpression of ANXA2 in SaOSLM2 cells did not alter several siveness .P < 0.01). However, in some high-grade tumors, ANXA2 facilitates the adhesion-dependent or -independent growth of prostate cancer through the MAPK pathway [ANXA2 heavy (p36) and light (p11) chains in 31 of 31 prostate cancer specimens are undetectable by immunohistochemistry, suggesting a loss of ANX2A expression in cancer cells, whereas basal cells are positively stained . ANXA2 e pathway  or inter pathway . Current pathway .Some of the earliest studies of ANXA2 demonstrated its expression on the APL surface and its role as a receptor for tPA and plasminogen following fibrinolysis . In pancIn addition to metastasis and invasion, previous studies have shown that ANXA2 is involved in tumor cell proliferation , 66, 103ANXA2 has been studied for many years, especially in tumors. Recently, the potential role of ANXA2 in immunology gives a hint on the interaction between immune cells and cancer cells in the microenvironment. In this paper, we summarize the expression and structure of ANXA2 as well as its function in cancer cells or primary cancer tissues. In the future, ANXA2 may be a potential prognostic factor, a potential diagnostic factor for cancer, and a therapeutic target for new drug development after analysis in large scale, well-controlled, prospective clinical trials.


Asthma is characterized by the association of airway hyperresponsiveness (AHR), inflammation, and remodelling. The aim of the present article is to review the pivotal role of airway smooth muscle (ASM) in the pathophysiology of asthma. ASM is the main effector of AHR. The mechanisms of AHR in asthma may involve a larger release of contractile mediators and/or a lower release of relaxant mediators, an improved ASM cell excitation/contraction coupling, and/or an alteration in the contraction/load coupling. Beyond its contractile function, ASM is also involved in bronchial inflammation and remodelling. Whereas ASM is a target of the inflammatory process, it can also display proinflammatory and immunomodulatory functions, through its synthetic properties and the expression of a wide range of cell surface molecules. ASM remodelling represents a key feature of asthmatic bronchial remodelling. ASM also plays a role in promoting complementary airway structural alterations, in particular by its synthetic function. The pathophysiology of asthma is characterized by the association of airway hyperresponsiveness (AHR), inflammation, and remodelling –3. AHR iα, or histamine, which, respectively, activate ASM cell membrane receptors TP, FP, and H1 [AHR is usually described as either nonspecific or specific AHR . Nonspec, and H1 . For ins, and H1 . Cell vo, and H1 . On the , and H1  and indu, and H1 .The mechanisms of AHR in asthma may involve (i) a larger release of contractile mediators, (ii) a lower release of relaxant mediators, (iii) an improved ASM cell excitation/contraction coupling, and/or (iv) an alteration in the contraction/load coupling.α, PGD2, and thromboxane TXA2. The concentration of these prostaglandins, and also that of histamine, is increased in asthma [The contraction of ASM can be induced by a variety of extracellular messengers, which act through interaction with specific receptors at the site of the plasma membrane of ASM . These cn asthma . Similarn asthma , and LTBn asthma . Such cyn asthma , as welln asthma , anothern asthma , producen asthma . The reln asthma , 40. It n asthma .β2-adrenergic receptor sometimes associated with its desensitization has been suggested in asthma following chronic medication with short-acting β2-agonists [AHR may also be related to a lower release of relaxant agonists either neurotransmitters or inflammatory mediators . Physiolagonists . A loweragonists . The secagonists . Howeveragonists . By contagonists . Other cper se. In fact, IL-5 induces in vitro AHR to acetylcholine in isolated rabbit ASM tissue and IL-13 reduces responsiveness to adrenaline in human ASM cells [in vitro using human bronchi from actively sensitized [via its enzymatic activity. However, such AHR occurs after an initial calcium response and, thus, in the absence of any spontaneous contraction. Indeed, AHR may also depend on excitation/contraction coupling, which associates two subsequent steps within the ASM cell, that is, (i) the calcium response induced by extracellular messengers and (ii) the calcium sensitivity of the contractile apparatus. Cytokines, such as IL-5 or IL-13, that are both increased in asthma, do not contract ASM SM cells , 58. Mornsitized  or nonsensitized  possiblyα (TNFSF2) [β, can increase CD38 (ADP-ribosyl cyclase) expression. Then, CD38 increases cyclic ADP-ribose (cADPR) production, which, in turns, activates ryanodin receptor (RyR), leading to an increase in cytosolic calcium concentration [On the one hand, following extracellular stimulation, ASM cytosolic calcium homeostasis is dependant on various components. For instance, the spontaneous return to baseline of the cytosolic calcium concentration is dramatically delayed in asthmatic ASM cells . Such a (TNFSF2) . Alternantration –66. In antration , leadingα (C/EBP-α) expression is decreased within the asthmatic ASM [α binding sites, such a deficit in C/EBP-α has been proposed to account for the increased expression of MLCK [ε inhibits calponin, an actin thin filament-associated protein [α (TNFSF2), which activates RhoA [On the other hand, the calcium sensitivity of the contractile apparatus can be increased, leading to a higher contractile response after a similar ASM intracellular calcium rise . Two difatic ASM . Since t of MLCK . Neverth of MLCK , 74. Reg protein  that dec protein . It also protein  that als protein . Alterna protein , and alstes RhoA , or IL-1tes RhoA .in vitro is directly related to ASM mass in human bronchial ring preparations [Finally, the alteration in the dynamic properties of ASM in asthma is still a matter of current debate. The maximal velocity of shortening in ASM from sensitized animals is significantly greater than those in nonsensitized  and may arations , force garations . Like any muscle, ASM may overcome loads that normally moderate ASM shortening . Two maiIn asthma, several mechanisms may account for an alteration in the contraction/load coupling , 98 suchOn the other hand, thickening of the airway wall and the enhanced amount of ECM also increase airway stiffness and decrease airway compliance  that migWhile early studies regarding ASM function in asthma focussed on its contractile properties, a growing body of evidence now consistently demonstrates that ASM is no longer a sole target of the inflammatory process. Indeed, ASM cell also displays proinflammatory and immunomodulatory functions , 116, thThere is evidence that the asthmatic ASM layer is infiltrated by an increased number of mast cells , 140–147α (TNFSF2), which stimulate the production of TGF-β1 and, to a lesser extent, SCF by ASM cells, which in turns, induce mast cell chemotaxis [3CL1 [3CL1, the additional presence of VIP is necessary [The mechanism of such a myositis has been firstly related to the production of mast cell chemotactic factors by the ASM itself, through an autoactivation loop  Figure . Indeed,emotaxis . Moreoveemotaxis , Th2 [13emotaxis , 155 or emotaxis . For insemotaxis , CXCL8 [in vitro findings are in agreement with ultrastructural analysis of asthmatic ASM using electron microscopy, demonstrating the absence of direct cell-cell contact between ASM and mast cells [ Once present within the ASM bundle, mast cells can adhere to ASM. This adhesion has been initially ascribed to a cell-cell interaction involving an Ig superfamily member, that is, cell adhesion molecule 1 (CADM1), previously known as tumor suppressor in lung cancer 1 (TLSC-1 ) , 157 Fi. Howeverst cells .TC phenotype, containing both tryptase and chymase [The majority of mast cells infiltrating the asthmatic ASM layer are typically of the MC chymase , 159. In chymase , 143. Su chymase . However chymase . Mast ce chymase . However chymase , 162, fo chymase . Taking into account the following characteristics: (i) microlocalization of mast cells within the ASM layer, (ii) mast cell adherence to the ASM, and (iii) mast cell activation within the ASM, it is tempting to consider that a close functional relationship may exist between these two cell types. in vitro [in vivo [β1 secretion, which, in turns, promotes ASM cells differentiation towards a contractile phenotype, characterized by an increase expression of α-actin and enhanced contractility [α-smooth muscle actin [On the one hand, mast cells alter functional and phenotypic properties of ASM cells. Indeed, mast cell-derived mediators contribute to AHR and ASM remodelling . For insin vitro  or in vi[in vivo . Tryptasactility . Interesactility , 145, anle actin . Mast cell myositis may also promote ASM remodelling. However, although several mast cell products such as tryptase are known to individually stimulate both DNA synthesis and ASM proliferation , cocultuOn the other hand, ASM cells alter functional and phenotypic properties of mast cells. Indeed, ASM cells can promote mast cell survival, hence providing a favorable microenvironnement for mast cells . They al+ T cell microlocalization within ASM layer was first reported in an elegant experimental rat asthma model [in vitro [ex vivo [Up to now, only few studies have documented T cell infiltration within the ASM layer in asthma. CD4ma model . This fima model , 15 and ma model , 140. Homa model . In thisin vitro . Nonethe[ex vivo , 15, henin vitro to resting ASM cells from nonasthmatic patients and that such an adhesion was enhanced when ASM cells were primed with proinflammatory cytokines such as TNF-α (TNFSF2) [via their MHC class II [ex vivo suggesting VCAM-1 mediated intercellular signalling, the role of the above molecules in adherence between T cell and ASM cell from asthmatic patients has not yet been considered. Two other ASM cell-surface molecules, CD40 (TNFRSF5) [Lazaar and coworkers originally demonstrated that activated T cells can adhere (TNFSF2) . These f(TNFSF2) . This ad(TNFSF2) . More reclass II . Althougclass II , 120, thTNFRSF5) , 169–171TNFRSF5) , both exTNFRSF5) , 174. Ho+ T cells from OVA-sensitized rats increases ASM mass, which is both associated with an increased ASM proliferation and decreased apoptosis ex vivo [in vitro only upon direct CD4+ T cells-ASM cells contact, highlighting the need for close cellular interaction between these two cell types [in vitro a role for T cell in ASM DNA synthesis and proliferation in ASM cells from nonasthmatic patients [Close interaction between T cells and ASM cells may lead to stimulatory cross-talk between these two cell types, but little is known about the functional consequences of such an interaction. On the one hand, T cells may alter functional properties of ASM cells. In this connection, T cells alter ASM contractile phenotype, enhancing ASM contractility to acetylcholine and reducing its relaxation to isoproterenol in isolated rabbit ASM tissue . T cells ex vivo . Such anll types . These opatients . The rolpatients . Moreovepatients . Collect+ T cells and ASM cells also enhances T cell survival, thus possibly contributing to the perpetuation of bronchial inflammation [via their MHC class II molecules to resting CD4+ T cells [+ T cells activation, adherence between these cells, and finally release of IL-13, that, in turns, leads to increase in the contractile response to acetylcholine of isolated rabbit ASM tissues [On the other hand, ASM cells may also alter functional properties of T cells. Indeed, a direct contact between CD4ammation . Moreove T cells , which l tissues . β [ASM remodelling represents a key feature of asthmatic bronchial remodelling  Figure . Indeed,β , which iβ . The mechanisms underlying ASM remodelling involve ASM hyperplasia, ASM hypertrophy, and an excessive deposition of proteins of the ECM .Three mechanisms may account for ASM hyperplasia, that is, (i) ASM cell proliferation, (ii) reduced ASM cell apoptosis, and more recently (iii) migration of myofibroblasts within the ASM layer.in vitro [in vivo [in vivo was a matter of debate since the number of PCNA or Ki67 positive ASM cells was unchanged in asthmatic bronchial samples [Compelling evidence now suggests an excessive ASM cell proliferation in asthma both in vitro , 180 and[in vivo , 181. Ho[in vivo , 181, th samples . The reain vitro, including growth factors, cytokines, chemokines, inflammatory mediators, enzymes, ECM components, reactive oxygen species, and mechanical stress (G proteins (GPCR) [A wide array of mitogenic factors are known to promote ASM cell proliferation l stress . Such mis (GPCR) . Upon acs (GPCR) .in vitro than nonasthmatic ASM cells irrespective of extracellular stimuli [α were found in asthmatics and could explain enhanced proliferation [α expression is impaired [γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF-1), and mitochondrial transcription factor A (mtTFA) [via a different mechanism [However, asthmatic ASM cells can also proliferate faster  stimuli , hence sferation , throughimpaired . An alte (mtTFA) . This tr (mtTFA)   . More reechanism . Such meechanism  , Fas (TNFRSF6), TRAILR1 (TNFRSF10A), TRAILR2 (TNFRSF10B) [α (TNFSF2) [in vitro [in vitro. Indeed, activation of peroxisome proliferator-activated receptor γ (PPARγ), which is expressed by ASM cells, by its synthetic ligand commonly used in diabetes treatment, may also induce apoptosis [Cardiotrophin-1 , endotheFRSF10B) , and sti(TNFSF2) , 189, so(TNFSF2) , Fas ant(TNFSF2)  induces (TNFSF2) . ECM-deg(TNFSF2) , which r(TNFSF2) . Conversin vitro . Some trpoptosis . Moreovepoptosis . Howeverex vivo in asthmatic ASM following allergen challenge, suggesting that this cytokine may play a role in asthmatic ASM apoptosis [ex vivo in ASM, in severe asthma, and in case of fatal asthma, respectively [ex vivo, irrespective of asthma severity [In vitro, two reports have also drawn negative results, showing no change in spontaneous apoptosis within asthmatic ASM cells [In asthma, the hypothesis of an imbalance between ASM proliferation and apoptosis is mainly supported by a rat model of T-cell driven remodelling . Furtherpoptosis . Besidesectively . Conversectively  and Benaseverity . In vitrSM cells , 161. AsMore recently, it has been suggested that migration of ASM precursor cells, either located within the bronchial wall or derived from peripheral blood fibroblast progenitors, towards the ASM bundles may also participate in ASM hyperplasia . CellulaMigration of myofibroblasts may participate in ASM hyperplasia, by differentiating to ASM-like cells . Such an in vitro human nonasthmatic ASM cell migration, such as cytokines, growth factors, chemokines, and some ECM components [in vivo. However, the origin of myofibroblasts remains a source of debate and several hypotheses have been put forward. First, myofibroblasts may come from resident fibroblasts that have differentiated  or from mponents   a main effector of AHR, (ii) a proinflammatory and immunomodulatory cell through its synthetic properties and its expression of a wide range of cell surface molecules, and (iii) a leading cell involving in bronchial remodelling. If AHR and bronchial inflammation respond fairly well to conventional therapy such as bronchodilators and anti-inflammatory drugs, ASM remodelling remains insensitive to these treatments . Targeti