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Background: Recombinant human calcineurin B subunit (rhCNB) has been shown to be an immune‐stimulatory protein promoting cytokine production and inducing phenotypic maturation of Dendritic cells (DCs). In vivo, it has good antitumor efficacy, and has potential as an antitumor drug. Exogenous rhCNB was found to be internalized into tumor cells via the Toll‐like receptor 4 (TLR4) complex, but it was not known whether its immuno‐modulatory and antitumor functions involved entry by this same route. Methods: The production and secretion of the cytokines and chemokines in innate immune cells induced by rhCNB were determined by ELISA, and the expression of CD40, CD80, CD86, and MHCII was analyzed by FACs. Experimental Lewis lung cancer (LLC) model was prepared in C57 BL/6 wild‐type (WT) mice, TLR4−/− mice or their littermates by the inoculation of LLCs in their right armpit, and then administrated daily intraperitoneal injections (0.2 mL) of normal saline, rhCNB 20 mg/kg, and rhCNB 40 mg/kg, respectively. Results: Recombinant human calcineurin B subunit promoted the production of antitumor cytokines by innate immune cells, and culture supernatants of rhCNB‐stimulated immune cells induced apoptosis of LLCs. In addition, rhCNB up‐regulated CD40, CD80, CD86, and MHCII expression in macrophages and DCs in TLR4+ cells but failed to do so in TLR4 deficient cells. rhCNB also induced the formation of CD4+ and CD8+T cells in splenocytes from WT mice, but not from TLR4‐deficient littermates. Intraperitoneal administration of WT C57BL/6 mice with rhCNB resulted in a 50% reduction in LLC tumor growth, but failed to inhibit tumor growth in TLR4−/− littermates. Conclusions: The in vivo antitumor and immunomodulatory effects of rhCNB are mediated by the TLR4. This conclusion is important for the further understanding and development of rhCNB as an antitumor drug. [ABSTRACT FROM AUTHOR]

ABSTRACT: Aim: Arteriolar hyalinosis (AH) is a common lesion in allograft biopsies taken following kidney transplantation. Recent studies have shown that severe AH may predict transplant outcomes and provide information about previous exposure to certain drugs, such as calcineurin inhibitors (CNI). However, the incidence of AH as a direct result of diabetic nephropathy (DN) after kidney transplantation has not been fully evaluated. This study aimed to assess the impact of primary DN on the development of AH lesions in patients who underwent kidney transplantation. Methods: Eighty‐three patients who underwent living‐donor kidney transplantation between April 2005 and June 2015 were enrolled in this study. A total of 33 patients had DN prior to transplantation. Allograft biopsies were scored according to the Banff classification, and the relationship between the individual histological lesions and clinical baseline data was assessed. Results: At early biopsy (3–12 months), there were no differences in the rates of AH lesions between the DN group and the non‐DN group (ah ≥ 1: 37% vs. 41.3%, P = 0.719; aah ≥ 1: 14.8% vs. 6.5%; P = 0.453). However, there were significant differences between the groups in biopsies taken more than 3 years after the transplant (ah ≥ 2: 83.3% vs. 36.8%, P = 0.013; aah ≥ 2: 66.7% vs. 21.1%, P = 0.011). Multivariable analysis showed that both the length of time after transplantation and the presence of DN were independent risk factors for ah ≥ 2 (odds ratio [OR]: 2.55, 95% confidence interval [CI]: 1.47–19.54, P = 0.011) and aah ≥ 2 (OR: 7.55, 95% CI: 1.49–38.33, P = 0.015). Conclusion: This is the first report showing that the presence of primary DN disease contributes to the development of severe AH late in the course after kidney allografts. [ABSTRACT FROM AUTHOR]

The expression of transient receptor potential canonical 6 (TRPC6) in central nervous system (CNS) is important for neuronal functions and certain neural disorders. However, the regulatory mechanism of TRPC6 expression in neurons is still obscure. In this study, we show that TRPC6 expression in the primary cultured cortical neurons is bidirectionally regulated by glutamate. Activation of NR2A-containing NMDARs induces TRPC6 transcription through a calcineurin-dependent pathway. In contrast, activation of NR2B-containing NMDARs causes TRPC6 degradation through calpain. Thus, TRPC6 expression in neurons is regulated by glutamate in a bidirectional manner that is dependent on NR2A and NR2B. [ABSTRACT FROM AUTHOR]

Introduction Tuberculosis ( TB) is an important cause of morbidity and mortality in renal transplant recipients. Immunosuppressive drugs are one of the most important risk factor for post-transplant tuberculosis ( PTTB). A paucity of data exists about the impact of the type of calcineurin inhibitor on PTTB. Methods In this retrospective study, all adult patients on calcineurin inhibitor-based immunosuppression were included. Patients receiving TB chemoprophylaxis were excluded. Diabetes, duration of dialysis, hepatitis B and C, past treated TB, induction therapy, type of antimetabolite, acute rejection, new onset of diabetes after renal transplantation ( RT) ( NODAT) and cytomegalovirus ( CMV) were analyzed in tacrolimus (Tac) and cyclosporine (CsA) groups. Primary outcome was incidence of TB and secondary outcomes were timeline of development of TB after RT and pattern of TB in the two groups. Results Of the 1664 patients included, 582 patients received CsA-based immunosuppression while 1082 received Tac-based immunosuppression. Duration of dialysis, positive tuberculin skin test, use of induction, mycophenolate mofetil use, CMV infection, and NODAT were significantly more, and hepatitis B infection, past treated TB, and acute rejection episodes were significantly less in the Tac group. At the end of follow-up, incidence of TB in the Tac group was significantly less than in the CsA group (6.1% vs 19.9%, P<.001). Mean time for development of TB after RT was similar in both the groups and nodal and disseminated TB were more common in the Tac group. Conclusion In conclusion, our study shows that use of Tac as compared to CsA significantly decreases incidence of PTTB. Time of infection since transplant was similar in both the groups. However, nodal and disseminated TB were more common in the Tac group. [ABSTRACT FROM AUTHOR]

Fluorescence complementation ( FC) techniques are expedient for analyzing bimolecular protein-protein interactions. Here we aimed to develop a method for visualization of ternary protein complexes using dual-color trimolecular fluorescence complementation (Tri FC)., Dual-color TriFC combines protein fragments of mCherry and mVenus, in which a scaffold protein is bilaterally fused to C-terminal fragments of both fluorescent proteins and combined with potential interacting proteins fused to an N-terminal fluorescent protein fragment. For efficient visual verification of ternary complex formation, TriFC was combined with a cytoplasm to plasma membrane translocation assay., Modular vector sets were designed which are fully compatible with previously reported bimolecular fluorescence complementation (Bi FC) vectors. As a proof-of-principle, the ternary complex formation of the PP2B protein phosphatase Calcineurin-A/Calcineurin-B with Calmodulin-2 was investigated in transiently transformed Nicotiana benthamiana leaf epidermal cells. The results indicate a Calcineurin-B-induced interaction of Calmodulin-2 with Calcineurin-A., Tri FC and the translocation of Tri FC complexes provide a novel tool to investigate ternary complex formations with the simplicity of a Bi FC approach. The robustness of FC applications and the opportunity to quantify fluorescence complementation render this assay suitable for a broad range of interaction analyses. [ABSTRACT FROM AUTHOR]

This viewpoint presents a unifying concept for the treatment of atopic dermatitis ( AD) that is based on the improvement of deficient Notch signalling, which appears to represent the fundamental epithelial defect of AD resulting in epidermal and immunological barrier dysfunction. One study of AD patients demonstrated a marked epidermal deficiency of Notch receptors and several mouse models with genetically suppressed Notch signalling exhibit dry skin, signs of scratching, skin barrier abnormalities, increased transepidermal water loss and Th2 cell-mediated immunological changes closely resembling human AD. Notch signalling is critically involved in the differentiation of regulatory T cells, in the feedback inhibition of activated innate immunity, in the repression of activating protein-1 ( AP-1), the regulation of late epidermal differentiation associated with filaggrin- and stratum corneum barrier lipid processing, in aquaporin 3- and claudin-1 expression and in keratinocyte-mediated release of thymic stromal lymphopoietin ( TSLP), which promotes Th2-driven immune responses with TSLP- and IL-31-mediated stimulation of cutaneous sensory neurons involved in the induction of itch. Translational evidence will be provided that all major therapeutic regimens employed for the treatment of AD such as glucocorticoids, calcineurin inhibitors and UV radiation may converge in the upregulation of impaired Notch signalling, the proposed pathogenic defect of AD. [ABSTRACT FROM AUTHOR]

Aims Calcineurin inhibitors ( CNIs) taken after heart transplantation lead to excellent short-term outcomes, but long-term use may cause chronic nephrotoxicity. Our aim was to identify, appraise, select and analyse all high-quality research evidence relevant to the question of the clinical impact of CNI-sparing strategies in heart transplant patients. Methods We carried out a systematic review and meta-analysis of randomized controlled trials on CNI reduction in heart transplant recipients. Primary outcomes were kidney function and acute rejection after 1 year. Secondary outcomes included graft loss, all-cause mortality and adverse events. Results Eight open-label studies were included, with 723 patients (four tested de novo CNI reduction and four maintenance CNI reduction). Calcineurin inhibitor reduction did not improve creatinine clearance at 12 months 5.46 [−1.17, 12.03] P = 0.32 I2 = 65.4%. Acute rejection at 12 months (55/360 vs. 52/332), mortality (18/301 vs. 15/270) and adverse event rates (55/294 vs. 52/281) did not differ between the low- CNI and standard- CNI groups. There was significant benefit on creatinine clearance in patients with impaired renal function at 6 months [+12.23 (+5.26, +18.82) ml min−1, P = 0.0003] and at 12 months 4.63 [−4.55, 13.82] P = 0.32 I2 = 75%. Conclusions This meta-analysis did not demonstrate a favourable effect of CNI reduction on kidney function, but there was no increase in acute rejection. To provide a better analysis of the influence of CNI reduction patterns and associated treatments, a meta-analysis of individual patient data should be performed. [ABSTRACT FROM AUTHOR]

Transmembrane α-amino-3-hydroxy-5-methyl-4-isoxazole propionate ( AMPA) receptor regulatory proteins ( TARPs) are auxiliary subunits that regulate AMPA receptor trafficking to the plasma membrane and localization to postsynaptic sites. The classical TARP family consists of four members: stargazin/γ-2, γ-3, γ-4 and γ-8. The TARP γ-8 isoform, which is highly expressed in the hippocampus, has a unique, long C-terminal domain with five distinct regions: two glycine-rich regions, a serine/arginine-rich region, a proline/alanine ( P/ A) rich region, and a PSD-95/ Dlg/ ZO-1 ( PDZ) binding motif. We performed mass spectrometry and immunoprecipitation assays to identify specific binding partners for the γ-8 C-terminal tail and found that γ-8, but not stargazin/γ-2, co-immunoprecipitated with calcineurin/ PP2 B, a Ca2+/calmodulin-dependent Ser/ Thr phosphatase. Co-immunoprecipitation and immunoblot analyses of lysates from COS-7 cells co-transfected with calcineurin and either wild type or chimeric γ-8 revealed that a section of the C-terminal tail (residues 356-421) can bind calcineurin. Futhermore, γ-8 lacking the P/ A-rich region (residues 383-399) did not bind to calcineurin. In addition, the GST-γ-8 C-terminal tail (residues 353-414) fusion protein containing the P/ A-rich region bound to purified calcineurin in a Ca2+/calmodulin-dependent manner, whereas GST-γ-8 with a deletion of the P/ A-rich region did not. Peptide competition assays demonstrated that γ-8 may interact with the hydrophobic pocket defined by β-sheet 14 and/or adjacent regions of the catalytic A subunit of calcineurin. These results indicate that the γ-8 P/ A-rich region is essential for binding calcineurin, suggesting that the γ-8/calcineurin complex may regulate AMPA receptor phosphorylation and trafficking. Structured digital abstract with by () to by () with by () with and by () with , and by () with and by () with and by (, , ) with by () with , , and by () with by (), and by () [ABSTRACT FROM AUTHOR]

Calcineurin B‐like protein‐interacting protein kinases (CIPKs) are components of Ca2+ signaling in responses to abiotic stresses. In this work, the full‐length cDNA of a novel CIPK gene (TaCIPK14) was isolated from wheat and was found to have significant sequence similarity to OsCIPK14/15. Subcellular localization assay revealed the presence of TaCIPK14 throughout the cell. qRT‐PCR analysis showed that TaCIPK14 was upregulated under cold conditions or when treated with salt, PEG or exogenous stresses related signaling molecules including ABA, ethylene and H2O2. Transgenic tobaccos overexpressing TaCIPK14 exhibited higher contents of chlorophyll and sugar, higher catalase activity, while decreased amounts of H2O2 and malondialdehyde, and lesser ion leakage under cold and salt stresses. In addition, overexpression also increased seed germination rate, root elongation and decreased Na+ content in the transgenic lines under salt stress. Higher expression of stress‐related genes was observed in lines overexpressing TaCIPK14 compared to controls under stress conditions. In summary, these results suggested that TaCIPK14 is an abiotic stress‐responsive gene in plants. [ABSTRACT FROM AUTHOR]

Highlights: [•] Green light provided protection against the deleterious effects of UV irradiation upon yeast cells. [•] High density green photons protected the yeast cells against UV in synergism with unfolded protein response inducers. [•] Photoprotective action of green light against UV required intact UPR pathway. [•] The cnb1Δ cells, lacking the regulatory subunit of calcineurin, became more sensitive to UV after green light exposure. [ABSTRACT FROM AUTHOR]

Background Most psoriasis patients suffering from mild to moderate disease are treated with first-line topical treatments, including corticosteroids, vitamin D analogues, topical retinoids and calcineurin inhibitors. Although evidence-based guidelines on combinations are lacking, the majority of patients will be treated with combinations of these popular topicals at some point during their life-long treatment. Objectives We intended to systematically review all available literature on the efficacy and safety of combinations of first-line topicals in chronic plaque psoriasis, and ultimately, to propose recommendations for combined regimens concerning first-line treatments. Methods Databases used as data sources were PubMed, EMBASE and the Cochrane Controlled Clinical Trial Register. According to standardized procedures, literature searches were performed and a level of evidence was determined. Results In total, 2918 publications on topical combination therapy were revealed, of which 45 articles on first-line combinations were included. The combination of potent and superpotent corticosteroids with vitamin D analogues provides an improvement of psoriasis within 2 weeks, reaching a maximal improvement after 4 weeks in the majority of patients. The two-compound product permits once-daily treatment and therefore is a good solution for chronic plaque psoriasis including scalp psoriasis. Combinations of corticosteroids, with tazarotene or with calcineurin inhibitors do not provide an advantage above corticosteroid monotherapy. Conclusion The combination of potent corticosteroids with calcipotriol has been studied most extensively and should be regarded as an efficacious and safe treatment option with the two-compound products as the most practical solution. [ABSTRACT FROM AUTHOR]

Summary Calcineurin (CN) is an important phosphatase that mediates many physiological and pathological processes. The regulators of calcineurin (RCAN1) and Cu, Zn superoxide dismutase (SOD1) are two endogenous modulators of CN activity. Cyclosporine A (CsA) is a well-known exogenous inhibitor of CN and used as an immunosuppressive drug after transplantation and for the treatment of immune diseases. The degree of CN inhibition by CsA varies among each tissue. The brain accumulates low levels of CsA due to the blood-brain barrier after oral administration. In our study, we investigated RCAN1 and SOD1 expression in long-term CsA-treated mouse brain. Using Western blot, we found that chronic CsA treatment had caused significant up-regulation of RCAN1-1L and RCAN1-4 protein isoforms after 25 days in mouse brain. At the same time, chronic CsA treatment also resulted in decreased expression of SOD1. We simultaneously found more dramatic CN inhibition in mouse brain. It was suspected that the significant reduction of CN activity in vivo resulted partially from up-regulated RCAN1 and down-regulated SOD1 expression. In contrast, CsA treatment in SY5Y cells affected SOD1 expression and CN activity significantly, but had no obvious effects on RCAN1-1 mRNA expression. The changes of RCAN1, SOD1, and CN activity may be part of maladaptive responses, resulting in neuropathological conditions. These data might partially explain CsA neurotoxicity despite the low concentration of CsA in brain. © 2013 IUBMB Life 65(5):445-453, 2013. [ABSTRACT FROM AUTHOR]

HUS is a well-known entity primarily associated with bacterial infection and is characterized by a classic triad of anemia, thrombocytopenia, and kidney injury. Its atypical form has been associated with calcineurin inhibitors and has been extensively discussed in renal transplantation. We present a case of tacrolimus-associated HUS in a pediatric heart transplant recipient, which we believe to be previously unreported in the literature. [ABSTRACT FROM AUTHOR]

The article discusses research on the effectiveness of tacrolimus 0.1% ointment in the treatment of allergic contact dermatitis (ACD). The macrolactam immunosuppressant has been shown to prevent the calcineurin expression of genes for producing cytokine production and activating T lymphocyte. Findings demonstrated tacrolimus as safe and effective for ACD treatment.