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Background: This systematic review aims to evaluate and synthesize evidence on the efficacy and safety of topical treatments for VKC, focusing on the effectiveness of treatments like cyclosporine A, tacrolimus, and other topical therapies in alleviating symptoms and improving clinical outcomes. Methods: The review followed PRISMA 2020 guidelines, including studies published in English from 2014 to 2024. Results: Eight studies met the inclusion criteria. The review found that both cyclosporine A and tacrolimus significantly reduced symptom and sign scores in VKC patients. Rebamipide was comparable in efficacy to cyclosporine and tacrolimus. Tacrolimus showed efficacy in corticosteroid-refractory cases, while cyclosporine A reduced steroid dependency and related side effects. Conclusion: Topical cyclosporine A and tacrolimus are effective in managing VKC, with improvements in symptoms and clinical signs observed across studies. Cyclosporine A is particularly effective in reducing reliance on corticosteroids, while tacrolimus is beneficial for corticosteroid-refractory VKC. Personalized treatment strategies should be considered based on individual patient responses and tolerability.

Background: This systematic review aims to evaluate and synthesize evidence on the efficacy and safety of topical treatments for VKC, focusing on the effectiveness of treatments like cyclosporine A, tacrolimus, and other topical therapies in alleviating symptoms and improving clinical outcomes. Methods: The review followed PRISMA 2020 guidelines, including studies published in English from 2014 to 2024. Results: Eight studies met the inclusion criteria. The review found that both cyclosporine A and tacrolimus significantly reduced symptom and sign scores in VKC patients. Rebamipide was comparable in efficacy to cyclosporine and tacrolimus. Tacrolimus showed efficacy in corticosteroid-refractory cases, while cyclosporine A reduced steroid dependency and related side effects. Conclusion: Topical cyclosporine A and tacrolimus are effective in managing VKC, with improvements in symptoms and clinical signs observed across studies. Cyclosporine A is particularly effective in reducing reliance on corticosteroids, while tacrolimus is beneficial for corticosteroid-refractory VKC. Personalized treatment strategies should be considered based on individual patient responses and tolerability.

Drug loading into mesoporous carriers may help to improve the dissolution of poorly aqueous-soluble drugs. However, both preparation method and carrier properties influence loading efficiency and drug release. Accordingly, this study aimed to compare two preparation methods: formulation into liquisolid systems (LSS) and co-milling for their efficiency in loading the poorly soluble model drug cyclosporine A (CyA) into mesoporous magnesium aluminometasilicate Neusilin® US2 (NEU) or functionalized calcium carbonate (FCC). Scanning electron microscopy was used to visualize the morphology of the samples and evaluate the changes that occurred during the drug loading process. The solid-state characteristics and physical stability of the formulations, prepared at different drug concentrations, were determined using X-ray powder diffraction. In vitro release of the drug was evaluated in biorelevant media simulating intestinal fluid. The obtained results revealed improved drug release profiles of the formulations when compared to the milled (amorphous) CyA alone. The dissolution of CyA from LSS was faster in comparison to the co-milled formulations. Higher drug release was achieved from NEU than FCC formulations presumably due to the higher pore volume and larger surface area of NEU.

Cyclosporine A (CsA) is used for the treatment of dry eye (DE) with good clinical results, improving tear secretion and decreasing subjective symptoms. These effects are attributed to the improved tear film dynamics, but there are no data on the effect of CsA on the abnormal sensory nerve activity characteristic in DE. Our purpose was to evaluate the CsA effect on the enhanced activity of corneal cold thermoreceptors in a tear-deficient DE animal model using in vitro extracellular recording of cold thermoreceptors nerve terminal impulses (NTIs) before and in the presence of CsA. NTI shape was also analyzed. Blinking frequency and tearing rate were also measured in awake animals before and after topical CsA. CsA increased the tearing and blinking of treated animals. CsA significantly decreased the peak response to cold of cold thermoreceptors. Neither their spontaneous NTIs discharge rate nor their cooling threshold were modified. CsA also seemed to reverse some of the changes in NTI shape induced by tear deficiency. These data suggest that, at least in part, the beneficial clinical effects of CsA in DE can be attributed to a direct effect on sensory nerve endings, although the precise mechanisms underlying this effect need further studies to be fully clarified.

Pre-clinical studies have indicated that mitoprotective drugs may add cardioprotection beyond rapid revascularization, antiplatelet therapy and risk modification. We review the clinical efficacy of mitoprotective drugs that have progressed to clinical testing comprising cyclosporine A, KAI-9803, MTP131 and TRO 40303. Whereas cyclosporine may reduce infarct size in patients undergoing primary angioplasty as evaluated by release of myocardial ischaemic biomarkers and infarct size imaging, the other drugs were not capable of demonstrating this effect in the clinical setting. The absent effect leaves the role of the mitochondrial permeability transition pore for reperfusion injury in humans unanswered and indicates that targeting one single mechanism to provide mitoprotection may not be efficient. Moreover, the lack of effect may relate to favourable outcome with current optimal therapy, but conditions such as age, sex, diabetes, dyslipidaemia and concurrent medications may also alter mitochondrial function. However, as long as the molecular structure of the pore remains unknown and specific inhibitors of its opening are lacking, the mitochondrial permeability transition pore remains a target for alleviation of reperfusion injury. Nevertheless, taking conditions such as ageing, sex, comorbidities and co-medication into account may be of paramount importance during the design of pre-clinical and clinical studies testing mitoprotective drugs.

Background: Gingival hyperplasia could occur after the administration of cyclosporine A. Up to 90% of the patients submitted to immunosuppressant drugs have been reported to suffer from this side effect. The role of fibroblasts in gingival hyperplasia has been widely discussed by literature, showing contrasting results. In order to demonstrate the effect of cyclosporine A on the extracellular matrix component of fibroblasts, we investigated the gene expression profile of human fibroblasts after cyclosporine A administration. Materials and methods: Primary gingival fibroblasts were stimulated with 1000 ng/mL cyclosporine A solution for 16 h. Gene expression levels of 57 genes belonging to the “Extracellular Matrix and Adhesion Molecules” pathway were analyzed using real-time PCR in treated cells, compared to untreated cells used as control. Results: Expression levels of different genes were significantly de-regulated. The gene CDH1, which codes for the cell adhesion protein E-cadherin, showed up-regulation. Almost all the extracellular matrix metalloproteases showed down-regulation (MMP8, MMP11, MMP15, MMP16, MMP24, MMP26). The administration of cyclosporine A was followed by down-regulation of other genes: COL7A1, the transmembrane receptors ITGB2 and ITGB4, and the basement membrane constituents LAMA2 and LAMB1. Conclusion: Data collected demonstrate that cyclosporine inhibits the secretion of matrix proteases, contributing to the accumulation of extracellular matrix components in the gingival connective tissue, causing gingival overgrowth. Patients affected by gingival overgrowth caused by cyclosporine A need to be further investigated in order to determine the role of this drug on fibroblasts.

Transplantacija bubrega je najpoželjniji metod lečenja pedijatrijskih pacijenata sa terminalnom bubrežnom insuficijencijom, a Ciklosporin A (CsA) je imunosupresivni lek koji se koristi kod transplantiranih bolesnika u cilju prevencije odbacivanja kalema. Različita ekspresija komponenata familije enzima CYP3A (prvenstveno CYP3A4 i CYP3A5), POR enzima i P-glikoproteina može biti uzrok individualne varijabilnosti u apsorpciji, metabolizmu i tkivnoj distribuciji CsA. Individualni odgovor na terapiju može biti uslovljen prisustvom različitih alelnih varijanti (polimorfizama tipa zamene jednog nukleotidnog ponovka , eng. single nucleotide polymorphisms-SNP) u genima koji kodiraju navedene enzime i transporter. Takođe, biološko sazrevanje dečijeg organizma doprinosi razlikama u farmakokinetici CsA u grupi pedijatrijskih pacijenata. Ciljevi studije: 1. da se utvrdi frekvencija alela i genotipova polimorfizama u genima za ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) i POR (rs1057868) kod pedijatriskih pacijenata sa transplantiranim bubregom u Srbiji, 2. da se ispita uticaj uzrasta pacijenta, kao nezavisnog faktora, na farmakokinetiku CsA tokom prve godine nakon transplantacije, 3. da se ispita uticaj polimorfizama u genima za ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) i POR (rs1057868) na farmakokinetiku CsA kod pedijatrijskih pacijenata sa transplantiranim bubregom u Srbiji, tokom prve godine nakon transplantacije, 4. da se ispita uticaj polimorfizama u genima za ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) i POR (rs1057868) u zavisnosti od uzrasta pacijenta, na farmakokinetiku CsA kod pedijatrijskih pacijenata sa transplantiranim bubregom u Srbiji, tokom prve godine nakon transplantacije. Metodologija: Istraživanje je obavljeno na Univerzitetskoj dečjoj klinici, u Centru za hemodijalizu i transplantaciju, jedinom mestu u Srbiji, gde se radi transplantacija bubrega kod pedijatrisjkih pacijenata uz njihovo postransplantaciono praćenje, Kidney transplantation is the most desirable method for the treatment of children with terminal renal insufficiency and Cyclosporine A (CsA) is a cornerstone of immunosuppressive therapy used in transplanted patients, in order to prevent the rejection of kidney. Different expression of components of the CYP3A enzyme family (primarily CYP3A4 and CYP3A5), POR enzyme and of P-glycoprotein, may be the cause of individual variability in absorption, metabolism and tissue distribution of CsA. Individual response to therapy may be conditioned by the presence of various allelic variants (single nucleotide polymorphisms-SNP) in genes encoding said enzymes and a transporter. Also, biological maturation of child organism contributes to differences in the pharmacokinetics of CsA in a group of pediatric patients. Aim: 1.to determine the frequency of alleles and genotypes of polymorphisms in genes for ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) and POR (rs1057868) in children with kidney transplants in Serbia; 2. to investigate the effect of polymorphisms in the genes of ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) and POR (rs1057868) on the pharmacokinetics of CsA in children with kidney transplants in Serbia; 3. to consider the influence of child age as an independent factor on the pharmacokinetics of CsA in children with kidney transplants in Serbia; 4. to investigate the effect of polymorphisms in the genes of ABCB1 (rs1045642), CYP3A5 (rs776746), CYP3A4 (rs35599367) and POR (rs1057868) on the age of the patient, on the pharmacokinetics of CsA in children with kidney transplants in Serbia. Methodology: The research was carried out at the University Children's Hospital, at the Center for Hemodialysis and Transplantation, the only place in Serbia, where kidney transplants are performed, in line with post-transplant monitoring. The study included 47 children who had a kidney transplant in the period of 2001-2015. Patients received CsA in the after trans

Purpose: Cyclosporine A (CsA) is the most widely used immunosuppressive agent after a hematopoietic stem cell transplantation (HSCT). Although recommendations for CsA dose conversion from intravenous to oral administration differ from 1:1 to 1:3, most studies did not consider the role of azole antifungals as an important confounder. Therefore, we assess the optimal conversion rate of CsA from intravenous to oral administration in HSCT recipients, taking into account the concomitant use of azole antifungals. Methods: We retrospectively included patients from a large database of 483 patients who underwent a HSCT and received intravenous CsA as part of the conditioning regimen and peritransplant immunosuppression. All patients were converted from intravenous to oral administration in a 1:1 conversion rate. We collected for each patient three CsA trough concentrations during intravenous and oral administration, directly before and after conversion to oral administration. Results: We included 71 patients; 50 patients co-treated with fluconazole, 10 with voriconazole, and 11 without azole co-medication. In patients with voriconazole, the dose-corrected CsA concentration (CsA concentration divided by CsA dosage) was not different between intravenous and oral administration (2.6% difference, p = 0.754), suggesting a CsA oral bioavailability of nearly 100%. In patients with fluconazole and without azole co-medication, the dose-corrected CsA concentration was respectively 21.5% (p < 0.001) and 25.2% (p = 0.069) lower during oral administration. Conclusions: In patients with voriconazole, CsA should be converted 1:1 from intravenous to oral administration. In patients with fluconazole and without azole co-medication, a 1:1.3 substitution is advised to prevent subtherapeutic CsA concentrations.

OBJECTIVE: To study visual pathway pathology detected by visual evoked potentials (VEPs) in patients treated with hematopoietic stem cell transplantation (HSCT) in childhood and to determine the impact of adverse ocular findings, somatic diseases, and conditioning regimens on the VEP results. METHODS: Ophthalmological assessments including pattern VEPs were performed in 47 of 79 patients at a median age of 15 years (range 3-21 years) in median 6 years (1-17 years) after HSCT. Somatic data were extracted from medical records. RESULTS: Eight patients of 47 (17%) demonstrated pathological VEPs with prolonged latencies bilaterally (n = 3) or unilaterally (n = 5) at their latest VEP test at an age of 12-18 years. A subnormal visual acuity was present in 8/11 eyes with pathological VEPs: one eye had cataract, six eyes had cataract surgery where of two had developed secondary cataracts. One eye had residual retinopathy of prematurity. Pathological VEPs were associated with decreased visual acuity (p = 0.00019) but not linked to gender, malignant diagnosis or conditioning. CONCLUSION: VEP recordings showed an association with decreased visual acuity but no relationship with irradiation or chemotherapy in the present study. SIGNIFICANCE: VEP recordings might be of clinical value for children with an unexplained subnormal visual acuity undergoing HSCT.

To advance the development of peptide analogues for improved treatment of pain, we need to learn more about the blood brain barrier transport of these substances. A low penetration into the brain, with an unbound brain to blood ratio, K-p,K-uu, of 0.08, is an important reason for the lack of effect of the enkephalin analogue DAMGO (H-Tyr-D-Ala-Gly-MePhe-Gly-ol) according to earlier findings. The aim of this study was to investigate the role of efflux transporters, metabolism in the brain, and/or elimination through interstitial fluid bulk flow for the brain exposure of DAMGO. The in vivo brain distribution of DAMGO was evaluated using microdialysis in the rat. Data were analyzed with population modeling which resulted in a clearance into the brain of 1.1 and an efflux clearance 14 mu L/min/g_brain. The efflux clearance was thus much higher than the bulk flow known from the literature. Coadministration with the efflux transporter inhibitors cyclosporin A and elacridar in vivo did not affect K-p,K-uu. The permeability of DAMGO in the Caco-2 assay was very low, of the same size as mannitol. The efflux ratio was <2 and not influenced by cyclosporin A or elacridar. These results indicate that the well-known efflux transporters Pgp and Bcrp are not responsible for the higher efflux of DAMGO, which opens up for an important role of other transporters at the BBB.

Background: The transcription factor farnesoid X receptor (FXR) governs bile acid and energy homeostasis, is involved in inflammation, and has protective functions in the liver. In the present study we investigated the effect of Fxr deficiency in mouse precision cut liver slices (PCLS) exposed to a model hepatotoxicant cyclosporin A (CsA). It was anticipated that Fxr deficiency could aggravate toxicity of CsA in PCLS and pinpoint to novel genes/processes regulated by FXR. Methods: To test this hypothesis, PCLS obtained from livers of wild type mice (WT-PCLS) and Fxr-knockout mice (FXRKO-PCLS) were treated with 40 μM CsA for 24 h and 48 h. ATP and histological assays were applied to assess the viability of PCLS. DNA microarrays combined with bioinformatics analysis were used to identify genes and processes that were affected by CsA in WT-PCLS and/or FXRKO-PCLS. In addition, WT-PCLS and FXRKO-PCLS were exposed to the endogenous FXR ligand chenodeoxycholic acid (CDCA) and subjected to q-PCR to determine whether subsets of known FXR-targets and the identified genes were regulated upon FXR activation in an FXR-dependent manner. Results: No difference in viability was observed between WT-PCLS and FXRKO-PCLS upon CsA treatment. Transcriptomics data analysis revealed that CsA significantly upregulated stress-response and inflammation and significantly downregulated processes involved in lipid and glucose metabolism in WT-PCLS and FXRKO-PCLS. However, only in FXRKO-PCLS, CsA upregulated additional pro-inflammatory genes and downregulated genes related to mitochondrial functions. Furthermore, only in WT-PCLS, CDCA upregulated a subset of known FXR-target genes as well as the regulator of inflammation and mitochondrial functions peroxisome proliferator- activated receptor delta (Ppar delta). Conclusions: Although FXR governs energy metabolism, no major differences in response to CsA could be observed between WT-PCLS and FXRKO-PCLS in regulation of processes involved in lipi

The paper is a five year retrospective study of the efficacy of 0.2% Cyclosporine A eye cream in the treatment of chronic autoimmunemediated KCS in dogs. 114 dogs of 32 breeds, both sexes, and different ages with chronic autoimmune-mediated KCS were treated with 0.2% Cyclosporine A in the form of eye cream (Optimmune®, Schering - Plough Animal Health) applied to the lower lid conjunctiva at a dose of 6 mm length of the cream strip every 12 hours, during 90 days. Dogs treated with CsA had an improved STT score, specially dogs which had STT values >5 mm/min at the start. Improvement of the clinical picture and healing of superficial corneal lesions were also recorded., U ovom radu je izneta petogodišnja retrospektivna studija o efikasnosti ciklosporin A kreme za oči u terapiji hroničnog autoimunog keratoconjunctivitis sicca kod pasa. Ukupno 114 pasa, 32 rase, oba pola i različite starosti sa hroničnim autoimunim KCS tretirano je ciklosporinom A u vidu kreme za oči (Optimmune®, Schering-Plough Ainimal Health), aplikovanjem na konjunktivu donjeg kapka u dozi od 6 mm dužine kreme na svakih 12 sati tokom 90 dana. Kod pasa tretiranih sa CsA došlo je do povećanja vrednosti Širmerovog suznog testa, naročito kod pasa koji su na početku imali STT vrednost >5 mm/min. Takođe je došlo i do poboljšanja kliničke slike i izlečenja površinskih kornealnih lezija.

type:Text, Aggressive treatment is necessary for continuous high-range proteinuria in cases of pediatric Henoch-Schönlein purpura nephritis (HSPN) as the long-term prognosis is sometimes poor. Cyclosporine (CyA) has immunosuppressive effects as well as a very selective inhibitory effect on T-helper cell function. Here we report two 7-year-old boys with steroid-resistant HSPN treated with CyA. After diagnosis of HSPN, we treated both patients with methylprednisolone and urokinase pulse therapy (MUT) combined with multiple drugs; however, high-range proteinuria persisted and CyA was added to the treatment regimen. The proteinuria subsequently decreased gradually and pathological findings at the second renal biopsy were improved. Furthermore, neither patient showed any adverse effects, such as hypertension, encephalopathy, or chronic nephrotoxicity, to the CyA treatment.In conclusion, these results suggest that CyA may be a safe and effective treatment for steroid-resistant severe HSPN.

The paper is a five year retrospective study of the efficacy of 0.2% Cyclosporine A eye cream in the treatment of chronic autoimmunemediated KCS in dogs. 114 dogs of 32 breeds, both sexes, and different ages with chronic autoimmune-mediated KCS were treated with 0.2% Cyclosporine A in the form of eye cream (Optimmune®, Schering - Plough Animal Health) applied to the lower lid conjunctiva at a dose of 6 mm length of the cream strip every 12 hours, during 90 days. Dogs treated with CsA had an improved STT score, specially dogs which had STT values >5 mm/min at the start. Improvement of the clinical picture and healing of superficial corneal lesions were also recorded., U ovom radu je izneta petogodišnja retrospektivna studija o efikasnosti ciklosporin A kreme za oči u terapiji hroničnog autoimunog keratoconjunctivitis sicca kod pasa. Ukupno 114 pasa, 32 rase, oba pola i različite starosti sa hroničnim autoimunim KCS tretirano je ciklosporinom A u vidu kreme za oči (Optimmune®, Schering-Plough Ainimal Health), aplikovanjem na konjunktivu donjeg kapka u dozi od 6 mm dužine kreme na svakih 12 sati tokom 90 dana. Kod pasa tretiranih sa CsA došlo je do povećanja vrednosti Širmerovog suznog testa, naročito kod pasa koji su na početku imali STT vrednost >5 mm/min. Takođe je došlo i do poboljšanja kliničke slike i izlečenja površinskih kornealnih lezija.

The glycosaminoglycan (GAG) hyaluronan (HA) is recognized as an important structural component of the extracellular matrix, but it also interacts with cells during embryonic development, wound healing, inflammation, and cancer; i.e., important features in normal and pathological conditions. The specific physico-chemical properties of HA enable a unique hydration capacity, and in the last decade it was revealed that in the interstitium of the renal medulla, where the HA content is very high, it changes rapidly depending on the body hydration status while the HA content of the cortex remains unchanged at very low amounts. The kidney, which regulates fluid balance, uses HA dynamically for the regulation of whole body fluid homeostasis. Renomedullary HA elevation occurs in response to hydration and during dehydration the opposite occurs. The HA-induced alterations in the physicochemical characteristics of the interstitial space affects fluid flux; i.e., reabsorption. Antidiuretic hormone, nitric oxide, angiotensin II, and prostaglandins are classical hormones/compounds involved in renal fluid handling and are important regulators of HA turnover during variations in hydration status. One major producer of HA in the kidney is the renomedullary interstitial cell, which displays receptors and/or synthesis enzymes for the hormones mentioned above. During several kidney disease states, such as ischemia-reperfusion injury, tubulointerstitial inflammation, renal transplant rejection, diabetes, and kidney stone formation, HA is upregulated, which contributes to an abnormal phenotype. In these situations, cytokines and other growth factors are important stimulators. The immunosuppressant agent cyclosporine A is nephrotoxic and induces HA accumulation, which could be involved in graft rejection and edema formation. The use of hyaluronidase to reduce pathologically overexpressed levels of tissue HA is a potential therapeutic tool since diuretics are less efficient in removing water

Tato práce je zaměřena na vývoj nové analytické metody pro stanovení Cyklosporinu A v krvi pacientů po transplantaci a s autoimunitními chorobami., This thesis is focus on the determination of cyclosporine by direct injection mass spectrometry., Katedra biologických a biochemických věd, 1. Prezentace výsledků diplomové práce 2. Diskuze k posudkům vedoucího a oponenta diplomové práce 3. Diplomant zodpověděl všechny dotazy a připomínky k DP

The goal of this project was to understand how cyclophilin A (CypA) regulates the prolactin receptor (PRLr) signaling. The data have demonstrated that CypA, serving as a molecular switch via its peptidyl-prolyl isomerase (PPI) activity, binds to and regulates the function of the PRLr. Furthermore, point mutagenesis analysis has implied that a single prolyl imide bond (Pro-334) within the X-Box motif of intracellular domain of the PRLr is most likely responsible for the observed effects of CypA on the PRLr signaling. Finally, we have conducted several proof-of-concept studies that may lead to novel anti-breast cancer agents., The original document contains color images.

A 47-year-old woman who had been treated for systemic lupus erythematosus (SLE) with myasthenia gravis (MG) was admitted to our hospital with acute onset of severe anemia after administration of isoniazid. Pure red cell aplasia (PRCA) was confirmed by elevated serum iron levels, reticulocytopenia and bone marrow aspiration showing a remarkable reduction of erythroblasts. Finally, cyclosporine A successfully improved PRCA. Although both SLE and MG have the potential complication of PRCA, we report here a case of isoniazid-triggered PRCA., Rheumatology international, 30(12), pp.1643-1645; 2010

Background. Use of the mammalian target of rapamycin inhibitor everolimus with an accompanying reduction in calcineurin inhibitor (CNI) exposure has shown promise in preserving renal function in maintenance thoracic transplant patients, but robust, long-term data are required. Methods. In a prospective, open-label, multicenter study, thoracic transplant recipients more than or equal to 1 year posttransplant with mild-to-moderate renal insufficiency were randomized to continue their current CNI-based immunosuppression or convert to everolimus with predefined CNI exposure reduction. After a 12-month core trial, patients were followed up to month 24 after randomization. Results. Of 245 patients who completed the month 12 visit, 235 patients (108 everolimus and 127 controls) entered the 12-month extension phase. At month 24, mean measured glomerular filtration rate had increased by 3.2 +/- 12.3 mL/min from the point of randomization in everolimus-treated patients and decreased by 2.4 +/- 9.0 mL/min in controls (Pandlt;0.001), a difference that was significant within both the heart and lung transplant subpopulations. During months 12 to 24, 5.6% of everolimus patients and 3.1% of controls experienced biopsy-proven acute rejection (P=0.76). There were no significant differences in the rate of adverse events or serious adverse events (including pneumonia) between groups during months 12 to 24. Conclusions. Converting maintenance thoracic transplant recipients to everolimus with low-exposure CNI results in a renal benefit that is sustained to 2 years postconversion, with significantly improved measured glomerular filtration rate in both heart and lung transplant patients. Despite reductions of more than 50% in CNI exposure, there was no marked loss of efficacy. The safety profile of the everolimus-based regimen was acceptable.

Cyclosporine A (CsA) has been shown to protect against myocardial ischemia and reperfusion (I/R) injury in small animal models. The aim of the current study was to evaluate the effects of CsA on myocardial I/R injury in a porcine model. Pigs were randomized between CsA (10mg/kg; n = 12) or placebo (n = 15) and anesthetized with either isoflurane (phase I) or pentobarbital (phase II). By catheterization, the left descending coronary artery was occluded for 45 minutes, followed by reperfusion for 2 hours. Hearts were stained to quantify area at risk (AAR) and infarct size (IS). Myocardial biopsies were obtained for terminal dUTP nick end labeling and immunoblot analysis of proapoptotic proteins (apoptosis-inducing factor [AIF], BCL2/adenovirus E1B 19-kd interacting protein 3 [BNIP-3], and active caspase-3). Cyclosporine A did not reduce IS/AAR compared with placebo (49% vs 41%, respectively; P = .21). Pigs anesthetized with isoflurane had lower IS/AAR than pigs anesthetized with pentobarbital (39% vs 51%, respectively; P = .03). This reduction in IS/AAR seemed to be attenuated by CsA. Apoptosis-inducing factor protein expression was higher after CsA administration than after placebo (P = .02). Thus, CsA did not protect against I/R injury in this porcine model. The data suggest a possible deleterious interaction of CsA and isoflurane., Funding agencies: The study wasfinanced by grants from the Swedish state under the agreementbetween the Swedish government and the county councilsconcerning economic support research and education of doctors(ALF-agreement) and the Swedish Heart Lung Foundation.

One of the mechanisms for generation of tolerance involves immature dendritic cells (DCs) and a subpopulation of regulatory CD4+ CD25+ T lymphocytes (T REG). The purpose of this work was to analyze how Cyclosporine A (CsA), a widely used immunosuppressive drug, may affect T REG proliferation. Purified and activated murine DCs obtained from bone marrow precursors differentiated with rGMCSF were co-cultured with purified CFSE-labeled T REG from OTII mice, and their phenotype and proliferation analyzed by flow cytometry. Our data indicate that DCs differentiated in the presence of CsA show an altered phenotype, with a lower expression of MHC-II and a lower activating capacity. Additionally, these CsA-treated DCs show decreased production of IL-2 and IL-12 and increased IL-10 secretion when stimulated with LPS, indicating an effect on the polarization of the immune response. Interestingly, CsA-treated DCs show an anti-tolerogenic effect since they reduce the proliferation of T REG cells from 72 to 47%. Further inhibition to a 24% of T REG proliferation was obtained as a direct effect of CsA on T REG. In conclusion, the anti-tolerogenic effect of CsA should be considered in the planning of immunosuppression in the context of clinical transplantation.

Jsou stanoveny termodynamické disociační konstanty imunosupresiv a cytostatik azathioprinu, methotrexátu, rasagilinu báze, rasagilinu mesylátu a cyklosporinu A (IVAX Pharmacueticals s.r.o., Opava) technikou pH-spektrofotometrické titrace při 25°C a 37°C. Po stanovení počtu ve spektrech indikovatelných světlo-absorbujících částic algoritmem INDICES byla ze závislosti smíšených disociačních konstant na odmocnině z iontové síly nelineární regresní analýzou zkráceného Debye-Hückelova vztahu programem ADSTAT odhadnuta termodynamická disociační konstanta. Nalezené odhady byly v dobré shodě s predikovanými odhady strukturní analýzy kvantově-chemického programu PALLAS., Thermodynamic dissociation constants of cytostatics and immunosuppressants azathioprine, methotrexate, rasagiline base, rasagiline mesylate and cyclosporine A (IVAX Pharmacueticals s.r.o.) were estimated by technique of pH-spectrophotometric titration at 25°C and 3°C. The first step was the determination in the spectrum indicated particles by the algorithm INDICES, then was from a dependence of dissociation constants on the root of the ionic strength by non-linear regression analysis of Dębye ? Hückelov law with the programme ADSTAT estimated the thermodynamic dissociation constant. Found estimates of pKaT were in good agreement with predicted estimates of structural analysis quantum-chemical programme PALLAS., Katedra analytické chemie, Dokončená práce s úspěšnou obhajobou

Oxidative stress is known to play a key role in the progression of liver disease, including non-alcoholic steatohepatitis (NASH), which is often accompanied by hyperglycemia. This study examined the influence of high glucose on oxidative stress-induced hepatic cell death. Hc cells, a normal human hepatocyte-derived cell line, were cultured in normal-to-high glucose (5.5-22 mM)-containing medium with varying concentrations (0.01-1 mM) of hydrogen peroxide. In certain experiments, cyclosporine A (CyA), which inhibits the mitochondrial permeability transition (MPT) pore, or Z-VAD-FMK (z-VAD), a pan-caspase inhibitor, were added to the medium. Cell viability was evaluated using a colorimetric assay. The mode of cell death was determined by nuclear staining methods using Hoechst 33258 and Sytox green. Neither high glucose (22 mM) nor 0.05-0.5 mM of hydrogen peroxide alone killed Hc cells. However, a combination of the two induced cell death, causing the nuclei of Hc cells to become expanded rather than condensed, and the nuclear membrane to become weak. CyA, but not z-VAD, blocked cell death. These results suggest that a high glucose condition may cause human hepatocytes to undergo hydrogen peroxide-induced necrotic cell death., Molecular Medicine Reports, 1(3), pp.379-385; 2008

TEN and HUS are challenging complications with excessive mortality after HSCT. We report the development of these two conditions in combination in a nine-yr-old boy after HSCT from an unrelated donor. TEN with skin detachment of more than 90% of body surface area developed after initial treatment for GvHD. Within a few days of admission to the burns unit, the patient developed severe hemolysis, hypertension, thrombocytopenia, and acute renal failure consistent with HUS, apparently caused by CSA. The management included intensive care in a burns unit, accelerated drug removal using plasmapheresis, and a dedicated multi-disciplinary team approach to balance immunosuppression and infections management in a situation with extensive skin detachment. The patient survived and recovered renal function but requires continued treatment for severe GvHD. Suspecting and identifying causative drugs together with meticulous supportive care in the burns unit is essential in the management of these patients and long-term survival is possible.

The aberrant activation of T lymphocyte proliferation is one of the key events in organ transplant recipients and autoimmune disorders. The present study adopted a gel-based proteomics approach to define the proteins representative of the T cell proliferation and to discover the molecules that play critical roles during the suppression of T cell proliferation. Human T lymphocytes were isolated from healthy donors and primed with phytohemagglutinin (PHA) to undergo proliferation. Two medical fungal products with specific T cell activation inhibitory properties, cyclosporine A (CsA) and polysaccharopeptide (PSP), were used to study the proteins that manipulate T cell proliferation. After demonstrating their similar effects on cell proliferation, cell survival and interleuklin-2 (IL-2) secretion, significant quantitative protein alterations were detected between the CsA- and PSP-treated T cell proteome. These altered proteins were identified by MALDI-TOF and classified into 3 categories: (i) proteins affected by both CsA and PSP, (ii) proteins affected by CsA alone, and (iii) proteins affected by PSP alone. Most of these altered proteins have functional significance in protein degradation, the antioxidant pathway, energy metabolism and immune cell regulation.

The aberrant activation of T lymphocyte proliferation is one of the key events in organ transplant recipients and autoimmune disorders. The present study adopted a gel-based proteomics approach to define the proteins representative of the T cell proliferation and to discover the molecules that play critical roles during the suppression of T cell proliferation. Human T lymphocytes were isolated from healthy donors and primed with phytohemagglutinin (PHA) to undergo proliferation. Two medical fungal products with specific T cell activation inhibitory properties, cyclosporine A (CsA) and polysaccharopeptide (PSP), were used to study the proteins that manipulate T cell proliferation. After demonstrating their similar effects on cell proliferation, cell survival and interleuklin-2 (IL-2) secretion, significant quantitative protein alterations were detected between the CsA- and PSP-treated T cell proteome. These altered proteins were identified by MALDI-TOF and classified into 3 categories: (i) proteins affected by both CsA and PSP, (ii) proteins affected by CsA alone, and (iii) proteins affected by PSP alone. Most of these altered proteins have functional significance in protein degradation, the antioxidant pathway, energy metabolism and immune cell regulation.

The present study evaluated the combination of antithymocyte globulin (ATG) and cyclosporine A (CsA) in patients with low-risk myelodysplastic syndromes. Twenty patients (17 with refractory anemia and 3 with refractory anemia with excess blasts) received treatment with rabbit-ATG plus CsA. The overall response rate was 30% (6/20); three of the six responders had a complete response. The responses lasted 2-58 months, with two patients still being in complete remission at 42 and 58 months. Short-lasting cytogenetic remissions were achieved in two patients. ATG was poorly tolerated in patients over 70 years of age. Four out of 20 patients progressed to acute myeloid leukemia within a year. We conclude that immunosuppressive treatment may be a therapeutic option for selected patients with myelodysplastic syndrome.

Introduction. Encephalitozoon intestinalis, a parasite belonging to the phylum Microsporidia, is causes gastrointestinal infections in the immunocompromised host. A suitable pharmacologically immunosuppressed animal model for the study of natural E. intestinalis infection, which can establish the immune components that respond to this parasite, is lacking.Objective. To evaluate the effect of immunosuuppression with Cyclosporine A (CsA) in C57BL/ 6 mice on experimental infection with E. intestinalis infection.Materials And Methods. Eighty C57BL/6 mice were distributed in four treatment groups: Control, CsA-immunosuppressed mice without infection, immunocompetent and immunossuppressed mice infected with E. intestinalis. Mice were immunosuppressed with a weekly dose of 50 mg/Kg body weight of CsA, during the course of the study. Five mice from each group were sacrificed 2, 3, 4 and 6 weeks post-infection, to obtain blood for antibody testing and stool samples were analyzed to assess excretion of spores.Results. Production of specific IgG antibodies was significantly higher in the immunocompetent group as compared to the immunosuppressed group of experimentally infected mice. In the infected mice, parasites were not observed in any tissues different from the small intestine. However, spore excretion through the stool and duodenal liquid was higher in the group of immunosuppresed infected mice.Conclusion. Immunosuppression induced with CsA in the murine model did not allow parasite dissemination and illness progression, but raised kinetics of spore excretion and decreased the production of IgG antibodies., Introducción. Encephalitozoon intestinalis es un microsporidio parásito del intestino, que puede diseminarse en pacientes inmunocomprometidos. Existen referencias de modelos animales inmunosuprimidos para el estudio de la microsporidiosis utilizando fármacos que producen supresión total de la respuesta inmune; sin embargo, no se han estudiado los efectos de inmunosupresores con acción selectiva sobre los componentes de esta respuesta. Objetivo. Evaluar el efecto de la inmunosupresión con ciclosporina A (CsA) en ratones C57BL/6 infectados con E. intestinalis. Materiales y métodos. Se utilizaron 80 ratones C57BL/6 distribuidos en cuatro grupos: infectados, inmunosuprimidos e infectados, inmunosuprimidos no infectados y controles. La inmunosupresión con CsA (50 mg/kg) se realizó vía intraperitoneal durante todo el estudio. En la semanas 2, 3, 4 y 6 posteriores a la infección se obtuvo sangre para determinar los anticuerpos, y materia fecal para evaluar la cinética de excreción de esporas. Además, se extrajeron varios órganos para estudiar la histopatología y observar la posible diseminación del parásito. Resultados. La producción de anticuerpos IgG fue mayor en los ratones inmunocompetentes infectados que en los inmunosuprimidos infectados con E. intestinalis. No se encontró el parásito en órganos diferentes al intestino delgado en los dos grupos infectados. Sin embargo, la excreción de esporas, tanto en heces como en líquido duodenal, fue mayor en el grupo inmunosuprimido infectado. Conclusión. La CsA en el modelo en ratón no indujo la diseminación de E. intestinalis ni la exacerbación de la enfermedad, pero contribuyó al aumento en la cinética de excreción de esporas y la disminución de la producción de anticuerpos IgG en los ratones inmunosuprimidos infectados.

Adjuvant treatment with adriamycin has been suggested to improve results after liver transplantation for hepatocellular cancer. Here we have applied an animal model for evaluation of treatment with adriamycin and/or cyclosporine A on liver tumour growth. Three chemically induced rat liver tumours with various degree of differentiation were transferred to the spleens of syngenic rats. Each recipient group was divided into four subgroups, treated with adriamycin and/or cyclosporine A or none of the drugs. When the tumour was well differentiated no proliferation was found in any of the subgroups. When the tumour exhibited a more pronounced dysplasia, adriamycin stimulated tumour growth. This effect was further increased by cyclosporine. In the animals transplanted with the most aggressive tumour, adriamycin inhibited tumour growth. When given together with cyclosporine this inhibition was counteracted. These data suggest that adriamycin, especially when given together with cyclosporine, may have a stimulatory effect on liver tumour cell growth.

Background : The prognosis of liver transplantation for liver cancer is determined by recurrence in the liver graft. In this study, the effects of immunosuppressors, FK506 and cyclosporine A (CsA) on the migration of liver cancer cells were investigated. Methods : The effects of FK506 at concentrations of 1-100 ng/mL and CsA at 1-1000ng/mL on the growth of poorly and well differentiated human hepatocellular carcinoma cell lines, HLE and HuH-7, respectively, were examined. After treatment of these cells with FK506 and CsA, the growth of these cells, their cytotoxicities and invasion assay on the Matrigel basement membrane invasion chamber were evaluated. In addition, the effects of FK506 and CsA on the changes in the production of a soluble intercellular adhesionmolecule-1 (sICAM-1) of these cells were measured. Results : FK506 and CsA at concentrations of 1-10 ng/mL inhibited the growth of both HLE and HuH-7 and those immunosuppressors at concentrations over 100 ng/mL exhibited cytotoxicity on these cells. FK506 at concentration of 1ng/mL significantly inhibited the invasion of poorly differentiated HLE, but not well differentiated HuH-7, after treatment for 2-5 days in culture (p＜0.05), but FK 506 at 10 ng/mL showed less inhibitory efficient. CsA at concentrations of 1-10 ng/mL, however, did not inhibit or transiently inhibited the invasion of both cell lines. The production of ICAM-1 in HLE and HuH-7 was suppressed by FK506 at concentrations of 1-10 ng/mL after treatment for 3-5 days but the effect was not significant in the initial phase at days 1-2 and the last phase at days 5-6. Conclusions : FK506, but not CsA, at a clinical dose of 1ng/mL significantly inhibited the invasion of the poorly-differentiated HLE, but not HuH-7 and also inhibited the production of sICAM-1 in HLE.

We recently demonstrated that azodicarbonamide is an immunosuppressive compound that inhibits calcium mobilization in T lymphocytes. In this study, we show that azodicarbonamide prevents the progression of human CD4+ T lymphocytes into the G1 phase of the cell cycle, inhibits their blastogenesis, down-regulates their membrane expression of CD25 and CD69, and decreases their transcription of cytokine genes. Addition of the calcium ionophore A23187 completely restores T cell proliferation in the presence of azodicarbonamide. Furthermore, azodicarbonamide synergizes with cyclosporin A to inhibit CD4+ T cell proliferation. In conclusion, the immunosuppressive action of azodicarbonamide is mainly related to its effect on the calcium mobilization machinery and is synergistic with that of calcineurin inhibitors., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published

We recently demonstrated that azodicarbonamide is an immunosuppressive compound that inhibits calcium mobilization in T lymphocytes. In this study, we show that azodicarbonamide prevents the progression of human CD4+ T lymphocytes into the G1 phase of the cell cycle, inhibits their blastogenesis, down-regulates their membrane expression of CD25 and CD69, and decreases their transcription of cytokine genes. Addition of the calcium ionophore A23187 completely restores T cell proliferation in the presence of azodicarbonamide. Furthermore, azodicarbonamide synergizes with cyclosporin A to inhibit CD4+ T cell proliferation. In conclusion, the immunosuppressive action of azodicarbonamide is mainly related to its effect on the calcium mobilization machinery and is synergistic with that of calcineurin inhibitors., Journal Article, Research Support, Non-U.S. Gov't, info:eu-repo/semantics/published

Renal transplantation is presently the renal replacement therapy ol choice lor most patients suffering lrom end-stage renal lailure, mainly because ol the gain in quality ol lile after successlul allografting. During the past decade the results ol renal transplantation, in terms ol incidence ol rejection episodes and graft loss because ol acute rejection, have improved substantially due to the introduction ol cyclosporine A. However, it appears that, despite the decrease in morbidity rates lollowing renal transplantation, in the long run the improvement in patient survival is rather limited (1).The success in terms ol graft survival wilhout a parallel improvement in patient survival, also emphasizes th at comorbid conditions such as cardiovascular disease, become increasingly important lor the longterm prospeets ol this patient group.

Partial tumoral responses have been described after cyclosporin A treatment in refractory B-chronic lymphocytic leukemia patients. We report the effect of this therapy in five patients prospectively treated with oral conventional dose of cyclosporin A. Antitumoral efficiency was marginal (one sustained partial response among five patients) which, added to a potential worsening of natural and therapy-related immunodepression, limits its use in this instance.

SCOPUS: ar.j, info:eu-repo/semantics/published

type:text, The pharmacokinetic behavior of cyclosporine A (CyA), known as a potential immunosuppressive agent to prevent graft rejection in transplantation, was studied in patients with acute hepatitis and primary biliary cirrhosis (PBC). The ratios of blood concentration of total CyA (CyA and its metabolites), CyA, and CyA metabolites to dose/kg body weight, (t-CyA/dose, CyA/dose, and CyA-Met/dose, respectively) were significantly higher in patients with hepatitis than those in renal transplantation. In PBC patients these ratios showed a tendency to be smaller than those in renal transplantation, but were not significant. The ratio of CyA-Met/ CyA was higher in the patients with hepatitis and PBC than that in renal transplantation. It was highest in the patients with PBC. The ratio of CyA-Met/CyA was significantly increased with a decrease of liver functions evaluated by serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and total serum bilirubin (t-Bil). These results indicate that hepatic function affects the pharmacokinetic behavior of CyA and the increased ratio of CyA-Met/dose could be caused by a possible increased efflux of metabolites into the blood circulation due to impaired bile excretion. These results also indicate the importance of therapeutic drug monitoring (TDM) in the use of CyA with patients with hepatic dysfunction.

The pharmacokinetic behavior of cyclosporine A (CyA), known as a potential immunosuppressive agent to prevent graft rejection in transplantation, was studied in patients with acute hepatitis and primary biliary cirrhosis (PBC). The ratios of blood concentration of total CyA (CyA and its metabolites), CyA, and CyA metabolites to dose/kg body weight, (t-CyA/dose, CyA/dose, and CyA-Met/dose, respectively) were significantly higher in patients with hepatitis than those in renal transplantation. In PBC patients these ratios showed a tendency to be smaller than those in renal transplantation, but were not significant. The ratio of CyA-Met/ CyA was higher in the patients with hepatitis and PBC than that in renal transplantation. It was highest in the patients with PBC. The ratio of CyA-Met/CyA was significantly increased with a decrease of liver functions evaluated by serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and total serum bilirubin (t-Bil). These results indicate that hepatic function affects the pharmacokinetic behavior of CyA and the increased ratio of CyA-Met/dose could be caused by a possible increased efflux of metabolites into the blood circulation due to impaired bile excretion. These results also indicate the importance of therapeutic drug monitoring (TDM) in the use of CyA with patients with hepatic dysfunction.

The pharmacokinetic behavior of cyclosporine A (CyA), known as a potential immunosuppressive agent to prevent graft rejection in transplantation, was studied in patients with acute hepatitis and primary biliary cirrhosis (PBC). The ratios of blood concentration of total CyA (CyA and its metabolites), CyA, and CyA metabolites to dose/kg body weight, (t-CyA/dose, CyA/dose, and CyA-Met/dose, respectively) were significantly higher in patients with hepatitis than those in renal transplantation. In PBC patients these ratios showed a tendency to be smaller than those in renal transplantation, but were not significant. The ratio of CyA-Met/ CyA was higher in the patients with hepatitis and PBC than that in renal transplantation. It was highest in the patients with PBC. The ratio of CyA-Met/CyA was significantly increased with a decrease of liver functions evaluated by serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and total serum bilirubin (t-Bil). These results indicate that hepatic function affects the pharmacokinetic behavior of CyA and the increased ratio of CyA-Met/dose could be caused by a possible increased efflux of metabolites into the blood circulation due to impaired bile excretion. These results also indicate the importance of therapeutic drug monitoring (TDM) in the use of CyA with patients with hepatic dysfunction.

SCOPUS: ar.j, info:eu-repo/semantics/published

The pharmacokinetic behavior of cyclosporine A (CyA), known as a potential immunosuppressive agent to prevent graft rejection in transplantation, was studied in patients with acute hepatitis and primary biliary cirrhosis (PBC). The ratios of blood concentration of total CyA (CyA and its metabolites), CyA, and CyA metabolites to dose/kg body weight, (t-CyA/dose, CyA/dose, and CyA-Met/dose, respectively) were significantly higher in patients with hepatitis than those in renal transplantation. In PBC patients these ratios showed a tendency to be smaller than those in renal transplantation, but were not significant. The ratio of CyA-Met/ CyA was higher in the patients with hepatitis and PBC than that in renal transplantation. It was highest in the patients with PBC. The ratio of CyA-Met/CyA was significantly increased with a decrease of liver functions evaluated by serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and total serum bilirubin (t-Bil). These results indicate that hepatic function affects the pharmacokinetic behavior of CyA and the increased ratio of CyA-Met/dose could be caused by a possible increased efflux of metabolites into the blood circulation due to impaired bile excretion. These results also indicate the importance of therapeutic drug monitoring (TDM) in the use of CyA with patients with hepatic dysfunction.

Malignant catarrhal fever (MCF) is caused by two closely associated gamma herpes viruses namely alcelaphine herpesvirus 1 (AlHV-1) and ovine herpesvirus 2 (OvHV-2) and characterised with lymphocyte infiltration in non-lymphoid tissues, vasculitis and epithelial damage. The mechanism by which the viruses cause the disease is not fully understood. The hypothesis of this project was that MCF is initiated by aberrant gene expression in endothelium, epithelium and infected T cells of susceptible animals, because they are not the natural hosts for the viruses and the viruses will not have evolved in them. The first goal was to examine whether rabbit epithelium and bovine endothelium can be infected in vitro and in vivo with AlHV-1 using q PCR and, if infected whether viral transcripts could be identified in these tissue cells using q PCR and in situ hybridisation (ISH). The results revealed that endothelium and epithelium can be infected and latent infection can be established in them. This suggests the likelihood of establishing a similar type of infection in vivo. Secondly, the trial to identify latency-associated transcripts using 5-azacitidine treatment on bovine turbinate fibroblast (BT) cells and rabbit large granular lymphocytes (LGLs) was only partially successful. However, pan T antigen was expressed in 5-azacitidine treated but not untreated LGLs cells. This may indicate a function of the drug either directly or through the latency state. Transcriptome analysis in the infected and treated LGLs and BT cells showed that several pathways were affected by 5-aza although a possible latency (low transcript levels) was only seen in the BTs. Transcriptome analysis revealed similar pathways to those described for MCF in the tissues in vivo, and an effect of 5-aza on these. Viral transcripts analysis showed that genes related to productive/lytic cycles were higher than latent ones on day 17 of the in vivo experiment demonstrating that the virus may replicate at this stage

Malignant catarrhal fever (MCF) is caused by two closely associated gamma herpes viruses namely alcelaphine herpesvirus 1 (AlHV-1) and ovine herpesvirus 2 (OvHV-2) and characterised with lymphocyte infiltration in non-lymphoid tissues, vasculitis and epithelial damage. The mechanism by which the viruses cause the disease is not fully understood. The hypothesis of this project was that MCF is initiated by aberrant gene expression in endothelium, epithelium and infected T cells of susceptible animals, because they are not the natural hosts for the viruses and the viruses will not have evolved in them. The first goal was to examine whether rabbit epithelium and bovine endothelium can be infected in vitro and in vivo with AlHV-1 using q PCR and, if infected whether viral transcripts could be identified in these tissue cells using q PCR and in situ hybridisation (ISH). The results revealed that endothelium and epithelium can be infected and latent infection can be established in them. This suggests the likelihood of establishing a similar type of infection in vivo. Secondly, the trial to identify latency-associated transcripts using 5-azacitidine treatment on bovine turbinate fibroblast (BT) cells and rabbit large granular lymphocytes (LGLs) was only partially successful. However, pan T antigen was expressed in 5-azacitidine treated but not untreated LGLs cells. This may indicate a function of the drug either directly or through the latency state. Transcriptome analysis in the infected and treated LGLs and BT cells showed that several pathways were affected by 5-aza although a possible latency (low transcript levels) was only seen in the BTs. Transcriptome analysis revealed similar pathways to those described for MCF in the tissues in vivo, and an effect of 5-aza on these. Viral transcripts analysis showed that genes related to productive/lytic cycles were higher than latent ones on day 17 of the in vivo experiment demonstrating that the virus may replicate at this stage