CaRMeN: a tool for analysing and deriving kinetics in the real world.

This paper presents the concepts of an open software tool (CaRMeN) that can be used to rapidly analyse and derive models, in particular chemical kinetics. The software automates the workflow of comparing model vs. experiment, which must currently be done manually and is thus a time-consuming and error-prone task. The capabilities of the software are illustrated through a case study. Experimental data for the conversion of methane over rhodium catalysts in a wide range of conditions and experimental setups are numerically simulated using five different mechanisms from the literature. The applicability of the mechanisms as well as differences between flow and diffusion models are evaluated. The results show that no single mechanism reliably predicts the chemical conversions of all of the experiments. Although the software was initially developed for chemical kinetics applications, it can also be extended to run any simulation code, and can therefore be applied in other scenarios.

Good for the lung but bad for the liver? Garlic-induced hepatotoxicity following liver transplantation.

WHAT IS KNOWN AND OBJECTIVE: Limited data exist surrounding the metabolism and safety of garlic supplements. CASE DESCRIPTION: A patient with a history of hepatopulmonary syndrome (HPS) and orthotopic liver transplantation was admitted to our surgery transplant service with severe hypoxaemia. The patient was started on high-dose Garlicin Cardio® (Allium sativum) for HPS and soon after had elevated liver function tests. Garlicin Cardio® was discontinued and liver enzymes normalized. A liver biopsy revealed mild periportal cholestatic reaction suggesting potential drug-induced aetiology. WHAT IS NEW AND CONCLUSION: This is the first description of liver injury secondary to garlic supplementation. Therefore, this garlic supplement should be listed as a potential cause of acute drug-induced liver injury.

Symptoms, diagnoses, and sporting consequences among athletes referred to a Danish sports cardiology clinic.

As the number of recreational athletes performing exercise and participating in competitions at a high-level increases, exercise-induced cardiac symptoms may become a more common problem, not least because recreational athletes often continue high-level exercise programs into advanced ages. We investigated the prevalence of cardiac symptoms and diagnoses among 201 athletes referred for cardiac evaluation at a Sports Cardiology Clinic in Denmark. To our knowledge, this is the first systematic study of athletes referred for suspected cardiac disease. The athletes were all well-trained recreational to elite athletes who participated in various sports with different training loads and a wide age span (13-66 years). All patients were referred by physicians, primarily their general practitioner (38%), and palpitations were the most common cardiac symptom (40%). Cardiac symptoms had a sensitivity of 86% in detecting cardiac disease and a specificity of 13%. Cardiac disease was diagnosed in 44% of the patients, and atrial fibrillation was the most prevalent diagnosis (7.5%). Cardiac diseases with therapeutic- or sports-related consequences for the patients were diagnosed in 28% of the population, but only 1% received a recommendation to avoid high-level sports indefinitely.

Monitoring of adenovirus (ADV)-specific T cells in a boy with ADV pneumonia and disseminated disease after lung transplantation.

Human adenovirus (ADV) infections are the cause of severe morbidity and mortality in transplant recipients. Cidofovir (CDV) is the current standard antiviral treatment. We report the case of a 3-year-old boy after lung transplantation with severe ADV sepsis, who was monitored for ADV-specific T cells during his disease and recovery. A strong increase in ADV-specific T cells was accompanied by resolution of ADV in blood and bronchoalveolar lavage fluid. Antiviral treatment with CDV was individually adapted according to anti-ADV immune responses, which provides a new method for tailoring antiviral treatment in lung transplant recipients.

Granulocyte colony-stimulating factor impairs CD8(+) T cell functionality by interfering with central activation elements.

Besides mobilizing stem cells into the periphery, granulocyte colony-stimulating factor (G-CSF) has been shown to influence various types of innate and adaptive immune cells. For example, it impairs the effector function of cytotoxic T lymphocytes (CTLs). It is assumed that this effect is mediated indirectly by monocytes, regulatory T cells and immunomodulatory cytokines influenced by G-CSF. In this study, isolated G-CSF-treated CD8(+) T cells were stimulated antigen-dependently with peptide-major histocompatibility complex (pMHC)-coupled artificial antigen-presenting cells (aAPCs) or stimulated antigen-independently with anti-CD3/CD28 stimulator beads. By measuring the changes in interferon (IFN)-γ and granzyme B expression at the mRNA and protein level, we showed for the first time that G-CSF has a direct effect on CD8(+) CTLs, which was confirmed based on the reduced production of IFN-γ and granzyme B by the cytotoxic T cell line TALL-104 after G-CSF treatment. By investigating further elements affected by G-CSF in CTLs from stem cell donors and untreated controls, we found a decreased phosphorylation of extracellular-regulated kinase (ERK)1/2, lymphocyte-specific protein tyrosine kinase (Lck) and CD3ζ after G-CSF treatment. Additionally, miRNA-155 and activation marker expression levels were reduced. In summary, our results show that G-CSF directly influences the effector function of cytotoxic CD8(+) T cells and affects various elements of T cell activation.

Osteoporosis in the adult solid organ transplant population: underlying mechanisms and available treatment options.

The prevention and treatment of osteoporosis is an increasingly important topic in the solid organ transplant (SOT) population. Compared to the general population, these patients are at an elevated risk of developing osteoporosis due to progressive disease, lifelong immunosuppressant therapy, and malnutrition. As patients live longer after transplant, chronic disease management is increasingly more important. Supplementation with calcium and vitamin D is often necessary in the SOT population due to a high incidence of vitamin D deficiency. Bisphosphonate therapy is most commonly used for prevention and treatment of osteoporosis, but therapy can be limited by renal dysfunction which is common in transplant recipients. Alternative agents such as teriparatide and calcitonin have not been shown to provide a significant impact on the rate of fractures in this population. Additionally, denosumab may be a promising treatment option due to its novel mechanism of action, and is currently being studied in renal transplant patients. Timely initiation of supplementation and treatment, and minimizing glucocorticoid exposure prior to and after transplantation will aid in the prevention and proper management of osteoporosis in these patients.

Results of voluntary cardiovascular examination of elite athletes in Denmark: Proposal for Nordic collaboration.

We investigated the cardiovascular status of elite athletes in Denmark, the extent of abnormal cardiac findings--both training related and pathologic--and how participating in cardiac examination was perceived by the athletes. A standardized protocol of questionnaires, physical examination, resting electrocardiogram, and 2D echocardiography was used. In total 1347 elite athletes were invited; 516 athletes (38%) from 30 different sports participated. Results were stored in a web-based database for future research and long-term follow-up. Cardiac pathology was infrequent; eight athletes (1.6%) received a cardiac diagnosis; one athlete (0.2%) diagnosed with long QT syndrome was advised against competition level sports. In total, 60 athletes (11.6%) were referred for additional testing. The athletes presented a very low level of psychological stress before and a slight decrease immediately after the examination as measured by the REST-Q 76 Sport questionnaire. Athletes needing further examinations did not present a higher level of stress after the initial examination compared with athletes with normal test results. Overall, very few athletes were diagnosed with a cardiac condition that increased risk of sudden cardiac death. Less than half of the invited athletes volunteered, but participation was not perceived stressful by the enrolled athletes, not even when additional testing was needed.

Modulation of heme oxygenase-1 by metalloporphyrins increases anti-viral T cell responses.

Heme oxygenase (HO)-1, the inducible isoform of HO, has immunomodulatory functions and is considered a target for therapeutic interventions. In the present study, we investigated whether modulation of HO-1 might have regulatory effects on in-vitro T cell activation. The study examined whether: (i) HO-1 induction by cobalt-protoporphyrin (CoPP) or inhibition by tin-mesoporphyrin (SnMP) can affect expansion and function of virus-specific T cells, (ii) HO-1 modulation might have a functional effect on other cell populations mediating effects on proliferating T cells [e.g. dendritic cells (DCs), regulatory T cells (T(regs)) and natural killer cells] and (iii) HO-1-modulated anti-viral T cells might be suitable for adoptive immunotherapy. Inhibition of HO-1 via SnMP in cytomegalovirus (CMV)pp65-peptide-pulsed peripheral blood mononuclear cells (PBMCs) led to increased anti-viral T cell activation and the generation of a higher proportion of effector memory T cells (CD45RA(-) CD62L(-)) with increased capability to secrete interferon (IFN)-γ and granzyme B. T(reg) depletion and SnMP exposure increased the number of anti-viral T cells 15-fold. To test the possibility that HO-1 modulation might be clinically applicable in conformity with good manufacturing practice (GMP), SnMP was tested in isolated anti-viral T cells using the cytokine secretion assay. Compared to control, SnMP treatment resulted in higher cell counts and purity without negative impact on quality and effector function [CD107a, IFN-γ and tumour necrosis factor (TNF)-α levels were stable]. These results suggest an important role of HO-1 in the modulation of adaptive immune responses. HO-1 inhibition resulted in markedly more effective generation of functionally active T cells suitable for adoptive T cell therapy.

The new HLA-B*58:21 allele is predicted to be functionally similar to the B*58:01P group of alleles.

The new human leukocyte antigen (HLA)-B*58:21 allele differs from B*58:01:01 by an amino acid exchange at codon 90.

Early aspirin therapy may reduce hepatic artery thrombosis in liver transplantation.

BACKGROUND: Hepatic artery thrombosis (HAT) remains among the leading causes of early graft loss after liver transplantation. Our transplant center began using universal aspirin prophylactic therapy immediately posttransplantation in 2007. The aim of this study was to determine the safety and efficacy of early aspirin therapy on clinical outcomes. METHODS: This large-scale, cross-sectional analysis included all adult liver transplantations performed between 2000 and 2009. Pediatric and multiorgan transplants were excluded. Patients were grouped and compared based on whether they received early initiation of aspirin 325 mg PO daily posttransplantation. RESULTS: A total of 541 adult liver transplantations occurred during the study period; 439 had complete documentation and were analyzed. Clinical outcomes show aspirin patients had similar rates of early and late HAT, but had significantly lower early HAT, defined as HAT occurring within the first 30 days posttransplant, leading to graft loss. Other clinical outcomes were similar between groups including bleeding events and wound complications. CONCLUSIONS: Immediate initiation of aspirin therapy after liver transplantation may reduce the rate of HAT leading to early graft loss, without increasing bleeding or other complication rates.

The impact of early corticosteroid withdrawal on graft survival in liver transplant recipients.

BACKGROUND: There has been increased interest in recent years in reducing or eliminating steroids from the immunosuppression regimen of transplant recipients to reduce adverse effects associated with their use. The purpose of this study was to compare clinical outcomes between early versus late steroid withdrawal after liver transplant to determine the optimal duration of steroid use in this population. METHODS: This large-scale, retrospective analysis of liver transplants occurred at our institution between 2000 and 2009. Patients were excluded if they were <18 years old, received a multiorgan transplant, or remained on steroids for >1 year. The early steroid withdrawal group had steroids eliminated by 3 months posttransplant; late steroid withdrawal patients had steroids withdrawn between 3 and 12 months posttransplant. RESULTS: A total of 586 liver transplants occurred during the study period; 330 patients were included in the analysis. Graft survival was significantly lower in the early steroid withdrawal group. There was no difference in patient survival or overall acute rejection. However, the late steroid withdrawal group had a significantly higher rate of early acute rejection episodes. There was no difference with regard to new-onset diabetes after transplant, hyperlipidemia, or cardiovascular events between groups. CONCLUSION: The results of this study suggest that late corticosteroid withdrawal is associated with better long-term graft survival without increasing the rates of diabetes, hyperlipidemia, or cardiovascular events in liver transplant recipients. A prospective study is warranted to confirm these findings.

Closing the gap: discrimination of the expression profile of HLA questionable alleles by a cytokine-induced secretion approach using HLA-A*32:11Q.

Matching of human leukocyte antigen (HLA) alleles between donors and recipients plays a major role in hematopoietic stem cell transplantation (HSCT). Null or questionably expressed HLA allelic variants are a major issue in HLA matching, because the aberrant expression of such alleles can have a major impact on the outcome of HSCT and/or its complications such as graft-versus-host disease. The goal of this study was to investigate the potential of a recently developed cytokine-induced secretion assay to differentiate the expression levels of HLA-A*32:11Q (questionable) into a null (N) or low (L) expression variant. An amino acid mutation at position 164 of HLA-A*32:11Q disrupts the disulfide bridge in the α2 domain. HLA-A*32:11Q is not detectable by standard microlymphocytotoxicity assay. To this end, we cloned soluble HLA-A*32:11Q and a reference allele (HLA-A*32:01) into expression vectors and transfected/transduced HEK293 and K562 cells. Allele-expressing K562 cells were simultaneously transfected/transduced with a ß2-microglobulin (B2M)-encoding vector to ensure the intact HLA structure with B2M. After treatment with proinflammatory cytokines, secreted soluble HLA molecules were determined by enzyme-linked immunosorbent assay in the supernatant and intracellular accumulation of the recombinant proteins by flow cytometry. HLA-A*32:11Q was nearly undetectable in untreated transfectants. Cytokine treatment increased the secretion of HLA-A*32:11Q to detectable levels and resulted in intracellular accumulation of the allele. There was no difference in mRNA transcription between the A*32 alleles. On the basis of these results, we recommend reclassification of HLA-A*32:11Q as a low expression (L) variant.

Modulatory role of calreticulin as chaperokine for dendritic cell-based immunotherapy.

Heat shock proteins (HSPs) play a regulatory role for maturation of antigen-presenting cells (APCs) such as dendritic cells (DCs) and macrophages. Whereas HSP70 has been shown to enhance the maturation of human DCs via a nuclear factor kappa-B (NF-κB)-dependent pathway, the regulatory role of calreticulin (CRT), which is a HSP with similar functions to HSP70, is not well studied. To investigate the role of CRT as adjuvant in cell activation and co-stimulatory responses we determined the effects of CRT on human APC maturation in comparison to that of HSP70. To facilitate eukaryotic endotoxin-free CRT protein expression, three different methods were compared. We demonstrate that CRT induces the maturation of human DCs and increases the production of proinflammatory cytokines via the NF-κB pathway. CRT-mediated maturation was qualitatively similar to that induced by HSP70. Interestingly, priming of monocytes with HSPs showed an even more prominent effect on maturation than exposure of immature DCs to these compounds. A higher expression of CD86, CD83 and CCR7 on mature DCs were found in response to CRT. Our data provide novel insights into the role of extracellular HSPs as chaperokines in the processes of APC generation and may thus be useful to improve adoptive immunotherapy.

The amino acid variation of HLA-A*02:182 in a highly conserved region is predicted to be functionally similar to HLA-A*02:01:01:01 allele.

We here describe the identification of the novel human leukocyte antigen allele HLA-A*02:182 which has been detected in a potential bone marrow donor. The new allele differs from the sequence of HLA-A*02:01:01:01 only by a non-synonymous nucleotide exchange of Guanin (G) → Cytosin (C) at position 199 in exon 3 replacing amino acid (AA) Arginine (Arg, R) by Threonine (Thr, T) in codon 157. Since the HLA-A*02:01:01:01 allele differs from A*02:182 only at AA position 157, it is assumed that the protein structures of these alleles are highly similar. A mismatch between HLA-A*02:01:01:01 and HLA-A*02:182 is predicted to have a very low allogeneic potential in hematopoietic stem cell transplantation.

The predicted protein structures of the novel DRB1*0717 and DRB1*0701 are highly related.

This article describes the identification of the novel human leukocyte antigen (HLA) allele DRB1*07 7 that was detected in a potential stem-cell donor of Caucasian origin. Compared to DRB1*070101, the new allele is characterized by a nonsynonymous nucleotide exchange of C-->T at position 201 in exon 2 replacing Arg by Trp in codon 72. As this sequence variation has not been seen earlier in any other HLA-DRB allele, it is most probably the result of a point mutation.

Plant--rhizosphere-microflora association during phytoremediation of PAH-contaminated soil.

The capability of plants to promote the microbial degradation of pollutants in rhizosphere soil is a principal mechanism of phytoremediation of PAH-contaminated soil. The formation of a specific rhizosphere microbocenosis with a high degradative potential toward contaminants is largely determined by plant species. The comparative PAH-degradation in unplanted soil and in soil planted with reed (Phragmites australis) and alfalfa (Medicago sativa) was studied in pot experiments during 2 years. Both alfalfa and reed successfully remediated contaminated soil by degrading 74.5 and 68.7% of PAHs, respectively. The study of the rhizosphere, rhizoplane, and unplanted-soil microflora in experimental pots showed that alfalfa stimulated the rhizosphere microflora of PAH-contaminated soil more effectively than did reed. Alfalfa clearly enhanced both the total number of microorganisms (1.3 times, according to fluorescence microscopy data) and the rate of the PAH-degrading population (almost seven times, according to plate counting). The degradative potential of its rhizosphere microflora toward PAHs was higher than the degradative activity of the reed rhizosphere. This study provides relevant information for the successful application of alfalfa to phytoremediate PAH-contaminated soil.

Making Connections Efficiently: A Comparison of Two Approaches Used by College Students to Construct Networks.

This study compared the effects of two approaches to networking. Two intact classes of college students participated in this study. The major focus of this study was to examine the extent to which the mental construction of networks was sufficient for recall to occur. Two intact classes of students participated in this study. Experimental participants were taught to construct their own networks both on paper and mentally while control participants were not. Recall of ideas was assessed at the pretest and at the posttest using a free-written recall task and a multiple-choice test. The results showed that networking mentally is as effective for recalling ideas and more time-efficient than networking on paper. Moreover, participants used different networking modalities depending on the passage to be read and the level of detail they are attempting to recall. The results are promising as they provide college students with a powerful tool that encourages cognitive processing without unnecessarily depleting their time resources. However, the results of this study also reveal that some initial experience with the strategy is necessary. Copyright 1999 Academic Press.