Altered hetero- and homeometric autoregulation in the terminally failing human heart

OBJECTIVE AND METHODS: To further investigate length-dependent force generation in human heart, nonfailing (donor hearts, NF) and terminally failing (heart transplants, dilated cardiomyopathy, DCM) left ventricular myocardium was studied under various preload (4-40 mN/mm2) or length conditions. In addition, morphological studies (van Giesson Trichrome staining, electron microscopy) were performed. RESULTS: In NF, a biphasic increase in force of contraction (FOC) was observed after elevating the preload (4-40 mN/mm2): there was an immediate fast increase (FOCf,), followed by a slow increase over several minutes (FOCs), which was paralleled by an increase in the systolic fura-2 transient. In DCM, FOCf, FOCs and the systolic fura-2 transient were blunted and diastolic tension was increased at increasing muscle length. Only in NF, a stretched induced increase in diastolic fura-2 ratio was observed. In DCM, no obvious interstitial fibrosis and no difference in basement membrane structure and attachment were observed. CONCLUSIONS: Since FOCf has been attributed to the Frank-Starling mechanism, whereas FOCs represents a length-dependent increase in the intracellular Ca2+-transient, the impaired length-dependent force generation in failing myocardium results from a dysregulation of both myofibrillar Ca2+-sensitivity as well as the intracellular Ca2+-homeostasis. Interstitial fibrosis may have only minor impact on force generation in human end-stage heart failure.     

Community-acquired methicillin-resistant Staphylococcus aureus outbreak in a local high school football team unsuccessful interventions

A retrospective review of an outbreak of community-acquired methicillin-resistant Staphylococcus aureus soft tissue infections in a high school football team was conducted. Thirteen players had 20 infections. Available community-acquired methicillin-resistant S. aureus isolates showed identical antibiotic susceptibility and field inversion gel electrophoresis analysis band patterns. Nasal cultures and mupirocin were ineffective at predicting and controlling infection, respectively. Infections treated with beta-lactam antibiotics were at risk for recurrence.     

The Use of an Hydrogen Peroxide Multipurpose Isolator for Inhouse Preparation of Human Embryo Culture Media: A Unique Successful Swiss Randomized Prospective Study

Purpose: To develop inhouse made (IHM) embryo culture medium with a Multipurpose Isolator and compare the embryo development in a prospective randomized study with commercial media.Methods: Fertilization by intracytoplasmic single sperm injection (ICSI) of Metaphase II oocytes obtained after 96 controlled ovarian hyperstimulation cycles in patients not older than 37 years. Transfer of zygotes to IHM or commercial Cook Sydney IVF Cleavage medium (SIC) immediately after pronucleus observation. Evaluation of embryo cleavage and score, pregnancy, and implantation rate.Results: From 100 zygotes cultured in SIC, 61% were at the 4 cell stage 45 h after ICSI compared to 77% (78/101) in the IHM, P<0.05. The mean embryo score with IHM was 3.9±0.9 compared to 3.5±1.2 with SIC, P<0.05. The clinical pregnancy rate per transfer was 38.9% (37/95), the implantation rate was 23% (46/200), and no differences were observed between the groups.     

A novel connexin 26 mutation associated with autosomal recessive sensorineural deafness

Mutations in the connexin 26 (Cx26) gene (GJB2) are a common cause of hereditary hearing impairment which affects approximately 1 in 2000 newborn children. We report the identification of a novel Cx26 point mutation (439 G-->A) linked to familial, autosomal recessive, sensorineural hearing loss. This missense mutation (E147K) is located in the highly conserved, putative K(+) channel lining sequence of the third transmembrane domain (TM3) of Cx26. Hearing impairment associated with this mutation was congenital, moderate to profound and showed no signs of progressive deterioration.     

Differential effects of rapamycin, cyclosporine A, and FK506 on human coronary artery smooth muscle cell proliferation and signalling

BACKGROUND: Immunosuppressive agents are at the forefront of preventing organ rejection after transplantation. However, their effects on vascular smooth muscle cell-mediated intimal hyperplasia that occurs in post-transplant coronary artery disease are less well known. METHODS AND RESULTS: We investigated the in vitro effects of three immunosuppressive agents cyclosporine A (CsA), FK506 (tacrolimus), and rapamycin (sirolimus, Rapa) on cultured human coronary artery smooth muscle cells (cSMC). CsA inhibited both platelet-derived growth factor (PDGF)-stimulated DNA synthesis and serum-induced proliferation at high concentrations (> or =1000 ng/ml). The growth-inhibitory effect of CsA was not altered by anti-TGF-beta neutralising antibodies nor was autocrine TGF-beta release detected in CsA-treated culture medium. At inhibitory doses, CsA inhibited ERK kinase activation by PDGF, although cytotoxicity was also apparent. Most notably, CsA visibly prevented PDGF-induced altered cell morphology. Rapa was a highly potent and effective inhibitor of cSMC proliferation (reduction in DNA synthesis by >50% from 0.01 ng/ml), acting through inhibition of 70-kDa S6 kinase (p70S6k). FK506 (1-1000 ng/ml) did not affect cSMC proliferation alone, although a > or =250-fold excess of FK506 over Rapa completely reversed the inhibitory effect of Rapa, confirming that these two agents share a common intracellular receptor, the FK506-binding protein (FKBP). CONCLUSION: Rapa is a powerful inhibitor of cSMC proliferation, while CsA slighly inhibits cSMC proliferation, although only at higher concentrations that may be toxic. These results indicate that therapeutic immunosuppression with Rapa may be additionally useful in prevention or delay of posttransplant coronary artery disease.     

CD95/Fas expression on peripheral blood T lymphocytes in patients with multiple sclerosis: effect of high-dose methylprednisolone therapy

Recent data indicate that the apoptotic process, mediated by the CD95/Fas cell surface receptor, is impaired in activated lymphocytes of patients with relapsing-remitting multiple sclerosis. Using flow cytometric-immunophenotyping, we analyzed the expression of CD95/Fas on peripheral blood CD4+ and CD8+ T lymphocytes (PBL) in 10 MS patients in relapse, and the effect of pulse corticosteroid therapy on the apoptosis of autoreactive lymphocytes. The proportions of CD8+ and CD8+CD95+ T lymphocytes were significantly higher in MS patients in relapse before than after pulse corticosteroid therapy. Conversely, the proportions of CD4+ and CD4+CD95+ T cells were significantly lower before than after therapy, but not significantly different from healthy persons. The different expression of CD95/Fas on peripheral blood CD8+ T lymphocytes in relapsing RRMS and in healthy controls suggests a possible involvement of apoptosis in the pathogenesis of MS. Our results also show that pulse corticosteroid therapy influences the CD95/Fas expression on CD8+ and CD4+ T lymphocytes in patients with RRMS.     

Induction of neutralising antibodies restricts the use of human granulocyte/macrophage colony stimulating factor for vaccine studies in rhesus macaques

Granulocyte/macrophage-colony stimulating factor (GM-CSF) is a valuable adjuvant to enhance induction of cellular immune responses in rodents. Less information is available regarding its use as an adjuvant in primates or humans. We explored recombinant human GM-CSF for potential vaccine studies in rhesus macaques and focused on its effect on peripheral monocytes as progenitors of dendritic cells and its potential immunogenicity. Application of human GM-CSF to nine animals led to an average 32-fold increase in monocyte numbers. This was not observed upon re-treatment, which coincided with GM-CSF-specific neutralising antibodies. These also neutralised the activity of rhesus macaque GM-CSF. The data underscore the need to use species-specific GM-CSF for immunomodulation in primates.     

Hidden financial costs in treatment for childhood cancer: an Australian study of lifestyle implications for families absorbing out-of-pocket expenses

PURPOSE: The impact of out-of-pocket expenses on five domains of family lifestyle were explored: social, assets, credit, utilities, and charity. METHODS: Using a cross-sectional survey, 100 parents of pediatric cancer patients reported on the types of out-of-pocket expenses incurred and the perceived lifestyle impact of meeting those expenses. RESULTS: Eighty percent of the sample reported a minimum of five different out-of-pocket expenses (total mean value = 19,064 Australian dollars; approximately 9,723 US dollars). The majority reflected travel, accommodation, and communication costs, use of work-related entitlements, and changes in paid employment. In lifestyle terms, the area of greatest impact was found for the social domain, such as cancelling vacations and giving up recreational pleasures and social expenditure. Those families living furthest from the major cancer treatment center reported the greatest range of out-of-pocket expenses and subsequent lifestyle impact. While there were few differences as a function of cancer type, results suggested that families most vulnerable to financial distress tended to be those whose child had spent relatively longer on treatment. CONCLUSIONS: In meeting out-of-pocket expenses, parents primarily seek ways to "trim the fat" off existing family expenditure. While all families may incur extra expenses, parents of patients located a significant distance from the cancer treatment center remain especially vulnerable (despite increased government allowances). Creative solutions for addressing some expenses may include applications of telemedicine to augment outreach services.     

PARC/CCL18 is a plasma CC chemokine with increased levels in childhood acute lymphoblastic leukemia

Chemokines play an important role in leukocyte mobilization, hematopoiesis, and angiogenesis. Tissue-specific expression of particular chemokines also influences tumor growth and metastasis. Here, the CC chemokine pulmonary and activation-regulated chemokine (PARC)/CCL18 was measured in pediatric patients with acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML). Surprisingly, PARC immunoreactivity was consistently detected in plasma from healthy donors. After purification to homogeneity, the presence of intact PARC (1-69) and processed PARC (1-68) in normal human plasma was confirmed by sequence and mass spectrometry analysis. Furthermore, PARC serum levels were significantly increased in children with T-ALL and prepreB-ALL compared to control serum samples, whereas serum levels in AML and preB-ALL patients were not significantly different from controls. In contrast, the hemofiltrate CC chemokine-1 (HCC-1)/CCL14 was not found to be a biomarker in any of these patients' strata, whereas the cytokine interleukin-6 (IL-6) was significantly decreased in AML and prepreB-ALL. Stimulated leukocytic cell lines or lymphoblasts from patients produced IL-8/CXCL8 or macrophage inflammatory protein-1alpha (MIP-1alpha/CCL3) but not PARC, not even after IL-4 or IL-10 treatment. However, PARC was produced by superantigen or IL-4 stimulated monocytes co-cultured with lymphocytes or lymphoblastic cells. Serum PARC levels thus constitute a novel leukemia marker, possibly reflecting tumor/host cell interactions in the circulation.