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Background Elevations of interleukin 6 (IL-6) have been described in drug-induced anaphylaxis. Although IL-6 is well known to stimulate an acute phase response, profiling acute phase protein levels, such as C-reactive protein (CRP), has, to our knowledge, never been performed in patients with acute allergic reactions. Objective To examine the pattern of IL-6 and CRP levels in patients with acute allergic reactions and to relate these to relevant clinical and laboratory parameters. Methods Plasma CRP and serum IL-6 levels were determined in 85 adult emergency department patients. These patients had been previously studied with questionnaires, physical examinations, and histamine/tryptase levels. Clinical and historical features were related to CRP and IL-6 levels. CRP and IL-6 levels were also examined for relationships with histamine and tryptase levels. Results CRP and IL-6 levels were significantly correlated with one another in the study patients (Spearman ρ = 0.36, P = 0.0008). Similar to histamine levels, IL-6 levels were significantly correlated with the extent of erythema manifested by the study patients. The extent of erythema was independently predicted by both IL-6 and histamine levels. Histamine levels were negatively correlated with CRP levels (Spearman ρ = −0.32, P = 0.003). Unlike histamine levels, IL-6 and CRP did not show significant relationships with the extent or presence of urticaria/angioedema or the presence of wheezing. IL-6 levels were correlated with the duration of symptoms before serologic sampling. An inverse correlation was observed between IL-6 levels and mean arterial blood pressure. Multivariate modeling showed significant independent effects from mean arterial pressure, duration of symptoms, erythema extent, and age in predicting IL-6 levels. Tryptase levels were higher in patients whose IL-6 levels were >20 pg/mL. Conclusions CRP and IL-6 levels are not simple surrogate markers for histamine or tryptase release by mast cells or basophils in acute allergic reactions. Increasing IL-6 levels relate to greater erythema extent, lower mean arterial blood pressure, and a longer duration of symptoms. It would be interesting to speculate that CRP and IL-6 increases characterize a late-phase response in immediate hypersensitivity reactions. In this perspective, the inverse relationship between CRP and histamine levels could be explained. As histamine levels are waning, CRP levels are increasing. Timed studies for histamine and CRP/IL-6 levels in allergic reactions are necessary to confirm this hypothesis. more...

Insulin-like growth factor-1 (IGF-1) is a highly mitogenic polypeptide detectable in human lung. Using competitive reverse transcriptase/polymerase chain reaction (RT-PCR), expression of four IGF-1 transcripts was examined in bronchoalveolar lavage cells (BALC) from normal subjects, idiopathic pulmonary fibrosis (IPF), stage I/II (no fibrosis), and stage III/IV (confirmed fibrosis) pulmonary sarcoidosis patients, and fibroblast strains isolated from normal and IPF lungs. Transcripts studied were Class 1 and Class 2 (exons 1 or 2, respectively) with IGF-1Eb or IGF-1Ea (exons 5 or 6, respectively). Total IGF-1 expression was downregulated in BALC of both patients with IPF (p0.01) and patients with sarcoidosis (p0.04) compared with healthy subjects. In contrast, both constitutive (p0.003) and transforming growth factor-beta (TGF-beta)- induced (p0.02) IGF-1 expression was higher in fibrotic, compared with normal, fibroblasts. These changes were associated with differential expression of IGF-1 splice variants. Healthy subjects and sarcoidosis patients without fibrosis showed similar expression of Class 1/Class 2 and IGF-1Ea/IGF-1Eb. However, patients with fibrosis demonstrated discordant, increased relative abundance of Class 1 transcripts (p0.01). In parallel, all fibrosis patients failed to express Class 2, IGF-1Eb forms and sarcoidosis patients with fibrosis did not express the Class 1, IGF-1Eb variant either. Fibrotic fibroblasts expressed higher constitutive levels of Class 1, IGF-1Ea transcripts compared with normal fibroblasts. Class 2, IGF-1Eb forms were moderately expressed by fibroblasts only after stimulation with TGF-beta, which also further increased levels of Class 1, IGF-1Ea transcripts. Our findings suggest that transition from a healthy to a fibrotic phenotype occurs in association with a changing pattern of IGF-1 mRNA heterogeneity and leads us to hypothesize a potential role for specific IGF-1 variants in fibrogenesis. more...

Corvid populations are increasing worldwide in response to urbanization. We investigated the response of American Crows (Corvus brachyrhynchos) to urbanization by (1) comparing rates of winter population change between urban and nonurban locations (using standard Christmas Bird Counts); (2) quantifying population size along a gradient of urbanization in western Washington; and (3) pooling studies from eastern (New York), midwestern (Wisconsin), and western North America (Washington and California) relating survivorship, reproduction, and space use to urbanization. American Crow populations tend to be densest and increasing most rapidly in urban areas of North America. This appears to be facilitated by small space neEDS of crows in urban relative to suburban, rural, and exurban areas. Crow survivorship is high across the urban gradient, but reproduction and hence population growth, peaks in suburban and rural settings. Local demographic considerations appear unable to account for changing winter crow populations. Rather, we hypothesize that urban crow populations may be increasing primarily as surplus crows from suburban and rural areas disperse into the city where anthropogenic food sources are easily located, rich, and concentrated. This hypothesis likely is affected by local crow sociality. In the western United States, where pre-breeders often form flocks able to exploit urban riches, our dispersal hypothesis may be accurate. But, in midwestern and eastern areas, where crows migrate south for winter or remain on territories to help rather than float as pre-breeders, dispersal may not be adequate to fuel urban population growth. Refuse, invertebrates, and small vertebrates appeared to be more common food items than the nest contents of other birds. This, and the typically diverse suite of nest predators in any area, may explain why the rate of predation on artificial nests we placed throughout the urban gradient was not highly correlated with the abundance of crows. We encourage researchers to study how urbanization affects important mechanisms (like nest predators and predation) so environmental policy will benefit from a detailed, scientific understanding of how avian communities are structured. more...

Introduction There is controversy regarding the place of simultaneous pancreas-kidney (SPK) transplantation in end-stage renal disease (ESRD) patients with insulin-dependent diabetes mellitus (IDDM) and detectable c-peptide. We sought to compare outcomes of recipients with and without pretransplantation c-peptide. Methods This retrospective single-center review included consecutive primary SPK transplantations performed between September 2007 and May 2010. Demographic characteristics and outcomes were compared between recipients with and without pretransplantation c-peptide. Results Seven of 25 (28%) consecutive SPK transplant recipients with a diagnosis of IDDM and ESRD had detectable c-peptide prior to transplantation. The mean c-peptide level was 6.3 ± 6.1 ng/mL. For those recipients with and without c-peptide, mean age at diagnosis of IDDM (12.4 ± 7.8 vs 17.1 ± 6.6 years; P = not significant [NS]), duration of IDDM prior to transplantation (30 ± 10 vs 23 ± 9 years; P = NS), and body mass index (25.9 ± 4.5 vs 26.7 ± 4.5 kg/m 2 ; P = NS) were equivalent between the groups. With a median follow-up of 17 months (range, 3–35 months) there was 1 graft loss (due to cardiovascular death) among the 25 patients. At the most recent follow-up, for recipients with and without c-peptide, both the mean serum creatinine (1.3 ± 0.6 vs 1.0 ± 0.2 ng/mL; P = NS) and the mean HbA1c level (5.3 ± 0.4 vs 5.3 ± 0.5; P = NS) were equivalent between the groups. Conclusion For nonobese ESRD patients diagnosed with IDDM at a young age, the presence of detectable c-peptide should not influence the decision to proceed with SPK transplantation. more...

Micro RNAs (miRs) are a family of small noncoding (nc) RNAs, which, similar to other small ncRNA families, such as siRNAs and Piwi RNAs, add a further level of complexity to the regulation of gene expression. miRs represent between 1 and 3% of all ‘genes’, although ncRNAs probably cannot be strictly called ‘genes’. Since their identification in Caenorhabditis elegans in 1993, similar molecules have now been described across the species spectrum, from bacteria (reviewed in Gottesman1) and viruses2 to plants (reviewed in Mallory and Vaucheret3) and animals. Although it has been estimated that roughly 30% of the transcriptome is regulated by miRs, this figure is largely derived from bioinformatics analysis, which can result in both false-positive and false-negative results, and different algorithms may provide only partially overlapping sets of mRNA transcripts that are theoretically regulated by any given miR. The importance of ncRNAs was recognized by the award of the Nobel prize in 2006, and CDD had the honour of hosting the corresponding Nobel lectures.4, 5, 6, 7, 8 In this issue, we focus more specifically on the role of miRs in differentiation and in cell death. more...

Vertebrate responses to human beings have both a learned and genetic component. The learned component is apparently influenced by both the number and outcome of individual human-bird interactions that occur within the animal's lifetime (e.g., Buitron 1983, Knight and Temple 1986). Some bird species have adjusted to high human density and have become "urbanized" (e.g., Cooke 1980). The ability of birds to coexist with humans is largely attributed to the absence of active persecution (e.g., shooting) (Knight et al. 1987). Outside of urban or protected areas, however, active persecution occurs, whether legally (e.g., hunting seasons) or illegally. In the presence of persecution it would be maladaptive to habituate to humans, and birds show avoidance behavior (Knight 1984, Knight et al. 1989). Terrestrial vertebrates may show variation to human intruders by altering flushing response and flight distance (Altmann 1958). Both are important behavioral attributes of organisms in that they affect survival and essential daily routines (e.g., foraging, social behavior). The Black-billed Magpie (Pica pica) is a common speies in Europe and western North America and has made the transition to an urban environment (Birkhead 1991). We examined two hypotheses pertaining to flushing response and flight distance in magpie populations in northeastern Colorado. First, in the absence of active persecution, we predicted that the flushing response and flight distance of magpies would correlate negatively with human density. Second, magpie flushing response and flight distance would be greater in areas with human persecution than in areas without persecution. more...

p53 and its two homologues, p73 and p63, share considerable structural similarities, an ability to interact between themselves and to transactivate the same promoters, including for example p21. Furthermore, p73 can induce cell death via its interaction with c-Abl. In contrast, p63 has been demonstrated to be essential for limb and skin formation. We evaluated the expression of p63 and p73 in differentiating human keratinocytes in vitro. Skin biopsy and primary cultures of normal human epidermal keratinocytes (NHEK) express both p73 and p63. NHEK induced to differentiate in vitro by high calcium exposure show induction of p73 delta and downregulation of all isoforms of p63. This latter gene is predominantly expressed in its transcriptionally inactive form, DeltaNp63. We further evaluated the effect of either p73s or p63 transfected in either NHEK or transformed human keratinocytes (HaCat cells). p73 gamma, delta, and p63 were able to transactivate the promoters of loricrin and involucrin in both NHEK and HaCat cells. These results suggest the involvement of both p73 and p63 genes in keratinocyte terminal differentiation. more...

This study was designed to determine risk factors for intra-abdominal abscess after pancreas transplantation.We performed a single-center retrospective review of all pancreas transplants from 1994 to 1999. Risk factors studied were donor age, body weight, body mass index, peak serum glucose, peak serum amylase, need for pressor agents, cause of death, cold ischemic preservation time, recipient age, type of dialysis, surgical technique, and peak recipient amylase.The 1-year graft survival rate was 90%. Of the 34 patients studied, there were 4 cases of peripancreatic abscess formation (12%). Elevated donor body weight (P0.01), elevated body mass index (P0.05), and the peak recipient serum amylase in the first postoperative week (P0.01) were significant risk factors for the development of intra-abdominal infection.These data suggest that pancreas grafts from obese donors may be more susceptible to ischemia-reperfusion injury resulting in abscess formation. more...

We show here that exposure of cardiac cells to simulated ischemia results in apoptosis and is accompanied by phosphorylation and increased expression and transcriptional activity of STAT-1. Similarly, interferon-gamma, which is known to induce STAT-1 activation, also induced apoptosis in cardiac cells. STAT-1-transfected cells were more susceptible to ischemia-induced cell death than cells transfected with a control plasmid lacking the STAT-1 coding sequence. Furthermore, an antisense STAT-1 vector reduced both ischemia- and overexpressed STAT-1-induced cell death in cardiac cells. Both STAT-1 overexpression and interferon-gamma treatment or exposure to ischemia activated the promoter of the pro-apoptotic caspase-1 gene in cardiomyocytes. Finally, ischemia/reperfusion also induced STAT-1 activation and caspase-1 processing in ventricular myocytes in the intact heart ex vivo. Immunofluorescent staining demonstrated an increase in STAT-1-positive staining in cardiomyocytes in response to ischemia/reperfusion that co-localized with terminal deoxynucleotidyl transferase dVTP nick end-labeling-positive apoptotic cells. These results suggest that STAT-1 plays a critical role in the regulation of ischemia/reperfusion-induced apoptosis in cardiac cells, acting at least in part via a caspase-1 activation-dependent pathway. more...

Urocortin (UCN) is a peptide related to hypothalamic corticotrophin-releasing hormone and binds with high affinity to corticotrophin-releasing hormone receptor-2beta, which is expressed in the heart. In this study, we report that UCN prevented cell death when administered to primary cardiac myocyte cultures both prior to simulated hypoxia/ischemia and at the point of reoxygenation after simulated hypoxia/ischemia. UCN-mediated cell survival was measured by trypan blue exclusion, 3'-OH end labeling of DNA (TUNEL), annexin V, and fluorescence-activated cell sorting. To explore the mechanisms that could be responsible for this effect, we investigated the involvement of MAPK-dependent pathways. UCN caused rapid phosphorylation of ERK1/2-p42/44, and PD98059, which blocks the MEK1-ERK1/2-p42/44 cascade, also inhibited the survival-promoting effect of UCN. Most important, UCN reduced damage in isolated rat hearts ex vivo subjected to regional ischemia/reperfusion, with the protective effect being observed when UCN was given either prior to ischemia or at the time of reperfusion after ischemia. This suggests a novel function of UCN as a cardioprotective agent that could act when given after ischemia, at reperfusion. more...

The primary site of generation on the basilar membrane for the 2f1-f2 distortion product (DP) is generally considered to be near where the higher-frequency stimulus tone peaks. This site has also been shown to be a source of DP otoacoustic emission (DPOAE) in the ear canal, but a second source of emission is known to exist in the region of the DP frequency place. The DPOAE phase versus frequency gradient provides a means of investigating the emission mechanisms. "Wave-fixed" and "place-fixed" mechanisms have been proposed to account for the very different phase gradients found depending on whether the 2f1-f2 DPOAE is evoked by a small or large stimulus-frequency ratio. DPOAE phase versus frequency gradients can be investigated either by sweeping f1,f2 or by sweeping both frequencies maintaining a constant frequency ratio. Each manipulation gives only a partial description of DP behavior. In this study, the place-fixed/ wave-fixed dichotomy is analyzed using extensive 2f1-f2 and 2f2-f1 DP stimulus-frequency sweep data presented on matrices of f1 vs f2 and f2/f1 ratio versus DP frequency. These show how the DPs are related and provide a more complete picture of 2 f1-f2 and 2f2-f1 DPOAE phase and amplitude versus frequency behavior. The phase data contain evidence for a systematic variation in the proportions of wave- and place-fixed emission. The results suggest that 2f1-f2 DPOAEs with a wide stimulus frequency ratio are wave fixed, while all other DPOAEs are place fixed. A transition occurs within the 2 f1-f2 DP data region at a frequency ratio of about f2/f1 = 1.1. The 2f1-f2 DP and 2 f2-f1 DP phase behavior is continuous across the f2/f1 = 1 boundary. As the 2 f2-f1 DP generation region must be strongly influenced by the DP frequency place, the results imply that the place-fixed component of the 2 f1-f2 DP is also linked to its frequency place. A similar pattern was obtained with the 3f1-2f2 and 3f2-2f1 DPs. The results support the following model: For the limited set of stimulus conditions that gives rise to 2 f1-f2 wave-fixed emissions, DP energy is largely generated in the f2 region and is emitted directly. All other DPOAEs are place-fixed emissions, and while nonlinearity within the f2 stimulus envelope remains the generator, the DP is not directly emitted but travels apically until it is re-emitted basally via a separate reflection mechanism in the region of the DP place. more...

Simulated ischaemia causes both necrotic and apoptotic death of primary cultures of neonatal rat cardiac myocytes. Simulated ischaemia is associated with increased expression of urocortin mRNA and with the release of urocortin peptide into the medium. Exogenous urocortin is more potent than corticotropin releasing hormone (CRH) in protecting cardiac myocytes from necrotic and apoptotic death induced by ischaemia, and the cardioprotective effects of ischaemia-preconditioned media are abrogated by antagonists to the CRH family of peptides. Simulated ischaemia increases cardiac myocyte expression of CCAAT enhancer binding (C/EBP) transcription factors, and of the p65 subunit of NFkappaB, and reporter activity of a construct incorporating a fragment of the urocortin promoter containing a C/EBP consensus site is also enhanced by simulated ischaemia. The data suggest that ischaemia, acting partly through increased expression of C/EBP transactivators, increases expression of urocortin mRNA, which is rapidly translated to the mature form. The mature peptide is rapidly released, and exerts autocrine/paracrine protective effects through the cardiac CRH-R2 receptor which preferentially binds urocortin. more...

White Sponge Nevus (WSN) is a rare, autosomal dominant disorder that predominantly affects noncornified stratified squamous epithelia. Clinically, it is characterized by the presence of soft, white, and ‘‘spongy’’ plaques in the oral mucosa. The characteristic histopathologic features are epithelial thickening, parakeratosis, and vacuolization of the suprabasal layer of oral epithelial keratinocytes. Mutations in keratin 4 (K4) and keratin 13 (K13) genes have already been demonstrated to be responsible for WSN; the identification of new keratin mutations in a stratified squamous epithelia closely related to epidermis is of relevance for the understanding of the biochemistry of intermediate filaments, and for genotype phenotype correlations. In this study we investigated a 27-y-old, female Italian patient, affected by white asymptomatic oral plaques. Sequence analysis revealed a 3 bp (ACA) heterozygous insertion localized in the helix initiation motif of the 1A alpha helical domain of K4. We report this new K4 gene mutation and describe an amino acid insertion, in the 1A domain, responsible for a keratin disease. more...

Connective tissue growth factor is a recently described chemoattractant and fibroblast mitogen which, because of sequence homology and weak binding to insulin-like growth factor (IGF)-1, has been proposed as the eighth member of the IGF binding protein (IGFBP) superfamily, named IGFBP-related protein 2 (IGFBP-rP2). Previous studies have implicated IGFBP-rP2 in a number of heterogeneous fibrotic pathologies, including renal fibrosis, dermal scleroderma, and bleomycin-induced pulmonary fibrosis in mice. Because profibrogenic cytokines may be produced by inflammatory cells, we developed a multiplex competitive reverse transcription/polymerase chain reaction to quantify IGFBP-rP2 transcripts in bronchoalveolar lavage cells from healthy subjects and patients with idiopathic pulmonary fibrosis (IPF) and pulmonary sarcoidosis. IGFBP-rP2 messenger RNA expression was enhanced10-fold (P0.003) in patients with IPF;40-fold (P0.006) in stage I/II sarcoidosis patients, and90-fold (P0.005) in stage III/IV sarcoidosis patients by comparison with healthy nonsmoking control subjects. We suggest these increases are predominantly associated with lymphocyte- and neutrophil-driven IGFBP-rP2 production. These findings, together with previous reports implicating other IGFBPs in the pathogenesis of pulmonary fibrosis, suggest that the complex network of IGFBPs within the human lung is an important determinant of the outcome of the fibroproliferative response to injury. more...

We report a case in which en bloc kidneys with bilateral double ureters from a 5-month-old donor were successfully transplanted into a 25-year-old recipient. No stents were used. There were no complications after the transplant. The patient remains well at more than 1.5 years post-transplantation with serum creatinine 1.2 mg/dl. more...

Every so often, research throws up something unexpected. Science being inherently conservative, these reports are usually treated with greater rigour at peer review, and journals are often reluctant to publish contentious papers that do not broadly conform to the established dogma, unless the data are wholly compelling and there is a detailed molecular mechanism. However, withholding potentially heretical material from the wider scientific community deprives it both of intellectual initiative and of the practical opportunity to refute or even substantiate the observations. As might be expected, the more introverted and obsessive referees will recommend major revision with a lot more data, if not outright rejection, whereas the more extrovert and polemical will be more in favour of publication. These papers therefore present Editors with a difficult decision. more...

A simple statistical model of daily precipitation based on the gamma distribution is applied to summer (JJA in Northern Hemisphere, DJF in Southern Hemisphere) data from eight countries: Canada, the United States, Mexico, the former Soviet Union, China, Australia, Norway, and Poland. These constitute more than 40% of the global land mass, and more than 80% of the extratropical land area. It is shown that the shape parameter of this distribution remains relatively stable, while the scale parameter is most variable spatially and temporally. This implies that the changes in mean monthly precipitation totals tend to have the most influence on the heavy precipitation rates in these countries. Observations show that in each country under consideration (except China), mean summer precipitation has increased by at least 5% in the past century. In the USA, Norway, and Australia the frequency of summer precipitation events has also increased, but there is little evidence of such increases in any of the countries considered during the past fiffty years. A scenario is considered, whereby mean summer precipitation increases by 5% with no change in the number of days with precipitation or the shape parameter. When applied in the statistical model, the probability of daily precipitation exceeding 25.4 mm (1 inch) in northern countries (Canada, Norway, Russia, and Poland) or 50.8 mm (2 inches) in mid-latitude countries (the USA, Mexico, China, and Australia) increases by about 20% (nearly four times the increase in mean). The contribution of heavy rains (above these thresholds) to the total 5% increase of precipitation is disproportionally high (up to 50%), while heavy rain usually constitutes a significantly smaller fraction of the precipitation events and totals in extratropical regions (but up to 40% in the tropics, e.g., in southern Mexico). Scenarios with moderate changes in the number of days with precipitation coupled with changes in the scale parameter were also investigated and found to produce smaller increases in heavy rainfall but still support the above conclusions. These scenarios give changes in heavy rainfall which are comparable to those observed and are consistent with the greenhouse-gas-induced increases in heavy precipitation simulated by some climate models for the next century. In regions with adequate data coverage such as the eastern two-thirds of contiguous United States, Norway, eastern Australia, and the European part of the former USSR, the statistical model helps to explain the disproportionate high changes in heavy precipitation which have been observed. more...

Here, we describe a simple methodology which allows the consecutive differentiation of apoptotic and necrotic cells on the same cytospin preparation of fresh and cultured cells. In this methodology, necrotic cells are initially identified on the cytospin preparations using trypan blue followed by the identification of in situ DNA fragmentation using the TUNEL assay. Identification of trypan blue and TUNEL positive cells in the same section permits the simultaneous assessment of necrotic and apoptotic cells. more...

Nitric oxide (NO) modulates the biological activity of proteins by direct interactions with their iron centres. It can also S-nitrosylate cysteines to form S-nitrosothiols. Such reactions affect the activity of membrane-bound, cytosolic and nuclear proteins including the NMDA receptor1, haemoglobin2 and transcription factors such as NF-κB3 and OxyR. NO is potentially toxic, inducing both apoptosis and necrosis. Here we show that NO-mediated S-nitrosylation of the cysteine-containing enzymes that mediate apoptosis (caspases and tissue-transglutaminase, tTG) regulates the balance between apoptosis and necrosis. more...

Aim. To evaluate the outcome of single pediatric kidneys transplanted into adult recipients. Methods. A retrospective single-center review was performed of transplants from donors less than 5 years of age. Outcomes were compared with recipients of grafts from donors 18 to 45 years transplanted during the same time period. Results. Thirty single renal transplants from pediatric donors and 117 transplants from adult donors between 18 and 45 years of age were performed during the study period. The mean age of the pediatric donors was 2.9 ± 0.8 years versus 31.5 ± 8.9 years for adult donors (P more...

Osteoarthritis (OA) is characterized by a loss of joint mobility and pain resulting from progressive destruction and loss of articular cartilage secondary to chondrocyte death and/ or senescence. Certain stimuli including nitric oxide (NO) and the pro-inflammatory cytokine tumor necrosis factor α (TNF-α have been implicated in this chondrocyte death and the subsequent accelerated damage to cartilage. In this study, we demonstrate that a corticotrophin releasing factor (CRF) family peptide, urocortin (Ucn), is produced by a human chondrocyte cell line, C-20/A4, and acts both as an endogenous survival signal and as a cytoprotective agent reducing the induction of apoptosis by NO but not TNF-α when added exogenously. Furthermore, treatment with the NO donor S-nitroso-N-acetyl-𝒟-ℒ-penicillamine upregulates chondrocyte Ucn expression, whereas treatment with TNF-α does not. The chondroprotective effects of Ucn are abolished by both specific ligand depletion (with an anti-Ucn antibody) and by CRF receptor blockade with the pan-CRFR antagonist α-helical CRH(9-41). CRFR expression was confirmed by reverse transcription-PCR with subsequent amplicon sequence analysis and demonstrates that C-20/A4 cells express both CRFR1 and CRFR2, specifically CRFR1α and CRFR2β. Protein expression of these receptors was confirmed by western blotting. The presence of both Ucn and its receptors in these cells, coupled with the induction of Ucn by NO, suggests the existence of an endogenous autocrine/paracrine chondroprotective mechanism against stimuli inducing chondrocyte apoptosis via the intrinsic/mitochondrial pathway. more...

Objective: Myocardial reperfusion following brief periods of ischaemia is associated with prolonged, reversible periods of metabolic dysfunction. As the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is inhibited in vitro by reactive oxygen species, we hypothesized that production of reactive oxygen species during reperfusion would lead to inhibition of GAPDH in post-ischaemic myocardium. Methods: Anaesthetized closed-chest dogs were subjected to 20 min balloon occlusion of the left anterior descending coronary artery. Biopsy samples were taken after 3 and 24 h of reperfusion, to determine the activity of GAPDH and the concentrations of glycolytic intermediates in post-ischaemic and remote, non-ischaemic territories. Results: A significant reduction in GAPDH activity was observed in post-ischaemic relative to remote tissue after 3 h reperfusion (4.8 ± 0.5 vs. 2.9 ± 0.2 μmol/min/mg protein; P < 0.01). Western blotting revealed no reduction in the levels of GAPDH protein. Analysis of enzyme kinetics showed the loss of activity to be associated with decreased V max (5.9 ± 0.5 vs. 3.2 ± 0.2 μmol/min/mg protein; P < 0.01) with no significant change in the K m for glyceraldehyde-3-phosphate (GAP). Incubation of the inhibited enzyme under both mild and strong reducing conditions failed to reactivate the enzyme. The acute reduction in enzyme activity in post-ischaemic tissue was accompanied by regional differences in glycolytic intermediates, notably a twofold accumulation of GAP ( P < 0.05), and a reduction in the glucose metabolic rate (GMR) determined by positron emission tomography and [18F]2-fluorodeoxyglucose. By 24 h reperfusion, no regional differences in GAPDH activity, reaction V max or K m, GAP concentrations or GMR were detectable. Conclusions: These results suggest that inhibition of GAPDH activity may represent an important point at which glycolysis is limited during reperfusion, and further, that the mechanisms of enzyme inhibition do not involve simple oxidation or S -thiolation of critical active site thiol groups. more...

Ischemia and reperfusion lead to the rapid induction of proto-oncogenes in the heart and subsequent induction of genes with cardioprotective functions. The activity of the transcription factors c-Jun and ATF-2 can be stimulated by activation of c-Jun amino-terminal kinase (JNK) in response to a variety of stresses. Here we show that ischemia and reperfusion led to the activation of JNK and also of the distantly-related mitogen activated protein kinase (MAPK). Activation of JNK, but not (MAPK), was abolished by removal of calcium from the perfusate immediately prior to ischemia. In contrast, infusion of the hydrogen peroxide scavenger catalase abolished activation of MAPK in response to ischemia and reperfusion, but activation of JNK was inhibited significantly by catalase only when superoxide dismutase was also present. Hydrogen peroxide infusion activated MAPK but not JNK, supporting a role for hydrogen peroxide produced during reperfusion in MAPK activation. We conclude that while ischemia and reperfusion activate both JNK and MAPK, the mechanisms of activation are different for the 2 kinases. Activation of these kinases is likely to contribute to altered gene expression in response to ischemia and reperfusion. more...

Tumour cells and bacteria share a common property — they frequently acquire resistance to the therapeutics that are administered to treat them. Unfortunately, oncologists and microbiologists often contribute to the problem, as the drugs they use form part of the ongoing selection pressures that can facilitate resistance. In many ways, however, the task of the oncologist is more difficult than that of the microbiologist, as mammalian cells are inherently more complex than their bacterial counterparts. Tumour cells become resistant to therapy by a correspondingly wider range of mechanisms – many of which prevent drug-induced cell death. Three papers and one review1, 2, 3, 4 in this issue of CDD address the problem of tumour resistance to therapy and how we might circumvent this resistance to successfully treat patients with cancer, see Summary. more...

MIRUMIR: an online tool to test microRNAs as biomarkers to predict survival in cancer using multiple clinical data sets

Cardiovascular disease is a leading cause of death worldwide, particularly in Western societies. During an ischaemic insult, ventricular pressure from the heart is diminished as a result of cardiac myocyte death by necrosis and apoptosis. Autophagy is a process whereby cells catabolise intracellular proteins in order to generate ATP in times of stress such as nutrient starvation and hypoxia. Emerging evidence suggests that autophagy plays a positive role in cardiac myocyte survival during periods of cellular stress performing an important damage limitation function. By promoting cell survival, cardiac myocyte loss is reduced thereby minimising the potential of heart failure. In contrast, it has been reported that autophagy can also be a form of cell death. By considering the various animal models of autophagy, we examine the role of the Signal Transducers and Activator of Transcription (STAT) proteins in the autophagic response. Additionally we review the role of the tumour suppressor, p53 and its family member p73 and their potential role in the autophagic response. more...

The effects of a subtherapeutic regimen of rapamycin (RAPA) combined with CsA and donor-specific blood transfusions (DST) were investigated in the ACI to LEW cardiac allograft model. Three protocols were studied. In protocol 1, recipients received a DST (day -1), RAPA intravenously at 0.08 mg/kg/day (day -1 to +5), and CsA intramuscularly at 5 mg/kg/day (day -1 to +1). In protocol 2, the course of CsA was extended to day +5, and in protocol 3 the treatment was further modified to include a second DST on day +3. Control groups received drug treatments alone or combined with DST or third-party blood transfusions. In all protocols, RAPA, CsA, or DSTs alone had little or no effect on graft survival. RAPA-DST or CsA-DST also had no effects beyond the ones induced by the drugs alone. In the RAPA-CsA groups, graft survivals were similar to the ones obtained with CsA alone. In contrast, in each of the protocols, the RAPA-CsA-DST treatment resulted in significant prolongation of graft survival as compared with all controls (P < 0.05). The administration of a second DST (CsA-RAPA-DSTx2) provided no additional benefits over a single DST. Lymph node cells from CsA-RAPA-DST subjects were hyporesponsive in MLR against donor cells and suppressed the proliferation of normal LEW cells in an MLR coculture assay, suggesting the presence of suppressor cells. Furthermore, in the CsA-RAPA-DST group, the anti-donor antibody response was suppressed. These data demonstrate that RAPA, CsA, and DST interact positively by inducing a clear-cut and significant prolongation of allograft survival in the rat model under conditions in which the individual components of the treatment are essentially ineffectual. more...

Analysis of satellite-derived snow cover, radiative balance, and surface air temperature over Northern Hemisphere extratropical land shows that the retreat of the Northern Hemisphere's extent of spring snow cover over the past 20 years parallels a change in the influence of snow cover on the radiative balance and an observed increase of spring temperatures over the same area. These results help explain why the long-term (20th century) increase in surface air temperature over Northern Hemisphere land has been greater in spring than in any other season relative to the interannual variability. more...

Insulin-like growth factor 2 (IGF-2) is the major autocrine growth factor for neuroblastoma. IGF-2 mRNA can just be detected in SK-N-BE(2) cell line; higher levels are present in two clones derived from it [BE(2)-C; BE(2)-M17]. IGF-2 mRNA is increased by retinoic acid (RA) only in the clones. IGF-2 expression/induction is more marked in BE(2)-M17, which shows more RA-resistance (evaluated as growth inhibition, neurite outgrowth and induction of programmed cell death). Under RA exposure, the parental line shows a more pronounced growth inhibition, neurite outgrowth and programmed cell death, as compared to its clones. BE(2)-C cells also express type 1 IGF receptor mRNA, though with a different time course than for expression of IGF-2. The data suggest that IGF-2 expression is correlated with growth, and may counteract the growth retardation, neurite outgrowth and programmed cell death effects of retinoic acid. Therefore the autocrine pattern of IGF-2 production by neuroblastoma cells may promote RA-resistance. more...

A human glioblastoma cell line was found to express in vitro mRNA transcripts specific for insulin-like growth factor-II (IGF-II) and growth-hormone releasing-hormone (GHRH). In the absence of gross morphological changes, retinoic acid reduced the growth rate without major change of IGF-II mRNA expression, while alpha-difluoromethylornithine produced a complete growth arrest and a sharp decrease of IGF-II mRNA expression. Both reagents increased the expression of GHRH mRNA. Also in this glioblastoma cell line, like other neuroectodermal tumours, IGF-II mRNA is expressed independently from GHRH and seems to be parallel to growth rate. more...

The geographic and seasonal pattern of current (1945–1986) zonal mean surface temperature trends has qualitative features in common with ongoing insolation changes caused by perturbations of the earth’s orbit. Temperature and insolation increased in the low latitudes and decreased in the high ones, and showed relative decreases in autumn (SON) as opposed to spring (MAM). The magnitude of the insolation change over the last several decades is too small to have caused the observed temperature trends, which are probably due to internal oscillations of the climate system. However, it is possible that the regional climate trends were amplified when in phase with the ongoing insolation changes and suppressed when out of phase. more...