Hypothermic reconditioning by gaseous oxygen persufflation after cold storage of porcine kidneys

BackgroundDelayed graft function still represents a major complication in clinical kidney transplantation. Here we tested the possibility to improve functional outcome of cold stored kidneys a posteriori by hypothermic reconditioning using retrograde oxygen persufflation (ROP) immediately prior to reperfusion.MethodsKidneys from female German Landrace pigs were flushed with Histidine–Tryptophan–Ketoglutarate (HTK) solution and cold-stored for 18 h (control).Some grafts were subsequently subjected to 90 min of retrograde oxygen persufflation (ROP) via the renal vein during cold preservation. Early graft function of all kidneys was assessed thereafter by warm reperfusion in vitro (n = 6, resp.).ResultsRenal function upon reperfusion was significantly enhanced by ROP with an approximately twofold increase in renal clearances of creatinine and urea. ROP also led to higher renal vascular flow rates, enhanced urine output and mitigated histological alterations.ConclusionIt is concluded that initial graft function can be improved by 90 min of hypothermic gaseous oxygenation after arrival of the preserved organ in the transplantation clinic.     

Electrolocation in the presence of jamming signals: behavior

Electrolocation behavior of Apteronotus leptorhynchus was studied by monitoring the animal's ability to maintain orientation to a variety of moving electrolocation targets. The primary goal of this study was to determine the relative effectiveness of various types of electrical 'jamming signals' in disrupting electrolocation performance. 1. Two measures of electrolocation performance were used: The latency between the electrolocation target motion and the fish's following response, and the minimum distance separating the fish from the target during the target movement sequence. Latency increased and minimum fish-target distance decreased as target size was decreased, and when large diameter ceramic targets were used as control stimuli the fish were less able to avoid, and frequently collided with, these 'electrically transparent' objects. 2. Four types of jamming signals were used in attempts to mask the electrosensory input used for electrolocation. Broad-band noise and sinusoidal signals, different in frequency by a few Hz from the animal's personal electric organ discharge (DF stimuli), were used to jam the tuberous electroreceptors. Five Hz and 50 Hz sinusoidal signals were used to jam the low-frequency or ampullary receptor system. Both the noise and the DF stimuli were effective in reducing electrolocation performance, and the threshold intensity for behavior deterioration was about three-fold lower for DF stimuli as compared to the noise. The rate of change of response deterioration as a function of increasing jamming intensity was, however, not different for these two types of stimuli. Neither the 50 Hz nor the 5 Hz jamming signals caused electrolocation deterioration when presented alone. However, 5 Hz jamming, when added to either the noise or DF jamming, did result in significant increments in response deterioration. This suggests that the ampullary receptors can provide supplementary information for electrolocation. 3. Electrolocation performance deterioration was also studied with various difference frequencies between an animal's EOD and the sinusoidal jamming stimulus. Increasing DF results in decreased electrolocation deterioration, but even at the highest DF frequencies used (128 Hz) significant response degradation was observed. 4. The apparent differences in the effectiveness of noise and DF stimulation in reducing electrolocation performance are largely accounted for by the differential effects of the tuberous electroreceptor filter characteristics on these two types of signals.     

Dependence of Intracellular and Exosomal microRNAs on Viral E6/E7 Oncogene Expression in HPV-positive Tumor Cells

Specific types of human papillomaviruses (HPVs) cause cervical cancer. Cervical cancers exhibit aberrant cellular microRNA (miRNA) expression patterns. By genome-wide analyses, we investigate whether the intracellular and exosomal miRNA compositions of HPV-positive cancer cells are dependent on endogenous E6/E7 oncogene expression. Deep sequencing studies combined with qRT-PCR analyses show that E6/E7 silencing significantly affects ten of the 52 most abundant intracellular miRNAs in HPV18-positive HeLa cells, downregulating miR-17-5p, miR-186-5p, miR-378a-3p, miR-378f, miR-629-5p and miR-7-5p, and upregulating miR-143-3p, miR-23a-3p, miR-23b-3p and miR-27b-3p. The effects of E6/E7 silencing on miRNA levels are mainly not dependent on p53 and similarly observed in HPV16-positive SiHa cells. The E6/E7-regulated miRNAs are enriched for species involved in the control of cell proliferation, senescence and apoptosis, suggesting that they contribute to the growth of HPV-positive cancer cells. Consistently, we show that sustained E6/E7 expression is required to maintain the intracellular levels of members of the miR-17~92 cluster, which reduce expression of the anti-proliferative p21 gene in HPV-positive cancer cells. In exosomes secreted by HeLa cells, a distinct seven-miRNA-signature was identified among the most abundant miRNAs, with significant downregulation of let-7d-5p, miR-20a-5p, miR-378a-3p, miR-423-3p, miR-7-5p, miR-92a-3p and upregulation of miR-21-5p, upon E6/E7 silencing. Several of the E6/E7-dependent exosomal miRNAs have also been linked to the control of cell proliferation and apoptosis. This study represents the first global analysis of intracellular and exosomal miRNAs and shows that viral oncogene expression affects the abundance of multiple miRNAs likely contributing to the E6/E7-dependent growth of HPV-positive cancer cells.     

Mixotrophy in orchids: insights from a comparative study of green individuals and nonphotosynthetic individuals of Cephalanthera damasonium

Some green orchids obtain carbon (C) from their mycorrhizal fungi and photosynthesis. This mixotrophy may represent an evolutionary step towards mycoheterotrophic plants fully feeding on fungal C. Here, we report on nonphotosynthetic individuals (albinos) of the green Cephalanthera damasonium that likely represent another evolutionary step. Albino and green individuals from a French population were compared for morphology and fertility, photosynthetic abilities, fungal partners (using microscopy and molecular tools), and nutrient sources (as characterized by 15N and 13C abundances). Albinos did not differ significantly from green individuals in morphology and fertility, but tended to be smaller. They harboured similar fungi, with Thelephoraceae and Cortinariaceae as mycorrhizal partners and few rhizoctonias. Albinos were nonphotosynthetic, fully mycoheterotrophic. Green individuals carried out photosynthesis at compensation point and received almost 50% of their C from fungi. Orchid fungi also colonized surrounding tree roots, likely to be the ultimate C source. Transition to mycoheterotrophy may require several simultaneous adaptations; albinos, by lacking some of them, may have reduced ecological success. This may limit the appearance of cheaters in mycorrhizal networks.     

Cellular Changes in Blood Indicate Severe Respiratory Disease during Influenza Infections in Mice

Influenza A infection is a serious threat to human and animal health. Many of the biological mechanisms of the host-pathogen-interactions are still not well understood and reliable biomarkers indicating the course of the disease are missing. The mouse is a valuable model system enabling us to study the local inflammatory host response and the influence on blood parameters under controlled circumstances. Here, we compared the lung and peripheral changes after PR8 (H1N1) influenza A virus infection in C57BL/6J and DBA/2J mice using virus variants of different pathogenicity resulting in non-lethal and lethal disease. We monitored hematological and immunological parameters revealing that the granulocyte to lymphocyte ratio in the blood represents an early indicator of severe disease progression already two days after influenza A infection in mice. These findings might be relevant to optimize early diagnostic options of severe influenza disease and to monitor successful therapeutic treatment in humans.     

Phenotype and function of engrafted maternal T cells in patients with severe combined immunodeficiency

Engrafted maternal T cells from two patients with severe combined immunodeficiency (SCID) and graft-vs-host disease (GVHD) were characterized for surface phenotype, function, and ecto-5'-nucleotidase (ecto-5'-NT) activity. The majority of engrafted T cells from both patients were T6-, T3+, and Ia+; the ratio of T4+:T8+ cells varied from 0.89 to 3.1 for Patient 1 and was 0.17 for Patient 2. The sum of T4+ + T8+ cells was greater than the number of T3+ cells, and approximately one-third of the patients' T cells were T3-. Two-color immunofluorescent staining showed that one-third of the T cells from Patient 1 had a novel cell surface phenotype (T6-, T3-, T4+, T8+) that was not previously described. T cells from Patient 1 failed to proliferate in response to allogeneic cells or specific antigen and provided little help for PWM-driven Ig synthesis in vitro. However, they did suppress Ig synthesis in vitro and proliferate in response to PHA and Con A; thus they appeared to be more mature than the T cells of Patient 2 and of most previously reported patients with SCID and maternal T cell grafts. Both patients lacked detectable lymphocyte ecto-5'-NT activity, suggesting that either the ecto-5'-NT activity of maternal T cells is lost after engraftment or that a specific subset(s) of ecto-5'-NT-negative maternal T cells predominates in infants with SCID and GVHD. Thus, in vitro T cell function and the proportions of T cells bearing T4 and T8 may vary in SCID patients with maternal T cell grafts. However, the presence of the Ia antigen and the absence of ecto-5'-NT activity may be consistent features of activated maternal T cells responsible for GVHD.