Ascorbic acid prolongs the viability and stability of isolated perfused lungs: A mechanistic study using 31P and hyperpolarized 13C nuclear magnetic resonance

Ex vivo lung perfusion (EVLP) has recently shown promise as a means of more accurately gauging the health of lung grafts and improving graft performance post-transplant. However, reperfusion of ischemic lung promotes the depletion of high-energy compounds and a progressive loss of normal mitochondrial function, and it remains unclear how and to what extent the EVLP approach contributes to this metabolic decline. Although ascorbate has been used to mitigate the effects of ischemia–reperfusion injury, the nature of its effects during EVLP are also not clear. To address these uncertainties, this study monitored the energy status of lungs during EVLP and after the administration of ascorbate using ³¹P and hyperpolarized ¹³C NMR (nuclear magnetic resonance). Our experiments demonstrated that the oxidative phosphorylation capacity and pyruvate dehydrogenase flux of lungs decline during ex vivo perfusion. The addition of ascorbate to the perfusate prolonged lung viability by 80% and increased the hyperpolarized ¹³C bicarbonate signal by a factor of 2.7. The effect of ascorbate is apparently due not to its antioxidant quality but rather to its ability to energize cellular respiration given that it increased the lung’s energy charge significantly, whereas other antioxidants (glutathione and α-lipoic acid) did not alter energy metabolism. During ascorbate administration, inhibition of mitochondrial complex I with rotenone depressed energy charge and shifted the metabolic state of the lung toward glycolysis; reenergizing the electron transport chain with TMPD (N,N,N',N'-tetramethyl-p-phenylenediamine) recovered metabolic activity. This indicates that ascorbate slows the decline of the ex vivo perfused lung’s mitochondrial activity through an independent interaction with the electron transport chain complexes.     

Ex situ transportation of coral larvae for research,conservation, and aquaculture

Due to the lack of appropriate methods to transport high amounts of larvae ex situ over large distances, the availability of coral larvae was so far mainly limited to their place of origin. For a research project at Rotterdam Zoo, The Netherlands, we transported several thousand larvae of three broadcast spawners (Acropora tenuis, A. digitifera, Diploria strigosa) from the Indo Pacific and the Caribbean to Europe. Beside logistics and packing techniques, post-transport survival rates were mainly influenced by larvae density and transport duration. Our results indicate optimum survival rates of >90% at densities of 4 larvae ml⁻¹ when not exceeding a transportation time of 4 days. The ex situ transport of coral larvae over large distances might offer new possibilities for research, conservation, and aquaculture.     

Antimutagenic Acitivity against Trp-P-1 of the Edible Thai Plant,Oroxylum indicum Vent.

A methanolic extract of Oroxylum indicum strongly inhibited the mutagenicity of Trp-P-1 in an Ames test. The major antimutagenic constituent was identified as baicalein with an IC₅₀ value of 2.78±0.15 μM. The potent antimutagenicity of the extract was correlated with the high content (3.95±0.43%, dry weight) of baicalein. Baicalein acted as a desmutagen since it inhibited the N-hydroxylation of Trp-P-2.     

Generation of robust vascular networks from cardiovascular blast populations derived from human induced pluripotent stem cells in vivo and ex vivo organ culture system

Vascular network formation is a key therapeutic event in regenerative medicine because it is essential for mitigating or ameliorating ischemic conditions implicated in various diseases and repair of tissues and organs. In this study, we induced human induced pluripotent stem cells (hiPSCs) to differentiate into heterogeneous cell populations which have abilities to form vascular vessel-like structures by recapitulating the embryonic process of vasculogenesis in vitro. These cell populations, named cardiovascular blast populations (CBPs) in this report, primarily consisted of CD31⁺ and CD90⁺ cells.     

Steps for the Autologous Ex vivo Perfused Porcine Liver-kidney Experiment

The use of ex vivo perfused models can mimic the physiological conditions of the liver for short periods, but to maintain normal homeostasis for an extended perfusion period is challenging. We have added the kidney to our previous ex vivo perfused liver experiment model to reproduce a more accurate physiological state for prolonged experiments without using live animals. Five intact livers and kidneys were retrieved post-mortem from sacrificed pigs on different days and perfused for a minimum of 6 hr. Hourly arterial blood gases were obtained to analyze pH, lactate, glucose and renal parameters. The primary endpoint was to investigate the effect of adding one kidney to the model on the acid base balance, glucose, and electrolyte levels. The result of this liver-kidney experiment was compared to the results of five previous liver only perfusion models. In summary, with the addition of one kidney to the ex vivo liver circuit, hyperglycemia and metabolic acidosis were improved. In addition this model reproduces the physiological and metabolic responses of the liver sufficiently accurately to obviate the need for the use of live animals. The ex vivo liver-kidney perfusion model can be used as an alternative method in organ specific studies. It provides a disconnection from numerous systemic influences and allows specific and accurate adjustments of arterial and venous pressures and flow.     

Conservation status and ex situ cultivation efforts of endemic flora of the Juan Fernández Archipelago

Was realized field studies and ex situ propagation on the vascular flora of Juan Fernández Archipelago during 15 year period. To evaluate the conservation status of a total of 133 species and subspecies of vascular endemic plants I used a IUCN classification founding: 2 species extinct, 1 extinct in it natural habitat, 52 critically endangered, 37 endangered and 9 vulnerable. Thus, 73.8% are contained in a threat category; only 24 taxon can be considered to be of a lesser conservation concern. The largest threat of extinction is a reduction in individuals in local populations resulting in small, isolated populations. This habitat fragmentation and a reduction in endemic flora has also impacted endemic fauna. Besides, during this period was propagated in nurseries a total of 80 of these species and subspecies (60%). It seem clear the necessity to continue to actions conserve this particular ecosystem.     

Genetic variation in the Red-Listed moss Ditrichum cornubicum Paton (Ditrichaceae) and implications for its conservation

Ditrichum cornubicum is a rare and threatened acrocarpous moss found on copper-rich mine waste, characteristic of the EU protected habitat ‘Calaminarian grassland of the Violetalia calaminariae’. The species was once thought to be a British endemic, being known from two former copper/tin mine sites in Cornwall, until the discovery of a population in 2006 at Allihies Mountain Mine, Co. Cork, Ireland, a former copper mine. In light of this discovery, two theories of possible introduction from Britain to Ireland were put forward: (1) an introduction from the 1800s and (2) a more recent introduction (2000s). Only male plants of the species are known, and reproduction and dispersal are therefore solely through asexual propagules and fragmentation. In order to address the conservation questions of the origin of the Irish population and to determine whether diversity exists within this rare species, genetic fingerprinting (amplified fragment length polymorphism) was carried out on the three known global populations. Percent polymorphism was found to be 7.29% and Nei's total gene diversity (H_T) was 0.0356. AMOVA revealed that, of the small amount of variation found, the majority was among the three populations (98%). The presence of five private alleles in the Allihies population suggests a longer period of isolation than would fit either theory of introduction. Cluster analyses reveal that the Allihies population is more genetically distinct than the two British populations are to each other, thus heightening the conservation priority for this population.     

The complete mitogenome of the hydrothermal vent crab Xenograpsus testudinatus (Decapoda, Brachyura) and comparison with brachyuran crabs

In this study, we analyzed the complete mitochondrial (mt) genome of a hydrothermal vent crab Xenograpsus testudinatus (Decapoda: Brachyura) obtained from the hydrothermal vents off Kueishantao Island, Taiwan, which extend from the deep sea Okinawa Trench. The mitogenome of X. testudinatus was 15,796 bp in length and contained the same 37 genes (e.g. 2 rRNAs, 22 tRNAs, and 13 PCGs) found in other metazoan mitogenomes. Analysis of the structural mt gene order in X. testudinatus revealed that the 13 PCGs, excluding a translocation of ND6-Cyt b cluster, were similarly ordered when compared to the pancrustacean ground pattern; however the tRNAs were severely rearranged. Phylogenetic analysis of decapod mitogenomes showed that the molecular taxonomy of the vent crab was in accordance with its morphological systematics. Together, these findings suggest that the vent crab studied here has little mitochondrial genetic variation when compared with morphologically defined conspecifics from other marine habitats.     

Response of endangered plant species to inoculation with arbuscular mycorrhizal fungi and soil bacteria

Three endangered plant species, Plantago atrata and Pulsatilla slavica, which are on the IUCN red list of plants, and Senecio umbrosus, which is extinct in the wild in Poland, were inoculated with soil microorganisms to evaluate their responsiveness to inoculation and to select the most effective microbial consortium for application in conservation projects. Individuals of these taxa were cultivated with (1) native arbuscular mycorrhizal fungi (AMF) isolated from natural habitats of the investigated species, (2) a mixture of AMF strains available in the laboratory, and (3) a combination of AMF lab strains with rhizobacteria. The plants were found to be dependent on AMF for their growth; the mycorrhizal dependency for P. atrata was 91%, S. umbrosus-95%, and P. slavica-65%. The applied inocula did not significantly differ in the stimulation of the growth of P. atrata and S. umbrosus, while in P. slavica, native AMF proved to be the less efficient. We therefore conclude that AMF application can improve the ex situ propagation of these three threatened taxa and may contribute to the success of S. umbrosus reintroduction. A multilevel analysis of chlorophyll a fluorescence transients by the JIP test permitted an in vivo evaluation of plant vitality in terms of biophysical parameters quantifying photosynthetic energy conservation, which was found to be in good agreement with the results concerning physiological parameters. Therefore, the JIP test can be used to evaluate the influence of AMF on endangered plants, with the additional advantage of being applicable in monitoring in a noninvasive way the acclimatization of reintroduced species in nature.