The Effect of Mixing Prunings of Two Tropical Shrub Legumes (Calliandra Houstoniana and Indigofera zollingeriana) with Contrasting Quality on N Release in the Soil and Apparent N Degradation in the Rumen

Lack of synchronization between N released from prunings applied to the soil as green manures and crop uptake as well as optimization of protein digestibility for ruminants, remain major research objectives for the selection of multipurpose tree and shrub legumes (MPT) for mixed smallholder systems in the tropics. Prunings of the high tannin, low quality MPT Calliandra houstoniana CIAT 20400 (Calliandra) and the tannin free, high quality MPT Indigofera zollingeriana (Indigofera) were mixed in the proportions 100:0, 75:25, 50:50, 25:75, and 0:100 (w/w) in order to measure the aerobic rate and extent of N release in a leaching tube experiment, and the anaerobic extent of N degradation in an in vitro gas production experiment. Parameters measured in Calliandra:Indigofera mixtures were compared to theoretical values derived from single species plant material (i.e. 100:0 and 0:100). Aerobic N release and apparent anaerobic N degradation increased with increasing proportion of the high quality legume (Indigofera) in the mixture. While N release in the soil was lower than theoretical values in the mixture 50% Calliandra/50% Indigofera, this was not the case with apparent anaerobic N degradation with the same mixture. Aerobic N immobilization was more pronounced for the mixture 75% Calliandra/25% Indigofera than for 100% Calliandra and negative interaction was observed with apparent anaerobic N degradation in the mixture 75% Calliandra/25% Indigofera. Plant quality parameters that best correlated with aerobic N release and apparent anaerobic N degradation in the rumen were lignin + bound condensed tannins (r=−0.95 and −0.95 respectively, P<0.001). In addition, a positive correlation (r=0.89, P<0.001) was found between aerobic N release in the leaching tube experiment and apparent N degradation in the in vitro anaerobic gas production experiment. Results show that mixing prunings of MPT materials with contrasting quality is an effective way to modify aerobic N release pattern as well as apparent anaerobic N degradation and could possibly be applied to minimize N losses in the rumen and in the soil. In addition, apparent anaerobic N degradation was identified as good predictor of aerobic N release in the soil, which has resource saving implications when screening MTP to be used as green manures.     

Acute toxicity effects of ibuprofen on behaviour and haematological parameters of African catfish Clarias gariepinus (Burchell, 1822)

Indiscriminate discharge of pharmaceutical waste into the aquatic ecosystem may pose serious health challenges to aquatic biota. The effect of acute exposure to ibuprofen was evaluated using changes in behaviour and haematological parameters under static bio-assay method in Clarias gariepinus. Test specimens were exposed to acute concentrations of ibuprofen (0.28, 0.33, 0.38, 0.43 and 0.48 mg l^?1) for 24, 48, 72 and 96 h durations respectively. Behavioural and phenotypic changes were observed in surviving fish. There were significant (p < 0.05) concentration and duration-dependent increases in erythrocyte (RBC), haemoglobin (Hb), pack cell volume (PCV) and leukocytes (WBC) in treated fish compared to the control. Insignificant decreases (p > 0.05) in mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were observed in treated fish compared to the control. Ibuprofen elicited dose and duration- dependent decrease in neutrophil counts with the decreases being significant (p < 0.05) in the higher doses of 0.43 and 0.48 mg l^?1. Ibuprofen did not elicit any significant changes in monocytes, basophils and eosinophils. Changes observed in this study showed that ibuprofen negatively affected the health of the fish and we recommend that discharge of ibuprofen into the aquatic environment should be monitored and controlled.     

Variation in the ribosomal internal transcribed spacers (ITS1 and ITS2) among eight taxa of the Mimulus guttatus species complex

The internal transcribed spacer region (ITS1 and ITS2) of the 18S-25S nuclear ribosomal DNA sequence and the intervening 5.8S region were sequenced from three individuals in each of eight taxa of the Mimulus guttatus species complex. Three discrete variants, or "types," of ITS sequences were found, among which 30%-40% of sites differed, compared with 1%-2% within types. Dot plots indicate that these types were not related by conspicuous rearrangements or inversions. More than one ITS type was often found in the same taxon, and two of three ITS types span species boundaries, indicating their presence prior to speciation. These ITS sequences showed essentially no positional homology with the nearest sequenced relative, tomato. In contrast, the 5.8S region was relatively unvaried, with 8 of 162 sites varied in the sample among all eight taxa. The phylogeny inferred by the most common ITS sequence type, rooted by the two other ITS types, agreed with isozymes in showing the distinctness of M. nudatus, M. laciniatus, and M. tilingii from the other five taxa.      

Hydroxyl Radicals and a Thylakoid-Bound Endopeptidase are Involved in Light- and Oxygen-Induced Proteolysis in Oat Chloroplasts

The hypothesis that light- and oxygen-induced proteolysis inchloroplasts is mediated by active oxygen species was examined.In order to determine whether or not H₂O₂ and/or ^{dot}OH radicalsare involved in these degradative processes we compared thedegradation of proteins in isolated oat chloroplasts exposedto white light at 80 W m^-2 with that in chloroplasts incubatedin darkness in the absence or presence of H₂O₂ or a ^{dot}OH-generatingsystem composed by ascorbic acid, FeCl₃ and H₂O₂ (Asc-Fe-H₂O₂).Light enhanced the rate of degradation of at least 18 polypeptides,while proteolysis was almost negligible in darkness in the abscenceof additives. H₂O₂ had a very small effect. However, Asc-Fe-H₂O₂-treatedchloroplasts in darkness showed a pattern of protein degradationalmost identical to that observed in the light. A thylakoid-boundendopeptidase (EP), the activity of which increased under photooxidativeenvironmental conditions and treatment with an ^{dot}OH-generatingsystem, was partially purified and characterized as a serinetypeprotease. Treatments with inhibitors of serine-type proteaseprevented both light- and Asc- Fe-H₂O₂-induced proteolysis.EP was more active against both soluble and membranous proteinsthat had been pretreated with Asc-Fe-H₂O₂ than against untreatedproteins. It is proposed that a high dose of light irradiationpromotes proteolysis by increasing the formation of ^{dot}OH,which may modify proteins such that they become more susceptibleto EP-catalyzed hydrolysis. ¹Fisiología Vegetal, Dept. de Biología Vegetal,Universidad de Alcalá de Henares, Present address: 28871Alcalá de Henares (Madrid), España.     

Inactivation and Degradation of CuZn-SOD by Active Oxygen Species in Wheat Chloroplasts Exposed to Photooxidative Stress

Changes in CuZn-SOD actvity and content in isolated wheat chloroplastsunder the light, and the involvement of protease(s) and/or activeoxygen species in this process were studied. Both SOD activityand content decayed with exposure time to photooxidative stress.Ascorbate, a H₂O₂ scavenger, prevented photooxidation-associatedinactivation of SOD, while benzoate, a OH scavenger, preventedSOD degradation. Wheat chloroplasts incubated in the dark didnot hydrolyze exogenous or endogenous SOD, either H₂O₂-pretreatedor not. Protease inhibitors did not prevent SOD degradationunder photooxidative treatment, suggesting that plastid protease(s)did not participate in this process. Purified chloroplast CuZn-SODwas exposed to H₂O₂ and or OH-generating systems. had no effect on either SOD activity or stability (estimated bynative PAGE). H₂O₂ up to 700µM inhibited SOD in a dose-dependentmanner and induced charge/mass changes as seen by native PAGE.OH also reduced SOD activity by inducing its fragmentation.High levels of active oxygen, as can be generated under strongstress conditions, could directly inactivate and degrade chloroplasticSOD. ₁ Present address: Departamento de Biología Vegetal,Univer-sidad de Alcalá de Henares, Alcalá de Henares,E-28871 Madrid, Spain     

Changes in the antioxidant metabolism in the embryonic development of the common South American toad Bufo arenarum: differential responses to pesticide in early embryos and autonomous-feeding larvae

Amphibians may be critically challenged by aquatic contaminants during their embryonic development. Many classes of compounds, including organophosphorus pesticides, are able to cause oxidative stress that affects the delicate cellular redox balance regulating tissue modeling. We determined the progression of antioxidant defenses during the embryonic development of the South American common toad, Bufo arenarum. Superoxide dismutase (SOD) and catalase (CAT) activities were high in the unfertilized eggs, and remained constant during the first stages of development. SOD showed a significant increase when the gills were completely active and opercular folds began to form. Reductase (GR) activity was low in the oocytes and increased significantly when gills and mouth were entirely developed and the embryos presented a higher exposure to pro-oxidant conditions suggesting an environmental control. Reduced glutathione (GSH) content was also initially low, and rose continuously pointing out an increasing participation of GSH-related enzymes in the control of oxidative stress. GSH peroxidases and GSH-S-transferases showed relatively high and constant activities, probably related to lipid peroxide control. B. arenarum embryos have plenty of yolk platelets containing lipids, which provide the energy and are actively transferred to the newly synthesized membranes during the early embryonic development. Exposure to the pro-oxidant pesticide malathion during 48 h did not significantly affect the activity of antioxidant enzymes in early embryos, but decreased the activities of CAT, GR, and the pool of GSH in larvae. Previous work indicated that lipid peroxide levels were kept low in malathion-exposed larvae, thus we conclude that oxidative stress is overcome by the antioxidant defenses. The increase in the antioxidant metabolism observed in the posthatching phase of development of B. arenarum embryo, thus constitutes a defense against natural and human-generated pro-oxidants present in the aquatic environment.     

Physiological characterization of four model Lotus diploid genotypes: L. japonicus (MG20 and Gifu), L. filicaulis, and L. burttii under salt stress

The genus Lotus comprises a heterogeneous group of annual and perennial species. Lotus japonicus (with MG20 and Gifu ecotypes) has been adopted as one of the model legumes in genetic and genomic studies. Other Lotus species, such us Lotus burttii and Lotus filicaulis, have also been used in genetic and genomic studies because of their capacity to produce fertile progenies in crosses with L. japonicus. In the present work, physiological responses to salt stress in four Lotus genotypes were evaluated on the basis of growth and associated parameters, such as photosynthesis, ions, relative water content, oxidative damage and antioxidant system responses, using two NaCl levels applied by acclimation for up to 28 and 60 d. Growth responses varied with plant developmental stage in the four Lotus genotypes. L. japonicus MG20 was found to be a salt-tolerant genotype, mainly when exposed to salt stress at the young plant stage. The capacity of Lj MG20 to sustain growth under salt stress was correlated with enhancement of Superoxide dismutase and Glutathione reductase activities, as well as with increases in total and reduced glutathione content and lower Na⁺ accumulation in leaves. These results suggest that enhancement of antioxidant responses in Lj MG20 contributed to improve salt stress tolerance at early stages. On the other hand, after long-term high NaCl stress treatment, L. filicaulis exhibited lower biomass reduction, lower oxidative damage and Na⁺ accumulation in leaves than the control treatment; hence, this genotype was considered salt-tolerant. These apparently ambiguous results remark that salt tolerance, as a development-related process, was differentially expressed among the Lotus genotypes and depended on stress duration and plant phenological stage.     

Isocitrate dehydrogenase 1 R132C mutation occurs exclusively in microsatellite stable colorectal cancers with the CpG island methylator phenotype

The CpG Island Methylator Phenotype (CIMP) is fundamental to an important subset of colorectal cancer; however, its cause is unknown. CIMP is associated with microsatellite instability but is also found in BRAF mutant microsatellite stable cancers that are associated with poor prognosis. The isocitrate dehydrogenase 1 (IDH1) gene causes CIMP in glioma due to an activating mutation that produces the 2-hydroxyglutarate oncometabolite. We therefore examined IDH1 alteration as a potential cause of CIMP in colorectal cancer. The IDH1 mutational hotspot was screened in 86 CIMP-positive and 80 CIMP-negative cancers. The entire coding sequence was examined in 81 CIMP-positive colorectal cancers. Forty-seven cancers varying by CIMP-status and IDH1 mutation status were examined using Illumina 450K DNA methylation microarrays. The R132C IDH1 mutation was detected in 4/166 cancers. All IDH1 mutations were in CIMP cancers that were BRAF mutant and microsatellite stable (4/45, 8.9%). Unsupervised hierarchical cluster analysis identified an IDH1 mutation-like methylation signature in approximately half of the CIMP-positive cancers. IDH1 mutation appears to cause CIMP in a small proportion of BRAF mutant, microsatellite stable colorectal cancers. This study provides a precedent that a single gene mutation may cause CIMP in colorectal cancer, and that this will be associated with a specific epigenetic signature and clinicopathological features.     

Isonymic relationships in ethno-social categories (Argentinian colonial period) including illegitimate reproduction

Surnames provide a useful method to study the structure of human populations for which biological data are not available. The isonymic method has had multiple applications, but difficulties emerge when dealing with groups where extramarital reproduction is common and the sample size is small, and even more so when only paternal surnames are taken into account.Therefore, it could be of interest to retain female surnames, including those of unmarried mothers. This study was carried out using all birth records froman Argentinian population in the colonial period, which was characterized by the presence of different ethno-social groups (Spanish, Indian and 'Mestizo'or mixed Spanish-Indian) and various reproductive patterns regarding legitimacy. Coefficient of relationship by isonymy (Ri) kinship matrices between geographical populations were obtained, and the results derived from sets of surnames (paternal, maternal of legitimate and illegitimate children,and all surnames in the registers) compared. The results show similar surname distribution regardless of the set of surnames and group considered.Kinship Ri matrices using paternal surnames, maternal surnames of legitimate children, maternal surnames of illegitimate children, and the set of whole surnames showed the same relationships among populations, indicating a similar pattern for Spanish, Indian and Mixed ethno-social groups. Mantel test correlation between all pairs of matrices was significant in all different ethno-social groups. The results suggest that in populations with high illegitimacy, such as that studied here, it is possible to include maternal surnames, even corresponding to single mothers, in order to consider total reproduction and therefore maximize sample size.     

Variation in heat shock proteins within tropical and desert species of poeciliid fishes

The 70-kilodalton heat shock protein (hsp70) family of molecular chaperones, which contains both stress-inducible and normally abundant constitutive members, is highly conserved across distantly related taxa. Analysis of this protein family in individuals from an outbred population of tropical topminnows, Poeciliopsis gracilis, showed that while constitutive hsp70 family members showed no variation in protein isoforms, inducibly synthesized hsp70 was polymorphic. Several species of Poeciliopsis adapted to desert environments exhibited lower levels of inducible hsp70 polymorphism than the tropical species, but constitutive forms were identical to those in P. gracilis, as they were in the confamilial species Gambusia affinis. These differences suggest that inducible and constitutive members of this family are under different evolutionary constraints and may indicate differences in their function within the cell. Also, northern desert species of Poeciliopsis synthesize a subset of the inducible hsp70 isoforms seen in tropical species. This distribution supports the theory that ancestral tropical fish migrated northward and colonized desert streams; the subsequent decrease in variation of inducible hsp70 may have been due to genetic drift or a consequence of adaptation to the desert environment. Higher levels of variability were found when the 30- kilodalton heat shock protein (hsp30) family was analyzed within different strains of two desert species of Poeciliopsis and also in wild-caught individuals of Gambusia affinis. In both cases the distribution of hsp30 isoform diversity was similar to that seen previously with allozyme polymorphisms.