Fhit Deficiency-Induced Global Genome Instability Promotes Mutation and Clonal Expansion

Loss of Fhit expression, encoded at chromosome fragile site FRA3B, leads to increased replication stress, genome instability and accumulation of genetic alterations. We have proposed that Fhit is a genome ‘caretaker’ whose loss initiates genome instability in preneoplastic lesions. We have characterized allele copy number alterations and expression changes observed in Fhit-deficient cells in conjunction with alterations in cellular proliferation and exome mutations, using cells from mouse embryo fibroblasts (MEFs), mouse kidney, early and late after establishment in culture, and in response to carcinogen treatment. Fhit ^-/- MEFs escape senescence to become immortal more rapidly than Fhit ^+/+ MEFs; -/- MEFs and kidney cultures show allele losses and gains, while +/+ derived cells show few genomic alterations. Striking alterations in expression of p53, p21, Mcl1 and active caspase 3 occurred in mouse kidney -/- cells during progressive tissue culture passage. To define genomic changes associated with preneoplastic changes in vivo, exome DNAs were sequenced for +/+ and -/- liver tissue after treatment of mice with the carcinogen, 7,12-dimethylbenz[a]anthracene, and for +/+ and -/- kidney cells treated in vitro with this carcinogen. The -/- exome DNAs, in comparison with +/+ DNA, showed small insertions, deletions and point mutations in more genes, some likely related to preneoplastic changes. Thus, Fhit loss provides a ‘mutator’ phenotype, a cellular environment in which mild genome instability permits clonal expansion, through proliferative advantage and escape from apoptosis, in response to pressures to survive.     

Gregariousness affects performance and defensive reactions in swallowtail caterpillars

1. Group living in caterpillars may enhance individual performance due to sharing of costs associated with individual tasks when dealing with biotic or abiotic ecological factors. 2. In the gregarious caterpillar Battus polydamas archidamas (Papilionidae) egg clusters and caterpillar groups vary in size. We hypothesized that individual survival would be higher in larger groups and that group living would enhance individual performance: shorter development time and/or reduced frequency of (presumably costly) defensive reactions in larvae and larger adult size. We also tested whether the group size conferring the highest survival to laboratory-reared caterpillars matched the most frequent egg clutch size in the field. 3. We collected egg clutches in the field and reared caterpillars in groups of 1, 6, 10, and 14 individuals. We quantified larval survival and stage duration as well as adult mass under laboratory conditions, excluding natural enemies. We also recorded the frequency of larval defensive reactions (thrashing and osmeterium display) against a tactile stimulus of first-instar larvae. 4. Group living enhanced caterpillar survival, particularly during the first instars, when caterpillars are 100% gregarious. Groups of intermediate size reduced larval development time but group living did not affect adult mass. Individual caterpillars in groups showed defensive reactions less frequently than solitary individuals, revealing a cost-saving feature of gregariousness for this swallowtail species. The most frequent clutch size in the field (9–10 eggs) did not match the larval group with highest survival (14 individuals), but did match the group with shortest development time.     

Cell death induction in Giardia lamblia: Effect of beta-lapachone and starvation

Giardia lamblia is a protozoan that parasitizes the small intestine of vertebrates. It is a cause of intestinal infection and diarrhea and infects millions of people worldwide. This protozoan presents many characteristics common to eukaryotic cells but it lacks organelles found in most eukaryotes (e.g., peroxisomes, typical Golgi complex and mitochondria). Also it presents mitosomes, a relic organelle that appears to be a mitochondrial remnant. Cell death in Giardia was induced by the drug β-Lapachone and by starvation. Giardia behavior was followed by scanning, transmission and fluorescence microscopy, quantification of cell metabolism using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide), changes in lipid rafts, using DiIC₁₆ and cholera toxin. Cell shrinkage, chromatin condensation, membrane blebbing and vacuolization provided ultrastructural evidence of apoptosis, whereas the myelinic figures in large vacuoles and LC-3 staining suggested an autophagic process. Lipids rafts were altered by drug treatment and co-localized with regions containing membrane blebbing. The treatment with β-Lap induced encystation. A search for sequence similarities in databases and protein alignments was carried out. Although Giardia is an amitochondrial organism, it presented some autophagic-like cell death characteristics and several, but not all, apoptotic characteristics, induced by β-Lapachone and starvation.     

Bioremediation of Soil Contaminated with Diesel Using Inorganic Nitrogen Sources: Incorporating nth-Order Algorithm in the Evaluation of Process Kinetics

In the present study, we investigated the effects of inorganic nitrogen sources—(NPK fertilizer, 15:15:15), (urea fertilizer, 46:0:0), (NH₄)₂SO₄ as well as monitored natural attenuation on the bioremediation of diesel-polluted soil. At the end of the 6-week study, the highest degradation was recorded in soil amended with NPK fertilizer (95 ± 2.77%) while the least total petroleum hydrocarbon removal was observed in monitored natural attenuation (89 ± 2.91%). Nth-order kinetics effectively described three of the treatments out of the four treatment plans. These include urea amendment (r² = 0.9925, average relative error (ARE) = 1.45%, root mean square error (RMSE) = 0.038, k_n = (3.57 ± 0.61) × 10^?2, n = 1.33), NPK fertilizer amendment (r² = 0.9751, ARE = 3.241%, RMSE = 0.086, k_n = (8.04 ± 0.23) × 10^?1, n = 0.74), and monitored natural attenuation (r² = 0.9697, ARE = 2.77%, RMSE = 0.073, k_n = (1.57 ± 0.50) × 10^?2, n = 1.16). The values of n from the nth-order kinetics parameter estimation indicated that all the treatments resulted in diesel degradation that followed a first-order kinetics path. Thus, the outcome of kinetic modeling showed that nth-order can be used as validating tool when many kinetic orders are under consideration. The phytotoxicity assay with Zea mays showed that the treatments plans resulted in germination indices of 17–55%.     

Egg ejection cost can limit defence strategies against brood parasitism

A cost associated with the evolution of antiparasite strategies is the failure to recognize parasitic eggs, leading the host to evict its own eggs. However, there is evidence that birds recognize their own eggs through imprinting. This leads to the question of why birds accept parasitic eggs if such eggs can be identified. Here, we tested whether egg ejection per se can be costly due to increased predation risk to the remaining clutch and whether olfactory or visual cues of egg ejection increase predation. We carried out three field experiments to answer the following questions: (a) Does ejecting an egg increase nest predation risk? (b) Does the presence of olfactory cues, such as the smell of a broken egg, increase nest predation risk? And (c) Does the presence of visual cues, such as an egg shell below the nest, increase nest predation risk? We found evidence that egg ejection increases nest predation and that olfactory cues alone also increase nest predation. The presence of visual cues did not change predation rates. These data indicate that egg ejection is costly for both host and parasitic eggs that may remain in the nest. Our results suggest why host and parasite eggs are commonly found within the same nests, despite the possibility that hosts recognize and could possibly eject the parasite’s egg.