Maternal cannibalistic preferences affect offspring preferences and development in Menochilus sexmaculatus Fabricius (Coleoptera: Coccinellidae): A transgenerational investigation

Cannibalism has been widely reported across taxa. However, the heritability and expression of cannibalistic traits have been least explored. The variation in the expression of cannibalism is likely to exist amongst the population affecting the propensity of cannibalism. Thus, to know whether the mother has any role in the transgenerational transmission of this trait in a ladybird beetle, Menochilus sexmaculatus, we studied the interaction between maternal and offspring prey preferences and its effect on, development duration and body weight of offspring over generations. An insignificant effect of maternal dietary history on offspring prey preference was observed across generations except for the non-cannibalistic adults who significantly preferred aphids over eggs. The long-term detrimental effect of cannibalism was found in cannibals with increased developmental duration and decreased body weight of offspring over generations. In conclusion, the results show that maternal diet did not affect the offspring preferences in M. sexmaculatus but cannibalism had a profound generational effect on the cannibalistic propensity, development and body weight of offspring across generations shows that larval dietary history and nutritional composition of prey contribute to the expression of cannibalistic behaviour across generations.     

Effect of chitosan on physiological, morphological, and ultrastructural characteristics of wood-degrading fungi

An investigation was undertaken to understand the mechanism(s) by which chitosan exerts its antifungal effects against the wood-degrading fungi Sphaeropsis sapinea and Trichoderma harzianum. Exposure to increasing concentrations of chitosan caused an increase in the amount of hydrogen peroxide accumulation in cultures of S. sapinea, which was accompanied by a decrease in superoxide formation. The same effect was not observed in T. harzianum. Potassium ion leakage was an early event for both test fungi, leakage being more pronounced for S. sapinea than T. harzianum for the first 5 min, particularly at higher concentrations of chitosan treatment. Fluorescence microscopy provided evidence that the effect of chitosan on fungal hyphae was mediated through alterations in the plasma membrane properties. Chitosan also severely affected fungal morphology. Increasing concentrations of chitosan induced excessive branching, vacuolation, and a reduction in hyphal diameter. Transmission electron microscopy, which showed more severe ultrastructural changes in S. sapinea hyphae from chitosan treatment as compared to T. harzianum, provided valuable complementary information. The data suggest that the plasma membrane may be the primary target of chitosan action, and that the two fungi differ in the extent to which they are affected.     

Expression of secreted frizzled related protein 1, a Wnt antagonist, in brain, kidney, and skeleton is dispensable for normal embryonic development

Secreted frizzled related protein-1 (sFRP1), an antagonist of Wnt signaling, regulates cell proliferation, differentiation and apoptosis and negatively regulates bone formation. The spatial and temporal pattern of endogenous sFRP1 expression and loss-of-function were examined in the sFRP1-LacZ knock-in mouse (sFRP1-/-) during embryonic development and post-natal growth. beta-gal activity representing sFRP1 expression is robust in brain, skeleton, kidney, eye, spleen, abdomen, heart and somites in early embryos, but sFRP1 gene inactivation in these tissues did not compromise normal embryonic and post-natal development. Kidney histology revealed increased numbers of glomeruli in KO mice, observed after 5 years of breeding. In the skeleton, we show sFRP1 expression is found in relation to the mineralizing front of bone tissue during skeletal development from E15.5 to birth. Trabecular bone volume and bone mineral density in the sFRP1-/- mouse compared to WT was slightly increased during post-natal growth. Calvarial osteoblasts from newborn sFRP1-/- mice exhibited a 20% increase in cell proliferation and differentiation at the early stages of osteoblast maturation. sFRP1 expression was observed in osteoclasts, but this did not affect osteoclast number or activity. These findings have identified functions for sFRP1 in kidney and bone that are not redundant with other sFRPs. In summary, the absence of major organ abnormalities, the enhanced bone formation and a normal life span with no detection of spontaneous tumors suggests that targeting sFRP1 can be used as a therapeutic strategy for increasing bone mass in metabolic bone disorders or promoting fracture healing by modulating Wnt signaling.     

Kinetoplastid membrane protein-11 DNA vaccination induces complete protection against both pentavalent antimonial-sensitive and -resistant strains of Leishmania donovani that correlates with inducible nitric oxide synthase activity and IL-4 generation: evidence for mixed Th1- and Th2-like responses in visceral leishmaniasis

The emergence of an increasing number of Leishmania donovani strains resistant to pentavalent antimonials (SbV), the first line of treatment for visceral leishmaniasis worldwide, accounts for decreasing efficacy of chemotherapeutic interventions. A kinetoplastid membrane protein-11 (KMP-11)-encoding construct protected extremely susceptible golden hamsters from both pentavalent antimony responsive (AG83) and antimony resistant (GE1F8R) virulent L. donovani challenge. All the KMP-11 DNA vaccinated hamsters continued to survive beyond 8 mo postinfection, with the majority showing sterile protection. Vaccinated hamsters showed reversal of T cell anergy with functional IL-2 generation along with vigorous specific anti-KMP-11 CTL-like response. Cytokines known to influence Th1- and Th2-like immune responses hinted toward a complex immune modulation in the presence of a mixed Th1/Th2 response in conferring protection against visceral leishmaniasis. KMP-11 DNA vaccinated hamsters were protected by a surge in IFN-gamma, TNF-alpha, and IL-12 levels along with extreme down-regulation of IL-10. Surprisingly the prototype candidature of IL-4, known as a disease exacerbating cytokine, was found to have a positive correlation to protection. Contrary to some previous reports, inducible NO synthase was actively synthesized by macrophages of the protected hamsters with concomitant high levels of NO production. This is the first report of a vaccine conferring protection to both antimony responsive and resistant Leishmania strains reflecting several aspects of clinical visceral leishmaniasis.     

Identifying metalloproteins through X-ray fluorescence mapping and mass spectrometry

Metals are critical and dynamic components of biochemistry. To understand their roles, we greatly need tools to identify the ligands that bind them within the complexity of natural systems. This work describes the development of methods that not only detect and distinguish metals, but also characterize the proteins that bind them. We describe an approach to expose, identify and quantify metalloproteins in complex mixtures by sequential non-denaturing 2D-gel electrophoresis (2D GE)/X-ray Fluorescence (XRF) and tandem mass spectrometry (MS/MS) in the same spot of a sample. We first apply the development of 2D GE/XRF to Shewanella oneidensis MR-1, a well-studied system, and verify our electrophoretic approach. Then, we identified a novel periplasmic zinc protein in Pseudomonas aeruginosa PAO1 through 2D GE/XRF followed by MS/MS. The identity and function of this protein was verified through a gene mutation experiment.     

Pre-treatment of Pinus radiata substrates by basidiomycetes fungi to enhance enzymatic hydrolysis

Pre-treatment is important step prior to enzymatic hydrolysis of ligno-cellulosic biomass in order to obtain renewable carbon source ca. glucose. Pinus radiata biomass including wood blocks, wood chips and steam exploded wood (SEW) were used to investigate the effect of fungal pre-treatment on glucose yield. Comparison was made using one white-rot fungus (Trametes versicolor) and three brown-rot fungi (Coniophora puteana, Antrodia xantha and Oligoporus placenta). This is the first study where SEW was treated with basidiomycetes and subsequent enzymatic hydrolysis gave 5 g glucose/l which is an order of magnitude greater compared to control biomass (0.5 g glucose/l). This enhanced glucose yield is due to the novel pre-treatment sequence used in this study.     

Plasticity of photosynthetic performance of the Indian tree Butea monosperma TAUB. at three sites with different microclimates

The Fabaceae tree Butea monosperma (TAUB.; syn. Erythrina monosperma (LAM.)) is widely distributed in Central and West-India. We studied it at three sites, i.e. at two locations with contrasting exposure (NE and SW, respectively) in a small mountain range with poor soil on highly drained rocky slopes and at a third location in a plane with deeper soils and better water supply. The two mountain range sites differed in the light climate where the NE-slope obtained more day-integrated irradiance. Chlorophyll fluorescence was measured with a portable fluorometer and leaf samples for stable isotope analyses (δ¹³C, δ¹⁵N, δ¹⁸O) were collected. No differences were seen in carbon and nitrogen contents of leaves at the three sites. N and O isotope signatures of the leaves were similar at the two rocky hill slope sites. More positive values for both signatures were obtained in the leaves in the plane. For all sites saturation of ETR was only achieved well above a PPFD of 1,000 μmol m^?2 s^?1 indicating that the leaves were sun-type leaves. The photosynthetic performance of Butea at the plane was very similar to that at the SW-slope of the mountain range and higher ETRs were obtained at the NE-slope. Ecophysiological flexibility allows Butea to perform well in a variety of habitats and yet gives it particular fitness at specific sites. The best performance was observed in the highly insolated steep rocky hill site (NE-slope) underlining the suitability of the tree for reforestation.     

Cooperation between p27 and p107 during endochondral ossification suggests a genetic pathway controlled by p27 and p130

Pocket proteins and cyclin-dependent kinase (CDK) inhibitors negatively regulate cell proliferation and can promote differentiation. However, which members of these gene families, which cell type they interact in, and what they do to promote differentiation in that cell type during mouse development are largely unknown. To identify the cell types in which p107 and p27 interact, we generated compound mutant mice. These mice were null for p107 and had a deletion in p27 that prevented its binding to cyclin-CDK complexes. Although a fraction of these animals survived into adulthood and looked similar to single p27 mutant mice, a larger number of animals died at birth or within a few weeks thereafter. These animals displayed defects in chondrocyte maturation and endochondral bone formation. Proliferation of chondrocytes was increased, and ectopic ossification was observed. Uncommitted mouse embryo fibroblasts could be induced into the chondrocytic lineage ex vivo, but these cells failed to mature normally. These results demonstrate that p27 carries out overlapping functions with p107 in controlling cell cycle exit during chondrocyte maturation. The phenotypic similarities between p107(-/-) p27(D51/D51) and p107(-/-) p130(-/-) mice and the cells derived from them suggest that p27 and p130 act in an analogous pathway during chondrocyte maturation.