Physiographic and climatic events in the Chihuahuan Desert lead to the speciation and distinct demographic patterns of two sister Sceloporus lizards

The biogeographic history of the Chihuahuan Desert is known to be complex, and there is evidence of the effects of physiographic and climatic events in species diversification and demographic population changes in many taxa. Here, using DNA sequence data, we studied the influence of the physiographic and climatic events that occurred in the Chihuahuan Desert during the Pliocene–Pleistocene transition on the speciation and evolutionary history of the sister lizard species Sceloporus cyanostictus and S. gadsdeni. First, based on mtDNA and nDNA sequences, we estimated the divergence times of the sister species. Then, based on mtDNA sequences, we investigated the demographic history of both species within a phylogeographic framework. The divergence time was inferred to be 1.48 Mya, date that corresponds to the existence of a large lake in the Mapimian subprovince, between the current geographic locations of S. cyanostictus and S. gadsdeni. This lake could have acted as a barrier, leading to the speciation of both species. For the demographic history of the two species, we identified two distinct patterns: the population expansion of S. gadsdeni within the Last Glacial Maximum and the potential population decline of S. cyanostictus. Our results can be used as a guide for the study of other aspects that could be critical to developing conservation actions that ensure the survival of not only S. gadsdeni and S. cyanostictus, but also other co‐occurring lizard species.     

Second-harmonic generation from organic molecules incorporated in sol-gel matrices

Orientation of optically nonlinear organic molecules inside sol-gel matrices upon application of an external D.C. electrical field is demonstrated for the first time. The quadratic nonlinear response of silicon oxide or transition metal oxide based gels containing organic molecules has been determined from Electric Field Induced Second Harmonic (EFISH) measurements. Large concentrations of Optically Nonlinear Organic Molecules (ONOM) have been either incorporated inside the macromolecular network or chemically bonded to the oxide backbone of the gels. These results demonstrate the feasibility of permanently poled doped sol-gel matrices. Moreover, EFISH measurements performed on organic molecules appear to be a useful tool for monitoring the changes occurring during sol-gel transformations.     

Fluorescence detection of enzymatically formed hydrogen peroxide in aqueous solution and in reversed micelles

A sensitive enzymatic assay for oxidase reactions both in aqueous solution and in hexadecyltrimethylammoniumbromide (CTAB) reversed micelles has been developed. The assay is based on the fluorescence detection of dichlorofluorescein, which is formed by hydrogen peroxide oxidation of the nonfluorescent precursor dichlorofluorescin. Hydrogen peroxide as product of the reaction catalyzed by glucose oxidase served to select the reaction conditions. The reaction rate is distinctly enhanced in CTAB reversed micelles as compared to the rate in aqueous solution. This effect, combined with the high sensitivity owing to the strong fluorescence of dichlorofluorescein, makes the assay attractive for the detection of low enzyme, substrate, or peroxide concentrations.     

Anatomy, Growth and Survival of a Long-hypocotyl Mutant of Cucumis sativus Deficient in Phytochrome B

Plant dry matter (DM) partitioning, survival rates, stem anatomy,and stem water conductivity were investigated in wild-type (WT)and long-hypocotyl (lh) mutant seedlings of cucumber (Cucumissativus) grown as isolated individuals under natural radiation.The lh mutant is severely deficient in phytochrome B. Wild-typeseedlings accumulated more DM than lh seedlings over a 4-weekgrowth period in the glasshouse. Leaf and root DM were higherin the WT but stem DM was higher in the lh mutant. Stem DM perunit length was larger in WT than in lh mutant seedlings, evenwhen the two genotypes were compared at equal whole plant DM,which was achieved by growing the plants under different irradiance.In WT seedlings, the hypocotyl was shorter but thicker, withlarger average cell diameter than the lh mutant. In hypocotyltransverse sections the area occupied by load-bearing tissues(xylem and phloem fibres) and the number and diameter of xylemvessels were larger in WT than lh seedlings. Survival ratesof the lh mutant were normal in the glasshouse but very lowoutdoors due to hypocotyl fracture. The water conductivity ofhypocotyl sections was higher in WT than lh seedlings, but nosignificant differences in water conductivity were observedwhen the root remained attached to the hypocotyl. These resultssuggest (a) that compared to the WT, tall and slender lh plantsare more susceptible to mechanical stresses created by windimpact, and (b) that if the lh lesion affects the phyB geneonly, phytochrome B plays a role in the elicitation of anatomicaland morphological changes that specifically increase fitnessin open environments.Copyright 1994, 1999 Academic Press Cucumis sativus (cucumber), light phenotypes, phytochrome, photomorphogenesis, shade phenotypes, stem growth     

A Nonintegrative Lentiviral Vector-Based Vaccine Provides Long-Term Sterile Protection against Malaria

Trials testing the RTS,S candidate malaria vaccine and radiation-attenuated sporozoites (RAS) have shown that protective immunity against malaria can be induced and that an effective vaccine is not out of reach. However, longer-term protection and higher protection rates are required to eradicate malaria from the endemic regions. It implies that there is still a need to explore new vaccine strategies. Lentiviral vectors are very potent at inducing strong immunological memory. However their integrative status challenges their safety profile. Eliminating the integration step obviates the risk of insertional oncogenesis. Providing they confer sterile immunity, nonintegrative lentiviral vectors (NILV) hold promise as mass pediatric vaccine by meeting high safety standards. In this study, we have assessed the protective efficacy of NILV against malaria in a robust pre-clinical model. Mice were immunized with NILV encoding Plasmodium yoelii Circumsporozoite Protein (Py CSP) and challenged with sporozoites one month later. In two independent protective efficacy studies, 50% (37.5–62.5) of the animals were fully protected (p = 0.0072 and p = 0.0008 respectively when compared to naive mice). The remaining mice with detectable parasitized red blood cells exhibited a prolonged patency and reduced parasitemia. Moreover, protection was long-lasting with 42.8% sterile protection six months after the last immunization (p = 0.0042). Post-challenge CD8+ T cells to CSP, in contrast to anti-CSP antibodies, were associated with protection (r = −0.6615 and p = 0.0004 between the frequency of IFN-g secreting specific T cells in spleen and parasitemia). However, while NILV and RAS immunizations elicited comparable immunity to CSP, only RAS conferred 100% of sterile protection. Given that a better protection can be anticipated from a multi-antigen vaccine and an optimized vector design, NILV appear as a promising malaria vaccine.     

Degranulation and histamine release from murine mast cells sensitized with dengue virus-immune sera

Mice were immunized with dengue type 2 virus (DEN 2) under a schedule favoring the production of IgE antibody. The antibody obtained could sensitize peritoneal resident mast cells both in vitro and in vivo so that the sensitized cells were degranulated and released histamine on challenge with the DEN 2 antigen. It was also demonstrated that the antibody was cytophilic and heat-labile. The above observations suggest that the present experimental system can be used to detect anti-DEN 2 IgE antibody in mice.     

Volatile profiles of human skin cell cultures in different degrees of senescence

It is known that skin releases volatile organic compounds to the environment, and also that its emission pattern changes with aging of the skin. It could be considered, that these compounds are intermediaries in cell metabolism, since many intermediaries of metabolic pathways have a volatile potential. In this work, a simple and non-destructive method consisting of SPME sampling and GC/MS analysis was developed to identify volatile organic emanations from cell cultures. This technique, applied to skin cells culture, indicates that the cells or cell metabolism produce several skin emissions. Chemometric analysis was performed in order to explore the relationship between a volatile profile and the senescence of cell cultures. Volatile profiles were different for cell cultures in different degrees of senescence, indicating that volatile compound patterns could be used to provide information about the age of skin cells.     

A new tool for determining degrees of mineralization in fossil amphibian skeletons: The example of the Late Palaeozoic branchiosaurid Apateon from the Autun Basin,France

Studying ontogenetic features of fossil tetrapods is of major interest for investigating the adaptive strategies of early tetrapods to their palaeoenvironments. To determine the degree of calcification of skeletal elements, biologists have until now relied on X-ray radiographs of organisms or isolated bones, or on thin sections. An X-ray tomographic scan of Apateon, a Carboniferous – Permian branchiosaurid from the Autun Basin, France, reveals distinct density properties related to different mineralized tissues (calcified cartilage versus bone). The rendering of Apateon as a “test individual” provides a 3D map of the degrees of ossification of the axial and cranial elements. The combination of these anatomical observations with histological information from classical thin sections made in limb bones of several other specimens of the same locality allows the detailed determination of their ontogenetic stage. A comparison with the well-known specimens of the Saar-Nahe Basin, Germany, makes it possible to investigate the influence of different palaeoenvironments on ontogenetic features.