Prenatal exposure to testosterone increases ectoparasite susceptibility in the common lizard (Lacerta vivipara)

High levels of testosterone can benefit individual fitness, for example by increasing growth rate or ornament size, which may result in increased reproductive success. However, testosterone induces costs, such as a suppressed immune system, thereby generating trade-offs between growth or mate acquisition, and immunity. In birds and reptiles, females allocate steroids to their eggs, which may be a mechanism whereby females can influence the phenotype of their offspring. To our knowledge, only the benefits of early androgen exposure have been experimentally investigated to date. However, to understand this phenomenon, the costs also need to be evaluated. We manipulated testosterone levels in eggs of the viviparous common lizard and monitored growth, endurance and post-parturient responses to ectoparasites of the offspring. Testosterone-treated individuals had significantly higher growth rates than controls, but suffered a significant decrease in growth rate when exposed to ticks, whereas the corresponding difference for controls was non-significant. There was no difference in observed parasite load or leucocyte count between manipulated and control offspring. Thus, our results suggest that high testosterone levels during embryonic development have detrimental effects on immune function resulting in reduced growth rate, and that this must be taken into consideration when evaluating the potential adaptive value of maternal androgen allocation to eggs.     

Fluorescence imaging of signaling networks

Receptor-triggered signaling processes exhibit complex cross-talk and feedback interactions, with many signaling proteins and second messengers acting locally within the cell. The flow of information in this input-output system can only be understood by tracking where and when local signaling activities are induced. Systematic strategies are therefore needed to measure the localization and translocation of all signaling proteins, and to develop fluorescent biosensors that can track local signaling activities in individual cells. Such a biosensor tool chest can be based on two types of green fluorescent protein constructs that either translocate or undergo fluorescence-resonance-energy transfer when local signaling occurs. Broad strategies to measure quantitative, dynamic parameters in signaling networks, together with perturbation approaches, are needed to develop comprehensive models of signaling networks*.     

Cutting edge: protective cell-mediated immunity to Listeria monocytogenes in the absence of myeloid differentiation factor 88

In addition to their role in triggering innate immune responses, Toll-like receptors are proposed to play a key role in linking the innate and adaptive arms of the immune response. The majority of cellular responses downstream of Toll-like receptors are mediated through the adapter molecule myeloid differentiation factor 88 (MyD88), and mice with a targeted deletion of MyD88 are highly susceptible to bacterial infections, including primary infection with Listeria monocytogenes (LM). In contrast, herein we demonstrate that MyD88-deficient mice have only a modest impairment in their LM-specific CD4 T cell response, and no impairment in their CD8 T cell response following infection with ActA-deficient LM. Furthermore, CD8 T cells from immunized MyD88-deficient mice protected naive recipient mice following adoptive splenocyte transfer, and immunized MyD88-deficient mice were protected from infection with wild-type LM. These results indicate that adaptive immune responses can be generated and provide protective immunity in the absence of MyD88.     

MAHRP-1, a novel Plasmodium falciparum histidine-rich protein,binds ferriprotoporphyrin IX and localizes to the Maurer's clefts

Using a stage-specific cDNA library from Plasmodium falciparum we have identified a gene coding for a novel histidine-rich protein (MAHRP-1). The gene is exclusively transcribed during early erythrocyte stages and codes for a small transmembrane protein. The C-terminal region contains a polymorphic stretch of histidine-rich repeats. Fluorescence microscopy studies using polyclonal mouse sera revealed that MAHRP-1 is located at the Maurer's clefts, which represent parasite-induced structures within the cytosol of infected erythrocytes. Biochemical studies showed that recombinant MAHRP-1 binds the toxic hemoglobin degradation product, ferriprotoporphyrin (FP) with a submicromolar dissociation constant and a stoichiometry determined by the number of DHGH motifs. The bound FP has increased peroxidase-like activity and is 10-fold more susceptible to H2O2-induced degradation compared with unbound FP. These properties of MAHRP-1 suggest it may play a protective role against oxidative stress, and its location at the Maurer's clefts suggests a function in promoting the correct trafficking of exported proteins, such as P. falciparum erythrocyte membrane protein-1.     

Timing and probability of ovulation in relation to sex skin swelling in wild West African chimpanzees, Pan troglodytes verus

Females of many catharrine primates show a periodic and often pronounced swelling of the perineum, the functional significance of which is unclear. Several hypotheses that exist to explain the function of this conspicuous trait are based on assumptions about the temporal relation between the period of maximum swelling and ovulation, and remain largely untested. We examined this relation in free-living chimpanzees of the Ta? National Park, Côte d'Ivoire, and assessed the reliability of perineal swelling as an indicator of ovulation in this species. We used noninvasive urinary progestogen analysis of female reproductive status, together with observational data on swelling characteristics, to determine the variability of timing of ovulation within the maximum swelling phase in 36 cycles from 12 females. The period of maximum swelling was highly variable, lasting from 6 to 18 days. Although ovulation was virtually restricted to the second half of the period of maximum tumescence, its timing varied considerably in relation to both the onset and the end of the maximum tumescence phase. Probability of ovulation, however, was not random, but peaked on day 7 after the onset of the maximum swelling phase, and was almost 60% between days 7 and 9. Thus, in wild chimpanzees perineal swelling indicates the probability of ovulation, but does not provide sufficient information to deduce its exact timing. Given the temporal variability of ovulation relative to the last day of maximum tumescence, field workers should try to include hormonal analysis if information on timing of ovulation is required for interpretation of observational data. Copyright 2003 Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour.      

Molecular dynamics simulations of a pulmonary surfactant protein B peptide in a lipid monolayer

Pulmonary surfactant is a complex mixture of lipids and proteins that lines the air/liquid interface of the alveolar hypophase and confers mechanical stability to the alveoli during the breathing process. The desire to formulate synthetic mixtures for low-cost prophylactic and therapeutic applications has motivated the study of the specific roles and interactions of the different components. All-atom molecular dynamics simulations were carried out on a model system composed of a monolayer of palmitic acid (PA) and a surfactant protein B peptide, SP-B(1-25). A detailed structural characterization as a function of the lipid monolayer specific area revealed that the peptide remains inserted in the monolayer up to values of specific area corresponding to an untilted condensed phase of the the pure palmitic acid monolayer. The system remains stable by altering the conformational order of both the anionic lipid monolayer and the peptide secondary structure. Two elements appear to be key for the constitution of this phase: an electrostatic interaction between the cationic peptide residues with the anionic headgroups, and an exclusion of the aromatic residues on the hydrophobic end of the peptide from the hydrophilic and aqueous regions.     

Regulation of phosducin-like protein by casein kinase 2 and N-terminal splicing

Phosducin-like protein (PhLP) is a member of the phosducin family of G-protein betagamma-regulators and exists in two splice variants. The long isoform PhLP(L) and the short isoform PhLP(S) differ by the presence or absence of an 83-amino acid N terminus. In isolated biochemical assay systems, PhLP(L) is the more potent Gbetagamma-inhibitor, whereas the functional role of PhLP(S) is still unclear. We now report that in intact HEK 293 cells, PhLP(S) inhibited Gbetagamma-induced inositol phosphate generation with approximately 20-fold greater potency than PhLP(L). Radiolabeling of transfected HEK 293 cells with [(32)P] revealed that PhLP(L) is constitutively phosphorylated, whereas PhLP(S) is not. Because PhLP(L) has several consensus sites for the constitutively active kinase casein kinase 2 (CK2) in its N terminus, we tested the phosphorylation of the recombinant proteins by either HEK cell cytosol in the presence or absence of kinase inhibitors or by purified CK2. PhLP(L) was a good CK2 substrate, whereas PhLP(S) and phosducin were not. Progressive truncation and serine/threonine to alanine mutations of the PhLP(L) N terminus identified a serine/threonine cluster (Ser-18/Thr-19/Ser-20) within a small N-terminal region of PhLP(L) (amino acids 5-28) as the site in which PhLP(L) function was modified in HEK 293 cells. In native tissue, PhLP(L) also seems to be regulated by phosphorylation because phosphorylated and non-phosphorylated forms of PhLP(L) were detected in mouse brain and adrenal gland. Moreover, the alternatively spliced isoform PhLP(S) was also found in adrenal tissue. Therefore, the physiological control of G-protein regulation by PhLP seems to involve phosphorylation by CK2 and alternative splicing of the regulator.