Biophysical and biological properties of small linear peptides derived from crotamine, a cationic antimicrobial/antitumoral toxin with cell penetrating and cargo delivery abilities.

Crotamine is a natural polypeptide from snake venom which delivers nucleic acid molecules into cells, besides having pronounced affinity for negatively charged membranes and antifungal activity. We previously demonstrated that crotamine derived short linear peptides were not very effective as antifungal, although the non-structured recombinant crotamine was overridingly more potent compared to the native structured crotamine. Aiming to identify the features necessary for the antifungal activity of crotamine, two linear short peptides, each comprising half of the total positively charged amino acid residues of the full-length crotamine were evaluated here to show that these linear peptides keep the ability to interact with lipid membrane model systems with different phospholipid compositions, even after forming complexes with DNA. Interestingly, the presence of cysteine residues in the structure of these linear peptides highly influenced the antifungal activity, which was not associated to the lipid membrane lytic activity. In addition to the importance of the positive charges, the crucial role of cysteine residues was noticed for these linear analogs of crotamine, although the tridimensional structure and lipid membrane lytic activity observed only for native crotamine was not essential for the antifungal activity. As these peptides still keep the ability to form complexes with DNA molecules with no prejudice to their ability to bind to lipid membranes, they may be potentially advantageous as membrane translocation vector, as they do not show lipid membrane lytic activity and may harbor or not antifungal activity, by keeping or not the semi-essential amino acid cysteine in their sequence.

Hb40-61a: Novel analogues help expanding the knowledge on chemistry, properties and candidacidal action of this bovine α-hemoglobin-derived peptide.

This study expands the knowledge on chemical synthesis and properties of Hb40-61a as well as provides results of the first steps given towards knowing how it kills Candida cells. For the first time, this peptide, its all-D analogue (D-Hb40-61a) and its fluorescently labeled analogue (FAM-Hb40-61a) were successfully assembled on resin at 60°C using conventional heating in all steps. Purified and characterized, these peptides exhibited very low toxicity on human erythrocytes. Hb40-61a and D-Hb40-61a were equally active against Candida strains, ruling out sterically specific interactions on their working mechanism. Cell permeabilization assays confirmed progressive damage of the yeast plasma membrane with increasing concentrations of Hb40-61a. While experiment using the fluorescent probe DiBAC4(5) revealed that this synthetic hemocidin alters the yeast plasma membrane potential, test employing DPH indicated that Hb40-61a might affect its dynamics. Exposure of the yeast cells to FAM-Hb40-61a showed that the peptide accumulates in the cell membrane at the ½ MIC, but stains about 97% of the cells at the MIC. Such effect is salt-dependent and partially energy-dependent. These new findings indicate that the central target of Hb40-61a in Candida cells is the plasma membrane and that this synthetic hemocidin should be considered as a potential candidacidal for topic uses.

Low-dose gamma irradiation of food protein increases its allergenicity in a chronic oral challenge.

Few chronic food protein models have described the relationship between allergenicity and the molecular structure of food protein after physical processing. The effect of γ-radiation on the structure of food protein was measured by fluorescence, circular dichroism and microcalorimetry. BALB/c mice were intraperitoneally sensitized and then given non-irradiated and irradiated Con-A by daily gavage for 28days. The tendency to form insoluble amorphous aggregates and partially unfolded species was observed after irradiation. The administration of non-irradiated and irradiated samples at low-dose significantly increased weight loss as well as plasma levels of eotaxin in animals repeatedly exposed to Con-A. Significant lymphocytic infiltrate filling completely the stroma of microvilli and tubular glands was observed in the small intestinal of the group given Con-A irradiated at a low dose. This phenotype was not observed in animals treated with Con-A irradiated at a high dose.

Opioid blockade improves human recognition memory following physiological arousal.

RATIONALE: States of heightened emotion and arousal, such as those that may occur during crimes or traumatic accidents, can impair human memory. Animal models suggest that such memory alterations may be mediated by opioid neuropeptides. In some experimental paradigms, opioid blockade reverses memory impairments related to arousal. OBJECTIVES: The present study evaluated the hypothesis that, under conditions of heightened arousal, opioid blockade would enhance memory in human subjects. METHODS: Memory for story information was evaluated among subjects randomized to one of four study groups (two orthogonal study conditions): (1) no arousal+no opioid blockade, (2) no arousal+opioid blockade, (3) arousal+no opioid blockade, and (4) arousal+opioid blockade. Both free recall and recognition memory were assessed. Opioid receptor blockade was achieved using a single oral dose of naltrexone. RESULTS: With heightened arousal, subjects receiving naltrexone performed better than those receiving placebo on tests of total and incidental recognition memory. In contrast, with emotionally neutral stimuli, naltrexone subjects performed worse than placebo subjects. CONCLUSIONS: These findings demonstrate that opioid peptides mediate alterations in specific aspects of human memory during heightened emotional states, and help to explain why memories may be selectively deficient under conditions of stress.

Peer support group for adolescents with chronic illness.

This study assessed the effects of a monthly peer support group for adolescents with cancer and other hematological diseases. These adolescents shared activities and experiences with nondisabled high school students. At the group's conclusion, the adolescents reported that the group helped them cope with their illness and improved the quality of their daily lives. Nondisabled students reported that the group favorably affected their attitudes about, and intended behavior toward, peers with chronic illnesses. These results suggest that such groups can provide important benefits for individuals with chronic illnesses as well as for their nondisabled peers.