The toxicity of menthol in short-term bioassays

The toxicity of menthol was studied on 4 different in vitro systems covering organ, cellular and subcellular levels. The 50% inhibitory concentration (IC₅₀) for the cellular and subcellular systems ranged from 0.32 mM to 0.76 mM. At a concentration of 0.5 mM menthol the receptor mediated respiratory stimulation of isolated brown adipocytes was markedly inhibited while the intracellular mitochondrial functions were still unaffected. However, using isolated rat liver mitochondria 0.5 mM menthol was found to cause increase in the ‘state 4’ respiratory rate and osmotic swelling, indicating a leakage of the mitochondrial membrane. We therefore suggest that one effect of menthol is a deterioration of biological membranes. For the determination of the cellular toxicity of foreign compounds isolated brown adipocytes represents a convenient and sensitive model, providing the possibility to localize the primary site of action in terms of mitochondrial or extramitochondrial level.     

Calpain-1 C2L domain peptide protects mouse hippocampus-derived neuronal HT22 cells against glutamate-induced oxytosis

Calpains are Ca²⁺-dependent cysteine proteases; their aberrant activation is associated with several neurodegenerative diseases. The μ-calpain catalytic subunit, calpain-1, is located in the cytoplasm as well as in the mitochondria. Mitochondrial calpain-1 cleaves apoptosis-inducing factor (AIF), leading to apoptotic cell death. We have previously reported that short peptides of calpain-1 C2-like domain conjugated with cell penetrating peptide HIV-Tat (Tat-μCL) selectively inhibit mitochondrial calpain-1 and effectively prevent neurodegenerative diseases of the eye. In this study, we determined whether mitochondrial calpain-1 mediates oxytosis (oxidative glutamate toxicity) in hippocampal HT22 cells using Tat-μCL and newly generated polyhistidine-conjugated μCL peptide and compared their efficacies in preventing oxytosis. TUNEL assay and single strand DNA staining revealed that both μCL peptides inhibited glutamate-induced oxytosis. Additionally, both the peptides suppressed the mitochondrial AIF translocation into the nucleus. All polyhistidine-μCL peptides (containing 4–16 histidine residues) showed higher cell permeability than Tat-μCL. Notably, tetrahistidine (H4)-μCL exerted the highest cytoprotective activity. Thus, H4-μCL may be a potential peptide drug for calpain-1-mediated neurodegenerative diseases such as Alzheimer's disease.     

Parental behaviour of Spanish Imperial Eagles Aquila adalberti: sexual differences in a moderately dimorphic raptor

Capsule The sexes make significantly different contributions in the tasks of nest-building, incubation and food provisioning to chicks.

Aims To determine the division of parental activities during breeding (nest-building, incubation, brooding, food provisioning and feeding).

Methods Between 1991 and 1998 focal observations were made at nests using 20–60× telescopes. During the incubation period, 11 pairs were studied involving a total of 2812 observation hours over 269 days. During the chick-rearing period, seven pairs were studied involving a total of 5499 observation hours spread over 503 days. The birds were sexed and identified individually on the basis of size, position during copulation, vocalization and plumage differences.

Results Significant differences were found between the sexes in parental behaviour. Males scarcely participated in incubation and contributed significantly more than the females only in food provisioning, both during incubation (food transfers to feed the female) and during chick-rearing (to feed the chicks). Females were responsible for most of the incubation, nest-building (during incubation and chick-rearing), brooding, shading and feeding of chicks. Nest attendance by females decreased over time. In both sexes, food provisioning increased with brood size.

Conclusion Intersexual differences are discussed in the context of the reversed sexual size dimorphism and parental investment strategies. We suggest that in Spanish Imperial Eagles reversed sexual size dimorphism is best explained by the prey capture difficulty hypothesis, than by parental role division during reproduction.