Salvia aethiopis L. (Labiatae) roots were collected near Luzón (Guadalajara, Spain) in June 1983. The plant material was dried and a voucher is deposited in the herbarium of the Facultad de Farmacia, Universidad Complutense de Madrid. Aethiopinone (1) and its hemisynthetic derivatives 2 and 3 were obtained as described previously (15, 16) and have been characterized as an o-naphthoquinone diterpene (C20H2402, Mr: 292) and two hemisynthetic products (C39H240, Mr:

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of lesion size is a hard procedure and animals show difficulty to eat, breathe and move because of the lesion. In order to optimize the hamster model for cutaneous leishmaniasis, young adult male and female golden hamsters (Mesocricetus auratus) were injected intradermally at the dorsal skin with 1 to 1.5 x l0 7 promastigotes of Leishmania species and progression of subsequent lesions were evaluated for up to 16 weeks post infection. The golden hamster was selected because it is considered the adequate bio-model to evaluate drugs against Leishmania as they are susceptible to infection by different species. Cutaneous infection of hamsters results in chronic but controlled lesions, and a clinical evolution with signs similar to those observed in humans. Therefore, the establishment of the extent of infection by measuring the size of the lesion according to the area of indurations and ulcers is feasible. This approach has proven its versatility and easy management during inoculation, follow up and characterization of typical lesions (ulcers), application of treatments through different ways and obtaining of clinical samples after different treatments. By using this method the quality of animal life regarding locomotion, search for food and water, play and social activities is also preserved. Video LinkThe video component of this article can be found at http://www.jove.com/video/3533/ Protocol 1. Infection of Hamsters AnimalsInbred female and male golden hamsters (Mesocricetus auratus), 6-8 weeks, weighing 140-160 g are used. They are housed at the animal facility, in temperature-controlled accommodation, fed with standard rodent dried food and provided with water ad libitum. All the procedures involving animals are approved by the institutional Ethical Committee for Experimental Animal Use. Before experimental infection with dermotropic Leishmania parasites animals are sexed, marked and weighted according to standardized procedures. For sexing, animals are inspected for distinctive features such as the visualization of the mammary line and the short ano-genital distance in females, or the visualization of testicles and a greater distance between the anus and foreskin in males. Then, animals are marked by ear piercing or by staining an area of the skin with a swab soaked in picric acid. For ear perforation, after clean with 70% alcohol the ear is pierced using an ear punch for rodents. A region with blood vessels must be avoided. Sedation or anesthesia with a mixture 9:1 of Ketamine (50 mg/kg) and Xilacine (20 mg/kg) intraperitoneally in a volume of 260-300μl 25-G needle is recommended. Finally, animals are weighed by placing them in a trap or box that is conditioned on a precision balance. ParasitesPromastigotes of dermotropic Leishmania species, such as L. amazonensis, are cultured in biphasic Novy-MacNeal-Nicolle (NNN) culture medium at 26°C. Metacyclic (station...

There is a lack of clarity on whether pigs can emotionally respond to musical stimulation and whether that response is related to music structure. Qualitative Behavioral Assessment (QBA) was used to evaluate effects of 16 distinct musical pieces (in terms of harmonic structure) on emotional responses in nursery pigs (n = 30) during four periods: “habituation”, “treatments”, “breaks” and “final”. Data were evaluated using Principal component analysis (PCA). Two principal components (PC) were considered in the analysis: PC1, characterized as a positive emotions index, included the emotional responses content, playful, sociable, and happy, whereas PC2, characterized as a negative emotions index, included fearful, inquisitive, and uneasy with positive loadings, and relaxed and calm with negative loadings. Musical stimulation (treatment) increased (P < 0.01) both emotional indices, compared to other periods and this response was influenced by harmonic characteristics of the music. We concluded that pigs have a wide variety of emotional responses, with different affective states related to the music structure used, providing evidence of its potential use as environmental enrichment for this species.

BackgroundNormal mammary gland contains an extravascular population of B lymphoblasts, precursors of the immunoglobulin plasma cells that play a key role in the passive protection of neonates by secreting immunoglobulins to colostrum and milk. We investigated the presence of chemoattractants in the milk by analysing the chemoattractant activity of various fractions of this secretion. Milk chemoattractants are potentially involved in the recruitment of lymphocytes from the maternal bloodstream in lactating mammary glands.ResultsThe dilution-related lymphoid cell chemoattraction of whey was associated with a < 10 kDa ultrafiltrate. Active fractions were purified by reverse-phase high performance liquid chromatography. Two peptides of 2.7 kDa (DMREANYKNSDKYFHARGNYDAA) and 1 kDa (RPPGLPDKY) were identified as fragments of the SAA protein family, tentatively identified as SAA2. Only the 2.7 kDa synthetic peptide displayed chemotactic activity, at two different optimal concentrations. At the lower concentration (3.7 nM), it attracted B-cell lymphoblasts, whereas at the higher (3.7 μM), it attracted B lymphocytes. Then, the SAA mRNA expression was analysed and we observed more SAA transcripts during lactation than gestation.ConclusionThese data are consistent with the SAA23–45 fragment being involved in preplasma B-cell recruitment to the mammary gland and resultant benefit to the neonate.