Intracellular Signals Coupled to Muscarinic Acetylcholine Receptor Activation in Cerebral Frontal Cortex from Hypoxic Mice

1. The aim of the present work was to determine hypoxia-induced modifications in the cascade of intracellular events coupled to muscarinic acetylcholine receptor (mAChR) activation in brain. For this purpose, enzymatic activities were measured on normoxically incubated frontal cortical slices from mice exposed to hypobaric hypoxia for 72 hr.2. We found that hypoxia induced alterations in several cerebral enzymatic basal activities: it increased nitric oxide synthase (NOS), but it decreased both membrane protein kinase C (PKC) and phospholipase C activities.3. The mAChR agonist carbachol was found to increase phosphoinositide hydrolysis to greater values in hypoxic tissues than those found in normoxic conditions. Furthermore, a greater translocation of PKC in response to carbachol was observed in hypoxic tissues than in normoxic ones.4. Besides, carbachol induced a drastic reduction of NOS activity in hypoxic brains, at concentrations that stimulated this enzyme activity in normoxic preparations. In the latter, inhibition is obtained only with high concentrations of the cholinergic muscarinicagonist.5. These results pointed to a carbachol-mediated mAChR hyperactivity induced by hypoxic insult.6. The possibility that these effects would account for a compensatory mechanism to diminish NOS hyperactivity, probably protecting for NO neurotoxic action in hypoxic brain, is also discussed.     

New species of bees of Cuba and La Espanola (Hymenoptera: Colletidae,MEgachilidae, Apidae)

Five new species of Antillean bees are described and illustrated: Colletes granpiedrensis n. sp. (Cuba) (Colletidae) is characterized as follows: Head and mesosoma black, legs and metasoma brown. Dense brown hairs on head and mesosoma; white on frons and metasomal terga. Clypeus, frons and mesosoma with large punctures, lesser on vertex and metasoma. Malar space more wide than long. Male and female slightly similar, except in the apical margin of clypeus, supraclipeal area, and color of the pubescence on legs and sterna; Osmia (Diceratosmia) stangei n. sp. (Dominican Republic) (Megachilidae) is characterized as follows: Dark metallic green, metasoma black with metallic green reflections. Pubescence light; body with large, closed punctures. Female with violet reflections in tergum III and mandible tridentate; Coelioxys (Cyrtocoelioxys) alayoi n. sp. (Cuba) (Megachilidae) is characterized as follows: Female black, except basal area of mandibles, tegula, legs, lateral area of tergum I and sterna, reddish brown. Posterior margin of scutellum rounded. Apex of tergum VI with spine curved up. Sternum VI fringed with short, closed setae, and the apex with short spine; Coelioxys (Boreocoelioxys) sannicolarensis n. sp. (Cuba) (Megachilidae) is characterized as follows: Black, except antenna and tegula brown; legs and sterna reddish brown. Clypeal margin straight in profile. Gradular grooves on metasomal terga II and III distinct medially. Fovea on metasomal tergum II of male deep and short, and Triepeolus nisibonensis n. sp. (Dominican Republic) (Apidae) is characterized as follows: Dorsal pubescence (short and dense) on mesosoma and metasoma from white to yellow, according to specimen; ventral pubescence white. Tergal bands of hairs interrupted medially; two spots of light pubescence on tergum II. Male metasomal sternum VII with anterior margin V-shaped and basal constriction narrow.     

Immunochemotherapy in American cutaneous leishmaniasis: immunological aspects before and after treatment

In this study, we evaluated the immune response of patients suffering from cutaneous leishmaniasis treated with two distinct protocols. One group was treated with conventional chemotherapy using pentavalent antimonium salts and the other with immunochemotherapy where a vaccine against cutaneous leishmaniasis was combined with the antimonium salt. Our results show that, although no differences were observed in the necessary time for complete healing of the lesions between the two treatments, peripheral blood mononuclear cells from patients treated by chemotherapy showed smaller lymphoproliferative responses at the end of the treatment than those from patients in the immunochemotherapy group. Furthermore, IFN-gamma production was also different between the two groups. While cells from patients in the chemotherapy group produced more IFN-gamma at the end of treatment, a significant decrease in this cytokine production was associated with healing in the immunochemotherapy group. In addition, IL-10 production was also less intense in this latter group. Finally, an increase in CD8+ -IFN-gamma producing cells was detected in the chemotherapy group. Together these results point to an alternative treatment protocol where healing can be induced with a decreased production of a potentially toxic cytokine.     

MitoTimer

Fluorescent Timer, or DsRed1-E5, is a mutant of the red fluorescent protein, dsRed, in which fluorescence shifts over time from green to red as the protein matures. This molecular clock gives temporal and spatial information on protein turnover. To visualize mitochondrial turnover, we targeted Timer to the mitochondrial matrix with a mitochondrial-targeting sequence (coined “MitoTimer”) and cloned it into a tetracycline-inducible promoter construct to regulate its expression. Here we report characterization of this novel fluorescent reporter for mitochondrial dynamics. Tet-On HEK 293 cells were transfected with pTRE-tight-MitoTimer and production was induced with doxycycline (Dox). Mitochondrial distribution was demonstrated by fluorescence microscopy and verified by subcellular fractionation and western blot analysis. Dox addition for as little as 1 h was sufficient to induce MitoTimer expression within 4 h, with persistence in the mitochondrial fraction for up to 6 d. The color-specific conformation of MitoTimer was stable after fixation with 4% paraformaldehyde. Ratiometric analysis of MitoTimer revealed a time-dependent transition from green to red over 48 h and was amenable to analysis by fluorescence microscopy and flow cytometry of whole cells or isolated mitochondria. A second Dox administration 48 h after the initial induction resulted in a second round of expression of green MitoTimer. The extent of new protein incorporation during a second pulse was increased by administration of a mitochondrial uncoupler or simvastatin, both of which trigger mitophagy and biogenesis. MitoTimer is a novel fluorescent reporter protein that can reveal new insights into mitochondrial dynamics within cells. Coupled with organelle flow cytometry, it offers new opportunities to investigate mitochondrial subpopulations by biochemical or proteomic methods.     

Influence of Cinnamon and Catnip on the Stereotypical Pacing of Oncilla Cats (Leopardus tigrinus) in Captivity

Nonhuman animals in captivity can experience environmental privation that results in their exhibiting abnormal behaviors. Environmental enrichment techniques can help improve their welfare. This study investigated the behavior of 8 zoo-housed oncilla cats (Leopardus tigrinus) in response to 2 odors (catnip and cinnamon) introduced individually into the animals' enclosures for 3 consecutive days. Proportion of scans spent engaging in stereotypical pacing were compared before, during, and after treatments. The addition of cinnamon reduced the proportion of pacing during and after enrichment (Wilcoxon: Z = 3.16, p < .001; Z = 3.16, p < .001, respectively), indicating a prolonged effect of the enrichment on the animals' behavior. Catnip appears to have elicited no significant difference in the stereotypic pacing before, during, or after the enrichment (Friedman: X² = 2.69; p = .260). The results highlight the potential use of cinnamon as a method of environmental enrichment for small captive-housed cats.     

Livestock vehicle accidents in Spain: causes, consequences, and effects on animal welfare

Livestock vehicle accidents are rare but involve significant economic, human, and nonhuman farm animal losses. This study obtained information on the characteristics of accidents, the animals involved, and injuries to humans from newspaper reports about livestock vehicle accidents in Spain from January 2000 to December 2008. Most accidents involved pig transport (57%), followed by bovine (30%), poultry (8%), and sheep (5%). Driver mortality was not high (6%), and most accidents (76%) involved only the livestock vehicle, which often was overturned (64%) on a straight road transect (51%). Multivariate analysis of the data suggests 2 types of accidents, depending on the species transported. In the first cluster, 95.3% of the cases involved pig transport with articulated vehicles (60.5%). In the second cluster, 94.4% of the accidents involved small vehicles used for cattle transport (44.4%). The results of this study indicate that the characteristics of livestock vehicle accidents vary according to species. One of the main causes of accidents appears to be driver fatigue, which may be due to several factors such as intense workdays, poorly designed route plans, or high levels of pressure from companies.     

Uncoupling of osteoblast–osteoclast regulation in a chemical murine model of Gaucher disease

Gaucher disease (GD) is caused by mutations in the GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Type I GD is the mildest form and is characterized by the absence of neuronopathic affection. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood.