Impact of ozone on monoterpene emissions and evidence for an isoprene-like antioxidant action of monoterpenes emitted by Quercus ilex leaves

Quercus ilex (L.) leaves emit monoterpenes, particularly alpha-pinene, beta-pinene and sabinene. Apart from the monoterpene pools that are stored in specialized structures and have a clear defensive or attractive role, the function of monoterpenes in Q. ilex leaves is unknown. We tested whether monoterpenes have an antioxidant role, as has previously been found for isoprene in isoprene-emitting leaves. We exposed Q. ilex leaves to either mild and repeated ozone exposure (Experiment I) or to a single acute ozone exposure (Experiment II) at temperatures ranging between 20 and 32 degrees C. Both ozone treatments rapidly stimulated monoterpene synthesis, but had no effect on photosynthesis and caused no visible damage to leaves maintained at 25, 30 or 32 degrees C. Ozone inhibited both photosynthesis and monoterpene synthesis in leaves maintained at 20 degrees C. To characterize the relationship between monoterpenes and ozone-induced damage, we fed detached leaves fosmidomycin, a selective inhibitor of isoprene synthesis. Fosmidomycin caused rapid and complete inhibition of monoterpene emissions in leaves maintained at 30 degrees C, confirming that monoterpenes are synthesized by the same biochemical pathway as isoprene. However, over the experimental period, fosmidomycin did not affect concentrations of compounds that are formed from chloroplastic isoprenoids and that might have conferred antioxidant protection, either directly (carotenoids) or indirectly (chlorophylls, xanthophylls). In leaves whose monoterpene synthesis had been inhibited by fosmidomycin, ozone rapidly and significantly inhibited photosynthesis and increased the production of hydrogen peroxide and malonyldialdehyde. We conclude that monoterpenes produced by Q. ilex leaves share the same biosynthetic pathway and function as isoprene. Furthermore, all volatile isoprenoids may have similar antioxidant properties and may be stimulated by the same stress-inducing conditions.     

Observations on the geographic distribution, ecology and conservation status of several Phaseolus bean species in Costa Rica

Interest in bean genetic resources of Central America has resumed because of disease pressures (e.g., web blight, BGMV) and limitations of current bean varieties. As most of the diversity in landraces has been explored, focus is now on the exploration of wild forms of the primary gene pool and wild species of the secondary gene pool. A germplasm collection was carried out in the field and resulted in the collection of 29 wild populations for six Phaseolus species; it complemented field work done in 1987. Nine more populations were found for P. costaricensis, 10 for wild P. lunatus, one for P. oligospermus, one for P. tuerckheimii, four for wild P. vulgaris and four for P. xanthotrichus. Ninety-three herbarium voucher specimens were collected for 19 populations of the six species (deposited at CR). These results confirm the presence of wild P. vulgaris on both slopes of the central valley of Costa Rica, namely in the life zones bh-MB and bmh-P, and of P. costaricensis in the life zone bmh-MB. These life zones of limited range in Costa Rica have been heavily modified, thus fully justifying the germplasm collection for ex situ conservation. For both species the range of distribution in Costa Rica has been almost completely sampled. The life characteristics of each species that are relevant for their conservation in situ are briefly reviewed. Distribution ranges of each wild bean species are compared with the present extension of national parks, protected areas and fauna/flora sanctuaries, and suggestions for expanding such protected areas are made.     

Physiological responses of Quercus ilex Leaves to Water Stress and Acute Ozone Exposure Under Controlled Conditions

The combined effect of water stress and ozone (O₃) on stomatal O₃ flux, damage to photosynthesis, and detoxification by biogenic volatile organic compounds (BVOC) in Quercus ilex leaves was studied. A 4-weeks O₃ exposure (250 ppb, 4 h per day) caused a reduction of photosynthesis and stomatal conductance, which was fully recovered 1 week after the end of the treatment, in well-watered and water-stressed plants. Measurements of stomatal O₃ flux revealed a low stomatal flux of the pollutant, which became minimal after stomatal closure caused by water stress. An induction of volatile monoterpenes, important compounds in the O₃ scavenging system in Q. ilex, and a burst of lipoxygenase compounds (LOX), which are released as gaseous by-products of membrane peroxidation, was observed after 2–3 weeks of O₃ fumigation. However, these compounds were also released in control leaves that were exposed to ozone only briefly, to determine stomatal O₃ flux. The low stomatal flux that occurred in water stress conditions helped avoiding permanent damage to Q. ilex leaves, although during the O₃ treatment photosynthesis was severely limited by stomatal closure. In well-watered plants, O₃ fumigation caused a noticeable increase of nocturnal stomatal conductance. If confirmed on adult plants under field conditions, this effect can imply larger flux of O₃ at night and possible detrimental effects of O₃ on leaf functions in plants exposed to high nocturnal O₃ levels.     

Diversity and genetic population structure of the Brazilian sharpnose shark Rhizoprionodon lalandii

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Serotonergic Innervation of the Salivary Glands and Central Nervous System of Adult Glossina pallidipes Austen (Diptera: Glossinidae), and the Impact of the Salivary Gland Hypertrophy Virus (GpSGHV) on the Host

Using a serotonin antibody and confocal microscopy, this study reports for the first time direct serotonergic innervation of the muscle sheath covering the secretory region of the salivary glands of adult tsetse fly, Glossina pallidipes Austen. Reports to date, however, note that up until this finding, dipteran species previously studied lack a muscle sheath covering of the secretory region of the salivary glands. Direct innervation of the salivary gland muscle sheath of tsetse would facilitate rapid deployment of saliva into the host, thus delaying a host response. Our results also suggest that the neuronal and abnormal pattern seen in viral infected glands by the Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) is due to a compensatory increased branching of the neurons of the salivary glands, which is associated with the increased size of the salivary glands in viral infected flies. This study shows for the first time serotonin in the cell bodies of the brain and thoracico-abdominal ganglion in adult tsetse, G. pallidipes Austen (Diptera: Glossinidae). A hypothesis is proposed as to whether innervation of the muscle sheath covering of the secretory region of the salivary glands is present in brachyceran compared with nematoceran dipterans; and, a plea is made that more research is needed to develop a blood feeding model, similar to that in the blow flies, for elucidating the various mechanisms involved in production and deployment of saliva.     

A new map of the tiger shark (Galeocerdo cuvier) genetic population structure in the western Atlantic Ocean: Hypothesis of an equatorial convergence centre

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Apoptosis in T lymphocytes from spleen tissue and peripheral blood of L. (L.) chagasi naturally infected dogs

Dogs are the main domestic reservoirs of L. (L.) chagasi. Once in the vertebrate host, the parasite may cause visceral leishmaniasis, which can also be transmitted to humans. Infected symptomatic dogs show disorganization in the white pulp in spleen tissue and a reduction in T lymphocytes in peripheral blood. To investigate whether apoptosis is involved in white pulp disorganization and diminished T cell counts in peripheral blood, apoptotic T cells from the spleen and peripheral blood of dogs naturally infected with L. (L.) chagasi and presenting clinical manifestations were quantified and compared with healthy dogs. Thirteen symptomatic adult dogs infected by L. (L.) chagasi and six healthy dogs from a nonendemic area (controls) were included in the study. Samples from spleen and peripheral blood were used to quantify apoptosis in CD3 lymphocytes by flow cytometry using Anexin V and Multicaspase kits; the results were compared using the Mann Whitney test. The percentage of total T cells was lower in Leishmania infected dogs compared to healthy controls (P<0.05). Apoptosis levels in T cells from PBMC and spleen were higher in infected dogs than in controls (P<0.05). The least squares method test was used to determine the effect between the degree of structural organization of spleen white pulp and the percentage of apoptosis in the spleen. A significant effect on the level of white pulp morphological disorganization and percentage of apoptosis in spleen T cells was observed (F=20.45; P=0.0014). These data suggest that apoptosis is an important for the immunopathogenesis of canine visceral leishmaniasis.     

Flow cytometric detection of alpha-1-acid glycoprotein on feline circulating leucocytes

To assess whether alpha‐1‐acid glycoprotein (AGP) can be detected on the membrane of feline circulating leucocytes. Design The presence of AGP on circulating leucocytes was investigated in both clinically healthy cats and cats with different diseases. A group of feline coronavirus (FCoV)‐positive cats, comprising cats with feline infectious peritonitis (FIP) and cats not affected by FIP but seropositive for FCoV, were included in this study because the serum concentration of AGP increases during FCoV infection. Procedure Flow cytometry (using an anti‐feline AGP antibody), serum protein electrophoresis, routine haematology and measurement of the serum AGP concentration were performed using blood samples from 32 healthy cats (19 FCoV‐seropositive), 13 cats with FIP and 12 with other diseases (6 FCoV‐seropositive). The proportion of cats with AGP‐positive leucocytes in the different groups (e.g. controls vs sick; FIP vs other diseases, etc.) or in cats with different intensities of inflammatory response was compared using a Chi‐square test. Results AGP‐positive leucocytes were found in 23% of cats. Compared with controls, the proportion of patients with positive granulocytes and monocytes was higher among sick cats (especially cats with diseases other than FIP) and cats with high serum AGP concentration, but not in cats with leucocytosis or that were FCoV‐seropositive. Conclusion AGP‐positive leucocytes can be found in feline blood, especially during inflammation. Conversely, no association between AGP‐positive leucocytes and FIP was found. Further studies are needed to elucidate the mechanism responsible for this finding and its diagnostic role in cats with inflammation.