Induction of a Crassulacean acid like metabolism in the C4 succulent plant,Portulaca oleracea L.: physiological and morphological changes are accompanied by specific modifications in phosphoenolpyruvate carboxylase

The induction of a Crassulacean acid like metabolism (CAM) was evidenced after 21–23 days of drought stress in the C₄ succulent plant Portulaca oleracea L. by changes in the CO₂ exchange pattern, in malic acid content and in titratable acidity during the day–night cycle. Light microscopy studies also revealed differences in the leaf structure after the drought treatment. Following the induction of the CAM-like metabolism, the regulatory properties of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31), the enzyme responsible for the diurnal fixation of CO₂ in C₄ plants but nocturnal in CAM plants, were studied. The enzyme from stressed plants showed different kinetic properties with respect to controls, notably its lack of cooperativity, higher sensitivity to L-malate inhibition, higher PEP affinity and lower enzyme content on a protein basis. In both conditions, PEPC's subunit mass was 110 kDa, although changes in the isoelectric point and electrophoretic mobility of the native enzyme were observed. In vivo phosphorylation and native isoelectrofocusing studies indicated variations in the phosphorylation status of the enzyme of samples collected during the night and day, which was clearly different for the control and stressed groups of plants. The results presented suggest that PEPC activity and regulation are modified upon drought stress treatment in a way that allows P. oleracea to perform a CAM-like metabolism. This revised version was published online in August 2006 with corrections to the Cover Date.     

Three-dimensional arrangement of sarcoplasmic reticulum in crayfish as seen by high voltage electron microscope

Summary Sections several m thick of single fibres from the crayfish muscle were examined by means of a high voltage electron microscope to find out the geometry of the sarcoplasmic reticulum (SR). A sufficient contrast of the SR was achieved by lead acetate histochemical method for calcium. Longitudinally oriented files of SR vesicles at the level of A bands and interruptions in otherwise continuous SR net are the most conspicuous features.     

Keratella distribution in North Patagonian lakes (Argentina)

The distribution of Keratella species from 15 different lakes in North Patagonia (Argentina) was analysed. The genus was not present at altitudes above 1000 m. K. tropica was restricted to Patagonian Plateau lakes with a comparatively high conductivity. A morphometric analysis of the widely distributed K. cochlearis was performed. Results showed three groups of K. cochlearis corresponding to Andean lakes, Patagonian Plateau lakes and a Patagonian Reservoir.     

The response of aggregated <Emphasis Type="Italic">Pseudomonas putida</Emphasis> CP1 cells to UV-C and UV-A/B disinfection

UV radiation is a spread method used worldwide for the disinfection of water. However, much of the research on the disinfection of bacterial cells by UV has focused on planktonic cells. Many bacterial cells in nature are present in clumps or aggregates, and these aggregates, which are more resistant to disinfection than their planktonic counterparts, can be problematic in engineered water systems. The current research used Pseudomonas putida (P. putida) CP1, an environmental and non-pathogenic microorganism which autoaggregates when grown under certain conditions, as a model organism to simulate aggregated cells. The study investigated the response of both the planktonic and the aggregated forms of the bacterium to UV-C (λ = 253.7 nm) and UV-A/B (λ > 300 nm) disinfection at laboratory scale in a minimal medium. The planktonic cells of P. putida CP1 were inactivated within 60 s by UV-C and in 60 min by UV-A/B; however, the aggregated cells required 120 min of UV-C treatment and 240 min of UV-A/B radiation to become inactive. The size of the aggregate was reduced following UV treatment. Although all the cells had lost culturability, viability as measured by the LIVE/DEAD^® stain and epifluorescence microscopy was not completely lost and the cells all demonstrated regrowth after overnight incubation in the dark.     

Life cycle assessment of construction and renovation of sewer systems using a detailed inventory tool

Purpose

The objective was to provide comprehensive life cycle inventories for the construction and renovation of sewers. A detailed inventory was provided with multiple options of pipe materials, diameters and site-specific characteristics, and was embedded into the Excel^®-based tool SewerLCA. The tool allows for life cycle evaluation of different sewers. It was applied to determine the most important phases, processes, and related parameters involved in the construction and renovation of sewers from an environmental and economical perspective.

Methods

Comprehensive life cycle inventories (LCIs) for sewers construction and renovation were obtained by first identifying all processes involved after interviewing construction experts and reviewing sewer construction budgets from a Catalan company; and second transforming the processes into masses of materials and energy usage using construction databases. In order to run the life cycle impact assessment (LCIA) the materials and energy typologies from the inventories were matched to their corresponding equivalents into available LCI databases. Afterwards the potential impacts were calculated through the use of LCIA characterization factors from ReCiPe. Life cycle assessment (LCA) was run several times to assess the construction of a 1-km-long sewer with varying pipe materials, life spans for each material, diameters, transport distances, site-specific characteristics, and pipe deposition options.

Results and discussion

The environmental impacts generated by construction and renovation of a 1 km Polyvinylchloride (PVC) pipe with a diameter of 40 cm are mainly associated with pipe laying and backfilling of the trench. The evaluation of several pipe materials and diameters shows that the exclusion of renovation would underestimate the impacts by 38 to 82 % depending on the pipe materials and diameters. Including end-of-life phase for plastic pipe materials increases climate change (up to an extra 71 %) and human toxicity (up to an extra 147 %) impacts (among all diameters). The preferred pipe materials from an environmental point of view are precast concrete and High-Density Polyethylene (HDPE). Site-specific characteristics (specially the presence of rocky soil and asphalt placement) and material life span have a high influence on the overall environmental profile, whereas changes in transport distances have only a minor impact (<4 %).

Conclusions

Environmental impacts during the construction and renovation of sewers are subject to differences in material type, site-specific characteristics and material life span. Renovation of sewers has a large influence on all potential environmental impacts and costs and, hence, should not be omitted in LCA studies. The treatment and disposal processes of plastic pipes at the end of their life has to be accounted in LCA studies.

     

Two-stage culture for producing berberine by cell suspension and shoot cultures of <Emphasis Type="Italic">Berberis buxifolia</Emphasis> Lam

In vitro cultures of Berberis buxifolia were established using thidiazuron (4.5, 23 and 45 mM) or picloram (4 and 40 mM) as plant growth regulators for sustaining growth. For producing berberine, a two-stage culture was performed. In the first step, thidiazuron or picloram were used for biomass production followed by the production stage where benzylaminopurine (4.4 mM) was added as a plant growth regulator. Berberine yields (102 mg g⁻¹ DW) and in vitro shoot cultures (200 mg g⁻¹ DW) were significantly lower than those of whole plants in the field (416 mg g⁻¹ DW). The highest productivity (0.18 mg 1⁻¹ day⁻¹) was attained using picloram (either 4 on 40 mM) in the first stage for producing biomass.     

Factors that affect the re-attachment of Chondracanthus chamissoi (Rhodophyta, Gigartinales) thalli

Vegetative reproduction of Chondracanthus chamissoi by means of fragmentation and re-attachment of thalli is considered an effective strategy for maintaining natural populations of this species. Here, we evaluate the effects of (1) time of drifting thallus, (2) type of substratum, and (3) photon flux density, on the re-attachment capacity of thallus fragments of C. chamissoi. The results show that re-attachment decreases with the time after detachment, and was higher at the lower photon flux densities tested (10 and 40 μmol photons m⁻²s⁻¹), and on calcareous substratum. Secondary attachment discs are formed along the entire surface of the fragment.     

Kairomonal responses of Tomicus destruens (Col., Scolytidae) to host volatiles α-pinene and ethanol

The Mediterranean pine shoot beetle Tomicus destruens is one of the most damaging bark beetles attacking Mediterranean pine forests in southern Europe and north Africa. We studied the attractiveness of the host volatiles α-pinene and ethanol at a range of release rates, alone or in combination, to T. destruens , in order to develop an attractive lure for the management of this beetle. T. destruens was attracted slightly to the host volatile α-pinene, but a strong synergistic effect was found in the attraction towards monoterpene when ethanol was added to the bait. The highest catches of T. destruens were obtained by the optimal blend releasing 300 mg/day of α-pinene and 900 mg/day of ethanol. In contrast to data reported for the related species T. piniperda , Mediterranean pine shoot beetles were clearly attracted to baits releasing ethanol alone (1350 mg/day). trans -Verbenol, which was also added to host volatiles in some tests, did not affect the response. The use of the attractive blend proposed would have a low impact on the natural enemy population of Thanasimus formicarius because of asynchronies in flight periods. Other non-target insects, such as the facultative predator or competitor Oxipleurus nodieri , were also significantly attracted. These results allow the development of an operative lure for the effective monitoring of T. destruens , although improvements by the addition of other host volatiles should be studied.     

Toxoplasma and reaction time: role of toxoplasmosis in the origin, preservation and geographical distribution of Rh blood group polymorphism

The RhD protein which is the RHD gene product and a major component of the Rh blood group system carries the strongest blood group immunogen, the D-antigen. This antigen is absent in a significant minority of the human population (RhD-negatives) due to RHD deletion or alternation. The origin and persistence of this RhD polymorphism is an old evolutionary enigma. Before the advent of modern medicine, the carriers of the rarer allele (e.g. RhD-negative women in the population of RhD-positives or RhD-positive men in the population of RhD-negatives) were at a disadvantage as some of their children (RhD-positive children born to pre-immunized RhD-negative mothers) were at a higher risk of foetal or newborn death or health impairment from haemolytic disease. Therefore, the RhD-polymorphism should be unstable, unless the disadvantage of carriers of the locally less abundant allele is counterbalanced by, for example, higher viability of the heterozygotes. Here we demonstrated for the first time that among Toxoplasma-free subjects the RhD-negative men had faster reaction times than Rh-positive subjects and showed that heterozygous men with both the RhD plus and RhD minus alleles were protected against prolongation of reaction times caused by infection with the common protozoan parasite Toxoplasma gondii. Our results suggest that the balancing selection favouring heterozygotes could explain the origin and stability of the RhD polymorphism. Moreover, an unequal prevalence of toxoplasmosis in different countries could explain pronounced differences in frequencies of RhD-negative phenotype in geographically distinct populations.     

Hyaluronan receptors in the human ocular surface: a descriptive and comparative study of RHAMM and CD44 in tissues, cell lines and freshly collected samples

The purpose of this study was to demonstrate the presence of the receptor for hyaluronan-mediated motility (RHAMM) in human conjunctival epithelium and in two widely used cell lines from human corneal (HCE) and conjunctival (IOBA-NHC) epithelia. We compared the distribution of RHAMM proteins and mRNAs in human ocular surface tissues (corneal, limbal and conjunctival), HCE and IOBA-NHC cell lines, and corneal and conjunctival epithelia primary samples from healthy donors with the previously identified hyaluronan receptor CD44. We also aimed to determine if soluble CD44 (sCD44) was present in human tears, as it could have a role in the interaction of the tear fluid with hyaluronan. Protein expression was evaluated by Western blots and immunofluorescence microscopy. mRNA expression was evaluated by RT-PCR and Q-PCR. sCD44 was analyzed by ELISA in culture supernatants and in human tears. We describe the expression of RHAMM in human healthy conjunctiva and in HCE and IOBA-NHC cells at both protein and mRNA levels, and the presence of sCD44 in human tears. Furthermore, we detected CD44 and sCD44 expression variations in in vitro inflammatory conditions. This study also focused on the necessary caution with which the conclusions extracted from cell lines should be made, and in the great value of using primary samples as often as possible.