Histoire plio-pléistocène des écosystèmes végétaux de Méditerranée sud-occidentale : apport de l’analyse pollinique de deux sondages en mer d’Alboran

The pollen analysis of sediments of two boreholes located in the northeastern Morocco (Nador 1) and in the southeastern Spain (Andalucia G1) has allowed shedding a new light on the vegetation and climate of the North African littoral plains, the Rif Massif and the Betic Range during the Pliocene. The vegetation around the Alboran Sea was open and xeric during the Zanclean, dominated by herbs including subdesertic elements as Calligonum, Lygeum, Nitraria and Neurada. This type of vegetation indicates a dry and hot climate. The southwestern Mediterranean steppes have therefore a climatic character; they existed before the presence and the heavy pressure of Man on the environment. From the Piacenzian, the development, at Andalucia as well as at Nador, of Artemisia and the appearance of some altitude trees such as Cedrus and Cathaya indicate a vegetation change linked to a climatic change. Modifications in the vegetation observed during the same period in northwestern Mediterranean seem to indicate that the vegetation changes observed at Andalucia and Nador are controlled by the appearance of the first arctic glacial-interglacial cycles. The cedar tree appears at Nador only at the end of the Piacenzian, at time of the first cooling, while it has been found punctually in Messinian and early Pliocene sediments of Habibas in Algeria and in Messinian sediments of the Bou Regreg section at Salé in Morocco. The regular presence of Cedar at Bou Regreg during the Messinian, allows supposing that it was present in the Middle Atlas Mountains and that its development was favoured by colder conditions.     

Susceptibility of<Emphasis Type="Italic">Escherichia coli</Emphasis> to C<Subscript>2</Subscript>-C<Subscript>18</Subscript> fatty acids

The antimicrobial activity of C2-C18 fatty acids was determined in vitro in cultures of two strains of Escherichia coli grown on glucose. Antimicrobial activity was expressed as IC50 (a concentration at which only 50% of the initial glucose in the cultures was utilized). Utilization of glucose was inhibited by caprylic acid (IC50 0.30-0.85 g/L) and capric acid (IC50 1.25-2.03 g/L). Neither short-chain fatty acids (C2-C6) nor fatty acids with longer chain (C12-C18) influenced substrate utilization. Caproic acid, however, decreased cell yield in cultures of E. coli in a dose-dependent manner. No inhibition of glucose utilization was produced with unsaturated fatty acids, oleic and linoleic. Calcium ions added in excess reversed the antimicrobial effect of capric acid, but not that of caprylic acid. Antimicrobial activity of caprylic and capric acid decreased when the bacteria were grown in the presence of straw particles, or repeatedly subcultured in a medium containing these compounds at low concentrations. Counts of viable bacteria determined by plating decreased after incubation with caprylic and capric acid (30 min; 1 g/L) at pH 5.2 from > 10(9) to approximately 10(2)/mL. A reduction of a mere 0.94-1.96 log10 CFU was observed at pH 6.5-6.6. It can be concluded that caprylic acid, and to a lesser extent also capric acid, has a significant antimicrobial activity toward E. coli. Effects of other fatty acids were not significant or absent.     

Coal-smoke pollution modifies physio-chemical characteristics of tissues during the ontogeny of <Emphasis Type="Italic">Peristrophe bicalyculata</Emphasis>

Coal-smoke emissions of a thermal power plant affected the physio-chemical status of Peristrophe bicalyculata (Reth) Nees, as observed at the pre-flowering, flowering and post-flowering stages of plant growth. The nitrate level was raised while nitrate reductase activity, and the soluble protein content of leaf declined heavily at the polluted site during different stages of plant growth, compared to the control. The rate of photosynthesis also decreased under the pollution stress. Sugar level in root, stem and leaves increased with growing age of the plant but was always lower at the polluted site than at the reference site. In roots, the difference was marginal till flowering stage and quite conspicuous afterwards; stems showed a reverse pattern of variation. Sulphur content was higher at the polluted site in all the organs and at each stage of the plant life. The Zn and Fe concentrations were reduced in all plant parts under the pollution stress. Copper content in root was consistently low at the polluted site. In the stem and leaves, however, it was almost equal on both the sites at the pre-flowering stage but showed a wide difference during the later part of plant ontogeny.     

Regeneration of Carnegiea gigantea (Cactaceae) since 1850 in three populations in the northern Sonoran Desert

Saguaro cacti (Carnegiea gigantea) are long-lived and exhibit great variability in growth that makes age estimation problematic. A few single-site studies have focused on those locales where long-term data (e.g. 85 years) are available. Using a newly developed technique, 733 saguaros were sampled in three locales (Silverbell, Harcuvar, Kofa) across Arizona and their age structure reconstructed for the last 150 years based on a mathematical model of the heights of individuals. This is the first study to compare regeneration at multiple locations across the species' range. Regression analysis for each site (years and frequency of individuals established during that year) was run and residuals extracted to determine peaks and troughs in regeneration over time. Correlation was run on the residuals between sites, and chi-square analysis was employed to compare frequency of good and bad regeneration years between Kofa and Silverbell. Peaks and troughs represent regeneration as well as survivorship and mortality. Several large cohorts established at Kofa and Harcuvar since 1850, while at Silverbell well over 80% of sampled saguaros established after the late 1930s. This more recent recruitment at Silverbell may be related to the major freezing event of 1937 whose impact was likely greater at the cooler Silverbell site. Despite the widely different population structures at Silverbell and Kofa, recruitment trends in both populations were statistically linked as both locales often benefited from the same favorable periods for regeneration (P < 0.001). The Harcuvar population shares some common peaks and troughs in regeneration over time with Kofa and Silverbell, but its relationship to them is not statistically significant. Some trends overlap in some locales, such as the favorable regeneration period in the late 1800s and early 1900s, particularly at Silverbell and Kofa as well as at other known sites. However, each population has its own signature. Silverbell is a youthful population likely shaped by severe freezing events, while Kofa has many individuals representing regeneration at several different periods. Regeneration, mortality and subsequent population structure is shaped by both regional-scale influences as well as more localized conditions over the long and short terms.     

Stress-protective activity of the CH<Subscript>3</Subscript>CO-Lys-Lys-Arg-Arg-NH<Subscript>2</Subscript> synthetic peptide (protectin)

The CH₃CO-Lys-Lys-Arg-Arg-NH₂ peptide (the author has named it protectin) was synthesized, and its activity was studied during different stress actions. Protectin was found to normalize the content of corticosterone and adrenalin in adrenal glands and blood after its intranasal administration to rats one day before a cold or heat shock, or hypobaric hypoxia at doses of 1–10 μg/animal and after its intravenous administration just after acute hemorrhage at doses of 0.5–2 μg/animal. The intranasal administration of protectin at doses of 1–10 μg/rat one day before the heat or cold shock was also shown to prevent a change in the content of free histamine and the activity of diamine oxidase in myocardium, which was induced by the dramatic change in the activity of the enzyme after the temperature actions.     

Gene cloning,purification, and characterization of recombinant DNA ligases of the thermophilic archaea <Emphasis Type="Italic">Pyrococcus abyssi</Emphasis> and <Emphasis Type="Italic">Methanobacterium thermoautotrophicum</Emphasis>

DNA ligase genes of the thermophilic archaeae Pyrococcus abyssi (Pab DNA ligase) and Methanobacterium thermoautotrophicum (Mth DNA ligase) were cloned in Escherichia coli. The resulting recombinant enzymes were tested for activity in a ligation mixture with two oligonucleotides, one containing a preformed hairpin structure. The yield of the reaction products was maximal at temperatures close to 70°C for either enzyme; their accumulation reached a plateau at 70–75% of the theoretical yield at a stoichiometric enzyme-to-substrate ratio. The enzymes differed in thermal stability. The half-life of Pab DNA ligase was approximately 60 min at 90°C. Mth DNA ligase was completely inactivated within 10 min at this temperature. The recombinant DNA ligases from P. abyssi and M. thermoautotrophicum remained stabile during long-term storage at 4°C.     

Fickean and Non-Fickean Water Desorption During Vacuum Drying of <Emphasis Type="Italic">L. bulgaricus</Emphasis>

The recovery of Lactobacillus bulgaricus was studied in correlation to the kinetics of cell drying. When bacteria were dehydrated at 30 °C, either in the presence or the absence of sucrose, the drying kinetics corresponds to a Fickean diffusion in correspondence with a short lag time. In contrast, when the bacteria were dehydrated at 70 °C in the absence of sugar, the kinetics corresponds to an anomalous diffusion, and the lag time is four to five times higher than that at 30 °C. However, when drying at 70 °C was carried out in the presence of sucrose, drying kinetics turned into a Fickean process parallel to a substantial decrease in the lag time. The pattern of water desorption was correlated with the critical water activity. When the drying kinetics corresponds to a Fickean diffusion, the lag time started to increase at 0.7 water activity, but when the cells were dried at 70 °C, the damage started at 0.5 water activity. This observation indicates that the drying rate affects the pattern of water desorption, and it can change the value of critical water activity. These results put into relevance that the cell recovery is due to the drying history and that the recovery increase produced by sucrose can be related to the maintenance of kinetic barriers for water desorption.     

Enhancement of artemisinin content and biomass in <Emphasis Type="Italic">Artemisia annua</Emphasis> by exogenous GA<Subscript>3</Subscript> treatment

Artemisinin is a promising and potent antimalarial drug naturally produced by the plant Artemisia annua L. but in very low yield. Its artemisinin content is known to be greatly affected by both genotype and environmental factors. In this study, the production of artemisinin and leaf biomass in Artemisia annua L. was significantly increased by exogenous GA₃ treatment. The effect of GA₃ application on expression of proposed key enzymes involved in artemisinin yield was examined in both wild type (007) and FPS-overexpression (253-2) lines of A. annua. In the wild type (007) at 6 h post GA₃ application there was an abrupt rise in FPS, ADS and CYP71AV1 expression and at 24 h a temporary and significant peak in artemisinin (1.45-fold higher than the control). After GA₃ application in line 253-2, there was a dramatic rise in expression of FPS at 3 h, CYP71AV1 at 9 h and ADS at 72 h and accumulation of artemisinin after 7 days, which was a delay when compared with the wild type plant. Thus, increased artemisinin content from exogenous GA₃ treatment was associated with increased expression of key enzymes in the artemisinin biosynthesis pathway. Interestingly, exogenous GA₃ continuously enhanced artemisinin content from the vegetative stage to flower initiation in both plant lines and gave significantly higher leaf biomass than in control plants. Consequently, the artemisinin yield in GA₃-treated plants was much higher than in control plants. Although the maximum artemisinin content was found at the full blooming stage [2.1% dry weight (DW) in 007 and 2.4% DW in 253-2], the highest artemisinin yield in GA₃-treated plants was obtained during the flower initiation stage (2.4 mg/plant in 007 and 2.3 mg/plant in 235-2). This was 26.3 and 27.8% higher, respectively, than in non-treated plants 007 and 253-2. This study showed that exogenous GA₃ treatment enhanced artemisinin production in pot experiments and should be suitable for field application.     

Isozymes of <Emphasis Type="Italic">α</Emphasis>-amylases from newly isolated <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> CKB19: Production from immobilized cells

The aim of this study was to produce two isozymes of α-amylase by immobilization of a newly isolated soil bacterium. The bacterium was identified as Bacillus thuringiensis CKB19 on the basis of its 16S rRNA profile. Enzyme production by free cells increased linearly with cell growth up to 34 h in starch containing enriched liquid media. The active bacterial cells were immobilized in Caalginate beads, and operational stability of the entrapped cell was optimized for amylase production. Enzyme production was optimal at an alginate concentration of 2 g% (w/v), calcium chloride concentration of 1 M, and with 300 beads (each bead contained 2 × 10⁷ cells)/250 mL flask. Amylase production by the immobilized cells was about 3 times higher than free cell fermentation after 34 h of incubation. It was observed that the immobilized bacterium secreted two different amylases (Am-I and Am-II) into the culture fluid. The molecular masses of Am-I and Am-II were 59.6 and 44.7 kd, respectively, and showed optimum activity at pH 5.0 and 9.0. Both amylases showed optimum activity at 40°C and were stable at the same temperature, with losses of only 10 and 20% (for Am I and Am II, respectively) of their original activities after 24 h of incubation. Further, both amylases were salt tolerant (up to 4 M NaCl) and hydrolyzed raw starchy foods into glucose. All these characteristics make this enzyme mixture suitable for use as a digestive aid and for the improvement of digestibility of animal feed ingredients.     

Traitement de l’hyperthyroïdie par l’iode 131 : à propos de 280 cas

We report through this work the experience of nuclear medicine department at Ibn-Sina hospital (Rabat), in the treatment of hyperthyroidism with iodine-131. We retrospectively studied a cohort of 280 patients with hyperthyroidism, from several regions of Morocco between January 2001 and January 2010. A clinical examination and a serum assessment of TSHus, FT4 and FT3 have been made at baseline and at 3, 6, 12 and 24 months after radioiodine therapy. The activity of iodine-131 administered ranged from 296 to 740 MBq and depended on the pathology being treated, age, thyroid volume, intensity of clinical and biological hyperthyroidism and socioeconomic situation. Radioiodine therapy has often been proposed as a treatment for second or third intention, 71 patients were initially treated with iodine-131, while 209 patients had received iodine-131 after failure of medical treatment and/or recurrence after surgical treatment. Graves’ disease was the most common etiology (60%), followed by toxic adenoma (20%), and multinodular toxic goiter (13%). The therapeutic efficacy of a single dose of radioiodine evaluated after a 6-months follow-up was 92% in Graves’ disease, 98% in toxic adenomas and 97% in toxic multinodular goiter. No acute complication was observed. Taking into account our socioeconomic context, radioiodine therapy remains the preferred treatment of hyperthyroidism in our country with good value vs. price and excellent tolerance.     

Complementary expression of<Emphasis Type="Italic"> AP-2</Emphasis> and<Emphasis Type="Italic"> AP-2rep</Emphasis> in ectodermal derivatives of<Emphasis Type="Italic"> Xenopus</Emphasis> embryos

In an attempt to define the pattern of developmental expression of AP-2rep and AP-2 in Xenopus embryos, we cloned a Xenopus AP-2rep cDNA. The AP-2rep message was localized in the organizer region at the gastrula stage whereas AP-2 was expressed ventro-laterally in the animal hemisphere. Later, AP-2rep was expressed in the entire neural tissue at the neurula stage while AP-2 was predominantly expressed in the cranial neural crest areas. The endogenous expression of AP-2 in the neural crest area was diminished by ectopic injection of AP-2rep RNA, suggesting a role for AP-2rep in the differentiation of neural tissues by restricting the expression of AP-2 in the Xenopus embryo.     

Seed production of cut-leaved teasel (<Emphasis Type="Italic">Dipsacus laciniatus</Emphasis>) in central Missouri

Cut-leaved teasel is an invasive weed in Missouri that reduces the diversification of native species along roadsides and impairs traffic visibility. Teasel is a biennial and grows as a rosette in the first year and flowers the second year. Reproduction is only by seed. Field studies were conducted in 2004 and 2005 at two locations to assess the seed production of cut-leaved teasel. From a natural stand, fifteen plants were tagged at the onset of flowering. Selected plants included those considered growing in a group and those growing alone; a plant was considered alone when no other plant was adjacent for at least 60 cm. Whenever a seedhead completed flowering, it was covered with a cellophane bag and harvested one month later. Linear regression was used to correlate the weight of seeds from a single seedhead and number of seeds to estimate the total seed production per seedhead. The number of seedheads per plant varied from 3 to 56. On average, plants growing alone had 64% more seedheads per plant than plants occurring in a group. Seed numbers in the primary seedhead ranged from 511 to 1,487. Total seed production per plant ranged from 1,309 to 33,527. Seed production was 61% greater for plants growing alone versus those growing in a group and was more prolific in 2005 than in 2004. In addition, seed production per plant varied between locations for plants growing alone, but seed yield per plant was similar for plants growing in groups. Colonization of teasel in new areas is facilitated by higher seedhead numbers per plant and total seed production compared to reproduction of plants in areas of intraspecific competition.     

Thermoregulatory physiology of the Crested Pigeon<Emphasis Type="Italic"> Ocyphaps lophotes</Emphasis> and the Brush Bronzewing<Emphasis Type="Italic"> Phaps elegans</Emphasis>

The metabolic physiology of the Crested Pigeon (Ocyphaps lophotes) and the Brush Bronzewing (Phaps elegans) is generally similar to that expected for birds of their size, but the Crested Pigeon has a number of characteristics which would aid survival in hot and dry regions. Body temperature increased similarly for the Crested Pigeon (from 38.8 degrees C to 41.5 degrees C) and the Brush Bronzewing (39.3 degrees C to 41.4 degrees C) over ambient temperatures (T(a)s) from 10 degrees C to 35 degrees C. Both species became hyperthermic (body temperature, T(b)>42 degrees C) at T(a)=45 degrees C. Basal metabolic rate of the Crested Pigeon (0.65 ml O(2) g(-1) h(-1) at 40 degrees C) was approximately 71% of that predicted for a columbid bird, while BMR of the Brush Bronzewing (0.87 ml O(2) g(-1) h(-1) at 20 degrees C to 40 degrees C) was approximately 102% of predicted. Total evaporative water loss increased exponentially with T(a) for both species, from <1 mg H(2)O g(-1) h(-1) at 10 degrees C to >12 mg H(2)O g(-1) h(-1) at 45 degrees C. It was similar and low for both species at T(a)<30 degrees C, but was higher for the Brush Bronzewing than the Crested Pigeon at T(a)>30 degrees C. Ventilatory minute volume matched oxygen consumption, such that oxygen extraction efficiency did not change with T(a) and was similar for both species (approximately 20%). Expired air temperature was considerably lower than T(b) for both species at T(a)<35 degrees C, potentially reducing respiratory water loss by approximately 65% at T(a)=10 degrees C to approximately 30% at T(a)=35 degrees C. Cutaneous evaporative cooling was significant for both species, with skin resistance decreasing as T(a) increased. The Crested Pigeon had a lower skin resistance than the Brush Bronzewing at T(a)=45 degrees C. The Brush Bronzewing had apparently reached its maximum cutaneous water loss at 30 degrees C and relied on panting to cool at higher T(a).     

Accumulation of copper in <Emphasis Type="Italic">Trichoderma reesei</Emphasis> transformants,constructed with the modified <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation technique

A modified Agrobacterium tumefaciens-mediated transformation method was established for the construction of mutants with improved copper tolerance and accumulation capability in Trichoderma reesei. One transformant, AT01, exhibited the highest copper accumulation capability. With copper at 0.7 mM, AT01 removed 13 mg copper/g biomass (removal rate of 96%), whereas the wild-type strain removed only 6 mg copper/g biomass (removal rate of 50%). Optimal conditions were a pH of 5.0 at 28°C. The pigment change of Trichoderma mycelia was a potential indicator of copper accumulation. Electron microscopy revealed that copper was mainly accumulated in cell vacuoles.     

Structural stability and functional analysis of L-asparaginase from <Emphasis Type="Italic">Pyrococcus furiosus</Emphasis>

We report studies on an L-asparaginase from Pyrococcus furiosus, cloned and expressed in Escherichia coli and purified to homogeneity. Protein stability and enzyme kinetic parameters were determined. The enzyme was found to be thermostable, natively dimeric, and glutaminase-free, with optimum activity at pH 9.0. It showed a K _m of 12 mM and a substrate inhibition profile above 20 mM L-asparagine. Urea could not induce unfolding and enzyme inactivation; however, with guanidine hydrochloride (GdnCl) a two-state unfolding pattern was observed. Reduced activity and an altered near-UV-CD signal for protein at low GdnCl concentration (1 M) suggested tertiary structural changes at the enzyme active site. A homology three-dimensional model was developed and the structural information was combined with activity and stability data to give functional clues about the asparaginase.     

Selection perspectives for genetic improvement of wood stiffness in hybrid larch (<Emphasis Type="Italic">Larix</Emphasis> x <Emphasis Type="Italic">eurolepis</Emphasis> Henry)

Genetic variability of modulus of elasticity (MOE) was investigated in three genetic trials, including two progeny (16 years old) and one clonal (19 years old) trials of hybrid larch (Larix x eurolepis Henry). MOE was directly assessed on standing trees using the Rigidimeter, a bending device, and related to other traits including height, BH diameter and wood density. Mean MOE ranged from 5,183 to 9,228 MPa among families in the progeny trials and from 4,591 to 11,486 MPa in the clonal trial. Among traits studied, MOE was one of the most variable traits. It was strongly and positively related to wood density at both the individual and genotype mean levels. Interestingly too, wood stiffness did not seem, or only weakly, unfavourably linked to stem diameter at the phenotypical level, but it was negatively or not correlated to diameter at the genetic level. As well, MOE showed a high GxE stability over the two progeny trial sites. Narrow-sense heritabilities for MOE were moderate (around 0.36). In all three trials, they were lower than those for wood density or total height, and of the same level as for diameter. Improvement of wood stiffness of hybrid larch using the Rigidimeter seems possible and promising genetic gains are expected. Impacts of selection for growth traits on MOE are also discussed.     

Ultrastructure of cyanobacterium <Emphasis Type="Italic">Nostoc</Emphasis> sp. f. <Emphasis Type="Italic">Blasia</Emphasis> cell forms in persisting populations

Cell clusters formed in persistent populations of Nostoc sp. f. Blasia, a cyanobacterium capable of cell differentiation, under prolonged storage in the dark at low temperatures were studied for the first time. Cell reorganization was observed, including changes in the ultrastructure of thylakoids, the cell wall peptidoglycan layer, and carboxysomes. Subcellular structures involved in intercellular communication within the clusters were revealed (structures similar to microplasmodesms and contact pores, secretory vesicles, etc.) Persistence of cyanobacterial populations was concluded to result from formation not only of specialized dormant cells (akinetes), but also L-forms, as well as from the modification changes of the clustered vegetative cells. A cluster containing the vegetative cells and L-like forms within a common intercellular matrix is considered a structural unit at the supracellular level, which is responsible for survival of cyanobacterial populations when mass akinete formation does not occur.     

Hydrolases in supercritical CO<Subscript>2</Subscript> and their use in a high-pressure membrane reactor

The thermal stability and activity of enzymes in supercritical carbon dioxide (SC CO(2)) and near-critical propane were studied at a pressure of 300 bar in the temperature range 20-90 degrees C. Proteinase from Carica papaya was incubated in microaqueous SC CO(2) at atmospheric pressure in a nonaqueous system. Lipase stability in an aqueous medium at atmospheric pressure and in SC CO(2) as well as near-critical propane at 100 bar and 40 degrees C was studied. In order to investigate the impact of solvent on lipases, these were chosen from different sources: Pseudomonas fluorescences, Rhizpous javanicus, Rhizopus niveus and porcine pancreas. On the basis of our previous study on lipase activities in dense gases, a high-pressure continuous flat-shape membrane reactor was designed. The hydrolysis of sunflower oil in SC CO(2) was performed as a model reaction in this reactor. The reaction was catalyzed by the lipase preparation Lipolase 100T and was performed at 50 degrees C and 200 bar.