A new transformation-regeneration procedure in the model legume Lotus japonicus: root explants as a source of large numbers of cells susceptible to Agrobacterium-mediated transformation

We describe herein a simple and efficient transformation procedure for the production of transgenic Lotus japonicus plants. In this new procedure, dedifferentiated root explants, used as starting material, are the source of a large number of cells that are competent for the regeneration procedure, with a high susceptibility to Agrobacterium infection. The application of this protocol resulted in a tenfold increase in the number of transformants produced by a single plant in comparison to the widely used hypocotyl transformation procedure. Furthermore, our procedure allowed the use of intact plants stored for a long time at 4 degrees C, thus providing a potential continuous supply of explants for transformation experiments. The overall time of incubation under tissue culture conditions required to obtain a plant transferable into soil is 4 months. The transgenic nature of the transformants was demonstrated by the detection of beta-glucuronidase (GUS) activity in the primary transformants and by molecular analysis. Stable transformation was indicated by Mendelian segregation of the hygromycin selectable marker and of the gusA activity after selfing of the transgenic plants.     

Biochemical composition of crustacean zooplankton and their grazing on phytoplankton and ciliated protozoans in a recently founded reservoir (Sahela, Morocco

In order to assess the impact of crustacean zooplankton on phytoplankton and protozoan ciliates in the Sahela reservoir under semi-arid climate, we conducted experiments during the period from July to December 1999 at the deepest point in the lake (15 m). Samplings and measurements were carried out in diffusion chambers submerged in situ over a period of 7 h without (control chambers) and with (experimental chambers) crustacean zooplankton. During these experiments, counts were conducted on phytoplankton and ciliates to determine the abundance and the mortality of these organisms due to zooplankton in each diffusion chambers at t = 0 and t = 7 h of incubation. The study showed that the growth rates of phytoplankton and ciliates populations varied between 0.02 and 0.05 h-1 and from 0.01 to 0.07 h-1, respectively. The mortality caused by zooplankton grazing fluctuated from 0.07 to 0.2 h-1 of phytoplankton and from 0.01 to 0.2 h-1 of ciliates. These mortalities were significantly and positively correlated with the growth rates (r = 0.8; p < 0.02; n = 9). Moreover, the heavy predation by the crustacean zooplankton was exerted on small-sized phytoplankton and ciliates and we demonstrated the relationships between protozoans and zooplankton for the transfer of matter and energy in aquatic food webs. Furthermore, the crustacean zooplankton metabolism was different, whether zooplankton was present in diffusion chambers or in the lake.     

Stability and Hopf bifurcation in an approachable haematopoietic stem cells model

We consider the haematopoietic stem cells model (HSC) with one delay introduced by Mackey [M.C. Mackey, Unified hypothesis for the origin of aplastic anemia and periodic hematopoiesis, blood 51 (1978) 5; M.C. Mackey, Mathematical models of haematopoietic cell replication and control, in: The Art of Mathematical Modelling: Case Studies in Ecology, Physiology and Biofluids, H.G. Othmer, F.R. Adler, M.A. Lewis, J.C. Dallon (Eds), Prentice-Hall, New York, 1997, p. 149] and Andersen and Mackey [L.K. Andersen, M.C. Mackey, Resonance in periodic chemotherapy: a case study of acute myelogenous leukemia, J. theor. Biol. 209 (2001) 113]. There are two possible stationary states in the model. One of them is trivial and the second E( *)(tau) depending on the delay is non-trivial . This paper investigates the stability of the non-trivial state and occurrence of the Hopf bifurcation depending on time delay. We prove the existence and uniqueness of a critical values tau(0) and tau of the delay such that E( *)(tau) is asymptotically stable for tau相似文献   

Protein-bound polyamines in the plasma of mice grafted with the Lewis lung carcinoma

Protein-bound polyamines were isolated from the plasma of mice using antipolyamine antibodies covalently linked to magnetic latex spheres. Their subsequent separation by polyacrylamide gel electrophoresis (PAGE) showed that in plasma from normal mice, 3 proteins (27, 55 and 82 kDa) carrying polyamines could be visualized, whereas in mice bearing the Lewis lung carcinoma at least 8 other proteins of higher molecular mass (5 of 94, 110, 130, 145 and 160 kDa, and 3 of greater than 170 kDa) had bound polyamines. These protein-bound polyamines could be detected from the first week after tumour graft; they increased during the second and third week but decreased thereafter. These proteins were not bound by immunolatex spheres preincubated with spermine bound to a protein-carrier insulin. Moreover, the appearance of these protein-bound polyamines was not a consequence of the inflammatory process since in mice infected with heat-inactivated Brucella abortus, with the exception of a 65 kDa protein, polyamines were bound to the same proteins found in normal mice. In mice grafted with the Lewis lung carcinoma the concomitant decrease in transglutaminase-mediated polyamine (e.g. putrescine) binding capacity of plasma proteins provides additional evidence for the presence in vivo of polyamines already bound to plasma proteins.     

Genetic Variation in the Interleukin-28B Gene Is Associated with Spontaneous Clearance and Progression of Hepatitis C Virus in Moroccan Patients

Background

Genetic variation in the IL28B gene has been strongly associated with treatment outcomes, spontaneous clearance and progression of the hepatitis C virus infection (HCV). The aim of the present study was to investigate the role of polymorphisms at this locus with progression and outcome of HCV infection in a Moroccan population.

Methods

We analyzed a cohort of 438 individuals among them 232 patients with persistent HCV infection, of whom 115 patients had mild chronic hepatitis and 117 had advanced liver disease (cirrhosis and hepatocellular carcinoma), 68 individuals who had naturally cleared HCV and 138 healthy subjects. The IL28B SNPs rs12979860 and rs8099917 were genotyped using a TaqMan 5′ allelic discrimination assay.

Results

The protective rs12979860-C and rs8099917-T alleles were more common in subjects with spontaneous clearance (77.9% vs 55.2%; p = 0.00001 and 95.6% vs 83.2%; p = 0.0025, respectively). Individuals with clearance were 4.69 (95% CI, 1.99–11.07) times more likely to have the C/C genotype for rs12979860 polymorphism (p = 0.0017) and 3.55 (95% CI, 0.19–66.89) times more likely to have the T/T genotype at rs8099917. Patients with advanced liver disease carried the rs12979860-T/T genotype more frequently than patients with mild chronic hepatitis C (OR = 1.89; 95% CI, 0.99–3.61; p = 0.0532) and this risk was even more pronounced when we compared them with healthy controls (OR = 4.27; 95% CI, 2.08–8.76; p = 0.0005). The rs8099917-G allele was also associated with advanced liver disease (OR = 2.34; 95% CI, 1.40–3.93; p = 0.0100).

Conclusions

In the Moroccan population, polymorphisms near the IL28B gene play a role both in spontaneous clearance and progression of HCV infection.     

Abundance and biomass of rotifers in relation to the environmental factors in geothermal waters in Southern Tunisia

The spatial and temporal dynamics of rotifers in relation to the physico-chemical parameters in Fish-Culture Research Station (Southern Tunisia) were studied monthly from February 2005 to January 2006. Thirteen rotifer species were found: Brachionus urceolaris, Brachionus calyciflorus, Brachionus sp., Lecane stichaea, Lecane rhytida, Lecane sp., Hexarthra mira, Rotaria tardigrada, Conochiloides natans, Trichocerca marina, Keratella quadrata, Keratella cochlearis and Notommata codonella. The most dominant rotifer was B. urceolaris (76% of total abundance). Rotifer density and water temperature were negatively correlated (r=−0.94, n=12, p=0.001). The highest abundance of rotifers was found in basin 4 (1.5×10⁵ ind m⁻³, in June 2005).     

Recent dynamics of the wet pastures at Oukaimeden plateau (High Atlas mountains, Morroco)

Wetlands are a very important universal heritage; in the Moroccan High Atlas mountains the geomorphology and the slopes evolution determine the existence of the numerous projecting ledges of pozzines on plateaus and oozing. This is particularly the case in Oukaimeden site located at 75 km south of Marrakesh. These wet grasslands have been developed on small surface areas supported by soils water accumulation; and close to the sources and on some banks constitute wetlands of high floristic richness, which makes them very coveted pastures by local populations. These wet pastures are highly sensitive to climatic factors. Recent climatic changes, the anthropic and pastoral pressures and the global environmental changes, affect these zones at the level of their structure (areas, soil characteristics…) and their ecological functions. In the Oukaimeden plateau, the use of aerial photography made it possible to carry out a detailed follow up of the hydraulic adjustments (dam construction in the 1970s) as well as the setting up of new adjustments (tracks, paths, drain channel) on the structure and functioning of these grasslands.     

Possible pathogenic role of the transmembrane isoform of CD160 NK lymphocyte receptor in paroxysmal nocturnal hemoglobinuria

PIGA mutations in paroxysmal nocturnal hemoglobinuria (PNH) patients lead to a glycosylphosphatidylinositol (GPI)-linked membrane proteins expression deficiency. Herein, we report the constitutive expression of the transmembrane CD160 (CD160-TM) activating receptor on non PIGA-mutated PNH patients circulating NK cells. In healthy individuals, only the GPI-anchored isoform of CD160 receptors is expressed on the circulating NK lymphocytes, while the transmembrane isoform appears after ex vivo activation. Similarly to CD160-GPI, we identified CD160-TM as a receptor for the MHC class I molecules. We demonstrate that PNH patients NK lymphocytes spontaneously produce significant amounts of IFN-γ that is inhibited by anti-CD160-TM or anti-MHC class I mAbs. These results indicate that circulating NK cells from PNH patients exhibit a self-MHC class I molecule reactive effector function, which could be mediated through the recruitment of CD160-TM receptor. Our data provide new insights regarding the possible role of CD160-TM on PNH patients NK lymphocytes and in the pathogenesis of the disease.     

Further SAR studies on novel small molecule inhibitors of the hepatitis C (HCV) NS5B polymerase

Herein, we describe the structure-activity relationship (SAR) of N,N-disubstituted phenylalanine series of NS5B polymerase inhibitors of hepatitis C. The NS5B polymerase inhibitory activity of the most active compound exhibited an IC(50) of 2.7 microM.