Epstein-Barr Virus Infection and Altered Control of Apoptotic Pathways in Posttransplant Lymphoproliferative Disorders

Posttransplant lymphoproliferative disorders (PTLD) represent a spectrum of lymphoid diseases complicating the clinical course of transplant recipients. Most PTLD are Epstein-Barr virus (EBV) associated with viral latency type III. Several in vitro studies have revealed an interaction between EBV latency proteins and molecules of the apoptosis pathway. Data on human PTLD regarding an association between Bcl-2 family proteins and EBV are scarce. We analyzed 60 primary PTLD for expression of 8 anti- (Bcl-2, Bcl-XL, and Mcl-1) and proapoptotic proteins (Bak and Bax), the so-called BH3-only proteins (Bad, Bid, Bim, and Puma), as well as the apoptosis effector cleaved PARP by immunohistochemistry. Bim and cleaved PARP were both significantly (p = 0.001 and p = 5.251e-6) downregulated in EBV-positive compared to EBV-negative PTLD [Bim: 6/40 (15%), cleaved PARP: 10/43 (23%), vs. Bim: 13/16 (81%), cleaved PARP: 12/17 (71%)]. Additionally, we observed a tendency toward increased Bcl-2 protein expression (p = 0.24) in EBV-positive PTLD. Hence, we provide evidence of a distinct regulation of Bcl-2 family proteins in EBV-positive versus negative PTLD. The low-expression pattern of the proapoptotic proteins Bim and cleaved PARP together with the high-expression pattern of the antiapoptotic protein Bcl-2 by trend in EBV-positive tumor cells suggests disruption of the apoptotic pathway by EBV in PTLD, promoting survival signals in the host cells.     

Design of a cylindrical brachytherapy implant applicator for the irradiation of an intestinal segment in mice

YingExperimental determination of the RBE of new isotopes for brachytherapy implants (e.g. iodine-125 and palladium-103) remains a very difficult problem, especially in small animals, where the seeds cannot be implanted easily in the planned geometry in a reproducible way. This technical note describes an original device that makes it possible to irradiate a segment of the intestine in mice for the purpose of determining the RBE for crypt regeneration. The device is a length of tube (3.4 mm and 7 mm internal and external diameter, respectively) whose external surface has been longitudinally grooved and into which the seeds can be squeezed (each groove holds either one or two seeds). The tube is composed of two sections. This seed container can be surgically positioned around an intestinal ansa while the mice are anesthetized. The mean dose rates in the intestine (for eight seeds) were found to be 86.3 +/- 5.9 and 79.0 +/- 5.4 cGy/h for 29.2 MBq (1 U) iodine and 28.6 MBq (1 U) palladium seeds, respectively. So far, more than 100 mice have been irradiated successfully. Full dose-effect relationships can be obtained using the same seeds and applying them successively in different groups of animals (which ensured the accuracy of the relative doses).     

Evaluation of the resistance to two nematodes: Radopholus similis and Meloidogyne spp. in four banana genotypes in Morocco

Radopholus similis and Meloidogyne spp. are the main nematode parasites of banana plants grown under plastic shelters in Morocco. A test was made in pots to evaluate the resistance of four genotypes of banana to these nematodes. Infection by Meloidogyne spp. brought about an increase in root weight in all banana plants tested because of gall formation. The inoculation of R. similis produced a reduction in length and diameter of the pseudo-trunk as well as in root and aerial mass in all genotypes. Pisang jari buaya showed the significantly lowest number of Meloidogyne nematodes per 10 g of roots, whereas for R. similis, the significantly smallest numbers were obtained in Pisang berlin and Pisang jari buaya. Therefore, Pisang jari buaya was the only banana genotype studied to show some degree of resistance to both nematodes.     

Use of morphological and physiological characters, and molecular markers to evaluate the genetic diversity of three clementine cultivars

Originating from a natural crossing between mandarin and sweet orange at the end of the 19(th) century, clementine diversified through the selection of spontaneous mutations. Today, it seems almost impossible to distinguish one variety from another. The development of molecular tools for variety identification is thus necessary. Three clementine cultivars, representing distinct groups of fruit maturity, were evaluated. Identification criteria were searched at the phenotypical level (organoleptic characteristics, leaves morphology) as well as the DNA level (isozymes, RAPD, and ISSR). The phenotypical diversity observed is relatively high and contrasted with the low molecular polymorphism. In fact, only the cultivar 'Guerdane' presents profiles of genetic fingerprints different from those of the two other cultivars. The frequency of the genetic modifications would thus be variable from a cultivar to another. Moreover, the specific molecular markers of the cultivar 'Guerdane', added to the phenotypic markers, extend the possibilities of identification to the young nursery plants.     

Use of a homologous radioimmunoassay (RIA) to evaluate the effect of maternal and foetal parameters on pregnancy-associated glycoprotein (PAG) concentrations in sheep

Early pregnancy detection and prediction of the number of lambs would be profitable for sheep breeders because it enables them to adjust nourishment of pregnant ewes according to the individual needs in order to prevent health problems around parturition. The concentration of ovPAG has previously been reported to be related with maternal parameters (farm, breed and age) as well as foetal parameters (number of lambs, their sex and birth weight), but contradictory results were obtained in different small-scale studies. This large-scale study evaluates the effect of these parameters on the ovPAG concentration, determined by a homologous radioimmunoassay (RIA), and it further investigates the possibility to predict the number of lambs by means of homologous ovPAG determination. Eighty-three and ninety-five ewes of the Suffolk and Texel breed, respectively, housed on four different farms (experiment 1) and 68 ewes of the Suffolk breed, housed on two different farms (experiment 2) were included in this study, and their estrous cycles were synchronised using a progesterone analogue. On the day of synchronisation (D-14) and at 25 (D25), 35 (D35) and 45 (D45) days after insemination, blood samples were taken and a homologous radioimmunoassay (RIA) was used to determine the ovPAG concentrations. At parturition, age of the ewe, number and sex of the lambs (experiment 1) and birth weight (experiment 2) were registered. OvPAG concentrations were not affected by age of the ewe and sex of the lambs. Farm and breed of the ewes, number and birth weight of the lambs had a significant effect on ovPAG concentrations at all time points (P<0.05). The odds of multiple lambs increased significantly with increasing ovPAG concentration, although prediction of litter size based on ovPAG concentration at the individual ewe level was not useful due to small sensitivity and/or specificity whatever the cutoff value used. In conclusion, the ovPAG concentration is affected by farm and breed of the ewes, and number and birth weight of the lambs.     

Biological and mathematical modeling of melanocyte development

We aim to evaluate environmental and genetic effects on the expansion/proliferation of committed single cells during embryonic development, using melanoblasts as a paradigm to model this phenomenon. Melanoblasts are a specific type of cell that display extensive cellular proliferation during development. However, the events controlling melanoblast expansion are still poorly understood due to insufficient knowledge concerning their number and distribution in the various skin compartments. We show that melanoblast expansion is tightly controlled both spatially and temporally, with little variation between embryos. We established a mathematical model reflecting the main cellular mechanisms involved in melanoblast expansion, including proliferation and migration from the dermis to epidermis. In association with biological information, the model allows the calculation of doubling times for melanoblasts, revealing that dermal and epidermal melanoblasts have short but different doubling times. Moreover, the number of trunk founder melanoblasts at E8.5 was estimated to be 16, a population impossible to count by classical biological approaches. We also assessed the importance of the genetic background by studying gain- and loss-of-function β-catenin mutants in the melanocyte lineage. We found that any alteration of β-catenin activity, whether positive or negative, reduced both dermal and epidermal melanoblast proliferation. Finally, we determined that the pool of dermal melanoblasts remains constant in wild-type and mutant embryos during development, implying that specific control mechanisms associated with cell division ensure half of the cells at each cell division to migrate from the dermis to the epidermis. Modeling melanoblast expansion revealed novel links between cell division, cell localization within the embryo and appropriate feedback control through β-catenin.     

Gene expression between a congenic strain that contains a quantitative trait locus of high bone density from CAST/EiJ and its wild-type strain C57BL/6J

Peak bone density is an important determining factor of future osteoporosis risk. We previously identified a quantitative trait locus (QTL) that contributes significantly to high bone density on mouse chromosome 1 from a cross between C57BL/6J (B6) and CAST/EiJ (CAST) mouse strains. We then generated a congenic strain, B6.CAST-1T, in which the chromosomal fragment containing this QTL had been transferred from CAST to the B6 background. The congenic mice have a significantly higher bone density than the B6 mice. In this study we performed cDNA microarray analysis to evaluate the gene expression profile that might yield insights into the mechanisms controlling the high bone density by this QTL. This study led to several interesting observations. First, approximately 60% of 8,734 gene accessions on GEM I chips were expressed in the femur of B6 mice. The expression and function of two-thirds of these expressed genes and ESTs have not been documented previously. Second, expression levels of genes related to bone formation were lower in congenic than in B6 mice. These data are consistent with a low bone formation in the congenic mice, a possibility that is confirmed by reduced skeletal alkaline phosphatase activity in serum compared with B6 mice. Third, expression levels of genes that might have negative regulatory action on bone resorption were higher in congenic than in B6 mice. Together these findings suggest that the congenic mice might have a lower bone turnover rate than B6 mice and raise the possibility that the high bone density in the congenic mice could be due to reduced bone resorption rather than increased bone formation. Electronic Publication     

Plant genomics in Africa: present and prospects

Plants are the world’s most consumed goods. They are of high economic value and bring many health benefits. In most countries in Africa, the supply and quality of food will rise to meet the growing population’s increasing demand. Genomics and other biotechnology tools offer the opportunity to improve subsistence crops and medicinal herbs in the continent. Significant advances have been made in plant genomics, which have enhanced our knowledge of the molecular processes underlying both plant quality and yield. The sequencing of complex genomes of African plant species, facilitated by the continuously evolving next-generation sequencing technologies and advanced bioinformatics approaches, has provided new opportunities for crop improvement. This review summarizes the achievements of genome sequencing projects of endemic African plants in the last two decades. We also present perspectives and challenges for future plant genomic studies that will accelerate important plant breeding programs for African communities. These challenges include a lack of basic facilities, a lack of sequencing and bioinformatics facilities, and a lack of skills to design genomics studies. However, it is imperative to state that African countries have become key players in the plant genome revolution and genome derived-biotechnology. Therefore, African governments should invest in public plant genomics research and applications, establish bioinformatics platforms and training programs, and stimulate university and industry partnerships to fully deploy plant genomics, particularly in the fields of agriculture and medicine.     

A novel role for central ACBP/DBI as a regulator of long‐chain fatty acid metabolism in astrocytes

Acyl‐CoA‐binding protein (ACBP) is a ubiquitously expressed protein that binds intracellular acyl‐CoA esters. Several studies have suggested that ACBP acts as an acyl‐CoA pool former and regulates long‐chain fatty acids (LCFA) metabolism in peripheral tissues. In the brain, ACBP is known as Diazepam‐Binding Inhibitor, a secreted peptide acting as an allosteric modulator of the GABA_A receptor. However, its role in central LCFA metabolism remains unknown. In the present study, we investigated ACBP cellular expression, ACBP regulation of LCFA intracellular metabolism, FA profile, and FA metabolism‐related gene expression using ACBP‐deficient and control mice. ACBP was mainly found in astrocytes with high expression levels in the mediobasal hypothalamus. We demonstrate that ACBP deficiency alters the central LCFA‐CoA profile and impairs unsaturated (oleate, linolenate) but not saturated (palmitate, stearate) LCFA metabolic fluxes in hypothalamic slices and astrocyte cultures. In addition, lack of ACBP differently affects the expression of genes involved in FA metabolism in cortical versus hypothalamic astrocytes. Finally, ACBP deficiency increases FA content and impairs their release in response to palmitate in hypothalamic astrocytes. Collectively, these findings reveal for the first time that central ACBP acts as a regulator of LCFA intracellular metabolism in astrocytes.

     

Caffeoyl coenzyme A O-methyltransferase down-regulation is associated with modifications in lignin and cell-wall architecture in flax secondary xylem

Caffeoyl coenzyme A O-methyltransferase (CCoAOMT, EC 2.1.1.104) down-regulated-flax (Linum usitatissimum) plants were generated using an antisense strategy and functionally characterized. Chemical analyses (acetyl bromide and thioacidolysis) revealed that the lignin quantity was reduced and that the Syringyl/Guaïacyl (S/G) lignin monomer ratio was modified in the non-condensed lignin fraction of two independent down-regulated lines. These modifications were associated with altered xylem organization (both lines), reduced cell-wall thickness (one line) and the appearance of an irregular xylem (irx) phenotype (both lines). In addition UV microspectroscopy also indicated that CCoAOMT down-regulation induced changes in xylem cell-wall structure and the lignin fractions. Microscopic examination also suggested that CCoAOMT down-regulation could influence individual xylem cell size and identity. As a first step towards investigating the cellular mechanisms responsible for the unusual structure of flax lignin (G-rich, condensed), recombinant flax CCoAOMT protein was produced and its affinity for different potential substrates evaluated. Results indicated that the preferred substrate was caffeoyl coenzyme A, followed by 5-hydroxyconiferaldehyde suggesting that flax CCoAOMT possesses a small, but probably significant 5′ methylating activity, in addition to a more usual 3′ methylating activity.