Impact of immunosuppression on the development of Epstein-Barr virus (EBV) viremia after pediatric liver transplantation.

OBJECTIVES: Pediatric liver transplant (OLT) patients are at risk of posttransplant lymphoproliferative disease (PTLD) from Epstein-Barr virus (EBV). This study examined the impact of induction and immunosuppression on EBV viremia. METHODS: A retrospective chart review was performed on 197 pediatric patients and induction regimen, immunosuppression levels, and EBV viremia were documented for 1 year post-OLT. Logistic regression models determined associations between induction, immunosuppression, and EBV. RESULTS: Fifty six percent of patients developed EBV viremia. Incidence of EBV viremia was 73% with antithymocyte globulin (ATG), 63% with daclizumab, and 39% for neither, though the trend was not significant [ATG: odds ratio (OR) 0.19; 95% confidence interval (CI) 0.024-1.58; P = .125; daclizumab OR; 1.07; 95% CI 0.270-4.23; P = .925]. Tacrolimus immunosuppression levels were supratherapeutic 28.7% of the time; however, only supratherapeutic tacrolimus levels between 0 and 2 weeks increased EBV viremia at 2 to 4 weeks post-OLT (OR 1.80; 95% CI 1.10-2.94; P = .02). Three patients developed PTLD. CONCLUSIONS: The use of ATG and daclizumab induction likely does not play a role in the development of EBV viremia. Supratherapeutic tacrolimus levels 0 to 2 weeks post-OLT impact the development of EBV viremia at 2 to 4 weeks. The incidence of PTLD was low, suggesting better EBV and immunosuppression monitoring plays an important role in reducing PTLD.

Genetic diversity in the northernmost Oryza rufipogon populations estimated by SSR markers.

To estimate genetic diversity of the residual northern populations of Oryza rufipogon, a total of 232 individuals from six populations were analyzed using microsatellites (SSRs). The O. rufipogon populations with different status included three from Dongxiang (Jiangxi Province) and three from Chaling (Hunan Province) in China. The 23 rice SSR primer pairs selected from the RiceGenes Database detected a total of 115 alleles, indicating that all the SSR loci were polymorphic in this study. The total gene diversity was 0.919 in the six O. rufipogon populations, and the Donxiang populations showed higher diversity than the Chaling populations. More significant genetic differentiation and less gene flow were found among the Dongxiang populations than those from Chaling. The two putative introgressed populations showed relatively high genetic variation. One in situ conserved population from Dongxiang had the lowest level of genetic diversity. The re-introduced population from Chaling restored about 90% of the genetic variation, compared with the original source population. It is concluded from these results that a relatively high level of genetic variation resided in the northern O. rufipogon populations and continued efforts of conservation of these populations are needed; and that the conservation of some Chaling and Dongxiang populations has been effective in preventing gene flow from cultivated rice. Introgression of cultivated rice demonstrated significant impacts on genetic variability of the O. rufipogon populations, and should be carefully considered in conserving this wild rice. This study also suggested that re-introduction to its original habitats is an effective approach to restore O. rufipogon populations.

Genetic differentiation of wild relatives of rice as assessed by RFLP analysis.

To study genetic diversity and relationships of wild relatives of rice, 58 accessions of Oryza rufipogon, Oryza nivara, Oryza sativa f. spontanea and the cultivated Oryza sativa, representing a wide range of their distribution, were analyzed using the restriction fragment length polymorphism (RFLP) technique. All 30-used RFLP probes detected polymorphisms among the Oryza accessions, with an average of 3.8 polymorphic fragments per probe. Considerable genetic diversity was scored among the Oryza accessions with a similarity coefficient ranging from 0.28 to 0.93; but the cluster analysis of the accessions did not show an apparent grouping based on the species classification, instead they were scattered randomly in different groups. Noticeably, the Oryza accessions from the same geographic region, or near-by geographic regions, tended to be clustered in the same groups. The indica rice varieties showed relatively high genetic diversity and were scattered in different groups of their wild relatives, but the japonica varieties showed a relatively low variation and formed an independent group. It is concluded from the molecular analytical result that: (1) the four Oryza taxa have a remarkably close relationship and their independent species status need to be carefully reviewed; (2) geographic isolation has played a significant role in the differentiation of the Oryza accessions; therefore, a wide geographic range needs to be covered in collecting wild rice germplasm for ex situ conservation; and (3) the conventional conclusion of indica rice being directly domesticated from its ancestral wild species, and japonica rice being derived from indica, gains support from our data.

Sulfoglycolipid from the marine brown alga Sargassum hemiphylum.

One kinds of glycolipid (SBI) have been isolated from the marine brown alga Sargassum hemiphyllum (Turn.) Ag. The structures of SBI have been determined as the sodium salt of 1-0-acyl-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol (acyl: tetradecanoyl, pentadecanoyl, 11-hexadecenoyl, hexadecanoyl, 10,13-octadecadienoyl, 9-octade cenoyl, 15-metylheptadecanoyl and 11-eicosenoyl 17: 1.5: 19: 153: 1: 19: 1: 2) on the basis of chemical and spectral evidence and GC-MS analysis, respectively. Four constituents of the SBI were new compounds [the sodium salt of 1-0-(11"-hexadecenoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol, the sodium salt of 1-0-(10",13"-octadecadienoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol, and the sodium salt of 1-0-(15"-metylhexadecenoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol, and the sodium salt of 1-0-(11"-eicosenoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol]. All compounds were isolated from marine brown alga for the first time.

Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes.

The rice genus (Oryza L.) consists of 24 species with 10 recognized genome types. With the realization of many useful genes in species of wild rice, continuous efforts have been made to understand their genomic composition and relationships. However, the identification of rice genomes has often been difficult owing to complex morphological variation and formation of allotetraploids. Here we propose a rapid and reliable method for identifying rice genomes based on the restriction sites of PCR-amplified Adh genes. The experimental procedure was as follows: (i) amplify a portion of Adh1 and Adh2 genes with the locus-specific PCR primers; (ii) digest PCR products with restriction enzymes that distinguish different genomes; and (iii) run the digested products on 1.4% agarose gel, and photograph. Using various combinations of restriction digestion of the two Adh genes, all of the rice genomes can be identified.

Phylogeny of rice genomes with emphasis on origins of allotetraploid species.

The rice genus, Oryza, which comprises 23 species and 9 recognized genome types, represents an enormous gene pool for genetic improvement of rice cultivars. Clarification of phylogenetic relationships of rice genomes is critical for effective utilization of the wild rice germ plasm. By generating and comparing two nuclear gene (Adh1 and Adh2) trees and a chloroplast gene (matK) tree of all rice species, phylogenetic relationships among the rice genomes were inferred. Origins of the allotetraploid species, which constitute more than one-third of rice species diversity, were reconstructed based on the Adh gene phylogenies. Genome types of the maternal parents of allotetraploid species were determined based on the matK gene tree. The phylogenetic reconstruction largely supports the previous recognition of rice genomes. It further revealed that the EE genome species is most closely related to the DD genome progenitor that gave rise to the CCDD genome. Three species of the CCDD genome may have originated through a single hybridization event, and their maternal parent had the CC genome. The BBCC genome species had different origins, and their maternal parents had either a BB or CC genome. An additional genome type, HHKK, was recognized for Oryza schlechteri and Porteresia coarctata, suggesting that P. coarctata is an Oryza species. The AA genome lineage, which contains cultivated rice, is a recently diverged and rapidly radiated lineage within the rice genus.

Biosystematic study of hexaploids Elymus tschimganicus and E. glaucissimus. I. Morphology and genomic constitution.

Elymus tschimganicus and E. glaucissimus are hexaploids (2n = 6x = 42) that inhabit stony slopes and swales in the middle and upper mountain belts of Central Asia. Chromosome pairing at metaphase I in E. tschimganicus and E. glaucissimus was typical of other Elymus hexaploids, averaging 20.94 and 20.98 bivalents per cell, respectively. Meiotic pairing of tetraploid hybrids derived by crossing E. tschimganicus and E. glaucissimus with P. spicata (2n = 2x = 14; genome = SS) averaged 4.97 and 4.90 bivalents per cell respectively, indicating the presence of the S genome in the target taxa. Pentaploid hybrids derived by crossing E. tschimganicus and E. glaucissimus with tetraploid analyzer species E. mutabilis and E. sibiricus (2n = 4x = 28; genome = SSHH) had mean bivalent frequencies of 5.61 and 4.64 respectively, suggesting the absence of the H genome in the target taxa. Mean bivalent associations were similar among hybrids derived by crossing the target taxa with E. nevski (2n = 4x = 28; genome = SSYY), which averaged 10.61 and 11.42 bivalents, respectively, suggesting the presence of the Y genome in E. tschimganicus and E. glaucissimus. Based on chromosome pairing in the hybrids, the genomic formula for both E. tschimganicus and E. glaucissimus should be SSSSYY.

Biosystematic study of hexaploids Elymus tschimganicus and E. glaucissimus. II. Interspecific hybridization and genomic relationship.

To investigate genomic relationships of Elymus tschimganicus (Drobov) Tzvelev (2n = 6x = 42, S1S2Y genomes) and E. glaucissimus (M. Pop.) Tzvelev (2n = 6x = 42, S1S2Y genomes), interspecific hybridizations of the two target species were carried out with 27 other Elymus species containing the SH, SY, SYH, SYP, SYW, and SH1H2 genomes, respectively, collected from different geographic regions. Chromosome pairing behavior was analyzed at metaphase I in 27 hybrids representing 23 hybrid combinations, and overall genomic relationships of the two target species with the other Elymus taxa were estimated. The study concluded that (i) interspecific hybridization was principally easy to perform between the Elymus species, but no general pattern of crossability was obtained, and all hybrids were completely sterile, (ii) the two species have a similar meiotic pattern in their hybrids with the other Elymus species, and (iii) species containing the SY, SYP, and SYH genomes have a generally higher level of genomic homology to the target species than those possessing the SH genomes, and the South American hexaploid with the SH1H2 genomes has the lowest level of genomic homology to the two target taxa.

Genomic constitutions of four Chinese endemic Elymus species: E. brevipes, E. yangii, E. anthosachnoides, and E. altissimus (Triticeae, Poaceae).

The objectives of this study were to determine the genomic constitution and to explore the genomic variation within four Chinese endemic Elymus species, i.e., E. brevipes (Keng) Löve (2n = 4x = 28) and E. yangii B.R. Lu (2n = 4x = 28), E. anthosachnoides (Keng) Löve (2n = 4x = 28), and E. altissimus (Keng) Löve (2n = 4x = 28). Intraspecific crosses between different populations of the four Elymus species, as well as interspecific hybridizations among the four target species, and with six analyzer species containing well-known genomes, i.e., E. caninus (L.) L. (2n = 4x = 28, SH), E. sibiricus L. (2n = 4x = 28, SH), E. semicostatus (Lees ex Steud.) Melderis (2n = 4x = 28, SY), E. parviglumis (Keng) Löve (2n = 4x = 28, SY), E. tsukushiensis Honda (2n = 6x = 42, SHY), and E. himalayanus (Nevski) Tzvelev (2n = 6x = 42, SHY), were achieved through the aid of embryo rescue. Chromosome pairing behaviors were studied in the parental species and their hybrids. Numerical analysis on chromosome pairing was made on the interspecific hybrids. With one exception, each meiotic configuration at metaphase I in the hybrids involving the target taxa and the analyzer species containing the "SH" genomes fit a 2:1:1 model with x-values ranging between 0.91 and 1.00; chromosome pairing in the hybrids involving analyzer parents with the "SY" genomes match a 2:2 model, with x-values between 0.97 and 0.99. All pentaploid hybrids with a genomic formula "SSYYH," except for two crosses having unexpected low c-values, had pairing patterns fitting the 2:2:1 model with x-values varying between 0.96 and 1.00. It is concluded based on hybridization, fertility, and chromosome pairing data that (i) the four target Elymus species are strictly allotetraploid taxa, (ii) they are closely related species, all comprised of the "SY" genomes, (iii) minor genomic structural rearrangements have occurred within the four Elymus species, and (iv) meiotic pairing regulator(s) exists in some of the Elymus taxa studied.

Dihaploids of Elymus from the interspecific crosses E. dolichatherus x E. tibeticus and E. brevipes x E. panormitanus.

Dihaploids (n=2x=14, SY) of two Elymus species, i.e., E. dolichatherus (Keng) Löve (2n=4x=28, SSYY) and E. brevipes (Keng) Löve (2n=4x=28, SSYY), were obtained from the interspecific hybrid combinations E. dolichatherus (♀) x E. tibeticus (Meld.) G. Singh (♂) and E. brevipes (♀) x E. panormitanus (Parl.) Tzvelev (♂). The dihaploids were probably formed through selective elimination of male parental chromosomes in early embryo development. Meiotic chromosome behavior was studied in E. dolichatherus, E. brevipes, and their dihaploids. The two parental Elymus species had regular meioses with predominantly ring bivalent formation. A low frequency of homoeologous chromosome pairing was observed, with an average of 0.81 bivalents and 0.03 trivalents in the dihaploid of E. dolichatherus, and 0.26 bivalents in the dihaploid of E. brevipes. Up to two chromatid bridges accompanied by small fragments were present at anaphase I of the E. dolichatherus dihaploid. It is concluded from this study that: (i) both E. dolichatherus and E. brevipes are allotetraploid species; (ii) a low affinity exists between the "S" and "Y" genomes of the two Elymus species.

Cytogenetic studies of the intergeneric hybrids between Secale cereale and Elymus caninus, E. brevipes, and E. tsukushiensis (Triticeae: Poaceae).

Integeneric hybridizations were carried out between Secale cereale L. (2n = 14, RR) and three Elymus species, namely, E. caninus (L.) L. (2n = 28, SSHH), E. brevipes (Keng) Löve (2n = 28, SSYY) and E. tsukushiensis Honda (2n = 42, SSHHYY). Chromosome pairing was studied at metaphase I in the parental species and the hybrids. Meiotic configurations of the hybrids were 20.74 1+0.14 II for E. caninus x S. cereale (SHR), 16.35 I+2.17 II+0.09 III for E. brevipes x S. cereale (SYR) and 25.84 I+1.10 II+0.02 III for E. tsukushiensis x S. cereale (SHYR), in addition to some secondary associations in the different hybrids. It is concluded from the study that (1) a certain, different homoeologous relationship exists among "S", "H" and "Y" genomes in the investigated Elymus species; (2) low homoeology is present between genomes of Elymus (S or H or Y) and rye (R); (3) the Secale genome affects homoeologous chromosome pairing between different genomes in E. brevipes and E. tsukushiensis.