Post-transplant persistent lymphopenia is a strong predictor of late survival in isolated intestine and multivisceral transplantation.

Absolute lymphocyte count (ALC) has been identified as a prognostic factor in liver transplantation. We hypothesized that a lower ALC may be linked to poor outcomes in isolated intestinal/multivisceral transplantation (IIT/MVT). The aim of this study was to investigate the prognostic impact of ALC in IIT/MVT. A total 141 IIT/MVT patients were eligible for the study. Post-transplant ALCs (at 3, 6, and 12 months) were evaluated, and prognostic impact of trend of ALC during the first year was investigated. Of these 141 patients, 108 patients survived in the first year (1-year survivors). One-year survivors were categorized according to post-transplant ALC at each time point. When ALC was decreased throughout the first year (post-transplant persistent lymphopenia: <500/µl at 3, 6, and 12 months), patient survival (P < 0.001, hazard ratio = 5.09) and graft survival (P < 0.001, hazard ratio = 5.15) after the first year was significantly worse, and this remained to be an independent risk factor. Negative impact of persistent lymphopenia on patient and graft survival was significant regardless of type of intestinal graft. Infection leading to mortality occurred more frequently in the persistent lymphopenia group (43% vs. 24%). Trend of post-transplant ALC may be a strong predictive marker for long-term outcome in 1-year survivors after IIT/MVT.

Sequencing arrays for screening multiple genes associated with early-onset human retinal degenerations on a high-throughput platform.

PURPOSE: To develop and apply microarray-based resequencing technology to detect sequence alterations in multiple autosomal recessive retinal disease genes on a single high-throughput platform. METHODS: Oligonucleotides corresponding to both strands of the target exons and the flanking intron sequences of 29,214 bp from 11 genes associated with autosomal recessive retinitis pigmentosa (arRP) were tiled on 20 x 25-microm microarrays (arRP-I arrays). A total of 155 exons were amplified from 35 arRP patient DNA samples, with each sample being sequenced on an arRP-I chip by hybridization. RESULTS: With the arRP-I arrays, 97.6% of the tiled sequence were determined with more than 99% accuracy and reproducibility. Of the 2.4% unread sequence, 89.5% involved stretches of G or C. In analyzing the 903,140-bp sequence from the 35 patient samples, 506 sequence changes have been detected in which 386 are previously reported alterations, and 120 are novel. In addition to four known causative mutations, six novel sequence changes that are potentially pathogenic were observed. Additional analysis is needed to determine whether these changes are responsible for arRP in these patients. CONCLUSIONS: The use of microarray for sequencing is a novel approach, and the arRP-I chip is the first successful application of this technology for determining sequence alteration in multiple disease-related genes. These arrays can be used for high-throughput genotyping of patients with relevant retinal conditions. In addition, these arrays offer a unique opportunity to interrogate complex patterns of inheritance due to the involvement of more than one gene by screening multiple genes on a single platform.

Late-onset autosomal dominant macular dystrophy with choroidal neovascularization and nonexudative maculopathy associated with mutation in the RDS gene.

PURPOSE: To examine the molecular genetic basis and phenotypic characteristics of an unusual late-onset autosomal dominant macular dystrophy with features of age-related macular degeneration (AMD) in a large family (SUNY901), by using linkage and mutation analyses. METHODS: Blood samples were collected from 17 affected members, 17 clinically unaffected members, and 5 unrelated spouses. Clinical analyses included a review of medical history and standard ophthalmic examination with fundus photography, fluorescein angiography, and electroretinography. Linkage and haplotype analyses were performed with microsatellite markers. Mutation analysis was performed by amplification of exons followed by sequencing. RESULTS: A wide spectrum of clinical phenotypes including exudative and nonexudative maculopathy was observed, with onset in the late fifth decade. Linkage analysis excluded most of the previously known maculopathy loci. Markers D6S1604 (Z(max) of 3.18 at theta = 0), and D6S282 (Z(max) of 3.18 at theta = 0) gave significant positive LOD scores and haplotype analysis localized the disease gene to a 9-centimorgan (cM) interval between markers D6S1616 and D6S459. Mutation analysis excluded the GUCA1A and GUCA1B genes and revealed a missense mutation in the RDS/peripherin gene leading to a Tyr141Cys substitution. A phenotype and haplotype comparison between this and a separate family with the Tyr141Cys mutation suggested the presence of a common ancestral haplotype. CONCLUSIONS: The RDS mutation in codon 141 is associated with an unusual AMD-like late-onset maculopathy. An apparent selective bias was noted favoring the transmission of the mutant allele. These observations broaden the spectrum of phenotypes associated with RDS gene mutations.