Display of functional alphabeta single-chain T-cell receptor molecules on the surface of bacteriophage

The ability to display functional T-cell receptors (TCR) on the surface of bacteriophage could have numerous applications. For instance, TCR phage-display could be used to develop new strategies for isolating TCRs with unique specificity or it could be used to carry out mutagenesis studies on TCR molecules for analyzing their structure-function. We initially selected a TCR from the murine T-cell hybridoma, DO11.10, as our model system, and genetically engineered a three domain single-chain TCR (scTCR) linked to the gene p8 protein of the Escherichia coli bacteriophage fd. Immunoblotting studies revealed that (1) E. coli produced a soluble scTCR/p8 fusion protein and (2) the fusion protein was packaged by the phage. Cellular competition assays were performed to evaluate the functionality of the TCR and showed the DO11.10 TCR-bearing phage could significantly inhibit stimulation of DO11.10 T hybridoma cells by competing for binding to immobilized MHC/peptide IA(d)/OVA(323-339). Flow cytometric analysis was carried out to evaluate direct binding of DO11.10 TCR-bearing phage onto the surface of cells displaying either IAd containing irrelevant peptide or OVA peptide. The results revealed binding of DO11.10 TCR-bearing phage only on cells expressing IA(d) loaded with OVA peptide showing TCR fine specificity for peptide. To illustrate the generality of TCR phage-display, we also cloned and displayed on phage a second TCR which recognizes a peptide fragment from human tumor suppressor protein p53 restricted by HLA-A2. These findings demonstrate functional TCR can be displayed on bacteriophage potentially leading to the development of novel applications involving TCR phage-display.     

Double blind, randomised, placebo controlled study of oral vancomycin in prevention of necrotising enterocolitis in preterm, very low birthweight infants

AIMS: To evaluate the effectiveness of oral vancomycin in the prophylaxis of necrotising enterocolitis in preterm, very low birthweight infants. METHODS: A prospective, double blind, randomised, placebo controlled study in a tertiary referral centre of a university teaching hospital was conducted on 140 very low birthweight infants consecutively admitted to the neonatal unit. The babies were randomly allocated to receive oral vancomycin (15 mg/kg every 8 hours for 7 days) or an equivalent volume of placebo solution. Prophylaxis was started 24 hours before the start of oral feeds. All suspected cases of necrotising enterocolitis were investigated with a full sepsis screen and serial abdominal radiographs. Necrotising enterocolitis was diagnosed and staged according to modified Bell's criteria. RESULTS: Nine of 71 infants receiving oral vancomycin and 19 of 69 infants receiving the placebo solution developed necrotising enterocolitis (p = 0.035). Infants with necrotising enterocolitis were associated with a significant increase in mortality (p = 0.026) and longer duration of hospital stay (p = 0.002). CONCLUSIONS: Prophylactic oral vancomycin conferred protection against necrotising enterocolitis in preterm, very low birthweight infants and was associated with a 50% reduction in the incidence. However, widespread implementation of this preventive measure is not recommended, as it would only be effective in necrotising enterocolitis caused by Gram positive organisms and could increase the danger of the emergence of vancomycin resistant or dependent organisms. Its use should be restricted to a high prevalence nursery for a short and well defined period in a selected group of high risk patients.     

Effect of mild acid treatment on the survival, enteropathogenicity, and protein production in vibrio parahaemolyticus

Vibrio parahaemolyticus is an important food-borne enteropathogen that encounters various adverse conditions in its native environment or during infection. Effects of mild acid treatment on survival under stress conditions, enteropathogenicity, and protein production in this pathogen were investigated. Logarithmically grown cells, at pH 7.5 shifted to pH 5.0 for 30 min, were more resistant to subsequent acid challenge at pH 4.4. A two-phase adaptive procedure (pH 5.8 for 30 min; pH 5.0 for 30 min) was better than a single-phase procedure for enhancing the acid tolerance of this pathogen. The acid-adapted cells were cross-protected against the challenges of low salinity and thermal inactivation. One-dimensional polyacrylamide gel electrophoresis revealed that proteins with molecular masses of 6.4, 9.0, 13.6, 16.3, 18.9, 22.9, 24.4, 28.3, 33. 9, 36.9, 41.2, 47.6, 58.1, 65.6, 80.5, 88.2, and 96.9 kDa were induced or significantly enhanced, while proteins of 25.3, 30.1, 30. 7, and 91.7 kDa were significantly inhibited. Two-dimensional polyacrylamide gel electrophoresis revealed that 20 species of proteins were induced or significantly enhanced, while 26 species were inhibited. In assays conducted using the suckling mouse model, enteropathogenicity of the acid-adapted cells was significantly enhanced in terms of intestine/body weight ratio and in vivo recovery of infected cells.     

In search of a dose-response relationship with radiotherapy in the management of recurrent rectal carcinoma in the pelvis: a systematic review

BACKGROUND: Research on the subject of insight has been hampered by difficulties in definition and reliable measurement. METHODS: We compared several rating scales to measure insight on a group of 33 psychotic patients as well as assessing patients' psychopathology, clinical characteristics and cognitive functioning. RESULTS: Most currently used scales showed a high degree of inter-correlation. Measures of insight related strongly to the presence of delusions; grandiosity (inversely), and depression (positively). Higher insight scores correlated with indices of treatment compliance and inversely with substance abuse. Measures of pre-morbid IQ and impaired executive functioning, including the Wisconsin Card Sorting Test were not associated with poor insight. CONCLUSIONS: The study highlights aspects of psychopathology and clinical variables particularly related to insight and supports the continued use of standardized scales in further research in this area.     

Effects of nitric oxide annealing of thermally grown silicondioxide characteristics

The effects of nitric oxide (NO) annealing on conventional thermal oxides are reported in this letter. The oxide thickness increase, resulting from NO annealing, is found to be only a few angstroms (<0.5 nm) and independent on the initial oxide thickness. Furthermore, both the electrical and physical characteristics are improved. This technique is expected to achieve sub-5 nm high quality ultrathin dielectric films for the applications in EEPROM's and ULSI     

Locality-preserving hash functions for general purpose parallel computation

Consider the problem of efficiently simulating the shared-memory parallel random access machine (PRAM) model on massively parallel architectures with physically distributed memory. To prevent network congestion and memory bank contention, it may be advantageous to hash the shared memory address space. The decision on whether or not to use hashing depends on (1) the communication latency in the network and (2) the locality of memory accesses in the algorithm.We relate this decision directly to algorithmic issues by studying the complexity of hashing in the Block PRAM model of Aggarwal, Chandra, and Snir, a shared-memory model of parallel computation which accounts for communication locality. For this model, we exhibit a universal family of hash functions having optimal locality. The complexity of applying these hash functions to the shared address space of the Block PRAM (i.e., by permuting data elements) is asymptotically equivalent to the complexity of performing a square matrix transpose, and this result is best possible for all pairwise independent universal hash families. These complexity bounds provide theoretical evidence that hashing and randomized routing need not destroy communication locality, addressing an open question of Valiant.This work was started when the author was a student at Oxford University, supported by a National Science Foundation Graduate Fellowship and a Rhodes Scholarship. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author and do not necessarily reflect the views of the National Science Foundation or the Rhodes Trust.     

A method for exploring the effects of attrition in randomized experiments with dichotomous outcomes.

Attrition from conditions in randomized experiments is common. Yet it is difficult to assess the possible effects of attrition because the outcome status of the dropouts is usually unknown. This article develops methods to assess those effects in studies with dichotomous outcomes, illustrating the methods with randomized experiments in drug abuse treatment, smoking cessation treatment, and alcoholism treatment. The methods include computing the lowest and highest possible effect sizes that could have been observed, enumerating the percent of possible study outcomes below a given threshold, estimating the probability that an outcome beyond any given threshold would be observed if all participants were measured, and constructing attrition analysis plots showing the effects of attrition under varied assumptions. For the kind of study to which they apply, these methods should replace the treatment of missing participants as failures in an "intent-to-treat" analysis. A user-friendly personal computer program is available to implement all of these analyses. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Analysis of a human cDNA containing a tissue-specific alternatively spliced LIM domain

A unique clone, isolated from a human pancreatic cDNA library, was sequenced and characterized. Northern blot analysis showed that the gene is active in a number of fetal and adult tissues, and immunoblots showed expression in nuclear and cytosolic cell fractions. The gene corresponding to the clone was localized to chromosome 13 by human/rodent somatic cell hybrid panels. The largest open reading frame contains a LIM domain, and the deduced peptide from the open reading frame appears to have the characteristics of a LIM-only protein, designated LMO7. RT-PCR and genomic sequence analyses indicate that expression of this gene product is subject to tissue-specific modulation by elimination of the LIM domain by alternative splicing in neural tissues.     

Spatially resolved X-ray diffraction phase mapping and α → β → α transformation kinetics in the heat-affected zone of commercially pure titanium arc welds

Spatially resolved X-ray diffraction (SRXRD) is used to map the α → β → α phase transformation in the heat-affected zone (HAZ) of commercially pure titanium gas tungsten arc welds. In situ SRXRD experiments were conducted using a 180-μm-diameter X-ray beam at the Stanford Synchrotron Radiation Laboratory (SSRL) (Stanford, CA) to probe the phases present in the HAZ of a 1.9 kW weld moving at 1.1 mm/s. Results of sequential linear X-ray diffraction scans made perpendicular to the weld direction were combined to construct a phase transformation map around the liquid weld pool. This map identifies six HAZ microstructural regions between the liquid weld pool and the base metal: (1) α-Ti that is undergoing annealing and recrystallization; (2) completely recrystallized α-Ti; (3) partially transformed α-Ti, where α-Ti and β-Ti coexist; (4) single-phase β-Ti; (5) back-transformed α-Ti; and (6) recrystallized α-Ti plus back-transformed α-Ti. Although the microstructure consisted predominantly of α-Ti, both prior to and after the weld, the crystallographically textured starting material was altered during welding to produce different α-Ti textures within the resulting HAZ. Based on the travel speed of the weld, the α → β transformation was measured to take 1.83 seconds during heating, while the β → α transformation was measured to take 0.91 seconds during cooling. The α → β transformation was characterized to be dominated by long-range diffusional growth on the leading (heating) side of the weld, while the β → α transformation was characterized to be predominantly massive on the trailing (cooling) side of the weld, with a massive growth rate on the order of 100 μm/s.