Aberrant TAL1 activation is mediated by an interchromosomal interaction in human T-cell acute lymphoblastic leukemia.

Long-range chromatin interactions control metazoan gene transcription. However, the involvement of intra- and interchromosomal interactions in development and oncogenesis remains unclear. TAL1/SCL is a critical transcription factor required for the development of all hematopoietic lineages; yet, aberrant TAL1 transcription often occurs in T-cell acute lymphoblastic leukemia (T-ALL). Here, we report that oncogenic TAL1 expression is regulated by different intra- and interchromosomal loops in normal hematopoietic and leukemic cells, respectively. These intra- and interchromosomal loops alter the cell-type-specific enhancers that interact with the TAL1 promoter. We show that human SET1 (hSET1)-mediated H3K4 methylations promote a long-range chromatin loop, which brings the +51 enhancer in close proximity to TAL1 promoter 1 in erythroid cells. The CCCTC-binding factor (CTCF) facilitates this long-range enhancer/promoter interaction of the TAL1 locus in erythroid cells while blocking the same enhancer/promoter interaction of the TAL1 locus in human T-cell leukemia. In human T-ALL, a T-cell-specific transcription factor c-Maf-mediated interchromosomal interaction brings the TAL1 promoter into close proximity with a T-cell-specific regulatory element located on chromosome 16, activating aberrant TAL1 oncogene expression. Thus, our study reveals a novel molecular mechanism involving changes in three-dimensional chromatin interactions that activate the TAL1 oncogene in human T-cell leukemia.

A survey of infection prevention and control resources in acute care facilities across British Columbia.

BACKGROUND: To determine the gaps in infection prevention and control (IPAC) resources and the disparities between rural and urban areas, the Provincial Infection Control Network surveyed the current resources in British Columbia (BC). METHODS: Acute care facilities (ACF) in six health authorities (HA) were surveyed for IPAC staff; distribution of work; infection prevention and control professional (ICP) to bed ratios; and teaching activities. HAs were designated as either urban or rural. RESULTS: Responses represented 54 (68%) of the ACF in BC. Rural HAs showed a significantly higher number of inexperienced ICPs (68% vs. 17%; p < 0.001). Only 22 (60%) of eligible ICPs were Certification Board of Infection Control certified. Five out of six HAs (83%) reported having an IPAC physician. Acute care ICP to bed ratios ranged from one per 67 to one per 175 and combined acute and long-term care ICP to bed ratios ranged from one per 270 beds to one per 525 beds. The number of ICPs who reported working overtime on a consistent basis ranged from 20 to 100%. CONCLUSIONS: ACFs surveyed did not meet the recommended standards for staffing and IPAC resources in order to function as an effective program. Surveys of infection control resources are valuable tools to identify needs and assist in acquiring the resources to fill the identified gaps within a health authority.

Fifty-four new gene-based canine microsatellite markers.

Fifty-four new markers were developed to fill in gaps in the current map of canine microsatellites and to complement existing markers that may not be sufficiently informative in highly inbred canine pedigrees. Canine genes contained on the radiation hybrid map were used to obtain the sequence of the human homolog. A BLAST search versus the canine whole genome shotgun (wgs) sequence resource was used to obtain the sequence of the canine genomic contigs containing the homolog of the corresponding human gene. Canine sequences that contained microsatellites and mapped back to the correct location in the human genome were used to design primers for amplification of the microsatellites from canine genomic DNA. Heterozygosities of the markers were tested by genotyping grandparental DNAs obtained from the Nestle Purina Reference family DNA distribution center plus DNAs from unrelated Bouviers and Irish wolfhounds. Canine map positions of markers on the July 2004 freeze of the canine genome assembly were determined by in silico PCR or BLAST.

UK budgetary systems and new health-care technologies.

OBJECTIVES: This article outlines the budgetary setting within the UK health-care system. METHODS: It is argued that while prospective budgets can give rise to efficient resource allocation outcomes, this relies on the budget being set at an appropriate level and the accompanying incentive structures being efficient. The organizational structures and the interrelationships are critical. The recent history of UK National Health Service reforms and expenditure is outlined. It is suggested that until recently, although the budget system has the potential to promote efficiency, the aggregate budget allocated to the NHS has probably been too low given public expectations, technology advances, and preferences for health care. RESULTS: The aggregate budget is due to rise considerably over the next 5 years. While some incentive and regulatory provisions will move the budget toward an efficient allocation many microlevel incentive issues remain. CONCLUSION: Whether efficient patterns of health-care allocation emerge remains open to debate, however, because the existing incentive mechanisms are not optimal.

Novel lamin A/C mutations in two families with dilated cardiomyopathy and conduction system disease.

BACKGROUND: The LMNA gene, one of 6 autosomal disease genes implicated in familial dilated cardiomyopathy, encodes lamins A and C, alternatively spliced nuclear envelope proteins. Mutations in lamin A/C cause 4 diseases: Emery-Dreifuss muscular dystrophy, limb girdle muscular dystrophy type 1B, Dunnigan-type familial partial lipodystrophy, and dilated cardiomyopathy. METHODS AND RESULTS: Two 4-generation white families with autosomal dominant familial dilated cardiomyopathy and conduction system disease were found to have novel mutations in the rod segment of lamin A/C. In family A a missense mutation (nucleotide G607A, amino acid E203K) was identified in 14 adult subjects; disease was manifest as progressive conduction disease in the fourth and fifth decades. Death was caused by heart failure. In family B a nonsense mutation (nucleotide C673T, amino acid R225X) was identified in 10 adult subjects; disease was also manifest as progressive conduction disease but with earlier onset (third and fourth decades), ventricular dysrhythmias, left ventricular enlargement, and systolic dysfunction. Death was caused by heart failure and sudden cardiac death. Skeletal muscle disease was not observed in either family. CONCLUSIONS: Novel rod segment mutations in lamin A/C cause variable conduction system disease and dilated cardiomyopathy without skeletal myopathy.

Correlation between histone lysine methylation and developmental changes at the chicken beta-globin locus.

Methylation of histones at specific residues plays an important role in transcriptional regulation. Chromatin immunoprecipitation of dimethylated lysine 9 on histone H3 across 53 kilobases of the chicken beta-globin locus during erythropoiesis shows an almost complete anticorrelation between regions of elevated lysine 9 methylation and acetylation. Lysine 9 is methylated most over constitutive condensed chromatin and developmentally inactive globin genes. In contrast, lysine 4 methylation of histone H3 correlates with H3 acetylation. These results lead us to propose a mechanism by which the insulator in the beta-globin locus can protect the globin genes from being silenced by adjacent condensed chromatin.

Prospective matching of alcoholic clients to cognitive-behavioral or interactional group therapy.

OBJECTIVE: In earlier work, client sociopathy and global psychopathology were effective variables for treatment matching: clients low on both sociopathy and severity of psychopathology were likely to benefit from interactional group therapy, whereas those scoring high on either of these dimensions benefited more from a coping skills intervention. The present study assessed whether outcomes improve further when clients are assigned to group treatments prospectively based on a matching strategy derived from the previous findings. METHOD: All participants (N = 250, 66% men) met criteria for alcohol dependence or abuse. About half were prospectively assigned to either cognitive-behavioral (CB) coping skills training or interactional therapy, those with higher levels of psychiatric severity or sociopathy were given CB and those who were low on both dimensions were given interactional therapy. The other half were randomly assigned to those treatments, replicating the procedure of the earlier study. Outcome data were collected at the conclusion of treatment and at 3-month intervals for 1 year following. RESULTS: Prospective matching of clients to treatment did not produce superior drinking outcomes compared to random treatment assignment. Randomly assigned clients were more likely to be abstinent at the end of treatment, but this effect disappeared at later follow-ups. Prospectively matched clients had fewer negative consequences of drinking than did those assigned randomly (unmatched). Neither sociopathy nor psychiatric severity was particularly effective for matching. CONCLUSIONS: The matching effects from our previous study were not replicated. Nevertheless, prospective matching did reduce the negative consequences of drinking, consistent with our previous results.

Transitions in histone acetylation reveal boundaries of three separately regulated neighboring loci.

We have studied developmentally regulated patterns of histone acetylation at high resolution across approximately 54 kb of DNA containing three independently regulated but neighboring genetic loci. These include a folate receptor gene, a 16 kb condensed chromatin region, the chicken beta-globin domain and an adjacent olfactory receptor gene. Within these regions the relative levels of acetylation appear to fall into three classes. The condensed chromatin region maintains the lowest acetylation at every developmental stage. Genes that are inactive show similarly low levels, but activation results in a dramatic increase in acetylation. The highest levels of acetylation are seen at regulatory sites upstream of the genes. These patterns imply the action of more than one class of acetylation. Notably, there is a very strong constitutive focus of hyperacetylation at the 5' insulator element separating the globin locus from the folate receptor region, which suggests that this insulator element may harbor a high concentration of histone acetylases.

Characterization of (CA)n microsatellite repeats from large-insert clones.

The most laborious part of developing (CA)n microsatellite repeats as genetic markers is constructing DNA clones to permit determination of sequences flanking the microsatellites. When cosmids or large-insert phage clones are used as primary sources of (CA)n repeat markers, they have traditionally been subcloned into plasmid vectors such as pUC18 or M13 mp 18/19 cloning vectors to obtain fragments of suitable size for DNA sequencing. This unit presents an alternative approach whereby a set of degenerate sequencing primers that anneal directly to (CA)n microsatellites can be used to determine sequences that are inaccessible with vector-derived primers. Because the primers anneal to the repeat and not to the vector, they can be used with subclones containing inserts of several kilobases and should, in theory, always give sequence in the regions directly flanking the repeat. Degeneracy at the 3 end of each of these primers prevents elongation of primers that have annealed out-of-register. The most laborious part of developing (CA)n microsatellite repeats as genetic markers is constructing DNA clones to permit.

Clinical and psychological correlates of lumbar motion abnormalities in low back disorders.

BACKGROUND CONTEXT: Low back pain (LBP) and low back disorders (LBDs) identify a complex constellation of conditions that frustrate both diagnosis and therapy. Dynamic quantitative assessment and questionnaire instruments directed toward psychosocial and situational variables provide potentially powerful tools for determining functional pathology and potentially outcome. PURPOSE: Our goal was to independently assess clinical correlates of a trunk motion measurement device, the lumbar motion monitor (LMM). The reliability of the LMM as a clinical test was assessed by comparison with an independent medical examination and biobehavioral questionnaires. STUDY DESIGN/SETTING: There were three study components. A multispecialty physician panel that administered a structured physical examination contributed to a clinical correlation case series study. Standardized outcomes and risk identification questionnaires were administered to the case population. Finally, the LMM was administered in a customary fashion to the same population. PATIENT SAMPLE: Nineteen subjects were recruited on the basis of criteria that included symptoms of chronic recurrent low back pain. This was an employed and active, although impaired, population. Eighteen of the subjects were currently employed with limited lost work time, but chronic and recurrent pain was a common feature. OUTCOME MEASURES: Questionnaire outcome measures were both characterologic and situation based. In addition to providing diagnoses, the physician panel was also asked to offer certain qualitative assessments, such as rehabilitative potential and functional level pertinent to activities of daily living. The impact of LMM measures on physician decision making was also assessed. Trunk angular measurements were used to assess function of patients with chronic low back disorders. METHODS: Kinematic performance on the LMM was expressed as three probability scores. These were the likelihood of abnormality, the "sincerity of effort" (exacerbation or aggravation of impairment), and the likelihood of structural anatomic disease. These variables were examined against established self-report measures of pain and disability. RESULTS: The LMM and physician panels were in agreement on the presence or absence of abnormality. LMM findings tended to be more consistent with clinical history than the clinical examination. The LMM results were also generally consistent with the self-reported measures of pain and disability: a high likelihood of structural disease was associated with depression, somatization, poor health perception and diminished vitality. CONCLUSIONS: The LMM appears to be a useful assessment tool for gauging the presence of LBP and LBD. It was accurate in detecting abnormality when abnormality was determined by clinical history and physician diagnosis. The LMM's differentiation of mechanical low back disease (nonanatomically specific disorders) from structurally specific low back disease was not consistent with a parallel clinical differentiation. Larger trials in a prospective format and studies on a chronically disabled population seem warranted. In an impaired but less disabled population, elevated pain and somatization did not appear to weaken the effort during testing.

Numerical model and experimental validation of microcarrier motion in a rotating bioreactor.

The equations of motion for microcarriers in a rotating bioreactor have been formulated and trajectories obtained using numerical techniques. An imaging system was built to validate the results by direct observation of microcarrier trajectories in the rotating frame of reference. The microcarrier motion observed by this imaging system was in excellent agreement with the numerical predictions of that motion. In the rotating frame of reference, microcarriers with density greater than the surrounding fluid medium followed a circular motion relative to the culture medium combined with a persistent migration and eventual collision with the outer wall of the reactor. However, for microcarrier density less the fluid medium, their circular motion migrated toward the central region of the reactor. When multiple microcarrier beads that are lighter than water are inserted into the reactor, the centrally directed migration results in the formation of clusters that are stabilized by tissue bridges formed by osteoblasts seeded onto the microcarriers. This system offers unique opportunities to monitor tissue synthesis on microcarriers using real-time optical techniques and to optimize the bioreactor operating conditions for exploiting this technology to study early bone tissue synthesis in vitro.

Reactivity to alcohol-related stimuli in the laboratory and in the field: predictors of craving in treated alcoholics.

AIMS: (1) To search for predictors of alcohol craving in treated alcoholics; (2) to evaluate the relationship between craving and drinking immediately after treatment. DESIGN: Alcoholic patients in treatment underwent cue-reactivity trails in the laboratory and then recorded craving in the field using hand-held computers. Laboratory craving was correlated with craving in the field, and moods and situations recorded in the field were correlated with contemporaneous craving ratings using a multi-level correlational design. SETTING: A VA Medical Center substance abuse treatment program provided the treatment and laboratory settings. The patients' home environment was the field setting. PARTICIPANTS: Male alcohol-dependent veterans (N = 26) treated in a VA inpatient or intensive outpatient program. INTERVENTION: Participants underwent two cue-reactivity laboratory sessions prior to discharge to measure craving. Following discharge, participants recorded drinking and cravings eight times per day for 21 consecutive days. MEASUREMENTS: Craving ratings in the laboratory and multiple recordings per day of surroundings, craving and mood state in the field. FINDINGS: Desire to drink in the laboratory accounted for 8-10% of the variance in later drinking and urges to drink recorded in the field--a modest correlation. Frequency of positive urges in the field was significantly correlated with drinking frequency. Those who reported urges in the field had greater alcohol dependence and higher trait anger and anxiety scores than non-reporters. CONCLUSIONS: Craving is related to drinking immediately following treatment, and is most likely in those who have more severe dependence and greater mood disturbance. These individuals may benefit most from interventions for coping with cravings after treatment.

Paroxysmal kinesigenic dyskinesia and infantile convulsions: clinical and linkage studies.

OBJECTIVE: To clinically characterize affected individuals in families with paroxysmal kinesigenic dyskinesia (PKD), examine the association with infantile convulsions, and confirm linkage to a pericentromeric chromosome 16 locus. BACKGROUND: PKD is characterized by frequent, recurrent attacks of involuntary movement or posturing in response to sudden movement, stress, or excitement. Recently, an autosomal dominant PKD locus on chromosome 16 was identified. METHODS: The authors studied 11 previously unreported families of diverse ethnic background with PKD with or without infantile convulsions and performed linkage analysis with markers spanning the chromosome 16 locus. Detailed clinical questionnaires and interviews were conducted with affected and unaffected family members. RESULTS: Clinical characterization and sampling of 95 individuals in 11 families revealed 44 individuals with paroxysmal dyskinesia, infantile convulsions, or both. Infantile convulsions were surprisingly common, occurring in 9 of 11 families. In only two individuals did generalized seizures occur in later childhood or adulthood. The authors defined a 26-cM region using linkage data in 11 families (maximum lod score 6.63 at theta = 0). Affected individuals in one family showed no evidence for a shared haplotype in this region, implying locus heterogeneity. CONCLUSIONS: Identification and characterization of the PKD/infantile convulsions gene will provide new insight into the pathophysiology of this disorder, which spans the phenotypic spectrum between epilepsy and movement disorder.

Autosomal-dominant congenital cataract associated with a deletion mutation in the human beaded filament protein gene BFSP2.

Congenital cataracts are a common major abnormality of the eye that frequently cause blindness in infants. At least one-third of all cases are familial; autosomal-dominant congenital cataract appears to be the most-common familial form in the Western world. Elsewhere, in family ADCC-3, we mapped an autosomal-dominant cataract gene to chromosome 3q21-q22, near the gene that encodes a lens-specific beaded filament protein gene, BFSP2. By sequencing the coding regions of BFSP2, we found that a deletion mutation, DeltaE233, is associated with cataracts in this family. This is the first report of an inherited cataract that is caused by a mutation in a cytoskeletal protein.

A new locus for autosomal dominant congenital cataracts maps to chromosome 3.

PURPOSE: To map a gene for cataracts in a family with congenital nuclear and sutural cataracts and to examine candidate genes in the linked region. METHODS: A large family with autosomal dominant congenital nuclear and sutural cataracts was identified and characterized. A genome-wide screen was conducted with a set of markers spaced at 10- to 15-cM intervals, and linkage was assessed using standard LOD score analysis. RESULT: Fifteen (15) affected individuals were identified. This form of congenital cataracts maps to a 12-cM region on chromosome 3q21.2-q22.3 between markers D3S3674 and D3S3612, with a maximum multipoint LOD score of 6.94 at D3S1273. The crystallin gene, CRYGS, was excluded as a candidate gene for this locus. CONCLUSIONS: There are now more than 12 different genetic loci that cause congenital cataracts. The most recent locus to be identified is on chromosome 3q21.2-q22.3, in a family with congenital nuclear and sutural cataracts.

Gene structure and chromosome mapping of the human small-conductance calcium-activated potassium channel SK1 gene (KCNN1).

Small-conductance, calcium-activated potassium channels contribute to the afterhyperpolarization in central neurons and other cell types. Because these channels regulate neuronal excitability, defects in their genes could cause excitability disorders. The human cDNA encoding one such channel, SK1 (KCNN1), was recently cloned. Here we describe the gene structure of KCNN1 and its localization by radiation hybrid mapping to chromosome 19p13.1.

A novel nonsense mutation in CACNA1A causes episodic ataxia and hemiplegia.

OBJECTIVE: To identify the disease-causing mutation and to characterize penetrance and phenotypic variability in a large pedigree with episodic ataxia type 2 (EA-2) previously linked to chromosome 19. BACKGROUND: Mutations in the CACNA1A gene on chromosome 19 encoding a calcium channel subunit cause EA-2, which is characterized by recurrent attacks of imbalance with interictal eye movement abnormalities. METHODS: The authors used single-strand conformation polymorphism (SSCP) analysis to screen for point mutations, and direct sequencing to identify mutations in CACNA1A. Allele-specific oligonucleotides were designed to detect the presence of the diseased allele in members of their pedigree as well as in normal control subjects. RESULTS: Reassessment of members of the pedigree revealed two notable clinical features. Diffuse weakness during attacks of ataxia was a prominent complaint. Two affected individuals had had episodic hemiplegia, one with typical migraine headaches. SSCP analysis revealed aberrant bands in exon 29 in affected members but not in normal control subjects. Direct sequencing of exon 29 identified a C-to-T change at position 4914 of the coding sequence of CACNA1A, predicting an early stop code at codon 1547. Two asymptomatic mutation carriers demonstrated the incomplete penetrance of this mutation. CONCLUSIONS: A nonsense mutation in CACNA1A causes episodic ataxia and complaint of weakness, and may be associated with hemiplegia.