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Rapid DNA library construction for functional genomic and metagenomic screening

A rapid protocol was developed for constructing plasmid libraries from small quantities of genomic/metagenomic DNA. The technique utilizes linker amplification with topoisomerase cloning and allows for inducible transcription in Escherichia coli. As proof of principle, several anti-Bacillus lysins were cloned from bacteriophage genomes and an aerolysin was cloned from a metagenomic sample.

No induction of thiopurine methyltransferase during thiopurine treatment in inflammatory bowel disease

The aim of this study was to follow, during standardized initiation of thiopurine treatment, thiopurine methyltransferase (TPMT) gene expression and enzyme activity and thiopurine metabolite concentrations, and to study the role of TPMT and ITPA 94C > A polymorphisms for the development of adverse drug reactions. Sixty patients with ulcerative colitis or Crohn's disease were included in this open

Solution structure of the SH3 domain from Bruton's tyrosine kinase

X-linked agammaglobulinemia (XLA) is a heritable immunodeficiency caused by mutations in the gene coding for Bruton's tyrosine kinase (Btk). Btk belongs to the Tec family of tyrosine kinases. Each member of the family contains five regions and mutations causing XLA have been isolated in all five regions. We have determined the solution structure of the Src homology 3 (SH3) domain of Btk using two-

Hunting reduces recruitment of primate-dispersed trees in Amazonian Peru

Hunting with firearms decimates primates of large and medium body size (> 2 kg) that disperse the seeds of large-seeded trees. In continuous, un-fragmented forests of southeastern Peru regularly hunted with firearms for 30-40 years, large primates are extirpated and medium-sized (medium) primates are reduced 61% compared with protected forests. At hunted sites seedlings and small juveniles (< 1 m

GRACILE syndrome, a lethal metabolic disorder with iron overload, is caused by a point mutation in BCS1L

GRACILE (growth retardation, aminoaciduria, cholestasis, iron overload, lactacidosis, and early death) syndrome is a recessively inherited lethal disease characterized by fetal growth retardation, lactic acidosis, aminoaciduria, cholestasis, and abnormalities in iron metabolism. We previously localized the causative gene to a 1.5-cM region on chromosome 2q33-37. In the present study, we report the

Sample preparation effects in matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry of partially depolymerised methyl cellulose

Methyl cellulose (MC) was partially depolymerised and the oligomers thus obtained were studied by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS). The depolymerisation was either enzymatic or acidic. Fractions of enzymatically depolymerised MC were collected from size-exclusion chromatography and subjected to a sample preparation investigation. Several MA

Generalised and Particularistic Thinking in Policy Analysis and Practice: The Case of Governance Reform in South Africa

This article is concerned with the relationship between generalised and particularistic knowledge in the context of policy-making and policy analysis. It argues that it is problematic to assume that a reform model will generate similar outcomes across a wide variety of contexts. It presents a conceptual framework, including the concepts of transaction domain and domain consensus, that enables cont

Defective glucose-stimulated insulin release in the diabetic Goto-Kakizaki (GK) rat coincides with reduced activity of the islet carbon monoxide signaling pathway

The Goto-Kakizaki (GK) rat displays a markedly reduced insulin response to glucose, a defect that is thought to be coupled to an impaired glucose signaling in the beta-cell. We have examined whether carbon monoxide (CO), derived from beta-cell heme oxygenase (HO), might be involved in the secretory dysfunction. Immunocytochemical labeling of constitutive HO (HO-2) showed no overt difference in flu

Transthyretin expression in the rat brain: effect of thyroid functional state and role in thyroxine transport

Rats were made hypo- or hyperthyroid to study the role of thyroid hormones on cerebral transthyretin (TTR) mRNA expression. TTR mRNA was detected by Northern blot in rat liver, choroid plexus and meninges but not in cultured astrocytes or cultured cerebral endothelial cells. No changes were found in the levels of TTR mRNA in liver, choroid plexus or meninges in hypo- or hyperthyroid rats compared

The consequences of the variance-mean rescaling effect on effective population size

The effective population size (N-e), and the ratio between N-e and census population size (N) are often used as measures of population viability. We show that using the harmonic mean of population sizes over time - a common proxy for N-e- has some important evolutionary consequences and implications for conservation management. This stems from the fact that there is no unambiguous relationship bet

Important variables for parents' postnatal sense of security: evaluating a new Swedish instrument (the PPSS instrument).

OBJECTIVE: to evaluate dimensions of both parents' postnatal sense of security the first week after childbirth, and to determine associations between the PPSS instrument and different sociodemographic and situational background variables. DESIGN: evaluative, cross-sectional design. PARTICIPANTS AND SETTINGS: 113 mothers and 99 fathers with children live born at term, from five hospitals in souther

Fungal populations in podzolic soil experimentally acidified to simulate acid-rain

Abstract in UndeterminedThe effect of experimental acidification on the soil microfungalcommunity was studied in the humus layer of a coniferous forest in northern Sweden. The study was made 4 years after the last application of sulfuric acid. Fungal species composition was altered by treatments of 100 and 150 kg sulfuric acid ha-1 each year for 6 years, yet no differences were found between the c

Motivation to donate or not donate surplus embryos for stem-cell research: literature review

Objective To review publications regarding possible donation of surplus embryos for medical research. Design Review of 67 scientific publications in this field up through January 2007. Originality Summary of factors influencing prospective donors' motivation to donate or not donate their surplus embryos for medical research. Result(s) Presentation of the findings of studies regarding the need

An origin for short gamma-ray bursts unassociated with current star formation

Two short (< 2 s) gamma-ray bursts (GRBs) have recently been localized(1-4) and fading afterglow counterparts detected(2-4). The combination of these two results left unclear the nature of the host galaxies of the bursts, because one was a star-forming dwarf, while the other was probably an elliptical galaxy. Here we report the X-ray localization of a short burst (GRB 050724) with unusual gamma-ra

Influence of initial C/N ratio on chemical and microbial composition during long term composting of straw

Shredded straw of Miscanthus was composted in 800-L boxes with different amounts of pig slurry added as nitrogen source. The impact of the different initial CIN ratios (11, 35, 47, 50, and 54) on the composting process and the end product was evaluated by examining chemical and microbiological parameters during 12 months of composting. Low initial C/N ratios caused a fast degradation of fibers dur