Clinically meaningful blood pressure reductions with low intensity isometric handgrip exercise. A randomized trial.

There exists no examination of what is the minimum anti-hypertensive threshold intensity for isometric exercise training. Twenty two normotensive participants were randomly assigned to training intensities at either 5 % or 10 % of their maximal contraction. Twenty participants completed the study. Clinical meaningful, but not statistically significant, reductions in systolic blood pressure were observed in both 5 % and 10 % groups -4.04 mm Hg (95 % CI -8.67 to +0.59, p=0.08) and -5.62 mm Hg (95 % CI -11.5 to +0.29, p=0.06) respectively after 6 weeks training. No diastolic blood pressure reductions were observed in either 5 % -0.97 mm Hg (95 % CI -2.56 to +0.62, p=0.20) or 10 % MVC +1.8 mm Hg (95 % CI -1.29 to +4.89, p=0.22) groups respectively after training. In those unable to complete isometric exercise at the traditional 30 % intensity, our results suggest there is no difference between 5 and 10 % groups and based on the principle of regression to the mean, this could mean both interventions induce a similar placebo-effect.

Cost-effectiveness of venous thromboembolism prophylaxis in total hip and knee replacement surgery: the evolving application of health economic modelling over 20 years.

OBJECTIVES: In the last two decades, there has been considerable evolution of methods for cost-effectiveness modelling. Some of the first models were developed in the area of venous thromboembolism (VTE) prophylaxis. Hence, this area can serve as an important example to illustrate evolving standards. Our objectives are to document evolving methodology by describing VTE models, assess their critical strengths and weaknesses, and inform future advances for models in this therapeutic area. RESEARCH DESIGN AND METHODS: A systematic review of economic models of primary VTE prevention following hip and knee replacement surgery was undertaken. Electronic searches of PubMed, EMBASE, the Cochrane library, and grey literature were conducted (1985-2006). Reference lists of included articles and reviews were examined for relevant studies. RESULTS: Twenty-nine cost-effectiveness models were identified. Nineteen other cost-effectiveness analyses were excluded because they were not model-based; 16 were simple cost calculations and three were analyses of resource use data collected alongside clinical trials. The majority of models (24) were constructed as decision trees, frequently utilising previously published model structures, with some adaptation for new comparators, and/or addition of relevant events omitted by earlier models (e.g., bleeding due to prophylactic treatment). Later models have included Markov processes to model potential long-term consequences of VTE (recurrent VTE and post-thrombotic syndrome) over longer time horizons. Systematic identification of clinical evidence and more sophisticated analysis methods (e.g., Bayesian mixed-treatment comparisons and probabilistic sensitivity analyses) have recently been introduced. CONCLUSIONS: Model structures have evolved substantially in this highly studied therapeutic area, with improvements made to the model structure, the comprehensiveness of clinical evidence included, and the underlying calculation methodology.

Cerebral blood volume changes during closure by surgery of patent ductus arteriosus.

BACKGROUND: Surgical closure of a patent ductus arteriosus (PDA) in very low birth weight (VLBW) infants has been associated with impaired neurodevelopmental outcome. Surgical PDA closure may result in abrupt changes of cerebral haemodynamics. OBJECTIVE: To examine the cerebral blood volume changes occurring after surgical closure of PDA. DESIGN: Continuous cerebral near-infrared spectroscopy (NIRS) recording throughout PDA surgery. SETTING: Tertiary neonatal intensive care unit, with PDA surgery performed on the ward. PATIENTS: Ten VLBW infants, median birth weight 748 g (range 590-1070), gestational age 24 (23-27) weeks, chronological age 14 (12-22) days. INTERVENTION: Surgical closure of PDA. MAIN OUTCOME MEASURES: Changes in cerebral oxygenated haemoglobin, cerebral deoxygenated haemoglobin, and tissue oxygenation index (measured), changes in cerebral blood volume (CBV) and cerebral haemoglobin difference (calculated) as measured by NIRS. RESULTS: During the first 2 minutes after closure of the PDA, CBV increased significantly (mean (SD) 0.14 (0.12) ml/100 g tissue; p = 0.01) and returned to baseline within 2-5 minutes. Cerebral oxygenation did not change. CONCLUSIONS: There is a short-lasting increase in CBV immediately after surgical closure of PDA, but no change in cerebral oxygenation. These transient changes are unlikely to cause harm.

Treatment of therapy-resistant Alopecia areata with fumaric acid esters.

BACKGROUND: Alopecia areata is a cosmetically very disfiguring clinical picture and can be a great emotional burden to the patient, especially when persisting for a longer period of time. PATIENTS AND METHODS: 10 patients with an alopecia resistant to therapy were treated within the bounds of an open, non-placebo controlled pilot study with fumaric acid esters (FAE's, Fumaderm) for a period of six months and a maximum dose of 120 mg dimethylfumarate per day. The shortest space of time between persistent Alopecia areata and the start of the therapy with FAE was between six months and 17 years. RESULTS: Six patients took benefit from the six months therapy with FAE. In three of them very good results could be observed, presenting an almost entire remission, one patient showed a good success with a focal remission. With two patients a mediocre to moderate outcome was observed with growth of partly diffuse spread or very thin hair. Four patients took no benefit from the FAE therapy at all. CONCLUSIONS: FAE can be useful in the treatment of therapy-resistant Alopecia areata. This therapy approach should be validated in a multi-centre study.

Technetium reduction in sediments of a shallow aquifer exhibiting dissimilatory iron reduction potential.

Pertechnetate ion [Tc(VII)O(4) (-)] reduction rate was determined in core samples from a shallow sandy aquifer located on the US Atlantic Coastal Plain. The aquifer is generally low in dissolved O(2) (<1 mg L(-1)) and composed of weakly indurated late Pleistocene sediments differing markedly in physicochemical properties. Thermodynamic calculations, X-ray absorption spectroscopy and statistical analyses were used to establish the dominant reduction mechanisms, constraints on Tc solubility, and the oxidation state, and speciation of sediment reduction products. The extent of Tc(VII) reduction differed markedly between sediments (ranging from 0% to 100% after 10 days of equilibration), with low solubility Tc(IV) hydrous oxide the major solid phase reduction product. The dominant electron donor in the sediments proved to be (0.5 M HCl extractable) Fe(II). Sediment Fe(II)/Tc(VII) concentrations >4.3 were generally sufficient for complete reduction of Tc(VII) added [1-2.5 micromol (dry wt. sediment) g(-1)]. At these Fe(II) concentrations, the Tc (VII) reduction rate exceeded that observed previously for Fe(II)-mediated reduction on isolated solids of geologic or biogenic origin, suggesting that sediment Fe(II) was either more reactive and/or that electron shuttles played a role in sediment Tc(VII) reduction processes. In buried peats, Fe(II) in excess did not result in complete removal of Tc from solution, perhaps because organic complexation of Tc(IV) limited formation of the Tc(IV) hydrous oxide. In some sands exhibiting Fe(II)/Tc(VII) concentrations <1.1, there was presumptive evidence for direct enzymatic reduction of Tc(VII). Addition of organic electron donors (acetate, lactate) resulted in microbial reduction of (up to 35%) Fe(III) and corresponding increases in extractable Fe(II) in sands that exhibited lowest initial Tc(VII) reduction and highest hydraulic conductivities, suggesting that accelerated microbial reduction of Fe(III) could offer a viable means of attenuating mobile Tc(VII) in this type of sediment system.

Influence of salivary pellicle formation time on enamel demineralization--an in situ pilot study.

This study assessed the protective potential of salivary pellicles formed in situ over periods ranging from 2 to 24 h. Pellicles were produced on enamel slabs mounted on the palatal aspect of removable acrylic splints and exposed to the oral environment in three subjects for 2, 6, 12 and 24 h. Enamel specimens with and without pellicles were immersed in citric acid (1%) for 60 s, and the amount of dissolved calcium was measured by atomic absorption spectroscopy. In addition, specimens were processed for transmission electron microscopy (TEM). Mean values (standard deviations) for calcium release (mg/l related to the specimen's surface area of 5 x 5 mm(2)) were: 2-h pellicle 6.94 (1.55); 6-h pellicle 6.69 (2.05); 12-h pellicle 6.57 (2.31); 24-h pellicle 5.71 (2.46); enamel without pellicle 8.95 (1.66). There were no significant differences in calcium release that were dependent on pellicle formation time, but in comparison to enamel specimens without pellicle, significantly less (p <0.05) demineralization of the enamel was observed in pellicle-covered specimens. TEM showed that the pellicle was partly, but not completely dissolved following acid exposure. It is concluded that even a 2-h in-situ-formed pellicle layer protects the enamel surface to a certain extent against demineralization.

Spectroscopic studies of zinc(II)- and cobalt(II)-associated Escherichia coli formamidopyrimidine-DNA glycosylase: extended X-ray absorption fine structure evidence for a metal-binding domain.

Formamidopyrimidine-DNA glycosylase (Fpg) is a 30.2 kDa protein that plays an important role in the base excision repair of oxidatively damaged DNA in Escherichia coli. Sequence analysis and genetic evidence suggest that zinc is associated with a C4-type motif, C(244)-X(2)-C(247)-X(16)-C(264)-X(2)-C(267), located at the C-terminus of the protein. The zinc-associated motif has been shown to be essential for damaged DNA recognition. Extended X-ray absorption fine structure (EXAFS) spectra collected on the zinc-associated protein (ZnFpg) in the lyophilized state and in 10% frozen aqueous glycerol solution show directly that the metal is coordinated to the sulfur atom of four cysteine residues. The average Zn-S bond length is 2.33 +/- 0.01 and 2.34 +/- 0.01 A, respectively, in the lyophilized state and in 10% frozen aqueous glycerol solution. Fpg was also expressed in minimal medium supplemented with cobalt nitrate to yield a blue-colored protein that was primarily cobalt-associated (CoFpg). The profiles of the circular dichroism spectra for CoFpg and ZnFpg are identical, suggesting that the substitution of Co(2+) for Zn(2+) does not alter the structure of Fpg. A similar conclusion is reached upon the analysis of two-dimensional (15)N/(1)H HSQC spectra of uniformly (15)N-labeled samples of ZnFpg and CoFpg; the spectra are similar and display features characteristic of a structured protein. Biochemical assays with a 54 nt DNA oligomer containing 7, 8-dihydro-8-oxoguanine at a specific location show that CoFpg and ZnFpg are equally active at cleaving the DNA at the site of the oxidized guanine. EXAFS spectra of CoFpg indicate that the cobalt is coordinated to the sulfur atom of four cysteine residues with an average Co-S bond length of 2.28 +/- 0.01 and 2.29 +/- 0.01 A, respectively, in the lyophilized state and in 10% frozen aqueous glycerol solution. The structural similarity between CoFpg and ZnFpg suggests that it is biologically relevant to use the paramagnetic properties of Co(2+) as a structural probe.

Effect of electron donor and solution chemistry on products of dissimilatory reduction of technetium by Shewanella putrefaciens.

To help provide a fundamental basis for use of microbial dissimilatory reduction processes in separating or immobilizing (99)Tc in waste or groundwaters, the effects of electron donor and the presence of the bicarbonate ion on the rate and extent of pertechnetate ion [Tc(VII)O(4)(-)] enzymatic reduction by the subsurface metal-reducing bacterium Shewanella putrefaciens CN32 were determined, and the forms of aqueous and solid-phase reduction products were evaluated through a combination of high-resolution transmission electron microscopy, X-ray absorption spectroscopy, and thermodynamic calculations. When H(2) served as the electron donor, dissolved Tc(VII) was rapidly reduced to amorphous Tc(IV) hydrous oxide, which was largely associated with the cell in unbuffered 0. 85% NaCl and with extracellular particulates (0.2 to 0.001 microm) in bicarbonate buffer. Cell-associated Tc was present principally in the periplasm and outside the outer membrane. The reduction rate was much lower when lactate was the electron donor, with extracellular Tc(IV) hydrous oxide the dominant solid-phase reduction product, but in bicarbonate systems much less Tc(IV) was associated directly with the cell and solid-phase Tc(IV) carbonate may have been present. In the presence of carbonate, soluble (<0.001 microm) electronegative, Tc(IV) carbonate complexes were also formed that exceeded Tc(VII)O(4)(-) in electrophoretic mobility. Thermodynamic calculations indicate that the dominant reduced Tc species identified in the experiments would be stable over a range of E(h) and pH conditions typical of natural waters. Thus, carbonate complexes may represent an important pathway for Tc transport in anaerobic subsurface environments, where it has generally been assumed that Tc mobility is controlled by low-solubility Tc(IV) hydrous oxide and adsorptive, aqueous Tc(IV) hydrolysis products.

Cadmium mutagenicity and human nucleotide excision repair protein XPA: CD, EXAFS and (1)H/(15)N-NMR spectroscopic studies on the zinc(II)- and cadmium(II)-associated minimal DNA-binding domain (M98-F219).

Human XPA is a 31 kDa protein involved in nucleotide excision repair (NER), a ubiquitous, multi-enzyme pathway responsible for processing multiple types of DNA damage in the eukaryotic genome. A zinc-associated, C4-type motif (C105-X(2)-C108-X(17)-C126-X(2)-C129) located in the minimal DNA-binding region (M98-F219) of XPA (XPA-MBD) is essential for damaged DNA recognition. Cadmium is a known carcinogen and can displace the zinc in many metal-binding proteins. It has been suggested that the carcinogenic properties of cadmium may result from structural changes effected in XPA when Cd(2+) is substituted for Zn(2+) in the metal-binding site. The solution structure of XPA-MBD containing zinc(II) has recently been determined [Buchko et al., (1998) Nucleic Acids Res., 26, 2779-2788; Buchko et al., (1999) Biochemistry, 38, 15116-15128]. To assess the effects of cadmium(II) substitution on the structure of XPA-MBD, XPA-MBD was expressed in minimal medium supplemented with cadmium acetate to yield a protein that was almost exclusively (>95%) associated with cadmium(II) (CdXPA-MBD). Extended X-ray absorption fine structure spectra collected on ZnXPA-MBD and CdXPA-MBD in frozen (77 K) 15% aqueous glycerol solution show that the metal is coordinated to the sulfur atoms of four cysteine residues with an average metal-sulfur bond length of 2.34 +/- 0.01 and 2.54 +/- 0.01 A, respectively. Comparison of the circular dichroism, two-dimensional (1)H,(15)N-HSQC, and three-dimensional (15)N-edited HSQC-NOESY spectra of ZnXPA-MBD and CdXPA-MBD show that there are no structural differences between the two proteins. The absence of major structural changes upon substituting cadmium(II) for zinc(II) in XPA suggests that cadmium-induced mutagenesis is probably not due to structural perturbations to the zinc-binding core of XPA.

Extended X-ray absorption fine structure evidence for a single metal binding domain in Xenopus laevis nucleotide excision repair protein XPA.

Nucleotide excision repair (NER) is an important cellular mechanism, conserved from bacteria to humans, responsible for eliminating multiple types of structurally distinct DNA lesions from the genome. The protein XPA appears to play a central role in NER, recognizing and/or verifying damaged DNA and recruiting other proteins, including RPA, ERCC1, and TFIIH, to repair the damage. Sequence analysis and genetic evidence suggest that zinc, which is essential for DNA binding, is associated with a C4-type motif, C-X2-C-X17-C-X2-C. Sequence analysis suggests that a second, H2C2-type zinc-binding motif may be present near the C-terminal. Seventy percent of the amino acid sequence of Xenopus laevis XPA (xXPA) is identical to human XPA and both putative zinc-binding motifs are conserved in all known XPA proteins. Electrospray ionization-mass spectroscopy data show that xXPA contains only one zinc atom per molecule. EXAFS spectra collected on full-length xXPA in frozen (77 K) 15% glycerol aqueous solution unequivocally show that the zinc atom is coordinated to four sulfur atoms with an average Zn--S bond length of 2.33 +/- 0.02 A. Together, the EXAFS and mass spectroscopy data indicate that xXPA contains just one C4-type zinc-binding motif.

Human nucleotide excision repair protein XPA: extended X-ray absorption fine-structure evidence for a metal-binding domain.

The ubiquitous, multi-enzyme, nucleotide excision repair (NER) pathway is responsible for correcting a wide range of chemically and structurally distinct DNA lesions in the eukaryotic genome. Human XPA, a 31 kDa, zinc-associated protein, is thought to play a major NER role in the recognition of damaged DNA and the recruitment of other proteins, including RPA, ERCC1, and TFIIH, to repair the damage. Sequence analyses and genetic evidence suggest that zinc is associated with a C4-type motif, C105-X2-C108-X17-C126-X2-C129, located in the minimal DNA binding region of XPA (M98-F219). The zinc-associated motif is essential for damaged DNA recognition. Extended X-ray absorption fine structure (EXAFS) spectra collected on the zinc associated minimal DNA-binding domain of XPA (ZnXPA-MBD) show directly, for the first time, that the zinc is coordinated to the sulfur atoms of four cysteine residues with an average Zn-S bond length of 2.34+/-0.01 A. XPA-MBD was also expressed in minimal medium supplemented with cobalt nitrate to yield a blue-colored protein that was primarily (>95%) cobalt associated (CoXPA-MBD). EXAFS spectra collected on CoXPA-MBD show that the cobalt is also coordinated to the sulfur atoms of four cysteine residues with an average Co-S bond length of 2.33+/-0.02 A.

Acetylcholine receptors of the Drosophila brain: a 900 bp promoter fragment contains the essential information for specific expression of the ard gene in vivo.

The ard gene encodes a beta-subunit of Drosophila nicotinic acetylcholine receptors specifically expressed in a subset of neurons. To identify the cis-regulatory region responsible for this cell-specific expression, various 5' fragments of the ard gene were fused to a lacZ reporter gene and introduced into the Drosophila genome. A DNA fragment spanning approximately 760 bp upstream and approximately 140 bp downstream of a cluster of putative transcription start sites produced a pattern of beta-galactosidase activity that resembles the distribution of ARD transcripts. Both in embryos and adults the levels of lacZ RNA were similar to those of endogenous ARD transcripts, suggesting that the 900 bp fragment harbors all essential elements for proper expression of the ard gene.

Developmentally regulated alternative splicing of Drosophila myosin heavy chain transcripts: in vivo analysis of an unusual 3' splice site.

The 3' penultimate exon (exon 18) of transcripts from the muscle myosin heavy chain (MHC) gene of Drosophila melanogaster is excluded from mRNAs of embryonic and larval muscle, while it is included in mRNAs of adult thoracic muscles. By transforming organisms with the MHC gene 5' end, linked in-frame to the MHC gene 3' end, we were able to generate correct tissue-specific expression of this minigene and stage-specific splicing of exon 18, indicating that all the cis-acting sequences necessary for alternative splicing are contained within the construct. The 3' splice site that precedes exon 18 is unusually purine-rich, may form a stem-loop structure with the 5' splice site following exon 18, and is conserved relative to the splice site of an alternative exon of the Drosophila alkali myosin light chain gene. We converted the MHC gene 3' splice junction to a consensus splice site and also inserted the branchpoint and 3' splice site of a constitutively spliced intron in its place. These alterations had no effect on the splicing pathway in vivo, ruling out the possibility that the unusual splice junction, or secondary structures that involve this splice junction, directly regulate alternative splicing of exon 18.

Integration into the cellular genome of a Friend murine leukaemia virus containing a selectable marker reveals a clonal origin of erythroleukaemia.

We have constructed a selectable Friend murine leukaemia virus (F-MuLV) with a suppressor tRNA (supF) gene integrated into the proviral long terminal repeat. The viral construct was infectious and pathogenic and retained the marker gene when growing in vitro or in vivo. Only a few integration sites of the provirus were detected by Southern blot analysis of the DNA of erythroleukaemic cells. These results indicate that F-MuLV-induced erythroleukaemia is of clonal origin and suggest that insertional mutagenesis is involved in pathogenesis.

Autocrine stimulation after transfer of the granulocyte/macrophage colony-stimulating factor gene and autonomous growth are distinct but interdependent steps in the oncogenic pathway.

Autocrine stimulation of cells by aberrant synthesis of growth factor may lead to malignant transformation, either as a direct consequence of endogenous factor production or as a first step of a series of successive events. Introduction of the granulocyte/macrophage colony-stimulating factor (GM-CSF) cDNA clone into a vector based on the myeloproliferative sarcoma virus allowed efficient transfer and expression of GM-CSF in factor-dependent myeloid cell lines (FDC-P1 and FDC-P2). Factor-independent growth was acquired when the vector was introduced into the GM-CSF-responsive FDC-P1 cell line but not the multi-CSF-dependent FDC-P2 line. Nonlinear clonability in the absence of exogenous growth factor and growth inhibition by GM-CSF antiserum support a model of autocrine stimulation that requires interaction of factor and receptor at the outer membrane. However, many, but not all, infected FDC-P1 cells acquired subsequently a second mutation that abrogated the requirement of GM-CSF secretion and external interaction. The nature of the second step, which presumably leads to tumorigenicity of these cells, is not well understood, but its frequency could be correlated with the level of GM-CSF released by an individual cell clone.

King Lear and some anxieties of old age.

The experience of ageing and the resulting emotional difficulties of elderly patients have only recently begun to be understood from a psychodynamic perspective. This paper draws on two sources, literary (Shakespeare's King Lear) and clinical (consultations with elderly patients in primary care) to highlight a fundamental anxiety of old age: the dread of being abandoned to a state of utter helplessness. This state is felt to be caused by the catastrophes of old age, such as stroke and dementia. The defence against this anxiety is often a narcissistic tyranny.