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At least 109 records · Page 6Linked to original sources
PubMed2003

Natural killer cell receptor signaling.

Natural killer (NK) cell immune responses are regulated by a balance of activating and inhibitory signals transmitted by cell surface receptors. Immunoreceptor tyrosine-based inhibition motifs in the cytoplasmic domains of inhibitory NK receptors recruit tyrosine or lipid phosphatases, which modulate the activation signals transmitted by receptors linked to the Syk and ZAP70 tyrosine kinases and phosphatidylinositol-3 kinases. In addition, recent studies of gene-deficient animals, in particular Syk and ZAP70 double-deficient mice, suggest that NK cells possess a robust and potentially redundant receptor system to ensure their development and function.

Animals↗
PubMed2002

Establishing through-bond connectivity in solids with NMR: structure and dynamics in HC(60)(+).

We present a novel nuclear magnetic resonance experiment for establishing through-bond connectivity in solids using scalar coupling-driven correlation. This method, a variant of the popular double-quantum-filtered correlation spectroscopy experiment in liquids, is robust under fast magic-angle-spinning conditions and in the presence of dynamics. In HC(60)(+), where anisotropic molecular motion renders through-space dipolar-driven correlation ineffective, this through-bond correlation method answers a significant structural question by accurately identifying the direct bond between the protonated sp(3) hybridized carbon site and the sp(2) hybridized cationic site.

Magnetic Resonance Spectroscopy↗
PubMed2003

Through-bond 13C-13C correlation at the natural abundance level: refining dynamic regions in the crystal structure of vitamin-D3 with solid-state NMR.

Recently, we presented a novel nuclear magnetic resonance experiment for establishing through-bond connectivity in disordered solids using scalar coupling-driven correlation. This method, a variant of the popular double-quantum-filtered correlation spectroscopy experiment in liquids, is robust under fast magic-angle-spinning conditions and in the presence of dynamics. Here, we show that this new experiment, the UC2QF COSY, can be extended to 13C natural abundance correlation in moderately sized molecules, allowing the assignment of the 54 peaks of the solid-state NMR spectrum of microcrystalline vitamin-D3. In this case, comparison between the assigned peaks and ab initio calculations of the chemical shifts based on the crystal coordinates permits a refinement of the average structure in dynamic regions reported as disordered in the crystal structure.

Carbon Isotopes↗
PubMed1999

Dimerization of the calcium-sensing receptor occurs within the extracellular domain and is eliminated by Cys --> Ser mutations at Cys101 and Cys236.

Calcium-sensing receptors are present in membranes as dimers that can be reduced to monomers with sufhydryl reagents. All studies were carried out on the human calcium-sensing receptor tagged at the carboxyl terminus with green fluorescent protein (hCaR-GFP) to permit identification and localization of expressed proteins. Truncations containing either the extracellular agonist binding domain plus transmembrane helix 1 (ECD/TMH1-GFP) or the transmembrane domain plus the intracellular carboxyl terminus (TMD/carboxyl terminus-GFP) were used to identify the dimerization domain. ECD/TMH1-GFP was a dimer in the absence of reducing reagents, whereas TMD/carboxyl-terminal GFP was a monomer in the absence or presence of reducing agents, suggesting that dimerization occurs via the ECD. To identify the residue(s) involved in dimerization within the ECD, cysteine --> serine point mutations were made in residues that are conserved between hCaR and metabotropic glutamate receptors. Mutations at positions 60 and 131 were expressed at levels comparable to wild type in HEK 293 cells, had minimal effects on hCaR function, and did not eliminate dimerization, whereas mutations at positions 101 and 236 greatly decreased receptor expression and resulted in significant amounts of monomer in the absence of reducing agents. The double point mutant hCaR(C101S/C236S)-GFP was expressed more robustly than either C101S or C236S and covalent dimerization was eliminated. hCaR(C101S/C236S)-GFP had a decreased affinity for extracellular Ca2+ and slower response kinetics upon increases or decreases in agonist concentration. These results suggest that covalent, disulfide bond-mediated dimerization of the calcium-sensing receptor contributes to stabilization of the ECD and to acceleration of the transitions between inactive and active receptor conformations.

Base Sequence↗
PubMed2003

Fission yeast Tup1-like repressors repress chromatin remodeling at the fbp1+ promoter and the ade6-M26 recombination hotspot.

Chromatin remodeling plays crucial roles in the regulation of gene expression and recombination. Transcription of the fission yeast fbp1(+) gene and recombination at the meiotic recombination hotspot ade6-M26 (M26) are both regulated by cAMP responsive element (CRE)-like sequences and the CREB/ATF-type transcription factor Atf1*Pcr1. The Tup11 and Tup12 proteins, the fission yeast counterparts of the Saccharomyces cerevisiae Tup1 corepressor, are involved in glucose repression of the fbp1(+) transcription. We have analyzed roles of the Tup1-like corepressors in chromatin regulation around the fbp1(+) promoter and the M26 hotspot. We found that the chromatin structure around two regulatory elements for fbp1(+) was remodeled under derepressed conditions in concert with the robust activation of fbp1(+) transcription. Strains with tup11delta tup12delta double deletions grown in repressed conditions exhibited the chromatin state associated with wild-type cells grown in derepressed conditions. Interestingly, deletion of rst2(+), encoding a transcription factor controlled by the cAMP-dependent kinase, alleviated the tup11delta tup12delta defects in chromatin regulation but not in transcription repression. The chromatin at the M26 site in mitotic cultures of a tup11delta tup12delta mutant resembled that of wild-type meiotic cells. These observations suggest that these fission yeast Tup1-like corepressors repress chromatin remodeling at CRE-related sequences and that Rst2 antagonizes this function.

Chromatin↗
PubMed2000

Pseudorabies virus membrane proteins gI and gE facilitate anterograde spread of infection in projection-specific neurons in the rat.

The membrane proteins gI and gE of Pseudorabies virus (PRV) are required for viral invasion and spread through some neural pathways of the rodent central nervous system. Following infection of the rat retina with wild-type PRV, virus replicates in retinal ganglion neurons and anterogradely spreads to infect all visual centers in the brain. By contrast, gI and gE null mutants do not infect a specific subset of the visual centers, e.g., the superior colliculus and the dorsal lateral geniculate nucleus. In previous experiments, we suggested that the defect was not due to inability to infect projection-specific retinal ganglion cells, because mixed infection of a gE deletion mutant and a gI deletion mutant restored the wild-type phenotype (i.e., genetic complementation occurred). In the present study, we provide direct evidence that gE and gI function to promote the spread of infection after entry into primary neurons. We used stereotaxic central nervous system injection of a fluorescent retrograde tracer into the superior colliculus and subsequent inoculation of a PRV gI-gE double null mutant into the eye of the same animal to demonstrate that viral antigen and fluorescent tracer colocalize in retinal ganglion cells. Furthermore, we demonstrate that direct injection of a PRV gI-gE double null mutant into the superior colliculus resulted in robust infection followed by retrograde transport to the eye and replication in retinal ganglion neuron cell bodies. These experiments provide additional proof that the retinal ganglion cells projecting to the superior colliculus are susceptible and permissive to gE and gI mutant viruses. Our studies confirm that gI and gE specifically facilitate anterograde spread of infection by affecting intracellular processes in the primary infected neuron such as anterograde transport in axons or egress from axon terminals.

Animals↗
PubMed1999

Benzo-pyrones in the treatment of lymphoedema.

Fifty clinical trials of 4 benzo-pyrones in the treatment of lymphoedema, by over 37 authors in 8 countries, are reviewed: 38 oral and 12 topical (11 and 6 of these, added to other therapies). Oral benzo-pyrones reduced oedema, symptoms (in almost all) and inflammation (SAI). These were significant and clinically important. There were no significant differences between arms and Grades 1 and 2 legs. Combining these 20 trials gave mean annual reductions of 55% of oedema (SE: 7.8%; 95% Confidence Interval: 40% to 71%) (p<0.001). Four trials of elephantitic legs gave 17% (4.8%; 7.6% to 27%), significantly less (p<0.01). Meta-analyses, tested by omitting non-double-blind or non-peer-reviewed trials, were robust. The greater the oedema, the greater the rate of reduction-lessening as time passed and the oedema reduced: annual reduction=37%x(79%) Period (p=0.01). Reductions varied with the molar dose (p=10(-8)): =0.10% (SE 0.013%) Dose (mg of coumarin or molar equivalent of other drugs). Topical coumarin also reduced oedema and symptoms. The results of some other therapies were improved by oral or topical benzo-pyrones 15% to 22% over a month and 0% to 78% over a year. These drugs are slow, but effective, cheap and convenient. Because of their slowness, compression garments are unnecessary. They were seldom used in trials. Side-effects are minimal. Only oral coumarin may cause idiosyncratic hepatitis (3 per 1,000). Topical coumarin does not, nor other benzo-pyrones.

Administration, Oral↗
PubMed2004

Meiosis-specific yeast Hop1 protein promotes synapsis of double-stranded DNA helices via the formation of guanine quartets.

In most eukaryotes, genetic exchange between paired homologs occurs in the context of a tripartite proteinaceous structure called the synaptonemal complex (SC). Genetic analyses have revealed that the genes encoding SC proteins are vital for meiotic chromosome pairing and recombination. However, the number, nature and/or the mechanism used by SC proteins to align chromosomes are yet to be clearly defined. Here, we show that Saccharomyces cerevisiae Hop1, a component of SC, was able to promote pairing of double-stranded DNA helices containing arrays of mismatched G/G sequences. Significantly, pairing was rapid and robust, independent of homology in the arms flanking the central G/G region, and required four contiguous guanine residues. Furthermore, data from truncated DNA double helices showed that 20 bp on either side of the 8 bp mismatched G/G region was essential for efficient synapsis. Methylation interference indicated that pairing between the two DNA double helices involves G quartets. These results suggest that Hop1 is likely to play a direct role in meiotic chromosome pairing and recombination by its ability to promote synapsis between double-stranded DNA helices containing arrays of G residues. To our knowledge, Hop1 is the first protein shown to promote synapsis of DNA double helices from yeast or any other organism.

Base Pair Mismatch↗
PubMed2002

Pathogenic effects of cerebral amyloid angiopathy mutations in the amyloid beta-protein precursor.

Cerebral amyloid beta-protein angiopathy (CAA) is a key pathological feature of patients with Alzheimer's disease and certain related disorders. Several mutations have been identified within the Abeta region of the Abeta protein precursor (AbetaPP) gene that appear to enhance the severity of CAA. A new mutation has been identified within the Abeta region (D23N) of AbetaPP that is associated with severe CAA in an Iowa kindred. Recently, we showed that E22Q Dutch, D23N Iowa, and E22Q/D23N Dutch/Iowa double-mutant Abeta40 peptides rapidly assemble in solution to form fibrils compared to wild-type Abeta40. Similarly, the E22Q Dutch and D23N Iowa Abeta40 peptides were found to induce robust pathologic responses in cultured human cerebrovascular smooth muscle (HCSM) cells, including elevated levels of cell-associated AbetaPP, proteolytic breakdown of actin, and cell death. Double-mutant E22Q/D23N Dutch/Iowa Abeta40 was more potent than either single-mutant form of Abeta in causing pathologic responses in HCSM cells. These in vitro data suggested that the E22Q Dutch and D23N Iowa substitutions promote fibrillogenesis and the pathogenicity of Abeta towards HCSM cells. Moreover, the presence of both CAA substitutions in the same Abeta peptide further enhances the fibrillogenic and pathogenic properties of Abeta. We also have generated transgenic mouse models to examine the effects of single and double CAA mutations in AbetaPP in vivo. Preliminary analysis of transgenic mouse brains indicates that expression of double-mutant E22Q/D23N Dutch/Iowa AbetaPP leads to robust deposition of Abeta in a vascular-weighted manner.

Amino Acid Sequence↗
PubMed2004

Simulation based analysis of automated, classification of medical images.

OBJECTIVES: The ability of various classifiers to discriminate between normal and glaucomatous eyes based on features derived from automated analysis of laser scanning images of the eye background is investigated. METHODS: To compare the classifiers without over-optimization for a given dataset, we use a simulation model to create topography images. We designed three different simulation setups as model of extreme situations and medical subgroups. RESULTS: Neither linear nor tree-based classifiers are ideal for all setups. The most robust performance is obtained by a combination of both, so-called Double-Bagging. Classification of real data from a case-control study shows best results with Double-Bagging. All results obtained with the analysis method extracting features automatically are worse than those obtained by the same classifiers but with features derived from an analysis method that requires intervention of a physician. CONCLUSIONS: Robust classification results for classification of laser scanning images obtained with the Heidelberg Retina Tomograph are achieved by combined classifiers. The examined automated procedure causes an increased misclassification error compared to the established clinical routine requiring an expert physician's intervention.

Automation↗
PubMed2006

Attention-deficit/hyperactivity disorder: use of cognitive evoked potential (P300) to predict treatment response.

OBJECTIVE: To evaluate the use of P300 in predicting treatment response to medicines in patients with Attention-Deficit/Hyperactivity Disorder (ADHD), and to confirm previous reports that 31-electrode mean auditory P300 amplitude (AA) predicts response to atomoxetine; and right fronto-central to parietal AA ratio predicts response to methylphenidate. METHODS: Efficacy and P300 data from 58 children with ADHD enrolled in a double-blind crossover study using atomoxetine and methylphenidate were analyzed. Robust response was defined as 60% decrease from baseline in the ADHD rating scale. Response was alternately defined as greater than 50% decrease. RESULTS: Pre-treatment mean 31-electrode AA>6.8 microV predicted response to atomoxetine using both definitions of response. Right fronto-central to parietal AA ratio did not predict response to methylphenidate. A previous report that methylphenidate responders differed from non-responders in pre-treatment AA at T8 was confirmed, and AA at T8>7.65 microV predicted response to methylphenidate. 31-electrode mean P300 visual latency (VL) also predicted response to atomoxetine, as previously reported with imipramine. CONCLUSIONS: Mean AA predicts response to atomoxetine in ADHD patients. AA at T8 predicts response to methylphenidate. Such predictive tools may allow individually tailored choice of medicine in treatment of ADHD. SIGNIFICANCE: This allows a more informed decision of which medicine to use for a given patient.

Acoustic Stimulation↗
PubMed1998

Easy-to-implement method to target nonlinear systems.

In this work we present a method to rapidly direct a chaotic system, to an aimed state or target, through a sequence of control perturbations, with few different amplitudes chosen according to the allowed control-parameter changes. We applied this procedure to the one-dimensional Logistic map, to the two-dimensional Henon map, and to the Double Scroll circuit described by a three-dimensional system of differential equations. Furthermore, for the Logistic map, we show numerically that the resulting trajectory (from the starting point to the target) goes along a stable manifold of the target. Moreover, using the Henon map, we create and stabilize unstable periodic orbits, and also verify the procedure robustness in the presence of noise. We apply our method to the Double Scroll circuit, without using any low-dimensional mapping to represent its dynamics, an improvement with respect to previous targeting methods only applied for experimental systems that are mapping-modeled. (c) 1998 American Institute of Physics.

Journal Article↗
PubMed1998

[Principles of energy sources of totally implantable hearing aids for inner ear hearing loss].

A fully implantable hearing aid consists of a sound receptor (microphone), an electronic amplifier including active audio-signal processing, an electromechanical transducer (actuator) for stimulating the ear by vibration, and an energy source. The energy source may be either a primary cell or a rechargeable (secondary) cell. As the energy requirements of an implantable hearing aid are dependent on the operating principle of the actuator, the operating principles of electromagnetic and piezoelectric transducers were examined with respect to their relative power consumption. The analysis showed that the energy requirements of an implantable hearing aid are significantly increased when an electromagnetic transducer is used. The power consumption of a piezoelectric transducer was found to be less than that of the electronic components alone. The energy needed to run a fully implantable hearing aid under these conditions would be 38 mWH per day. Primary cells cannot provide the energy needed for a minimum operation time of 5 years (70 WH), and therefore rechargeable cells must be used. A theoretical appraisal was carried out on nickel-cadmium, nickel-metal hydride, and lithium-ion cells to determine their suitability as well as to assess the risks associated with their use in an implant. Safety measures were drawn up from the results. Ni-MH cells were found to be the most suitable for use as an energy source for implantable hearing-aids because they are more robust than Li ion cells and their storage capacity is double that of Ni-Cd cells of similar size.

Cochlear Implants↗
PubMed2004

Prediction of the reproductive status of cattle on the basis of milk progesterone measures: model description.

Reproductive management, in particular timely oestrus detection, is important for profitable dairy production. The aim of this study was to develop a biological model to predict reproductive state on the basis of milk progesterone measures. A number of additional inputs were incorporated to make use of other known effectors of reproductive performance that are not reflected in progesterone levels. These are: days from calving, breed, parity, signs of behavioural oestrus, insemination dates, pregnancy determinations, energy status, body fat status, milk urea content and reproductive disorders associated with calving. A dynamic, deterministic model was developed. It is designed to run each time a new trigger input (progesterone, behavioural oestrus, inseminations, pregnancy determinations) occurs using the current and previous values and can run in the absence of the additional inputs. The milk progesterone values are smoothed using an extended Kalman filter before being processed in the biological component of the model. The model predicts the reproductive status of the cow, which can be one of three mutually exclusive states: postpartum anoestrus, oestrus cycling, and potentially pregnant. The other model outputs are all reproductive status specific with the exception of days to next sample (DNS), which is calculated in each model run regardless of reproductive status. DNS is designed to feedback to the sampling system so that the frequency of milk sampling (i.e. progesterone measurement) can be varied according to the predicted likelihood of a future reproductive event, such as onset of oestrus cycling. The other model outputs are: risk of prolonged postpartum anoestrus, risk and type of ovarian cyst, onset of oestrus, likelihood of a potential insemination succeeding, and likelihood of being pregnant (following oestrus). The model was evaluated using three simulated datasets consisting of a timeseries of progesterone values centred on each of the three reproductive statuses and including relevant additional information. Test runs were carried out on the full datasets and then on reduced data. The data reductions were made by using only those values that would have been available if the model days to next sample function was used to control sampling frequency. The sensitivity of the model to noise in the raw progesterone data was examined by adding 1, 2, or 3 residual standard deviations (1.85 ng/ml) random variation to the original data and evaluating model performance. The model was found to be able to readily identify and distinguish reproductive states. A reduction in sampling frequency to 36% of original sample resulted in an average increase in days to detection of oestrus of 0.36. The addition of 1 S.D. noise did not cause additional oestruses to be detected and all oestruses were correctly identified. However, when 2 or 3 S.D. noise were added, the model found on average 1.4 and 3 extra oestruses. It was concluded that reproductive status can be predicted from milk progesterone values using a biological model and that such a model is robust to reductions in sampling frequency number and to a doubling in the random variation in the raw progesterone values. It therefore has the potential to provide the basis for a useful reproductive management tool.

Anestrus↗
PubMed2005

Improving outcomes in heart transplantation: the potential of proliferation signal inhibitors.

Graft failure and mortality among heart transplant recipients remains higher than in populations receiving renal transplants. A major cause of graft loss is cardiac allograft vasculopathy (CAV), a condition characterized by diffuse thickening of coronary blood vessels. CAV often progresses silently, with major cardiac events (eg, ventricular arrhythmia) being the first presentation. Better diagnosis and monitoring of CAV is now possible with intravascular ultrasonography, a sensitive technique for measuring intimal thickness. To date, immunosuppressants have shown little efficacy for preventing CAV. However, a new class of agents, proliferation signal inhibitors (sirolimus and everolimus), have shown considerable efficacy in this regard and for preventing rejection. In an open-label trial, sirolimus therapy was associated with less intimal and medial proliferation than azathioprine. More robust evidence is available from a larger-scale, double-blind trial involving everolimus. At 12-month follow-up the incidence of CAV was significantly lower in patients receiving everolimus (35.7% and 30.4% for everolimus 1.5 and 3.0 mg/d vs 52.8% for azathioprine; P < .05). Sirolimus and everolimus were also associated with a lower rate of cytomegalovirus infection. As with other immunosuppressants, these agents are associated with adverse events (eg, hyperlipidemia), but they can be managed. Coadministration with calcineurin inhibitors (CNIs) can exacerbate CNI-related nephrotoxicity, but evidence suggests that everolimus administered with reduced-exposure cyclosporine in the maintenance phase preserves renal function without loss of immunosuppressive efficacy. Reduced CNI dosing in de novo patients is also a potential future benefit. Proliferation signal inhibitors have considerable potential for improving outcomes in heart transplantation.

Cell Division↗
PubMed2004

The 40-mg dose of eletriptan: comparative efficacy and tolerability versus sumatriptan 100 mg.

Meta-analysis provides valuable information regarding relative efficacies of triptans, but head-to-head comparator studies remain the gold standard. Three similar head-to-head trials comparing eletriptan 40 mg (E40) with sumatriptan 100 mg (S100) provide a rare opportunity and sufficient power, for robust comparisons of efficacy. Data were combined from three double-blind, placebo-controlled, first-dose, first-attack acute migraine treatment studies comparing E40 (n=1132), S100 (n=1129), and placebo (n=645). The primary outcome was headache response at 2 h. Secondary outcomes included headache response at 1 h, pain-free and functional responses, and sustained headache and pain-free responses. Odds ratios were calculated for summary estimates of probability of response. There were higher headache response rates with eletriptan versus sumatriptan at 2 h (67% vs. 57%; P<0.0001) and 1 h (34% vs. 26%; P<0.0001). Eletriptan also had higher 2 h pain-free (35% vs. 25%; P<0.0001) and functional responses (67% vs. 58%; P<0.0001). Sustained headache (42%) and pain-free (22%) response rates were higher for eletriptan versus sumatriptan (34%, P<0.0001; 15%, P<0.0001). The probability of response for eletriptan versus sumatriptan ranged from 36% higher (relief of nausea) to 64% higher (sustained pain-free rate). Combined analysis demonstrates that E40 has superior efficacy versus S100 across all clinically relevant outcomes.

Adolescent↗
PubMed1999

The RAD2 domain of human exonuclease 1 exhibits 5' to 3' exonuclease and flap structure-specific endonuclease activities.

The RAD2 family of nucleases includes human XPG (Class I), FEN1 (Class II), and HEX1/hEXO1 (Class III) products gene. These proteins exhibit a blend of substrate specific exo- and endonuclease activities and contribute to repair, recombination, and/or replication. To date, the substrate preferences of the EXO1-like Class III proteins have not been thoroughly defined. We report here that the RAD2 domain of human exonuclease 1 (HEX1-N2) exhibits both a robust 5' to 3' exonuclease activity on single- and double-stranded DNA substrates as well as a flap structure-specific endonuclease activity but does not show specific endonuclease activity at 10-base pair bubble-like structures, G:T mismatches, or uracil residues. Both the 5' to 3' exonuclease and flap endonuclease activities require a divalent metal cofactor, with Mg(2+) being the preferred metal ion. HEX1-N2 is approximately 3-fold less active in Mn(2+)-containing buffers and exhibits <5% activity in the presence of Co(2+), Zn(2+), or Ca(2+). The optimal pH range for the nuclease activities of HEX1-N2 is 7.2-8.2. The specific activity of its 5' to 3' exonuclease function is 2.5-7-fold higher on blunt end and 5'-recessed double-stranded DNA substrates compared with duplex 5'-overhang or single-stranded DNAs. The flap endonuclease activity of HEX1-N2 is similar to that of human flap endonuclease-1, both in terms of turnover efficiency (k(cat)) and site of incision, and is as efficient (k(cat)/K(m)) as its exonuclease function. The nuclease activities of HEX1-N2 described here indicate functions for the EXO1-like proteins in replication, repair, and/or recombination that may overlap with human flap endonuclease-1.

Amino Acid Sequence↗
PubMed2006

The 3' untranslated regions of Kamiti River virus and Cell fusing agent virus originated by self-duplication.

Previously, it was shown that the 3' untranslated region (3'UTR) of Kamiti River virus (KRV) is nearly twice as long as the 3'UTR of other flaviviruses (1208 nucleotides compared with 730 nucleotides for the longest 3'UTR of any virus in the Tick-borne encephalitis virus species). Additionally, KRV and the closely related Cell fusing agent virus (CFAV) were shown to contain two short, almost perfect repeat sequences of 67 nucleotides. However, the construction of a robust comparative nucleotide alignment has now revealed that the double-length 3'UTR and the direct repeats resulted from the virtually complete duplication of a primordial KRV 3'UTR. We also propose that the CFAV 3'UTR was derived from a KRV-like precursor sequence with a large deletion that nevertheless preserved the two direct repeat sequences. These data provide new insights into the evolution of the flavivirus 3'UTR.

3' Untranslated Regions↗