Arrhythmia discrimination using hemoglobin spectroscopy in humans.

BACKGROUND: Inappropriate therapies are frequently delivered by implantable cardioverter-defibrillators (ICDs). We have investigated muscle perfusion as a means of augmenting arrhythmia discrimination by using implanted near-infrared spectroscopy. OBJECTIVE: To evaluate hemodynamic stability by monitoring muscle perfusion from within the ICD pocket, in fresh tissue and inside the scar capsule on preexisting ICD generators, during induced cardiac arrhythmias, in humans. METHODS: The sensor was implanted on or under the pectoral muscle, during ICD defibrillation threshold testing. A microvascular oxygenation trend indicator (O2 Index) was computed during 74 induced ventricular fibrillation and 34 normal sinus rhythm episodes in 34 patients and also during 28 atrial and 90 ventricular overdrive pacing episodes as simulations of supraventricular and ventricular tachycardias, respectively. RESULTS: On average, the change in oxygenation, based on the O2 Index, was statistically significant (P <.003) from baseline within 3 seconds following cardiac arrest. An optimized O2 Index, used for detecting the hemodynamic trend, exhibited a decreasing trend during ventricular fibrillation (P <.0001) and was different from that during normal sinus rhythm (P <.0001). The sensitivity for the detection of ventricular fibrillation was 100%, and the specificity for the rejection of normal sinus rhythm was 82% in the presence of scar tissue on the optical sensor. For a 35-mm Hg drop in the mean arterial pressure as the threshold for hemodynamic instability, the specificity for the rejection of hemodynamically stable atrial and ventricular pacing episodes was 93% and 71%, respectively. CONCLUSION: An implantable near-infrared spectroscopic sensor may be useful for hemodynamic monitoring during cardiac arrhythmias to prevent inappropriate therapy.

Comparison of Muscodor albus Volatiles with a Biorational Mixture for Control of Seedling Diseases of Sugar Beet and Root-Knot Nematode on Tomato.

A biorational synthetic mixture of organic components mimicking key antimicrobial gases produced by Muscodor albus was equivalent to the use of live M. albus for control of seedling diseases of sugar beet (Beta vulgaris) caused by Pythium ultimum, Rhizoctonia solani AG 2-2, and Aphanomyces cochlioides. The biorational mixture provided better control than the live M. albus formulation for control of root-knot nematode, Meloidogyne incognita, on tomato (Lycopersicon esculentum). The biorational mixture provided control of damping-off equal to a starch-based formulation of the live fungus for all three sugar beet pathogens, and significantly reduced the number of root-knot galls on tomato roots compared with a barley-based formulation. Rate studies with the biorational mixture showed that 2 and 0.75 µl/cm3 of soil were required to provide optimal control of Rhizoctonia and Pythium damping-off of sugar beet, respectively. Five microliters of biorational mixture per milliliter of water was required for 100% mortality in 24 h for Meloidogyne incognita in in vitro studies. In in vivo studies, 1.67 µl of the biorational mixture/cm3 of sand resulted in fewer root-knot galls than a Muscodor albus infested ground barley formulation applied at 5 g/liter of sand.

Mycofumigation with Muscodor albus and Muscodor roseus for Control of Seedling Diseases of Sugar Beet and Verticillium Wilt of Eggplant.

Mycofumigation is the use of antimicrobial volatiles produced by fungi such as Muscodor albusitalic and M. roseus for the control of other organisms. Sugar beet (Beta vulgaris L.) stand establishment was increased and disease severity decreased by mycofumigation with M. roseus and M. albus in autoclaved soil infested with Rhizoctonia solani, Pythium ultimum, or Aphanomyces cochlioides. Eggplant seedlings (Solanum melongena L.) transplanted into autoclaved soil infested with Verticillium dahliae and mycofumigated with M. albus and M. roseus had significantly less disease (P < 0.05) after 4 and 5 weeks compared with nonmycofumigated Verticillium-infested soil. The effect of formulation on efficacy of mycofumigation with M. roseus was tested using potato dextrose agar strips, alginate capsules, ground barley, pesta granules, and stabileze granules. The stabileze and ground barley formulations of M. roseus resulted in the best control of P. ultimum damping-off. The best control of A. cochlioides damping-off was with the stabileze formulation, and the stabileze, ground barley, and agar strip formulations provided similar control of R. solani damping-off. In soil infested with P. ultimum, mycofumigation with M. albus stabileze formulation resulted in stand establishment similar to that in the autoclaved soil. Mycofumigation was ineffective in controlling Fusarium wilt of sugar beet. Neither M. albus nor M. roseus affected sugar beet or eggplant growth or appearance except in the stabileze formulation, where stunting was noticed. Mycofumigation with M. albus and M. roseus shows promise for control of soilborne diseases caused by P. ultimum, A. cochlioides, R. solani, and V. dahliae.

Two parameters improve efficiency of mitochondrial uptake of adenylate kinase: decreased folding velocity and increased propensity of N-terminal alpha-helix formation.

The long isoform of eukaryotic adenylate kinase has a dual subcellular location in the cytoplasm and in the mitochondrial intermembrane space. Protein sequences and modifications are identical in both locations. In yeast, the bulk of the major form of adenylate kinase (Aky2p) is in the cytoplasm and, in the steady state, only 5-8% is sorted to the mitochondrial intermembrane space. Since the reasons for exclusion from mitochondrial import are unclear, we have constructed aky2 mutants with elevated mitochondrial uptake efficiency of Aky2p in vivo and in vitro. We have analyzed the effect of the mutations on secondary structure prediction in silico and have tested folding velocity and folding stability. One type of mutants displayed decreased proteolytic stability and retarded renaturation kinetics after chaotropic denaturation implying that deterioration of folding leads to prolonged presentation of target information to mitochondrial import receptors, thereby effecting improved uptake. In a second type of mutants, increased import efficiency was correlated with an increased probability of formation of an alpha-helix with increased amphipathic moment at the N-terminus suggesting that targeting interactions with mitochondrial import receptors had been improved at the level of binding affinity.

Volatile antimicrobials from Muscodor albus, a novel endophytic fungus.

Muscodor albus is a recently described endophytic fungus obtained from small limbs of Cinnamomum zeylanicum (cinnamon tree). This xylariaceaous fungus effectively inhibits and kills certain other fungi, and bacteria, by virtue of a mixture of volatile compounds that it produces. The majority of these compounds were identified by gas chromatography/mass spectrometry and then made into an artificial mixture that mimicked the antibiotic effects of the mixture of volatile compounds given off by the fungus. Each of the five classes of volatile compounds produced by the fungus (alcohols, esters, ketones, acids and lipids) had some inhibitory effect against the test fungi and bacteria, but none was lethal. However, collectively they acted synergistically to kill a broad range of plant- and human-pathogenic fungi and bacteria. The most effective class of inhibitory compounds was the esters, of which 1-butanol, 3-methyl-, acetate was the most active biologically. This report describes the ecological implications and potential practical benefits of the 'mycofumigation' effects of M. albus.

Characterization of stereochemistry and molecular conformation using solid-state NMR tensors.

A solid-state NMR technique is described for establishing stereochemistry using the natural product terrein as a model compound. This method involves comparison of experimental (13)C tensor principal values with ab initio computed values for all possible computer-generated stereoisomers. In terrein the relative stereochemistry is confirmed by NMR to be 2R*,3S with high statistical probability (>99.5%). The proposed approach also simultaneously verifies the molecular conformation of the two hydroxy groups in terrein established by X-ray data. It is sufficient to use only shift tensor values at carbons 2 and 3, the stereocenters, to characterize both the stereochemistry and molecular conformations. The solid-state NMR method appears to be especially useful for determining relative stereochemistry of compounds or their derivatives that are difficult to crystallize.

Two telomerase reverse transcriptases (TERTs) expressed in Candida albicans.

The human pathogenic yeast Candida albicans contains two telomerase reverse transcriptase (TERT) genes. CaTERT1 and CaTERT2 appear either to be two alleles of the same gene or two entirely different genes that encode 867-residue proteins that differ by five amino acids. Both TERTs have a calculated pI of 9.5 and a M(r) of 100.9 kDa and are the smallest TERTs identified to date. Both genes appear to be expressed. Based on sequence similarity between CaTERT1 and the Saccharomyces cerevisiae orthologue Est2p, we suggest a revised alignment for motif E of Est2p. The identification of these TERT genes provides the first opportunity to study telomerase in an important human pathogen.

Successful pediatric stenting of a nonthrombotic coronary occlusion as a complication of radiofrequency catheter ablation.

This is a case of a right coronary artery occlusion complicating a RF catheter ablation of a posteroseptal accessory connection in an 8-year-old boy. After multiple balloon angioplasty attempts in the occluded vessel, only transient patency was achieved. The occlusion was successfully treated with placement of an intracoronary stent.

Evidence for paclitaxel from three new endophytic fungi of Himalayan yew of Nepal.

Taxus wallichiana, collected from the Himalayan region of Nepal, yielded, among others, three different endophytic fungi including Sporormia minima, Trichothecium sp., and an unidentified dimorphic fungus. Each was shown to produce paclitaxel in a culture medium. Each of these fungi represents a new report as a paclitaxel producer. The production of paclitaxel was confirmed by an immunoassay that utilized monoclonal antibodies, TLC chromatography as well as high performance liquid chromatography combined with on-line mass spectrometry.

Exploring chemical diversity of epoxyquinoid natural products: synthesis and biological activity of (-)-jesterone and related molecules.

Enantioselective syntheses of the potent antifungal agent (-)-jesterone, its hydroxy epimer, and a dimeric quinone epoxide derivative are reported. The synthesis involves diastereoselective epoxidation of a chiral quinone monoketal derivative and regio- and stereoselective reduction of a quinone epoxide intermediate.

Carnitine/acylcarnitine translocase deficiency (neonatal phenotype): successful prenatal and postmortem diagnosis associated with a novel mutation in a single family.

The neonatal phenotype of carnitine-acylcarnitine translocase (CACT) deficiency is one of the most severe and usually lethal mitochondrial fat oxidation disorders characterized by hypoketotic hypoglycemia, hyperammonemia, cardiac abnormalities, and early death. In this study, the proband was the daughter of consanguineous Hispanic parents. At 36 h of life, she had bradycardia and died at 4 days of age without a specific diagnosis. In a subsequent pregnancy, prenatal counseling and amniocentesis were provided. Incubation of the amniocytes from this pregnancy and fibroblasts (from the dead proband) with [16-(2)H(3)]palmitic acid and analysis by tandem mass spectrometry revealed an increasedconcentration of [16-(2)H(3)]palmitoylcarnitine, suggesting the diagnoses of either CACT or carnitine palmitoyltransferase II (CPT-II) deficiency. CACT enzyme activity was absent in both cell lines. Molecular investigation of cDNA from the dead proband and her affected sibling revealed aberrant CACT cDNA species, including exon 3 skipping, both exon 3 and 4 skipping, and a 13-bp insertion at cDNA position 388. Investigation of these cell lines for mutations affecting CACT RNA processing by analysis of CACT gene sequences, including intron and exon boundaries, revealed a single nucleotide G deletion at the donor site in intron 3 which resulted in exon skipping and a 13-bp insertion. The proband and her affected sibling were homozygous for this deletion.

Jesterone and hydroxy-jesterone antioomycete cyclohexenone epoxides from the endophytic fungus Pestalotiopsis jesteri.

Jesterone and hydroxy-jesterone are novel highly functionalized cyclohexenone epoxides isolated from a newly described endophytic fungal species--Pestalotiopsis jesteri. They were purified from cultures of the fungus by bioassay guided fractionation using Pythium ultimum as the indicator organism. Jesterone, in particular, displays selective antimycotic activity against the oomycetous fungi which are some of the most plant pathogenic of all disease causing fungi. The possible importance of these cyclohexenones to the biology of the endophytic fungus-host plant relationship is also discussed.

Ambuic acid, a highly functionalized cyclohexenone with antifungal activity from Pestalotiopsis spp. and Monochaetia sp.

Ambuic acid, a highly functionalized cyclohexenone, was isolated and characterized from Pestalotiopsis spp. and Monochaetia sp. these being biologically related endophytic fungi associated with many tropical plant species. This compound was found in representative isolates of these fungal species obtained from rainforest plants located on several continents. The relevance of ambuic acid to the biology of the association of these fungi to their host plants is also discussed.

An antiproliferative norditerpene dilactone, Nagilactone C, from Podocarpus neriifolius.

An ethanolic extract of Podocarpus neriifolius D. Don (Podocarpaceae) showed antiproliferative activity against two major tumor cell lines, viz. human HT-1080 fibrosarcoma and murine color 26-L5 carcinoma. Bioassay guided fractionation showed the highest antiproliferative activity in chloroform-soluble fraction. Nagilactone C, the major constituent of this fraction was isolated and characterized by using NMR, IR and FAB-MS spectroscopic methods. Nagilactone C possessed potent antiproliferative activity against human fibrosarcoma and murine colon carcinoma tumor cell lines exhibiting ED50 values of 2.3 and 1.2 microg/ml, respectively. Hence, nagilactone C could be the active constituent present in this plant.

Stable plasma membrane expression of the soluble domain of the human insulin receptor in yeast.

The soluble cytoplasmic kinase domain of the human insulin receptor was N-terminally equipped with either an N-acetylation or a dual-acylation motif (MGC box, to allow myristoylation/palmitoylation) and expressed in yeast cells under the control of the inducible CUP1 promoter. Although the cellular concentration was about the same in both instances (reflecting similar stability against proteolysis), only the myristoylated protein was capable of autophosphorylation to a significant extent and was active to phosphorylate endogenous yeast proteins at tyrosine residues in vivo. Cellular subfractionation showed that the insulin receptor was associated with plasma membranes, from where it was not extractable with high salt or alkali, but a significant fraction was also localized in the nuclear fraction. The myristoylated protein is absent from the cytoplasm. No effect of expression of either the acetylated or the myristoylated version on growth and respiration on various carbon sources was detected, suggesting a failure of the active insulin receptor kinase domain to couple to yeast (glucose) signalling cascades.

Effect of intravenous dopamine infusion on plasma concentrations of dopamine and dopamine sulfate in men, during and up to 18 h after infusion.

OBJECTIVE: We investigated whether sulfoconjugation contributes to the inactivation of intravenously infused dopamine (DA) in low concentrations with a predominant action on the kidney. METHODS: Plasma DA and dopamine sulfate (DA-S) concentrations were determined during 4 h of intravenous infusion of DA (2 microg/kg/min) and up to 18 h after cessation of infusion. Twenty-seven healthy young subjects participated in the placebo controlled, randomised and double-blind study. RESULTS: Intravenously administered DA was sulfoconjugated rapidly and to a great extent. After starting the infusion, DA levels rose within minutes and reached a steady state after 30-60 min. The steady-state levels averaged 151.3 +/- 8.2 nmol/l. DA-S levels also increased markedly with infusion from 16.7 +/- 9.9 nmol/l at the start of infusion up to 261.2 +/- 24.2 nmol/l at 30 min after cessation of infusion. Plasma DA concentrations after cessation of the infusion decreased rapidly with an initial half-life of elimination of 4.8 min. Concentrations of plasma DA-S declined with a half-life of 4.5 h. Persistent elevations of free and conjugated DA compared with pre-treatment levels were observed even 18 h after cessation. Heart rate and blood pressure remained unchanged both during DA and saline infusion. CONCLUSION: Findings indicate that the sulfoconjugation pathway contributes markedly to the inactivation of intravenously infused DA and seems not to be saturable by DA infusion in low doses.

Three new cytochalasins produced by an endophytic fungus in the genus Rhinocladiella.

Three new cytotoxic cytochalasins (1-3) and the previously reported cytochalasin E (4) were isolated from a culture of the endophytic fungus Rhinocladiella sp. using bioassay-guided fractionation. Extensive NMR and HRCIMS experiments identified these new compounds as 22-oxa-[12]-cytochalasins.

Cryptocin, a potent tetramic acid antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina.

[formula: see text] The endophytic fungus Cryptosporiopsis cf. quercina produces cryptocin in culture. Among other fungi, this unique tetramic acid displays antimycotic activity against Pyricularia oryzae, the causal agent of rice blast disease. Cryptocin also possesses activity against a wide variety of plant pathogenic but not human pathogenic fungi. The fine rhomboid-like crystals of cryptocin allowed structural elucidation by X-ray crystallography. The importance of cryptocin to the symbiotic relationship of C. quercina to its hosts is briefly discussed.

Seimatoantlerium tepuiense gen. nov., a unique epiphytic fungus producing taxol from the Venezuelan Guyana.

Seimatoantlerium gen. nov., type species, S. tepuiense sp. nov. is proposed for an acervular fungus producing 4-septate, holoblastic conidia with 6-8 unbranched, apical appendages that dehisce as an appendage apparatus and also commonly possessing one or two exogenous basal appendages as well as a pedicel. It is compared with Seimatosporium, Seimatosporiopsis, and other genera. It is epiphytic on Maguireothamnus speciosus, a rubiaceous plant endemic to the tepuis of southeastern Venezuela. It produces the anti-oomycetous anticancer compound, taxol, as shown by immunological and spectroscopic methods. Taxol production is discussed relative to the ability of this fungus to exist in an extremely moist ecosystem, as well as to its relationship to other plant associated fungi.

Intravenous nitrates for pharmacological stimulation during head-up tilt testing in patients with suspected vasovagal syncope and healthy controls.

Nitrates may be used for pharmacological stimulation during tilt testing for the diagnosis of vasovagal syncope. In this study we assessed the diagnostic value of intravenous nitrates during tilt testing in patients with a typical history of vasovagal syncope. Twenty patients and 23 controls were tilted at 700 for a maximum duration of 30 minutes. After a 10-minute baseline supine phase, the test started with a continuous nitrate infusion at 1 microg/kg/min and increased every 5 minutes by 1 microg/kg/min, to a maximum of 6 microg/kg/min at the end of the test. The test was ended if the subjects developed a positive response (syncope or presyncope). Nineteen patients (95%) and 17 (74%) of the controls had a positive response. At test end sensitivity was 95%, but specificity was 26% and accuracy was 58%. Receiver operator characteristics (ROC) analysis revealed a maximum accuracy of 79% at 18 minutes, with a sensitivity of 80% and a specificity of 78%. Intravenous nitrates during tilt testing in patients with typical clinical criteria of vasovagal syncope is highly effective in provoking vasovagal syncope. Based on the ROC analysis, a maximum accuracy of 79% was attained at 18 minutes (at a dose of 4 microg/kg/min), suggesting a good diagnostic performance when tilt duration is limited to this point. A positive result requiring more than 18 minutes of stimulated tilting should be interpreted with caution, due to the accompanying considerable decrease of specificity.