Outpatient inotropic therapy in heart transplant candidates: should its use influence waiting list priority status?

We have initiated large-scale sequencing of the third smallest chromosome of the CL Brener strain of Trypanosoma cruzi and we report here the complete sequence of a contig consisting of three cosmids. This contig covers 93.4 kb and has been found to contain 20-30 novel genes and several repeat elements, including a novel chromosome 3-specific 400-bp repeat sequence. The intergenic sequences were found to be rich in di- and trinucleotide repeats of varying lengths and also contained several known T. cruzi repeat elements. The sequence contains 29 open reading frames (ORFs) longer than 700 bp, the longest being 5157 bp, and a large number of shorter ORFs. Of the long ORFs, seven show homology to known genes in parasites and other organisms, whereas four ORFs were confirmed by sequencing of cDNA clones. Two shorter ORFs were confirmed by a database homology and a cDNA clone, respectively, and one RNA gene was identified. The identified genes include two copies of the gene for alanine-aminotransferase as well as genes for glucose-6-phosphate isomerase, protein kinases and phosphatases, and an ATP synthase subunit. An interesting feature of the sequence was that the genes appear to be organized in two long clusters containing multiple genes on the same strand. The two clusters are transcribed in opposite directions and they are separated by an approximately 20-kb long, relatively GC-rich sequence, that contains two large repetitive elements as well as a pseudogene for cruzipain and a gene for U2snRNA. It is likely that this strand switch region contains one or more regulatory and promoter regions. The reported sequence provides the first insight into the genome organization of T. cruzi and shows the potential of this approach for rapid identification of novel genes. [The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AF052831-AF052833.]     

In vitro culture retards spontaneous activation of cell cycle progression and cortical granule exocytosis that normally occur in in vivo unfertilized mouse eggs

A microchip method has been developed for massive and parallel thermodynamic analyses of DNA duplexes. Fluorescently labeled oligonucleotides were hybridized with oligonucleotides immobilized in the 100 x 100 x 20 mum gel pads of the microchips. The equilibrium melting curves for all microchip duplexes were measured in real time in parallel for all microchip duplexes. Thermodynamic data for perfect and mismatched duplexes that were obtained using the microchip method directly correlated with data obtained in solution. Fluorescent labels or longer linkers between the gel and the oligonucleotides appeared to have no significant effect on duplex stability. Extending the immobilized oligonucleotides with a four-base mixture from the 3'-end or one or two universal bases (5-nitroindole) from the 3'- and/or 5'-end increased the stabilities of their duplexes. These extensions were applied to increase the stabilities of the duplexes formed with short oligonucleotides in microchips, to significantly lessen the differences in melting curves of the AT- and GC-rich duplexes, and to improve discrimination of perfect duplexes from those containing poorly recognized terminal mismatches. This study explored a way to increase the efficiency of sequencing by hybridization on oligonucleotide microchips.     

Effects of increased solar ultraviolet radiation on materials

Synthetic polymers such as plastics, as well as naturally occurring polymer materials such as wood, are extensively used in building construction and other outdoor applications where they are routinely exposed to sunlight. The UV-B content in sunlight is well known to affect adversely the mechanical properties of these materials, limiting their useful life. Presently their outdoor lifetimes depend on the use of photostabilizers in the case of plastics and on protective surface coatings in the case of wood. Any increase in the solar UV-B content due to a partial ozone depletion would therefore tend to decrease the outdoor service life of these materials. It is the synergistic effect of increased UV radiation with other factors such as the temperature that would determine the extent of such reduction in service life. The increased cost associated with such a change would be felt unevenly across the globe. Those developing countries that depend on plastics as a prime material of construction and experience high ambient temperatures are likely to be particularly affected in spite of the relatively small fractional decrease in ozone at those locations. Assessment of the damage to materials, associated with ozone depletion, requires a knowledge of the wavelength dependence as well as the dose-response characteristics of the polymer degradation processes of interest. While the recent literature includes some reliable spectral sensitivity data, little dose-response information has been reported, so it is difficult to make such assessments reliably at the present time. This is particularly true for the naturally occurring materials popularly used in construction applications. To maintain polymers at the same useful lifetime in spite of increased solar UV-B content, the amount of photostabilizers used in the formulations might be increased. This strategy assumes that conventional stabilizers will continue to be effective with the spectrally altered UV-B-enhanced solar radiation. While the present understanding of the degradation chemistry suggests the strategy to have merit, its effectiveness, in an altered solar radiation environment, has not been demonstrated for common polymers. The availability of these data is crucial for reliably estimating the cost of mitigating the increased damage to materials as a result of a possible partial depletion of the ozone layer using this approach.     

Relation between chemosensitivity and the ventilatory response to exercise in chronic heart failure

OBJECTIVES: This study sought to establish the chemosensitivity of patients with chronic heart failure. BACKGROUND: The ventilatory response to exercise is often increased in patients with chronic heart failure, as characterized by the steeper regression slope relating minute ventilation to carbon dioxide output. We hypothesized that the sensitivity of chemoreceptors may be reset and may in part mediate the exercise hyperpnea seen in this condition. METHODS: Hypoxic and peripheral hypercapnic chemosensitivity were studied in 38 patients with chronic heart failure (35 men, 3 women; mean [+/-SE] age 60.2 +/- 1.3 years; radionuclide left ventricular ejection fraction 25.7 +/- 2.3%) and 15 healthy control subjects (11 men, 4 women; mean age 54.9 +/- 3.0 years) using transient inhalations of pure nitrogen and single breaths of 13% carbon dioxide, respectively. The change in chemosensitivity during mild exercise (25 W) was assessed in the first 15 patients and all control subjects. Central hypercapnic chemosensitivity was also characterized in 25 patients and 10 control subjects by the rebreathing of 7% carbon dioxide in 93% oxygen. Cardiopulmonary exercise testing was performed in all subjects. RESULTS: Maximal oxygen consumption was 16.6 +/- 0.9 versus 29.7 +/- 2.2 mol/kg per min (p < 0.0001), and the ventilation-carbon dioxide output regression slope was 37.2 +/- 1.5 versus 26.5 +/- 1.4 (p < 0.0001) in patients and control subjects, respectively. Hypoxic and central hypercapnic chemosensitivity were enhanced in patients (0.707 +/- 0.076 vs. 0.293 +/- 0.056 liters/min per % arterial oxygen saturation [SaO2], p = 0.0001 and 3.15 +/- 0.41 vs. 2.02 +/- 0.25 liters/min per mm Hg, p = 0.025, respectively) and correlated significantly with the ventilatory response to exercise. Hypoxic chemosensitivity was augmented during exercise in patients and in control subjects but remained higher in the former (1.530 +/- 0.27 vs. 0.685 +/- 0.12 liters/min per %SaO2, p = 0.01). The peripheral hypercapnic chemosensitivity of patients at rest and during exercise was similar to that in control subjects, consistent with its lesser contribution to overall carbon dioxide chemosensitivity. CONCLUSIONS: Enhanced hypoxic and central hypercapnic chemosensitivity may play a role in mediating the increased ventilatory response to exercise in chronic heart failure.     

Mood stabilizers: shared mechanisms of action at postsynaptic signal-transduction and kindling processes

Several distinct classes of agents have demonstrated efficacy as mood stabilizers in patients with bipolar disorder. It may be reasonable to assume that these agents share one or more common mechanisms of action. This paper will explore the hypothesis that all effective mood stabilizers exert their actions through inhibition of postsynaptic signal-transduction and kindling processes. A literature search was performed for all currently used mood stabilizers to identify reports of mood-stabilizer action in postreceptor cell-signaling and kindling processes. Most effective mood stabilizers appear to inhibit intracellular calcium mobilization through several distinct mechanisms. In addition, several mood stabilizers appear to diminish generation of second-messenger molecules from the membrane phospholipids phosphatidylinositol and phosphatidylcholine, inhibit the activity of protein kinases, and directly inhibit activity of G-proteins. Finally, all established mood stabilizers also exhibit antikindling effects. All of these mechanisms of action could dampen excessive intracellular and intercellular signaling, which may be a core feature of the pathophysiology of bipolar disorder. The observation that all effective mood stabilizers inhibit both kindling and signal-transduction pathways suggests that these processes are intimately linked. We hypothesize that an effective mood stabilizer must possess some specific minimum inhibitory effects at postsynaptic signal-transduction and kindling processes. If this hypothesis is correct, then a rational search for safer and more effective mood-stabilizing agents can begin.     

Rapid ganciclovir susceptibility assay using flow cytometry for human cytomegalovirus clinical isolates

Rapid, quantitative, and objective determination of the susceptibilities of human cytomegalovirus (HCMV) clinical isolates to ganciclovir has been assessed by an assay that uses a fluorochrome-labeled monoclonal antibody to an HCMV immediate-early antigen and flow cytometry. Analysis of the ganciclovir susceptibilities of 25 phenotypically characterized clinical isolates by flow cytometry demonstrated that the 50% inhibitory concentrations (IC50s) of ganciclovir for 19 of the isolates were between 1.14 and 6.66 microM, with a mean of 4.32 microM (+/-1.93) (sensitive; IC50 less than 7 microM), the IC50s for 2 isolates were 8.48 and 9.79 microM (partially resistant), and the IC50s for 4 isolates were greater than 96 microM (resistant). Comparative analysis of the drug susceptibilities of these clinical isolates by the plaque reduction assay gave IC50s of less than 6 microM, with a mean of 2.88 microM (+/-1.40) for the 19 drug-sensitive isolates, IC50s of 6 to 8 microM for the partially resistant isolates, and IC50s of greater than 12 microM for the four resistant clinical isolates. Comparison of the IC50s for the drug-susceptible and partially resistant clinical isolates obtained by the flow cytometry assay with the IC50s obtained by the plaque reduction assay showed an acceptable correlation (r2 = 0.473; P = 0.001), suggesting that the flow cytometry assay could substitute for the more labor-intensive, subjective, and time-consuming plaque reduction assay.     

Athletic amenorrhea and its consequences. Hard physical training at an early age can cause serious bone damage

Strenuous exercise in women is associated with a high incidence of menstrual dysfunction, including amenorrhoea. Athletic amenorrhoea is most common among long-distance runners and ballet dancers, with a prevalence of up to 66 per cent. It is of hypothalamic origin, the pulsatile release of GnRH (gonadotrophin-releasing hormone) being disturbed during exercise, resulting in low gonadotrophin and oestrogen levels. Accumulated evidence suggests athletic amenorrhoea to be related to energy deficiency or to the eating disorders that are prevalent among athletes. The long-term consequences of amenorrhoea are premature osteoporosis and increased risk of musculoskeletal injury. Elite training in young girls tends to delay pubertal development, resulting in decreased bone mass accumulation and reduced growth potential.     

Medical practice guidelines in malpractice litigation: an early retrospective

The role of practice guidelines in malpractice litigation has been discussed in some theoretical detail. Little information is available, however, on how courts use guidelines or on the effort of state legislatures to explicitly link guideline compliance with malpractice defenses. We review all relevant case law and legislative enactments to shed light on the influence of medical malpractice on guidelines. We also use data from a nationwide survey of malpractice attorneys to supplement our legal analysis. Although guidelines are being used for both inculpatory and exculpatory purposes in common-law litigation (a two-way street), legislatures are interested in applying them only for exculpatory purposes (a one-way street).