High-resolution profiles of the Streptococcus mitis CSP signaling pathway reveal core and strain-specific regulated genes.

BACKGROUND: In streptococci of the mitis group, competence for natural transformation is a transient physiological state triggered by competence stimulating peptides (CSPs). Although low transformation yields and the absence of a widespread functional competence system have been reported for Streptococcus mitis, recent studies revealed that, at least for some strains, high efficiencies can be achieved following optimization protocols. To gain a deeper insight into competence in this species, we used RNA-seq, to map the global CSP response of two transformable strains: the type strain NCTC12261T and SK321. RESULTS: All known genes induced by ComE in Streptococcus pneumoniae, including sigX, were upregulated in the two strains. Likewise, all sets of streptococcal SigX core genes involved in extracellular DNA uptake, recombination, and fratricide were upregulated. No significant differences in the set of induced genes were observed when the type strain was grown in rich or semi-defined media. Five upregulated operons unique to S. mitis with a SigX-box in the promoter region were identified, including two specific to SK321, and one specific to NCTC12261T. Two of the strain-specific operons coded for different bacteriocins. Deletion of the unique S. mitis sigX regulated genes had no effect on transformation. CONCLUSIONS: Overall, comparison of the global transcriptome in response to CSP shows the conservation of the ComE and SigX-core regulons in competent S. mitis isolates, as well as species and strain-specific genes. Although some S. mitis exhibit truncations in key competence genes, this study shows that in transformable strains, competence seems to depend on the same core genes previously identified in S. pneumoniae.

Simultaneous repair of two large cranial defects using rapid prototyping and custom computer-designed titanium plates: a case report.

Custom titanium cranioplasty plates, manufactured by a variety of techniques, have been used to repair a range of cranial defects. The authors present a case where two relatively large, adjacent cranial defects were repaired by custom computer-designed titanium plates. The two plates were designed and fabricated simultaneously using a unique methodology. A 28-year-old woman underwent a corpus callosotomy for medically intractable epilepsy. The surgery was complicated by unexpected haemorrhage which necessitated a second craniotomy. Subsequent deep infection required the removal of bilateral bone flaps, presenting a challenge in the reconstruction of extensive, bilateral but asymmetrical cranial defects. The patient underwent a head computed tomography scan, from which a rapid-prototype model of the skull was produced. The surfaces for the missing cranial segments were generated virtually using a combination of software products and two titanium plates that followed these virtual contours were manufactured to cover the defects. The cranioplasty procedure to implant both titanium cranial plates was performed efficiently with no intra-operative complications. Intra-operatively, an excellent fit was achieved. The careful planning of the plates enhanced the relative ease with which the cranial defects were repaired with an excellent cosmetic outcome.

Morphology and related chemistry of small lunar particles from tranquillity base.

Glass spherules show multiple high-velocity impact craters and are coated with small particles including glass, plagioclase, clinopyroxene, ilmenite, olivine, chromite, rock fragments, and frozen droplets of iron, nickel-iron, and troilite. These spherules passed through an impact cloud of hot fragmental material, condensing iron-rich vapor and high-velocity projectiles. Breccia contains concentric, accretionary lapilli units and appears to be a sintered deposit from a hot lunar base surge generated by impact.

Complete genome sequence of a virulent isolate of Streptococcus pneumoniae.

The 2,160,837-base pair genome sequence of an isolate of Streptococcus pneumoniae, a Gram-positive pathogen that causes pneumonia, bacteremia, meningitis, and otitis media, contains 2236 predicted coding regions; of these, 1440 (64%) were assigned a biological role. Approximately 5% of the genome is composed of insertion sequences that may contribute to genome rearrangements through uptake of foreign DNA. Extracellular enzyme systems for the metabolism of polysaccharides and hexosamines provide a substantial source of carbon and nitrogen for S. pneumoniae and also damage host tissues and facilitate colonization. A motif identified within the signal peptide of proteins is potentially involved in targeting these proteins to the cell surface of low-guanine/cytosine (GC) Gram-positive species. Several surface-exposed proteins that may serve as potential vaccine candidates were identified. Comparative genome hybridization with DNA arrays revealed strain differences in S. pneumoniae that could contribute to differences in virulence and antigenicity.

Complete Genome Sequence of Streptococcus pneumoniae Serotype 19F Strain EF3030.

We report the complete genome sequence of Streptococcus pneumoniae EF3030, a serotype 19F isolate that colonizes the nasopharynx of mice while being mostly noninvasive. Such attributes make this strain highly attractive in pneumococcal carriage studies. The availability of its complete genomic sequence is likely to advance studies in the field.

Characterization of bovine lymphocytes stimulated in vitro by Dictyocaulus viviparus homogenate.

Adult Dictyocaulus viviparus homogenate induced proliferation of lymphocytes from naïve cattle. We characterized the responding cells by carboxyfluorescein diacetate succinimidyl ester (CFSE) loading, for detection of proliferation, and antibody labelling for cell surface molecules. Lymphocytes expressing CD4, CD8 and gamma/delta TCR, rather than Ig expressing cells, proliferated after in vitro stimulation with D. viviparus homogenate. Of gamma/delta TCR expressing cells, both CD8, WC1.1 and WC1.2 co-expressing cells proliferated. Moreover, gamma/delta T cells expressing MHC class II proliferated to a higher extent than those negative for MHC class II. Of CD4 and CD8 expressing lymphocytes, both those co-expressing CD45R and CD45R0 proliferated. Among CD4 expressing lymphocytes, those that were CD45R0 positive had a larger proportion of proliferated cells than did CD45R positive cells. Compared to stimulation with Con A, the proportion of dividing cells after D. viviparus stimulation was smaller although the cells had divided more times. Furthermore, we also compared in vitro responses of peripheral blood mononuclear cells collected before and after two subsequent infections with D. viviparus, but no clear acquired responses could be detected. Overall, this suggests that most T lymphocytes are stimulated by the D. viviparus homogenate rather than any particular lymphocyte subpopulation.

Prospects for elucidating the phylogeny of the Apicomplexa.

The Apicomplexa is traditionally considered to contain four clearly defined groups, but the phylogenetic relationships among these groups, and their subsequent taxonomic arrangement, are currently unclear. Here, I consider five of the important criteria that need to be met before radical taxonomic changes are made, in relation to phylogenetic analyses. Unfortunately, at least four of these criteria indicate that the prospects for elucidating the phylogeny and taxonomy of the Apicomplexa are not good in the short term. A lot more pertinent data need to be collected, both in terms of taxa sampled and genes sequenced, before any concordance among phylogenies is likely to emerge; and someone (e.g. a consortium) needs to coordinate the active collection and analysis of these data if any worthwhile progress is to be made.

Emergency percutaneous coronary intervention (PCI) for the care of patients with ST-elevation myocardial infarction (STEMI).

There is general consensus that emergency percutaneous coronary intervention (PCI) is the preferred treatment for patients with ST-elevation myocardial infarction (STEMI), so long as it can be delivered in a timely fashion, by an experienced' operator and cardiac catheterization laboratory (CCL) team. STEMI is both a functional and structural issue. Although it has been recognized since the work of pioneering cardiologists and surgeons in Spokane, Washington, that approximately 88% of patients presenting within 6 hours of onset of STEMI have an occluded coronary artery, it is the pathophysiology of myocardial necrosis, and the varied consequences of necrosis that characterize STEMI. Accordingly, experience' of both primary operator and cardiac catheterization laboratory (CCL) crew, in performing an emergency PCI for STEMI, are as much a function of experience with the treatment of complex MI patients, as experience with coronary intervention. Rapidly achieving normal coronary artery flow, at both the macro and micro vascular levels, is the recognized key to aborting the otherwise progressive wavefront' of myocardial necrosis. The time urgency of decisions (Time is muscle') make emergency PCI for patients with on-going necrosis, more like emergency room (ER) care, than like most in-hospital or outpatient care. In general, most patients with acute coronary syndromes (ACS) are currently thought to have plaque rupture and/or erosion with subsequent thrombosis and embolization. Consequences of thrombo-embolism, such as slow flow' or no-reflow' are in addition to, the structural (anatomic) considerations of PCI in stable patients (such as ostial location; bifurcation involvement; heavy calcification; tortuosity of lesion or access to it; length of disease; caliber of infarct-artery; etc.). Good quality studies have provided strong support for the specific added value of glycoprotein IIb/IIIa inhibitors (especially abciximab), dual antiplatelet therapy (the addition of the thienopyridine, clopidogrel, to aspirin use), and bare-metal stents (BMS), for a broad range of STEMI patients. The added value of drug-eluting stents (DES) to bare-metal stents (BMS), primarily in terms of reducing restenosis and repeat revascularization, is supported by several randomized trials, and a number of registries, despite its being off-label' from a regulatory standpoint. The recognition of late stent thrombosis (LST) has raised additional issues, in choosing between these two options for specific STEMI patients. The added value of a number of other mechanical approaches to coronary thrombus, such as thrombus removal devices, and/or distal protection, are more controversial, and perhaps, patient specific. Whether intravascular ultrasound guidance (IVUS) for stent use should be used for the majority, or even a specific minority, of STEMI patients, is also controversial; late-stent thrombosis provides a counter-point. The advantages of developing a network approach to STEMI care, so as to optimize the number of patients receiving timely reperfusion, have been demonstrated in Prague, Denmark, and Minneapolis, among many places. The benefits of both bivalirudin (anti-thrombin drug with efficacy against clot-bound thrombin, which does not appear to stimulate platelets) and abciximab (glycoprotein IIb/IIIa inhibitor which is antibody to platelet receptors), as PCI adjuncts generally, and for STEMI patients, in particular, are supported by multiple trials. The specific choice of administering the bolus dose of either, or both, drugs via intra-coronary (IC) injection follows the precedents' of IC thrombolytics, and IC small-vessel vasodilators for no-reflow', but it has not been tested by prospective, randomized trials. Although rapid reperfusion is the first objective, one cannot ignore the other components of the oxygen delivery chain, and the importance of each of these components to on-going delivery of oxygen to all vital organs. A balance must be struck between doing those control' things which serve to stabilize other vital components of the oxygen-delivery chain, without digressing too long from the job of re-establishing brisk coronary flow. The clinical and angiographic heterogeneity of STEMI patients and the array of available therapeutic approaches make it impossible to obtain specific randomized trial direction for many of the clinical decisions in an individual emergency PCI for STEMI. There are a range of reasonable/ appropriate therapeutic choices for a given emergent PCI performed by multiple experienced and competent operators. The treatment of STEMI, and high-risk non-STEMI, patients, by means of emergent PCI, is among the most challenging and rewarding arenas in contemporary medicine.

Biological control of sheep parasites using Duddingtonia flagrans: trials on commercial farms in Sweden.

Trials were conducted on 3 commercial sheep farms in Sweden to assess the effect of administering spores of the nematode trapping fungus, Duddingtonia flagrans, together with supplementary feed to lactating ewes for the first 6 weeks from turn-out on pastures in spring. Also control groups of ewes, receiving only feed supplement, were established on all 3 farms. Groups were monitored by intensive parasitological investigation. The ewes and their lambs were moved in late June to saved pastures for summer grazing, the lambs receiving an anthelmintic treatment at this time. After approximately 6 weeks on summer pasture the lambs were weaned, treated a second time with anthelmintic, and returned to their original lambing pastures for finishing. Decisions as to when lambs were to be marketed were entirely at the discretion of the farmer co-operators. No difference in lamb performance was found between the two treatments on all three farms. This was attributed to the high levels of nutrition initially of the ewes limiting their post-partum rise in nematode faecal egg counts in spring, which in turn resulted in low levels of nematode infection on pastures throughout the autumn period. Additionally, pastures were of good quality for the lambs during the finishing period, so they grew at optimal rates as far as the farmers were concerned.

Comprehensive Transcriptome Profiles of Streptococcus mutans UA159 Map Core Streptococcal Competence Genes.

In Streptococcus mutans, an oral colonizer associated with dental caries, development of competence for natural genetic transformation is triggered by either of two types of peptide pheromones, competence-stimulating peptides (CSPs) (18 amino acids [aa]) or SigX-inducing peptides (XIPs) (7 aa). Competence induced by CSP is a late response to the pheromone that requires the response regulator ComE and the XIP-encoding gene comS. XIP binds to ComR to allow expression of the alternative sigma factor SigX and the effector genes it controls. While these regulatory links are established, the precise set of effectors controlled by each regulator is poorly defined. To improve the definition of all three regulons, we used a high-resolution tiling array to map global changes in gene expression in the early and late phases of the CSP response. The early phase of the CSP response was limited to increased gene expression at four loci associated with bacteriocin production and immunity. In the late phase, upregulated regions expanded to a total of 29 loci, including comS and genes required for DNA uptake and recombination. The results indicate that the entire late response to CSP depends on the expression of comS and that the immediate transcriptional response to CSP, mediated by ComE, is restricted to just four bacteriocin-related loci. Comparison of the new data with published transcriptome data permitted the identification of all of the operons in each regulon: 4 for ComE, 2 for ComR, and 21 for SigX. Finally, a core set of 27 panstreptococcal competence genes was identified within the SigX regulon by comparison of transcriptome data from diverse streptococcal species. IMPORTANCES. mutans has the hard surfaces of the oral cavity as its natural habitat, where it depends on its ability to form biofilms in order to survive. The comprehensive identification of S. mutans regulons activated in response to peptide pheromones provides an important basis for understanding how S. mutans can transition from individual to social behavior. Our study placed 27 of the 29 transcripts activated during competence within three major regulons and revealed a core set of 27 panstreptococcal competence-activated genes within the SigX regulon.

Coronary angioplasty for elderly patients with "high risk" unstable angina: short-term outcomes and long-term survival.

OBJECTIVES: We sought to compare the short- and long-term mortality rates in patients > or = 70 years old with unstable angina undergoing percutaneous transluminal coronary angioplasty (PTCA) with predicted coronary artery bypass graft surgery (CABG) short-term and U.S. census long-term mortality rates. BACKGROUND: Coronary angioplasty is an alternative revascularization strategy for patients with medically refractory rest angina and a high risk of adverse outcomes with CABG. Patients > or = 70 years old are a specific high risk subset. METHODS: A total of 131 consecutive patients aged > or = 70 years with unstable angina underwent PTCA; 82 (62%) of 131 had been refused CABG. Mortality over time was obtained from the Veterans Affairs Beneficiary Index Records Locator Subsystem. Predicted 30-day CABG-associated mortality was obtained from the Veterans Affairs Cardiac Risk Assessment Model. Mortality over time was expressed with Kaplan-Meier curves. RESULTS: The observed 30-day angioplasty survival rate was 87% compared with the predicted surgical 30-day survival rate of 85.5%. In those patients who survived 6 months after angioplasty (84%), their subsequent 1-, 2-, 3-, 4- and 5-year survival rates were comparable to age-matched subjects in the U.S. census. Mortality in certain subsets known to be at very high risk for CABG-for example, patients who had a previous CABG-was not high in this cohort of elderly subjects. The extremely high risk subsets identified in this PTCA cohort (shock, heart failure, pressors required, balloon pump required) were relatively infrequent subsets. CONCLUSIONS: For selected elderly patients with unstable angina deemed to be at "high risk" or even "prohibitive risk" for CABG, PTCA is an alternative revascularization strategy. The long-term mortality of successfully treated elderly patients is comparable to age-matched subjects. A prospective, multicenter, randomized trial of CABG versus PTCA, which includes patients > or = 70 years old, is being conducted (Veterans Affairs Cooperative Study 385: AWESOME).

Right ventricular function at rest and during exercise in aortic and mitral valve disease.

Right ventricular function at rest and during exercise was examined in a group of patients with symptomatic aortic or mitral valve disease, or both. The right ventricular ejection fraction was less than 45% in 22 of 36 patients at rest and in 12 of 17 subjects at symptom-limited, supine bicycle exercise. The right ventricular ejection fraction failed to increase more than 5% with exercise in 17 of 17 patients. The central venous pressure was greater than 5 mm Hg in 18 of 36 patients at rest and in 13 of 17 patients at maximal exercise. There was a significant inverse relation between rest right ventricular ejection fraction and mean pulmonary artery pressure (r = -0.47, p less than 0.05) and between rest right ventricular ejection fraction and mean central venous pressure (r = -0.39, p less than 0.05). There was no significant relation between the exercise values of these variables. In individual patients, the changes in right ventricular ejection fraction and pulmonary artery pressure with graded exercise were nonlinear. It is concluded that right ventricular function is not a simple function of pulmonary artery pressure at rest or during exercise in aortic and mitral valve disease. Less than one-quarter of the variation in right ventricular ejection fraction at rest can be explained by the variation in pulmonary artery pressure, and the finding of a normal (greater than 45%) right ventricular ejection fraction does not reliably exclude the possibility of pulmonary hypertension in a patient with valvular heart disease.

Percutaneous transluminal angioplasty of saphenous vein grafts for medically refractory unstable angina.

OBJECTIVES: We attempted to answer the question, Is balloon angioplasty a reasonable alternative to repeat coronary artery bypass graft surgery in patients with previous coronary bypass graft surgery, medically refractory unstable angina and vein graft lesions? BACKGROUND: Patients with medically refractory unstable angina need revascularization. Patients with previous coronary artery bypass graft surgery and medically refractory angina are at "high risk" for adverse outcomes with repeat coronary bypass graft surgery. Conversely, patients with angioplasty of old vein grafts are also at "high risk" for adverse outcomes. METHODS: Balloon angioplasty of 89 lesions in saphenous vein grafts was performed in 75 consecutive patients with medically refractory unstable angina. Of these 75 patients, 24 (32%) had myocardial infarct within 30 days, 23 (31%) had left ventricular ejection fraction < 0.35, and 50 (67%) had major comorbidity. Patients underwent standard balloon angioplasty with aggressive use of intravenous and intracoronary heparin, urokinase, nitroglycerin, oral aspirin, calcium channel blocking agents and coumadin. RESULTS: Angiographic success (reduction of stenosis < or = 50% without major complication) was seen in 84 of 89 lesions. Clinical success (angiographic success plus hospital discharge without major complication) was seen in 70 of 75 patients. During index hospitalization, two patients (3%) died, two (3%) had nonfatal infarcts, and one (1%) had emergency reoperation (coronary bypass graft surgery). In late follow up (3 to 66 months), 14 (20%) patients were lost to follow-up, 17 (23%) had repeat percutaneous transluminal coronary angioplasty, 2 (3%) had late bypass graft reoperation, 18 (25%) had late death, and 1 (< 1%) had a heart transplant. Of the 41 patients alive after one or more angioplasties, 25 have little or no angina, and 16 have occasional or more angina. We compared long-term survival rate in these 75 patients with a cohort of patients with high risk, unstable angina from the Veterans Affairs Surgical Registry (2,570 patients). The 30-day survival rate was better in patients with coronary angioplasty (97% vs. 92%, p < 0.05), but by 6 months there was no difference, and by 5 years a trend toward a higher survival rate with coronary artery bypass graft surgery was seen. CONCLUSIONS: Balloon angioplasty of saphenous vein grafts with aggressive adjunctive pharmacotherapy is a reasonable alternative to repeat coronary bypass graft surgery in patients with medically refractory unstable angina, previous coronary bypass graft surgery and saphenous vein narrowing.

Right ventricular dysfunction and the exercise limitation of chronic obstructive pulmonary disease.

This study examined right ventricular function during exercise in patients with chronic obstructive pulmonary disease to answer the following questions: Is there a significant correlation between oxygen consumption at maximal exercise and exercise right ventricular ejection fraction? Does the right ventricular ejection fraction response to exercise correlate with exercise changes in pulmonary artery pressure, total pulmonary resistance or pulmonary vascular resistance? Which combinations of cardiac, ventilatory and blood gas variables are the best predictors of oxygen consumption at maximal exercise? Twenty-six patients with stable chronic obstructive pulmonary disease performed symptom-limited supine bicycle exercise with simultaneous hemodynamic and radionuclide ventriculographic measurements. The oxygen consumption at maximal exercise correlated with the exercise right ventricular ejection fraction (n = 21, r = 0.66; p less than 0.005), exercise stroke volume (r = 0.68; p less than 0.001), exercise cardiac output (r = 0.77; p less than 0.00005) and exercise ventilation (r = 0.85; p less than 0.00001). The change in right ventricular ejection fraction from rest to exercise correlated inversely with the change from rest to exercise in total pulmonary resistance (r = -0.51; p less than 0.05) but not with the change in mean pulmonary pressure (r = -0.37) or in pulmonary vascular resistance (r = 0.09). Multivariate analysis showed that the variables giving the highest combined correlation with oxygen consumption were ventilation and right ventricular ejection fraction (r = 0.95, adjusted r2 = 0.88). These results suggest that exercise oxygen consumption of patients with chronic obstructive pulmonary disease is related to right ventricular systolic function, exercise right ventricular dysfunction is related, in part, to abnormal exercise total pulmonary resistance, and exercise limitation in chronic obstructive pulmonary disease occurs as a result of the dynamic interaction between disordered right heart function and ventilation.

Percutaneous coronary intervention versus coronary artery bypass graft surgery for patients with medically refractory myocardial ischemia and risk factors for adverse outcomes with bypass: a multicenter, randomized trial. Investigators of the Department of Veterans Affairs Cooperative Study #385, the Angina With Extremely Serious Operative Mortality Evaluation (AWESOME).

BACKGROUND: Percutaneous coronary intervention (PCI) and coronary artery bypass graft surgery (CABG) are being applied to high-risk populations, but previous randomized trials comparing revascularization methods have excluded a number of important high-risk groups. OBJECTIVES: This five-year, multicenter, randomized clinical trial was designed to compare long-term survival among patients with medically refractory myocardial ischemia and a high risk of adverse outcomes assigned to either a CABG or a PCI strategy, which could include stents. METHODS: Patients from 16 Veterans Affairs Medical Centers were screened to identify myocardial ischemia refractory to medical management and the presence of one or more risk factors for adverse outcome with CABG, including prior open-heart surgery, age >70 years, left ventricular ejection fraction <0.35, myocardial infarction within seven days or intraaortic balloon pump required. Clinically eligible patients (n = 2,431) underwent coronary angiography; 781 were angiographically acceptable; 454 (58% of eligible) patients consented to random assignment between CABG and PCI. RESULTS: A total of 232 patients was randomized to CABG and 222 to PCI. The 30-day survivals for CABG and PCI were 95% and 97%, respectively. Survival rates for CABG and PCI were 90% versus 94% at six months and 79% versus 80% at 36 months (log-rank test, p = 0.46). CONCLUSIONS: Percutaneous coronary intervention is an alternative to CABG for patients with medically refractory myocardial ischemia and a high risk of adverse outcomes with CABG.

How old are the extant lineages of Toxoplasma gondii?

Most known isolates of Toxoplasma gondii belong to one of only three lineages, which are presumed to be clonal. Three models have been proposed for the evolutionary relationship of these lineages to the other extant lineages: Model (a) proposing that all lineages are derived from a most recent common ancestor (MRCA) in the distant past, Model (b) that all lineages are derived from a MRCA in the very recent past, and Model (c) that the clonal lineages share a recent MRCA but are related to the other lineages only in the distant past. Here, I test these models using DNA intron and coding-sequence data for loci at 14 genes, using three different methods to calculate the time of the MRCA. All of the calculations agree that the MRCA of the clonal lineages was > 70% of the age of the MRCA of all lineages, thus favouring Model (a). The MRCA may have existed approximately 150,000 years ago, with the clonal lineages expanding in prevalence approximately 10,000 years ago.

Multivessel percutaneous coronary intervention: a new paradigm for a new century.

Percutaneous coronary intervention (PCI) was introduced in 1977, to treat single, discrete lesions in patients with stable symptoms, and favorable anatomy. Increased operator experience and numerous technical advances, notably including bare-metal stents (BMS) and then drug-eluting stents (DES), and a number of adjunctive pharmacologic modalities, have allowed for the gradual application of PCI to far more diverse clinical and anatomic subsets. The proper role of PCI, compared with both medical therapy and coronary artery bypass graft (CABG) surgery, is evolving. This manuscript is synthesized from literature review of randomized clinical trials, and some cohort investigations, and 21 years of PCI experience, which has been focused on high-risk patients. Current American College of Cardiology/ American Heart Association/ Society for Cardiac Angiography and Interven-tions (ACC/AHA/ SCA&I) Guidelines, and most research, are based upon a perspective (conventional paradigm') that categorizes patients primarily based upon number of major' coronary (left anterior descending artery, circumflex and right) arteries with a >70% stenosis, and whether left ventricular function is normal or abnormal (left ventricular ejection fraction < or >0.50). This paradigm developed when: a) CABG was the only revascularization option, and b) medical therapy was quite limited. All of the trials demonstrating survival benefit with aspirin, clopidogrel, statins, angiotensin-converting-enzyme inhibition (ACE-I) or angiotensin receptor blockers (ARB), and beta-blockers; coupled with the PCI versus thrombolytics trials in ST-elevation myocardial infarction (STEMI), and non-STEMI strategy trials, both of which include revascularization almost exclusively by PCI, make this conventional paradigm, outdated and counterproductive. Attempts to compare CABG and PCI have de-emphasized their very different advantages and disadvantages. The new paradigm makes major division of patients based upon whether patients have stable or unstable: a) ischemia and b) hemodynamics, and c) whether they are having acute myocardial infarction (MI). The first issue to be settled is whether the patient is likely to benefit from revascularization. If little or no benefit can be envisioned, the patient should be managed medically. Unstable ischemia and unstable hemodynamics, and acute MI, all favor emergent or urgent revascularization. Clinical features, which generally favor PCI for revascularization, include hemodynamic instability, STEMI and non-STEMI; and severe comorbidity, particularly cerebral, pulmonary, or hepatic comorbidity. Anatomic features, which generally favor CABG, include unprotected left main stenosis, especially involving the bifurcation, one or more graftable chronic total occlusions (CTO), and bifurcation disease with large important side-branches. Old age and severely reduced left ventricular function are associated with higher risks, with either CABG or PCI. Small caliber and diffusely diseased vessels may imply lower expectation of success, by either CABG or PCI.

Changes in right ventricular relaxation during acute anterior myocardial infarction in pigs.

To determine the effect of an anteroseptal myocardial infarction on right ventricular systolic and diastolic function, we studied 12 pigs before and 1 h after left anterior descending coronary artery occlusion. Total arterial occlusion was achieved by the percutaneous, transcatheter placement of a 1 mm Teflon plug into the mid portion of the artery. The resulting infarction involved 28 (SEM 3)% of the left ventricular wall, in the anterior and septal regions. A small rim of the right ventricular free wall adjacent to the septum and the right ventricular apex were also affected. End diastolic pressures in both ventricles rose significantly: left ventricular from 12(1) to 20(2) mm Hg and right ventricular from 8(1) to 10(1) mm Hg. Right ventricular peak systolic pressure increased from 29(2) to 35(2) mm Hg while left ventricular peak systolic pressure did not change. One hour after infarction the half time of isovolumic relaxation of the right ventricle was prolonged from 6.9(0.5) to 8.7(0.4) ms. Ejection fraction in both ventricles was depressed: from 46(1) to 34(2)% in the right ventricle and from 69(3) to 49(3) in the left ventricle. There was no change in either right or left ventricular dP/dt. These data suggest that right ventricular systolic and diastolic dysfunction occurs as the result of an anteroseptal myocardial infarction in pigs.

Genome editing by natural genetic transformation in Streptococcus mutans.

Classical mutagenesis strategies using selective markers linked to designed mutations are powerful and widely applicable tools for targeted mutagenesis via natural genetic transformation in bacteria and archaea. However, the markers that confer power are also potentially problematic as they can be cumbersome, risk phenotypic effects of the inserted genes, and accumulate as unwanted genes during successive mutagenesis cycles. Alternative mutagenesis strategies use temporary plasmid or cassette insertions and can in principle achieve equally flexible mutation designs, but design of suitable counter-selected markers can be complex. All these drawbacks are eased by use of direct genome editing. Here we describe a strategy for directly editing the genome of S. mutans, which is applied to the widely studied reference strain UA159 (ATCC 700610) and has the advantage of extreme simplicity, requiring construction of only one synthetic donor amplicon and a single transformation step, followed by a simple PCR screen among a few dozen clones to identify the desired mutant. The donor amplicon carries the mutant sequence and extensive flanking segments of homology, which ensure efficient and precise integration by the recombination machinery specific to competent cells. The recipients are highly competent cells, in a state achieved by treatment with a synthetic competence pheromone.

Construction and properties of a new insertion vector, pJDC9, that is protected by transcriptional terminators and useful for cloning of DNA from Streptococcus pneumoniae.

A new Escherichia coli plasmid cloning vector, pJDC9, was constructed by replacing the TcR determinant of pMB9 with the erythromycin-resistance ermB determinant and the lacZ alpha gene of pUC19. Efficient transcriptional terminator signals were positioned at both ends of lacZ alpha. Evidence is presented that protection of the vector by terminator signals enabled cloning of many fragments of DNA from Streptococcus pneumoniae that were unstable in vectors lacking such protection, including pBR322. At the pneumococcal mal locus, three promoter sites required such protection, while overexpression of the malX protein appeared to be lethal despite such protection.