Temporal changes in 12-HETE formation in two models of canine myocardial infarction

Arachidonic acid (AA) metabolism by infarcted canine myocardium was studied and correlated with matched histologic analyses following permanently occluded or reperfused infarction. Histologic analysis of tissues from reperfused infarcts showed a marked acceleration of inflammatory cell invasion and of granulation tissue formation when compared to the occlusive infarct. In the reperfused infarct, polymorphonuclear leukocytes (PMNs) were very prominent at one day after infarction while in the occlusive infarcts the neutrophilic invasion was less intense but more sustained. At one day following reperfused infarction the major arachidonate product, which co-migrated by thin layer chromatography with the mono-hydroxyeicosatetraenoic acids (HETEs), was significantly elevated (254 ± 49 pmoles/gm wet weight, n=3) when compared to normal tissue (48 ± 6 pmoles/gm n=19). This occured at a time when the number of PMNs was maximal in the infarcted tissue. Addition of the calcium ionophore A₂₃₁₈₇ caused a further marked stimulation in HETE production in the one day reperfused infarct but not at the other time points studied. The production of HETE was not significantly different in the infarcted tissue than in the normal tissue at three and seven days following reperfused infarction or at one, three, or seven days after occlusive infarction. The identity of this HETE product was investigated using reverse phase high performance liquid chromatography (RP-HPLC) and gas chromatography-mass spectrometry (GC-MS) and found to be predominantly 12 - hydroxy - 5,8,10,14 - eicosatetraenoic acid (12-HETE) with a small amount of 15-HETE. Thus the production of 12-HETE parallels the number of neutrophils invading the infarcted area of the heart.     

Temporal changes in the outcomes of acute myocardial infarction in Ontario, 1992-1996

BACKGROUND: There is relatively little information available on recent population-based trends in the outcomes of patients who have had an acute myocardial infarction (AMI). We, therefore, conducted a study of temporal trends in the outcomes of AMI patients in Ontario, Canada, between the 1992 and 1996 fiscal years. METHODS: 114,618 AMI patients were discharged from hospitals in Ontario between Apr. 1, 1992, and Mar. 31, 1997. After specific exclusion criteria were applied the final sample of 89,456 patients was divided into 5 cohorts according to the fiscal year of discharge. As part of the Ontario Myocardial Infarction Database project the linked administrative data pertaining to these patients were used to examine cohort characteristics, cardiac procedures used and mortality rates for each of the 5 cohorts over time. RESULTS: There was a significant increase in the percentage of patients in Ontario receiving coronary angiography, percutaneous transluminal coronary angioplasty and coronary artery bypass grafting surgery (p < 0.001) after an AMI between 1992 and 1996. In addition, the overall 30-day risk-adjusted mortality rate declined from 15.5% in 1992 to 14.0% in 1996 (p = 0.001) and the 1-year risk-adjusted mortality rate declined from 23.7% in 1992 to 22.3% in 1996 (p = 0.017). Virtually all of the improvement occurred within 30 days of admission. The absolute decline in 1-year mortality rates was significant for patients under the age of 65 (2.3%, 95% confidence interval [CI] 1.4% to 3.2%) and for males (1.2%, 95% CI 0.2% to 2.2%); absolute declines were not significant for patients 65 years of age or older (0.7%, 95% CI -0.6% to 2.0%) and for female patients (-0.1%, 95% CI -1.7% to 1.5%). Interestingly, post-infarction coronary angiography and coronary artery bypass grafting rates were consistently lower in the older and the female patients throughout the study period. INTERPRETATION: There was a modest improvement in the short- and long-term survival of patients in Ontario after an AMI between 1992 and 1996. The Ontario experience suggests that recent advances in AMI management have been of more benefit to younger and male AMI patients.     

Brain 12-HETE formation in different species,brain regions,and in brain microvessels

We used gas chromatography/mass spectrometry to measure brain 12-HETE (12-Hydroxy-5,8,10,14-eicosatetraenoic acid) formation from endogenous arachidonic acid in different species and different brain regions and in isolated brain microvessels. When blood-free brain slices were incubated for 20 minutes we found that the rabbit and cat brain incubates contained little 12-HETE when compared to rat and mouse brain incubates. Further in vitro studies of various rat brain regions showed a generally even distribution of 12-HETE. When isolated rat or rabbit microvessels were incubated and analyzed, we found 1 and 0.25 g, respectively, of 12-HETE/mg of microvessel protein. Also, rabbit brain had limited or no capacity to actively metabolize tritiated 12-HETE. In summary, these studies show substantial species variation with respect to brain formation of 12-HETE and indicate that the vasculature is a potentially significant contributor to the 12-HETE found in whole brain tissue.     

Temporal changes in matrix metalloproteinase expression and inflammatory response associated with cardiac rupture after myocardial infarction in mice

We previously found that male mice with myocardial infarction (MI) had a high rate of cardiac rupture, which generally occurred at 3 to 5 days after MI. Since matrix metalloproteinases (MMPs) play an important role in infarct healing, tissue repair and extracellular matrix (ECM) remodeling post-MI, we studied the temporal relationship of MMP expression and inflammatory response to cardiac rupture after acute MI. Male C57BL/6J mice were subjected to MI (induced by ligating the left anterior descending coronary artery) and killed 1, 2, 4, 7 or 14 days after MI. MMP-2 and MMP-9 activity in the heart were measured by zymography. Collagen content was measured by hydroxyproline assay. We found that after MI, MMP-9 activity increased as early as 1 day and reached a maximum by 2-4 days, associated with a similar increase in neutrophil and macrophage infiltration in the infarct area. MMP-2 started to increase rapidly within 4 days, reaching a maximum by 7 days and remaining high even at 14 days. Intense macrophage infiltration appeared by 4 days after MI and then gradually decreased within 7 to 14 days. Collagen content was unchanged until 4 days after MI, at which point it increased and remained high thereafter. Our data suggest that in mice, overexpression of MMP-2 and MMP-9 (possibly expressed mainly by neutrophils and macrophages) may lead to excessive ECM degradation in the early phase of MI, impairing infarct healing and aggravating early remodeling which in turn causes cardiac rupture.     

Protein kinase C expression and subcellular distribution in chronic myocardial ischemia: Comparison of two different canine models

We studied protein kinase C (PKC) isozyme expression and activity distribution in two models of chronically ischemic canine myocardium: (1) single vessel obstruction (SVO), produced by tight stenosis of LAD followed by preconditioning and acute ischemia (40 min); (2) three vessel obstruction (3VO), produced by LAD-stenosis and gradual occlusion of right coronary artery and left circumflex. In both models after 8 weeks of chronic ischemia the dogs were either sacrificed or had PTCA of the LAD with a follow up of another 4 weeks. Control dogs were sham operated. PKC activity was measured in subcellular fractions of tissue samples from anterior and posterior regions in the presence of histone and -[³²P]-ATP. PKC isozymes were detected by Western blotting. All regions perfused by the obstructed coronaries were dysfunctional at 8 weeks when compared to baseline, with improvement of anterior wall function after PTCA of LAD. PKC activity was elevated in the membrane fraction of SVO, but unchanged in the 3VO model. PKCs , , and prevailed in cytosol fraction of the controls (cytosol/membrane ratios were ± 3.34, 1.38 and 4.56 for , and , respectively), consistent with PKC activity distribution, while was not detected. There was no significant difference between the groups concerning the relative membrane amount of the isozymes. PKCs and were decreased in the cytosol fraction of both models at 8 weeks (for anterior region, by 56 and 57% in SVO and by 49 and 46% in 3VO, respectively) without there being any differences between anterior and posterior regions, and were low also in the PTCA group. PKC distribution however varied between the two models. The amount of PKC isozyme was downregulated by 45% after 8 weeks of chronic ischemia and returned towards the control values after PTCA in the anterior region of SVO, while it did not change in anterior wall after 8 weeks in 3VO but was significantly decreased (by 47%) in posterior region after PTCA. In conclusion, our results suggest modified PKC signalling in chronically ischemic canine myocardium.     

Sex-related changes in cardiac function following myocardial infarction in mice

Recent awareness of cardiovascular diseases as a number one killer of the middle-aged women has prompted interest in sex differences leading to heart failure (HF). Therefore, we evaluated cardiac function in female and male mice following myocardial infarction (MI) using the Millar pressure-volume (P-V) conductance system in vivo, at time points corresponding to early (2 wk), late compensatory hypertrophy (4 wk), and decompensation (10 wk) to HF. A significant deterioration of the load dependent and independent hemodynamic measurements occurred in both female and male mice during the early phase of hypertrophy. Later, compensatory hypertrophy was marked by a normalization of volumes to control levels in females compared with males. The most notable differences between sexes occurred in the measurements of cardiac contractility during the decompensation to HF. In females, there was a significant improvement in contractility compared with males, which was apparent in the load-independent measurements of preload recruitable stroke work (10 wk post-MI, female=48.7+/-8.0 vs. male=25.2+/-1.8 mmHg, P<0.05) and maximum dP/dt vs. maximum end-diastolic volume (10 wk post-MI, female=359+/-58 vs. male=149+/-28 mmHg.s(-1).microl(-1), P<0.05). Despite these differences, there were no differences in the heart weight to body weight ratio and infarct size between the sexes. These data demonstrate that compensatory hypertrophy is associated with an improvement in contractility and a delayed decompensation to HF in females. However, compensatory hypertrophy in males appears to be undermined by a steady decline in contractility associated with decompensation to HF.     

Spatiotemporal dynamics of reentrant ventricular tachycardias in canine myocardial infarction: pharmacological modulation

During the transition from a slow to rapid depolarization rhythm, rate-dependent sodium channel blockade develops progressively and increases from beat to beat under procainamide but more abruptly under lidocaine. We investigated the consequences of such differences on the dynamic course and stability of reentrant tachycardias at their onset. Procainamide and lidocaine were infused to equipotent plasma concentrations in canines with three-day-old myocardial infarction. We measured the activation times (ms) and maximum slopes of negative deflections in activation complexes (absolute value: /-dV/dt(max)/ in mV/ms) in 191 unipolar electrograms recorded from ischemically damaged subepicardial muscle during programmed stimulation inducing reentrant tachycardias. Procainamide caused a greater reduction in /-dV/dt(max)/ than did lidocaine in the responses to basic stimulation, and it favored the occurrence of cycle length prolongation at tachycardia onset as the /-dV/dt(max)/ decreased progressively in successive beats. This resulted in conduction block and tachycardia termination in three of eight preparations. In contrast, lidocaine caused a greater depression in /-dV/dt(max)/ in response to closely coupled extrastimuli, but /-dV/dt(max)/ remained constant or even improved thereafter, and none of the tachycardias terminated spontaneously under lidocaine (n = 9). However, the reentrant circuits remained spatially unstable, and lidocaine favored the occurrence of cycle length dynamics displaying constant or decreasing trends. This study supports the notion that cycle length dynamics at tachycardia onset are determined by the properties of the reentrant substrate and their pharmacological modulation.     

A tale of two dogs: analyzing two models of canine ventricular electrophysiology

The extensive development of detailed mathematical models of cardiac myocyte electrophysiology in recent years has led to a proliferation of models, including many that model the same animal species and specific region of the heart and thus would be expected to have similar properties. In this paper we review and compare two recently developed mathematical models of the electrophysiology of canine ventricular myocytes. To clarify their similarities and differences, we also present studies using them in a range of preparations from single cells to two-dimensional tissue. The models are compared with each other and with new and previously published experimental results in terms of a number of their properties, including action potential morphologies; transmembrane currents during normal heart rates and during alternans; alternans onsets, magnitudes, and cessations; and reentry dynamics of spiral waves. Action potential applets and spiral wave movies for the two canine ventricular models are available online as supplemental material. We find a number of differences between the models, including their rate dependence, alternans dynamics, and reentry stability, and a number of differences compared with experiments. Differences between models of the same species and region of the heart are not unique to these canine models. Similar differences can be found in the behavior of two models of human ventricular myocytes and of human atrial myocytes. We provide several possible explanations for the differences observed in models of the same species and region of the heart and discuss the implications for the applicability of models in addressing questions of mechanism in cardiac electrophysiology.     

Temporal changes in N2O-losses from two arable soils

N₂O-loss rates from two soils were measured over a continuous observation period of 2 years. The two soils, differing in texture (sandy loam and silty loam), are frequently used for intensive crop production. Rates were estimated using a closed soil cover box technique. N₂O-losses obtained were scrutinised with physical, chemical and microbiological properties of the soils as well as with climatic data.Large temporal changes in N₂O-emission rates were found. The data were approximately log-normal distributed. In spring maximal values of 20 g N₂O-N ha^-1 d^-1 were observed. According to this observation, two situations associated with high flux rates could be distinguished; 1. N₂O- production by soil at spring thaw and 2. N₂O-production within one week after N-fertilizer application. For both soils equal N₂O-losses were found, which are adequate to 1 kg N₂O-N ha^-1 per year. From this data was calculated that N₂O-losses ranged from 0.8–1.5% of the applied fertilizer N.     

Increased 12-HETE production in bovine lymphocytes during selenium deficiency

When peripheral blood lymphocytes were incubated with arachidonic acid in the presence of Ca++ ionophore (A23187), the cells from the selenium-deficient dairy cows produced significantly greater quantities of 12-hydroxyeicosatetraenoic acid (12-HETE) than the cells from the selenium-supplemented animals. The major product of reaction was verified as 12-HETE by cochromatography with a 12-HETE standard on an HPLC and structural analysis by GC-MS. Additionally, concanavalin A-stimulated lymphocyte proliferation was significantly decreased in cells from the Se-deficient cows. Furthermore, 12-HETE generated by the A23187-stimulated lymphocytes inhibited lymphocyte proliferation when added to Se-supplemented cell cultures. These observations suggest that self-regulation of lymphocyte proliferation might be mediated by 12-HETE production, especially during an altered nutritional state such as Se deficiency.     

Temporal changes in allele frequencies in two reciprocally selected maize populations

The effects of breeding on allele frequency changes at 82 restriction fragment length polymorphism (RFLP) loci were examined in two maize (Zea mays L.) populations undergoing reciprocal recurrent selection, Iowa Stiff Stalk Synthetic and Iowa Corn Borer Synthetic #1. After 12 cycles of selection, approximately 30% of the alleles were extinct and 10% near fixation in each population. A test of selective neutrality identified several loci in each population whose allele frequency changes cannot be explained by genetic drift; interpopulation mean expected heterozygosity increased for that subset of 28 loci but not for the remaining 54 loci. Mean expected heterozygosity within the two subpopulations decreased 39%, while the between-population component of genetic variation increased from 0.5% to 33.4% of the total. Effective population size is a key parameter for discerning allele frequency changes due to genetic drift versus those resulting from selection and genetic hitchhiking. Empirical estimates of effective population size for each population were within the range predicted by the breeding method. Received: 10 June 1998 / Accepted: 29 April 1999     

Temporal properties of high frequency intra-QRS signals in myocardial infarction and healthy hearts.

The prevalence of late potentials after myocardial infarction depends on the site of the infarction. This may be caused by the different activation onsets of the anterior and inferior myocardial segments. Therefore, in anterior infarcts the high frequency signals may be concealed within the QRS whereas in the inferior infarcts they last beyond the end of the QRS. We compared the timing and the spatial patterns of high frequency intra-QRS signals (IQSs) in the different infarction sites. We investigated 14 patients with anterior infarcts, 17 patients with inferior infarcts, and 10 healthy subjects. 31-lead magnetocardiograms were recorded in left precordial position and averaged. The QRS signals were smoothed with a Savitzky-Golay filter. The smoothed QRS signals were subtracted from the measured ones. The difference of the signals (frequency band of about 60-200 Hz) representing the high frequency components was quantified. The percentage of the high frequency signals was calculated for the entire QRS, for the first and for the second half, respectively. We found that in patients with anterior infarcts the high frequency components predominantly appeared in the first half of the QRS whereas in inferior infarcts these components predominantly appeared in the second half of the QRS. The different infarction sites were associated with different spatial patterns of the high frequency signals on the body surface. In healthy subjects there was not such a preferential association of time intervals and high frequency signals. Late potentials are the special case of high frequency signals appearing in the terminal QRS. It is the general property of the myocardium to generate high frequency signals associated with the depolarization of infarcted tissue. The timing of such signals and the spatial distribution patterns on the body surface may help to identify the location of the sources.     

Temporal changes in the prevalence of parasites in two Oregon estuary-dwelling fishes

The parasite faunas of juvenile English sole (Parophrys vetulus) in 1971-1972 and staghorn sculpin (Leptocottus armatus) in 1971 from Yaquina Bay, Oregon, were compared with faunas found in the same estuary in 1997-2000 (English sole) and 1999-2000 (staghorn sculpin). The 7 most commonly occurring parasites in 1971 were compared with the same species observed during the same month and sampling sites in 1997-2000. Multivariate community analysis of juvenile English sole parasites supported the suggestion that the 1971 parasite data were representative of the early-1970s time period. Four of the parasite species infecting English sole and 6 of those infecting staghorn sculpins had significantly lower prevalences in 1997-2000. Parasite species with significantly lower prevalences also had reduced intensity levels. One parasite (Glugea stephani) of English sole increased in prevalence in the 1997-2000 samples in association with the warm estuarine temperatures during the 1997 El Ni?o year. Although the causes for the changes in occurrence of other parasites were not determined, ecological changes in Yaquina Bay that may have influenced parasite ecology include apparent changes in the estuary ichthyofauna that occurred between the sampling periods. Such changes could be associated with increases in the number of California sea lions (Zalophus californianus) subsequent to establishment of the Marine Mammal Protection Act in 1972.     

12-Hydroxyeicosatetraenoic acid (12-HETE) induces heat shock proteins in human leukocytes

12-Hydroxyeicosatetraenoic acid (12-HETE) induces the expression of individual heat shock proteins in human leukocytes (lymphocytes, monocytes, basophil granulocytes; LMBs). Metabolic radiolabeling of LMBs revealed that exogenous 12-HETE (20 ng) led to the expression of a 65- and 83kDa protein. Immunoreactivity towards the 65kDa protein was commonly detected. In contrast, after heat shock treatment and predominantly after incubation with 12-HETE significant immunoreactivity (anti-hsp72) was detected in the lower molecular weight range whereas immunoreactivity (anti-hsp90, AC88) was only observed after heat shock treatment of LMBs.     

Metabolism of 12(R)-hydroxy-5,8,10,14-eicosatetraenoic acid (12(R)-HETE) in corneal tissues: formation of novel metabolites

12(R)-Hydroxy-5,8,10,14-eicosatetraenoic acid [12(R)-HETE], a cytochrome P450 arachidonate metabolite, is metabolized by corneal tissues via three distinct metabolic pathways: beta-oxidation, omega-hydroxylation, and keto-reduction. The major metabolite released from the intact rabbit corneal epithelium or cultured cells was identified by mass spectrometric analysis as 8-hydroxy-4,6,10-hexadecatrienoic acid, the tetranor metabolite derived following two steps of beta-oxidation from the carboxy terminus. The beta-oxidation pathway was expressed in both microsomes and mitochondria isolated from bovine corneal epithelium and was dependent on the addition of oxidizing equivalents. The major metabolite of 12(R)-HETE in subcellular fractions of bovine corneal epithelial cells was a dihydro compound, 12-hydroxy-5,8,14-eicosatrienoic acid (12-HETrE). This derivative is presumably formed by an oxidation of the hydroxyl group followed by two keto-reduction steps, since its formation was accompanied by the appearance of a keto metabolite identified as 12-oxo-5,8,14-eicosatrienoic acid. The omega-hydroxylation, in contrast to other cell types, was a minor route for 12(R)-HETE metabolism in these tissues. Since 12(R)-HETE has been implicated as a modulator of Na(+)-K(+)-ATPase activity and its related functions in ocular tissues, these findings raise the possibility that the newly described metabolites may be involved in regulating corneal functions. In addition, the presence of a keto reductase in the cornea may be of great importance following injury since 12(R)-HETrE resulting from 12(R)-HETE by this activity is a potent ocular proinflammatory compound.