Case-control study of candidate gene methylation and adenomatous polyp formation.

PURPOSE: Colorectal cancer (CRC) is one of the most common and preventable forms of cancer but remains the second leading cause of cancer-related death. Colorectal adenomas are precursor lesions that develop in 70-90 % of CRC cases. Identification of peripheral biomarkers for adenomas would help to enhance screening efforts. This exploratory study examined the methylation status of 20 candidate markers in peripheral blood leukocytes and their association with adenoma formation. METHODS: Patients recruited from a local endoscopy clinic provided informed consent and completed an interview to ascertain demographic, lifestyle, and adenoma risk factors. Cases were individuals with a histopathologically confirmed adenoma, and controls included patients with a normal colonoscopy or those with histopathological findings not requiring heightened surveillance (normal biopsy, hyperplastic polyp). Methylation-specific polymerase chain reaction was used to characterize candidate gene promoter methylation. Odds ratios (ORs) and 95 % confidence intervals (95% CIs) were calculated using unconditional multivariable logistic regression to test the hypothesis that candidate gene methylation differed between cases and controls, after adjustment for confounders. RESULTS: Complete data were available for 107 participants; 36 % had adenomas (men 40 %, women 31 %). Hypomethylation of the MINT1 locus (OR 5.3, 95% CI 1.0-28.2) and the PER1 (OR 2.9, 95% CI 1.1-7.7) and PER3 (OR 11.6, 95% CI 1.6-78.5) clock gene promoters was more common among adenoma cases. While specificity was moderate to high for the three markers (71-97 %), sensitivity was relatively low (18-45 %). CONCLUSION: Follow-up of these epigenetic markers is suggested to further evaluate their utility for adenoma screening or surveillance.

Characterization of sinoatrial node in four conduction system marker mice.

The specialized cardiac conduction system (CCS) consists of the sinoatrial node (SAN) and the atrioventricular (AV) conduction system (AVCS), which includes proximal (AV node, bundle of His and bundle branches) and distal (Purkinje fibers) components. In four CCS marker mice [two transgenic (cGATA6|lacZ, CCS|lacZ) and two targeted gene knock-in (minK|lacZ, Hop|lacZ)] the expression of the lacZ gene (beta-gal) has been reported to mark portions of the proximal and distal AVCS; the expression of this marker in the adult SAN is unknown. The primary objective of this study was to analyze the utility of these marker mice in the identification of the SAN. Intercaval and interventricular septal regions, containing all the components of the CCS, were freshly dissected from adult mice based on the anatomical landmarks and sectioned. Immunohistochemical characterization was performed with SAN markers (Cx45, HCN4), compared to the reporter expression (beta-gal) and markers of the working myocardium (Cx40 and Cx43). In all four of the CCS marker mice, we found that beta-gal expression is consistently observed in the proximal and distal AVCS. However, the presence of lacZ gene expression in the working myocardium outside the CCS and/or the absence of this reporter expression in the SAN prevent the effective use of these mice to identify the SAN, leading us to conclude that none of the four CCS marker mice we studied specifically mark the SAN.

Melatonin metabolite excretion among cellular telephone users.

PURPOSE: The relationship between cellular telephone use and excretion of the melatonin metabolite 6-hydroxymelatonin sulfate (6-OHMS) was evaluated in two populations of male electric utility workers (Study 1, n=149; Study 2, n=77). MATERIALS AND METHODS: Participants collected urine samples and recorded cellular telephone use over 3 consecutive workdays. Personal 60-Hz magnetic field (MF) and ambient light exposures were characterized on the same days using EMDEX II meters. A repeated measures analysis was used to assess the effects of cellular telephone use, alone and combined with MF exposures, after adjustment for age, participation month and light exposure. RESULTS: No change in 6-OHMS excretion was observed among those with daily cellular telephone use >25 min in Study 1 (5 worker-days). Study 2 workers with >25 min cellular telephone use per day (13 worker-days) had lower creatinine-adjusted mean nocturnal 6-OHMS concentrations (p=0.05) and overnight 6-OHMS excretion (p=0.03) compared with those without cellular telephone use. There was also a linear trend of decreasing mean nocturnal 6-OHMS/creatinine concentrations (p=0.02) and overnight 6-OHMS excretion (p=0.08) across categories of increasing cellular telephone use. A combined effect of cellular telephone use and occupational 60-Hz MF exposure in reducing 6-OHMS excretion was also observed in Study 2. CONCLUSIONS: Exposure-related reductions in 6-OHMS excretion were observed in Study 2, where daily cellular telephone use of >25 min was more prevalent. Prolonged use of cellular telephones may lead to reduced melatonin production, and elevated 60-Hz MF exposures may potentiate the effect.

A GATA-6 gene heart-region-specific enhancer provides a novel means to mark and probe a discrete component of the mouse cardiac conduction system.

The transcriptional programs that specify the distinct components of the cardiac conduction system are poorly understood, in part due to a paucity of definitive molecular markers. In the present study we show that a cGATA-6 gene enhancer can be used to selectively express transgenes in the atrioventricular (AV) conduction system as it becomes manifest in the developing multichambered mouse heart. Furthermore, our analysis of staged cGATA-6/lacZ embryos revealed that the activity of this heart-region-specific enhancer can be traced back essentially to the outset of the cardiogenic program. We provide evidence that this enhancer reads medial/lateral and anterior/posterior positional information before the heart tube forms and we show that the activity of this enhancer becomes restricted at the heart looping stage to AV myocardial cells that induce endocardial cushion formation. We infer that a deeply-rooted heart-region-specific transcriptional program serves to coordinate AV valve placement and AV conduction system formation. Lastly, we show that cGATA-6/Cre mice can be used to delete floxed genes in the respective subsets of specialized heart cells.

Subfamilies of CR1 non-LTR retrotransposons have different 5'UTR sequences but are otherwise conserved.

CR1 elements and CR1-related (CR1-like) elements are a novel family of non-LTR retrotransposons that are found in all vertebrates (reptilia, amphibia, fish, and mammals), whereas more distantly related elements are found in several invertebrate species. CR1 elements have several features that distinguish them from other non-LTR retrotransposons. Most notably, their 3' termini lack a polyadenylic acid (poly A) tail and instead contain 2-4 copies of a unique 8 bp repeat. CR1 elements are present at approximately 100,000 copies in the chicken genome. The vast majority of these elements are severely 5' truncated and mutated; however, six subfamilies (CR1-A through CR1-F) are resolved by sequence comparisons. One of these subfamilies (i.e. CR1-B) previously was analyzed in detail. In the present study, we identified several full-length elements from the CR1-F subfamily. Although regions within the open reading frames and 3' untranslated regions of CR1-F and CR1-B elements are well conserved, their respective 5' untranslated regions are unrelated. Thus, our results suggest that new CR1 subfamilies form when elements with intact open reading frames acquire new 5' UTRs, which could, in principle, function as promoters.

Circadian and infradian rhythms in mood.

The aim of this study was to assess any variation in positive, negative and total affect recorded longitudinally; to compare the results with those from prior transverse or hybrid population studies, based on the same or a different method of mood rating; and to test for any association of mood with cardiovascular, hormonal and geophysical variables monitored concomitantly. The study approach was as follows. A clinically healthy 34-year-old man filled out the positive and negative affective scale (PANAS) questionnaire five times a day for 86 days. Systolic (S) and diastolic (D) blood pressure (BP) and heart rate (HR) were also measured automatically at 30-minute intervals with an ambulatory monitor from May 19 to June 29, 2000, while different endpoints of heart rate variability (HRV) were also determined at 5-minute intervals from beat-to-beat electrocardiogram (ECG) monitoring for 42 days between May 3 and June 14, 2000, with only short interruptions while the subject took a shower and changed ECG tapes. Saliva samples were collected at the times of mood ratings for one month for later determination of melatonin and cortisol concentrations. Intervals of 24 hours of the record of each variable displaced in increments of 24 hours were analyzed by chronobiologic serial section at a trial period of 24 hours to assess the circadian characteristics as they changed from one day to another. Estimates of the midline-estimating statistic of rhythm (MESOR) and circadian amplitude and acrophase obtained on consecutive days were correlated among variables to assess any associations. The findings were as follows. Overall, a circadian rhythm was demonstrated for all variables. A positive association was noteworthy between the circadian amplitude of negative affect and the MESOR of both SBP (r= 0.363; P= 0.029) and DBP (r= 0.389; P= 0.019), suggesting that BP is raised in the presence of large swings in negative affect. Needing further validation was a weak association between the MESOR of negative affect and the circadian amplitude of SBP (r= - 0.272; P = 0.108), suggesting a lowering of the circadian SBP amplitude in the presence of a strong negative affect. Of further interest was the lack of a statistically significant relation between positive and negative affect, not only in terms of the MESOR but also in terms of the circadian amplitude.

Melatonin metabolite levels in workers exposed to 60-Hz magnetic fields: work in substations and with 3-phase conductors.

Melatonin suppression by 50/60-Hz magnetic fields represents a plausible biological mechanism for explaining increased health risks in workers. Personal exposure to magnetic fields and ambient light, and excretion of the melatonin metabolite 6-hydroxymelatonin sulfate (6-OHMS), were measured over 3 consecutive workdays in electric utility workers. There was a magnetic field-dependent reduction in adjusted mean nocturnal and post-work 6-OHMS levels among men working more than 2 hours per day in substation and 3-phase environments and no effect among those working 2 hours or less. No changes were observed among men working in 1-phase environments. The results suggest that circular or elliptical magnetic field polarization, or another factor linked to substations and 3-phase electricity, is associated with magnetic field induced melatonin suppression in humans.

Modular regulation of cGATA-5 gene expression in the developing heart and gut.

The evolutionarily conserved GATA-5 transcription factor is an early and persistent marker of heart and gut development in diverse vertebrate species. To search for control regions that might regulate the chicken GATA-5 (cGATA-5) gene, we assayed a set of cGATA-5/lacZ constructs in transgenic mice and found evidence for two functionally conserved control regions that regulate different facets of cGATA-5 gene expression. The more distal control region is activated in embryonic endoderm at the head-fold stage, whereas the other control region contains a regulatory module that is activated in a restricted region of endoderm following closure of the gut tube. Remarkably, the latter control region also contains a complex regulatory module that is activated in the cardiac crescent at the head-fold stage and subsequently functions in several mesodermal components of the developing heart, including the outer (epicardial) layer. We discuss these results in terms of possible contributions of epicardial-derived cells to the formation of heart valves, conduction tissue, and compact myocardium. These transgenes thus reveal, and provide a means to further analyze, transcriptional programs for several facets of heart morphogenesis and gut development.

An Nkx-dependent enhancer regulates cGATA-6 gene expression during early stages of heart development.

The evolutionarily conserved GATA-6 transcription factor is an early and persistent marker of heart development in diverse vertebrate species. We previously found evidence for a functionally conserved heart-specific enhancer upstream of the chicken GATA-6 (cGATA-6) gene and in the present study we used transgenic mouse assays to further characterize this regulatory module. We show that this enhancer is activated in committed precursor cells within the cardiac crescent and that it remains active in essentially all cardiogenic cells through the linear heart stage. Although this enhancer can account for cGATA-6 gene expression early in the cardiogenic program, it is not able to maintain expression throughout the heart later in development. In particular, the enhancer is sequentially downregulated along the posterior to anterior axis, with activity becoming confined to outflow tract myocardium. Enhancers with similar properties have been shown to regulate the early heart-restricted expression of the mouse Nkx2.5 transcription factor gene. Whereas these Nkx2.5 enhancers are GATA-dependent, we show that the cGATA-6 enhancer is Nkx-dependent. We speculate that these enhancers are silenced to allow GATA-6 and Nkx2.5 gene expression to be governed by region-specific enhancers in the multichambered heart.

Geomagnetic disturbances are associated with reduced nocturnal excretion of a melatonin metabolite in humans.

The effects of geomagnetic disturbances on urinary excretion of the melatonin metabolite, 6-hydroxymelatonin sulfate (6-OHMS), were studied in conjunction with 60 Hz magnetic field (MF) and ambient light exposure in 132 electric utility workers. Geomagnetic activity was assessed using a local (equivalent amplitude or A(K), Boulder, CO) and global (average antipodal or aa) index. Personal exposures to 60 Hz MFs and light were obtained using data-logging meters. The relationship between geomagnetic activity and 6-OHMS was assessed with adjustment for age, light exposure, and month of participation. Mean overnight 6-OHMS excretion was lower on days when the 36-h A(K) or aa values exceeded 30 nT. A greater reduction in 6-OHMS excretion was observed when increased geomagnetic activity was combined with elevated 60 Hz MF or reduced ambient light exposures.

Reduced excretion of a melatonin metabolite in workers exposed to 60 Hz magnetic fields.

The effects of occupational 60 Hz magnetic field and ambient light exposures on the pineal hormone, melatonin, were studied in 142 male electric utility workers in Colorado, 1995-1996. Melatonin was assessed by radioimmunoassay of its metabolite, 6-hydroxymelatonin sulfate (6-OHMS), in post-work shift urine samples. Personal magnetic field and light exposures were measured over 3 consecutive days using EMDEX C meters adapted with light sensors. Two independent components of magnetic field exposure, intensity (geometric time weighted average) and temporal stability (standardized rate of change metric or RCMS), were analyzed for their effects on creatinine-adjusted 6-OHMS concentrations (6-OHMS/cr) after adjustment for age, month, and light exposure. Geometric mean magnetic field exposures were not associated with 6-OHMS/cr excretion. Men in the highest quartile of temporally stable magnetic field exposure had lower 6-OHMS/cr concentrations on the second and third days compared with those in the lowest quartile. Light exposure modified the magnetic field effect. A progressive decrease in mean 6-OHMS/cr concentrations in response to temporally stable magnetic fields was observed in subjects with low workplace light exposures (predominantly office workers), whereas those with high ambient light exposure showed negligible magnetic field effects. Melatonin suppression may be useful for understanding human biologic responses to magnetic field exposures.

Nocturnal excretion of a urinary melatonin metabolite among electric utility workers.

OBJECTIVES: The effects of 60-Hz magnetic field and ambient light exposures on the pineal hormone melatonin were studied among electric utility workers. METHODS: Personal exposure was measured at 15-second intervals over 3 consecutive 24-hour periods. Exposure metrics based on magnetic field intensity, intermittence, or temporal stability were calculated for periods of work, home, and sleep. A rate-of-change metric (RCM) was used to estimate intermittence, and the standardized RCM (RCMS = RCM/standard deviation) was used to evaluate temporal stability. The effects of magnetic field exposure on total overnight 6-hydroxymelatonin sulfate (6-OHMS) excretion and creatinine-adjusted nocturnal 6-OHMS (6-OHMS/cr) concentration were analyzed with adjustment for age, month, and light exposure. RESULTS: Magnetic field intensity, intermittence, or cumulative exposure had little influence on nocturnal 6-OHMS excretion. Residential RCMS magnetic field exposures were associated with lower nocturnal 6-OHMS/cr concentrations. In multivariate statistical analyses, the interaction term for geometric mean and RCMS magnetic field exposures at home was associated with lower nocturnal 6-OHMS/cr and overnight 6-OHMS levels. Modest reductions in the mean 6-OHMS levels occurred after RCMS exposures during work. The greatest reductions occurred when RCMS exposures both at work and at home were combined; therefore the effects of temporally stable magnetic fields may be integrated over a large portion of the day. CONCLUSIONS: Results from this study provide evidence that temporally stable magnetic field exposures are associated with reduced nocturnal 6-OHMS excretion in humans.

Common role for each of the cGATA-4/5/6 genes in the regulation of cardiac morphogenesis.

The GATA-4/5/6 genes encode transcription factors implicated previously in the regulation of cardiac-specific differentiation programs. However, recent analyses of mouse GATA-4 null mutations found evidence for function in endoderm development (in vitro) and embryonic morphogenesis (in vivo). Whether each of the three cardiac-associated GATA factors function within distinct or common developmental programs was previously untested; past studies defined specific and distinct roles for each of the GATA-1/2/3 genes in embryonic hematopoiesis. In this study, we compare the transcript patterns of cGATA-4/5/6 during chick embryogenesis. Each of the three GATA factors is expressed in a similar pattern within gastrulating cells of the primitive streak, prior to determination of the cardiomyocyte progenitors, and later within the lateral plate mesoderm and associated endoderm layer. The patterns overlap but extend beyond the presumptive cardiomyocyte population expressing cNkx-2.5. Later in development, cGATA-4/5/6 are all transcribed throughout the differentiating heart, in similar but not identical patterns, within the endocardium, myocardium, and great vessels. In order to test the function of GATA factors during chick cardiogenesis, embryos were cultured in vitro in the presence of antisense oligomers designed to deplete specifically transcripts encoding cGATA-4/5/6, beginning around stage 7. When oligomers are used to target transcripts for all three genes, a high percentage of the embryos develop abnormal hearts related to the failure to form a normal primitive heart tube. In the most severe phenotype, cardiac bifida results in two bilateral beating hearts. In some embryos, the paired heart primordia undergo partial fusion but fail to form a single looping heart tube. In all cases, cellular differentiation is not obviously affected, as the abnormal hearts form beating tissue. Depletion of transcripts encoding any single GATA factor, or any combination of two GATA factors, does not affect development. The partial depletion of all three genes in chick results in a remarkably similar phenotype compared to the null GATA-4 mutation in mouse. Therefore, in the chick, each of the GATA-4/5/6 genes functions in a common pathway, at the time of cardiac crescent formation, for regulating early embryonic cardiac morphogenesis, apparently associated with embryonic folding or the migration of primordia to form a primitive tube.

The chicken GATA-6 locus contains multiple control regions that confer distinct patterns of heart region-specific expression in transgenic mouse embryos.

The GATA-6 transcription factor is expressed in cardiogenic cells and during subsequent stages of heart development in diverse vertebrate species. To gain insights into the molecular events that govern this heart-restricted expression, we isolated the chicken GATA-6 gene and used several approaches to screen for associated control regions. Our analysis of two chicken GATA-6/lacZ constructs in transgenic mouse embryos was particularly revealing. One GATA-6/lacZ construct, which has 1.5 kilobase pairs of upstream sequences along with the promoter and first intron, was expressed exclusively in the atrioventricular canal region of the heart. This expression pattern is novel and appears to mark specialized myocardial cells that induce underlying endocardial cells to initiate valve formation. The other GATA-6/lacZ construct, which has an additional 7.7 kilobase pairs of upstream sequences, was expressed in the ventricle and outflow tract in addition to the atrioventricular canal. The failure of these GATA-6 control regions to function as enhancers in transfected cardiac myocyte cultures underscores the importance of using transgenic approaches to elucidate transcriptional controls that function in the developing heart. Although the endogenous GATA-6 gene is expressed throughout the heart, our results indicate that this is effected in a heart region-specific manner.

Chicken repeat 1 (CR1) elements, which define an ancient family of vertebrate non-LTR retrotransposons, contain two closely spaced open reading frames.

Chicken repeat 1 (CR1) elements comprise a family of non-long terminal repeat (LTR) retrotransposons that have several noteworthy features. For example, whereas most other non-LTR elements have poly(A) tracts or other simple A-rich repeats at their 3' ends, the 3' ends of CR1 elements conform to the consensus [(CATTCTRT)(GATTCTRT)1-3]. CR1 elements also display an unusual bias for severe 5' truncations: only approx. 30 (out of a total of approx. 30 000) CR1 elements in the chicken genome include significant portions of the pol-like open reading frame (ORF) that we previously identified and partially sequenced [Burch et al. (1993) Proc. Natl. Acad. Sci. USA 90, 8199-8203]. In the present study we derived a consensus sequence for this entire ORF (ORF2) as well as an upstream ORF (ORF1) and part of a 5' untranslated region (UTR). The conceptual translation product of ORF2 is predicted to contain an endonuclease domain in addition to a reverse transcriptase domain. These results suggest that CR1 elements retrotranspose using a "nick and prime" mechanism similar (but not identical) to other families of non-LTR elements.

Major differences between WHV and HBV in the regulation of transcription.

Studies were carried out to further characterize enhancer and promoter elements on the woodchuck hepatitis virus (WHV) genome. We were able to confirm the existence of WHV promoters analogous to the major promoters of the related human hepatitis B virus (HBV) and of an enhancer analogous to the recently described WHV E2 element (Ueda, K., Wei, Y., and Ganem, D., Virology 217, 413, 1996). However, we were unable to identity an enhancer analogous to the E1 element of (HBV), despite the fact that these two viruses share a high degree of sequence homology and genetic organization. Some factor binding sites in the E1 region appeared to be conserved between the two viruses and may be required for the activity of the overlapping X gene promoter of WHV. Others did not appear to be essential for WHV X gene promoter activity, and their functional activity, if any, was not revealed. Our failure to detect a functional enhancer element in the region of WHV homologous to the HBV E1 enhancer may indicate that (i) fundamental differences exist in transcriptional regulation of the small circular genomes of WHV and HBV; (ii) WHV contains an E1 element which is functional in the context of the intact viral genome, but which is unable to function in the context of the various expression constructs used in our experiments; or (iii) correct regulation of WHV transcription via an E1 element is dependent upon transcription factors which are not expressed in the liver-specific cell lines used in our experiments.

Transcripts for functionally distinct isoforms of chicken GATA-5 are differentially expressed from alternative first exons.

Our analysis of cDNA and genomic clones unexpectedly revealed that the chicken gata-5 gene is differentially expressed from alternative first exons. Moreover, we show that the respective transcripts are differentially processed to yield mRNAs for two distinct isoforms of GATA-5. The major isoform, which we described previously, has two CXNCX17CNXC zinc fingers typical of a vertebrate GATA factor. The minor isoform, on the other hand, has only one such zinc finger. We show that this novel isoform localizes within the nuclei of transfected cells and can bind to a consensus GATA site. This truncated isoform of GATA-5 is compromised in its ability to transactivate a simple target gene, however, and thus is functionally distinct from the major isoform of GATA-5.

A role for bone morphogenetic proteins in the induction of cardiac myogenesis.

Little is known about the molecular mechanisms that govern heart specification in vertebrates. Here we demonstrate that bone morphogenetic protein (BMP) signaling plays a central role in the induction of cardiac myogenesis in the chick embryo. At the time when chick precardiac cells become committed to the cardiac muscle lineage, they are in contact with tissues expressing BMP-2, BMP-4, and BMP-7. Application of BMP-2-soaked beads in vivo elicits ectopic expression of the cardiac transcription factors CNkx-2.5 and GATA-4. Furthermore, administration of soluble BMP-2 or BMP-4 to explant cultures induces full cardiac differentiation in stage 5 to 7 anterior medial mesoderm, a tissue that is normally not cardiogenic. The competence to undergo cardiogenesis in response to BMPs is restricted to mesoderm located in the anterior regions of gastrula- to neurula-stage embryos. The secreted protein noggin, which binds to BMPs and antagonizes BMP activity, completely inhibits differentiation of the precardiac mesoderm, indicating that BMP activity is required for myocardial differentiation in this tissue. Together, these data imply that a cardiogenic field exists in the anterior mesoderm and that localized expression of BMPs selects which cells within this field enter the cardiac myocyte lineage.

The chicken vitellogenin II gene is flanked by a GATA factor-dependent estrogen response unit.

The chicken vitellogenin II (VTGII) gene is flanked by an imperfect estrogen response element (ERE) at -350 and a perfect ERE at -620. In the present study we show that this imperfect ERE lies within an estrogen response unit (ERU) that requires a GATA factor and the estrogen receptor to function as an estrogen-dependent enhancer. We infer that GATA-6 contributes to the estrogen-dependent and liver-specific regulation of the endogenous VTGII gene since this is the predominant GATA factor expressed in adult liver. Our analysis of the VTGII ERU revealed four salient points. First, this ERU is comprised of an ERE and a bank of functionally redundant GATA-binding sites. Second, the GATA-6 transactivation domain is necessary (and sufficient, when tethered near the ERE) to render this ERU functional. Third, ERU enhancer activity is dependent on GATA 6, regardless of whether the resident ERE is imperfect or perfect. Fourth, in contrast to a report that the estrogen receptor antagonizes the activity of another GATA factor (GATA-1), we show that these two factors can function in a synergistic manner within the context of the VTGII ERU.

Random sequence phosphorothioate oligonucleotides evoke dramatic phenotypic alterations in cardiac myocyte cultures.

Cardiac myocytes displayed modest and uncoordinated contractile activity regardless of whether they are cultured in the presence or absence of unmodified random sequence oligonucleotides (oligos), as expected. Much to our surprise, however, when cardiac myocytes were cultured in the presence of random sequence phosphorothioate (PS) oligos, they reorganized into cablelike aggregates and displayed surging and coordinated contractile activity. Consistent with these observations, photobleaching experiments revealed that gap junction conductivity between affected cardiac myocytes was enhanced fourfold relative to control cultures. Furthermore, whereas atrial natriuretic factor (ANF) gene expression was induced in control cultures relative to intact hearts, this aberrant expression was selectively repressed in response to PS oligos. As PS oligos appear to mitigate deleterious effects that result from the proteolytic dispersal or culturing of cardiac myocytes or both, we suggest that these may useful cell culture reagents. It is interesting to contemplate whether cardiac myocytes might also be responsive to PS oligos within intact hearts, as this issue has potential clinical significance.