Post occlusive left atrial appendage thrombosis with extension into the left atrium.

A 67-year-old man underwent left atrial appendage (LAA) exclusion concomitant with mitral valve surgery and radiofrequency ablation maze procedure. On transoesophageal echocardiography anticipating ablation for left atrial tachycardia, an echodense thrombus was visualised in the LAA location with apparent intracavitary extension into the left atrium. Based on CT imaging findings, the echo represented thrombosis of a large left atrial appendage with probable extension into the left atrium.

Non-obstructive membranes of the left atrial appendage.

The left atrial appendage (LAA) is a blind-ending, complex structure distinct from the body of the left atrium and is sometimes regarded as a minor extension of the atrium. However, it should routinely be analysed as part of a transoesophageal echocardiographic examination. In this study we describe the presence of a non-obstructive membrane traversing the cavity of the LAA, found incidentally on transoesophageal echocardiography.

Impaired cardiac autophagy in patients developing postoperative atrial fibrillation.

OBJECTIVES: Postoperative atrial fibrillation (POAF) is a common complication after on-pump heart surgery. Several histologic abnormalities, such as interstitial fibrosis and vacuolization, have been described in atrial samples from patients developing POAF. This ultrastructural remodeling has been associated with the establishment of a proarrhythmic substrate. We studied whether atrial autophagy is activated in patients who develop POAF. METHODS: A total of 170 patients in sinus rhythm who had undergone elective coronary artery bypass grafting were included. Systemic inflammatory markers were measured at baseline and 72 hours after surgery. During the procedure, samples of the right atrial appendages were obtained for evaluation of remodeling by light and electron microscopy. Protein ubiquitination and autophagy-related LC3B processing were assessed by Western blot. RESULTS: Of these patients, 22% developed POAF. The level of high-sensitivity C-reactive protein, fibrosis, inflammation, myxoid degeneration, and ubiquitin-aggregates in the atria did not differ between patients with and without POAF. Electron microphotographs of those with POAF showed a significant accumulation of autophagic vesicles and lipofuscin deposits. Total protein ubiquitination was similar in the patients with and without POAF, but LC3B processing was markedly reduced in those with POAF, suggesting a selective impairment in autophagic flow. CONCLUSIONS: This study provides novel evidence that ultrastructural atrial remodeling characterized by an impaired cardiac autophagy is present in patients developing POAF after coronary artery bypass surgery.

Dual-enhanced cardiac CT for detection of left atrial appendage thrombus in patients with stroke: a prospective comparison study with transesophageal echocardiography.

BACKGROUND AND PURPOSE: A noninvasive method with high reliability and accuracy comparable to transesophageal echocardiography for identification of left atrial appendage thrombus would be of significant clinical value. The aim of this study was to assess the diagnostic performance of a dual-enhanced cardiac CT protocol for detection of left atrial appendage thrombi and for differentiation between thrombus and circulatory stasis in patients with stroke. METHODS: We studied 83 consecutive patients with stroke (56 men and 27 women; mean age, 62.6 years) who had high risk factors for thrombus formation and had undergone both dual-source CT and transesophageal echocardiography within a 3-day period. CT was performed with prospective electrocardiographic gating, and scanning began 180 seconds after the test bolus. RESULTS: Among the 83 patients, a total of 13 thrombi combined with spontaneous echo contrast and 14 spontaneous echo contrasts were detected by transesophageal echocardiography. All 13 thrombi combined with spontaneous echo contrast were correctly diagnosed on CT. Using transesophageal echocardiography as the reference standard, the overall sensitivity and specificity of CT for the detection of thrombi and circulatory stasis in the left atrial appendage were 96% (95% CI, 78% to 99%), and 100% (95% CI, 92% to 100%), respectively. On CT, the mean left atrial appendage/ascending aorta Hounsfield unit ratios were significantly different between thrombus and circulatory stasis (0.15 Hounsfield unit versus 0.27 Hounsfield unit, P=0.001). The mean effective radiation dose was 3.11 mSv. CONCLUSIONS: Dual-enhanced cardiac CT with prospective electrocardiographic gating is a noninvasive and sensitive modality for detecting left atrial appendage thrombus with an acceptable radiation dose.

Different structural remodelling in atrial fibrillation with different types of mitral valvular diseases.

AIMS: The aim of this study was to determine the relationship between atrial structural remodelling and atrial fibrillation (AF) for different types of mitral valvular diseases (MVDs). METHODS AND RESULTS: Left atrial appendages tissue samples were obtained from 24 patients with MVDs undergoing mitral valve replacement surgery. Masson's trichrome staining and immunohistochemical staining were performed to assess the extent of the fibrosis. Ultrastructural changes in left atrial appendages were examined by electron microscope. The degree of fibrosis showed significant increases in patients with AF compared with patients with sinus rhythm (SR) (P = 0.023). The collagen volume fraction (CVF) of fibrosis significantly increased in mitral stenosis and atrial fibrillation (MS-AF) compared with the mitral regurgitation and atrial fibrillation (MR-AF) group (P = 0.043). Collagen Type I levels were significantly increased in AF patients with mitral stenosis compared with AF patients with mitral regurgitation (P = 0.043). Different CVF of Matrix metalloproteinases-2 was present between the MS-SR group and the MS-AF group (P = 0.001). Electron microscopy revealed normally structured sarcomeres with a predominance of loosely packed cardiomyocytes in samples from patients with SR. Fibrotic bands, which tended to separate individual cardiomyocytes, were apparent in samples from patients with AF. CONCLUSION: Atrial structural remodelling is associated with AF patients with MVDs. Different heart rhythm statuses with different types of MVDs are associated with variable atrial structural remodelling. Different atrial structural remodelling is a mechanism that may contribute to the increased risk of AF with MVDs.

Ischemia triggers BNP expression in the human myocardium independent from mechanical stress.

BACKGROUND: It is unknown whether the increased B-type natriuretic peptide (BNP) values found in ischemic heart disease are triggered directly by ischemia or whether they are caused indirectly by ischemia through diastolic contractures or regional wall motion abnormalities. Therefore, we investigated the BNP expression in isolated human muscle strips under conditions of ischemia with and without mechanical stress. METHODS: Muscle strips (n=90) were isolated from human right atria (n=46). Contractures were induced by oxygen and glucose withdrawal. In 18 muscle strips contractures were prevented by means of butanedione monoxime (BDM). Sarcomere lengths were measured by electron microscopy (n=12). The gene expression and protein amount of BNP were determined and compared to control muscle strips contracting under physiological conditions. RESULTS: Hypoxia significantly decreased systolic force and induced diastolic contractures. This mechanical stress could be prevented in the group treated with BDM as evidenced by electron microscopy. Ischemia significantly increased BNP expression in both groups as evidenced by Northern blot analysis and immunohistochemistry. This increase was independent from mechanical stress. CONCLUSION: Our results indicate that ischemia is a potent mechanism for the expression of BNP. The increase in BNP expression under ischemic conditions is independent from concomitant mechanical alterations.

Atrial distribution of connexin 40 and 43 in patients with intermittent, persistent, and postoperative atrial fibrillation.

BACKGROUND: Sustained atrial fibrillation (AF) causes alterations in atrial electrical and structural properties. Conflicting data regarding the structural remodeling of the gap junction proteins connexin (Cx) 40 and 43 in human and animal studies exists. We investigated the amount and distribution of Cx40 and Cx43 in three subtypes of AF. METHODS: In 50 patients undergoing coronary artery bypass graft and/or mitral or aortic valve surgery, right atrial appendages were taken and examined with immunoconfocal microscopy. Retrospectively, four groups were built: (1) sinus rhythm pre- and postoperative (SR, n=20), (2) intermittent AF, but SR prior to surgery (intAF, n=6), (3) postoperative AF (popAF, n=12), and (4) persistent AF, at least 3 month prior to surgery (persAF, n=12). We analyzed the amount of Cx40 and Cx43 and the degree of fibrosis in three randomly selected areas of each sample. RESULTS: As compared with SR, the amount of Cx40 was significantly reduced by 53% in persAF. The distribution pattern of Cx40 was heterogeneous in patients with SR, intAF, and popAF, whereas patients with persAF showed similar densities of Cx40 in the three examined areas. We found no significant difference in the amount of Cx43 between the four groups. The distribution pattern of Cx43 was heterogeneous in all four groups. The Cx40/Cx43 ratio was significantly reduced in patients with popAF and persAF by 51% and 53%, respectively. No difference was seen in the degree of fibrosis between the four groups. CONCLUSIONS: In this study, sustained AF leads to a reduction in the amount of Cx40. Together with a specific Cx40/Cx43 ratio, this may contribute to localized conduction abnormalities, facilitating the self-perpetuation of re-entry pathways in AF. In the time course of structural atrial remodeling these changes seem to be earlier than a concomitantly developing fibrosis.

Myocyte changes and their left atrial distribution in patients with chronic atrial fibrillation related to mitral valve disease.

It has been found that the pulmonary veins and adjacent left atrial posterior wall (LAPW) are deeply involved in both the initiation and maintenance of atrial fibrillation (AF), and the identification of these high-risk sites has aroused great interest in investigating their histopathologic substrate. We used light and conventional electron microscopy to evaluate the differential myocyte and interstitial changes in LAPW and left atrial appendage (LAA) samples from 28 patients with chronic AF undergoing mitral valve surgery and from 12 autoptic controls. There were always more myocytes with loss of sarcomeres in the LAPW than in the LAA (19.9% +/- 7.7% versus 8.2% +/- 5.0%; P < .0001), and the LAPW showed more marked immunohistochemical evidence of dedifferentiation, characterized by the reexpression of smooth muscle actin. In pathological left atria, myocyte diameter in the LAPW and LAA was comparable (19.0 +/- 1.5 versus 18.5 +/- 2.0 microm; not significant) but larger than in the controls (11.9 +/- 0.8 and 12.1 +/- 1.3 microm, respectively; P < .0001). A terminal deoxynucleotidyltransferase assay did not reveal any myocyte apoptosis. The LAPW also showed more interstitial fibrosis than the LAA (7.49% +/- 3.34% versus 2.80% +/- 1.35%; P < .0001). Ultrastructural examination confirmed the presence of myocyte myocytolysis in the perinuclear area and showed changes in mitochondrial shape. In conclusion, the LAPW in patients with chronic AF related to mitral valve disease seems to be a particular anatomical site in which major myocyte and interstitial changes are concentrated, whereas the LAA is more protected. This remodeling may increase the heterogeneity of LAPW electrical conduction, thus confirming this location as an elective target for the ablation treatment of AF.

Peptidyl-glycine alpha-amidating monooxygenase targeting and shaping of atrial secretory vesicles: inhibition by mutated N-terminal ProANP and PBA.

ANP (atrial natriuretic peptide) is widely recognized as an important vasorelaxant, diuretic, and cardioprotective hormone. Little is known, however, about how ANP-secretory vesicles form within the atrial myocytes. Secretory vesicles were visualized by fluorescence microscope imaging in live rat atrial myocytes expressing proANP-enhanced green fluorescent protein (EGFP), or N-terminal-mutated fusion proteins thought to suppress the calcium-dependent aggregation of proANP. Results showed the following: (1) aggregates of proANP and coexpressed proANP-EGFP recruited peptidylglycine alpha-amidating monooxygenase (PAM)-1, an abundant atrial integral vesicle membrane protein; (2) coexpressed N-terminal-mutated (Glu23,24-->Gln23,24) and N-terminal-deleted proANP-EGFP inhibited recruitment of PAM-1 by up to 60%; (3) 4-phenyl-3-butenoic acid (PBA) (10 mumol/L), a pharmacological inhibitor of the lumenal peptidylglycine alpha-hydroxylating monooxygenase domain of PAM proteins, inhibited recruitment of endogenous PAM-1 and of coexpressed pro-EGFP-PAM-1; (4) PBA had no effect on exocytosis of the potassium inward rectifier KIR2.1; (5) PBA induced a deformation of the secretory vesicles but did not inhibit docking. These findings suggest that recruitment of PAM-1 to secretory vesicles depends on intact N-terminal proANP and on the lumenal domain of PAM-1. Conversely, PAM-1 participates in shaping the proANP-secretory vesicles. The full text of this article is available online at http://circres.ahajournals.org.

Atrial fibrillation is associated with increased spontaneous calcium release from the sarcoplasmic reticulum in human atrial myocytes.

BACKGROUND: Spontaneous Ca2+ release from the sarcoplasmic reticulum (SR) can generate afterdepolarizations, and these have the potential to initiate arrhythmias. Therefore, an association may exist between spontaneous SR Ca2+ release and initiation of atrial fibrillation (AF), but this has not yet been reported. METHODS AND RESULTS: Spontaneous Ca2+ release from the SR, manifested as Ca2+ sparks and Ca2+ waves, was recorded with confocal microscopy in atrial myocytes isolated from patients with and those without AF. In addition, the spontaneous inward current associated with Ca2+ waves was measured with the use of the perforated patch-clamp technique. The Ca2+ spark frequency was higher in 8 patients with AF than in 16 patients without (6.0+/-1.2 versus 2.8+/-0.8 sparks/mm per second, P<0.05). Similarly, the spontaneous Ca2+ wave frequency was greater in patients with AF (2.8+/-0.5 versus 1.1+/-0.3 waves/mm per second, P<0.01). The spontaneous inward current frequency was also higher in 10 patients with AF than in 13 patients without this arrhythmia (0.101+/-0.028 versus 0.031+/-0.007 per second, P<0.05, at a clamped potential of -80 mV). In contrast, both the Ca2+ released from the SR and the Na+-Ca2+ exchange rate induced by a rapid caffeine application were comparable in patients with and without AF. CONCLUSIONS: The observed increase in spontaneous Ca2+ release in patients with AF probably is due to an upregulation of the SR Ca2+ release channel activity, which may contribute to the development of AF.

Calpains and cytokines in fibrillating human atria.

Atrial fibrillation (AF) is accompanied by intracellular calcium overload. The purpose of this study was to assess the role of calcium-dependent calpains and cytokines during AF. Atrial tissue samples from 32 patients [16 with chronic AF and 16 in sinus rhythm (SR)] undergoing open heart surgery were studied. Atrial expression of calpain I and II, calpastatin, troponin T (TnT), troponin C (TnC), and cytokines [interleukin (IL)-1 beta, IL-2, IL-6, IL-8, IL-10, transforming growth factor (TGF)-beta 1, and tumor necrosis factor-alpha] were determined. Expression of calpain I was increased during AF (461 +/- 201% vs. 100 +/- 34%, P < 0.05). Amounts of calpain II and calpastatin were unchanged. Total calpain enzymatic activity was more than doubled during AF (35.2 +/- 17.7 vs. 12.4 +/- 9.2 units, P < 0.05). In contrast to TnC, TnT levels were reduced in fibrillating atria by 26% (P < 0.05), corresponding to the myofilament disintegration seen by electron microscopy. Small amounts of only IL-2 and TGF-beta 1 mRNA and protein were detected regardless of the underlying cardiac rhythm. In conclusion, atria of patients with permanent AF show evidence of calpain I activation that might contribute to structural remodeling and contractile dysfunction, whereas there is no evidence of activation of tissue cytokines.

Ultrastructural and morphometric features of nodal and impulse-conducting cardiac myocytes of the bat Pipistrellus pipistrellus.

Cells of the impulse-generating and conducting tissues of the insect-eating bat Pipistrellus pipistrellus were studied and evaluated using ultrastructural morphometry. Sinoatrial node cells are smaller than working atrial cells and measure about 6.5 microm in diameter. Their mitochondira and myofibril content constitute 23% and 19% of cytoplasmic volume, respectively. Corresponding values for working atrial cells are 23% and 52%. Atrioventricular node cells are 4.2 microm in diameter and contain abundant glycogen in the cytoplasm. The fractional volume of mitochondria in about 24% while that of myofibrils is 7%. Cells of the bundle of His are larger (6-8 microm diameter) and contain more cellular organelles than do nodal cells. Their mitochondria and myofibril contents are 25% and 25%, respectively. Cells in the proximal part of the right bundle branch are slender with diameters averaging 3.4 microm. Mitochondrial content is 23% while myofibrils occupy 20% of the cytoplasmic volume of these cells. Distally located bundle branch cells measure 7-10 microm in diameter with mitochondria and myofibril volumes of 30% and 33%. Subendocardial cells in the ventricular free wall are large reaching 28 microm in diameter (cf. 14-18 microm in working ventricular cells) and have mitochondira and myofibril volume fractions of 32% and 29%, respectively (35% & 40% for working ventricular cells).