The ratio in the DNA content and protein mass of human cardiomyocytes

DNA and total protein content were measured in human cardiac myocytes using cytophotometry of Feulgen-naphthol yellow stained cells isolated from the left ventricle. The percentage of DNA classes was 2%, 32%, 53%, 12% and less than 0.5% for diploids, tetraploids (both 4c and 2c X 2), octaploids (8c, 4c X 2 and 2c X 4), hexadecaploids (16c, 8c X 2 and 4c X 4) and 32c cells, respectively. Binuclear cells comprised about 65% of the myocytes; the main class was 4c X 2. A discrepancy between total protein content and gene dose has been pointed out. DNA level ratio in a number of myocytes of different ploidy were 2:4:8:16:32; the same ratio for total protein content was 2:3.5:6:11.4:25.5, respectively.     

黄瓜四个品种的核型分析及其倍性变化的研究

本文对黄瓜的四个不同品种(“津一号”、“津二号”、“津四号”和“一串连”)进行了核型及倍性变化的研究,染色体数目为2n=14,其染色体组成为2n=2x=14=14m(2SAT)均为中部着丝点染色体,每一品种只有一对随体染色体,“津一号”和“津二号”还具有一个次级缢痕。黄瓜体细胞倍性的变化有3n、4n和6n,随着细胞倍性增高,其多倍性细胞频率依次下降,不同品种倍性的变化以“津四号”为最高,“一串连”品种为最低。     

Changes in binucleation and cellular dimensions of rat left atrial myocytes after induced left ventricular infarction

The left atrium of young rats has previously been demonstrated to respond with DNA synthesis and binucleation 11 days after left ventricular infarction. This investigation was designed to examine the hypertrophic response of the left atrial myocyte of the rat at 20 and 60 days after ventricular infarction. Male Sprague-Dawley rats were subjected to left coronary artery ligation (CAL) or sham operation. Following enzymatic separation, left atrial myocytes were examined at 20 and 60 days postoperation for number of nuclei and cellular dimensions (cell length, width and area, and nuclear area). Results demonstrated that the level of binucleation at 20 days (77.3%) and 60 days (71.3%) was nearly twice that observed in sham-operated animals, which were 33.1% binucleated at 20 days and 43.5% binucleated at 60 days. In both mononucleated and binucleated myocytes, the mean lengths, widths, and cell areas from CAL hearts were significantly greater than those of corresponding sham-operated animals. In all cases, these values were larger in binucleated myocytes than in mononucleated cells. The mean area of CAL cells was approximately twice that of sham-operated myocytes. With regard to mean lengths and widths, although both were greater in the CAL animals, there was a decrease in length and increase in width between 20 and 60 days in the CAL group. Mean nuclear areas were significantly greater in CAL myocytes than in those from the sham-operated group. These increases in nuclear number and cellular dimensions of the atrial myocyte are prominent features of the response to the stress imposed by left ventricular infarction.     

The ploidy variability of human cardiomyocytes

Unlike literature on ploidy of the human myocytes, DNA content and the number of nuclei have been revealed in the same cells. Besides, the cell ploidy were studied separately in the external, central and inner regions in the left ventricule. Considerable binucleation was revealed in normal and hypertrophic ventricles where 4c X2 was the modal cell class and there were 2c X 2, 8c X 2 and single 16c X 2 cells. Two alternative assumptions have been put forward concerning the genome differences: 1) myocytes enter in mitotic cycle under pathologic conditions or 2) the difference causes in genuine variability of the ploidy classes in normal hearts.     

Concomitant epicardial left atrial appendage ligation and left atrial ablation of atrial fibrillation: Safety,feasibility and outcome

IntroductionWe present initial results of patients undergoing a combined procedure of epicardial LAA ligation in addition to left atrial ablation for AF.Methods9 patients were included for additional use of LARIAT as an individual treatment approach for AF. First an epicardial LAA ligation was performed, in the same procedure left atrial ablations consisting of PVI and additional substrate based modifying ablations were performed. Follow–up at 3 months and 12 months was performed.ResultsThere was only 1 minor procedural complication (11%) involving epicardial bleeding and 2 late adverse events of pericardial tamponade and stroke. At the final follow-up (median 20 months) 7 patients were in stable sinus rhythm (78%) and 2 pts had reduced AF burden.ConclusionConcomitant epicardial LAA ligation and ablation is feasible in selected patients with a reasonable risk profile. More prospective data are required to validate the safety and efficacy.     

Functional significance of the left adrenal vein and the gonadal veins

Owing to the investigation of changeability variants of the left adrenal, anastomosis of the inferior diaphragmatic and ovarian (testicular) veins--tributaries of the human left renal vein, three variants in inflow of the left adrenal vein in comparison to the left ovarian (testicular) vein have been revealed. In the first variant, that occurs in 70% of cases, the left adrenal vein flows into the left renal vein by 15 cm more medially from the ostium of the ovarial (testicular) vein. The second variant occurs in 11%. The left adrenal and ovarian (testicular) ostia are situated at the same level. The distance from the hilus renalis to the place, where the ostia mentioned get into the renal vein, is about 39 mm. In the third variant (19%) the left adrenal vein gets into the left renal vein nearer to the hilus renalis than the ovarian (testicular) veins. An average distance from the hilus renalis to the ostium of the left adrenal vein is 33 mm. The second variant, when the left adrenal vein gets into the left renal vein, contributes to appearance of the primary varicocele at the left. When the operation for nephrectomy is made at the left, our data make it possible to recommend a purposeful ligation of the left renal vein more laterally to the ostium of the ovarian (testicular) vein. In 80% this ligation will not disturb the function of the left adrenal vein.     

Long-term effects of bone marrow mononuclear cell transplantation on left ventricular function and remodeling in rats

This study was performed to evaluate the long-term effect on left ventricular function and remodeling in a rat model of bone marrow cell transplantation (BMT) into acute infarcted myocardium. After myocardial infarction was induced in inbred Lewis rats by left anterior descending artery ligation, the ischemic area was directly injected with saline, peripheral blood mononuclear cells (PB-MNCs) or bone marrow mononuclear cells (BM-MNCs). Cardiac function and structure were evaluated by echocardiography before the operation, and on day 1 and 2 months post-infarct. The collagen content, the number of vessels and the vasculogenesis were examined by histology and immunohistochemistry. We found at 2 months post-infarct, BMT significantly improved cardiac systolic function and recovered diastolic function. Transplantation of BM-MNCs, but not PB-MNCs, reversed remodeling and reduced collagen density. Vessel counts showed greater angiogenesis occurred in the animals transplanted with BM-MNCs. Furthermore, a vascular endothelial cell-specific marker was detected in the transplanted bone marrow cells. Our data suggest that BM-MNC transplantation results in long-term improvement in left ventricular function-especially diastolic function- and remodeling, possibly related with the reduction of the amount of the collagen and enhancement of neovascularization.     

Heart innervation after ligation of the left anterior descending coronary artery (LAD)

Distribution and amount of neuropeptide Y- and synaptophysin-immunoreactive nervous structures within the heart were investigated in dogs 4 days after ligation of the left anterior descending coronary artery (LAD). In the right atrium and posterior left ventricular regions, which were taken as (non-infarcted) control areas, neuropeptide Y-immunoreactive paravascular nerves and a perivascular nerve plexus running within the adventitia of the coronary arteries and their branches down to the arterioles were observed. Morphometric measurements of the area density revealed 0.099 +/- 0.014% for synaptophysin- and 0.037 +/- 0.0072% for neuropeptide Y-immunoreactivity within the posterior wall of the left ventricular myocardium. Four days after ligation of the LAD only single synaptophysin- and neuropeptide Y-immunoreactive nerve fibers were very rarely detected in the infarcted region of the anterior wall of the left ventricle. Above the ligature larger than normal neuropeptide Y-immunoreactive axons within nerves along the LAD indicated a blockage of the axoplasmic transport of this peptide. When investigating this model of experimental myocardial infarction, mechanical traumatization of peri- and paravascular nerves of the LAD by the ligature has to be considered as a major pathogenetic factor, in addition to ischemia leading to denervation of infarcted as well as non-ischemic myocardium.     

Neurohormones in an ovine model of compensated postinfarction left ventricular dysfunction

Clinical heart failure, often the result of myocardial infarction, may be preceded by a period of compensated left ventricular impairment. There is substantial need for an experimental model that reflects this human condition. In sheep, coronary artery ligation produced consistent left ventricular anteroapical myocardial infarctions resulting in chronic (5 wk), stable hemodynamic changes compared with sham controls, including reductions in ejection fraction (51 +/- 2 vs. 30 +/- 5%, P < 0.001), cardiac output (6.3 +/- 0.2 vs. 5.1 +/- 0.2 l/min, P < 0.01), and arterial pressure (93 +/- 2 vs. 79 +/- 3 mmHg, P < 0.001), and increases in cardiac preload (left atrial pressure, 3.3 +/- 0.1 vs. 8.3 +/- 1.3 mmHg, P < 0.001). These changes were associated with acute and sustained increases in plasma concentrations of atrial natriuretic peptide (ANP; 5 wk, 11 +/- 2 vs. 27 +/- 5 pmol/l, P < 0.001), brain natriuretic peptide (BNP; 3 +/- 0.2 vs. 11 +/- 2 pmol/l, P < 0.001), and amino-terminal pro-brain natriuretic peptide (NT-BNP; 17 +/- 3 vs. 42 +/- 12 pmol/l, P < 0.001). Significant correlations were observed between plasma levels of the natriuretic peptides (ANP, day 7 to week 5 samples; BNP and NT-BNP, day 1 to week 5 samples) and changes in left ventricular volumes and ejection fraction. In contrast, renin activity, aldosterone, catecholamines, and endothelin were not chronically elevated postinfarction and were not related to indexes of ventricular function. Coronary artery ligation in sheep produces the pathological, hemodynamic, and neurohormonal characteristics of compensated left ventricular impairment secondary to myocardial infarction. Plasma concentrations of the cardiac natriuretic peptides are sensitive markers of left ventricular dysfunction. This is a reproducible model that reflects the clinical condition and should prove suitable for investigating the pathophysiology of, and experimental therapies in, early left ventricular dysfunction.     

Branches of the left pulmonary artery vascularizing the left upper pulmonary lobe

The studies were carried out on 100 left lungs taken from dead human bodies of both sexes whose age varied from 16 to 80 years. The pulmonary artery and the bronchus were injected with a 65% solution of duracryl and then digested in sulfuric acid. The specimens obtained were examined to determine the number and dimensions of the branches of the left pulmonary artery penetrating into the upper lobe of the left lung as well as the places at which they branch off from this artery. It was found that in most cases 4 branches ramified from the left pulmonary artery. Their length was 30 mm at the most, and their diameter, 12 mm. In about 50% of the cases the branches which penetrated into the lobe were the apicoanterior trunk, the lingular branch and 1 or 2 subsegmental branches, in about 25% of the cases almost all segmental branches penetrated into the lobe separately. In about 20% of the cases the apicoposterior trunk and independent segmental or subsegmental branches were present. Only in about 5% of the cases did the branches under consideration include the apicoposteroanterior trunk and the remaining segmental and subsegmental branches.     

Extravascular water content of heart and lungs after acute myocardial ischemia

We measured the extravascular water content of hearts and lungs of anesthetized dogs subjected to one of the following protocols: a)sham operation, b) circumflex artery ligation, c) increased left atrial pressure (Pla), or d) increased Pla and circumflex artery ligation. After 4 h, extravascular water of the heart and lungs increased significantly in the three experimental groups when compared with values from sham-operated dogs. After circumflex artery ligation, extravascular heart water increased 29% and lung water 8%, although Pla and calculated pulmonary microvascular pressure (Pmv) did not change. Extravascular heart water also rose 30% after increasing Pla from 23 to 37 cm H2O by inflating a left atrial baloon. In these dogs, extravascular lung water increased as a hyperbolic function of Pmv. Increasing Pla to 20 cm H2O in dogs with coronary artery ligation resulted in a 16% increase in heart water. Also at each Pmv, extravascular lung water was greater in dogs with coronary artery ligation than in dogs without. These data indicate that the increased extravascular lung water after coronary artery ligation cannot be explained solely by hemodynamic mechansims. We suggest that acute myocardial ischemia contributes to an increase in vascular permeability in the heart and lungs.     

Interferon alfa-2c in chronic myelogenous leukemia (CML): hematologic, cytogenetic and molecular-genetic response of patients with chronic phase CML previously resistant to therapy with interferon gamma

Alpha- and gamma-interferons have been shown to actively suppress hematopoiesis in patients in the chronic phase of chronic myelogenous leukemia in vitro and in vivo. Since both interferons act through different receptors on their hematopoietic target cells, they are expected to be capable of independently inhibiting abnormal blood cell development in patients with chronic myelogenous leukemia. We have utilized recombinant human interferon alfa-2c to treat 11 patients with Philadelphia chromosome positive chronic myelogenous leukemia in chronic phase, who were resistant to previous interferon gamma therapy. Ten of the patients were evaluable for hematologic, cytogenetic and molecular-genetic response following interferon alfa-2c therapy for 6 to 30 months. In 5 patients, IFN alfa-2c treatment failed due to lack of hematologic response. A complete hematologic or partial hematologic response was achieved in the remaining 5 patients. Three of these experienced cytogenetic improvement with reappearence of 100% diploid hematopoietic cells and disappearence of c-abl/bcr rearrangement in one patient. In two patients interferon alfa-2c did not prevent transformation of the disease into an accelerated state or blast crisis, respectively. We conclude that recombinant human interferon alfa-2c may also control hematopoiesis in interferon-gamma resistant chronic myelogenous leukemia patients, although the long-term response will need to be elucidated in further studies.     

Effect on breathing of abruptly loading and unloading the canine left heart

Cardiopulmonary bypass and pulmonary vein ligation were used to isolate left hearts of anesthetized open-chest dogs. After external gas exchange, blood was returned at constant flow (approximately 120 ml.min-1.kg-1) directly to the aorta or indirectly through the left heart ("left heart loading"). Loading caused breathing frequency (f) to increase approximately 5 breaths/min (approximately 20%), whereas systemic arterial pressure (Psa) fell approximately 15%. Because Psa was pulsatile during loading, we demonstrated separately the effect of pulsatile pressure and found it to lower mean Psa without changing f. Cooling cervical vagi to 7 degrees C eliminated the f response to loading and slightly decreased the Psa response. Loading was compared with graded distension of the fibrillating ventricle and beating atrium, which also increased f. As measured by an abdominal pneumograph, depth of breathing decreased significantly (approximately 4%) during left heart loading but did not change significantly on distension of the fibrillating heart. I conclude that left heart loading may induce tachypnea and a slightly reduced tidal volume by a vagal reflex most likely originating from the left heart.     

Effects of acute graded reductions in testicular blood flow on testicular morphology in the adult rat

The effect of moderate reductions in testicular blood flow has not been studied systematically. The aim of this study was, therefore, to examine the effects of different degrees of blood flow reduction on testicular morphology and to determine how much flow can be reduced before damage occurs. The subcapsular testicular artery was partially ligated in the left testes of adult rats. Testicular blood flow was measured before, immediately after, and 5 h after the ligation using laser Doppler flowmetry. After 5 h of partial ligation, the testes were removed, and their morphology was examined and related to the degree of blood flow reduction. The number of in situ end-labeled- or TUNEL-positive (i.e., dying) germ cells and the volume density of intravascular polymorphonuclear (PMN) leukocytes were measured. When flow was reduced to approximately 70% or less of its pretreatment value, a dose-related increase in the number of dying spermatogonia and early spermatocytes was seen. The PMN leukocytes accumulated in testicular blood vessels after partial ligation, and the maximum number was observed in testes where flow was reduced by approximately 50% of the pretreatment value. In conclusion, early stages of spermatogenesis are sensitive to a moderate, acute reduction in blood flow. Discrete reductions in flow may, therefore, have a large impact on sperm production.     

Kinetic features of a white rat hepatocyte population under hormonal disbalance before and after partial hepatectomy

Bilateral adrenalectomy, followed in 4 days by a partial hepatectomy, was performed using white rats weighing as much as 120-140 g. Under hormonal disbalance caused by bilateral adrenalectomy, the number of polyploid (4c, 4c x 2, and 8c) hepatocytes significantly increased, compared to that in non-operated control rats. Six hours after a partial hepatectomy, the share of highly ploid hepatocytes falls, being accompanied by a 9-fold increase in mitotic index. It is supposed that under hormonal disbalance condition, a partial hepatectomy may induce "early" mitoses in hepatocytes blocked in G2-phase of the cell cycle.     

Heart innervation after ligation of the left anterior descending coronary artery (LAD)

Summary Distribution and amount of neuropeptide Y- and synaptophysin-immunoreactive nervous structures within the heart were investigated in dogs 4 days after ligation of the left anterior descending coronary artery (LAD). In the right atrium and posterior left ventricular regions, which were taken as (non-infarcted) control areas, neuropeptide Y-immunoreactive paravascular nerves and a perivascular nerve plexus running within the adventitia of the coronary arteries and their branches down to the arterioles were observed. Morphometric measurements of the area density revealed 0.099±0.014% for synaptophysin- and 0.037±0.0072% for neuropeptide Y-immunoreactivity within the posterior wall of the left ventricular myocardium. Four days after ligation of the LAD only single synaptophysin-and neuropeptide Y-immunoreactive nerve fibers were very rarely detected in the infarcted region of the anterior wall of the left ventricle. Above the ligature larger than normal neuropeptide Y-immunoreactive axons within nerves along the LAD indicated a blockage of the axoplasmic transport of this peptide.When investigating this model of experimental myocardial infarction, mechanical traumatization of peri- and paravascular nerves of the LAD by the ligature has to be considered as a major pathogenetic factor, in addition to ischemia leading to denervation of infarcted as well as nonischemic myocardium.This study was supported by the SFB 320 within the Deutsche Forschungsgemeinschaft (DFG)Parts of the results were presented at the 59. Scientific Session of the American Heart Association (1986) and at the Congress of the Deutsche Gesellschaft für Herz- und Kreislaufforschung 1987.     

Phosphorylcholine antibodies restrict infarct size and left ventricular remodelling by attenuating the unreperfused post-ischaemic inflammatory response

Phosphorylcholine is a pro-inflammatory epitope exposed on apoptotic cells, and phosphorylcholine monoclonal immunoglobulin (Ig)G antibodies (PC-mAb) have anti-inflammatory properties. In this study, we hypothesize that PC-mAb treatment reduces adverse cardiac remodelling and infarct size (IS) following unreperfused transmural myocardial infarction (MI). Unreperfused MI was induced by permanent ligation of the left anterior descending (LAD) coronary artery in hypercholesterolaemic APOE*3-Leiden mice. Three weeks following MI, cardiac magnetic resonance (CMR) imaging showed a reduced LV end-diastolic volume (EDV) by 21% and IS by 31% upon PC-mAb treatment as compared to the vehicle control group. In addition, the LV fibrous content was decreased by 27% and LV wall thickness was better preserved by 47% as determined by histological analysis. Two days following MI, CCL2 concentrations, assessed by use of ELISA, were decreased by 81% and circulating monocytes by 64% as assessed by use of FACS analysis. Additionally, local leucocyte infiltration determined by immunohistological analysis showed a 62% decrease after three weeks. In conclusion, the local and systemic inflammatory responses are limited by PC-mAb treatment resulting in restricted adverse cardiac remodelling and IS following unreperfused MI. This indicates that PC-mAb holds promise as a therapeutic agent following MI limiting adverse cardiac remodelling.     

Seasonal changes in spermatogenesis in the blue fox (Alopex lagopus), quantified by DNA flow cytometry and measurement of soluble Mn2+ -dependent adenylate cyclase activity

The testes of the blue fox (Alopex lagopus) showed marked seasonal variations in size. Testicular weight and volume increased rapidly during January and February to reach maximal values by the beginning of the breeding season (approximately 15 March). During May and June the weights and volumes of the testes declined gradually to the quiescent state which lasted from July until October. Quantitation by DNA flow cytometry of the seasonal changes in the relative numbers of haploid (1C), diploid (2C) and tetraploid (4C) cell numbers in the testis showed that the increase in testis size from December to February was associated with a rapid expansion of the haploid cell compartment as spermatogenesis resumed. In addition, an increase in number of more mature cell types within the haploid cell population was observed over a 2-month period before the breeding season. The decline in testicular size from the middle of April until October was associated with a reduction in both the absolute and relative sizes of the haploid and tetraploid cell populations and a concomitant increase in the relative numbers of diploid cells. Measurements of the activity of the soluble Mn2+ -dependent adenylate cyclase revealed seasonal variations that closely paralleled those of the haploid cell population, indicating that, as in other species, the enzyme may be associated with maturing germ cells.     

Role of P2X purinergic receptors in the rescue of ischemic heart failure

Evidence is accumulating to support the presence of P2X purinergic receptors in the heart. However, the biological role of this receptor remains to be defined. The objectives here were to determine the role of cardiac P2X receptors in modulating the progression of post-myocardial infarction ischemic heart failure and to investigate the underlying mechanism. The P2X4 receptor (P2X4R) is an important subunit of native cardiac P2X receptors, and the cardiac-specific transgenic overexpression of P2X4R (Tg) was developed as a model. Left anterior descending artery ligation resulted in similar infarct size between Tg and wild-type (WT) mice (P > 0.1). However, Tg mice showed an enhanced cardiac contractile performance at 7 days, 1 mo, and 2 mo after infarction and an increased survival at 1 and 2 mo after infarction (P < 0.01). The enhanced intact heart function was manifested by a greater global left ventricular developed pressure and rate of contraction of left ventricular pressure in vitro and by a significantly increased fractional shortening and systolic thickening in the noninfarcted region in vivo (P < 0.05). The salutary effects on the ischemic heart failure phenotype were seen in both sexes and were not the result of any difference in infarct size in Tg versus WT hearts. An enhanced contractile function of the noninfarcted area in the Tg heart was likely an important rescuing mechanism. The cardiac P2X receptor is a novel target to treat post-myocardial infarction ischemic heart failure.