Istaroxime treatment ameliorates calcium dysregulation in a zebrafish model of phospholamban R14del cardiomyopathy.

The heterozygous Phospholamban p.Arg14del mutation is found in patients with dilated or arrhythmogenic cardiomyopathy. This mutation triggers cardiac contractile dysfunction and arrhythmogenesis by affecting intracellular Ca2+ dynamics. Little is known about the physiological processes preceding induced cardiomyopathy, which is characterized by sub-epicardial accumulation of fibrofatty tissue, and a specific drug treatment is currently lacking. Here, we address these issues using a knock-in Phospholamban p.Arg14del zebrafish model. Hearts from adult zebrafish with this mutation display age-related remodeling with sub-epicardial inflammation and fibrosis. Echocardiography reveals contractile variations before overt structural changes occur, which correlates at the cellular level with action potential duration alternans. These functional alterations are preceded by diminished Ca2+ transient amplitudes in embryonic hearts as well as an increase in diastolic Ca2+ level, slower Ca2+ transient decay and longer Ca2+ transients in cells of adult hearts. We find that istaroxime treatment ameliorates the in vivo Ca2+ dysregulation, rescues the cellular action potential duration alternans, while it improves cardiac relaxation. Thus, we present insight into the pathophysiology of Phospholamban p.Arg14del cardiomyopathy.

Steroid profile and IRMS analysis of musk administration for doping control.

Musk, the dried secretion of the musk pod (sac) of adult male musk deer, has been used as traditional Chinese Medicine (TCM) in China and south-east Asian countries for thousands of years. Due to the anabolic steroid component in this TCM, musk preparations have been included in the list of medical products containing prohibited substances employed for doping by the State Food and Drug Administration of China. The application of musk pod formulation was claimed to be responsible for some adverse analytical findings (AAFs) in the 2011 FIFA Women's World Cup. Our preliminary study has suggested that musk ingestion did not lead to AAFs of doping control with the single dosage of 100 mg. However, the influences of musk administration in large and multi dosage are still unclear. The aim of this study is to further investigate the influences of musk administration for doping control. Wild and domestic deer musk samples were collected. The concentrations and δ13 C-values of steroids in musk were analyzed. In an excretion study, 200 and 100 mg of wild and domestic deer musk samples were administrated by 29 subjects, respectively. Fluctuations in steroid profile could be observed, and the ratio of 5α-androstane-3α,17ß-diol to 5ß-androstane-3α,17ß-diol was more sensitive than other parameters. In the IRMS test, the ∆Δδ13 C-value between endogenous reference compound and etiocholanolone was a sensitive parameter, and AAFs were obtained. It is the first time to confirm with excretion study that musk administration could lead to positive result of doping control. Copyright © 2017 John Wiley & Sons, Ltd.

Agents with inotropic properties for the management of acute heart failure syndromes. Traditional agents and beyond.

Treatment with inotropic agents is one of the most controversial topics in heart failure. Initial enthusiasm, based on strong pathophysiological rationale and apparent empirical efficacy, has been progressively limited by results of controlled trials and registries showing poorer outcomes of the patients on inotropic therapy. The use of these agents remains, however, potentially indicated in a significant proportion of patients with low cardiac output, peripheral hypoperfusion and end-organ dysfunction caused by heart failure. Limitations of inotropic therapy seem to be mainly related to their mechanisms of action entailing arrhythmogenesis, peripheral vasodilation, myocardial ischemia and damage, and possibly due to their use in patients without a clear indication, rather than to the general principle of inotropic therapy itself. This review will discuss the characteristics of the patients with a potential indication for inotropic therapy, the main data from registries and controlled trials, the mechanism of the untoward effects of these agents on outcomes and, lastly, perspectives with new agents with novel mechanisms of action.

Istaroxime, a first in class new chemical entity exhibiting SERCA-2 activation and Na-K-ATPase inhibition: a new promising treatment for acute heart failure syndromes?

Acute heart failure syndromes (AHFS) are associated with the rapid onset of heart failure (HF) signs and symptoms. Hospitalizations for AHFS continue to rise and are associated with significant mortality and morbidity. Several pharmacological agents are currently approved for the treatment of AHFS, but their use is associated with an increase in short-term mortality. There is a need for new agents that can be given in the acute setting with increased efficacy and safety. Istaroxime is a unique agent with both inotropic and lusitropic properties which is currently being studied for the treatment of AHFS. Istaroxime inhibits the sodium-potassium adenosine triphosphatase (ATPase) and stimulates the sarcoplasmic reticulum calcium ATPase isoform 2 (SERCA-2) thereby improving contractility and diastolic relaxation. Early data from human studies reveal that istaroxime decreases pulmonary capillary wedge pressure (PCWP) and possibly improves diastolic function without causing a significant change in heart rate (HR), blood pressure, ischemic or arrhythmic events. Most commonly reported side effects were related to gastrointestinal intolerance and were dose related. In conclusion, istaroxime is a novel agent being investigated for the treatment of AHFS whose mechanism of action and cellular targets make it a promising therapy. Further studies with longer infusion times in patients with hypotension are required to confirm its efficacy and safety.

Hemodynamic, echocardiographic, and neurohormonal effects of istaroxime, a novel intravenous inotropic and lusitropic agent: a randomized controlled trial in patients hospitalized with heart failure.

OBJECTIVES: We examined the hemodynamic, echocardiographic, and neurohormonal effects of intravenous istaroxime in patients hospitalized with heart failure (HF). BACKGROUND: Istaroxime is a novel intravenous agent with inotropic and lusitropic properties related to inhibition of Na/K adenosine triphosphatase (ATPase) and stimulation of sarcoplasmic reticulum calcium ATPase. METHODS: One hundred twenty patients admitted with HF and reduced systolic function were instrumented with a pulmonary artery catheter within 48 h of admission. Three sequential cohorts of 40 patients each were randomized 3:1 istaroxime:placebo to a continuous 6-h infusion. The first cohort received 0.5 microg/kg/min, the second 1.0 microg/kg/min, and the third 1.5 microg/kg/min istaroxime or placebo. RESULTS: All doses of istaroxime lowered pulmonary capillary wedge pressure (PCWP), the primary end point (mean +/- SD: -3.2 +/- 6.8 mm Hg, -3.3 +/- 5.5 mm Hg, and -4.7 +/- 5.9 mm Hg compared with 0.0 +/- 3.6 mm Hg with placebo; p < 0.05 for all doses). Istaroxime significantly decreased heart rate (HR) and increased systolic blood pressure (SBP). Cardiac index increased and left ventricular end-diastolic volume decreased significantly only with 1.5 microg/kg/min. On echocardiography, left ventricular end diastolic volume and deceleration time improved with 1.5 microg/kg/min. There were no changes in neurohormones, renal function, or troponin I. Adverse events were not life threatening and were dose related. CONCLUSIONS: In patients hospitalized with HF, istaroxime improved PCWP and possibly diastolic function. In contrast to available inotropes, istaroxime increased SBP and decreased HR. (A Phase II Trial to Assess Hemodynamic Effects of Istaroxime in Pts With Worsening HF and Reduced LV Systolic Function [HORIZON-HF]; NCT00616161).

Rationale and design of the hemodynamic, echocardiographic and neurohormonal effects of istaroxime, a novel intravenous inotropic and lusitropic agent: a randomized controlled trial in patients hospitalized with heart failure (HORIZON-HF) trial.

BACKGROUND: Current inotropes have inodilator properties and, although are frequently used in acute heart failure syndromes, do not improve outcomes, likely from reduction in systolic blood pressure and increasing in arrhythmias, causing worsened myocardial ischemia and end-organ damage. Istaroxime is a novel agent that, in animal models, has both inotropic (inhibition of the Na/K ATPase channel) and lusitropic (stimulation of sarcoplasmic reticulum calcium ATPase activity) effects. HORIZON-HF is designed to test the hypothesis that istaroxime is effective in improving central hemodynamics and left ventricular (LV) function, without lowering systolic blood pressure, increasing heart rate, and worsening renal function or myocardial necrosis. METHODS AND RESULTS: This was a phase 2, randomized, double-blind, placebo-controlled, multicenter dose escalation exploratory study comparing 3 different doses of istaroxime to placebo in patients with LV systolic dysfunction (LV ejection fraction

Studies on the radioprotective effect of etiocholanolone.

One-hundred forty male mice of an inbred strain (C3H/St) were divided into four groups of thirty-five each and were given total body irradiation of 750 cGy. After X-irradiation, one group was given etiocholanolone alone, the other was given bone marrow alone, a third was given both of these. One group was used as a control and was given no treatment after irradiation. The bone marrow used was obtained from the same inbred strain of mice. The most remarkable hematologic recovery was seen in the group which was given both etiocholanolone and bone marrow. The groups given only etiocholanolone or only bone marrow recovered better than the control group. Overall survival was best for the group receiving both modes of treatment and worst for the control group. Survival for the groups receiving a single mode of treatment lay in between the other two mentioned above.

The effect of sustained delivery of androstanedione on the functional activities of adult male rats.

It is well documented that androgens stimulate protein anabolism, muscular development, bone growth and increased metabolic rate. In addition, exogenous low levels of testosterone can inhibit the secretion of gonadotropins by the anterior pituitary. The objective of this study was to investigate the effect of sustained delivery of androstanedione (AD) on the testicular architecture by means of tricalcium phosphate lysine (TCPL) delivery system. In this experiment adult male Sprague Dawley rats (250-300 g BW) were randomly divided into three equal groups (n = 16). Rats in group I were implanted subcutaneously with TCPL implants loaded with AD. Rats in group II were implanted with sham TCPL capsules, and rats in group III served as intact unimplanted controls. Surgical aseptic techniques were performed according to standard laboratory procedures. At the end of 2, 4, 8 and 16 weeks post implantation, four animals from each group were sacrificed and the testes were collected, weighted, and embedded for histopathological evaluations. The results of this study revealed the following: (1) remarkable reduction of testicular mass at the end of the 4 week phase and continued for the duration of study in comparison to the sham and intact control; (2) cross sections obtained from group I animals have shown that the Leydig cells were decreased in size and number of organelle; (3) the epithelium of the seminiferous tubules decreased in height and cell number; (4) a significant decrease (p < 0.05) in the number of primary and secondary spermatocytes (63% and 78% receptively) at the end of the 8 week phase, and azoospermia was observed at the 12 week phases in group I rats; and (5) slight reduction in the number of was observed in tissues obtained AD treated animals. This experiment confirms our previous findings using testosterone hormone and demonstrates that low sustained levels of exogenous AD can also impair fertility in rats.

Immunochemical and histopathological evaluation associated with sustained delivery of 5-alpha androstanedione using adult rats as a model.

The specific objective of this study was to evaluate the effect of androstanedione (dihydrotestosterone (DHT)), delivered in a sustained manner by tricalcium phosphate lysine (TCPL) delivery devices, on the biochemical and cytological architecture of reproductive organs using adult rats as a model. A total of twenty four rats were distributed randomly into three equal groups. Rats in group II were implanted (S/C) with empty TCPL devices and served as sham controls. Group III rats were implanted with 1.51 (250 micrograms/day) g/cm3 CDD containing 300 mg DHT each. Group I animals served as unimplanted controls at time zero. Upon sacrifice (6 weeks), the prostatic tissue were collected, fixed, embedded, and sectioned (H&E) by using standard lab protocols. Data analysis of prostatic tissue obtained from rats implanted with such low density TCPL showed microscopic appearance varied somewhat from one area of the gland to another, but generally, there was considerable hyperplasia, as evidenced by accentuation of glandular folds and an increase in the number and size of the epithelial cells, with some areas of the gland showing more severe hyperplasia than others. Meanwhile, cross-sections of prostate obtained from rats implanted with empty CDD were characterized by an essentially normal histological appearance of the gland. These data exemplify the complex interactions which occur between prostate components and exogenous sustained delivery of DHT that are reflected in cell structure and function. Biochemical analysis of the serum revealed that there is remarkable reduction in HDL, and IL 1 (33%). In contrast the level of IL 6 increased (50%) in experimental animals compared to the sham operated animals. In conclusion, this experiment demonstrates that TCPL capsules are capable of delivering supraphysiological doses of DHT in a sustained manner for 6 weeks. The results also reveal that doses of DHT slightly above the physiological level can result in systemic toxicity. In addition, supraphysiological levels of DHT could cause severe BNH and regression to spermatogenesis after 6 weeks in rats, and an increase in the risk factors associated with CVD.

The induction of interleukin-1 in humans and its metabolic effects.

To investigate the metabolic effects of interleukin-1 and its role as a mediator of host responses to trauma and sepsis, we injected seven healthy male subjects with etiocholanolone, an inflammatory agent that stimulates systemic responses thought to be mediated by interleukin-1. The subjects were fed a constant diet during each 4-day study and received three daily intramuscular injections of etiocholanolone, 0.10 mg/kg. Etiocholanolone injection resulted in inflammation, fever, leukocytosis, increased serum C-reactive protein, hypoferremia, and increased plasma activity of interleukin-1/lymphocyte-activating factor. Plasma concentrations of the counterregulatory hormones were normal. Protein metabolism, as reflected in nitrogen balance, 15N turnover, and forearm flux of alanine and glutamine, was unaltered. Serum glucose and insulin levels and tissue responsiveness to insulin were normal. This dissociation of acute-phase and catabolic responses may reflect the magnitude of the stimulus; higher levels of interleukin-1 may initiate catabolic responses. Alternatively, other mediators such as the counterregulatory hormones may direct the catabolic responses that occur after injury and sepsis.

[Changes in the concentration of luteinizing hormone in the blood serum and of luliberin in the hypothalamus of castrated rats after the administration of testosterone and its 5 alpha-reduced metabolites. I]. / Izmenenie kontsentratsii liuteiniziruiushchego gormona v syvorotke krovi i liuliberina v gipotalamuse kastrirovannykh krys posle vvedeniia testosterona i ego 5 al'fa-vosstanovlennykh metabolitov. Chast'I.

Luteinizing hormone concentration in the blood serum and luliberin content in the hypothalamic arcuate body and medial eminence were determined in castrated mature rats 1, 3, 6, 12, 24 hours afte single injections of testosterone and its 5 alpha-reduced metabolites (dihydrotestosterone, 5 alpha-androstane-3 alpha, 17 beta-diol and 5 alpha-androstane-3 beta, 17 beta-diol) at a dose of 2.5 x 10(-7) kg/kg body weight by radioimmunoassay. Principal differences between testosterone and its 5 alpha-reduced metabolite action on the hypothalamohypophyseal system were detected from the stand point of their effectiveness and the effect realization rate.

Correction of human cyclic neutropenia with prednisolone.

A 70-year-old woman with cyclic neutropenia was treated with 16 mg of etiocholanolone and 25 mg of prednisolone intramuscularly every other day. During 14 weeks' treatment amplitude of cyclic fluctuations in neutrophil counts gradually decreased, but pretreatment cycles returned promptly after treatment was stopped. Prednisolone alone every other day (25 mg) reproduced this result, and by 23 weeks, neutrophil counts became stable at about 1500 per cubic millimeter. tcycling of monocytes, platelets and reticulocytes was also eliminated, as were symptoms that had accompanied neutropenic periods. In addition, bone-marrow neutrophil precursors and neutrophil marrow reserves were stabilized. The patient was subsequently maintained satisfactorily with oral prednisolone, 20 mg every other day. These studies demonstrate that the discontinuous myeloid maturation that occurs in cyclic neutropenia can be corrected with prednisolone every other day.

Aetiocholanolone and prednisolone therapy in patients with severe bone-marrow failure.

Ten patients with bone-marrow failure from myelofibrosis, aplastic anaemia, paroxysmal nocturnal haemoglobinuria, and "hairy cell" leukaemia, who had failed to respond to androgens and/or glucocorticosteroids, were treated with a combination of aetiocholanolone and prednisolone. All patients except one with aplastic anaemia responded with an increase in reticulocytes, haematocrit, red blood cell mass, and blood neutrophils and platelets. These data suggest that aetiocholanolone, and similar 5beta-hydroxy steroids, may be valuable in treating certain patients with chronic marrow failure.

On the mechanism of erythropoietin-induced differentiation. XIV. The apparent effect of etiocholanolone on initiation of erythropoiesis.

Etiocholanolone, when tested in normal mice or in mice that have been out of an hypoxic atmosphere for only a few days, stimulates erythropoiesis, but appears to have no effect on erythropoiesis when tested in plethoric mice that have very low residual red cell formation. The steroid, similarly, stimulates hemoglobin synthesis by marrow cells in culture if they are from normal mice, but has no effect on cells from transfused mice. Out data suggest that 5 beta-H androstane and pregnane derivatives do not have a primary action on the induction of erythropoiesis, but act on cells that are already differentiated.