Mechanisms of adaptive hypertrophic cardiac remodeling in a large animal model of premature ventricular contraction-induced cardiomyopathy.

Frequent premature ventricular contractions (PVCs) promoted eccentric cardiac hypertrophy and reduced ejection fraction (EF) in a large animal model of PVC-induced cardiomyopathy (PVC-CM), but the molecular mechanisms and markers of this hypertrophic remodeling remain unexplored. Healthy mongrel canines were implanted with pacemakers to deliver bigeminal PVCs (50% burden with 200-220 ms coupling interval). After 12 weeks, left ventricular (LV) free wall samples were studied from PVC-CM and Sham groups. In addition to reduced LV ejection fraction (LVEF), the PVC-CM group showed larger cardiac myocytes without evident ultrastructural alterations compared to the Sham group. Biochemical markers of pathological hypertrophy, such as store-operated Ca2+ entry, calcineurin/NFAT pathway, ß-myosin heavy chain, and skeletal type α-actin were unaltered in the PVC-CM group. In contrast, pro-hypertrophic and antiapoptotic pathways including ERK1/2 and AKT/mTOR were activated and/or overexpressed in the PVC-CM group, which appeared counterbalanced by an overexpression of protein phosphatase 1 and a borderline elevation of the anti-hypertrophic factor atrial natriuretic peptide. Moreover, the potent angiogenic and pro-hypertrophic factor VEGF-A and its receptor VEGFR2 were significantly elevated in the PVC-CM group. In conclusion, a molecular program is in place to keep this structural remodeling associated with frequent PVCs as an adaptive pathological hypertrophy.

Alterations of sarcoplasmic reticulum-mediated Ca2+ uptake in a model of premature ventricular contraction (PVC)-induced cardiomyopathy.

Premature ventricular contractions (PVCs) are the most frequent ventricular arrhythmias in the overall population. PVCs are known to acutely enhance contractility by the post-extrasystolic potentiation phenomenon, but over time persistent PVCs promote PVC-induced cardiomyopathy (PVC-CM), characterized by a reduction of the left ventricular (LV) ejection fraction. Ca2+ cycling in myocytes commands muscle contraction and in this process, SERCA2 leads the Ca2+ reuptake into the sarcoplasmic reticulum (SR) shaping cytosolic Ca2+ signal decay and muscle relaxation. Altered Ca2+ reuptake can contribute to the contractile dysfunction observed in PVC-CM. To better understand Ca2+ handling using our PVC-CM model (canines with 50% PVC burden for 12 weeks), SR-Ca2+ reuptake was investigated by measuring Ca2+ dynamics and analyzing protein expression. Kinetic analysis of Ca2+ reuptake in electrically paced myocytes showed a ~ 21 ms delay in PVC-CM compared to Sham in intact isolated myocytes, along with a ~ 13% reduction in SERCA2 activity assessed in permeabilized myocytes. Although these trends were not statistically significant between groups using hierarchical statistics, relaxation of myocytes following contraction was significantly slower in PVC-CM vs Sham myocytes. Western blot analyses indicate a 22% reduction in SERCA2 expression, a 23% increase in phospholamban (PLN) expression, and a 50% reduction in PLN phosphorylation in PVC-CM samples vs Sham. Computational analysis simulating a 20% decrease in SR-Ca2+ reuptake resulted in a ~ 22 ms delay in Ca2+ signal decay, consistent with the experimental result described above. In conclusion, SERCA2 and PLB alterations described above have a modest contribution to functional adaptations observed in PVC-CM.

Characterization of free fatty acid receptors expression in an obesity rat model with high sucrose diet.

INTRODUCTION/AIMS: In recent years, it has been shown that free fatty acids receptors (FFAR) of whose function in the cell surface plays a significant role in the regulation of cell function and nutrition as well are activated by various endogenous ligands, but mainly by fatty acids. Within FFAR of our interest are GPR 41, 43 and 120. The functions of these receptors are varied and dependent on the tissue where they are. The activation and signaling of these receptors, FFAR, are involved in many physiological processes, and currently the target of many drugs in metabolic disorders like obesity, diabetes and atherosclerosis. MATERIAL AND METHODS: Obesity was induced with hypercaloric diet (HD) in male Wistar rats for 20 weeks (n = 10). At the end, adipose tissue (abdominal and subcutaneous) was taken to perform assays for relative quantification mRNA expression by end-point RT-PCR and protein level expression by Western blot. RESULTS: These present data have shown for the first time that total mRNA isolation and protein expression from both adipose tissues (abdominal and subcutaneous) of rat in obesity condition yield significative statistical difference among the control versus obese groups, showing that the diet high in carbohydrates modifies the total presence of mRNA and protein level expression of the receptors GPR41, 43 and 120. CONCLUSIONS: Further comparative methods are in process to clarify whether or not the obesity changes the functional receptors in these two tissues for new pharmacological approaches.

Effects of topical capsaicin combined with moderate exercise on insulin resistance, body weight and oxidative stress in hypoestrogenic obese rats.

BACKGROUND: Hypoestrogenic (HE) women are one of the most vulnerable groups for the development of obesity and its complications. Capsaicin and exercise have demonstrated to reduce body weight and to improve insulin sensitivity in different animal models, but it is unknown whether their combination could be useful in HE obese females. METHODS: We investigated whether topical capsaicin, exercise or their combination had better therapeutic effects in an obesity-hypoestrogenism model. Ovariectomized Wistar rats were given a 30% sucrose solution (HE-Obese (HEOb)) or purified water (HE) during 28 weeks ad libitum; four experimental groups per each condition. After shaving the abdominal skin, cold cream vehicle was applied to the Sedentary groups (Sed) and capsaicin cream 0.075% (0.6 mg kg-1 per day) to the Capsaicin groups (Cap). Exercise (Ex) groups ran on a treadmill every day for 20 min at speeds from 9 to 18 m per min increased every 10 days; combination groups (Cap+Ex) were given topical capsaicin 90 min before exercise. The treatments were performed for 6 weeks, and caloric intake and body weight were monitored. At the end of the experimental protocol, glucose tolerance tests were performed, the animals were killed by decapitation; blood and organs were obtained to perform oxidative profile, histology, biochemical analyses and Western blot. RESULTS: In HEOb rats, the combined therapy reduced caloric intake, body weight and abdominal fat in a higher proportion than the individual treatments; it also decreased insulin resistance (IR), oxidative stress and pancreatic islet size. It was the only treatment that significantly increased p-AMPK levels in the soleus muscle. In HE rats, topical capsaicin was the only treatment that reduced glucose intolerance and improved the oxidative profile in a higher proportion than the combined therapy or Ex alone. CONCLUSIONS: Capsaicin per se or its combination with moderate exercise could be a useful therapy against complications linked to obesity-IR in HE females.

Proteome of Agave angustifolia Haw.: Uncovering metabolic alterations, over-accumulation of amino acids, and compensatory pathways in chloroplast-deficient albino plantlets.

Amino acids (AA) are essential molecules for plant physiology, acting as precursor molecules for proteins and other organic compounds. Chloroplasts play a vital role in AA metabolism, yet little is known about the impact on AA metabolism of albino plants' lack of chloroplasts. In this study, we conducted a quantitative proteome analysis on albino and variegated somaclonal variants of Agave angustifolia Haw. to investigate metabolic alterations in chloroplast-deficient plants, with a focus on AA metabolic pathways. We identified 82 enzymes involved in AA metabolism, with 32 showing differential accumulation between the somaclonal variants. AaCM, AaALS, AaBCAT, AaIPMS1, AaSHMT, AaAST, AaCGS, and AaMS enzymes were particularly relevant in chloroplast-deficient Agave plantlets. Both variegated and albino phenotypes exhibited excessive synthesis of AA typically associated with chloroplasts (aromatic AAs, BCAAs, Asp, Lys, Pro and Met). Consistent trends were observed for AaBCAT and AaCM at mRNA and protein levels in albino plantlets. These findings highlight the critical activation and reprogramming of AA metabolic pathways in plants lacking chloroplasts. This study contributes to unraveling the intricate relationship between AA metabolism and chloroplast absence, offering insights into survival mechanisms of albino plants.

Rate of onset of dopamine transporter inhibitors assessed with intracranial self-stimulation and in vivo dopamine photometry in rats.

Drug self-administration and intracranial self-stimulation (ICSS) are two preclinical behavioral procedures used to predict abuse potential of drugs, and abuse-related drug effects in both procedures are thought to depend on increased mesolimbic dopamine (DA) signaling. Drug self-administration and ICSS yield concordant metrics of abuse potential across a diverse range of drug mechanisms of action. The "rate of onset," defined as the velocity with which a drug produces its effect once administered, has also been implicated as a determinant of abuse-related drug effects in self-administration procedures, but this variable has not been systematically examined in ICSS. Accordingly, this study compared ICSS effects produced in rats by three DA transporter inhibitors that have different rates of onset (fastest to slowest: cocaine, WIN-35428, RTI-31) and that produced progressively weaker metrics of abuse potential in a drug self-administration procedure in rhesus monkeys. Additionally, in vivo photometry using the fluorescent DA sensor dLight1.1 targeted to the nucleus accumbens (NAc) was used to assess the time course of extracellular DA levels as a neurochemical correlate of behavioral effects. All three compounds produced ICSS facilitation and increased DA levels assessed by dLight. In both procedures, the rank order of onset rate was cocaine > WIN-35428 > RTI-31; however, in contrast to monkey drug self-administration results, maximum effects did not differ across compounds. These results provide additional evidence that drug-induced increases in DA drive ICSS facilitation in rats and illustrate the utility of both ICSS and photometry to evaluate the time course and magnitude of abuse-related drug effects in rats.

Strategies for the startup of methanogenic inverse fluidized-bed reactors using colonized particles.

One of the inconveniences in the startup of methanogenic inverse fluidized-bed reactors (IFBRs) is the long period required for biofilm formation and stabilization of the system. Previous researchers have preferred to start up in batch mode to shorten stabilization times. Much less work has been done with continuous-mode startup for the IFBR configuration of reactors. In this study, we prepared two IFBRs with similar characteristics to compare startup times for batch- and continuous-operation modes. The reactors were inoculated with a small quantity of colonized particles and run for a period of 3 months, to establish the optimal startup strategy using synthetic media as a substrate (glucose as a source of carbon). After the startup stage, the continuous- and batch-mode reactors removed more than 80% of the chemical oxygen demand (COD) in 51 and 60 days of operation, respectively; however, at the end of the experiments, the continuous-mode reactor had more biomass attached to the support media than the batch-mode reactor. Both reactors developed fully covered support media, but only the continuous-mode reactor had methane yields close to the theoretical value that is typical of stable reactors. Then, a combined startup strategy was proposed, with industrial wastewater as the substrate, using a sequence of batch cycles followed by continuous operation, which allows stable operation at an organic loading rate of 20 g COD/L x d in 15 days. Using a fraction of colonized support as an inoculum presents advantages, with respect to previously reported strategies.

Biogas production from the mechanically pretreated, liquid fraction of sorted organic municipal solid wastes.

The high liquid content in fruit and vegetable wastes makes it convenient to mechanically separate these wastes into mostly liquid and solid fractions by means of pretreatment. Then, the liquid fraction can be treated using a high-rate anaerobic biofilm reactor to produce biogas, simultaneously reducing the amount of solids that must be landfilled. In this work, the specific composition of municipal solid waste (MSW) in a public market was determined; then, the sorted organic fraction of municipal solid waste was treated mechanically to separate and characterize the mostly liquid and solid fractions. Then, the mesophilic anaerobic digestion for biogas production of the first fraction was evaluated. The anaerobic digestion resulted in a reduced hydraulic retention time of two days with high removal of chemical oxygen demand, that is, 88% on average, with the additional benefit of reducing the mass of the solids that had to be landfilled by about 80%.

Design, synthesis, and structure-activity relationships of a series of 3-[2-(1-benzylpiperidin-4-yl)ethylamino]pyridazine derivatives as acetylcholinesterase inhibitors.

Starting from the 3-[2-(1-benzylpiperidin-4-yl)ethylamino]-6-phenylpyridazine 1, we performed the design, the synthesis, and the structure-activity relationships of a series of pyridazine analogues acting as AChE inhibitors. Structural modifications were achieved on four different parts of compound 1 and led to the following observations: (i) introduction of a lipophilic environment in the C-5 position of the pyridazine ring is favorable for the AChE-inhibitory activity and the AChE/BuChE selectivity; (ii) substitution and various replacements of the C-6 phenyl group are possible and led to equivalent or slightly more active derivatives; (iii) isosteric replacements or modifications of the benzylpiperidine moiety are detrimental to the activity. Among all derivatives prepared, the indenopyridazine derivative 4g was found to be the more potent inhibitor with an IC(50) of 10 nM on electric eel AChE. Compared to compound 1, this represents a 12-fold increase in potency. Moreover, 3-[2-(1-benzylpiperidin-4-yl)ethylamino]-5-methyl-6-phenylpyridazine 4c, which showed an IC(50) of 21 nM, is 100-times more selective for human AChE (human BuChE/AChE ratio of 24) than the reference compound tacrine.

Aminopyridazines as acetylcholinesterase inhibitors.

Following the discovery of the weak, competitive and reversible acetylcholinesterase (AChE)-inhibiting activity of minaprine (3c) (IC50 = 85 microM on homogenized rat striatum AChE), a series of 3-amino-6-phenylpyridazines was synthesized and tested for inhibition of AChE. A classical structure-activity relationship exploration suggested that, in comparison to minaprine, the critical elements for high AChE inhibition are as follows: (i) presence of a central pyridazine ring, (ii) necessity of a lipophilic cationic head, (iii) change from a 2- to a 4-5-carbon units distance between the pyridazine ring and the cationic head. Among all the derivatives investigated, 3-[2-(1-benzylpiperidin-4-yl)ethylamino]-6-phenylpyridazine (3y), which shows an IC50 of 0.12 microM on purified AChE (electric eel), was found to be one of the most potent anti-AChE inhibitors, representing a 5000-fold increase in potency compared to minaprine.1

Emotion regulation as a mediator of associations between mother-child attachment and peer relationships in middle childhood.

Although a link between attachment and peer relationships has been established, the mechanisms that account for this link have not been identified. The 1st goal of this study was to test emotion regulation as a mediator of this link in middle childhood. The 2nd goal was to examine how different aspects of emotion regulation relate to peer competence. Fifth graders completed self-report and semiprojective measures to index mother-child attachment, mothers reported on children's emotionality and coping strategies, and teachers reported on children's peer competence. Constructive coping was related to both attachment and peer competence, and mediated the association between attachment and peer competence, suggesting that emotion regulation is one of the mechanisms accounting for attachment-peer links. Constructive coping was more strongly associated with peer competence for children high on negative emotionality than for children low on negative emotionality.

Dissociation as a mediator between child abuse history and adult abuse potential.

OBJECTIVE: It has commonly been found that abused children are at risk for later becoming abusive parents (Kaufman & Zigler. 1987; Oliver, 1993) and observational learning has been discussed as a mechanism that perpetuates this intergenerational cycle of abuse. However, two thirds of abused children do not become abusive (Kaufman & Zigler, 1987). Thus, the goal of the current study was to examine whether dissociation functions as an additional mechanism mediating the relation between a history of child abuse and abusiveness as an adult. METHOD: A cross-sectional design was used to examine physical abuse history, dissociation, and physical abuse potential in a sample of 190 college students. Questionnaires were utilized to assess the three constructs. RESULTS: Findings indicated that the three constructs were intercorrelated. In particular, a newly established association was found between dissociation and physical abuse potential (r = .54, p < .0001). The primary finding was that the relation between physical abuse history and physical abuse potential was significantly mediated (z = 2.19, p < .05) by level of dissociation, with dissociation accounting for approximately half of the observed relation between history of abuse and abuse potential. CONCLUSION: Results suggested that dissociation may be one mechanism that helps to perpetuate the intergenerational cycle of abuse. Although dissociation promotes psychological survival during children's abuse experiences, it may result in the development of abusive tendencies in later life. Reducing parental dissociation may assist clinicians in preventing or terminating physical child abuse.

Compliance with dietary and nutrient recommendations in the European Prospective Investigation into Cancer and Nutrition (EPIC)-Granada cohort at recruitment.

BACKGROUND: The overall intake of energy and nutrients in the Granada EPIC-cohort (European Prospective Investigation into Cancer and Nutrition) is examined in order to assess compliance with the Spanish Nutritional Objectives (NO) and the Recommended Intakes (RI). METHODS: During recruitment (1992-1996), 7,789 participants, aged 35-69, were asked about diet through a validated diet history questionnaire. Nutrient intake is compared to the NO and RI that were valid at that time. Risk of inadequate intake is estimated as the percentage of the sample with intakes: ≤ 1/3 RI (high risk), ≤ 2/3 RI- > 1/3 RI (moderate risk), ≤ RI- > 2/3 RI, > RI. Differences in intakes have been analyzed by sex and age, and by smoking status and BMI. RESULTS: The daily intake of nutrients did not meet the NO as the total contribution of energy from proteins and fats exceeded these guidelines. Whilst intake of most nutrients was above the RI, the amount of iron, magnesium and vitamins D and E provided by the diet was not enough to meet the RI: in women aged 20-49 years, about 55% were at moderate risk for iron inadequacy, and a 20% of women for magnesium. Both sexes were at high risk of inadequacy for vitamin D, although sunlight exposure may supply adequate amounts. Never smokers showed a higher compliance to the NO. CONCLUSION: At recruitment, the nutrient profile of the diet was unbalanced. The observed nutrient inadequacy for iron, magnesium and vitamin E might be attributed to inappropriate dietary habits, and may have implications for future disease risk.

Natural mentor relationships among Latina adolescent mothers: psychological adjustment, moderating processes, and the role of early parental acceptance.

Investigated the antecedents, effects, and underlying characteristics of natural mentor relationships in a sample of 54 inner-city, Latina adolescent mothers. Women with mentors reported significantly lower levels of depression and anxiety than those without mentors despite similar levels of stress exposure and overall support resources. Young women with natural mentors were also more satisfied with their support resources and appeared better able to cope effectively with relationship problems. Finally, women with mentors recalled their childhood relationships with their mothers as more accepting. Mentor relationships appear to enhance young women's capacity to benefit from their support resources and offset the effects of relationship problems. Implications for future research and intervention strategies are discussed.

Structure-based 3D QSAR and design of novel acetylcholinesterase inhibitors.

The paper describes the construction, validation and application of a structure-based 3D QSAR model of novel acetylcholinesterase (AChE) inhibitors. Initial use was made of four X-ray structures of AChE complexed with small, non-specific inhibitors to create a model of the binding of recently developed aminopyridazine derivatives. Combined automated and manual docking methods were applied to dock the co-crystallized inhibitors into the binding pocket. Validation of the modelling process was achieved by comparing the predicted enzyme-bound conformation with the known conformation in the X-ray structure. The successful prediction of the binding conformation of the known inhibitors gave confidence that we could use our model to evaluate the binding conformation of the aminopyridazine compounds. The alignment of 42 aminopyridazine compounds derived by the docking procedure was taken as the basis for a 3D QSAR analysis applying the GRID/GOLPE method. A model of high quality was obtained using the GRID water probe, as confirmed by the cross-validation method (q2LOO = 0.937, q2L50%O = 0.910). The validated model, together with the information obtained from the calculated AChE-inhibitor complexes, were considered for the design of novel compounds. Seven designed inhibitors which were synthesized and tested were shown to be highly active. After performing our modelling study the X-ray structure of AChE complexed with donepezil, an inhibitor structurally related to the developed aminopyirdazines, has been made available. The good agreement found between the predicted binding conformation of the aminopyridazines and the one observed for donepezil in the crystal structure further supports our developed model.

[Study of sinus node function and atrioventricular conduction in patients with chagas disease (author's transl)]. / Estudio de la función del nodo sinusal y de la conducción atrioventricular en pacientes con enfermedad de chagas.

In order to detect possible abnormalities in sinus node function and atrio-ventricular conduction, one hundred and fourty three chagasic patients were subjected to electrophysiologic and pharmacologic studies. These patients were placed in four categories based on the their clinical, hemodynamic and angiographic characteristics. The chagasic patient without cardiac involvement (group IA), had no detectable abnormalities. Ten percent of those patients with early myocardial damage (group IB), had impaired sinus node automaticity. In three percent, the autonomic innervation was also altered and four percent had abnormally prolonged atrio-ventricular conduction. Among chagasic patients with abnormal EKG'S without heart failure (Group II), 45% had impaired sinus node automaticity, autonomic innervation was altered in 12%, and atrio-ventricular conduction was prolonged in 37%. Sinus node automaticity was impaired in 22% of those patients with abnormal EKG'S and heart failure (Group III). Autonomic innervation was altered in 33%, and atrio-ventricular conduction was prolonged in 47%. We conclude that over half of our patients with abnormal EKG'S will eventually require permanent pacing. Therefore, in these chagasic patients sinus node function and atrioventricular conduction should be routinely studied.

[Echocardiographic evaluation of pulmonary arterial hypertension]. / Evaluación ecocardiográfica de la hipertensión arterial pulmonar.

We performed an echocardiographic and hemodynamic study in 10 patients with cath proven pulmonary arterial hypertension (PAH). The results show that the echocardiographic pattern of PTH such as flat diastolic pulmonary echo, premature closing, flutter, abscense of "A" wave, in creased opening velocity and changes in the right ventricular systolic time intervals, are equivocal. The most common echo finding in our study, was the increased opening velocity. Other echo data could be absent despite the true presence of pulmonary hypertension in a particular case. This report shows similar findings to a recent one published elsewhere (12) with no correlation between echo and hemodynamic data.

[Evaluation of acute effect of GnRH administration on leptin secretion in normal and hyperandrogenic women]. / Evaluación del efecto agudo de la administración de GnRH sobre la secreción de leptina en mujeres sanas e hiperandrogénicas.

BACKGROUND: Several studies suggest that leptin modulates the reproductive axis function. Leptin may stimulate release of GnRH from hypothalamus and of gonadotrophins from the pituitary. A synchronicity of LH and leptin pulses has been described in healthy women and in patients with polycystic ovarian syndrome (PCOS), suggesting a relationship between the episodic secretion of LH and leptin. In vitro experimental studies have demonstrated that leptin administration promotes GnRH-LH release. However it is not established whether GnRH promotes the episodic secretion of leptin. AIM: To assess the response of LH and leptin to the administration of a GnRH bolus in hyperandrogenic and healthy women. PATIENTS AND METHODS: Eleven hyperandrogenic and eleven healthy women of similar age and body mass index (BMI) were studied. Under basal conditions three blood samples were collected every 30 min before and after the administration of a GnRH bolus (100 micrograms). LH and leptin concentrations were measured in all samples. Testosterone, SHBG and estradiol were determined in the first sample. For data analysis, the increment of LH and leptin between 0-30 and 0-60 min was calculated. The LH and leptin areas under the curve (AUC) before and after GnRH administration were also calculated in both groups. RESULTS: After GnRH administration an increment in LH concentrations was observed in both groups; however, leptin concentrations were not modified. In both groups LH area under the curve increased after GnRH administration; however, the leptin area was not modified. CONCLUSIONS: These results suggest that circulating leptin concentration is not modulated by GnRH-LH.