Search

Calcitonin gene-related peptide (CGRP), vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating polypetide-38 (PACAP-38) have relevant roles in migraine pathophysiology. Their serum levels have been proposed as biomarkers for migraine. Our aim was to assess their diagnostic value in real clinical practice in a cohort of chronic migraine (CM), episodic migraine (EM) and healthy controls (HC). We recruited subjects with CM, EM and HC at two medical centers. Blood samples were drawn under fasting conditions in the interictal period, immediately centrifuged and stored at - 80 ºC. Serum levels were determined by ELISA. Neuropeptide levels, the effect of preventatives, correlations with clinical and demographic variables, and their diagnostic value were studied among clinical categories. 296 age- and sex-matched subjects (101 CM, 98 EM and 97 HC) were included. All three neuropeptide serum levels were higher in CM [median and IQ for CGRP = 18.023 pg/ml (14.4-24.7); VIP = 121.732 pg/ml (48.72-186.72) and PACAP = 204.931 pg/ml (101.08-597.64)] vs EM [CGRP = 14.659 pg/ml (10.29-17.45); VIP = 75.603 pg/ml (28.722-107.10); and PACAP = 94.992 pg/ml (65.77-128.48)] and vs HC [CGRP = 13.988 pg/ml (10.095-17.87); VIP = 84.685 pg/ml (35.32-99.79), and PACAP = 103.142 pg/ml (59.42-123.97)]. Using multinomial modeling, only VIP (OR 1.011, 95% CI  1.003-1.018, p = 0.005) and PACAP (OR 1.003, 95% CI 1.001-1.005, p = 0.002) increased the risk for CM, but not for EM. CGRP did not predict CM or EM. This model could correctly classify only 62/101 (61.38%) of CM, 75/98 (76.53%) of EM, and 5/97 (4.12%) of HC [globally 147/296 (49.8%)]. Individually, PACAP performed the best for classifying clinical categories [global accuracy 150/296 (50.67%)]. In CM, neuropeptide levels were higher in those OnaBT-treated than in no-treated patients. Although interictal serum CGRP and VIP were higher in CM than both EM or HC, their utility to discriminate migraine categories was low. Contrary to other studies, PACAP serum levels were also higher in CM than in EM or HC and had more discriminative capability to distinguish CM from EM and HC. Further investigation is needed for determination technique standardization.

Optimal foraging theory has typically paid little attention to species feeding on mobile prey and has emphasised energy intake rather than the nutritional contribution of food. The difficulty of capturing food has rarely been included in foraging models, even when it is a potentially important modulator of time devoted to foraging. From the central place foraging and provisioning perspectives, it is posited that at high levels of prey selectivity, the time spent to capture prey is longer than at low levels of prey selectivity. Furthermore, in the case of carnivorous predators, it is thought that nutritional composition does not influence foraging strategies. To explore these issues, we investigated the influence of abundance, size, difficulty of capture, gross energy and nutritional composition (fat, protein, protein-fat ratio and amino acid contents) of prey species on the foraging behaviour of a predator species, the common kestrel Falco tinnunculus, in a region of high diversity of prey species. Our results show that capturability index and load-size explain the foraging behaviour of kestrels. Preferred prey take longer to be provisioned, both selectivity and capturability might explain this result. It is also shown that specific nutritional components, such as protein and amino acid contents, are likely to explain food preference in this carnivorous-insectivorous species.

Sea hares of Aplysia genus are recognized as a source of a diverse range of metabolites. 5α,8α-Endoperoxides belong to a group of oxidized sterols commonly found in marine organisms and display several bioactivities, including antimicrobial, anti-tumor, and immunomodulatory properties. Herein we report the isolation of 5α,8α-epidioxycholest-6-en-3β-ol (EnP(5,8)) from Aplysia depilans Gmelin, based on bioguided fractionation and nuclear magnetic resonance (NMR) analysis, as well as the first disclosure of its anti-inflammatory properties. EnP(5,8) revealed capacity to decrease cellular nitric oxide (NO) levels in RAW 264.7 macrophages treated with lipopolysaccharide (LPS) by downregulation of the Nos2 (inducible nitric oxide synthase, iNOS) gene. Moreover, EnP(5,8) also inhibited the LPS-induced expression of cyclooxygenase-2 (COX-2), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) at the mRNA and protein levels. Mild selective inhibition of COX-2 enzyme activity was also evidenced. Our findings provide evidence of EnP(5,8) as a potential lead drug molecule for the development of new anti-inflammatory agents.

Background: The goal of this study was to assess the effect of associated gastrointestinal malformations (GI) on the outcomes of patients undergoing congenital heart operations., Methods: Neonates and infants with thoracic (esophageal atresia, tracheoesophageal fistula) and abdominal (duodenal stenosis/atresia, imperforate anus, Hirschsprung disease) GI malformations undergoing congenital heart operations between 1995 and 2015 were included. Two control groups were created, one for each group. Patients were matched by diagnosis, procedure, history of prematurity, presence of genetic syndrome, and a propensity score including weight and year of operation., Results: The cohort included 383 patients: 52 (14%) with thoracic GI malformations and 98 (25%) thoracic GI controls, 80 (21%) with abdominal GI malformations and 153 (40%) abdominal GI controls. Median follow-up was 6 years (range, 16 days to 20 years). Patients with thoracic GI malformations had longer length of stay (p < 0.001), longer intubation times (p = 0.002), and higher perioperative death (p = 0.015) than controls. There was a tendency for worse overall survival than controls, mainly explained by the higher risk of early death (p = 0.06). No difference was found in outcomes between patients with abdominal GI malformations and controls., Conclusions: Patients with thoracic GI malformations have worse perioperative outcomes than controls, but their long-term survival does not seem to be significantly different. Abdominal GI malformations do not have a significant effect on outcomes. The presence of GI malformations should likely not preclude patients from undergoing congenital heart operations, but careful family counseling is necessary, especially for thoracic GI malformations., (Copyright © 2017 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

In a rat model of right free wall replacement, the transplantation of an engineered multilayered myocardial patch fabricated from a polycaprolactone membrane supporting a chitosan/heart matrix hydrogel induces significant muscular and vascular remodeling and results in a significantly higher right ventricular ejection fraction compared to use of a commercially available pericardium patch., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Predictors for single ventricle palliation (SVP) or successful biventricular repair (BVR) in patients with borderline left-side heart structures are not well defined. The goal was to evaluate the role of echocardiography and intracardiac exploration in determining feasibility of BVR., Methods: All neonates surgically treated from 1995 to 2015 with mitral valve (MV), aortic valve, or left ventricle end-diastolic dimension z score of -2 or less for whom management was controversial were included. Data were analyzed using Fisher's exact test, Kruskal-Wallis test, and Kaplan-Meier analysis., Results: The cohort consisted of 42 patients: 7 SVP (17%) and 35 BVR (83%). Median follow-up was 7 years (range, 6 months to 18 years). Intracardiac exploration was performed in 29 patients (69%). There was poor correlation between echocardiographic and intraoperative MV measurements (intraclass correlation coefficient 0.14). Preoperative echocardiography significantly underestimated MV size in 14 patients (54%). Two BVR patients were converted to SVP, and 4 (including 1 converted patient) had cardiac-related deaths. All patients with MV greater than 8 mm on preoperative echocardiography had successful BVR. An intraoperative MV less than 8 mm and an abnormal subvalvar apparatus was present in 5 of 6 SVP (83%) and 3 of 3 (100%) failed BVR patients who had intracardiac exploration, and in only 1 of 20 successful BVR patients (5%) who had an intracardiac exploration., Conclusions: The decision to proceed to BVR in patients with borderline left-side heart structures should not rely strictly on echocardiographic measurements. Intracardiac exploration of the MV and subvalvar apparatus is useful before committing a patient to SVP. Patients with low MV z scores, especially those with a normal subvalvar apparatus, may undergo BVR with good outcomes., (Copyright © 2017 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Objective: Routine preoperative laboratory testing (RLT) is common practice in pediatric cardiothoracic surgery and is associated with significant cost burden to patients and families. We sought to examine the value of RLT in patients undergoing elective pediatric cardiothoracic surgery., Methods: We conducted a retrospective study of all scheduled elective pediatric cardiothoracic surgery at our institution from 2012 to 2014. Inpatients were excluded. Patient charts were reviewed to obtain preoperative laboratory values and determine relationship to case cancellation. RLT includes complete blood count, prothrombin time, partial thromboplastin time, urinalysis, 7 chemistry metabolic panel, electrocardiogram, and 2-view chest radiograph., Results: RLT was completed for 1106 scheduled elective cases. Six (0.5%) cancellations were related to abnormal preoperative laboratory test results: 5 complete blood counts and 1 urinalysis. Hospital charge for RLT averaged $2064 per patient. Based on this incidence, we estimated that 184 routine preoperative laboratory tests, which generated a total hospital charge of $379,776, were required to capture 1 abnormal test significant enough to cancel surgery. An estimated charge of $2,169,552 was generated on prothrombin time, partial thromboplastin time, 7 chemistry metabolic panel, electrocardiogram, and 2-view chest radiograph, and none of these tests resulted in a cancellation., Conclusions: RLT does not significantly impact decision-making in elective pediatric cardiothoracic surgery. The decision to order a specific screening test should be clinically driven. Selective preoperative laboratory testing may have a positive impact on healthcare costs without affecting outcomes., (Copyright © 2016. Published by Elsevier Inc.)

The stiffness of myocardial tissue changes significantly at birth and during neonatal development, concurrent with significant changes in contractile and electrical maturation of cardiomyocytes. Previous studies by our group have shown that cardiomyocytes generate maximum contractile force when cultured on a substrate with a stiffness approximating native cardiac tissue. However, effects of substrate stiffness on the electrophysiology and ion currents in cardiomyocytes have not been fully characterized. In this study, neonatal rat ventricular myocytes were cultured on the surface of flat polyacrylamide hydrogels with elastic moduli ranging from 1 to 25 kPa. Using whole-cell patch clamping, action potentials and L-type calcium currents were recorded. Cardiomyocytes cultured on hydrogels with a 9 kPa elastic modulus, similar to that of native myocardium, had the longest action potential duration. Additionally, the voltage at maximum calcium flux significantly decreased in cardiomyocytes on hydrogels with an elastic modulus higher than 9 kPa, and the mean inactivation voltage decreased with increasing stiffness. Interestingly, the expression of the L-type calcium channel subunit α gene and channel localization did not change with stiffness. Substrate stiffness significantly affects action potential length and calcium flux in cultured neonatal rat cardiomyocytes in a manner that may be unrelated to calcium channel expression. These results may explain functional differences in cardiomyocytes resulting from changes in the elastic modulus of the extracellular matrix, as observed during embryonic development, in ischemic regions of the heart after myocardial infarction, and during dilated cardiomyopathy.

Taking into account the broad biological activities found in the meridianin indole alkaloids isolated to date, we have re-examined the organic extracts of an Antarctic collection of the tunicates Aplidium meridianum and A. falklandicum (Chordata: Ascidiacea) by HPLC in conjunction with a high-resolution mass spectrometer (HPLC-MS). A new set of analogs of meridianins A-G has been detected, and their structures are proposed on the basis of the molecular formulae identified by LC-HRMS analysis using a C18 column with a gradient of water/acetonitrile and an LTQ-FT-MS Orbitrap detector. Remarkably, dimers derived from meridianin A and from meridianin B or E were also detected. Our findings provide further evidence of the broad variability within the meridianin-like derivatives of this highly bioactive alkaloid family., (Copyright © 2015 John Wiley & Sons, Ltd.)

Thelepamide (1) was characterized during a program to study cytotoxic substances from an unusual source, the tidal zone-derived annelid Thelephus crispus. Its structure contains a tetraketide and a tripeptide subunit and possesses striking atom diversity, consisting of 17 carbons and 8 heteroatoms. The relative configurations at four chiral sites were elucidated via ROESY, J-based configurational analysis, and DFT calculations. It was modestly active against leukemia cells (IC50 = 5 μg/mL) and inactive against solid tumor cell lines.

Ascidians have developed multiple defensive strategies mostly related to physical, nutritional or chemical properties of the tunic. One of such is chemical defense based on secondary metabolites. We analyzed a series of colonial Antarctic ascidians from deep-water collections belonging to the genera Aplidium and Synoicum to evaluate the incidence of organic deterrents and their variability. The ether fractions from 15 samples including specimens of the species A. falklandicum, A. fuegiense, A. meridianum, A. millari and S. adareanum were subjected to feeding assays towards two relevant sympatric predators: the starfish Odontaster validus, and the amphipod Cheirimedon femoratus. All samples revealed repellency. Nonetheless, some colonies concentrated defensive chemicals in internal body-regions rather than in the tunic. Four ascidian-derived meroterpenoids, rossinones B and the three derivatives 2,3-epoxy-rossinone B, 3-epi-rossinone B, 5,6-epoxy-rossinone B, and the indole alkaloids meridianins A-G, along with other minoritary meridianin compounds were isolated from several samples. Some purified metabolites were tested in feeding assays exhibiting potent unpalatabilities, thus revealing their role in predation avoidance. Ascidian extracts and purified compound-fractions were further assessed in antibacterial tests against a marine Antarctic bacterium. Only the meridianins showed inhibition activity, demonstrating a multifunctional defensive role. According to their occurrence in nature and within our colonial specimens, the possible origin of both types of metabolites is discussed.

Purpose: To analyze the impact of storage temperature on the 2D-DIGE profile of human plasma., Experimental Design: 2D-DIGE and MS were used to identify the differences in the proteomic profiles of aliquots of eight human plasma samples stored at either - 30 or - 80°C for 18 months., Results: After the depletion of albumin and IgG, 2D-DIGE identified four spots significantly and reproducibly increased, and five spots that were decreased, in samples stored at - 30°C compared with those stored at - 80°C. These nine spots were manually excised, digested ingel and analyzed by MALDI-TOF/TOF. MASCOT database search using the PMF spectra allowed the identification of the proteins present in eight of the nine spots. All the spots corresponded to the complement C3 precursor protein., Conclusions and Clinical Relevance: Our present results indicate that plasma collections stored at -30°C, and not only those stored at - 80°C, may be used for 2D-DIGE analyses of albumin-and IgG-depleted human plasma without loosing the essential information about highly abundant proteins. This finding expands the applicability of 2D-DIGE to the study of human disease by permitting the analysis of human plasma samples stored at temperatures between - 30 and - 80°C.

The protozoan Entamoeba histolytica is the etiological agent of amebiasis, an infection with high prevalence worldwide. The host-ameba relationship outcome depends on parasite and host factors, and among these is secretory IgA. These antibodies reduce mucosal colonization by pathogens and neutralize a variety of toxins and enzymes. The functionality of secretory IgA depends on its integrity. Some bacteria produce IgA proteases that cleave mainly the IgA1 subclass; live E. histolytica trophozoites, and other ameba fractions are also able to degrade human IgA. The aim of this study was to determine if serum and secretory IgA, its subclasses and secretory component, are degraded by cysteine proteases, which are present and active on the surface of glutaraldehyde-fixed amebas. It was observed that secretory IgA1, IgA2, free and IgA-bound secretory component were degraded by E. histolytica surface-associated cysteine proteinases. Secretory IgA2, although it was degraded, conserved its ability to agglutinate live amebas better than IgA1. Therefore, while specificity of known ameba cysteine proteases is cathepsin B-like and is different from bacterial IgA proteases, IgA2 was functionally more resistant than IgA1 to ameba surface-associated cysteine protease degradation, similar to the greater resistance of IgA2 to bacterial IgA-specific proteases.

Objective: To compare postnatal growth and nutritional deficits after the implementation of two different nutritional strategies in two consecutives periods of time., Methods: An early and aggressive nutritional regimen was used in a cohort of 117 very low birth weight (VLBW) infants. Amino acids were administered at the rate of 1.5 g/kg/day along with 5.6 mg/k/min of glucose flow on day 1 of life, and progressively increased to 4 g/kg/day and 13 mg/kg/min. Intravenous lipids were started at 0.5 g/kg/day at 24 h from birth, and increased to 3.5 g/kg/day; enteral feeding was begun at day 1 of life. Uni- and multivariate analyses were used to compare this group with the conventional group of 65 VLBW infants conservatively fed., Results: Univariate analysis showed that in the aggressive group there was a 66% reduction in the risk of post natal malnutrition at 40 weeks of postmenstrual age (OR 0.34; 95% CI 0.17-0.67). This difference persisted in the multivariate analysis. Energy and protein deficits were lower in the aggressive group (P < 0.001)., Conclusions: Early and aggressive introduction of total parenteral nutrition and enteral feeding resulted in better growth in weight, length and head circumference, and a reduction of nutritional deficits at 40 weeks of postmenstrual age.

A series of 6-methyl-1H-pyrimidin-2,4-diones bearing different substituents in the 1-position of the uracil ring were prepared starting from substituted ureas and diketene.

We have identified in plasma membrane fractions isolated from rat hepatocarcinoma AS-30D ascites cells three glycoproteins of 125 kDa, 115 kDa and 105 kDa (gp125, gp115 and gp105) which become adenylylated using ATP as substrate, most readily in the presence of EDTA. The gp115 becomes also phosphorylated. The adenylylation of these tumor glycoproteins was much lower than that of a group of analogous adenylylatable glycoproteins (gp130, gp120-gp110 dimer and gp100) present in normal rat liver plasma membrane. The tumor glycoproteins were reversibly O-adenylylated at threonine residues, as was the case for their normal rat liver counterparts. The tumor gp115, and the gp120-gp110 dimer from normal rat liver were both isolated using either ATP-affinity chromatography and/or AMP-affinity chromatography. The gp120-gp110 dimer from normal rat liver was identified as the plasma cell differentiation antigen-1 (PC-1 protein), an ecto-5' phosphodiesterase/ nucleotide-pyrophosphatase (5'-PDE/NPPase). The gp115 from tumor cells also exhibited Zn2+-stimulated 5'-PDE and NPPase activities in alkaline conditions, although it appears to be distinct from the PC-1 protein. We have determined that the gp115 is an ecto-enzyme that catalyzes the hydrolysis of extracellular ATP, since its adenylylation and phosphorylation were detected in intact cells using extracellularly added [alpha-32P]ATP or [gamma-32P]ATP, respectively, in the absence of any permeabilizing agent.

Phosphocalmodulin has been shown to have a differential biological activity compared to nonphosphorylated calmodulin when assayed on a variety of calmodulin-dependent systems. However, the phosphocalmodulin preparations used so far in those experiments were not necessarily free of nonphosphorylated calmodulin. Therefore, the results obtained may not unquestionably show the real effect of pure phosphocalmodulin on the systems under study. To solve this problem, we describe here a method for the purification of phospho(Tyr)calmodulin free of nonphosphorylated calmodulin. The procedure consists of the following steps: (i) phosphorylation of calmodulin by a fraction enriched in epidermal growth factor receptor tyrosine kinase from rat liver isolated by calmodulin affinity chromatography, (ii) isolation of a calmodulin/phosphocalmodulin mixture by Ca(2+)-dependent chromatography in phenyl-Sepharose, (iii) purification of phospho(Tyr)calmodulin using an anti-phosphotyrosine antibody immobilized in agarose upon elution with phenyl phosphate, and (iv) removal of phenyl phosphate from the phospho(Tyr)calmodulin preparation by filtration chromatography in a Bio-Gel P-2 column. The obtained phospho(Tyr)calmodulin preparation was highly pure and essentially free of nonphosphorylated calmodulin because of the use of anti-phosphotyrosine affinity chromatography. We demonstrate that this ultrapure phospho(Tyr)calmodulin preparation is totally incapable of activating the calmodulin-dependent cyclic nucleotide phosphodiesterase. In contrast, when a nonpurified phospho(Tyr)calmodulin preparation was used a partial activation of this enzyme was observed., (Copyright 1999 Academic Press.)

Weanling rats were offered food ad libitum, or fasted for 18 h, or fasted and refed for times ranging from 5 to 30 min. Five protein kinase C (PKC) isoforms (alpha, epsilon, zeta, theta and mu) were detected in the hindlimb muscles by Western immunoblotting. PKC forms epsilon and theta were abundant in plantaris, but not in soleus muscle, and no difference in localization was detected between fed rats and those fasted for 18 h. PKC forms alpha and mu were affected by fasting and refeeding. PKC-mu was found only in the cytosolic fraction of the plantaris muscle of the fasted animal, but in the fully-fed animals it was also associated with the membrane fraction. The pattern of localization observed in the fully-fed state was restored in the fasted rats by 20 min refeeding. In contrast, PKC-alpha was not detected in the cytosolic fraction of the plantaris in fasted animals but rapidly reappeared there on refeeding, being restored to 20% and 80% of the fed value within 5 and 30 min of refeeding respectively. The timing of these changes was correlated with the increase in serum insulin concentration, which was significantly elevated above the fasted value by 5 min and at subsequent times. These data suggest a possible role for PKC isoforms alpha and mu in the metabolic changes that occur in skeletal muscle on transition between the fasted and the fed state.

The Indonesian gorgonian Briareum sp. has afforded two new briarane stecholide diterpenes: 2,9-diacetyl-2-debutyrylstecholide H (1) and 13-dehydroxystecholide J (2), the semisynthetic 2beta-acetoxy-2-(debutyryloxy)stecholide E acetate (3), which was isolated as a natural product for the first time, along with the known compounds stecholides I-M (4-8), stecholide A acetate (9), and stecholide C acetate (10). The structures of the new compounds were deduced by extensive NMR studies (1H, 13C, 1H-1H COSY, NOESY, HMQC, and HMBC) and FABMS data (HR and LR). Cytotoxic activity was found for the first time in stecholide L (7).

New bengazoles 3-9 as inseparable mixtures are reported from the sponge Jaspis cf. coricea collected in Papua New Guinea. These compounds contain a new variation of the unusual bisoxazole core present in bengazole A [1]. The B-ring oxazole hexatetraol side chain is varyingly substituted with either myristic acid or 13-methylmyristic acid. Hydrolysis of three different bengazole mixtures each yielded an identical tetraol, bengazole Z [11].