Potencial energético de la biomasa procedente de árboles frutales podados en Croacia

 The world's most developed countries and the European Union (EU) deem that the renewable energy sources should partly substitute fossil fuels and become a “bridge” to the utilization of other energy sources of the future. This paper will present the possibility of using pruned biomass from fruit cultivars. It will also present the calculation of potential energy from the mentioned raw materials in order to determine the extent of replacement of non-renewable sources with these types of renewable energy. One of the results of the intensive fruit-growing process, in post pruning stage, is large amount of pruned biomass waste. Based on the calculated biomass (kg ha-1) from intensively grown woody fruit crops that are most grown in Croatia (apple, pear, apricots, peach and nectarine, sweet cherry, sour cherry, prune, walnut, hazelnut, almond, fig, grapevine, and olive) and the analysis of combustible (carbon 45.55-49.28%, hydrogen 5.91-6.83%, and sulphur 0.18-0.21%) and non-combustible matters (oxygen 43.34-46.6%, nitrogen 0.54-1.05%, moisture 3.65-8.83%, ashes 1.52-5.39%) with impact of lowering the biomass heating value (15.602-17.727 MJ kg-1), the energy potential of the pruned fruit biomass is calculated at 4.21 PJ.Los principales países desarrollados del mundo y de la Unión Europea (UE) consideran que las fuentes de energía renovables deberían sustituir parcialmente a los combustibles fósiles y convertirse en el futuro en un “puente” hacia la utilización de otras fuentes de energía. En este trabajo, se planteó la posibilidad de utilizar biomasa cortada procedente de cultivos frutales con el propósito de calcular el potencial energético del mencionado material en bruto, así como determinar el grado de remplazo de fuentes no renovables con este tipo de fuentes de energía renovable. Uno de los resultados del proceso de cultivo de frutales en intensivo, tras la época de poda, es la gran cantidad de biomasa cortada inútil. En base al cálculo de biomasa (kg ha-1) de los frutales leñosos de cultivo intensivo más comunes en el territorio de Croacia (manzano, peral, albaricoquero, melocotonero y nectarino, cerezo, guindo, ciruelo, nogal, avellano, almendro, higuera, viña y olivo) y en base al análisis de partículas combustibles (carbono 45,55-49,28%, hidrógeno 5,91-6,83% y azufre 0,18-0,21%) y de partículas no combustibles (oxígeno 43,34-46,6%, nitrógeno 0,54-1,05%, vapor de agua 3,65-8,83% y cenizas 1,52-5,39%) que influyen en el poder calorífico  inferior de la biomasa (15,602-17,727 MJ kg-1), se calcula que la energía potencial de los restos de poda de frutales es 4.21 PJ

Hybridization Assays Using an Expressible DNA Fragment Encoding Firefly Luciferase as a Label

We report the use of a new label, an expressible enzymecoding DNA fragment, for nucleic acid hybridization assays. The DNA label contains a firefly luciferase coding sequence downstream from a T7 RNA polymerase promoter. The target DNA (200 bp) is denatured and hybridized simultaneously with two oligonucleotide probes. One of the probes is immobilized in microtiter wells, via the digoxigenin/anti-digoxigenin interaction, and the other probe is biotinylated. After completion of the hybridization, the hybrids are reacted with a streptavidin-luciferase DNA complex. Subsequently, the solid-phase bound DNA is expressed by coupled transcription/ translation. The synthesized luciferase catalyzes the luminescent reaction of luciferin with O 2 and ATP. The luminescence is linearly related to the amount of target DNA in the range of 5-5000 amol. The CVs obtained for 20 and 100 amol of target are 6.5% and 10.8%, respectively (n ) 4). The specific and strong interaction between two complementary nucleic acid strands forms the basis for the development of hybridization assays. Hybridization methodology is emerging as the most promising area in laboratory medicine and has transformed the way clinical testing is realized. Previous tests have been based on the monitoring of gene products, i.e., phenotypic markers, such as oncoproteins, viral antigens, etc. In contrast, current laboratory tests that are based on hybridization allow the analysis of disease at the nucleic acid level. Thus, pre-or postnatal diagnosis of genetic disease can be accomplished by hybridization of the patient's DNA with allele-specific oligonucleotide probes that recognize mutations, deletions, or insertions causing the disease. Also, the various infectious agents can be measured in biological fluids by hybridization with specific probes. In forensic science, hybridization of DNA with minisatellite probes allows the unique identification of individuals (DNA fingerprinting). 1,2 Radioactive probes (usually labeled with 32 P), in combination with autoradiographic detection, dominated in the field of hybridization assays for more than 2 decades and provide the highest sensitivities. However, the short half-life of 32 P, the health hazards and problems associated with its use and disposal, and the long exposure times (many hours to days) required for detection have placed limitations on the routine use of hybridization assays in the clinical laboratory. The current trend in this area is toward novel nonradioactive alternatives. 2,3 The labels can be incorporated into the probes either enzymatically (e.g., using DNA polymerase or deoxynucleotidyl transferase and modified deoxynucleoside triphosphates) or by chemical conjugation (e.g., introduction of NH 2 groups into the probe via cytidine transamination and then conjugation to the reporter molecule). 4 Nonisotopic hybridization assays based on fluorescent, chemiluminescent, or enzyme labels have been developed. Generally, there are two strategies for the analysis of hybrids. Either the reporter molecule is directly conjugated to the probe, 5,6 or a ligand is attached to the probe and the hybrids are measured in a subsequent step by adding a specific, labeled binding protein. The ligand may be biotin or a hapten (e.g., digoxigenin). Labeled (strept)avidin or antihapten antibodies may then be employed for detection. 7,8 Enzymes (such as alkaline phosphatase and horseradish peroxidase) are the most widely used nonradioisotopic labels because they provide amplification through the high turnover of their substrates to detectable products. 2 Recently, we reported 9 that a DNA fragment (DNA template) coding for an enzyme can be used as a novel label for the development of highly sensitive immunoassays (expression immunoassays). In these assays, after completion of the immunoreaction, the DNA template (a luciferase-coding DNA) is expressed by in vitro transcription/translation and the activity of the synthesized enzyme is measured. Furthermore, it was estimated that 12-14 luciferase molecules were synthesized from each DNA template molecule. In the present work, we extend our investigation in the area of hybridization assays. EXPERIMENTAL SECTION Instrumentation. Luminescence measurements were carried out using a liquid scintillation counter (Model LS-6500, Beckman Instruments Inc., Fullerton, CA) in the single photon monitoring mode. Fluorescence measurements were performed with th

In vitro metabolism of beclomethasone dipropionate, budesonide, ciclesonide, and fluticasone propionate in human lung precision-cut tissue slices

<p>Abstract</p> <p>Background</p> <p>The therapeutic effect of inhaled corticosteroids (ICS) may be affected by the metabolism of the drug in the target organ. We investigated the <it>in vitro </it>metabolism of beclomethasone dipropionate (BDP), budesonide (BUD), ciclesonide (CIC), and fluticasone propionate (FP) in human lung precision-cut tissue slices. CIC, a new generation ICS, is hydrolyzed by esterases in the upper and lower airways to its pharmacologically active metabolite desisobutyryl-ciclesonide (des-CIC).</p> <p>Methods</p> <p>Lung tissue slices were incubated with BDP, BUD, CIC, and FP (initial target concentration of 25 μM) for 2, 6, and 24 h. Cellular viability was assessed using adenosine 5'-triphosphate content and protein synthesis in lung slices. Metabolites and remaining parent compounds in the tissue samples were analyzed by HPLC with UV detection.</p> <p>Results</p> <p>BDP was hydrolyzed to the pharmacologically active metabolite beclomethasone-17-monopropionate (BMP) and, predominantly, to inactive beclomethasone (BOH). CIC was hydrolyzed initially to des-CIC with a slower rate compared to BDP. A distinctly smaller amount (approximately 10-fold less) of fatty acid esters were formed by BMP (and/or BOH) than by BUD or des-CIC. The highest relative amounts of fatty acid esters were detected for BUD. For FP, no metabolites were detected at any time point. The amount of drug-related material in lung tissue (based on initial concentrations) at 24 h was highest for CIC, followed by BUD and FP; the smallest amount was detected for BDP.</p> <p>Conclusion</p> <p>The <it>in vitro </it>metabolic pathways of the tested ICS in human lung tissue were differing. While FP was metabolically stable, the majority of BDP was converted to inactive polar metabolites. The formation of fatty acid conjugates was confirmed for BMP (and/or BOH), BUD, and des-CIC.</p

Linoleic Acid-Induced Ultra-Weak Photon Emission from Chlamydomonas reinhardtii as a Tool for Monitoring of Lipid Peroxidation in the Cell Membranes

Reactive oxygen species formed as a response to various abiotic and biotic stresses cause an oxidative damage of cellular component such are lipids, proteins and nucleic acids. Lipid peroxidation is considered as one of the major processes responsible for the oxidative damage of the polyunsaturated fatty acid in the cell membranes. Various methods such as a loss of polyunsaturated fatty acids, amount of the primary and the secondary products are used to monitor the level of lipid peroxidation. To investigate the use of ultra-weak photon emission as a non-invasive tool for monitoring of lipid peroxidation, the involvement of lipid peroxidation in ultra-weak photon emission was studied in the unicellular green alga Chlamydomonas reinhardtii. Lipid peroxidation initiated by addition of exogenous linoleic acid to the cells was monitored by ultra-weak photon emission measured with the employment of highly sensitive charged couple device camera and photomultiplier tube. It was found that the addition of linoleic acid to the cells significantly increased the ultra-weak photon emission that correlates with the accumulation of lipid peroxidation product as measured using thiobarbituric acid assay. Scavenging of hydroxyl radical by mannitol, inhibition of intrinsic lipoxygenase by catechol and removal of molecular oxygen considerably suppressed ultra-weak photon emission measured after the addition of linoleic acid. The photon emission dominated at the red region of the spectrum with emission maximum at 680 nm. These observations reveal that the oxidation of linoleic acid by hydroxyl radical and intrinsic lipoxygenase results in the ultra-weak photon emission. Electronically excited species such as excited triplet carbonyls are the likely candidates for the primary excited species formed during the lipid peroxidation, whereas chlorophylls are the final emitters of photons. We propose here that the ultra-weak photon emission can be used as a non-invasive tool for the detection of lipid peroxidation in the cell membranes

Epidemiology of intra-abdominal infection and sepsis in critically ill patients: “AbSeS”, a multinational observational cohort study and ESICM Trials Group Project

Purpose: To describe the epidemiology of intra-abdominal infection in an international cohort of ICU patients according to a new system that classifies cases according to setting of infection acquisition (community-acquired, early onset hospital-acquired, and late-onset hospital-acquired), anatomical disruption (absent or present with localized or diffuse peritonitis), and severity of disease expression (infection, sepsis, and septic shock). Methods: We performed a multicenter (n = 309), observational, epidemiological study including adult ICU patients diagnosed with intra-abdominal infection. Risk factors for mortality were assessed by logistic regression analysis. Results: The cohort included 2621 patients. Setting of infection acquisition was community-acquired in 31.6%, early onset hospital-acquired in 25%, and late-onset hospital-acquired in 43.4% of patients. Overall prevalence of antimicrobial resistance was 26.3% and difficult-to-treat resistant Gram-negative bacteria 4.3%, with great variation according to geographic region. No difference in prevalence of antimicrobial resistance was observed according to setting of infection acquisition. Overall mortality was 29.1%. Independent risk factors for mortality included late-onset hospital-acquired infection, diffuse peritonitis, sepsis, septic shock, older age, malnutrition, liver failure, congestive heart failure, antimicrobial resistance (either methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, extended-spectrum beta-lactamase-producing Gram-negative bacteria, or carbapenem-resistant Gram-negative bacteria) and source control failure evidenced by either the need for surgical revision or persistent inflammation. Conclusion: This multinational, heterogeneous cohort of ICU patients with intra-abdominal infection revealed that setting of infection acquisition, anatomical disruption, and severity of disease expression are disease-specific phenotypic characteristics associated with outcome, irrespective of the type of infection. Antimicrobial resistance is equally common in community-acquired as in hospital-acquired infection