Μελέτη της διεργασίας παραγωγής βιοαιθανόλης από διατροφικά απορρίμματα

107 σ.Η παραγωγή βιοκαυσίμων, και συγκεκριμένα βιοαιθανόλης, από βιομάζα αποτελεί το κέντρο επιστημονικού και βιομηχανικού ενδιαφέροντος τα τελευταία χρόνια. Συγκεκριμένα για να αποκτήσει βιώσιμο χαρακτήρα η παραγωγή 2ης γενιάς βιοαιθανόλης, η προσπάθεια έχει επικεντρωθεί στην μείωση του κόστους των πρώτων υλών. Η διαχείριση των διατροφικών απορριμμάτων είναι ένα σημαντικό ζήτημα τόσο για την προστασία του περιβάλλοντος, όσο και για τη διατήρηση των πόρων. Η μετατροπή των υπολειμμάτων των τροφών σε αιθανόλη θα μπορούσε να προσφέρει περαιτέρω οικονομικά πλεονεκτήματα, λόγω της αύξησης του κόστους διάθεσης των απορριμμάτων. Το γεγονός ότι τα διατροφικά απορρίμματα είναι πλούσια σε ζυμώσιμα σάκχαρα, τα καθιστά κατάλληλη πρώτη ύλη για παραγωγή βιοαιθανόλης, ενός εναλλακτικού και ανανεώσιμου καυσίμου κίνησης. Σκοπός της παρούσας διπλωματικής εργασίας είναι η μελέτη της διεργασίας παραγωγής βιοαιθανόλης από διατροφικά απορρίμματα και η βελτιστοποίηση των συνθηκών παραγωγής της. Αρχικά, η πρώτη ύλη προκατεργάζεται υδροθερμικά σε θερμοκρασία 100ο C για 1 hour παρουσία και μη H2SO4 σε περιεκτικότητα 1g/100g στερεού, με συγκέντρωση στερεού 30 % β/ο . Το ποσοστό του αμύλου που αποικοδομείται είναι 47,13% μετά από υδροθερμική προκατεργασία παρουσία H2SO4, 1g/100g ξ.σ. και 45,72% μετά από υδροθερμική προκατεργασία απουσία οξέος. Ακολουθεί ενζυμική υδρόλυση του προκατεργασμένου υλικού με χρήση αμυλολυτικών (Liquozyme SC DS και Spirizyme Fuel ) με ενεργότητες 0,053U/g αμύλου για την α-αμυλάση και 5.06, 10.2, 20.24 U/g αμύλου για την γλυκοαμυλάση και κυτταρινολυτικών (Celluclast 1,5L/Novozyme 188 (5:1) και Cellic® CTec2) εμπορικά διαθέσιμων ενζυμικών σκευασμάτων με ενεργότητες 1,3,5,7,9 FPU/g υλικού. Η υδρόλυση του αμύλου κυμαίνεται μεταξύ 72,30 και 86,32%. Το θεωρητικό ποσοστό υδρόλυσης της κυτταρίνης με βάση την παραχθείσα γλυκόζη κατά την υδρόλυση με μίγμα Celluclast 1,5L/Novozyme 188 (5:1) κυμαίνεται μεταξύ 75,28% και 97,63%, ενώ όσον αφορά στην υδρόλυση με χρήση του σκευάσματος Cellic® CTec2 το αντίστοιχο ποσοστό είναι 25,70% έως 64,22%. Τέλος, πραγματοποιείται μελέτη της συμπεριφοράς των αμυλολυτικών και κυτταρινολυτικών ενζυμικών σκευασμάτων καθώς και της συνεργιστικής τους δράσης κατά την παραγωγή αιθανόλης με εφαρμογή της τεχνολογίας ταυτόχρονης σακχαροποίησης και ζύμωσης (SSF). Οι ενζυμικές ενεργότητες που εφαρμόζονται στο στάδιο των ζυμώσεων είναι αυτές που παρουσιάζουν τα βέλτιστα αποτελέσματα κατά την υδρόλυση του υλικού. Η παραγωγικότητα της διεργασίας κυμάνθηκε από 0,08-0,26 g/L*h. Το εύρος τιμών παραγωγής αιθανόλης κατά την ζύμωση είναι 7,22±0,84 – 18,70±0,13g/L, ενώ τα ποσοστά % των μέγιστων θεωρητικών αποδόσεων είναι 68,87±1,09 και 288,30±4,43 βάσει της μέγιστης παραγόμενης αιθανόλης των μη δομικών σακχάρων, της κυτταρίνης και του αμύλου και βάσει της μέγιστης παραγόμενης αιθανόλης από τα μη δομικά σάκχαρα.The production of biofuels, particularly bioethanol, by biomass has been the center of scientific and industrial interest in the recent years. Specifically, in order for the production of second generation bioethanol to become sustainable, a considerable effort is placed into reducing the cost of raw materials. The management of food waste is a major concern in terms of protecting the environment and conserving the resources. The conversion of waste food into ethanol could potentially offer further cost advantages, due to the rising cost of waste disposal. The fact that food waste is rich in fermentable sugars, makes it suitable feedstock for bioethanol, an alternative and renewable transport fuel. The purpose of this thesis is to study the process of bioethanol production from food waste and optimize the respective production conditions. Initially, the raw material is pretreated hydrothermally at 100°C temperature for 1 hour with and without the presence of H2SO4 in solid content 1g/100g, with a concentration of solid of 30% w/v. The percentage of starch is degraded 47.13% after hydrothermal pretreatment with H2SO4, 1g/100g of substrate and 45.72% after hydrothermal pretreatment without acid. Enzymatic hydrolysis of the pretreated material is carried out using amylolytic (Liquozyme SC DS and Spirizyme Fuel) with activities 0,053 U / g of starch to alpha-amylase and 5.06, 10.2, 20.24 U / g starch to glucoamylase and cellulolytic (Celluclast 1,5 L / Novozyme 188 (5:1) and Cellic ® CTec2) commercially available enzymes with activities 1,3,5,7,9 FPU / g material. Starch hydrolysis is between 72.30% and 86.32%. The theoretical rate of hydrolysis of the cellulose, based in the produced glucose during the hydrolysis with mixture of Celluclast 1,5 L / Novozyme 188 (5:1) is between 75.28% and 97.63%, while regarding the hydrolysis by using the formula Cellic ® CTec2 the figure is 25.70% to 64.22%. Finally, it is studied the behavior of amylolytic and cellulolytic enzyme formulas and their synergistic effect against the production of ethanol by application of the technology of simultaneous saccharification and fermentation (SSF). The enzyme activities applied at the stage of fermentation are those that show the best results in the hydrolysis of the material. The process productivity ranged from 0,08-0,26 g / L * h. The range of ethanol production during fermentation is 7,22 ± 0,84 - 18,70 ± 0,13 g / L, while the percentage of the maximum theoretical yield is 68,87 ± 1,09 and 288,30 ± 4, 43 based on the maximum of the produced ethanol from nonstructural sugars, cellulose and starch and the maximum ethanol produced from the non-structural sugars.Γεωργία Ν. Παπαδοπούλο

Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation

Characterization of the c.793-1G > A splicing variant in CHEK2 gene as pathogenic: a case report

BackgroundCHEK2 is involved in the DNA damage repair response Fanconi anemia (FA)-BRCA pathway. An increased risk for breast and other cancers has been documented in individuals who carry a single pathogenic CHEK2 variant. As for other genes involved in cancer predisposition, different types of pathogenic variants have been observed, including single nucleotide variations, short insertions/deletions, large genomic rearrangements and splicing variants. Splicing variants occurring in the splicing acceptor or donor site result in alternative mature mRNA produced and can cause intron retention, exon skipping, or creation of alternative 3 and 5 splice site. Thus, the pathogenicity of this type of alterations should always be explored experimentally and their effect in the mRNA and consequently the protein produced, should be defined. The aim of this study was the delineation of the effect of a splicing variant in the CHEK2 gene.Case presentation A healthy 28-year-old woman with a family history of breast and ovarian cancer was referred for genetic testing. The variant c.793-1G>A (rs730881687) was identified by Next Generation Sequencing (NGS) using a solution-based capture method, targeting 33 cancer predisposition genes (SeqCap EZ Probe library, Roche NimbleGen). Experimental analysis in patient-derived leukocytes using RT-PCR of mRNA followed by cDNA sequencing revealed the deletion of one base from the alternative transcript created (r.793del). This resulted in a frameshift leading to premature termination codon within exon 7 (p.(Asp265Thrfs*10)).Conclusions This finding suggests that the CHEK2 splicing variant c.793-1G>A is a deleterious variant. Our case shows that RNA analysis is a valuable tool for uncharacterized splice site variants in individuals referred for testing and facilitates their personalized management

Report of a germline double heterozygote in MSH2 and PALB2

Abstract Background Carriers with pathogenic variants in MSH2 have increased risk to develop colorectal, endometrium, ovarian, and other types of cancer. The PALB2 is associated with breast, ovarian, pancreatic, and prostate cancer. We describe the case of a 42‐year‐old female diagnosed with endometrial cancer at the age of 42 years with a strong family history of colorectal cancer, which was referred to our private diagnostic laboratory for genetic testing. Methods In this study, we performed next‐generation sequencing (NGS) using an amplicon based 26 genes panel. The presence of multi‐exonic copy number variations (CNVs) was investigated by computational analysis and Multiplex Ligation‐dependent Probe Amplification (MLPA). Results A gross deletion of the genomic region encompassing exons 11–16 of the MSH2 and the loss‐of‐function variant c.757_758delCT, p.(Leu253Ilefs*3) in the PALB2 were identified in the proband. Conclusions Multigene analysis using NGS technology allows the identification of pathogenic variants in genes that would normally not be tested based on the patient diagnosis. In our case these results explained not only the personal and/or family history of cancer but also allowed the surveillance for prevention of other cancer types. Moreover, the detection of large genomic rearrangements should be routinely included in hereditary cancer testing

Synthesis and Characterization of Mesoporous Mg- and Sr-Doped Nanoparticles for Moxifloxacin Drug Delivery in Promising Tissue Engineering Applications

International audienceMesoporous silica-based nanoparticles (MSNs) are considered promising drug carriers because of their ordered pore structure, which permits high drug loading and release capacity. The dissolution of Si and Ca from MSNs can trigger osteogenic differentiation of stem cells towards extracellular matrix calcification, while Mg and Sr constitute key elements of bone biology and metabolism. The aim of this study was the synthesis and characterization of sol–gel-derived MSNs co-doped with Ca, Mg and Sr. Their physico-chemical properties were investigated by X-ray diffraction (XRD), scanning electron microscopy with energy dispersive X-ray analysis (SEM/EDX), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray fluorescence spectroscopy (XRF), Brunauer Emmett Teller and Brunauer Joyner Halenda (BET/BJH), dynamic light scattering (DLS) and ζ-potential measurements. Moxifloxacin loading and release profiles were assessed with high performance liquid chromatography (HPLC) cell viability on human periodontal ligament fibroblasts and their hemolytic activity in contact with human red blood cells (RBCs) at various concentrations were also investigated. Doped MSNs generally retained their textural characteristics, while different compositions affected particle size, hemolytic activity and moxifloxacin loading/release profiles. All co-doped MSNs revealed the formation of hydroxycarbonate apatite on their surface after immersion in simulated body fluid (SBF) and promoted mitochondrial activity and cell proliferation

One Step forward: Benthic Pelagic Coupling and Indicators for Environmental Status

<div><p>A large data set from the Eastern Mediterranean was analyzed to explore the relationship between seawater column variables and benthic community status. Our results showed a strong quantitative link between the seawater column variables (Chlorophyll <i>a</i> and Eutrophication Index) and various indicators describing benthic diversity and community composition. The percentage of benthic opportunistic species increased significantly in the stations with high trophic status of the seawater column and so did the strength of the coupling between values of seawater column and benthic indicators. The Eutrophication Index threshold level of 0.85, separating the “Bad and Poor” from “Moderate to High” conditions could serve as an acceptable critical value above which there is a readily observable change in benthic community composition.</p></div

Importance of multigene panel test in patients with consanguineous marriage and family history of breast cancer

Next-generation sequencing (NGS) technology is used to evaluate hereditary cancer risks of patients worldwide; however, information concerning the germline multigene mutational spectrum among patients with breast cancer (BC) with consanguineous marriage (CM) is limited. Therefore, this prospective study aimed to determine the molecular characteristics of patients with BC who were tested with multigene hereditary cancer predisposition NGS panel and to show the effect of CM on cancer-related genes. Patients with BC with or without CM and family history (FH) of BC treated in our breast center were selected according to The National Comprehensive Cancer Network (NCCN) criteria for hereditary BC. In these patients, the analysis of a panel of 33 genes involved in hereditary cancer predisposition was performed after genetic counseling by using NGS. The pathogenic variant (PV) and the variant of uncertain significance (VUS) were found to be 15.8 and 47.4%, respectively. PVs were identified in 10/33 genes in 34 patients; 38.2% in BRCA1/2 genes; 6, 24, and 14% in other high, moderate and low-risk genes, respectively. The CM rate was 17.7% among the 215 patients with BC. The PV rate was 13.2% in patients with CM and 16.4% in patients without CM (P=0.80). When PV and VUS were evaluated together, the PV+VUS ratio was significantly higher in patients with CM and FH of BC than patients without CM and FH of BC (88.2 vs. 63.3%, P=0.045). Analysis of multigene panel provided 9.76% additional PVs in moderate/low-risk genes. The PV rate was similar in patients with BC with or without CM. A high PV+VUS ratio in patients with CM and FH of BC suggests that genes whose importance are unknown are likely to be pathogenic genes later

Canonical Correspondence Analysis biplot with stations and environmental variables.

<p>Stations are labeled after the Ecological Status defined by means of the Benthic Quality Index—Family index: Symbols indicate Ecological Status: triangle, Bad; circle, Poor; square, Moderate; diamond, Good.</p

Number of sampling stations in each Ecological Status category according to four seawater column and benthic indices.

<p>Chl-<i>a</i>, chlorophyll <i>a</i>; EI, Eutrophic Index; BQI-Family, Benthic Quality Index—Family; BENTIX, Benthic Index.</p