Yield and content of biogenic elements in tomato fruit at different NaCl concentrations in nutrient solution

Kakvoću svježih plodova rajčice, osobito sadržaj minerala moguće je poboljšati povećanjem EC-vrijednosti, odnosno koncentracije NaCl-a u hranivoj otopini bez značajnijeg smanjenja prinosa. Istraživanje je provedeno tijekom 2004. i 2005. godine na sorti Belle, u hidroponima na pločama kamene vune s ciljem utvrđivanja utjecaja povećane koncentracije NaCl-a u hranivoj otopini na prinos rajčice i sadržaj biogenih elemenata u plodu. Pokus je postavljen po metodi slučajnog bloknog rasporeda u četiri ponavljanja, a uzorkovanje je provedeno u tri roka berbe. Hraniva otopina EC-vrijednosti 2 dS/m korištena je kao kontrola, a povećane ECvrijednosti su postignute dodavanjem NaCl-a u standardnu hranivu otopinu, 0,05 % pri 3 dS/m, 0,125 % pri 4,5 dS/m i 0,2 % pri 6 dS/m. Najveći prinos zabilježen je pri fertirigaciji standardnom hranivom otopinom i varirao je od 2,06 do 2,60 kg/biljci u 2004. i od 3,41 do 5,19 kg/biljci u 2005. Koncentracije 0,05 i 0,125 % NaCl-a rezultirale su statistički jednakim prinosom i sadržajem biogenih elemenata, a koncentracija 0,2 % NaCl-a signifikantno manjim vrijednostima navedenih parametara. Povećanjem koncentracije NaCl-a ostvareno je značajno povećanje sadržaja suhe tvari koje je proporcionalno povećanju NaCl-a u hranivoj otopini.Quality of fresh tomato fruits, especially mineral content can be improved by increasing the EC value, or the concentration of NaCl in the nutrient solution without a significant reduction in yield. The study was conducted during the years of 2004 and 2005 with Belle tomato variety in hydroponics rock wool slabs in order to determine the impact of increased concentrations of NaCl in the nutrient solution on tomato yield and content of biogenic elements in the fruit compared to the standard solution without added NaCl and the EC-value of 2 dS/m. The experiment was set in a randomized complete block design with four replications and sampling was conducted in three harvests. The nutrient solution with EC-value of 2 dS/m was used as a control and increased EC-values were achieved by adding NaCl in the standard nutrient solution, 0.05% at 3 dS/m, 0.125% at 4.5 dS/m and 0.2% at 6 dS/m. The highest yield was recorded with a standard nutrient solution treatment and ranged from 2.06 to 2.60 kg/plant in 2004 and from 3.41 to 5.19 kg/plant in 2004. In comparison to the standard solution without added NaCl, the concentration of 0.05 and 0.125% NaCl resulted in statistically equivalent yield and content of biogenic elements, and the concentration of 0.2% NaCl with reasonably smaller values of these parameters. Increasing NaCl recorded a significant increase in dry matter content, which is proportional to the increase of NaCl in the nutrient solution

Genetic lineage tracing reveals poor angiogenic potential of cardiac endothelial cells.

Abstract Aims Cardiac ischaemia does not elicit an efficient angiogenic response. Indeed, lack of surgical revascularization upon myocardial infarction results in cardiomyocyte death, scarring, and loss of contractile function. Clinical trials aimed at inducing therapeutic revascularization through the delivery of pro-angiogenic molecules after cardiac ischaemia have invariably failed, suggesting that endothelial cells in the heart cannot mount an efficient angiogenic response. To understand why the heart is a poorly angiogenic environment, here we compare the angiogenic response of the cardiac and skeletal muscle using a lineage tracing approach to genetically label sprouting endothelial cells. Methods and results We observed that overexpression of the vascular endothelial growth factor in the skeletal muscle potently stimulated angiogenesis, resulting in the formation of a massive number of new capillaries and arterioles. In contrast, response to the same dose of the same factor in the heart was blunted and consisted in a modest increase in the number of new arterioles. By using Apelin-CreER mice to genetically label sprouting endothelial cells we observed that different pro-angiogenic stimuli activated Apelin expression in both muscle types to a similar extent, however, only in the skeletal muscle, these cells were able to sprout, form elongated vascular tubes activating Notch signalling, and became incorporated into arteries. In the heart, Apelin-positive cells transiently persisted and failed to give rise to new vessels. When we implanted cancer cells in different organs, the abortive angiogenic response in the heart resulted in a reduced expansion of the tumour mass. Conclusion Our genetic lineage tracing indicates that cardiac endothelial cells activate Apelin expression in response to pro-angiogenic stimuli but, different from those of the skeletal muscle, fail to proliferate and form mature and structured vessels. The poor angiogenic potential of the heart is associated with reduced tumour angiogenesis and growth of cancer cells

Research of Fat Component Safety and Pre-Clinical Evaluation of Infant Adapted Dry Milk Mixtures Physiological Effect

The aim of the study deals with determination of fat component safety and quality key indicators of adapted infant dry milk formulas provided by various manufacturers. The most popular in Russia adapted infant dry milk formulas were selected as study objects. It was found that the qualitative composition of the fat component of dry milk mixtures corresponds to the information placed on the package. However none of the samples under study in terms of the average composition of the prevailing fatty acids fully corresponds to human breast milk. The regulation documents of the Customs Union (TR CU 021/2011, TR CU 024/2011, TR CU 033/2013) establish only the organoleptic evaluation of the adapted breast milk formulas quality indicators. Among the fat component safety indicators only the determination of the peroxide value characterizing the accumulation of primary fat oxidation products. It was also found that the peroxide values of the studied mixtures do not exceed the regulated values. Meanwhile the samples of infant milk food made from dry milk mixtures almost all have unsatisfactory organoleptic characteristics. Defects of taste and smell are associated with the accumulation in the original adapted milk mixtures of a significant amount of secondary products of fat oxidation, which in a biological experiment on animals lead to a decrease in the content of leukocytes and a change of its blood count

High-Throughput Screening to Enhance Permissiveness of Cardiac Endothelial Cells to Viral Vectors

Endothelial cells (ECs) are key players in the process of new blood vessel formation, also named angiogenesis, during both development and adulthood. Impairment of their function leads to many pathological conditions with either increased or inadequate vascularization. Thus, modulation of EC biology represents an attractive target in both basic and applied research. For instance, the major obstacle in inducing therapeutic angiogenesis is inefficient gene transfer in cardiac ECs. This poor permissiveness to genetic modification, attributed to their primary barrier function, is a limiting step in studies of pathological conditions with inadequate and increased vascularization. To study the role of a specific gene and its potential implications in angiogenesis, the most straightforward approach would be to either overexpress or silence it in ECs. We and others have tested multiple protocols for either transfection and transduction of primary ECs in vitro and in vivo, with minimal success. Vectors based on adeno-associated virus (AAV) and lentivirus (LV) stand as ideal tools for gene delivery. However, they do not work in cardiac ECs. To identify compounds able to increase transduction efficiency and identify the major determinants inhibiting genetic modification of cardiac ECs, we performed high-throughput screening (HTS) using a library of FDA-approved drugs in combination with both AAV and LV vectors. Cardiac ECs were isolated from C57BL/6 mice and plated in 384 wells plates. After 48 hours, the FDA drug library (> 1500 compounds) was added together with AAV6dsRed and LVGFP. At 72 hours after transduction, cells were fixed with 4% PFA and stained for the EC marker ERG. Expression of dsRed, GFP, and ERG was assessed by automated, high-content microscopy. After two rounds of HTS at different drug concentrations, we obtained a list of candidate drugs able that increased adult cardiac EC transduction by either AAV6 or LV over 2-fold. Among these, our best hit increasing cardiac EC transduction by AAV6 was vatalanib, a tyrosine kinase inhibitor acting on multiple VEGFRs. Vatalanib significantly enhanced permissiveness to AAV6 and also changed the shape of cardiac ECs, which became elongated and similar to mesenchymal cells. This suggested ongoing endothelial to mesenchymal transition (EndMT), verified by decreased CD31 and increased a-SMA expression levels. To validate our hypothesis, we induced EndMT by different methods, such as siRNA-mediated silencing of 2 EC-specific genes (CD31, ERG) in cardiac ECs and treatment of cardiac ECs with transforming growth factor-β2 (TGF-ß2). They all promoted loss of endothelial and acquisition of mesenchymal markers. Interestingly, AAV6 invariably transduced cells with the lowest expression of EC markers. Since cardiac ECs undergo transient EndMT early after myocardial infarction (MI), we injected AAV9-Cre into the heart of the mT/mG Cre-reporter mouse. After three days post-MI, cardiac ECs exhibiting EndMT features within the fibrotic zone were selectively transduced by AAV. Collectively, our results point to transient EndMT as a powerful mechanism to improve the efficiency of gene transfer in cardiac ECs, paving the way to innovative strategies for the induction of therapeutic angiogenesis in the heart

Razumijevanje slabe sposobnosti angiogeneze odraslog srca

Angiogenesis is the process of new blood vessel formation, crucial for the development of an embryo as well as for organ growth and wound healing later in the adulthood. A key factor responsible for the stimulation of angiogenesis is VEGF165, the most potent splicing isoform of VEGF-A. Based on recent results obtained in the host laboratory, the driving hypothesis standing beyond this work was to investigate why endothelial cells in the skeletal muscle and in the neonatal heart respond to AAV-VEGF165 inducing angiogenesis, whereas in the adult heart they do not. For this purpose, we investigated the responsiveness of adult cardiac and neonatal endothelial cells (ECs), as well as of muscle ECs to the proangiogenic factor VEGF in vitro. Based on the observation that cardiac ECs lose their proliferative capacity at day 7 after birth, we performed a transcriptome analysis of the three EC populations and identified a few differentially expressed genes (DEGs). Particularly, we focused on Dhcr24, the most interesting gene among DEG between pre-natal and post-natal ECs. We developed a genome editing strategy to investigate its functional role in angiogenesis. In the first set of experiments, the proliferation of adult and neonatal cardiac ECs and muscle ECs was assessed after VEGF administration at different time points, followed by immunofluorescence staining and image acquisition. Interestingly and different from their in vivo behaviour, adult cardiac ECs proliferated in response to VEGF at both time points, suggesting that some inhibitory factor might block their response in the adult heart in vivo. In the second set of experiments, the CRISPR/Cas9 technology was used to silence Dhcr24 in SVEC and LG cell lines. Successful knockout was confirmed by the downregulation of Dhcr24 protein expression in Western blot. Development of an efficient genome editing strategy could offer an opportunity to genetically modify Dhcr24 in quiescent ECs and test its ability to restore the pro-angiogenic phenotype.Angiogeneza je proces nastajanja novih krvnih žila, ključan tijekom razvoja embrija, te rasta organa i zacjeljivanja rana u odrasloj dobi. Ključni čimbenik koji je odgovoran za stimulaciju angiogeneze je VEGF165, najpotentnija izoforma alternativnog prekrajanja VEGF-A. Na temelju nedavno dobivenih rezultata u našem laboratoriju, glavna hipoteza ovog rada bila je istražiti zašto endotelne stanice (ES) u skeletnim mišićima i u neonatalnom srcu reagiraju na AAV-VEGF165 induciranu angiogenezu, dok one u odraslom srcu ne. U tu smo svrhu istražili odgovor odraslih i neonatalnih srčanih ES, kao i mišićnih ES mišića na proangiogeni čimbenik VEGF in vitro. Na temelju opažanja da srčane ES gube sposobnost proliferacije 7. dan nakon rođenja, izvršili smo transkripcijsku analizu tri populacije ES i identificirali nekoliko diferencijalno eksprimiranih gena (DEG). Posebno smo se usredotočili na Dhcr24, najzanimljiviji gen između DEG između prenatalnih i postnatalnih ES. Razvili smo strategiju uređivanja genoma kako bismo istražili njegovu funkcionalnu ulogu u angiogenezi. U prvoj skupini eksperimenata, proliferacija odraslih i neonatalnih srčanih ES i mišićnih ES određena je nakon administracije VEGF-a u različitim vremenskim točkama, nakon čega je slijedila imunofluorescencija i mikroskopija. Zanimljivo, za razliku od ponašanja in vivo, odrasle srčane ES su proliferirale kao odgovor na VEGF u obje vremenske točke, sugerirajući prisustvo nekog inhibitornog čimbenika koji može blokirati odgovor na VEGF u odraslom srcu in vivo. U drugoj skupini eksperimenata, CRISPR/Cas9 tehnologija korištena je za utišavanje Dhcr24 u SVEC i LG staničnim linijama. Uspješan knockout potvrđen je smanjenjem ekspresije Dhcr24 proteina u Western blotu. Razvijanje učinkovite strategije za uređivanje genoma moglo bi pružiti priliku za genetsko modificiranje Dhcr24 u ES u stanju mirovanja te ispitati ima li sposobnost vraćanja proangiogenog fenotipa

Razumijevanje slabe sposobnosti angiogeneze odraslog srca

Angiogenesis is the process of new blood vessel formation, crucial for the development of an embryo as well as for organ growth and wound healing later in the adulthood. A key factor responsible for the stimulation of angiogenesis is VEGF165, the most potent splicing isoform of VEGF-A. Based on recent results obtained in the host laboratory, the driving hypothesis standing beyond this work was to investigate why endothelial cells in the skeletal muscle and in the neonatal heart respond to AAV-VEGF165 inducing angiogenesis, whereas in the adult heart they do not. For this purpose, we investigated the responsiveness of adult cardiac and neonatal endothelial cells (ECs), as well as of muscle ECs to the proangiogenic factor VEGF in vitro. Based on the observation that cardiac ECs lose their proliferative capacity at day 7 after birth, we performed a transcriptome analysis of the three EC populations and identified a few differentially expressed genes (DEGs). Particularly, we focused on Dhcr24, the most interesting gene among DEG between pre-natal and post-natal ECs. We developed a genome editing strategy to investigate its functional role in angiogenesis. In the first set of experiments, the proliferation of adult and neonatal cardiac ECs and muscle ECs was assessed after VEGF administration at different time points, followed by immunofluorescence staining and image acquisition. Interestingly and different from their in vivo behaviour, adult cardiac ECs proliferated in response to VEGF at both time points, suggesting that some inhibitory factor might block their response in the adult heart in vivo. In the second set of experiments, the CRISPR/Cas9 technology was used to silence Dhcr24 in SVEC and LG cell lines. Successful knockout was confirmed by the downregulation of Dhcr24 protein expression in Western blot. Development of an efficient genome editing strategy could offer an opportunity to genetically modify Dhcr24 in quiescent ECs and test its ability to restore the pro-angiogenic phenotype.Angiogeneza je proces nastajanja novih krvnih žila, ključan tijekom razvoja embrija, te rasta organa i zacjeljivanja rana u odrasloj dobi. Ključni čimbenik koji je odgovoran za stimulaciju angiogeneze je VEGF165, najpotentnija izoforma alternativnog prekrajanja VEGF-A. Na temelju nedavno dobivenih rezultata u našem laboratoriju, glavna hipoteza ovog rada bila je istražiti zašto endotelne stanice (ES) u skeletnim mišićima i u neonatalnom srcu reagiraju na AAV-VEGF165 induciranu angiogenezu, dok one u odraslom srcu ne. U tu smo svrhu istražili odgovor odraslih i neonatalnih srčanih ES, kao i mišićnih ES mišića na proangiogeni čimbenik VEGF in vitro. Na temelju opažanja da srčane ES gube sposobnost proliferacije 7. dan nakon rođenja, izvršili smo transkripcijsku analizu tri populacije ES i identificirali nekoliko diferencijalno eksprimiranih gena (DEG). Posebno smo se usredotočili na Dhcr24, najzanimljiviji gen između DEG između prenatalnih i postnatalnih ES. Razvili smo strategiju uređivanja genoma kako bismo istražili njegovu funkcionalnu ulogu u angiogenezi. U prvoj skupini eksperimenata, proliferacija odraslih i neonatalnih srčanih ES i mišićnih ES određena je nakon administracije VEGF-a u različitim vremenskim točkama, nakon čega je slijedila imunofluorescencija i mikroskopija. Zanimljivo, za razliku od ponašanja in vivo, odrasle srčane ES su proliferirale kao odgovor na VEGF u obje vremenske točke, sugerirajući prisustvo nekog inhibitornog čimbenika koji može blokirati odgovor na VEGF u odraslom srcu in vivo. U drugoj skupini eksperimenata, CRISPR/Cas9 tehnologija korištena je za utišavanje Dhcr24 u SVEC i LG staničnim linijama. Uspješan knockout potvrđen je smanjenjem ekspresije Dhcr24 proteina u Western blotu. Razvijanje učinkovite strategije za uređivanje genoma moglo bi pružiti priliku za genetsko modificiranje Dhcr24 u ES u stanju mirovanja te ispitati ima li sposobnost vraćanja proangiogenog fenotipa

Razumijevanje slabe sposobnosti angiogeneze odraslog srca

Angiogenesis is the process of new blood vessel formation, crucial for the development of an embryo as well as for organ growth and wound healing later in the adulthood. A key factor responsible for the stimulation of angiogenesis is VEGF165, the most potent splicing isoform of VEGF-A. Based on recent results obtained in the host laboratory, the driving hypothesis standing beyond this work was to investigate why endothelial cells in the skeletal muscle and in the neonatal heart respond to AAV-VEGF165 inducing angiogenesis, whereas in the adult heart they do not. For this purpose, we investigated the responsiveness of adult cardiac and neonatal endothelial cells (ECs), as well as of muscle ECs to the proangiogenic factor VEGF in vitro. Based on the observation that cardiac ECs lose their proliferative capacity at day 7 after birth, we performed a transcriptome analysis of the three EC populations and identified a few differentially expressed genes (DEGs). Particularly, we focused on Dhcr24, the most interesting gene among DEG between pre-natal and post-natal ECs. We developed a genome editing strategy to investigate its functional role in angiogenesis. In the first set of experiments, the proliferation of adult and neonatal cardiac ECs and muscle ECs was assessed after VEGF administration at different time points, followed by immunofluorescence staining and image acquisition. Interestingly and different from their in vivo behaviour, adult cardiac ECs proliferated in response to VEGF at both time points, suggesting that some inhibitory factor might block their response in the adult heart in vivo. In the second set of experiments, the CRISPR/Cas9 technology was used to silence Dhcr24 in SVEC and LG cell lines. Successful knockout was confirmed by the downregulation of Dhcr24 protein expression in Western blot. Development of an efficient genome editing strategy could offer an opportunity to genetically modify Dhcr24 in quiescent ECs and test its ability to restore the pro-angiogenic phenotype.Angiogeneza je proces nastajanja novih krvnih žila, ključan tijekom razvoja embrija, te rasta organa i zacjeljivanja rana u odrasloj dobi. Ključni čimbenik koji je odgovoran za stimulaciju angiogeneze je VEGF165, najpotentnija izoforma alternativnog prekrajanja VEGF-A. Na temelju nedavno dobivenih rezultata u našem laboratoriju, glavna hipoteza ovog rada bila je istražiti zašto endotelne stanice (ES) u skeletnim mišićima i u neonatalnom srcu reagiraju na AAV-VEGF165 induciranu angiogenezu, dok one u odraslom srcu ne. U tu smo svrhu istražili odgovor odraslih i neonatalnih srčanih ES, kao i mišićnih ES mišića na proangiogeni čimbenik VEGF in vitro. Na temelju opažanja da srčane ES gube sposobnost proliferacije 7. dan nakon rođenja, izvršili smo transkripcijsku analizu tri populacije ES i identificirali nekoliko diferencijalno eksprimiranih gena (DEG). Posebno smo se usredotočili na Dhcr24, najzanimljiviji gen između DEG između prenatalnih i postnatalnih ES. Razvili smo strategiju uređivanja genoma kako bismo istražili njegovu funkcionalnu ulogu u angiogenezi. U prvoj skupini eksperimenata, proliferacija odraslih i neonatalnih srčanih ES i mišićnih ES određena je nakon administracije VEGF-a u različitim vremenskim točkama, nakon čega je slijedila imunofluorescencija i mikroskopija. Zanimljivo, za razliku od ponašanja in vivo, odrasle srčane ES su proliferirale kao odgovor na VEGF u obje vremenske točke, sugerirajući prisustvo nekog inhibitornog čimbenika koji može blokirati odgovor na VEGF u odraslom srcu in vivo. U drugoj skupini eksperimenata, CRISPR/Cas9 tehnologija korištena je za utišavanje Dhcr24 u SVEC i LG staničnim linijama. Uspješan knockout potvrđen je smanjenjem ekspresije Dhcr24 proteina u Western blotu. Razvijanje učinkovite strategije za uređivanje genoma moglo bi pružiti priliku za genetsko modificiranje Dhcr24 u ES u stanju mirovanja te ispitati ima li sposobnost vraćanja proangiogenog fenotipa

The safety study of the fat component in adapted infant formula

The growth and harmonious development of an infant depends on nutrition provided by natural or artificial feeding. Accordingly the adaptation of the composition of adapted infant formula to the composition of human milk is carried out with all major nutrients. The fat component is one of the key components of an infant’s diet and its’ important aspects are biological effectiveness and safety which changes during processing, storage, transportation and subsequent storage after opening the package. Data on pathological changes in the body with consumption of oxidized fats are known. The aim of the paper is to study some indicators of safety and quality of the fat component of adapted infant formula provided by various manufacturers. As the objects of study we selected one of the most popular adapted infant formula in Russia (in various price categories) under the code names: “IS” (Denmark), “IM”, “IN”, “IL” (Russian Federation), “IX “(Germany). It was found that the quality composition of the fat component of dry infant formula corresponds to the one indicated on the package. However, no one of the test samples according to the averaged composition of the prevailing fatty acids, is fully identical to human milk. The normative documentation of the Customs Union (TR TS 021/2011, TP TS 024/2011, TP TS 033/2013) only regulates organoleptic analysis of the quality indicators of adapted infant formula and from the safety parameters, only finding the peroxide value, which characterizes the accumulation of primary fat oxidation products. It was found that the peroxide numbers of the studied infant formula do not exceed the regulated values. At the same time almost all milk nutrition made from dry infant formula has unsatisfactory organoleptic characteristics. Defects of taste and aroma can be associated with the accumulation of a significant amount of secondary products of fat oxidation, aldehydes particularly in the adapted infant formula. These conclusions are confirmed by the results of determination of the anisidine number in the adapted infant formula, as well as by the accumulation of 0.3-1.0% of highly polar compounds insoluble in petroleum ether (CIPE) and 25-3 mmol/kg of epoxides which lead to decrease of white blood cells and a change in blood formula at a biological animal test. The data obtained indicate the need for assessment of the technology for the production, packaging and storage of adapted infant formula as well as the feasibility of amending the regulatory documents of the Russian Federation and in the future, the regulatory documents of the Customs Union with the aim of further monitoring the safety of the fat component of adapted infant formula

Changes in event-related synchronization/desynchronization of brain electric activity in cardiosurgical patients with postoperative cognitive dysfunction

Aim of the study was to analyze the event-related synchronization/desynchronization of brain electrical activity during visual selection task in patients underwent on-pump coronary artery bypass grafting (CABG) with and without postoperative cognitive dysfunction (POCD). Material and methods. The study included 32 men who underwent on-pump CABG, mean age 57,2 ± 6,08 years. All patients carried out extended neuropsychological testing, a multi-channel computer electroencephalography (EEG) 3-5 days before CABG and on the 7-10th day after the surgery. The POCD was determined according to the criterion: 20 % decrease in the cognitive indicator compared to that at baseline on 20 % of the tests included in the neuropsychological battery. Monopolar EEGs were recorded in 62 sites of 10-20 system with NEUVO encephalograph (Compumedics, USA) during cognitive task performing in patients with and without POCD. Statistical processing was performed using the STATISTICA 10.0. Results. It was found that the POCD patients had less pronounced theta desynchronization in the left frontal-central regions during the stage of 200-400 ms at the 7-10 days after CABG in comparison to patients without cognitive decline. Moreover, in the left parietal leads POCD patients had decreased theta desynchronization during the stage of 200-400 ms even before the surgery. At the 7-10 days after CABG, only the patients without POCD had a decrease of event-related theta activity in the left parietal leads compared with baseline. During the stage of 600-800 ms, the POCD patients had a lower degree of theta-desynchronization of both frontal-central and parietal regions of right hemisphere compared to patients without cognitive decline. Conclusion. The cognitive decline in patients after CABG determined according to neuropsychological testing is accompanied by pathological changes in the event-related theta activity. An analysis of event-related synchronization/desynchronization can be used both as predictor of postoperative cognitive impairment and as objective marker of POCD