Ecological production types of the Radava forests

Основни задатак истраживања је да се на основу показатеља о карактеристикама станишта и састојина на територији планине Радава дефинишу производни типови шума у чистим буковим шумама, затим у мјешовитим шумама букве и јеле, шумама букве, јеле и смрче, у шумама јеле и смрче, као и у мјешовитим шумама букве, јеле, смрче и Панчићеве оморике. Дефинисањем типова шума створиће се реални услови за утврђивање циљева газдовања шумама у овим екосистемима. Истраживања полазе од основне дефиниције типа шуме који представља дио шуме са подједнаким еколошко - биолошким и развојно - производним карактеристикама. Критеријуми за избор састојина у којима су вршена истраживања су једноличност станишних и хомогеност састојинских услова, као што су надморска висина, нагиб, склоп, експозиција и др., а циљ је да се обухвате разноликости састојина и да услови у оквиру огледних површина буду једнолични. Укупно је постављено 24 огледна поља просјечне величине око 0,5 ha. У шумама букве постављено је 8 огледних површина, шумама букве и јеле 4, шумама букве, јеле и смрче 4, шумама јеле и смрче 4 и у шумама букве, јеле, смрче и Панчићеве оморике 4 огледне површине. Обрада и анализа података је извршена примјеном уобичајених дендрометријских и статистичких метода. На основу резултата еколошких истраживања дефинисано је 7 основних типова шума. Производним диференцирањем основних типова шума је издвојено 6 производних типова шума: производни тип шуме 1 – шума букве на станишту букве (Fagetum montanum illyricum Fuk. et Stef. 1958) на серији земљишта на кречњаку, производни тип шуме 2 – шума букве и јеле на станишту букве и јеле (Abieti - Fagetum illyricum Fuk. et Stef. 1958) на серији земљишта на кречњаку, претежно дубоких, производни тип шуме 3 – шума букве, јеле и смрче на станишту букве, јеле и смрче (Piceo – Abieti - Fagetum Stef. et Beus 1983) на серији земљишта на кречњаку, претежно дубоких земљишта, производни тип шуме 4 – шума букве, јеле, смрче и Панчићеве оморике (Piceo-Abieti-Fagetum omorikae) на серији земљишта на кречњаку, производни тип шуме 5 – шума јеле и смрче на станишту јеле и смрче (Abieti-Picetum Mat. 1978) на дистричном смеђем земљишту на пјесковитом глинцу, производни тип шуме 6 – шума јеле и смрче на станишту јеле и смрче (Abieti-Picetum Mat. 1978) на серији земљишта на кречњаку, претежно дубоких.The basic task of the research is to define the production types of forests in pure beech forests, then in mixed forests of beech and fir, beech, fir and spruce forests, in fir and spruce forests as well as in the mixed forests of beech, fir, spruce and Serbian spruce (Pancic spruce) in accordance with indicators on the characteristics of the habitat and stands in the territory of the Radava mountain. Defining the types of forest will create real conditions for determining forest management objectives in these ecosystems. The research is based on the basic definition of the forest type that represents a part of the forest with equal ecological-biological and developmental-production characteristics. The criteria for selecting the stands in which the research was carried out are the uniformity of the habitat and homogeneity of the stand conditions, such as altitude, inclination, composition, exposition, etc., with the aim to include the variety of stands and that the conditions within the experimental surfaces are uniform. A total of 24 experimental fields of an average size of about 0.5 ha were placed. There were 8 experimental surfaces in the beech forests, 4 in beech and fir forests, 4 in beech, fir and spruce forests, 4 in fir and spruce forests and 4 in beech, fir, spruce and Serbian spruce forests. Data processing and analysis was carried out using the usual dendrometric and statistical methods. Based on the results of ecological research, 7 basic types of forest are defined. The production differentiation of the basic types of forests separated 6 production types of forests: production forest type 1 - beech forest on beech habitat on limestone soil series, production forest type 2 - beech and fir forest on beech and fir habitat on limestone soil series, predominantly deep soils, production forest type 3 - beech, fir and spruce forest on beech, fir and spruce habitat on limestone soil series, predominantly deep soils, production forest type 4 - beech, fir, spruce and Serbian spruce forest on a limestone soil series, production forest type 5 - fir and spruce forest on fir and spruce habitat on dystric brown soil on sandy clay, production forest type 6 - fir and spruce forest on fir and spruce habitat on limestone soil series, predominantly deep soil

Molecular basis of mitochondrial structural remodeling induced by calcium and insulin in rat brown adipocytes

Osnovna uloga mrkog masnog tkiva je održanje tjelesne temperature i održanje energetske homeostaze organizma. Izlaganje hladnoći, a i povećan unos hrane, dovodi do stimulacije, proliferacije i diferencijacije mrkih adipocita, što je praćeno remodeliranjem mitohondrija, termogenih organela mrkih adipocita. U okviru strukturnog remodeliranja mitohondrije pokazuju širok dijapazon morfoloških promjena (broj mitohondrija u ćeliji, srednji dijametar, fuzija i fisija mitohondrija, volumenska gustina kristi) što je povezano sa prisustvom pojedinih kompleksa elektron transportnog lanca, ATP sintaze, familije UCP proteina, posebno UCP1, a i drugih proteina unutrašnje membrane mitohondrija. Kalcijum igra veoma važnu ulogu u termogenezi, njegova koncentracija u citoplazmi i organelama mrkih adipocita je hormonski regulisana, prije svega noradrenalinom. Mitohondrije, s jedne strane, imaju važnu ulogu u oblikovanju signalnih puteva kalcijuma, a kalcijum, s druge strane, u fiziološkim koncentracijama stimuliše mitohondrijalni metabolizam i povećava produkciju energije, dok u visokim koncentracijama indukuje apoptozu. Insulin je jedan od glavnih modulatora termogene funkcije mrkih adipocita, gdje stimuliše sintezu i deponovanje lipida u mrkim adipocitima, proliferaciju mrkih adipocita, angiogenezu i vazodilataciju. Pokazano je da insulin utiče na aktivnost elektron transportnog lanca, a i da mitohondrije pojačavaju insulinsku osjetljivost putem redoks regulacije proteina tirozin kinaze i insulinskog receptora. Cilj ove doktorske disertacije je rasvjetljavanje kako kalcijum, s jedne strane, a insulin, s druge strane, utiču na molekulske mehanizme koje su uključene u strukturno remodeliranje mitohondrija mrkih adipocita pacova. U eksperimentu su korišćeni Wistar pacovi, starosti dva mjeseca. U eksperimentu sa Ca-SANDOZ, životinje su podijeljene u dve grupe, jedna je pila Ca-SANDOZ rastvoren u vodi, a druga, kontrolna, česmensku vodu, tri dana. U eksperimentu sa insulinom, pacovi su podijeljeni u šest grupa, četiri grupe su primale nisku (0.4 IU) ili visoku (4 IU) dozu insulina i to jedan (akutno) ili tri dana (hronično). Ostale dvije grupe, kontrolne, su primale fiziološki rastvor akutno ili hronično. Trećeg dana tretmana Ca-SANDOZ, odnosno tri sata poslije posljednjeg tretmana insulinom, životinje su žrtvovane dekapitacijom, a interskapularni depo BAT je izolovan i pripremljen za biohemijske, mikroskopske i molekularno-biološke analize, i za izolaciju mitohondrijalnih frakcija. BAT i mitohondrijama bogata frakcija su analizirane Western Blot analizom, imunohistohemijskom, imunofluorescentnom i imunogold analizama, ultrastukturnim i stereološkim analizama, a analizirana je i aktivnost ATP sintaze...Essential role of brown adipose tissue is maintaining body temperature and energy homeostasis of organism. Exposure to cold and increased diet intake lead to stimulation, proliferation and differentiation of brown adipocytes, what is followed by remodeling of mitochondria, thermogenic organelles of brown adipocytes. In the context of structural remodeling, mitochondria show wide repertoire of morphological changes (e.g. mitochondria number in cell, mean diameter, fusion and fission of mitochondria, volume density of mitochondrial cristae) what is connected with abundance of components of electron transport chain (ETC), ATP synthase, UCP proteins - especially UCP1 protein, and other proteins of inner mitochondrial membrane. Calcium plays important role in thermogenesis, and its concentration in cytosol and organelles of brown adipocytes is hormone regulated, particularly by noradrenaline. Mitochondria has essential role in shaping calcium signaling pathways, and calcium, on other hand, stimulates mitochondrial metabolism and increases energy production, but in high concentrations induces apoptosis. Insulin is one of major modulators of thermogenic function of brown adipocytes, and stimulates synthesis and storage of lipids in brown adipocytes, proliferation of brown adipocytes, angiogenesis and vasodilatation. It is showed that insulin has effect on electron transport chain activity, but also mitochondria enhances insulin sensitivity via redox regulation of tyrosine kinases and insulin receptor. The aim of this doctoral dissertation is elucidation how calcium and insulin effect on molecular mechanisms involved in structural remodeling of mitochondria of rat brown adipocytes. Two-month-old male Wistar rats were used. In Ca-SANDOZ study, they were divided into two groups - Ca-SANDOZ drinking or tap water drinking for three days. In insulin experiment, rats were divided into six groups, where four groups treated with low (0.4 IU) or high (4 IU) dose of insulin, for one (acutely) or three (chronically) days. Another two groups served as control, treated with 0.9% saline solution for one or three days. On third day of Ca-SANDOZ treatment, or three hours after last administration of insulin, they were sacrificed by decapitation, and interscapular portion of BAT was isolated and prepared for biochemical, microscopic and molecular biology analyses, also for isolation of mitochondria-enriched BAT fraction. BAT and mitochondria-enriched fraction were analyzed by Western blot analysis, imunohistochemical, immunofluorescent, immunogold, ultrastructural and stereological analyses. Also, ATP synthase activity was measured..

The effects of biocides on the growth of aerophytic green algae (Chlorella sp.) isolated from a cave environment

Lampenflora communities of bacteria, cyanobacteria, algae, mosses and lichens colonize illuminated show-caves as a consequence of disturbances to the caves’ ecological equilibrium. These communities have unesthetic impacts and can cause the biodeterioration of limestone. A 15% hydrogen peroxide solution was proposed for use as an ecological agent for the safe removal of these microorganisms. This study tested the effects of three different biocides (hydrogen peroxide, a commercial solution containing salicylic acid and a product containing chlorine dioxide as the active component), on the growth of green algae (Chlorella sp.), which are frequently encountered in lampenflora communities. Chlorella sp. was treated with the biocides under laboratory condition, and chlorophyll a (Chl a) concentrations were measured 1 h, 3 days, 7 days and 10 days after treatment. The change in Chl a concentration was compared to the untreated control group at each time point. All three biocides prevented the growth of green algae and the product containing chlorine dioxide appeared to be the most effective growth inhibitor. Hydrogen peroxide is known to suppress lampenflora growth in caves, and further studies on other biocides remain necessary to identify a solution that is both ecologically safe and economically feasible

Promotion of Early Gut Colonization by Probiotic Intervention on Microbiota Diversity in Pregnant Sows

The aim of this work was to design a novel mixed probiotic culture for piglets and to evaluate its beneficial effect on the piglets' gut health. The possible mechanisms of probiotic activity, such as adhesion, competitive pathogen exclusion and influence on gut microbiota diversity were determined. Mixed probiotic starter culture is composed of three thermophilic lactic acid bacteria (LAB) strains: Lactobacillus helveticus BGRA43, Lactobacillus fermentum BGHI14 and Streptococcus thermophilus BGVLJ1-44. The strains BGVLJ1-44 and BGRA43 showed good technological properties (fast milk curdling, strong proteolytic activity). In addition, the strain BGVLJ1-44 produces exopolysaccharide (EPS), BGHI14 is heterofermentative LAB strain with significant immunomodulatory effect, while the strain BGRA43 showed strong antimicrobial activity against different pathogens and exhibited significantly higher level of adhesion to Caco-2 cells comparing to other two strains. Both lactobacilli strains BGRA43 and BGHI14 (p lt 0.05), as well as probiotic combination (p lt 0.01) significantly reduced the adhesion of Escherichia coli ATCC25922 to Caco-2 cells, while the strains BGVLJ1-44 (p lt 0.01) and BGRA43 (p lt 0.05) significantly reduced adhesion of Salmonella 654/7E (veterinary isolate). The results of farm trial revealed that treatment of sows with new fermented dairy probiotic influenced the piglets' gut colonization with beneficial bacteria and reduced the number of enterobacteriaceae in litters from some treated sows (no significant due to high variability among animals). Finally, this is the first study reporting that the treatment of sows with probiotic combination resulted in the improved microbiota diversity in neonatal piglets

Ecological production types of the Radava forests

Основни задатак истраживања је да се на основу показатеља о карактеристикама станишта и састојина на територији планине Радава дефинишу производни типови шума у чистим буковим шумама, затим у мјешовитим шумама букве и јеле, шумама букве, јеле и смрче, у шумама јеле и смрче, као и у мјешовитим шумама букве, јеле, смрче и Панчићеве оморике. Дефинисањем типова шума створиће се реални услови за утврђивање циљева газдовања шумама у овим екосистемима. Истраживања полазе од основне дефиниције типа шуме који представља дио шуме са подједнаким еколошко - биолошким и развојно - производним карактеристикама. Критеријуми за избор састојина у којима су вршена истраживања су једноличност станишних и хомогеност састојинских услова, као што су надморска висина, нагиб, склоп, експозиција и др., а циљ је да се обухвате разноликости састојина и да услови у оквиру огледних површина буду једнолични. Укупно је постављено 24 огледна поља просјечне величине око 0,5 ha. У шумама букве постављено је 8 огледних површина, шумама букве и јеле 4, шумама букве, јеле и смрче 4, шумама јеле и смрче 4 и у шумама букве, јеле, смрче и Панчићеве оморике 4 огледне површине. Обрада и анализа података је извршена примјеном уобичајених дендрометријских и статистичких метода. На основу резултата еколошких истраживања дефинисано је 7 основних типова шума. Производним диференцирањем основних типова шума је издвојено 6 производних типова шума: производни тип шуме 1 – шума букве на станишту букве (Fagetum montanum illyricum Fuk. et Stef. 1958) на серији земљишта на кречњаку, производни тип шуме 2 – шума букве и јеле на станишту букве и јеле (Abieti - Fagetum illyricum Fuk. et Stef. 1958) на серији земљишта на кречњаку, претежно дубоких, производни тип шуме 3 – шума букве, јеле и смрче на станишту букве, јеле и смрче (Piceo – Abieti - Fagetum Stef. et Beus 1983) на серији земљишта на кречњаку, претежно дубоких земљишта, производни тип шуме 4 – шума букве, јеле, смрче и Панчићеве оморике (Piceo-Abieti-Fagetum omorikae) на серији земљишта на кречњаку, производни тип шуме 5 – шума јеле и смрче на станишту јеле и смрче (Abieti-Picetum Mat. 1978) на дистричном смеђем земљишту на пјесковитом глинцу, производни тип шуме 6 – шума јеле и смрче на станишту јеле и смрче (Abieti-Picetum Mat. 1978) на серији земљишта на кречњаку, претежно дубоких.The basic task of the research is to define the production types of forests in pure beech forests, then in mixed forests of beech and fir, beech, fir and spruce forests, in fir and spruce forests as well as in the mixed forests of beech, fir, spruce and Serbian spruce (Pancic spruce) in accordance with indicators on the characteristics of the habitat and stands in the territory of the Radava mountain. Defining the types of forest will create real conditions for determining forest management objectives in these ecosystems. The research is based on the basic definition of the forest type that represents a part of the forest with equal ecological-biological and developmental-production characteristics. The criteria for selecting the stands in which the research was carried out are the uniformity of the habitat and homogeneity of the stand conditions, such as altitude, inclination, composition, exposition, etc., with the aim to include the variety of stands and that the conditions within the experimental surfaces are uniform. A total of 24 experimental fields of an average size of about 0.5 ha were placed. There were 8 experimental surfaces in the beech forests, 4 in beech and fir forests, 4 in beech, fir and spruce forests, 4 in fir and spruce forests and 4 in beech, fir, spruce and Serbian spruce forests. Data processing and analysis was carried out using the usual dendrometric and statistical methods. Based on the results of ecological research, 7 basic types of forest are defined. The production differentiation of the basic types of forests separated 6 production types of forests: production forest type 1 - beech forest on beech habitat on limestone soil series, production forest type 2 - beech and fir forest on beech and fir habitat on limestone soil series, predominantly deep soils, production forest type 3 - beech, fir and spruce forest on beech, fir and spruce habitat on limestone soil series, predominantly deep soils, production forest type 4 - beech, fir, spruce and Serbian spruce forest on a limestone soil series, production forest type 5 - fir and spruce forest on fir and spruce habitat on dystric brown soil on sandy clay, production forest type 6 - fir and spruce forest on fir and spruce habitat on limestone soil series, predominantly deep soil

Molecular basis of mitochondrial structural remodeling induced by calcium and insulin in rat brown adipocytes

Osnovna uloga mrkog masnog tkiva je održanje tjelesne temperature i održanje energetske homeostaze organizma. Izlaganje hladnoći, a i povećan unos hrane, dovodi do stimulacije, proliferacije i diferencijacije mrkih adipocita, što je praćeno remodeliranjem mitohondrija, termogenih organela mrkih adipocita. U okviru strukturnog remodeliranja mitohondrije pokazuju širok dijapazon morfoloških promjena (broj mitohondrija u ćeliji, srednji dijametar, fuzija i fisija mitohondrija, volumenska gustina kristi) što je povezano sa prisustvom pojedinih kompleksa elektron transportnog lanca, ATP sintaze, familije UCP proteina, posebno UCP1, a i drugih proteina unutrašnje membrane mitohondrija. Kalcijum igra veoma važnu ulogu u termogenezi, njegova koncentracija u citoplazmi i organelama mrkih adipocita je hormonski regulisana, prije svega noradrenalinom. Mitohondrije, s jedne strane, imaju važnu ulogu u oblikovanju signalnih puteva kalcijuma, a kalcijum, s druge strane, u fiziološkim koncentracijama stimuliše mitohondrijalni metabolizam i povećava produkciju energije, dok u visokim koncentracijama indukuje apoptozu. Insulin je jedan od glavnih modulatora termogene funkcije mrkih adipocita, gdje stimuliše sintezu i deponovanje lipida u mrkim adipocitima, proliferaciju mrkih adipocita, angiogenezu i vazodilataciju. Pokazano je da insulin utiče na aktivnost elektron transportnog lanca, a i da mitohondrije pojačavaju insulinsku osjetljivost putem redoks regulacije proteina tirozin kinaze i insulinskog receptora. Cilj ove doktorske disertacije je rasvjetljavanje kako kalcijum, s jedne strane, a insulin, s druge strane, utiču na molekulske mehanizme koje su uključene u strukturno remodeliranje mitohondrija mrkih adipocita pacova. U eksperimentu su korišćeni Wistar pacovi, starosti dva mjeseca. U eksperimentu sa Ca-SANDOZ, životinje su podijeljene u dve grupe, jedna je pila Ca-SANDOZ rastvoren u vodi, a druga, kontrolna, česmensku vodu, tri dana. U eksperimentu sa insulinom, pacovi su podijeljeni u šest grupa, četiri grupe su primale nisku (0.4 IU) ili visoku (4 IU) dozu insulina i to jedan (akutno) ili tri dana (hronično). Ostale dvije grupe, kontrolne, su primale fiziološki rastvor akutno ili hronično. Trećeg dana tretmana Ca-SANDOZ, odnosno tri sata poslije posljednjeg tretmana insulinom, životinje su žrtvovane dekapitacijom, a interskapularni depo BAT je izolovan i pripremljen za biohemijske, mikroskopske i molekularno-biološke analize, i za izolaciju mitohondrijalnih frakcija. BAT i mitohondrijama bogata frakcija su analizirane Western Blot analizom, imunohistohemijskom, imunofluorescentnom i imunogold analizama, ultrastukturnim i stereološkim analizama, a analizirana je i aktivnost ATP sintaze...Essential role of brown adipose tissue is maintaining body temperature and energy homeostasis of organism. Exposure to cold and increased diet intake lead to stimulation, proliferation and differentiation of brown adipocytes, what is followed by remodeling of mitochondria, thermogenic organelles of brown adipocytes. In the context of structural remodeling, mitochondria show wide repertoire of morphological changes (e.g. mitochondria number in cell, mean diameter, fusion and fission of mitochondria, volume density of mitochondrial cristae) what is connected with abundance of components of electron transport chain (ETC), ATP synthase, UCP proteins - especially UCP1 protein, and other proteins of inner mitochondrial membrane. Calcium plays important role in thermogenesis, and its concentration in cytosol and organelles of brown adipocytes is hormone regulated, particularly by noradrenaline. Mitochondria has essential role in shaping calcium signaling pathways, and calcium, on other hand, stimulates mitochondrial metabolism and increases energy production, but in high concentrations induces apoptosis. Insulin is one of major modulators of thermogenic function of brown adipocytes, and stimulates synthesis and storage of lipids in brown adipocytes, proliferation of brown adipocytes, angiogenesis and vasodilatation. It is showed that insulin has effect on electron transport chain activity, but also mitochondria enhances insulin sensitivity via redox regulation of tyrosine kinases and insulin receptor. The aim of this doctoral dissertation is elucidation how calcium and insulin effect on molecular mechanisms involved in structural remodeling of mitochondria of rat brown adipocytes. Two-month-old male Wistar rats were used. In Ca-SANDOZ study, they were divided into two groups - Ca-SANDOZ drinking or tap water drinking for three days. In insulin experiment, rats were divided into six groups, where four groups treated with low (0.4 IU) or high (4 IU) dose of insulin, for one (acutely) or three (chronically) days. Another two groups served as control, treated with 0.9% saline solution for one or three days. On third day of Ca-SANDOZ treatment, or three hours after last administration of insulin, they were sacrificed by decapitation, and interscapular portion of BAT was isolated and prepared for biochemical, microscopic and molecular biology analyses, also for isolation of mitochondria-enriched BAT fraction. BAT and mitochondria-enriched fraction were analyzed by Western blot analysis, imunohistochemical, immunofluorescent, immunogold, ultrastructural and stereological analyses. Also, ATP synthase activity was measured..

Methods for studying the localization of mitochondrial complexes III and IV by immunofluorescent and immunogold microscopy

The localization of proteins within a cell is very important for studying protein colocalization and subsequently understanding protein-protein interactions at the subcellular level. Using mitochondrial protein localization as a model, we established methods to study the localization of electron transport chain complexes (ETCCs), specifically complexes III and IV, in brown adipose tissue (BAT) and mitochondria. Immunofluorescent and immunogold techniques were applied to BAT paraffin sections and thin Araldite sections of mitochondria-enriched fractions, respectively. Microscopic analysis clearly showed mitochondrial localization of complexes III and IV, as well their colocalization. In addition, 10 and 20 nm gold particles were capable of identifying the localization of complexes within mitochondrial cristae. The methods described in this study may be a beneficial addition to currently utilized methods for accurately identifying the localization of ETCCs, their colocalization with other proteins and their distribution inside the cell and cellular compartments. Lastly, this method can also be used to study the molecular architecture of BAT mitochondria by analyzing fixed and postfixed thin plastic sections with electron microscopy (EM). [Projekat Ministarstva nauke Republike Srbije, br. 173054 i br.173055

In vitro and in vivo antagonistic activity of new probiotic culture against Clostridium difficile and Clostridium perfringens

Abstract Background Genus Clostridium accompanies more than 200 known species and at least 30 among them are associated with human and animal diseases. At the moment, the treatment of clostridial infections is based on use of antibiotics. However, due to the European ban on the use of antibiotics in livestock production, novel therapeutic strategies for treatment of these hardly curable infections have been evaluated. Hence, in this study the antimicrobial effect of newly designed probiotic culture consisted of natural isolates Lactobacillus helveticus BGRA43, Lactobacillus fermentum BGHI14 and Streptococcus thermophilus BGVLJ1-44 against Clostridium difficile and Clostridium perfringens was analyzed. Results The probiotic culture showed strong in vitro antimicrobial effect on C. difficile (human clinical isolate). In addition, individual strains and the probiotic combination exhibited immunomodulatory activity. The probiotic combination significantly increased the proliferation of GALT lymphocytes. At the other hand, none of the bacterial treatments (individual strains and the combination) induced the production of proinflammatory cytokines IL-6 and IL-1β by intestinal epithelial cells, Caco-2. Interestingly, Caco-2 cells exposed to the probiotic combination produced significantly elevated amount of TGFβ pointing to potential protecting effect of the probiotic. In addition, the results of field trial on spontaneously infected goats revealed reduction of C. perfringens in goats (below the detection threshold) after the probiotic treatment. Conclusions The results of this study indicated that the novel probiotic deserves to be further investigated as a promising antimicrobial agent against C. difficile and C. perfringens

The interaction between SBA-15 derivative loaded with Ph3Sn(CH2)6OH and human melanoma A375 cell line: uptake and stem phenotype loss

Extraordinary progress in medicinal inorganic chemistry in the past few years led to the rational design of novel platinum compounds, as well as nonplatinum metal-based antitumor agents, including organotin compounds, whose activity is not based on unrepairable interaction with DNA. To overcome poor solubility and toxicity problems that limited the application of these compounds numerous delivering systems were used (Lila et al. in Biol Pharm Bull 37:206–211, 2014; Yue and Cao in Curr Cancer Drug Targets 16:480–488, 2016; Duan et al. in WIREs Nanomed Nanobiotechnol 8:776–791, 2016). Regarding high drug loading capacity, mesoporous silica nanoparticles like SBA-15 became more important for targeted drug delivery. In this study, cellular uptake and biological activities responsible for organotin(IV) compound Ph3Sn(CH2)6OH (Sn6) grafted into (3-chloropropyl)triethoxysilane functionalized SBA-15 (SBA-15p → SBA-15p|Sn6) were evaluated in human melanoma A375 cell line. Moreover, the influence of SBA-15p grafted with organotin(IV) compound on the stemness of A375 cell was tested. Given the fact that SBA-15p|Sn6 nanoparticles are nonspherical and relatively large, their internalization efficiently started even after 15 min with stable adhesion to the cell membrane. After only 2 h of incubation of A375 cells with SBA-15p|Sn6 passive fluid-phase uptake and macropinocytosis were observed. Inside of the cell, treatment with SBA-15p loaded with Sn6 promoted caspase-dependent apoptosis in parallel with senescence development. The subpopulation of cells expressing Schwann-like phenotype arose upon the treatment, while the signaling pathway responsible for maintenance of pluripotency and invasiveness, Wnt, Notch1, and Oct3/4 were modulated towards less aggressive signature. In summary, SBA-15p enhances the efficacy of free Sn6 compound through efficient uptake and well profiled intracellular response followed with decreased stem characteristics of highly invasive A375 melanoma cells

Promotion of Early Gut Colonization by Probiotic Intervention on Microbiota Diversity in Pregnant Sows

The aim of this work was to design a novel mixed probiotic culture for piglets and to evaluate its beneficial effect on the piglets’ gut health. The possible mechanisms of probiotic activity, such as adhesion, competitive pathogen exclusion and influence on gut microbiota diversity were determined. Mixed probiotic starter culture is composed of three thermophilic lactic acid bacteria (LAB) strains: Lactobacillus helveticus BGRA43, Lactobacillus fermentum BGHI14 and Streptococcus thermophilus BGVLJ1-44. The strains BGVLJ1-44 and BGRA43 showed good technological properties (fast milk curdling, strong proteolytic activity). In addition, the strain BGVLJ1-44 produces exopolysaccharide (EPS), BGHI14 is heterofermentative LAB strain with significant immunomodulatory effect, while the strain BGRA43 showed strong antimicrobial activity against different pathogens and exhibited significantly higher level of adhesion to Caco-2 cells comparing to other two strains. Both lactobacilli strains BGRA43 and BGHI14 (p < 0.05), as well as probiotic combination (p < 0.01) significantly reduced the adhesion of Escherichia coli ATCC25922 to Caco-2 cells, while the strains BGVLJ1-44 (p < 0.01) and BGRA43 (p < 0.05) significantly reduced adhesion of Salmonella 654/7E (veterinary isolate). The results of farm trial revealed that treatment of sows with new fermented dairy probiotic influenced the piglets’ gut colonization with beneficial bacteria and reduced the number of enterobacteriaceae in litters from some treated sows (no significant due to high variability among animals). Finally, this is the first study reporting that the treatment of sows with probiotic combination resulted in the improved microbiota diversity in neonatal piglets