Procter & Gamble (United Kingdom)
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Ponašanje NMRI miševa u uzdignutom krstastom lavirintu prenatalno izloženih kombinaciji valproinske kiseline I levetiracetama (1:1)
No full textUvod: Gestacija kao entitet, zahteva kontinuirano praćenje, a posebnu pažnju i nadzor u slučaju primene farmakoterapije u tretmanu epilepsija. Lečenje epilepsije tokom gestacije je kompleksan i izazovan zadatak. Politerapija se uvodi u slučaju pojave epi- napada usled neadekvatnog odgovora na određeni antiepileptik tokom trudnoće. Na osnovu istraživanja (Sarhan et al., 2015; Dudra- Jastrzebska, 2009), nije zabeležen neželjeni uticaj na potomstvo primenom valproinske kiseline (VPK) i levetiracetama (LEV) u odnosu 1:1, pored aditivnog efekta. Cilj ove studije je da se ispita ponašanje miševa, čije su majke bile kontinuirano izložene kombinaciji VPK i LEV (1:1) tokom celokupne gestacije.
Metode: U ovoj studiji korišćene su odrasle ženke miša NMRI soja, stare 8 nedelja. Formirane su dve grupe životinja: prva grupa (8 ženki) tretirana je kombinacijom LEV u dozi od 211 mg/kg/dan (LEV-1000) i VPK u dozi od 200 mg/kg/dan (VPK-1000) i kontrolna grupa (10 ženki) koje su dobile odgovarajuću zapreminu fiziološkog rastvora (CTR). Primenjene doze odgovaraju ljudskim dozama od 1000 mg/dan za oba antiepileptika. Sve životinje su tretirane subkutanom injekcijom, u kožu na zadnjem delu vrata dva puta dnevno. Tretman je započet kada su mužjaci i ženke upareni i nastavljen je tokom celog perioda gestacije dok se ženke nisu okotile. Potom, svaka ženka sa svojim leglom je smeštena posebno. Životinje su razdvojene po polu 20-og postnatalnog dana(PND). Ponašanje je testirano na PND 37 u testu uzdignutog krstastog lavirinta.
Diskusija: Potomstvo izloženo kombinaciji VPA+LEV favorizuje ponašanje procene rizika u testu uzdignutog krstatsog lavirinta. Naime, mužjaci i ženke u grupi prenatalno tretiranih kombinacijom VPK i LEV su ispoljili statistički značajanu razliku analizom kvantitativnih parametara u odnosu na kontrolu u testu uzdignutog krstastog lavirinta.
Zaključak: Imajući u vidu korelaciju između promena u ponašanju jedinki prenatalno izloženih kombinaciji VPK I LEV i nekih specifičnih karakteristika adolescencije, nameće se neophodnost dugotrajnog praćenja dece prenatalno izložene antiepileptičkoj.Sokić D, editor. Zbornik sažetaka: XX kongres Društva neurologa Srbije sa međunarodnim učešćem; 2024 Nov 14-16; Belgrade, Serbia. Beograd: Društvo neurologa Srbije; 2025. p. 114
The role of lung and gut microbiota in the immune response to Aspergillus fumigatus in Dark Agouti and Albino Oxford rat strains
No full textBakterijska mikrobiota prisutna u organima uključena je u fiziološke (održavanje homeostaze) i patofiziološke procese (utiče na ishod bolesti) kako lokalno, tako i u udaljenim organima. Uticaj mikrobiote creva na imunske reakcije u plućima je dobro okarakterisan, ali malo je podataka o uticaju inflamacije u plućima na mikrobiotu u samim plućima, ali i u crevima. Upotrebom metode sekvenciranja najnovije generacije ispitane su promene u bakterijskoj mikrobioti pluća i creva kod jedinki dva soja pacova Dark Agouti (DA) i Albino Oxford (AO) (za koje je pokazano da ispoljavaju kvantitativno različit imunski odgovor na isti stimulus) kod kojih je infekcija pluća izazvana sa A. fumigatus. Bakterijska mikrobiota pluća zdravih jedinki ova dva soja se ne razlikuje, ali su u crevima primećene razlike u sastavu. Kod oba soja dolazi do disbioze u plućima nakon infekcije sa A. fumigatus, koja je verovatno posledica imunskog odgovora na gljivu (povećanje IFN-γ, IL-17 i MPO kod oba soja) i oksidativnog stresa (kod DA pacova). Tokom infekcije pluća javlja se inflamacija i oksidativni stres i u crevima kod oba soja, ali disbioza u crevima je primećena samo kod DA pacova. Narušavanje ravnoteže bakterijske mikrobiote samo u crevima (primenom antibiotika) dovodi do povećanja osetljivosti oba soja na infekciju sa A. fumigatus. Kod jedinki koje su pre infekcije pile antibiotike zapaža se veći stepen aktivacije ćelija limfnih čvorova pluća (Th1 i Treg ćelija kod DA, Th17 ćelija kod AO) i inflamacija u tkivu pluća (povećanje IL-1β i IL-17) samo kod DA. Ovi rezultati doprinose saznanjima o ulozi mikrobiote creva i pluća tokom infekcije pluća i ukazuju na potencijalne mehanizme dvosmerne komunikacije između ovih organa.Bacterial microbiota is involved in physiological (maintenance of homeostasis) and pathophysiological processes (associated with disease outcome) both locally and in distant organs. The influence of gut microbiota on immune responses in the lungs is well characterized, but there is little data on the effect of lung inflammation on both lung and gut microbiota. Using the Next generation sequencing method, changes in the bacterial microbiota of the lungs and gut were examined in two rat strains, Dark Agouti (DA) and Albino Oxford (AO) (shown to exhibit a quantitatively different immune response to the same stimulus) in which lung infection was caused by the fungus A. fumigatus. The lung microbiota between healthy DA and AO rat strains does not differ, but differences in the gut microbiota composition were observed. After infection with A. fumigatus dysbiosis was noted in the lungs of both strains, which is probably the result of immune response to the fungus (increased IFN-γ, IL-17 and MPO in both strains) and oxidative stress (in DA rats). During lung infection, inflammation and oxidative stress were noted in the gut of both strains, but dysbiosis only in DA rats. Gut microbiota disturbance limited to the intestines only (using antibiotics) in both strains, leads to an increased risk of infection with A. fumigatus. Only in DA animals who received antibiotics before infection, a higher lung lymph node cell activation (Th1 and Treg cells in DA, Th17 cells in AO) and lung tissue inflammation (an increase of IL-1β and IL-17) were observed. These results contribute to knowledge about the role of gut and lung microbiota during lung infection and indicate potential mechanisms of bidirectional communication between these organs
Trehalose differently regulates mitochondrial-oxidative stress damage in 6-OHDA and MPP+ - induced toxicity in neuroblastoma SH-SY5Y cells by regulating p38, JNK and AMPK/Akt/mTOR signaling pathways
No full text6-OHDA and MPP+ are the most common neurotoxins used in experimental models
of Parkinson's disease. Neurotoxicity induced by 6-OHDA and MPP+ is mediated
by mitochondrial dysfunction and oxidative stress leading to cell death. In this study,
we investigated the effects of natural disaccharide trehalose on 6-OHDA- and
MPP+-induced toxicity in SH-SY5Y human neuroblastoma cells.
Methods - Cell viability, flow cytometry, immunoblot, transmission electron
microscopy.
As shown by flow cytometry of mitosoxRed-stained cells and immunoblot analyses,
trehalose decreases mitochondrial superoxide production and cleavage/activation of
caspase 3 in 6-OHDA-treated cells. In MPP+-exposed cells, trehalose increased the
cleavage/activation of caspase 3 and the intensity of mitosoxRed fluorescence.
Accordingly, TEM analysis of 6- OHDA-exposed cells pre-treated with trehalose
showed mitochondria with typical ultrastructure in an electron-dense matrix without
signs of damage. When MPP+ was combined with trehalose, TEM showed large,
swollen mitochondria with fragmentation and disorganization of the cristae in a
markedly electron-lucid matrix. In addition, immunoblot showed that trehalose
significantly decreased the phosphorylation of p38 and Akt upon treatment with 6
OHDA, while the activity of these molecules increased in MPP+ exposed cells. In
addition, trehalose decreased the phosphorylation of AMPK and increased the
phosphorylation of S6 kinase in both 6-OHDA- and MPP+-treated cells, whereas the
activation of JNK was decreased by trehalose only in 6-OHDA treatment. In contrast
to trehalose, the antioxidant N-acetylcysteine (NAC) reduced the phosphorylation
of p38 both in 6-OHDA and MPP+. Moreover, as shown by MTT and LDH assays
pharmacological inhibition of JNK SP600125 fully reproduced and potentiated the
protective effect of trehalose. Immunoblot analysis showed that SP decreased
phosphorylation of p38 same as trehalose, while the phosphorylation of Akt was additionally decreased in 6- OHDA–exposed cells pretreated with trehalose,
suggesting the possible mechanism of action in trehalose-mediated protection
against 6-OHDA-induced apoptosis.
In conclusion, trehalose differently regulates mitochondrial oxidative stress damage
induced by two different neurotoxins 6-OHDA and MPP+ , suggesting further
research on the regulation of signaling molecules and their subcellular localization
to preserve mitochondrial function in neurodegeneration.Dubljević O, Marjanović A, editors. Book of Abstracts: Belgrade Neuroscience Next Hub 2024 with international participation; 2024 May 24-25; Belgrade, Serbia. Belgrade: University of Belgrade, Faculty of Medicine; Serbian Neuroscience Society; 2024. p. 64-5
The Influence of Dietary Cadmium on Changes in the Midgut Mass Related to the Mass of Gypsy Moth Larvae
No full textCadmium pollution is becoming an increasing problem, especially in parts of the world that have developed industries. To consider the potentially harmful effects of cadmium, we need to examine changes at all different levels of biological organization. The main goal of this study was to detect a possible change in the percentage of midgut mass relative to larval mass (PMM) and determine the plasticity of this trait and the correlations between midgut enzymes and PMM under stress conditions. Fourth-instar larvae were exposed to acute and chronic effects of two cadmium concentrations, 10 and 30 μg Cd/g dry food, as well as a three-day recovery from chronic treatments. PMM is also an indirect indicator of food consumption and was found to be significantly reduced compared to control in both acute effects and chronic treatment at 30 μg and its three-day recovery. The PMM reduction during acute treatments is a consequence of cadmium action, while in chronic treatment, the genetic factor (egg mass) plays a crucial role in the change of PMM. According to the index of plasticity, distinct phenotypes were not produced. Significant correlations were shown between PMM and trypsin (Tryp) and leucine aminopeptidases (LAP) at acute and chronic treatment with higher cadmium concentrations, while significant correlations between proteases and PMM were detected at lower metal concentrations (Acute10 and Chronic10 and 30 μg Cd/g dry food). In contrast to chronic treatment, egg masses respond more uniformly by reducing PMM during the short-term effect of cadmium. Finally, we can conclude that, as an addition to biochemical and molecular research, PMM can be used for studying the cadmium effects to gain a better insight into the state of the organism under stress conditions
Chemical Analysis and Investigation of Antimicrobial and Antibiofilm Activities of Prangos trifida (Apiaceae)
No full textPlants of the genus Prangos are intensively investigated as potential new sources of bioactive isolated products. In this work, the chemical composition of volatile constituents (essential oils and headspace volatiles) and dichloromethane extracts, as well as antimicrobial and antibiofilm activities of essential oils and MFDEs (methanol fractions of dichloromethane extracts) of Prangos trifida from Serbia, were investigated. Volatiles of roots, leaves, stems and fruits, and fatty acids and phytosterols in dichloromethane extracts of roots and fruits were analyzed by GC-FID-MS, whereas coumarins in MFDEs by LC–MS and some isolated coumarins by 1H-NMR. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations/minimum fungicidal concentrations (MBCs/MFCs) of essential oils and MFDEs were determined against 13 microorganisms. Antibiofilm activity was assessed against four microorganisms. Additionally, congo red and ergosterol binding assays were conducted to elucidate selected mechanisms of antibiofilm action in the case of Candida albicans. Total of 52 volatile constituents, 16 fatty acids, eight phytosterols and 10 coumarins were identified. Essential oils demonstrated significant activity, surpassing that of commercial food preservatives, against six tested molds from the Aspergillus, Penicillium and Trichoderma genera, as well as against bacteria Staphylococcus aureus and Bacillus cereus. Most of the oils strongly inhibited the formation of biofilms by S. aureus, Listeria monocytogenes and Escherichia coli. MFDEs exhibited noteworthy effects against B. cereus and the tested Aspergillus species, particularly A. niger, and significantly inhibited C. albicans biofilm formation. This inhibition was linked to a marked reduction in exopolysaccharide production, while antifungal mechanisms associated with ergosterol remained unaffected
Biological activity of two coniferous essential oils against spongy moth larvae and Phytophthora root pathogens
No full textWe provide one excel (with five sheets) and 5 csv files. The data in the excel file is the same as the one provided in the 5 csv files. Data file named 'raw data.xlsx' is consisted of 5 sheets containing raw data on all analyzed traits in the manuscript for the spongy moth and Phytophthora sp. The first sheet named 'RDC' contains data on relative deterrence coefficients of spongy moth larvae exposed to three different concentrations of Calocedrus decurrens and Cupressus arizonica essential oils. The second sheet named 'ADC' contains data on absolute deterrence coefficients of spongy moth larvae exposed to three different concentrations of Calocedrus decurrens and Cupressus arizonica essential oils. The third sheet named 'RCR' contains data on the relative consumption rate of spongy moth larvae exposed to three different concentrations of Calocedrus decurrens and Cupressus arizonica essential oils, and control treatment. The forth sheet named ‘RGR’ contains data on relative growth rate of gypsy moth larvae exposed to three different concentrations of Calocedrus decurrens and Cupressus arizonica essential oils, and control treatment. The fifth sheet named ‘Ir’ contains data on the antimicrobial activity of three concentrations of Calocedrus decurrens and Cupressus arizonica essential oils against Phytophthora × cambivora, P. plurivora and P. quercina expressed as inhibition rate of mycelial growth
Anoikis and cancer cell differentiation: novel modes of shikonin derivatives anticancer action in vitro
No full textBackground: Shikonin is a naturally occurring naphthoquinone found in the roots of several genera of the Boraginaceae family, widely known for its numerous biological activities, such as antiinflammatory, antioxidant, antimicrobial and anticancer. In this study, the antitumor effect of six naphthoquinones isolated from the roots of Onosma visianii was evaluated using two cell lines, mouse melanoma B16 and highly aggressive rat glioma cell line C6.
Methods and results: All examined shikonins dose-dependently decreased the viability of tested cells, with compounds 5 and 6 being the most potent ones and hence subjected to further analysis. The diminished viability of B16 melanoma cells was in correlation with detected caspase-mediated apoptosis. Importantly, observed altered cell morphology along with the loss of dividing potential upon exposure to both shikonins implied reprogram of B16 cell phenotype. Elevated expression of myelin basic protein indicated the acquirement of Schwann‐like cell phenotype, while detected autophagy might be connected to this phenomenon. On the contrary, upon exposure to both agents, C6 cells underwent specific cell death—anoikis, provoked by detachment from the extracellular matrix and compromised integrin signaling. Oppositely to compound 5, compound 6 realized anoikis in a caspase-independent manner and under sustained ERK1/2 activation, indicating the deviation from standard proanoikis signaling.
Conclusions: Herein, we have pointed out the diversity and novelty in the mode of action of shikonin derivatives depending on the tumor cell features, which represents a good platform for new investigations of these promising natural compounds
Rapid and reliable method for identification of three medium‑sized horseshoe bat species in Europe
No full textAccurate species identification is crucial for ecological research and effective wildlife management. Advances in molecular genetic tools enable this, even for cryptic species complexes that are often morphologically confusing or indistinguishable. However, the costs of these methods and sequencing remain prohibitive for many researchers, particularly in less developed regions. The aim of this study was to test whether ISSR-PCR markers can be used to distinguish three similar horseshoe bat species that are often misidentified and occur in sympatry in Serbia. Samples from 64 Rhinolophus euryale, R. blasii, and
R. mehelyi bats were identified using this ISSR-PCR approach, and species identifications were additionally confirmed by sequencing D-loop fragment of mitochondrial DNA. ISSR-PCR yielded species-specific band patterns on the agarose gel that allowed the differentiation of three medium-sized horseshoe bat species. This approach does not require sequencing, making it a quick and inexpensive tool for the genetic identification of these species, and complements already existing methods. There is potential to scale up this method to other cryptic species complexes, reducing misidentifications that lead to inaccurate population trend assessments and have knock-on effects on our ability to monitor and conserve wildlife, especially rare and endangered species
Cytokinin deficiency confers enhanced tolerance to mild, but decreased tolerance to severe salinity stress in in vitro grown potato
No full textCytokinin (CK) is a plant hormone that plays crucial roles in regulating plant growth and development. CK-deficient plants are widely used as model systems for investigating the numerous physiological roles of CK. Since it was previously shown that transgenic or mutant CK-deficient Arabidopsis and Centaurium plants show superior tolerance to salinity, we examined the tolerance of three CK-deficient potato lines overexpressing the Arabidopsis thaliana CYTOKININ OXIDASE/DEHYDROGENASE2 (AtCKX2) gene to 50 mM, 100 mM, 150 mM, and 200 mM NaCl applied in vitro. Quantification of visible salinity injury, rooting and acclimatization efficiency, shoot growth, water saturation deficit, and chlorophyll content confirmed that the CK-deficient potato plants were more tolerant to low (50 mM) and moderate (100 mM) NaCl concentrations, but exhibited increased sensitivity to severe salinity stress (150 and 200 mM NaCl) compared to nontransformed control plants. These findings were corroborated by the data distribution patterns according to principal component analysis. Quantification of the activity of superoxide dismutases, peroxidases, and catalases revealed an impaired ability of AtCKX2-transgenic lines to upregulate the activity of antioxidant enzymes in response to salinity, which might contribute to the enhanced sensitivity of these potato lines to severe salt stress. Our results add complexity to the existing knowledge on the regulation of salinity tolerance by CK, as we show for the first time that CK-deficient plants can exhibit reduced rather than increased tolerance to severe salt stress
Dual Role of Alchemilla vulgaris L. Extract in Breast Cancer Regression: Reestablishment of Effective Immune Response
No full textEthnomedicinal records have long mentioned the historical usage of Alchemilla vulgaris L. in folk medicine, particularly for the treatment of gynecological issues. Building on this ethnomedicinal knowledge regarding female illnesses, the aim of this research was to evaluate the impact of ethanolic extract of A. vulgaris on mouse breast cancer cells (4T1) in vitro and in vivo, in addition to its effect on the immune compartment in the tumor microenvironment. Behind viability decrease of 4T1 cells induced by treatment with A. vulgaris extract was strong inhibition of cell proliferation accompanied by caspase-dependent apoptosis and autophagic cell death. Observed changes in 4T1 cell culture after treatment were well orchestrated and led to a reduction in metastatic potential through weakened adhesion, invasion, migration, and colony-forming abilities in vitro. Enhanced intracellular production of reactive oxygen and nitrogen species promoted by the treatment might interfere with all the observed effects. Apart from the direct effect on tumor cells, the A. vulgaris extract significantly reduced tumor growth in the solid orthotropic mammary carcinoma model through restitution of efficient local and systemic immune response reflected in enhanced antigen-presenting potential of dendritic cells (DCs) as well as the extent and activity of effector T cells