miR-20b and miR-451a Are Involved in Gastric Carcinogenesis through the PI3K/AKT/mTOR Signaling Pathway: Data from Gastric Cancer Patients, Cell Lines and Ins-Gas Mouse Model

Gastric cancer (GC) is one of the most common and lethal gastrointestinal malignancies worldwide. Many studies have shown that development of GC and other malignancies is mainly driven by alterations of cellular signaling pathways. MicroRNAs (miRNAs) are small noncoding molecules that function as tumor-suppressors or oncogenes, playing an essential role in a variety of fundamental biological processes. In order to understand the functional relevance of miRNA dysregulation, studies analyzing their target genes are of major importance. Here, we chose to analyze two miRNAs, miR-20b and miR-451a, shown to be deregulated in many different malignancies, including GC. Deregulated expression of miR-20b and miR-451a was determined in GC cell lines and the INS-GAS mouse model. Using Western Blot and luciferase reporter assay we determined that miR-20b directly regulates expression of PTEN and TXNIP, and miR-451a: CAV1 and TSC1. Loss-of-function experiments revealed that down-regulation of miR-20b and up-regulation of miR-451a expression exhibits an anti-tumor effect in vitro (miR-20b: reduced viability, colony formation, increased apoptosis rate, and miR-451a: reduced colony forming ability). To summarize, the present study identified that expression of miR-20b and miR-451a are deregulated in vitro and in vivo and have a tumor suppressive role in GC through regulation of the PI3K/AKT/mTOR signaling pathway

AN ESSENTIAL OIL COMPOSITION OF THE NEEDLES OF SCOTS PINE (PINUS SYLVESTRIS L.) GROWING IN THE SOILS OF DIFFERENT TYPE

Extracts of pines have diverse physiological and pharmacological actions, its buds, needles and bark are used in phytotherapy.Geographical, seasonal, age-related, inter-population differencies of volatile compounds in the needles were defined. Wider surveys of the needle essential oil composition under the different type of soils are still scarse. Despite Scots pine is the prevailing tree in the forests of Lithuania, little is known about its essential oils. The aim of our study was to evaluate the essential oil composition in the needles of Scots pine (Pinus sylvestris L.) growing on arenosols (3 sites), luvisols (3 sites) and histosols (3 sites). Current-year and one-year-old needles of 8 pines in each site were sampled in July, 2005. Volatile components of the needles were extracted and analyzed by GC and GC/MS. Seventy one identified component made up 89.1-95.1 % of total oil content. The most predominant fraction was found to be monoterpenes (19.0-40.0 %), with pinene (6.1-26.1 %) and 3-carene (4.9-22.9 %) as the major constituents. Between the pine stands growing on the soils of different type statistically significant changes in percentage concentrations of monoterpens, sesquiterpens, oxysesquiterpens, diterpens were documented. For the pines growing on histosols and luvisols bigger amounts of shorter chain terpens and smaller amounts of longer chain terpens are produced when compared to pines on arenosols. Determined changes in the proportion of components of the essential oils in the needles of the trees growing on different types of soil may significantly modify medicinal value of the needles

Changes in the Essential Oil Composition in the Needles of Scots Pine ( Pinus sylvestris

Unfavorable anthropogenic factors, such as air pollution, lead to biochemical responses in trees. Changes in the amounts of secondary metabolites may be early indicators of invisible injuries. The aim of this study was to evaluate composition of the essential oils in the needles of Scots pine (Pinus sylvestris L.) growing in the areas affected by pollutant emissions of main factories in Lithuania: a nitrogen fertilizer factory (NFF), a cement factory (CF), and an oil refinery (OR). Totally, 14 pine stands were examined along transects from the factories (July 2005). Volatile components of the needles were extracted and analyzed by GC and GC/MS. Over 70 components of the essential oils were identified in current-year and 1-year-old needles

BIOMARKERS AMONG TERPENOIDS AND QUANTITY OF EPIPHYTIC MICROFLORA OF ARTEMISIA L. SPECIES NATIVE FOR LITHUANIA

Species belonging to Artemisia L. genus are rich in secondary metabolites (flavonoids, essential oils) and have been widely used in pharmacology and food industry. The concentration of the secondary metabolites depends on nutrition and developmental stage of the plant. Little is known about biology of Artemisia species (A. campestris L., A. vulgaris L. and A. absinthium L.) naturally growing in Lithuania. The aim of the work included evaluation of biomarkers among terpenoids in the plants, growing under different climatic and environmental conditions. Morphological properties of the plants were evaluated. For the quantity and chemical composition of the essential oils the following methods and techniques were used: gas chromatograph (HP 5890 II) with mass spectrometric detector (HP 5971), HPLC (chromatograph Waters 2690) equipped with VIS-UV detector (Waters 2487) and VIS-UV spectrophotometer (Genesys 5). The main biomarkers among terpenoids were determined (major specific compounds, with amount ³ 10 %): a- ir b -pinenes, sabinene, thujones, 1,8-cineole (for A. vulgaris), epoxi-ocimene (for A. absinthium), chrysanthenyl acetate (for A. vulgaris), trans-caryophyllene, germacrene D, trans-sabinyl acetate (for A. absinthium ), g-curcumene+acoradiene (for A. campestris). Synthesis of the secondary metabolites is influenced by nutrition and related to the quantity of the epiphytic microorganisms. According to the quantity of epiphytic microflora developmental differences were observed, also aboveground organ specificity was documented. No expressed differences were found between investigated species according to the amount of surface microorganisms. The differences in nitrogen nutrition got the best reflection in the ratio of the amounts of bacteria and fungi

Genetic diversity of Lithuanian populations of Juniperus communis L. in relation to abiotic and biotic factors

Juniperus communis L. is increasingly threatened in many parts of Europe, including the Baltic region. Our present study was aimed at evaluation of genetic diversity of J. communis populations of Lithuania. Fourteen selected populations differed in geography and habitats (coastal brown dunes covered with natural Scots pine forests, further referred as B; J. communis shrubs, F; transition mires and quaking bogs, D; subcontinental moss Scots pine forests, G; xero-thermophile fringes, E). Molecular variance was analyzed among populations (140 individuals, in total), employing 14 Inter Simple Sequence Repeat (ISSR) markers. Percentage of polymorphic loci differed between populations belonging to the different habitat type, being the highest for population representing habitat F (68.2%) and the lowest for populations D (42.9%). The data demonstrate the possibility to underestimate or overestimate this parameter if some habitat populations are bypassed. Means per population of Nei‘s gene diversity and Shannon‘s information indexes, were 0.158 and 0.239, respectively. Significant correlation between genetic and geographic distance of populations of J. communis was documented by Mantel test. Bayesian analysis of ISSR data has separated populations of Southern Lithuania from Northern part. Juniperus communis populations representing different habitats showed moderate interpopulation variance at ISSR loci. In ISSR-based dendrograms, individuals were correctly allocated to populations, even in case of populations growing in 0.5 km vicinity (habitats D and G). Among all populations the most distinct one was representing habitat of J. communis shrubs (F) protected by EUNIS. Principal coordinate analysis of weighted averages of Ellenberg’s indicator values (EIV) for herbaceous species, separated populations according to habitat type, while the same type analysis of ISSR data allowed distinguishing E habitat populations from populations of all other habitat type but G. Our study shows that on the local (Lithuanian) scale, molecular variance between populations of J. communis at ISSR loci might be related to habitat type. For evaluation and preservation of diversity of J. communis, different assessment methods should be included and management strategies should be directed in the way to retain variety of habitats encompassing both widely spread and less common ones

Caryophyllene Oxide-rich Essential Oils of Lithuanian Artemisia campestris ssp. campestris and Their Toxicity

International audienceThe chemical composition of the essential oils of aerial parts of Artemisia campestris ssp. campestris, collected from ten different locations in Lithuania is detailed in this paper. The major component in all the oils was caryophyllene oxide (8.5-38.8%), whereas compounds with the caryophyllane skeleton ranged from 10.2 to 44.5%. Other representative constituents were germacrene D (≤15.0%), humulene epoxide II (≤8.1%), β-ylangene (≤7.7%), spathulenol (≤6.8%), β-elemene (≤6.8%), β-caryophyllene (≤6.2%), junenol (≤6.1%) and α-or β-pinene (≤5.5%). Eighty-seven compounds were identified, comprising 73.6-92.3% of the oils. The chemical composition was highly variable depending on the sample location. Toxicity of A. campestris oils was determined using the brine shrimp (Artemia sp.) assay. LC 50 values ranging to 20 µg/mL were obtained for three of the oils after 24 hours of exposure. Data of this test revealed that A. campestris ssp. campestris essential oils with dominant caryophyllene oxide are notably toxic

RAPD analysis of genetic diversity among Lithuanian populations of Impatiens glandulifera

Abstract In some Asian countries, the seeds of Himalayan balsam or policeman's helmet, Impatiens glandulifera Royle are harvested as a food source. First introduced to Europe in 1838-1839, I. glandulifera quickly became a garden favourite and later on a prodigious weed. Nowadays I. glandulifera is highly invasive in almost whole Europe and occurs in various habitats. So far, little has been known about the genetic diversity of I. glandulifera in the Baltic region. The objective of this study was to evaluate the genetic variability of Lithuanian populations of I. glandulifera differing in geography or habitats by the randomly amplified polymorphic DNA (RAPD), using 8 primers. At the species level, all DNA band

Genetic diversity of Lithuanian populations of Juniperus communis L. in relation to abiotic and biotic factors

Juniperus communis L. is increasingly threatened in many parts of Europe, including the Baltic region. Our present study was aimed at evaluation of genetic diversity of J. communis populations of Lithuania. Fourteen selected populations differed in geography and habitats (coastal brown dunes covered with natural Scots pine forests, further referred as B; J. communis shrubs, F; transition mires and quaking bogs, D; subcontinental moss Scots pine forests, G; xero-thermophile fringes, E). Molecular variance was analyzed among populations (140 individuals, in total), employing 14 Inter Simple Sequence Repeat (ISSR) markers. Percentage of polymorphic loci differed between populations belonging to the different habitat type, being the highest for population representing habitat F (68.2%) and the lowest for populations D (42.9%). The data demonstrate the possibility to underestimate or overestimate this parameter if some habitat populations are bypassed. Means per population of Nei‘s gene diversity and Shannon‘s information indexes, were 0.158 and 0.239, respectively. Significant correlation between genetic and geographic distance of populations of J. communis was documented by Mantel test. Bayesian analysis of ISSR data has separated populations of Southern Lithuania from Northern part. Juniperus communis populations representing different habitats showed moderate interpopulation variance at ISSR loci. In ISSR-based dendrograms, individuals were correctly allocated to populations, even in case of populations growing in 0.5 km vicinity (habitats D and G). Among all populations the most distinct one was representing habitat of J. communis shrubs (F) protected by EUNIS. Principal coordinate analysis of weighted averages of Ellenberg’s indicator values (EIV) for herbaceous species, separated populations according to habitat type, while the same type analysis of ISSR data allowed distinguishing E habitat populations from populations of all other habitat type but G. Our study shows that on the local (Lithuanian) scale, molecular variance between populations of J. communis at ISSR loci might be related to habitat type. For evaluation and preservation of diversity of J. communis, different assessment methods should be included and management strategies should be directed in the way to retain variety of habitats encompassing both widely spread and less common ones

Thrombosis Related ABO, F5, MTHFR, and FGG Gene Polymorphisms in Morbidly Obese Patients

Objective. Obesity is a well-known risk factor for thrombotic complications. The aim of the present study was to determine the frequency of thrombosis related ABO, F5, MTHFR, and FGG gene polymorphisms in morbidly obese patients and compare them with the group of nonobese individuals. Methods. Gene polymorphisms were analyzed in 320 morbidly obese patients (BMI > 40 kg/m2) and 303 control individuals (BMI T (rs505922), F5 C>G (rs6427196), MTHFR C>T (rs1801133), and FGG C>T (rs6536024) SNPs were genotyped by RT-PCR. Results. We observed a tendency for MTHFR rs1801133 TT genotype to be linked with morbid obesity when compared to CC genotype; however, the difference did not reach the significant P value (OR 1.84, 95% CI 0.83–4.05, P=0.129). Overall, the genotypes and alleles of rs505922, rs6427196, rs1801133, and rs6536024 SNPs had similar distribution between morbidly obese and nonobese control individuals. Distribution of height and weight means among individuals carrying different rs505922, rs6427196, rs1801133, and rs6536024 genotypes did not differ significantly. Conclusions. Gene polymorphisms ABO C>T (rs505922), F5 C>G (rs6427196), MTHFR C>T (rs1801133), and FGG C>T (rs6536024) were not associated with height, weight, or morbid obesity among European subjects