Volume 11, Issue 1

Camelid’ s heavy-chain antibody (HCAb) consists of only two heavy chains and lacks the two light chains together with the CH1 domain usually found in conventional immunoglobulins. A recombinant single antigen-binding entity, named VHH (or Nanobody®) was generated by reengineering the variable domains from HCAb. This study focuses on the detection of camelid´s immunoglobulins as well as their derivative nanobodies using a universal anti-camel antibody produced in rabbit (rIgG). Starting from a crude rabbit serum, a standard stock of rIgG (1 mg/ml) was prepared after purification by affinity chromatography using protein-A column. As expected, rIgG was able to detect camel antibodies in ELISA and immunoblotting, and its reactivity was equal against all different camel IgG subclasses, which were purified from serum by differential affinity chromatography on protein-G and -A. Interestingly, rIgG also recognized nanobodies since they were originally part of camel HCAbs, providing an alternative method to detect the corpus of these recombinant proteins rather than targeting their artificial tags. These data suggest that the anti-camel rIgG described here could be efficiently applied at different stages of nanobody technology, including the quantitation of the issued nanobodies and their detection when bound to target antigens.

The endemic Verbascum olympicum has characteristics that allows it to live in degraded areas of Uludağ Mountain, Turkey and therefore known as a ruderal species. In this study, V. olympicum seeds collected from Uludağ Mountain were grown in the Hoagland nutrient solution, under hydroponic conditions. The activity of antioxidative enzymes (superoxide dismutase, SOD; catalase, CAT; ascorbate peroxidase, APX) were examined to demonstrate the role of antioxidative mechanism in the seedlings exposed to different Cu concentrations (0, 50, 250, 500 μM) for seven days. Also, certain growth parameters (such as the water content, biomass production, soluble protein), the level of lipid peroxidation, cell membrane injury and permeability were investigated. As a result, some toxic effects are observed following the application of 500 μM Cu, after the seedlings growing in 50 and 250 mM Cu concentrations showed high resistance and survived in hydroponic conditions. Our findings provide information about the resistance of V. Olympicum seedlings to oxidative stress caused by excessive Cu concentrations.

Radiotherapy is commonly used to kill malignant cells, but it can significantly deplete hematopoietic and splenic erythroblasts. Radioprotective agents are therefore very important in clinical radiotherapy. We examined the effect of poly-herbal EMSA ERITIN on immunological responses when administered to sublethally irradiated mice with the aim of highlighting promotes erythroid lineages and lymphocytes migration in irradiated mice with the parameter are TER119 + CD123 + in bone marrow and SDF-1 in bone marrow and spleen organ. Normal BALB/c mice were sublethally irradiated with 600 rad. EMSA ERITIN was administered orally at different doses:(1.04, 3.125 and 9.375 mg/g body weight) for 15 days. On day 16 erythroid lineages (TER-119 + CD123 + ) were observed in bone marrow and lymphocytes migration by the production of SDF-1 in spleen and bone marrow. Lymphocytes migration was indicated by the production of SDF-1 in spleen and bone marrow using flow cytometry analysis. EMSA ERITIN increased the generation of erythroid lineage cells marked by TER119 + CD123 + and promoted lymphocyte migration by increasing SDF-1 production in bone marrow and spleen. EMSA ERITIN appears to be a powerful medicinal herb with potential as a food supplement to normalize homeostasis and erythropoiesis after radiation.

Cutin hydrolase (EC 3.1.1.74), an extracellular polyesterase found in pollens, bacteria and fungi, is an efficient catalyst that exhibits hydrolytic activity on a variety of water-soluble esters, synthetic fibers, plastics and triglycerides. Thus, cutinase can be used in various applications such as ester synthesis, bio-scouring, food and detergent industries. Ancut2 is one of five genes encoding cutinases present in the Aspergillus niger ATCC 10574 genome. The cDNA of Ancut2 comprising of an open reading frame of 816 bp encoding a protein of 271 amino acid residues, was isolated and expressed in Pichia pastoris . The partially purified recombinant cutinase exhibited a molecular mass of approximately 40 kDa. The enzyme showed highest activity at 40°C with a preference for acidic pH (5.0-6.0). AnCUT2 showed hydrolytic activity towards various p -nitrophenyl esters with preference towards shorter chain esters such as p -nitrophenyl butyrate (C4). Scanning Electron Microscopy demonstrated that AnCUT2 was capable of modifying surfaces of synthetic polycaprolactone and polyethylene terephthalate plastics. The properties of this enzyme suggest that it may be applied in synthetic fiber modification and fruit processing industries.

Staphylococcus aureus is the major causative agent of mastitis among dairy animals as it causes intramammary gland infection. Due to antibiotic resistance and contamination of antibiotics in the milk of diseased animals; alternative therapeutic agents are required to cure mastitis. Lytic bacteriophages and their gene products can be potential therapeutic agents against bacteria as they are host specific and less harmful than antibiotics. In this study, Staphylococcus aureus were isolated from milk samples of the infected animals and identified biochemically. SA phage was isolated from sewage water showing lytic activity against Staphylococcus aureus isolates. The highest lytic activity of bacteriophages was observed at 37°C and pH 7, and the most suitable storage condition was at 4°C. SA phage efficiently reduced bacterial growth in the bacterial reduction assay. The characterization and bacterial growth reduction activity of the bacteriophages against Staphylococcus aureus signifies their underlying potential of phage therapy against mastitis.

MafB is a member of bZip transcription factors that share similar basic region/leucine zipper DNA binding motifs and N-terminal activation domains. It is well known that MafB is highly expressed in macrophages and promotes differentiation of myeloid progenitors into macrophage. However, little is known about its function in dendritic cells. Here, we report that MafB, as a target of miR-155, which had been reported to be required for dendritic cell maturation and function, regulated dendritic cell maturation. MafB and miR-155were reversely correlated during DC maturation induced by LPS and forced expression of miR-155 reduced MafB expression. The luciferase reporter assay showed that MafB 3’UTR was directly targeted bymiR-155. In addition, knockdown of MafB promoted the phenotypic maturation of DC2.4 cells. Forced expression of MafB could significantly attenuate the phenotypic maturation of DC2.4 cells caused by overexpression of miR-155. Overall, our data demonstrates that MafB, inhibited by miR-155, was a negative regulator of DC maturation.

A protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 10 6 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N 6 -benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

Light quality is thought to affect the growth and development of plants. We examined how light influences the growth and content of some chemical compounds in dill ( Anethum graveolens L .). The plants were grown under different light quality. The share of orange and green light in the spectrum was constant and amounted to 10% for either colour. In the first combination (A, 70/10), there was 70% of red light and 10% of blue light. Other combinations had the following proportions: B 60/20, C 50/30, D 40/40 and E 30/50 of red and blue light. The PPFD was about 155 μmol m -2 s -1 . Blue light inhibited the elongation growth as well as leaf area. It had positive influence on the accumulation of dry mass, glucose and fructose in the herb. In the combinations with higher percentage of red light the plants were characterised by higher content of essential oils, macronutrients and zinc. To sum up, we can say that the proportion of red and blue light has significant influence on the morphological qualities, chemical composition and dynamics of photosynthesis in these plants. On the other hand, the selection of spectral composition of LEDs will depend on the result we want to achieve.

The activity of purified Fe 3+ containing horseradish peroxidase (HRP) was recorded while the enzyme was exposed to two low frequency pulsed electromagnetic fields (ELF-EMFs) with different frequencies: 50 Hz/2.7 mT and 100 Hz/5.5 mT. No statistically significant difference in the Michaelis constant ( K m ) values between the control enzyme and the enzyme exposed to any of the two ELF-EMF used could be detected (53 ± 11 μM vs 67 ± 10 μM vs 42 ± 11 μM respectively), indicating that upon short exposure times, there are no large structural alterations of the enzyme that affect the substrate affinity. A 50 Hz/2.7 mT EMF did however, cause a significant decrease in the maximum rate ( V max ) value (from 3.31 ± 0.17 μmoles/min to 1.6 ± 0.19 μmoles/min, p <0.03) and a drop of the catalytic efficiency to less than half (from 0.66 ± 0.03 s -1 to 0.32 ± 0.04 s -1 , p <0.05). These observations are the first experimental proofs that support the previously postulated mechanism of coupling between ELF-EMF and living systems based on a resonant frequency specific for a given ion molecule.

The effects of cold hardening of cereals on their cross-tolerance to treatments leading to oxidative stress were investigated. Long-term exposure to low non-freezing temperatures provided partial protection to wheat and barley plants from the damage caused by paraquat and hydrogen peroxide treatments. It also conferred resistance in two barley cultivars to the necrotic symptoms and growth of the fungal phytopathogen Pyrenophora teres f. teres . Pathogen-induced oxidative burst was also reduced in cold hardened plants. The possible roles of host-derived redox factors and other signaling components in the observed forms of cereal cross-tolerance are discussed.

Oculocutaneous albinism (OCA) is a genetic disorder of melanin synthesis that results in hypopigmentation in hair, skin and eyes. OCA has been reported in individuals from all ethnic backgrounds but it is more common among those with Europeans ancestry. OCA is heterogeneous group of disorders and seven types of OCA are caused by mutations in TYR (OCA1), OCA2 (OCA2), TYRP1 (OCA3), SLC45A2 (OCA4), SLC24A5 (OCA6) and C10oRF11 (OCA7) genes. However, MC1R gene variants have been reported that modify OCA2 phenotype but the knowledge about the function of MC1R gene in melanogenesis, and genotype-phenotype association, in case of OCA, is limited. In this review article we present a comprehensive description of classification of OCA, role of MSH-R in melanin synthesis, the sequence variations in MC1R and their association with OCA. This review will enhance our understanding of MC1R gene variants involved in human OCA2 phenotype.

In the present paper, the potential of canine articular cartilage-derived cells (cACCs) for chondrogenic differentiation was evaluated. The effectiveness of cACCs’ lineage commitment was analyzed after 14 days of culture in chondorgenic and non-chondrogenic conditions. Formation of proteoglycan-rich extracellular matrix was assessed using histochemical staining – Alcian Blue and Safranin-O, while elemental composition was determined by means of SEM-EDX. Additionally, ultrastructure of cACCs was evaluated using TEM. The expression of genes involved in chondrogenesis was monitored with quantitative Real Time PCR. Results obtained indicate that the potential of cACCs for cartilagous extracellular matrix formation may be maintained only in chondrogenic cultures. The formation of specific chondro-nodules was not observed in a non-chondrogenic culture environment. The analysis of cACCs’ ultrastructure, both in non-chondrogenic and chondrogenic cultures, revealed well-developed rough endoplasmatic reticulum and presence of mitochondria. The cACCs in chondrogenic medium shed an increased number of microvesicles. Furthermore, it was shown that the extracellular matrix of cACCs in chondrogenic cultures is rich in potassium and molybdenum. Additionally, it was determined that gene expression of collagen type II, aggrecan and SOX-9 was significantly increased during chondrogenic differentiation of cACCs. Results obtained indicate that the culture environment may significantly influence the cartilage phenotype of cACCs during long term culture.

In Leishmania species, protein disulfide isomerase (PDI) is an essential enzyme that catalyzes thiol-disulfide interchange. The present work describes the isolation, cloning, sequencing and expression of the pdI-2 gene. Initially, the gene was amplified from L. tropica genomic DNA by PCR using specific primers before cloning into the expression vector pET-15b. The construct pET/ pdI-2 was transformed into BL21(DE3) cells and induced for the protein expression. SDS-PAGE and western blot analysis showed that the expressed protein is about 51 kDa. Cloned gene sequence analysis revealed that the deduced amino acid sequence showed significant homology with those of several parasites PDIs. Finally, recombinant protein was purified with a metal-chelating affinity column. The putative protein was confirmed as a thiol - disulfide oxidoreductase by detecting its activity in an oxidoreductase assay. Assay result of assay suggested that the PDI-2 protein is required for both oxidation and reduction of disulfide bonds in vitro . Antibodies reactive with this 51 kDa protein were detected by Western blot analysis in sera from human infected with L. tropica . This work describes for the first time the enzymatic activity of recombinant L. tropica PDI-2 protein and suggests a role for this protein as an antigen for the detection of leishmaniasis infection.

There is a general decline of grasslands across Europe due to habitat loss and degradation. Ensuring plant dispersal thus becomes a key process for preserving grassland patches in all scales. We examined diaspore dispersal by sheep epizoochory in the pastures of the North Adriatic Karst (NW Slovenia) and determined the qualitative and quantitative features of diaspores in fur. We recorded 25,650 diaspores of 141 plant taxa (with 107 taxa and 23,350 diaspores determined to species level), using three different methods: (i) the “whole-coat method”, (ii) the “part-of-thecoat method” and (iii) a “seedling emergence method”. A comparison of these techniques revealed that the “wholecoat method” provided the highest number of diaspores and plant species. All diaspores were clustered into five emergent groups based on seven functional traits (diaspore weight, length, width, height, volume, specific weight and the diaspore surface structure). Our research revealed that sheep represent an important dispersal vector, since about half of the plant species recorded in the pastures were found as diaspores in fur. This study contributes to knowledge about the modes of seed dispersal in seminatural grasslands. Taking into account that livestock play a key role in vegetation dynamics, understanding their effects on seed dispersal is essential for conservation and restoration of these species-rich grassland communities.

Drought, cadmium (Cd) stress, and root lesion nematode (RLN) infection are three of the most important stresses affecting ramie growth and development; therefore, ramie breeding programs focus on their management more than on any other abiotic or biotic stresses. The fact that only a small number of stress-responsive transcription factors (TFs) have been identified so far is a major obstacle in the elucidation of mechanisms regulating the response to these three stresses in ramie. In this study, in order to uncover more stress-responsive TFs, a total of 179 nonredundant genes with full-length open reading frames from the MYB, AP2/ERF, bZIP, HD-ZIP, and COL families were obtained by searching for against the ramie transcriptome. Expression pattern analysis demonstrated that most of these genes showed relatively higher expression in the stem xylem and bast than in other tissues. Among these genes, 96 genes were found to be involved in responses to drought, Cd exposure, or RLN-infection. The expression of 54 of these genes was regulated by at least two stresses. These stress-responsive TFs probably have roles in the regulation of stress tolerance. The discovery of these stress-responsive TFs will be helpful for furthering our understanding of the mechanisms that regulate stress responses in ramie.

Production of IL-2 by CD4 + T cells is shown to be suppressed in pregnant women infected by bacteria such as Salmonella typhi. Elephantopus scaber and Sauropus androgynus may be used as herbal supplement to ameliorate this condition. This study aimed to investigate the efficacy of E. scaber and S. androgynus formulation to promote lymphoid proliferation and CD4 cytokine productions. The pregnant mice were randomly divided into seven experimental groups: K- (control), K+ (with S. typhi ), P1 ( S. typhi + E. scaber 100%), P2 ( S. typhi E. scaber 75% and S. androgynus 25%), P3 ( S. typhi + E. scaber 50% and S. androgynus 50%), P4 ( S. typhi + scaber 25% and S. androgynus 75%), and P5 ( S. typhi S. androgynus 100%). FACS analysis was done on day 18. Typhoid fever caused decreasing IL-2 and IFN-γ and in contrast increasing IL-4 production. In this experiment we clearly showed that typhoid fever decreased the amount of CD4 T cells but rather increased the amount of B cells. Formulation of E. scaber and S. andogynus was able to ameliorate the condition by increasing IL-2, IFN-γ, CD4 T cells and decreasing both IL-4 cytokine production and the amount of B cells.

In this study, we aimed to evaluate whether exposure to caffeine in the early stages of development affect AdoR mRNA expression levels in the fruit fly ( Drosophila melanogaster ) and how this will relate to the developmental success of flies. Adenosine receptors are seen as the most important biochemical targets of caffeine. Simultaneously adenosine signaling orchestrates the development and growth of insects. We demonstrate that AdoR mRNA expression in D. melanogaster is persistent from early stages till imago. Strong alterations in AdoR expression were observed in larvae that had been treated with caffeine. However, after the imaginal molt, the differences in AdoR expression between the insects from all of the test groups evened out despite a wide range of developmental success in the groups. Taken together, these results suggest that caffeine affects the expression of its cellular targets even from the early stages of fruit fly development and thus there is a significantly lower larvae-to-adult survival rate. Moreover, we also proved that the expression of AdoR undergoes a peculiar reset during the maturation of D. melanogaster despite the conditions in which larvae developed.

The purpose of this study was to evaluate the beneficial effects of magnesium (Mg) supplementation upon carbon tetrachloride (CCl 4 ) toxicity. Our study was carried out on 24 Wistar male rats divided into 4 batches. During a 6 week period, one group served as a control, two groups received Mg (after 4 weeks one of these groups was then treated with CCl 4 ), and a final group was treated with CCl 4 only. Under our experimental conditions, CCl 4 poisoning resulted in oxidative stress indicated by a significant increase in lipid peroxidation level in renal tissues. The blood levels of creatinine and urea increased while the blood level of uric acid and proteins decreased. CCl 4 also induced an increase in superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity in kidneys, in the number of red blood cells (RBC), and in hemoglobin content (Hb) and mean cell hemoglobin concentration (MCHC). However, white blood cell count (WBC), platelet count (Pl) and catalase activity (CAT) all decreased significantly. Treatment with Mg was found to alleviate most of CCl 4 -induced damage by decreasing lipid peroxidation and by correcting changed hematological parameters, and catalase, glutathione peroxidase and superoxide-dismutase activities. The results provide strong evidence that Mg supplementation is beneficial in protecting the kidneys from CCl 4 toxicity.

Botrytis spp. cause several diseases in Allium crops and depending on meteorological conditions economic losses can exceed 50%. Forecasting models improve plant protection and sometimes reduce consumption of fungicides, because applications are made precisely during the favourable periods for disease development. Our aim was to evaluate the iMETOS®sm B. cinerea forecasting model as an effective onion leaf fleck management system and estimate the variability of onion bulb pathogens during storage. Assessment of forecasting model data showed that favourable conditions for leaf fleck development arise in July, but greatly depend on that year’s meteorological conditions. During an experimental year the first sprayings with fungicides were applied as forecasted from the model, which resulted in application 19, 6 and 23 days earlier than conventional treatment application times. In 2012-2014 iMETOS®sm treatment yield increased by 3.51 t ha -1 , 3.87 t ha -1 and3.40 t ha -1 relative to the control. During storage most frequent injuries were fungal (44%) and bacterial (41%), followed by insects (7%) and physiological (9%). The highest prevalence of injuries was detected after 2 months of storage.

An air diffusion based system (Airx) was developed to control the dissolved oxygen levels in aquaculture sea cages. The system was introduced and then tested for 37 days in a sea bass sea cage (aerated cage). A second sea bass sea cage, without the AirX, was used as a control. Oxygen levels were measured in both cages at the start of the trial, before the AirX system was introduced, and during the working period of the AirX system. Fish samples were collected 15 days after the AirX system was introduced and at the end of the experiment. Blood smears were prepared and examined microscopically. Erythrocyte major axis, minor axis and area of fish erythrocytes were measured. Leucocyte differentiation was also examined. In the control cage, the fish had significantly larger red blood cells when compared with the red blood cells of the fish in the aerated cage. Histological examination of the gills and brain revealed no morphological differences or alterations between the two groups of fish. This study demonstrated that an air diffuser system could improve the water quality of fish farmed in sea cages and enhance sea bass physiological performance, especially if DO levels fall below 60% oxygen saturation.

This experiment examined the influence of two different silage additives of biological ( Lactococcus lactis, Lactobacillus plantarum, Enterococcus faecium , enzyme xylanase) and chemical (43% formic acid, 30% ammonium formate, 10% propionic acid, 2% benzoic acid) types on biogenic amines concentration, nutrient content, fermentation process, and microbiologic indicators in lucerne ( Medicago sativa ) silage after 90 days of fermentation. The biological additive significantly (P < 0.05) increased putrescine (+51%), lactic acid (+11%) and protein content (+11%) in comparison with control silage. It significantly decreased cadaverine (−29%), histamine (−57%), spermidine (−15%), spermine (−55%), acetic acid (−40%), ethanol (−55%), ammonium (−25%) and ash (−9%). After the chemical-additive treatment, greater amounts of histamine and tyramine were recorded. Significant decrease was observed in the concentrations of putrescine (−18%), cadaverine (−55%), spermidine (−47%), spermine (−45%), lactic acid (−16%), acetic acid (−46%), ammonium (−59%), ash (−13%) and fat (−24%). Populations of bacteria associated with lactic acid fermentation, moulds, yeasts, enterobacteria and total microorganisms count were also influenced. Both biological and chemical additives can be highly recommended for producing high-quality silages meeting hygienic requirements. In lucerne silage, the chemical preservative showed a stronger effect in achieving the health safety of silage compared to the biological inoculant.

The emergence of antibiotic resistant bacterial pathogens is becoming a major challenge for patient care. The utilization of alternative therapies for infectious diseases other than antibiotics is an urgent need of today medical practice. The utilization of lytic bacteriophages and their gene products as therapeutic agents against antibiotic resistant bacteria is one of the convincing alternative approaches. Here we present the isolation and characterization of three lytic bacteriophages TSE1-3 against Enterobacter cloacae from sewage effluent. The isolates maintained antibacterial activity for 10 hours of incubation followed by the development of phage resistance. Their stability at different temperatures and pH, established their possible application in phage therapy. The highest activity of the phages was observed at 37°C and pH 7.0, while they gave lytic activity up to 60°C. The latent period of all the TSE phages was 20 minutes, while the burst size was 360 for TSE1, 270 for TSE2 and 311 for TSE3. The phages were harboring double-stranded DNA larger than 12kb in size. Further research into the phages genome and proteins, animal experiments, delivery parameters and clinical trials may lead to their utilization in phage therapy.

Background : We studied in vitro and vivo antioxidant and prooxidant abilities of aqueous extracts from Rosa canina L., Rhodiola rosea L., Hypericum perforatum L., and Gentiana lutea L. Methodology : Total antioxidant capacity was measured by four assays (phosphomolybdate method, Fe 3 +-reducing activity, ABTS •+ scavenging, H 2 O 2 scavenging). Prooxidant activity was estimated by H 2 O 2 production. Yeast viability in the presence of H 2 O 2 and/or plant extracts was determined by plating or by counting live cells’ number. Results : Plant extracts differed in the total phenolic content ( R. canina > R. rosea > H. perforatum > G. lutea) which clearly correlated with their ABTS •+ scavenging activity (R 2 = 0.963). H 2 O 2 scavenging activity was not clearly associated with plant phenol levels and was significantly higher in acidic, than in alkaline medium. In line with this, plant extracts effectively protected yeast S. cereviasiae against H 2 O 2 and stimulated reproductive ability of yeast cells at acidic but not at alkaline pH. At alkaline pH, plant extracts produced certain amounts of H 2 O 2 which were related to their phenolic content. Conclusion : The antioxidant activity of plant extracts is decreased at alkaline pH with an increase in the prooxidant activity. It reduces protective capacity of plant extracts against oxidative and other stresses in vivo .

Background Running economy (RE), expresses the relationship between the energy cost of running (Cr) and the work performed by a runner and is an predictor of performance. Given the intense effort of marathon runners during training and competition and the dearth of studies that address performance and cytokines in this population, the objective of the current study was to investigate the relationship between RE and cytokines in marathon runners. Methods A total of 22 recreational marathon runners were examined. Using data obtained from VO 2max assessments and sub-maximal tests, the following formula was applied to determine RE: Cr (mLO 2 ·kg -1 ·km -1 ) = VO 2 (mL·kg -1 ·h -1 ) × 60 ÷ speed (km·h -1 ). Results Cr values shows no correlation with levels of the serum IL-1β, IL-4, IL-8, IL-10 and TNF-a 24h before, immediately after or 72h after the completion of an official marathon. However, the IL-6 level shows a significant correlation with Cr. Discussion and conclusion The relationship between higher values of IL-6 and lower RE leads to the hypothesis of a physical under-recovery state by some athletes. Considering the stress caused by training, associated with the higher energetic cost in less economic athletes, it’s possible that the period of resting may not totally compensate for the inflammatory state.

Chromatin remodeling in DNA is fundamental to gene expression, DNA replication and repair processes. Methylation of promoter regions of tumor-suppressor genes and histone deacetylation leads to gene silencing and transcriptionally repressive chromatin. For the past few decades DNA methylation agents became very attractive as the targets for cancer therapy. The purpose of this work was to examine the effects of DNMT inhibitor procaine on growth inhibition, apoptosis and differentiation of human leukaemia cells. The changes in expression of genes, proteins and histone modifications caused by procaine were evaluated under different treatments. We demonstrated that procaine arrests growth of human leukaemia cells and in combination with all-trans retinoic acid (ATRA) induces cancer cell differentiation. Procaine causes reduction of expression of DNA methyltransferases as well. The treatment of human leukaemia cells with procaine increase the expression of molecules associated with differentiation (CD11b, E-cadherin, G-CSF) and apoptosis (PPARγ). Moreover, the examined DNMT inhibitor enhances certain gene transcription activation via chromatin remodelling – the changes in histone H3K4(Me)3 and H3K9Ac/S10P modifications were detected. Our results suggest, that DNMT inhibitor – procaine, can be used for further investigations on epigenetic differentiation therapy of leukaemia cells especially when used in combination with retinoic acid.

Although almost all biological processes are mediated by a variety of proteins, it is important to bring to spotlight recent experimental and clinical research advances that had their focus on highlighting and taking advantage of the roles of several strategic proteins in order to gain more understanding of cancer biology. Proteins have a major stake in the initiation, progression, sustenance and completion of cellular processes, and have also demonstrated their vital roles in cancer processes. The characteristic functions of proteins and modified proteins have been utilized in the understanding and treatment of cancer. Recent insights in such roles and applications include linker histone H1.2 in the compaction of chromatin and gene silencing via the recognition of H3K27me3; c-Jun with Fra-2/c-Fos in the promotion of aggressive tumour phenotypes in tongue cancer; the use of sodium channelinhibiting agents targeting the transmembrane protein in breast, colon and prostate cancer; SET-mediated activities; protein interaction networks in glioma; Gpnmb significance as a biomarker; β-carbolines inhibition on Wnt/β-catenin signaling; p53 mutants co-opt chromatin pathways; Bone morphogenetic protein 4 as regulator of the behaviors of cancer cell; Brain-Expressed X-linked (BEX) proteins in human cancers; targeting CDK4/6 including protein kinases to make a reversal of multidrug resistance in sarcoma. In-depth knowledge of Proteomics will go a long way in helping us uncover a lot more strategies that will help us in the long fight against cancer.

Cultivation of the Burgundy truffle, Tuber aestivum Vittad., has become a new agricultural alternative in Poland. For rural economies, the concept of landscaping is often considerably more beneficial than conventional agriculture and promotes reforestation, as well as land-use stability. Considering examples from France, Italy, Hungary and Spain, truffle cultivation stimulates economic and social development of small, rural communities. Because there is no long tradition of truffle orchards in Poland, knowledge regarding the environmental factors regulating the formation of fruiting bodies of T. aestivum is limited. Thus, knowledge concerning ectomycorrhizal communities of T. aestivum host species is crucial to ensuring successful Burgundy truffle production. We investigated the persistence of T. aestivum ectomycorrhizae on roots of hazel ( Corylus avellana L.) and oak ( Quercus robur L.) and checked the host-species influence on community structure of ectomycorrhizal fungi. The study was conducted in an experimental plantation located in eastern Poland and established in 2008. We demonstrated that the number of fungal taxa was not significantly different between oak and hazel. However, the species composition differed between these two host trees. During the three-year study, we observed that species richness did not increase with the age of the plantation.

Human leukocyte antigen G (HLA-G) is a non-classical HLA class I protein with various immunosuppressive functions. Besides its profound effect to induce fetal tolerance, HLA-G has been also found to enhance graft acceptance. The aim of the study was to analyse the association between HLA-G 14 bp insertion/deletion polymorphism, soluble HLA-G level and kidney graft outcome in the Slovak population. We investigated 69 kidney transplant recipients aged 27–65 years. Out of this group, 37 recipients developed acute rejection, confirmed by biopsy, and 32 patients had stable allograft function. Plasma was obtained from recipients at 1 day before transplantation and analyzed by ELISA. Genotyping of HLA-G polymorphism was performed by PCR. Significantly higher pre-transplantation levels of sHLA-G were found in the group with stable allograft function in comparison to group with acute rejection (P = 0.0409). In the homozygous −14/−14 recipients with stable allograft function, significantly higher values of sHLA-G were determined in comparison to the recipients with acute rejection (P = 0.0052). The study revealed an association between 14 bp deletion polymorphism and soluble HLA-G level that is proportional to kidney graft acceptance. It is suggested that pre-transplantation levels of soluble HLA-G should be monitored as additional marker to predict kidney graft outcome.

As a novel biomarker from the STEAP family, STEAP2 encodes six transmembrane epithelial antigens to prostate cancer. The overexpression of STEAP2 is predicted as the second most common cancer in the world that is responsible for male cancer-related deaths. Nonsynonymous SNPs are important group of SNPs which lead to alternations in encoded polypeptides. Changes in the amino acid sequence of gene products can lead to abnormal tissue function. The present study firstly sorted out those SNPs which exist in the coding region of STEAP2 and evaluated their impact through computational tools. Secondly, the three-dimensional structure of STEAP2 was formed through I-TASSER and validated by different software. Genomic data has been retrieved from the 1000 Genome project and Ensembl and subsequently analysed using computational tools. Out of 177 non-synonymous single nucleotide polymorphisms (nsSNPs) within the coding region, 42 mis-sense SNPs have been predicted as deleterious by all analyses. Our research shows a welldesigned computational methodology to inspect the prostate cancer associated nsSNPs. It can be concluded that these nsSNPs can play their role in the up-regulation of STEAP2 which further leads to progression of prostate cancer. It can benefit scientists in the handling of cancerassociated diseases related to STEAP2 through developing novel drug therapies.

Objective This study was to investigate the single nucleotide polymorphism (SNP) in the interferon regulatory factor 6 (IRF6) gene in healthy residents of Guangdong Province, China, for further analysis of their associations with the development of cleft lip with or without palate (CL/P). Methodology DNA was extracted from blood samples of 13 healthy residents. IRF6 genes were sequenced and analyzed by alignment to the reference sequences in GenBank. Results The IRF6 genes containing 45.21% GC were 25016–25046 bp in length, and had 2215-bp exons and a 1404-bp coding region. There were 65 SNPs, including 58 SNPs in the 5′-untranslated region or introns and 7 SNPs in the exons. These sites had two alleles, including 39 transition sites and 26 transversion sites. Five novel SNPs were identified. c. 459G>T and c. 820G>A in the coding region represented silent and Val274Ile mutations, respectively. The SNPs made two sequences (GenBank HQ875393 and JF346417). Conclusions The sequences and SNP profiles of the IRF6 gene in healthy residents of Guangdong Province are consistent with the one in GenBank but with slight variations. Our findings form a basis for further investigation of the association between IRF6 gene polymorphisms and CL/P in residents of Guangdong Province.

This paper deals with the assessment of cultivation of bread wheat ( Triticum aestivum L.) and oat ( Avena sativa ) grown in Central Europe within the conventional and organic farming systems in terms of greenhouse gas emissions and economic profitability. Organic farming may be one of the tools for mitigation of greenhouse gas emissions from agricultural production. In the context of crop production, cereals rank among the most commonly grown crops and therefore bread wheat and oat were chosen. The Climate change impact category was assessed within the simplified LCA method and the production of greenhouse gas emissions expressed in CO 2 e per the production unit was calculated. Economic balance of the cultivation of monitored cereals was compiled based on the yields, farm gate prices and costs. On its basis, the cultivation of wheat within the organic farming system appears to be the most profitable. From an environmental point of view, the emission load of the organic farming system is reduced by 8.04 % within the wheat production and by 15.46 % within the oat cultivation. Therefore, the organic farming system in the Czech Republic appears to be more environmentally friendly and economically efficient within the cereals production.

Central-eastern Europe is an endemic region for cystic echinococcosis where multiple species of intermediate hosts are commonly infected with Echinococcus granulosus sensu lato tapeworms of major medical and veterinary importance. Investigations of the genetic variation of 25 Echinococcus isolates from five countries (Slovakia, Romania, Ukraine, Hungary, Poland) were undertaken using three mitochondrial DNA markers. The 18 isolates from pigs derived from Slovakia and Ukraine and the four human isolates from Slovakia, Poland and Ukraine were identified as E. canadensis G 7, whereas the three human isolates from Romania and Hungary were classified as E. granulosus sensu stricto G1. This study reports the first confirmed human case of E. granulosus s.s. in Hungary. The haplotype G7A with two polymorphic sites relative to the most common regional variant of E. canadensis G7 was recorded in both pigs from Ukraine and in a single pig isolate from Slovakia. The results of this study support the circumstantial evidence that E. canadensis G7 with low infectivity for humans is highly prevalent in the northern parts of the region (Poland, Slovakia, forest-steppe zone of Ukraine), while infections with E. granulosus s.s. which are highly infectious for humans are more commonly encountered in Romania and Hungary.

Non-Hodgkin lymphoma (NHL) is a serious disease, with a high proportion of mortality. Molecular genetic abnormalities are very common in NHL, but specific characterization in accordance to molecular genetics for lymphoma subtypes is not yet completed. This article summarizes the relationship between B- and T-NHL and molecular genetics. We focus on NHL subtypes and emphasize its features to figure out what is exposed about NHL genetics. The basis of this method is collection of biological specimens for genomic and genetic analyses. This summary may help to prompt prediction of outcomes and guide therapy in the future.

Long noncoding RNAs (lncRNAs) are nonprotein coding transcripts longer than 200 nucleotides. Aberrant expression of lncRNAs has been found to be associated with hepatocellular carcinoma, one of the most malignant tumors. In this paper, we give a systematic and comprehensive review of existing literature about the involvement of lncRNAs in hepatocellular carcinoma. To date, evidence suggests that a number of lncRNAs, including HEIH, H19, HOTAIR, MALAT1, and PVT1, may regulate the transcription of target genes by recruiting histone-modifying complexes. Under certain circumstances, lncRNAs form RNA-dsDNA triplexes. Certain lncRNAs, such as HULC, HOTAIR, H19, HOTTIP and PTENP1, exhibit their biological roles by associating with microRNAs (miRNAs). In addition, by complementary base pairing with mRNAs or forming complexes with RNA binding proteins (RBPs), lncRNA-ATB, MALAT1 and PCNA-AS1 may mediate mRNA stability and splicing. In conclusion, interactions with DNA, RNA and proteins appears to be involved in lncRNAs’ participation in tumorigenesis and developmental processes related to hepatocellular carcinoma.

Wnt/β-catenin signaling has been proved to play an important role in the development and promotion of cancer metastasis. The activation of Wnt signals can lead to duplicating, updating, metastasizing and relapsing. The Wnt signaling pathway is mainly divided into the Wnt/β-catenin pathway and the Wnt/calcium pathway. A better understanding of all the diverse functions of Wnt and their molecular mechanisms has evoked prevailing interest in identifying additional targets related to the Wnt /β-catenin pathways in breast cancer. A number of new target, related to Wnt /β-catenin pathways have been identified in recent years, including NOP14, BKCa channels, Emilin2, WISP, MicroRNAs, NRBP1, TRAF4, and Wntless. In this review, we will introduce the new targets related to the Wnt /β-catenin pathways in breast cancer.

Objective It is already known that long non-coding RNA growth arrest-specific 5 (GAS5) is downregulated in human colorectal cancer (CRC) cells inhibiting cell proliferation. We further analyzed its involvement in cell cycle distribution and apoptosis induction. Methods We measured the expression level of GAS5 in CRC tissues and cell lines with the corresponding non-tumoral cells. We also analyzed the roles of GAS5 in modulation of cell growth, cell cycle distribution and apoptosis by the CCK-8 method and flow cytometry. Western blots were performed to evaluate the protein level of cyclin D1 and p21 after overexpression of GAS5 Results GAS5 expression was significantly reduced in CRC samples and cell lines. Overexpression of GAS5 induced cell growth arrest and induced cell apoptosis in vitro. Meanwhile, we found that the growth suppressive role of GAS5 might be attributed to the inhibition of G1-S phase transition, reflected by the downregulation of cyclin D1 and upregulation of p21. Conclusion Our results demonstrate that GAS5 is a crucial tumor suppressor in human CRC cells.

Breast cancer is the most prevalent cancer in women worldwide. Numerous studies have suggested that the E-cadherin adhesion system is dysregulated in cancer cells. These impaired functions of E-cadherin contribute to releasing cancer cells from the primary lesion to cell dedifferentiation. Some studies have shown that polymorphism may affect E-cadherin expression, and then play a role in susceptibility to breast cancer. However, the results remained controversial. In this short review, we summarize the functions of E-cadherin and the signaling pathways it regulates, and assess the association between CDH1 polymorphisms and breast cancer susceptibility. This study suggests that genetic variation in CDH1 and -160C/A polymorphism may have an association with breast cancer risk. The assessment of CDH1 polymorphisms may be used for the identification of patients suitable for anti- CDH1 therapy.

Brain tumors include primary tumors of various intracranial tissue and secondary intracranial tumors that transferred from other parts of the body. Secondary intracranial tumors are especially prevalent in patients with lung cancer. The mechanisms of lung cancer with brain metastases are complicated, they are affected by a variety of factors. Thus, identifying the mechanisms of lung cancer with brain metastases will have far-reaching meanings both for clinic pharmacy research and for a better quality of life for patients; Brain metastases from lung cancer represent a prevalent and challenging clinical dilemma, and some research suggests that the outcomes and characteristics of brain metastases that result from lung cancer primary sites are perhaps different than those from other primary sites, therefore increasing the difficulty of clinical treatment. Despite steady research developments during recent years, the survival rates remain poor. The mechanisms and therapeutic options for treating brain metastases arising from lung cancer are review in this article.

Renal cell carcinoma (RCC) is the most frequent form of renal cancer, and is associated with a high frequency of metastasis. While, there is few therapeutic methods can substantially prolong survival. Superior to cytokine therapy with IL-2 and/or IFN-a, several newer targeted treatments are available for the treatment of patients with advanced conventional (clear cell) renal cell carcinoma (RCC), which received improved outcomes. These newer targeted treatments include the multi-targeted tyrosine kinase inhibitors (TKIs, sorafenib, sunitinib, pazopanib, and axitinib), the humanised antivascular endothelial growth factor (VEGF) monoclonal antibody [bevacizumab combined with interferon (IFN)-a], and mTOR (mammalian target of rapamycin) complex 1 kinase inhibitors (everolimus and temsirolimus). However, these targeted drugs are still associated with limited efficacy and high toxicity, so there is still a strong need for further discovery of new targeted drugs. In the present manuscript, we summarize newly-presented potential targeted drugs for RCC, classified by drug characteristic, small molecule, small molecule combination, monoclonal antibody, polysaccharides, organometals and peptides.

Objectives Colorectal cancer (CRC) is among the most common types of malignancies in the worldwide, and microRNAs (miRNAs) emerge as key regulators in carcinogenesis and tumor progression. Here we intended to address the expression and function of miR-132 in CRC cells. Methods Paired CRC tissues and several established cell lines were firstly collected. We performed qPCR to detect the expression of miR-132 in these tissues and cell lines. Cell proliferation and apoptosis were respectively monitored by CCK-8 assay and Annexin-V/PI staining followed by flow cytometry, after miR-132 was transiently overexpressed in RKO cells. Afterwards, Luciferase reporter assays were performed to examine the targeting of YAP1 by miR-132. Finally, qPCR and western blotting were also carried out to validate this targeting. Results MiR-132 was significantly decreased in CRC and its overexpression in RKO cells exerted tumor suppressing effects, including cell growth arrest and apoptosis promotion. Additionally, we proved that miR-132 could negatively regulate the expression of YAP1. Conclusion Our findings suggested that miR-132 was downregulated in CRC, and played as a tumor suppressor to inhibit cell proliferation and induce apoptosis. And these anti-tumor activities might be related with the targeting of YAP1 by miR-132.

Retinoblastoma protein-interacting zinc finger gene RIZ encodes two different protein products, RIZ1 and RIZ2. Observations suggest that RIZ1 is a tumor suppressor, while RIZ2 acts as a negative regulator of RIZ1 and may play a positive role in oncogenesis. The imbalance amount of RIZ1 and RIZ2 may be involved in cancer development. In this study we detected the expression levels of RIZ1 and RIZ2 mRNA in human esophageal squamous cell carcinoma (ESCC) tissue specimens, reexpressed RIZ1 in the human ESCC cell line EC109 in which RIZ1 mRNA level was not detected, examined the changes of RIZ1 and RIZ2 mRNA expression, investigated the changes of proliferation, and apoptosis of the cells. We found that RIZ1 mRNA expression is commonly decreased or at undetectable level in human esophageal squamous cancer tissue specimens compared to the normal tissue specimens, while RIZ2 is usually expressed. With the forced expression of RIZ1, RIZ2 mRNA expression did not change, The ESCC cell proliferation was inhibited and apoptosis was induced. This unusual yinyang fashion of RIZ1/RIZ2 may contribute to the progression of ESCC.

Vulvar carcinoma is a rare tumor occurring in female patients. Though more than 40% of vulva cancers are due to the infection of human papillomavirus (HPV), understanding of HPV and vulvar carcinoma is insufficient. HPV expression is regulated by cellular and viral transcription factors that bind to specific elements within the ligase chain reaction. These proteins bind with different affinity to host cell proteins and disrupt normal epithelial differentiation and apoptosis. Immunotherapy does not target tumors, but instead targets the host immune system. Active immunotherapy is tumor-targeting or immune-targeting monoclonal antibodies and vaccines. Nonspecific active immunotherapy is mainly cytokine therapy. In the treatment and prevention of HPV, the most popular research projects were regarding peptide, recombinant protein and DNA-based vaccines, recombinant virus and other targets in HPV infection. Since the cervix and vulva are both susceptible areas, these studies may be able to help reduce prevalence of vulvar precancerous lesions and prevent all cancers caused by HPV.

Objectives The long non-coding RNA (lncRNA) IRAIN has been verified to have key roles in tumor biology. The aim of this study was to explore its expression and biological functions in human renal cell carcinoma (RCC) cells. Methods Quantitative RT-PCR was applied to detect the RNA expression of IRAIN in RCC tissues and cell lines when compared with respective controls. MTT and flow cytometry methods were respectively used to monitor the cell proliferation and apoptosis of 786-O cells after IRAIN was overexpressed. Altered expression of cyclin D1 and Bax was determined by immunoblotting. Xenograft models were finally carried out to confirm the roles of IRAIN in RCC in vivo . Results IRAIN expression was found to be remarkably decreased in RCC tissues and cell lines. Its overexpression in 786-O cells significantly inhibited cell proliferation and promoted apoptosis. We further demonstrated that cyclin D1 was reduced while apoptosis promoting protein Bax was elevated in IRAIN-overexpressed 786-O cells. Importantly, we found that IRAIN overexpression could suppress in vivo tumorigenesis of RCC, reflected by tumor volume and tumor weight measurement. Conclusion IRAIN might serve as a novel tumor suppressing lncRNA and a potential therapeutic target in RCC treatment.

Objective Recently, the role of long noncoding RNAs (lncRNAs) in human colorectal cancer (CRC) has been a subject of intense focus. We set out to determine the function of one lncRNA, termed urothelial carcinoma-associated 1 (UCA1) in CRC cell proliferation and its underlying mechanisms. Methods Quantitative real-time PCR (qRT-PCR) was applied to detect the expression level of UCA1 in 50 pairs of CRC samples compared with non-tumor colon tissues. Cell growth was determined using the Cell Counting Kit-8 (CCK-8). Western blotting was carried out to analyze the related protein expression. Flow cytometry was done to evaluate cell apoptosis by UCA1 inhibition. Results We found an increased expression of UCA1 in CRC samples. Knockdown of UCA1 in HCT116 cells induced a decrease in cell proliferation rate compared to control samples. This oncogenic activity may be enhanced through p53/ p21 signaling. Conclusion Our results supported the hypothesis that upregulation of UCA1 contributes to the unlimited proliferation rate of CRC cells, at least partially through the negative regulation of p53/p21 signaling pathway. Finally, we found that UCA1 merely influenced CRC cell apoptosis.

Objectives : As the member of the Fox family of transcription factors, Forkhead box M1 (FoxM1) is known to be critical in pathogenesis and development of many solid tumors. However, the clinical value and expression pattern in non-small cell lung cancer (NSCLC) is still poorly understood. Methods : In this study, real-time PCR was mainly applied to examine the gene expression levels of FoxM1 in 120 pairs of clinical NSCLC tissues, which were classified into different groups according to smoking status, lymph node metastasis, and tumor grade. By utilizing the online Kaplan-Meier plotter, overall survival analysis was performed to study the correlation between FoxM1 expression and prognosis of lung cancer (LC) patients. Afterwards, the correlation of FoxM1 gene expression and the clinical pathological parameters was examined by κ 2 test in these 120 NSCLC patients. Results : FoxM1 was found to be aberrantly upregulated in NSCLC patients, and its overexpression was correlated to groups designated as smokers, cases of positive lymph node metastasis and cases of advanced tumor grades. Online survival analysis showed that high expression of FoxM1 predicted shorter overall survival of NSCLC patients. Additionally, FoxM1 upregulation was statistically correlated with positive smoking history, lymph node metastasis and higher tumor grades. Conclusion : FoxM1 is overexpressed in cancerous tissues and is associated with the poor prognosis of NSCLC patients. Our results provide insights into the utility of FoxM1 as an important biomarker and prognostic factor for NSCLC.

The oncogenic driver mutations have been found that not only have potential sensitivity to epidermal growth factor receptor but also can inhibit anaplastic lymphoma kinase tyrosine kinase; more and more interest has been evoked in discovering additional targets to non-small cell lung cancer (NSCLC). Recently, many novel underlying oncogenic gene alterations have been identified, such as HER2 insertions, BRAF mutations, PIK3 mutations, FGFR1 amplifications, DDR2 mutations, KRAS mutations, MET amplification, ROS1 rearrangements, ALK rearrangements, and RET rearrangements. In this review, we will discuss the discovery of these potential targets and the application of each in NSCLC and of small molecular inhibitors on these potential targets.

Objectives Lung cancer is the leading cause of cancer-related death, with non-small cell lung cancer (NSCLC) accounting for more than 80% of all lung cancer cases. The aim of this study was to investigate the function and underlying mechanism of microRNA-192 (miR-192) in metastasis of NSCLC cells. Methods Real-time PCR was applied to quantify the expression of miR-192 in NSCLC tissues and cell lines, matched with their corresponding controls. The biological roles of miR-192 were studied in NSCLC cells using the wound healing and trans well invasion assays. Real-time PCR and western blot were used to evaluate the regulation of ZEB2 by miR- 192. Results MiR-192 was expressed significantly lower in NSCLC tissues/cells when compared with controls. Ectopic expression of miR-192 strongly inhibited cell migration and invasion in NSCLC A549 cells. Further investigation revealed ZEB2, an EMT regulator, was one of the downstream targets regulated by miR-192. Conclusion These results suggested that miR-192 inhibits the metastasis of NSCLC cells by targeting ZEB2, and thus is an important tumor suppressor.

Objective Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is already known to be involved in the development and progression of many types of tumors. In the present study, we set to seek the role of MALAT1and the molecular mechanisms in breast cancer. Methods MALAT1 mRNA expression level was measured by real-time PCR in selected tissues and breast cancer cell lines. SiRNAs targeting MALAT1 were employed to knockdown the endogenous MALAT1. Then cell counting method and colony formation method were applied to reveal the proliferation changes after MALAT1 was suppressed. Afterwards, the mRNA and protein expression of growth related gene cyclinD1 (CCND1) were detected by RT-PCR and western blotting, respectively. Results We found a downregulation of MALAT1 expression in breast cancer cell lines and tissues. Inhibition of its expression led to enhanced cell proliferation and colony formation. Importantly, the mRNA and protein expression of CCND1 was significantly increased in MALAT1-depleted cells. Conclusion MALAT1 is a potential tumor suppressive long non-coding RNA that negatively regulates cell proliferation in breast cancer progression, via suppressing CCND1 expression.

Metastasis is an important reason for death of cancer patients which characterized as the formation of secondary cancers at distant sites. Epithelial– mesenchymal transition (EMT) is a dynamic process that appear to facilitate tumor metastasis in various cancers by switching epithelial cells into mesenchymal properties. Although previous investigation suggested a key role of EMT transcriptional factors in suppression of E-cadherin, the association of these factors with other cellular regulators in cancer metastasis need to be fully elucidated. Post-translational modifications (PTMs), such as acetylation and phosphorylation, have emerged as an important mechanism to modulate biological behavior of substrate proteins. In this review, we summarized protein modification and subsequent function changes of Snail, Twist and ZEB, as well as their influence on tumor progression. Acetylation of EMT transcriptional factors usually cause nuclear localization and/or protein stabilization thus contribute to E-cadherin repression. Besides, Twist and ZEB were phosphorylated by diverse kinases to promote metastasis in many cancers, while Snail was negatively regulated by phosphorylation to degradation. Then, the potential of therapy for metastasis by targeting PTMs-involved regulation of EMT transcriptional factors were discussed.

Objective To explore the correlation between the enhancer of zeste homolog 2 (EZH2) expression and clinicopathological features in colorectal cancer patients. Methods A total of sixty-six patients with colorectal carcinoma were admitted to our general surgery department from January 2011 to December 2014. The EZH2 expression levels in the cancer tissues (CTs) from the 66 patients with colorectal cancer and those in distant normal colorectal tissues from 30 cases were examined through immunohistochemistry and western blotting assays. The relationship between the expression of EZH2 and the clinicopathological features and prognosis of the patients was analyzed. Results EZH2 in colorectal carcinoma tissues is granularly brown, predominantly expressed and diffused in the nuclei of tumor cells. Positive rates of EZH2 in intestinal CTs and in distant normal intestinal tissues are 62.12% (41/66) and 6.67% (2/30), respectively with significant difference (P < 0.05). Western blotting also confirmed its elevated expression in colorectal CTs. EZH2-positive expression in CTs was related to degree of differentiation, Duke staging, and tumor size (P < 0.05) but was unrelated to the patient’s gender, age or tumor site (P = 0.05). The 3-year progression-free survival (PFS) rates of the EZH2-positive group and the EZH2-negative group were 43.8% and 67.5%, respectively. The risk of disease progression of the EZH2-positive patients in the follow-up period was significantly higher than that of the EZH2-negative patients (HR = 2.49, 95% CI = 1.04–4.80, P < 0.05). Conclusion EZH2 is closely related to colorectal carcinoma development and disease progression, and thus could be used as a tumor biomarker that may indicate prognosis.

Objective The aim of this study was to investigate epidermal growth factor receptor (EGFR) expression in the gastric cancer tissues and the association of EGFR expression with clinical pathology characteristics. Methods We firstly analyzed the copy number alteration of EGFR gene in gastric cancer by utilizing the online TCGA resources. We then gathered 71 cases of gastric cancer patients from February 2010 to February 2015 in our hospital. The cancer tissues and normal gastric tissues were tested for EGFR expression by immunohistochemical (IHC) staining and western blotting methods. The association of EGFR expression with clinical pathology characteristics and the prognosis value of EGFR expression were evaluated by the statistical software. Results The copy number of EGFR gene was found to be increased in gastric cancer patients. Western blotting confirmed that EGFR protein was obviously upregulated in tested gastric cancer tissues. Additionally, our IHC results showed that the positive rates of EGFR expression in gastric carcinoid tumors (CTs) and distant normal gastric tissues were 45.1% (32/71) and 25.0% (9/36), respectively. The former was significantly higher than the latter (P < 0.05). The EGFR-positive expression in CTs was related to tumor size, invasion depth, and lymphatic metastasis. The median survival of the EGFR-positive patients was 15.6 months, which was less than that (23.0 months) of the EGFR-negative patients [HR = 2.12, 95%CI: 1.29-4.10 (P < 0.05)]. Also, online survival analysis showed that high expression of EGFR predicted shorter overall survival of gastric cancer patients. Conclusion EGFR was highly expressed in gastric cancer tissues and associated with poor prognosis.

Autophagy is a conserved catabolic process, which functions in maintenance of cellular homeostasis in eukaryotic cells. The self-eating process engulfs cellular long-lived proteins and organelles with double-membrane vesicles, and forms a so-called autophagosome. Degradation of contents via fusion with lysosome provides recycled building blocks for synthesis of new molecules during stress, e.g. starvation. Peiminine is a steroidal alkaloid extracted from Fritillaria thunbergii which is widely used in Traditional Chinese Medicine. Previously, peiminine has been identified to induce autophagy in human colorectal carcinoma cells. In this study, we further investigated whether peiminine could induce autophagic cell death via activating autophagy-related signaling pathway AMPK-mTOR-ULK by promoting SQSTM1(P62). Xenograft tumor growth in vivo suggested that both peiminine and starvation inhibit the growth of tumor size and weight, which was prominently enhanced when peiminine and starvation combined. The therapeutical effect of peiminine in cancer treatment is to be expected.

Objectives To investigate the expression as well as biological roles of activating transcription factor 3 (ATF3) in human hepatocellular carcinoma (HCC) cells. Methods Real time PCR and western blot were applied to detect the expression of ATF3 in human HCC specimens. MTT assay was used to analyze cell proliferation after ATF3 was overexpressed. The expression of cyclin D1 was detected by real time PCR in ATF3-silenced/-overexpressed cells and HCC samples. Correlation coefficient was finally analyzed between ATF3 and cyclin D1 in the HCC samples. Results The expression of ATF3 was found to be downregulated in tested HCC specimens. Cellular MTT assays showed that cell proliferation was suppressed in ATF3-overexpressing HepG2 cells. In addition, cyclin D1 gene expression exhibited negative correlation with ATF3 in both cell lines and tissue samples. Conclusion Low expression of ATF3 may function as a tumor suppressor via inhibiting cyclin D1 in HCC.

The EGFR signaling pathway plays an important role in the occurrence and development of many malignant tumors. It has become a hot spot in the treatment of advanced cancer. At present, the small molecule epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), has been shown to advanced non-small-cell lung cancer (NSCLC), has a marked drug resistance or has developed one. The EGFR signaling pathway regulates a variety of cellular functions, and its drug resistance may be related to a number of signal transduction pathways, including drug resistance mutations, structural activation, downstream signaling pathway activation and VEGF expression changes, and so on. In this paper, we review the production mechanism of EGFR-TKI drug resistance.

MicroRNA (miRNAs) is a group of small non-coding RNAs. It is involved in multiple cellular processes including proliferation, development, metabolism, differentiation and apoptosis; many of which are linked to several pathological conditions, including cancer. Lung cancer is one of the leading causes of mortality in the world: in 2008 for example, there were 163,000 deaths as a result of lung cancer. Despite technologies emerging which provide the potential for novel targeted therapies and improved early diagnoses, the overall rate of five-year survival still remains at only 15%. One reason for this disappointing statistic is related to the presentation of the disease, and specifically a lack of markers for early detection. Notably, the expression of some miRNAs has been reported to be involved in the diagnosis, classification and even prognosis of lung cancer. Tumorsuppressive and oncogenic miRNAs were found in lung carcinogenesis and the biological functions of these miRNAs have been validated in transplantable lung cancer models and human paired normal-malignant lung tissue banks. Some of these tumor-suppressive and oncogenic miRNAs related to lung cancer will be reviewed here. This article will focus on emphasing miRNAs effectiveness as a biomarker in lung carcinogenesis and candidate pharmacology. Furthermore, how these findings improve our understanding of lung cancer biology and therapy will also be discussed.

During 2012, about 782,500 new liver cancer cases were diagnosed and 745,500 deaths occurred all around the world. Liver cancer is the 2nd major cause of cancer death in men around the world and in underdeveloped countries. Dysregulation of the balance between proliferation and cell death, hepatitis B virus infection and hepatitis C virus chronic infection, and carcinogenic micro RNAs mainly contribute to the development and progression of liver cancer. Under physiological status, Src maintained the foundation of cells. While in liver cancer, it is known that the basic activities of cells are apparently disturbed possibly by Src. The mechanisms underlying the development and progression of liver cancer is needed to elucidate. In this study, we summarized newly found regulation pathway of SRC signaling, and clinical experience with inhibitors of Src signaling, such as, novel molecules that directly or indirectly targeted Src signaling which can be utilized in the treatment of liver cancer.

Cancer is the second leading cause of death worldwide. Traditional antitumor drugs exhibit severe cytotoxic and side effects. Lung cancer needs new and more effective treatment approaches. Coumarin derivatives can act on various tumor cells and show anti-proliferative activity through various mechanisms, including mitochondrial signaling cascades that regulate development and apoptosis of cells. Mitochondria-targeted coumarin derivatives have not been reported yet. Taking advantage of the fact that cancer cells frequently have higher mitochondria membrane potential, we synthesized a mitochondria-targeted 6-(nicotinamide) methyl coumarin by coupling 6-methyl coumarin to nicotinamide. Our results demonstrate that 6-(nicotinamide) methyl coumarin preferentially kills A549 cells through inducing A549 cells apoptosis, mediated by increasing ROS level and causing mitochondrial depolarization. Strikingly, the viability of the A31 cells treated with 6-(nicotinamide) methyl coumarin did not decrease, indicating that 6-(nicotinamide) methyl coumarin preferentially accumulates in A549 cells and A549 cells are much more susceptible to 6-(nicotinamide) methyl coumarin treatment compared with A31 cells.

Carboxymethyl chitosan grafted with ricinoleic acid (CMC-g-RA), an amphiphilic drug carrier, was synthetized, loaded with rotenone (Rot), and characterized for particles shape, zeta potential, loading efficiency and outdoor stability. Results show that as the ratio of carrier to drug increased, the formulation exhibited monodisperse nanoparticles negative surface charge. The loading efficiency of the formulation was up to 68%. The outdoor test also indicated that the formulation with the higher loading efficiency prevented Rot degradation in natural environments.

Population genetic diversity was estimated from forty-four individual ginseng ( Panax ginseng C.A. Meyer) plants collected from seven geographical populations located in Heilongjiang, Liaoning, and Jilin Provinces of China as well as the People’s Republic of Korea by using randomly amplified polymorphic DNA (RAPD) markers. Overall, 41 polymorphic loci were amplified using ten primer pairs. The polymorphism percentage ranged from 50% to 100% among seven local populations of ginseng, indicating that there is plentiful genetic diversity in wild ginseng populations. The genetic diversity at the species level was higher than that at the population level. Variance analysis showed that there was a significant difference among populations in genetic diversity. The genetic differentiation coefficient (i.e., F ST ) indicates that 43% of the variation occurred among populations, which indicates that substantial genetic differentiation occurred among populations. At the same time, the measured value of gene flow ( N m) was 0.66 based on the observed genetic differentiation coefficient among populations, suggesting there was moderate gene flow among populations.

Five different solvents (petroleum ether, chloroform, ethyl acetate, acetone, and distilled water) were used to extract antibacterial compounds from pineapple leaf fiber. Compounds extracted using acetone showed the greatest antibacterial effect against Escherichia coli , measured by inhibition zone diameter. Three extraction parameters including temperature, time and solid-liquid ratio were optimized through orthogonal experiment based on single factor investigations for achieving maximum active substance extraction rate and bacteriostatic effect. Results showed that using acetone, the optimum extraction conditions for temperature, time and solid-liquid ratio were 45°C, 8 h, and 1:40 (g/ml), respectively.

The main non-biodegradable compounds (soluble microbial product – SMP) of wastewater from the Maotai aromatic factories, located in the Chishui river region, were analyzed by UV spectroscopy, and by solid-phase extraction followed by gas chromatography coupled to mass spectrometry, after a two-stage biochemical treatment. The UV-Vis spectra revealed that the wastewater contained two double-bonds in conjugated systems (conjugated diene or α, β- unsaturated ketone, etc.) and simple non-conjugated chromophores containing n electrons from carbonyl groups or the like. The residual organic non-biodegradable substances were identified using SPE-GC/MS analysis as complex polymers containing hydroxyl, carbonyl, and carboxyl functional groups with multiple connections to either benzene rings or heterocyclic rings. As these compounds are difficult to remove by conventional biochemical treatments, our findings provide a scientific basis for the design of efficient new strategies to remove SMP from wastewater.

Pakistan occupies a significant global position in the growing of high quality cotton. The extensive application of pesticides on agricultural products leads to environmental risk due to toxic residues in air, water and soil. This study examined the chemodynamic effect of Deltamethrin on cotton fields. Samples were collected from the cotton fields of D.G. Khan, Pakistan and analyzed for heavy metal speciation patterns. Batch experiments were administered in order to study the adsorption of Deltamethrin in cotton fields. The effect of different factors including pH, adsorbate dose, and adsorbent mass on adsorption were studied. It was observed that in general, adsorption increased with increases in the mass of adsorbate, although the trends were irregular. Residual fractions of deltamethrin in the soil and water of cotton fields were analyzed to assess concentrations of xenobiotics bound to soil particles. Results indicated that such residues are significantly higher in soil samples due to high K oc in comparison to water, indicating the former is an efficient degradation agent. Results from the batch experiment resulted in 95% removal with alkaline pH and an adsorbent-adsorbate ratio of 250:1. These results may be used to environment friendly resource management policies.

This study was designed to find the best-fit probability distribution of annual maximum rainfall based on a twenty-four-hour sample in the northern regions of Pakistan using four probability distributions: normal, log-normal, log-Pearson type-III and Gumbel max. Based on the scores of goodness of fit tests, the normal distribution was found to be the best-fit probability distribution at the Mardan rainfall gauging station. The log-Pearson type-III distribution was found to be the best-fit probability distribution at the rest of the rainfall gauging stations. The maximum values of expected rainfall were calculated using the best-fit probability distributions and can be used by design engineers in future research.

Plant root foraging exhibits complex behaviors analogous to those of animals, including the adaptability to continuous changes in soil environments. In this work, we adapt the optimality principles in the study of plant root foraging behavior to create one possible bio-inspired optimization framework for solving complex engineering problems. This provides us with novel models of plant root foraging behavior and with new methods for global optimization. This framework is instantiated as a new search paradigm, which combines the root tip growth, branching, random walk, and death. We perform a comprehensive simulation to demonstrate that the proposed model accurately reflects the characteristics of natural plant root systems. In order to be able to climb the noise-filled gradients of nutrients in soil, the foraging behaviors of root systems are social and cooperative, and analogous to animal foraging behaviors.

The weevils Eucryptorrhynchus chinensis and Eucryptorrhynchus brandti (Coleoptera: Curculionidae), are two of the most important pests of the tree-of-heaven, Ailanthus altissima , which is found throughout China. In this study, the complete mitogenomes of the two weevils have been sequenced using Illumina HiSeq TM 2000. The mitogenomes of E. chinensis and E. brandt i are 15,628bp and 15,597bp long with A+T contents of 77.7% and 76.6%, respectively. Both species have typical circular mitochondrial genomes that encode 36 genes. Except the deficiency of tRNA-Ile, the gene composition and order of E. chinensis and E. brandti are identical to the inferred ancestral gene arrangement of insects. In both mitochondrial genomes, the start codons for COI and ND1 are AAT and TTG, respectively. A5bp motif (TACTA) is detected in intergenic region between the tRNA-Ser (UCN) and ND1 genes. The ATP8/ATP6 and ND4L/ND4 gene pairs appear to overlap four or seven nucleotides (ATAA/ATGATAA) in different reading frames. The complete sequences of AT-rich region have two regions including tandem repeats. The study identifies useful genetic markers for studying the population genetics, molecular identification and phylogeographics of Eucryptorrhynchus weevils. The features of the mitochondrial genomes are expected to be valuable in

This paper discusses the characteristics of source and sink for super hybrid rice and how to coordinate its source-sink relationships for high-yielding cultivation. It is known that super hybrid rice possesses a higher net photosynthetic rate than non-hybrid rice because of its higher grain-leaf area ratio, better stornata traits and less midday depression. However, the sink of super hybrid rice remains large due to its large and numerous spikelets. Furthermore, the relocation of assimilates is smooth in super hybrid rice because of its well-developed vascular structure. However, due to the very large sink of super hybrid rice, it is relatively inefficient in supplying of assimilate products to spikelets, in particular to inferior spikelets. Therefore, reducing the discrepancy between source and sink in super hybrid rice is essential for developing high-yielding cultivation. This can only be achieved by planting cultivars adapted for local environments, raising strong seedling, setting up populations with a high photosynthetic efficiency for increasing the supply of source, and improving the field management in filling stages to the duration of supply of photosynthate to grains.

A genetic linkage map of the apple, composed of 175 SSR and 105 SRAP markers, has been constructed using 110 F1 individuals obtained from a cross between the ‘Red Fuji’ Malus domestica and ‘Hongrou’ Malus sieversii cultivars, which have relatively high levels of DNA marker polymorphism and differ remarkably in fruit-related traits. The linkage map comprised 17 linkage groups, covering 1299.67 cM with an average marker distance of 4.6 cM between adjacent markers, or approximately 91% of Malus genome. Linkage groups were well populated and, although marker density ranged from 2.1 to 9.5 cM, just 10 gaps of more than 15 cM were observed. Moreover, just 12.5% of markers displayed segregation distortion. The present genetic linkage map was used to identify quantitative trait loci (QTLs) affecting fruit-related traits. 23 QTLs for ten fruit traits were detected by multiple interval mapping: 3 QTLs for Vc content, One QTL for single fruit weight, 2 QTLs for peel-phenols content, 2 QTLs for flesh-hardness, 2 QTLs for diameter, 6 QTLs for acid content, 1 QTL for sugar content, 2 QTLs for soluble solids content, 2 QTLs for flesh-phenols and 2 QTLs for brittleness. These QTLs were located on linkage groups C1, C2, C3, C5, C6, C7, C9, C10, C14 and C17, respectively. The phenotypic variations exhibited by each QTL ranged from 2% to 72%, and their LOD values varied from 2.03 to 8.93, of which five QTLs were major effect genes (R2 ≥ 10%). The tight linkage markers (*me2em7-460f, *MS01a03-180m, *me1em6-307m, *CH05c06-102f, *me1em8-423f) would be helpful to elucidate the molecular mechanisms of apple domestication and breeding in the future.

The present study focuses on the effects and suggests possible mechanisms of the traditional Chinese medicine Dilong as compared to dexamethasone on lower respiratory tract remodeling in rats with asthma. The number of leukocytes and eosinophils in blood from the inferior vena cava and bronchoalveolar lavage fluid (BALF) were counted. Lung tissues underwent hematoxylin and eosin staining. The thickness of the basement membrane and smooth muscle or the airways, and the ratio of inner to outer diameter of the airway wall were measured. Levels of transforming growth factor β1 (TGF-β1), matrix metallopeptidase 9/tissue inhibitor of metalloproteinase 1 (MMP-9/TIMP-1), urokinase plasminogen activator (uPA), plasminogen activator inhibitor 1 (PAI-1), and c-Myc(mRNA) were evaluated. Results indicate that treatment with Dilong decreased the number of eosinophils in blood and BALF, decreased levels of TGF-β1, MMP-9/TIMP-1, uPA, PAI-1 and c-Myc, and ameliorated the thickening of airway walls, airway basement membrane and airway smooth muscle. Co-treatment with dexamethasone was found to intensity these effects. The cellularity of eosinophils and thickness of the airway basement membrane and smooth muscle were positively correlated with levels of TGF- 1, uPA, and c-Myc. Treatment with Dilong, either alone or in combination with dexamethasone, could inhibit and partly reverse airway remodeling in rats with asthma at an early stage.

This paper investigated the effect of the application of sewage sludge on the growth rates and absorption rates of Pb and As in potted water spinach. Our results indicated that application of sewage sludge promoted vegetable growth, and the dry weight of water spinach reached a maximal value (4.38 ± 0.82 g) upon 8% sludge application. We also found that the dry weights of water spinach after treatment were all greater than those of the control systems (CK). Treatment with sludge promoted the absorption of Pb and As in water spinach, with a significant (p < 0.05) increase of absorbed Pb following treatment concentrations above 10%, and a peak absorption of As at 8%. Finally, we found that concentrations of Pb and As were higher in rhizosphere-attached soil than in free pot.

In this study, we investigated the therapeutic effects of Human Mesenchymal Stem Cells (hMSCs) cultured by hanging drop and conventional culturing methods on cerebellar repair in hypoxic-ischemic (HI) brain injured mice. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to analyze the expression levels of three stemness genes, Oct4, Sox2 and Nanog, and the migration related gene CXCR4. MSC prepared by hanging drop or conventional techniques were administered intranasally to nine day old mice, and analyzed by MRI at day 28. Results indicate that the MSCs, especially the hanging drop cultured MSCs, significantly improved the mice’s cerebellar damage repair. MSCs derived from the hanging drop culture were smaller than those from the conventional culture. The gene expression levels were significantly increased for the MSCs derived from the hanging drop culture. The mechanism might relate to the fact that the hanging drop cultured MSCs can be kept in an undifferentiated state, resulting in its higher expression level of migration receptor of CXCR4.