Analysis of genetic diversity in Arrhenatherum elatius Germplasm using inter-simple sequence repeat (ISSR) markers

The genetic diversity of 19 Arrhenatherum elatius accessions was analyzed using 100 inter-simple sequence repeat (ISSR) primers, out of which 11 generated distinct amplification products. Out of the 152 total bands detected, 107 were polymorphic. The percentage of polymorphic bands (PPB) was 68.9% with an average of 9.73 polymorphic bands per primer. The ISSR-based genetic similarity (GS) coefficients among the 19 accessions ranged from 0.4821 to 0.7411, revealing high genetic diversity. Based on the UPGMA cluster analysis and the principal components analysis (PCA), the 19 A. elatius accessions were divided into three groups with similar situations. We found that the genetic distance was related to the geographical distance among the 19 A. elatius accessions studied. These results confirm the potential value of genetic diversity preservation for future breeding programs.Key words: Arrhenatherum elatius, genetic diversity, inter-simple sequence repeat (ISSR) markers

Growth and physiological responses to water and nutrient stress in oil palm

The research was conducted to detect changes in growth, physiology and nutrient concentration in response to two watering regimes (well-watered and water-stress conditions) and to two nutrient regimes (with or without fertilization) of oil palm. Under stress conditions, changes in plant growth, dry matter allocation, relative water content, leaf relative conductivity, leaf N, P and K concentration are usually observed. These characteristics and related parameters were determined and the experiment results are listed as follows: (1) fertilization promoted the growth of oil palm under well-watered conditions, while under water stress conditions its effects on growth was negative. The ratio of root/shoot was increased under water stress condition; (2) relative water content and chlorophyll a/b content were gradually decreased while leaf relative conductivity was increased quickly under water and nutrient stress conditions during the experiment. It is obvious that water stress had a greater influence than nutrient stress on these parameters; (3) water and nutrient stress decreased leaf nitrogen and phosphorus concentration but increased potassium concentration; the combination of water and nutrient stress made significant effects on nitrogen and phosphorus concentration, but no significant effects on potassium concentration. Moreover, deficiency of both water and nutrients in combination had the greatest impact on changes in these traits of oil palm.Key words: Plant growth, physiology response, nutrient concentration, water stress, nutrient stress

Development of a Broad-Spectrum Antiviral Agent with Activity Against Herpesvirus Replication and Gene Expression

Purpose: To evaluate the broad-spectrum antiviral activity of peptide H9 (H9) in vitro in order to gain insight into its underlying molecular mechanisms.Method: Antiviral activity against Herpes simplex virus type 1 (HSV-1) was determined using thiazolyl blue (MTT) assay. Polymerase Chain Reaction (PCR) was employed to assay H9 antiviral activity against human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The inhibitory effect of H9 on the replication of these viral genes including early genes was assayed by real time-Ppolymerase chain reaction (RT-PCR) and Western blot.Results: H9 possessed significant inhibitory effect on the four different herpesviruses with 50 % inhibitory concentration (IC50) of 1.21 ng/mL (HSV-1). AD169 infection was strongly inhibited with an EC50 value of 0.46 ng/ml. The anti-herpesviral activity of H9 was dose-dependent. The peptide acted primarily during the early stage of infection by detection of the early genes.Conclusion: The results demonstrate that H9 can inhibit the infection of HSV-1, EBV and HCMV. Furthermore, H9 has a broad-spectrum anti-herpesviral effect in vitro based on targeted killing of infected cells expressing genes.Keywords: Antagonist, Trapping receptor/ligand, Broad-spectrum, Anti-herpesvirus, H9 peptide, Gene expressio

Efficient inverted polymer solar cells with thermal-evaporated and solution-processed small molecular electron extraction layer

Efficient inverted polymer solar cell is reported upon by integrating with a small molecular 1,3,5-tri(phenyl-2-benzimi-dazolyl)-benzene (TPBi) electron extraction layer (EEL) at low processing temperature with thermal-evaporation and solution-process, resulting in the power conversion efficiencies of 3.70 and 3.47, respectively. The potential of TPBi as an efficient EEL is associated with its suitable electronic energy level for electron extraction and hole blocking from the active layer to the indium tin oxide cathode. © 2013 American Institute of Physics

Physical mapping of a powdery mildew resistance related gene Hv-S/TPK by FISH with a TAC clone in wheat

Dissertação de mestrado integrado em Medicina (Hematologia), apresentado á Faculdade de Medicina da Universidade de Coimbra.A Policitemia Vera (PV) é uma doença clonal de etiologia desconhecida, na maior parte dos casos, que envolve a célula estaminal progenitora hematopoiética multipotencial. É uma neoplasia mieloproliferativa crónica (NMP) que se caracteriza pela expansão das três linhas celulares hematopoiéticas: eritróide, granulocítica e megacariocítica, com predomínio da primeira, de modo independente dos mecanismos normais de regulação da eritropoiese. Além disso, as células têm aspecto morfológico normal, a fibrose medular é pouco significativa e os níveis de eritropoietina (Epo) são habitualmente normais a baixos. Além da hipercelularidade medular com sobreprodução de uma ou de todas as linhas celulares, a doença cursa com hematopoiese extramedular, hiperviscosidade, propensão para complicações como trombose ou hemorragia e risco de desenvolvimento de mielofibrose ou transformação em leucemia aguda. A descrição relativamente recente da associação de uma mutação no gene JAK2, localizado no cromosoma 9p24, com as doenças mieloproliferativas clássicas negativas para BCR-ABL, como a PV, veio permitir avanços significativos na compreensão da patofisiologia deste grupo de doenças hematológicas. A mutação provoca uma alteração do aminoácido V (valina) para F (fenilalanina) na posição 617 (JAK2V617F). De acordo com os dados publicados, a frequência da detecção da mutação JAK2V617F em doentes com PV é de cerca de 95%. A proteína JAK2 é uma tirosina cinase citoplasmática, que se encontra associada ao domínio intracelular dos receptores de citocinas (como a Epo e trombopoietina - Tpo), e de factores de crescimento, essenciais para a função destes receptores. A mutação da JAK2 conduz à activação constitutiva dos receptores, independente da ligação à respectiva citocina e/ou hipersensibilidade a factores de crescimento, com consequente activação de múltiplas vias de sinalização intracelulares como a JAK/STAT (Janus Kinase/Signal Transductor and activator of transcription), a PI3K (fosfatidilinositol 3 cinase) e a MAPK (proteína cinase activadora de mitose), envolvidas na transformação e proliferação dos progenitores hematopoiéticos. Por outro lado, as células evidenciam alteração na diferenciação terminal e resistência à apoptose in vitro que poderá estar relacionada com o aumento da expressão da proteína anti-apoptótica Bcl-XL. Além dos avanços no diagnóstico, a detecção da mutação JAK2V617F tem contribuido para melhorar a classificação e a terapêutica dos doentes com PV. Deste modo, o conhecimento dos mecanismos moleculares envolvidos na PV tem levado os investigadores à descoberta de novos fármacos dirigidos ao defeito molecular, permitindo novas abordagem terapêuticas mais eficazes e provavelmente de menor toxicidade. Este trabalho procura fazer uma revisão sobre o actual conhecimento da caracterização molecular e clínica da PV e quais as suas implicações no diagnóstico e abordagem terapêutica desta NMP.Polycythemia Vera (PV) is a clonal disease of unknown etiology, which often involves the pluripotential hematopoietic stem cell. This disease integrates the family of chronic myeloproliferative neoplasm (MPN) and is characterized by the growth of the three hematopoietic celular lineages: granulocytic, megakaryocytic and erythroid, with predominance of the last one and regardless the normal mechanisms of erythropoiesis regulation. Moreover, cells have normal morphological aspect, bone marrow shows slight fibrosis and the levels of erythropoietin (Epo) usually vary from normal to low. Besides marrow hypercellularity with overproduction of one or all the celular lineages, the disease courses with extramedullary hematopoiesis, hyperviscosity, leading to complications such as thrombosis or bleeding and risk of transformation to myelofibrosis or acute leukemia. Recently it has been described the association between the mutation in the JAK2 gene, located on chromosome 9p24, with the classic myeloproliferative disorders BCR-ABL negative, such as PV, which has brought significant advances in the understanding of the pathophysiology of this group of hematologic malignancies. The mutation causes a change of amino acid V (valine) to F (phenylalanine) at position 617 (JAK2V617F). According to published data, the frequency of JAK2V617F mutation detected in patients with PV is about 95%. JAK2 protein is a cytoplasmic tyrosine kinase, which is associated to the intracelular domain of cytokine receptors, such as Epo and thrombopoietin (Tpo), and growth factors which are essential to the function of these receptors. JAK2 mutation leads to the constitutive receptors activation, independent of connection to their cytokine and / or hypersensitivity to growth factors, with consequent activation of multiple intracellular signaling pathways such as JAK / STAT (Janus Kinase / Signal transducer and transcription activator), the PI3K (phosphatidylinositol 3 kinase) and MAPK (Mitogen-activated protein), involved in the transformation and proliferation of hematopoietic progenitors. Moreover, the cells show changes in terminal differentiation and resistance to in vitro apoptosis which is possibly related to the increasing expression of anti-apoptotic protein Bcl-XL. In addition to the advances in diagnosis, detection of JAK2V617F mutation has contributed to the improvement of classification and treatment in patients with PV. Thus, knowledge of the molecular mechanisms involved in PV has led investigators to the discovery of new drugs targeting molecular defects, allowing new therapeutic approach more efficient and probably less toxic. The aim of this article is to review the current knowledge of clinical and molecular characterization of PV, and its implications on the diagnosis and therapeutic approach of this myeloproliferative disorder

Anti-Tumor Effect of Cactus Polysaccharides on Lung Squamous Carcinoma Cells (SK-MES-1)

Background: Cactus polysaccharides are the active components of Opuntia dillenii which have been used extensively in folk medicine. In thisstudy, we investigate the anti-tumor effect of cactus polysaccharides on lung squamous carcinoma cells SK-MES-1.Materials and Methods: The inhibitory effect of Cactus polysaccharides on lung squamous carcinoma cells were detected by MTT assay. Cellcycle was determined by flow cytometry and cell apoptosis was determined by AnnexinV assay. Western-blotting was applied to detect P53 andPTEN protein expression in the cells treated with cactus polysaccharides.Results: Results showed that different concentrations of wild cactus polysaccharides prevent SK-MES-1 cells growth and induces S phase arrest.The data also revealed that cactus polysaccharides cause apoptosis in SK-MES-1 cells determined by Annexin-V assay. Furthermore, cactuspolysaccharides induced growth arrest and apoptosis may be due to the increase of P53 and phosphatase and tension homolog deleted onchromosome ten (PTEN) protein.Conclusion: Cactus polysaccharides have anti-tumor activity on lung squamous carcinoma cells.Key words: Cactus polysaccharides, Lung squamous carcinoma, Anti-tumor effect, P53, PTEN Abbreviations: PTEN :phosphatase and tension homolog deleted on chromosome ten; NSCLC: Non-small-cell lung cancer; FBS :Phosphate buffered saline; MTT:3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; PBS: Phosphate buffered saline; DMSO:Dimethyl sulfoxide; PI: Propidium iodide

Kinase inhibit region of SOCS3 attenuates IL6-induced proliferation and astrocytic differentiation of neural stem cells via cross talk between signaling pathways

Aims: Efficiency of neural stem cells (NSCs) therapy for brain injury is restricted by astrogliosis around the damaged region, in which JAK2/STAT3 signaling plays a key role. The SOCS3 that can directly inhibit JAK/STAT3 pathway. Here, we investigated the effects of a fusion peptide that combined kinase inhibitory region (KIR) of SOCS3 and virus trans-activator of transcription (TAT) on biological behavior of cultured NSCs under inflammatory conditions. Methods: NSCs were isolated from embryonic brain of SD rats, TAT-KIR was synthesized, and penetration rate was evaluated by flow cytometry (FACS). CCK8, immunostaining, and FACS were used to detected of TAT-KIR on the proliferation of NSCs. The expressions of GFAP and β tubulin III positive cells induced by IL6 with/without TAT-KIR were examined by immunostaining and Western blotting to observe the NSCs differentiation, and the effect of TAT-KIR on signaling cross talk was observed by Western blotting. Results: Penetration rate of TAT-KIR into primary cultured NSCs was up to 94%. TAT-KIR did not affect the growth and viability of NSCs. It significantly reduced the NSCs proliferation that enhanced by IL-6 stimulation via blocking the cell cycle progression from the G0/G1 to S phase. In addition, TAT-KIR attenuated astrocytic differentiation and kept high level of neuronal differentiation derived from IL-6-induced NSCs. The fate of NSCs differentiation under inflammatory conditions was affected by TAT-KIR, which was associated with synchronous inhibition of STAT3 and AKT, while promoting JNK expression. Conclusion: TAT-KIR mimetic of SOCS3 could be a promising approach for brain repair via regulating the biological behaviors of exogenous NSCs

A genetic contribution from the Far East into Ashkenazi Jews via the ancient Silk Road

Contemporary Jews retain a genetic imprint from their Near Eastern ancestry, but obtained substantial genetic components from their neighboring populations during their history. Whether they received any genetic contribution from the Far East remains unknown, but frequent communication with the Chinese has been observed since the Silk Road period. To address this issue, mitochondrial DNA (mtDNA) variation from 55,595 Eurasians are analyzed. The existence of some eastern Eurasian haplotypes in eastern Ashkenazi Jews supports an East Asian genetic contribution, likely from Chinese. Further evidence indicates that this connection can be attributed to a gene flow event that occurred less than 1.4 kilo-years ago (kya), which falls within the time frame of the Silk Road scenario and fits well with historical records and archaeological discoveries. This observed genetic contribution from Chinese to Ashkenazi Jews demonstrates that the historical exchange between Ashkenazim and the Far East was not confined to the cultural sphere but also extended to an exchange of genes