Identification of Sare0718 As an Alanine-Activating Adenylation Domain in Marine Actinomycete Salinispora arenicola CNS-205

BACKGROUND: Amino acid adenylation domains (A domains) are critical enzymes that dictate the identity of the amino acid building blocks to be incorporated during nonribosomal peptide (NRP) biosynthesis. NRPs represent a large group of valuable natural products that are widely applied in medicine, agriculture, and biochemical research. Salinispora arenicola CNS-205 is a representative strain of the first discovered obligate marine actinomycete genus, whose genome harbors a large number of cryptic secondary metabolite gene clusters. METHODOLOGY/PRINCIPAL FINDINGS: In order to investigate cryptic NRP-related metabolites in S. arenicola CNS-205, we cloned and identified the putative gene sare0718 annotated "amino acid adenylation domain". Firstly, the general features and possible functions of sare0718 were predicted by bioinformatics analysis, which suggested that Sare0718 is a soluble protein with an AMP-binding domain contained in the sequence and its cognate substrate is L-Val. Then, a GST-tagged fusion protein was expressed and purified to further explore the exact adenylation activity of Sare0718 in vitro. By a newly mentioned nonradioactive malachite green colorimetric assay, we found that L-Ala but not L-Val is the actual activated amino acid substrate and the basic kinetic parameters of Sare0718 for it are K(m) = 0.1164±0.0159 (mM), V(max) = 3.1484±0.1278 (µM/min), k(cat) = 12.5936±0.5112 (min(-1)). CONCLUSIONS/SIGNIFICANCE: By revealing the biochemical role of sare0718 gene, we identified an alanine-activating adenylation domain in marine actinomycete Salinispora arenicola CNS-205, which would provide useful information for next isolation and function elucidation of the whole cryptic nonribosomal peptide synthetase (NRPS)-related gene cluster covering Sare0718. And meanwhile, this work also enriched the biochemical data of A domain substrate specificity in newly discovered marine actinomycete NRPS system, which bioinformatics prediction will largely depend on

Evaluaci?n de la disponibilidad de biomasa y capacidad de rebrote de le?osas forrajeras en potreros del tr?pico seco de Nicaragua

Tesis (M. Sc) -- CATIE, Turrialba (Costa Rica), 2012El prop?sito del presente estudio es conocer la capacidad de rebrote y los rasgos funcionales que explican la productividad de biomasa en seis especies forrajeras arbustivas en potrero del tr?pico seco de Nicaragua. Finalmente las especies con mayor capacidad de rebrote y Producci?n de biomasa se asocian con una menor AF, mientras que el AFE no fue un rasgo explicativo. Es importante mencionar que la selecci?n de especies forrajeras arbustivas requieren de criterios m?s amplios donde se consideren rasgos funcionales de las plantas que mejoren la plasticidad fenot?pica de las especies, por ejemplo el tama?o de la hoja, la dureza de la hoja, la relaci?n hoja - tallo dentro de las fracciones comestibles, rasgos nutricionales de las especies como prote?na cruda (PC), fibra neutro detergente (FNA), fibra acido detergente (FAD), lignina y taninos), reservas de carbohidratos en tallos, ra?ces finas y estructurales y finalmente los efectos de la poda en los diferentes niveles de manejo y estaci?n clim?tica. To contribute to the search for these resources we attempt to resprouting ability of six forage trees and shrubs and measure their functional traits associated with biomass productivity. These species were selected taking into account knowledge regarding species with the greatest potential, preference tests performed with livestock and abundance in pastures. Finally those with high coping mechanisMON to coppicing and biomass production are associated with lower LA, while the SLA was not an explanatory feature. It is worth noting that the selection of forage shrubs needs broader criteria which consider plant functional traits that enhance the phenotypic plasticity of the species, such as leaf size, leaf toughness, leaf - stem fractions in edible nutritional traits of species such as crude protein (CP), neutral detergent fiber (FNA), acid detergent fiber (ADF), lignins and tannins), carbohydrate reserves in steMON and, roots, and the structural the effects of pruning on the different levels of management and seasonal patterns

De figurantes a atores: o coletivo na luta das famílias dos autistas

This paper deals with the formation of collective identities focusing on the families of people with autism. The arrival of autism is determinative of a radical change in everyday life, social relations and expectations of the families, yet it becomes a factor that drives them to the organization and struggle for equal living conditions and fulfillment of their rights. Stigma, shared memories and struggles are presented as the foundation of their social identity. Based on the theoretical tenors of Tajfel, Goffman and Martin-Baró the article seeks to highlight that the families emerge as social actors when mobilizing and addressing their claims to the public.Cet article traite de la formation des identités collectives des familles de personnes atteintes d'autisme. L'arrivée de l'autisme détermine le changement radical de la vie quotidienne, des relations sociales et les attentes des familles, cependant, il devient un facteur qui pousse l'organisation et la lutte pour l'égalité des conditions de vie et pour la réalisation de leurs droits. La stigmatisation, la mémoire partagée et les luttes sont les éléments soulignés comme la base de l'identité sociale. En considérant les écrits de Tajfel, Goffman et Martín- Baró, cet article démontre que la mobilisation et l'exposition de leurs demandes font émerger des familles comme acteurs sociaux sur la scène publique.Este artigo trata da formação de identidades coletivas centrando-se nas famílias de pessoas com autismo. A chegada do autismo determina a mudança radical da vida cotidiana, das relações sociais e das expectativas dessas famílias, contudo, torna-se um fator que as impulsiona à organização e à luta pela igualdade de condições de vida e concretização de seus direitos. O estigma, a memória compartilhada e as lutas se apresentam como a base de sua identidade social. Valendo-se dos conteúdos teóricos de Tajfel, Goffman e Martín-Baró, o artigo procura evidenciar que a mobilização e a exposição de suas reivindicações fazem as famílias emergirem na cena pública como atores sociais.Este artículo se refiere a la formación de identidades colectivas, centrándose en el grupo de familias de personas con autismo. La llegada del autismo determina el cambio radical de la vida cotidiana, las relaciones sociales y las expectativas de las familias, sin embargo, se convierte en un factor que las impulsa hacia la organización, la lucha por la igualdad de condiciones de vida y la realización de sus derechos. El estigma, la memoria compartida y las luchas se destacan como fundamentos de la identidad social. Utilizando los contenidos teóricos de Tajfel, Goffman y Martín-Baró, el artículo intenta demostrar que al movilizarse y hacer públicas sus reclamaciones, las familias emergen como actores sociales

Sistem Informasi Akuntansi Potongan Pajak Pph 21 Menggunakan Visual Studio Pada PT. Yushindo Yasa Perkasa Ternate

Dalam hal ini peniliti melakukakan penelitian terhadap kegiatan administrasi Perusahaan, yang didalamnya terdapat kegiatan yang terkomputerisasi yang akan diusulkan menjadi sebuah aplikasi. Objek penelitian merupakan Perusahaan jasa konstruksi bangunan, jalan dan jembatan yang dalam pengelolaan data gaji dan pajak PPh pasal 21 belum terolah secara maksimal, walaupun telah terolah secara komputerisasi, sehingga perlu adanya suatu aplikasi sistem informasi yang dapat membantu Perusahaan mengolah data gaji dan perhitugan pajak penghasilan secara terintegrasi. Salah satu alat yang dapat mendukung hal tersebut adalah dengan pemakaian komputer dan sumber daya manusia yang memadai, sehingga dapat terbentuk sistem informasi yang baik. Tahap kedua yaitu analisis untuk mengetahui kelemahan- kelemahan dari sistem tersebut serta mengevaluasi hasilnya sehingga dapat melakukan perancangan sistem. Aplikasi bantu yang di gunakan adalah visual studio

Antiproliferative and palliative activity of flavonoids in colorectal cancer

Flavonoids are plant bioactive compounds of great interest in nutrition and pharmacology, due to their remarkable properties as antioxidant, anti-inflammatory, antibacterial, antifungal and antitumor drugs. More than 5000 different flavonoids exist in nature, with a huge structural diversity and a plethora of interesting pharmacological properties. In this work, five flavonoids were tested for their potential use as antitumor drugs against three CRC cell lines (HCT116, HT-29 and T84). These cell lines represent three different stages of this tumor, one of which is metastatic. Xanthohumol showed the best antitumor activity on the three cancer cell lines, even better than that of the clinical drug 5-fluorouracil (5-FU), although no synergistic effect was observed in the combination therapy with this drug. On the other hand, apigenin and luteolin displayed slightly lower antitumor activities on these cancer cell lines but showed a synergistic effect in combination with 5-FU in the case of HTC116, which is of potential clinical interest. Furthermore, a literature review highlighted that these flavonoids show very interesting palliative effects on clinical symptoms such as diarrhea, mucositis, neuropathic pain and others often associated with the chemotherapy treatment of CRC. Flavonoids could provide a double effect for the combination treatment, potentiating the antitumor effect of 5-FU, and simultaneously, preventing important side effects of 5-FU chemotherapy

Improved Production of Antifungal Angucycline Sch47554 by Manipulating Three Regulatory Genes in Streptomyces sp. SCC-2136

Sch47554 and Sch47555 are two angucyclines with antifungal activities against various yeasts and dermatophytes from Streptomyces sp. SCC‐2136. The schgene cluster contains several putative regulatory genes. Both schA4 and schA21were predicted as the TetR family transcriptional regulators, whereas schA16shared significant similarity to the AraC family transcriptional regulators. Although Sch47554 is the major product of Streptomyces sp. SCC‐2136, its titer is only 6.72 mg/L. This work aimed to increase the production of this promising antifungal compound by investigating and manipulating the regulatory genes in the Sch47554 biosynthetic pathway. Disruption of schA4and schA16 led to a significant increase in the production of Sch47554, whereas the titer was dramatically decreased when schA21 was disrupted. Overexpression of these genes gave opposite results. The highest titer of Sch47554 was achieved in Streptomyces sp. SCC‐2136/ΔschA4 (27.94 mg/L), which is significantly higher than the wild type. Our results indicate that SchA4 and SchA16 are repressors, whereas SchA21 acts as an activator. This work thus provides an initial understanding of functional roles of regulatory elements in the biosynthesis of Sch47554. Several efficient producing strains of Sch47554 were constructed by disrupting or over-expressing particular regulatory genes, which can be further engineered for industrial production of this medicinally important molecule