Laccase gene silencing negatively effects growth and development in Pleurotus ostreatus

42-51In this study we describe the effects of Pleurotus ostreatus transformants that have been silenced for laccase genes. Using the RNAi strategy, transformants with different levels of phenotypic alteration were obtained with respect to their oxidation capacity of 2,6-dimethoxyphenol (DMP). The analysis of laccase activity on DMP allowed us to select transformants with severe, medium and light phenotypic alteration in comparison to the PoB strain. The measurements of the average growth rate of the transformants with severe phenotypic alteration suggested that laccase expression could play a role in the vegetative growth of P. ostreatus. It was observed that both in solid and liquid cultures, PoB and the transformants express mRNA for lacc10, although the transformants with medium and severe phenotypic alteration present a decrease in intensity, especially in solid culture. This suggests that the product of this gene is responsible for the development of the mycelium and probably participates in the production of biomass in solid culture and also could be related to the decrease in the intensity of the constant isoenzyme observed in both culture systems

Structural stabilization of botulinum neurotoxins by tyrosine phosphorylation

AbstractTyrosine phosphorylation of botulinum neurotoxins augments their proteolytic activity and thermal stability, suggesting a substantial modification of the global protein conformation. We used Fourier-transform infrared (FTIR) spectroscopy to study changes of secondary structure and thermostability of tyrosine phosphorylated botulinum neurotoxins A (BoNT A) and E (BoNT E). Changes in the conformationally-sensitive amide I band upon phosphorylation indicated an increase of the α-helical content with a concomitant decrease of less ordered structures such as turns and random coils, and without changes in β-sheet content. These changes in secondary structure were accompanied by an increase in the residual amide II absorbance band remaining upon H-D exchange, consistent with a tighter packing of the phosphorylated proteins. FTIR and differential scanning calorimetry (DSC) analyses of the denaturation process show that phosphorylated neurotoxins denature at temperatures higher than those required by non-phosphorylated species. These findings indicate that tyrosine phosphorylation induced a transition to higher order and that the more compact structure presumably imparts to the phosphorylated neurotoxins the higher catalytic activity and thermostability

DD04107-Derived neuronal exocytosis inhibitor peptides: Evidences for synaptotagmin-1 as a putative target

The analgesic peptide DD04107 (Pal-EEMQRR-NH2) and its acetylated analogue inhibit a-calcitonin gene-related peptide (a-CGRP) exocytotic release from primary sensory neurons. Examining the crystal structure of the SNARE-Synaptotagmin-1(Syt1) complex, we hypothesized that these peptides could inhibit neuronal exocytosis by binding to Syt1, hampering at least partially its interaction with the SNARE complex. To address this hypothesis, we first interrogate the role of individual side-chains on the inhibition of a-CGRP release, finding that E1, M3, Q4 and R6 residues were crucial for activity. CD and NMR conformational analysis showed that linear peptides have tendency to adopt a-helical conformations, but the results with cyclic analogues indicated that this secondary structure is not needed for activity. Isothermal titration calorimetry (ITC) measurements demonstrate a direct interaction of some of these peptides with Syt1-C2B domain, but not with Syt7-C2B region, indicating selectivity. As expected for a compound able to inhibit a-CGRP release, cyclic peptide derivative Pal-E-cyclo[EMQK]R-NH2 showed potent in vivo analgesic activity, in a model of inflammatory pain. Molecular dynamics simulations provided a model consistent with KD values for the interaction of peptides with Syt1-C2B domain, and with their biological activity. Altogether, these results identify Syt1 as a potential new analgesic target. © 202

Estudio y caracterización de algunas 5-E-aldoximino-4-glucosilaminopirimidinas

La reacción de 1,6-dihidro-5-formil-4-β-D-(2 ', 3 ' 4 ', 6 '-tetra-O-acetil)-glucopiranosilamino-1-metil-2-metoxi-6-oxopirimidina con exceso de ClH.H2NOH permite aislar 5-E-aldoximino derivado como único producto. La estructura del mismo ha sido establecida en base a pruebas químicas y espectroscópicas.The reaction between 1,6-dihydro-S-formil-4- β -D-(tetra-O-acetyl}glucopyranosylamino-l-methyl-2-methoxy-6-oxo-pyrimidine and an excess of ClH.H2 NOH yields S-E-aldoximine derivative as the only producto On the basis of its chemical behaviour and espectroscopic data we propose its structure

Estudio y caracterización de algunas 5-E-aldoximino-4-glucosilaminopirimidinas

The reaction between 1,6-dihydro-5-formil-4-β-D-(tetra-O-acetyl)glucopyranosylamino-1-methyl-2-methoxy-6-oxo-pyrimidine and an excess of ClH.H2NOH yields 5-E-aldoximine derivative as the only product. On the basis of its chemical behaviour and espectroscopic data we propose its structure.La reacción de 1,6-dihidro-5-formil-4-β-D-(2 ', 3 ' 4 ', 6 '-tetra-O-acetil)-glucopiranosilamino-1-metil-2-metoxi-6-oxopirimidina con exceso de ClH.H2NOH permite aislar 5-E-aldoximino derivado como único producto. La estructura del mismo ha sido establecida en base a pruebas químicas y espectroscópicas

Neonate Human Remains: A Window of Opportunity to the Molecular Study of Ancient Syphilis

Ancient DNA (aDNA) analysis can be a useful tool in bacterial disease diagnosis in human remains. However, while the recovery of Mycobacterium spp. has been widely successful, several authors report unsuccessful results regarding ancient treponemal DNA, casting doubts on the usefulness of this technique for the diagnosis of ancient syphilis. Here, we present results from an analysis of four newborn specimens recovered from the crypt of “La Ermita de la Soledad” (XVI–XVII centuries), located in the province of Huelva in the southwest of Spain. We extracted and analyzed aDNA in three independent laboratories, following specific procedures generally practiced in the aDNA field, including cloning of the amplified DNA fragments and sequencing of several clones. This is the most ancient case, reported to date, from which detection of DNA from T. pallidum subspecies pallidum has been successful in more than one individual, and we put forward a hypothesis to explain this result, taking into account the course of the disease in neonate individuals

Spanish ATLAS tier-2: Facing up to LHC Run 2

The goal of this work is to describe the way of addressing the main challenges of Run 2 by the Spanish ATLAS Tier-2. The considerable increase of energy and luminosity for the upcoming Run 2 with respect to Run 1 has led to a revision of the ATLAS computing model as well as some of the main ATLAS computing tools. In this paper, the adaptation to these changes will be described. The Spanish ATLAS Tier-2 is a R&D project which consists of a distributed infrastructure composed of three sites and its members are involved in ATLAS computing progress, namely the work in different tasks and the development of new tools (e.g. Event Index)This work has been supported by MINECO, Spain (Proj. Ref. FPA2010-21919-C03-01,02,03 & FPA2013-47424-C3,01,02,03), which include FEDER funds from the European Unio

Functional upgrading in China’s export processing sector

Functional upgrading occurs when a firm acquires more sophisticated functions within an existing value chain. In this paper, we analyze if there is evidence of this type of upgrading in China’s export processing regime by investigating dynamics in the relative prevalence of Import & Assembly (IA) versus Pure Assembly (PA) processing trade over the period 2000-2013. Firms in both regimes provide similar manufacturing services to foreign companies, but IA firms also conduct the sophisticated tasks of quality control, searching, financing and storing imported materials. Consistent with a trend of functional upgrading, we show that the share of IA trade in total processing trade has increased rapidly during the period 2000-2006, both overall and within product categories. Furthermore, we find that this trend has gone hand in hand with improvements in a sector’s labor productivity and unit values. Against expectations, we find that this process has slowed down notably during the period 2006-2013.status: publishe

Primary retroperitoneal mucinous cystadenocarcinoma: report of two cases

BACKGROUND: Retroperitoneal cystadenocarcinomas are rare lesions, the majority of cases presented as one-patient reports. METHODS: We present two cases of retroperitoneal cystadenocarcinoma, both in women of reproductive age: one with aggressive behavior, and the remaining case, with a more indolent clinical evolution. RESULTS: One case presented as pelvic tumor, was treated with surgical resection of the disease, but manifested with recurrent disease a few months later despite use of chemotherapy. The second case involved a patient with diagnosis of abdominal tumor; during laparotomy, a retroperitoneal tumor was found and was totally removed. At follow-up, the patient is disease-free with no other treatment. CONCLUSION: The behavior and treatment of retroperitoneal cystadenocarcinoma are controversial. We suggest aggressive surgery including radical hysterectomy and bilateral salpingoopherectomy with adjuvant chemotherapy in these cases