Purification and characterization of an intracellular chymotrypsin-like serine protease from Thermoplasma volcanium

An intracellular serine protease produced by Thermoplasma (Tp.) voleanium was purified using a combination of ammonium sulfate fractionation, ion exchange, and et-casein agarose affinity chromatography. This enzyme exhibited the highest activity and stability at pH 7.0, and at 50 degrees C. The purifed enzyme hydrolyzed synthetic peptides preferentially at the carboxy terminus of phenylalanine or leucine and was almost completely inhibited by PMSF, TPCK, and chymostatin, similarly to a chymotrypsin-like serine protease. Kinetic analysis of the Tp. volcanium protease reaction performed using N-succinyl-L-phenylalanine-p-nitroanilide as substrate revealed a K. value of 2.2 mm and a V-max value of 0.045 mu mol(-1) m1(-1) min(-1). Peptide hydrolyzing activity was enhanced by > 2-fold in the presence of Ca2+ and Mg2+ at 2-12 mm concentration. The serine protease is a monomer with a molecular weight of 42 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and zymogram activity staining

The left sternalis muscle variation detected during mastectomy

The sternalis muscle variation is a well-known anatomical situation. It is present in 8.7% of women and 6.4% of men, although the incidence varies according to sex, race and ethnicity. During a left modified radical mastectomy operation on a 46-year-old female patient a sternalis muscle was detected on the pectoralis major muscle in the superficial fascia. It was in craniocaudal position and was parallel to the body of the sternum. The cylindrical muscle was approximately 8 cm in length and 2 cm in diameter. Such variations are considered to have their origin in embryological development. Awareness of muscular variations and their identification is important both for procedure through the proper dissection planes during breast surgery and in radiological examination and follow-up

Extracción en fase sólida de β-sitosterol y α-tocoferol de destilados de aceite de girasol desodorizado utilizando zeolita desilicada

In this study, the efficiency of using zeolite-based adsorbents in a solid phase extraction (SPE) procedure of α-tocopherol and β-sitosterol isolation from Sunflower Oil Deodorizer Distillate (SuDOD) with­out pre-treatment was investigated. The results showed that 99.2% α-tocopherol and 97.3% β-sitosterol were suc­cessfully isolated as pure fractions from SuDOD, when desilicated ZSM-5-type zeolite (DSiZSM-5) was used as adsorbent on a SPE. A simple and rapid HPLC method for simultaneous α-tocopherol and β-sitosterol analysis was developed and validated according to AOAC guidelines. It was found that the inclusion of a DSiZSM-5 SPE step increased the precision of the α-tocopherol and β-sitosterol analysis. In conclusion, DSiZSM-5 zeo­lite was proven to be an efficient adsorbent which can be used not only for the recovery of α-tocopherol and β-sitosterol from SuDOD in industrial scale, but also in a laboratory scale clean-up method prior to the analysis of α-tocopherol and β-sitosterol.En este estudio, se investigó la eficacia del uso de adsorbentes a base de zeolita en el procedimiento de extracción en fase sólida (EFS) para el aislamiento de α-tocoferol y β-sitosterol a partir de destilados de aceites de girasol desodorizados (SuDOD) sin ningún tratamiento previo. Los resultados mostraron que el 99,2% de α-tocoferol y el 97,3% de β-sitosterol se aislaron con éxito como fracciones puras de SuDOD, cuando se usó zeolita de tipo ZSM-5 desilicado (DSiZSM-5) como adsorbente en una EFS. Se desarrolló y validó un método HPLC simple y rápido para el análisis simultáneo de α-tocoferol y β-sitosterol de acuerdo con las directrices de la AOAC. Se encontró que la inclusión del paso DSiZSM-5 EFS aumentó la precisión del análisis de α-tocoferol y β-sitosterol. En conclusión, se demostró que la zeolita DSiZSM-5 es un adsorbente eficiente que puede usarse, no solo para la recuperación de α-tocoferol y β-sitosterol de SuDOD a escala industrial, sino también en un método de limpieza a escala de laboratorio antes del análisis de α-tocoferol y β-sitosterol

High resolution mapping of Twist to DNA in Drosophila embryos: Efficient functional analysis and evolutionary conservation

Cis-regulatory modules (CRMs) function by binding sequence specific transcription factors, but the relationship between in vivo physical binding and the regulatory capacity of factor-bound DNA elements remains uncertain. We investigate this relationship for the well-studied Twist factor in Drosophila melanogaster embryos by analyzing genome-wide factor occupancy and testing the functional significance of Twist occupied regions and motifs within regions. Twist ChIP-seq data efficiently identified previously studied Twist-dependent CRMs and robustly predicted new CRM activity in transgenesis, with newly identified Twist-occupied regions supporting diverse spatiotemporal patterns (>74% positive, n = 31). Some, but not all, candidate CRMs require Twist for proper expression in the embryo. The Twist motifs most favored in genome ChIP data (in vivo) differed from those most favored by Systematic Evolution of Ligands by EXponential enrichment (SELEX) (in vitro). Furthermore, the majority of ChIP-seq signals could be parsimoniously explained by a CABVTG motif located within 50 bp of the ChIP summit and, of these, CACATG was most prevalent. Mutagenesis experiments demonstrated that different Twist E-box motif types are not fully interchangeable, suggesting that the ChIP-derived consensus (CABVTG) includes sites having distinct regulatory outputs. Further analysis of position, frequency of occurrence, and sequence conservation revealed significant enrichment and conservation of CABVTG E-box motifs near Twist ChIP-seq signal summits, preferential conservation of ±150 bp surrounding Twist occupied summits, and enrichment of GA- and CA-repeat sequences near Twist occupied summits. Our results show that high resolution in vivo occupancy data can be used to drive efficient discovery and dissection of global and local cis-regulatory logic

Structural, thermal and magnetic properties of barium-ferrite powders substituted with Mn, Cu or Co and X (X = Sr and Ni) prepared by the sol-gel method

In this study, Ferrite A (undoped barium hexaferrite), Ferrite B (MnCuNi-doped barium hexaferrite), Ferrite C (MnCuSr-doped barium hexaferrite), Ferrite D (MnCoNi-doped barium hexaferrite) and Ferrite E (MnCoSr-doped barium hexaferrite) powders were prepared by sol-gel processing. The produced powders were calcined at 550 °C for 6 h and sintered at 1000 °C for 5 h to obtain the required phases. The powders were characterized by differential thermal analysis/thermogravimetric analysis (DTA/TG), X-ray diffractometry (XRD) and scanning electron microscopy (SEM), and vibrating-sample magnetometry (VSM). The XRD patterns indicated that the pure barium ferrite phase was not obtained. The presence of M-type BaFe 11.6Mn 0.4O 19 was confirmed in the Ferrite B and Ferrite D patterns. In the Ferrite C pattern, there were the phases of BaFe 12O 19, Ba 2Cu 2Fe 12O 22 (X or Z-type) and Sr 3Fe 2O 6.16. The Ba 0.5Sr 0.5Fe 12 phase was easily observed in the Ferrite E pattern. The results showed that the dopant materials significantly change the particle shape of Ferrite A powders, but also lower the value of the coercivity. A higher saturation magnetization was observed for the Ferrite D powder

Chromogenic and Fluorogenic Sensing of Biological Thiols in Auqeous Solutions Using BODIPY-Based Reagents

Cataloged from PDF version of article.udicious design of BODIPY dyes carrying nitroethenyl substituents in conjugation with the BODIPY core yields dyes that respond to biological thiols by both absorbance and emission changes. Incorporation of solubilizing ethyleneglycol units ensures water solubility. The result is bright signaling of biologically relevant thiols in the longer wavelength region of the visible spectrum and in aqueous solutions

A practical probabilistic earthquake hazard analysis tool: case study Marmara region

Earthquakes have a damaging impact on the economic welfare and resilience of communities, particularly in developing countries. Seismic hazard assessment is the first step towards performing prevention, preparedness, and response or recovery actions to reduce seismic risk. This paper presents a computation tool for predicting the seismic hazard at the macro level as a part of a comprehensive multi-hazard framework on earthquake risk assessment. The probabilistic seismic hazard analysis (PSHA) procedure is based on the Monte-Carlo approach, and particular attention is paid to the definition of source zones assigned in the study area. Both Poisson and time dependent (renewal) models are adopted to quantify the effect of temporal dependencies between seismic events, while near-field rupture directivity effects are also taken into account. Marmara region in Turkey is selected as a case study area to perform a new seismic hazard analysis and verify the accuracy of the proposed tool. The results show good agreement with results from the recent SHARE project and the latest Turkish Earthquake Design code hazard maps. This confirms that the proposed PSHA method can be an attractive alternative to the direct integration based methods due to its practicality and powerful handling of uncertainties