Pyrolysis of peptides and proteins. Analytical study and environmental applications

Analytical pyrolysis was used to investigate the formation of diketopiperazines (DKPs) which are cyclic dipeptides formed from the thermal degradation of proteins. A quali/quantitative procedure was developed combining microscale flash pyrolysis at 500 °C with gas chromatography-mass spectrometry (GC-MS) of DKPs trapped onto an adsorbent phase. Polar DKPs were silylated prior to GC-MS. Particular attention was paid to the identification of proline (Pro) containing DKPs due to their greater facility of formation. The GC-MS characteristics of more than 80 original and silylated DKPs were collected from the pyrolysis of sixteen linear dipeptides and four model proteins (e.g. bovine serum albumin, BSA). The structure of a novel DKP, cyclo(pyroglutamic-Pro) was established by NMR and ESI-MS analysis, while the structures of other novel DKPs remained tentative. DKPs resulted rather specific markers of amino acid sequence in proteins, even though the thermal degradation of DKPs should be taken into account. Structural information of DKPs gathered from the pyrolysis of model compounds was employed to the identification of these compounds in the pyrolysate of proteinaceous samples, including intrinsecally unfolded protein (IUP). Analysis of the liquid fraction (bio-oil) obtained from the pyrolysis of microalgae Nannochloropsis gaditana, Scenedesmus spp with a bench scale reactor showed that DKPs constituted an important pool of nitrogen-containing compounds. Conversely, the level of DKPs was rather low in the bio-oil of Botryococcus braunii. The developed micropyrolysis procedure was applied in combination with thermogravimetry (TGA) and infrared spectroscopy (FT-IR) to investigate surface interaction between BSA and synthetic chrysotile. The results showed that the thermal behavior of BSA (e.g. DKPs formation) was affected by the different form of doped synthetic chrysotile. The typical DKPs evolved from collagen were quantified in the pyrolysates of archaeological bones from Vicenne Necropolis in order to evaluate their conservation status in combination with TGA, FTIR and XRD analysis.

Melanocytes: a new potential tool to study and monitoring Duchenne muscular dystrophy

Dystrophin is a subsarcolemmal protein critical for the integrity of muscle fibers by linking the actin cytoskeleton to the extracellular matrix via the dystroglycan complex. It is reported that dystroglycans are also localized in the skin, at dermal-epidermal junction. Here we show that epidermal melanocytes express dystrophin at the interface with the basement membrane. The full-length muscle isoform mDp427 was clearly detectable in epidermis and in melanocyte cultures as assessed by RNA and western blot analysis. Dystrophin was absent in Duchenne Muscular Dystrophy (DMD) patients melanocytes, and the ultrastructural analysis revealed mitochondrial alterations, similar to those occurring in myoblasts from the same patients. Interestingly, mitochondrial dysfunction of DMD melanocytes reflected the alterations identified in dystrophin-deficient muscle cells. In fact, mitochondria of melanocytes from DMD patients accumulated tetramethylrhodamine methyl ester but, on the contrary of control donor, mitochondria of DMD patients readily depolarized upon the addition of oligomycin, suggesting either that they are maintaining the membrane potential at the expense of glycolytic ATP, or that they are affected by a latent dysfunction unmasked by inhibition of the ATP synthase. Melanocyte cultures can be easily obtained by conventional skin biopsies, less invasive procedure than muscular biopsy, so that they may represent an alternative cellular model to myoblast for studying and monitoring dystrophinopathies also in response to pharmacological treatments.

Stratigraphic study and analysis of the barchan dunefield of the Laguna coastal zone in Santa Catarina, Brazil

The association of several favorable factors has resulted in the development of a wide barchan dune field that stands out as a fundamental element in the coastal landscape of southern Santa Catarina state in Brazil. This original ecosystem is being destroyed and highly modified, due to urbanization. This work identifies and discusses its basic characteristics and analyzes the favorable factors for its preservation, in the foreseen of both a sustainable future and potential incomes from ecotourism. The knowledge of the geologic evolution allows to associate this transgressive Holocene dunes formation to more dissipative beach conditions. Spatial differences on morphodynamics are related to local and regional contrasts in the sediment budget, with an influence on gradients of wave attenuation in the inner shelf and consequently with influence in the level of coastal erosion. The link between relative sea level changes and coastal eolian sedimentation can be used to integrate coastal eolian systems to the sequence stratigraphy model. The main accumulation phase of eolian sediments would occur during the final transgressive and highstand systems tracts. Considering the global character of Quaternary relative sea level changes, the Laguna transgressive dune field should be correlated with similar eolian deposits developed along other parts of the Brazilian coast compatibles with the model of dunefield initiation during rising and highstand sea level phases.

Dynamic computer simulations of electrophoresis: a versatile research and teaching tool

Software is available, which simulates all basic electrophoretic systems, including moving boundary electrophoresis, zone electrophoresis, ITP, IEF and EKC, and their combinations under almost exactly the same conditions used in the laboratory. These dynamic models are based upon equations derived from the transport concepts such as electromigration, diffusion, electroosmosis and imposed hydrodynamic buffer flow that are applied to user-specified initial distributions of analytes and electrolytes. They are able to predict the evolution of electrolyte systems together with associated properties such as pH and conductivity profiles and are as such the most versatile tool to explore the fundamentals of electrokinetic separations and analyses. In addition to revealing the detailed mechanisms of fundamental phenomena that occur in electrophoretic separations, dynamic simulations are useful for educational purposes. This review includes a list of current high-resolution simulators, information on how a simulation is performed, simulation examples for zone electrophoresis, ITP, IEF and EKC and a comprehensive discussion of the applications and achievements.

Borrelia in granuloma annulare, morphea and lichen sclerosus: a PCR-based study and review of the literature

Morphea, granuloma annulare (GA) and lichen sclerosus et atrophicans (LSA) have also been suggested to be linked to Borrelia infection. Previous studies based on serologic data or detection of Borrelia by immunohistochemistry and polymerase chain reaction (PCR) reported contradictory results. Thus, we examined skin biopsies of morphea, GA and LSA by PCR to assess the prevalence of Borrelia DNA in an endemic area and to compare our results with data in the literature.

Can S-100B serum protein help to save cranial CT resources in a peripheral trauma centre? A study and consensus paper

Cranial CT (CCT) is the gold standard to rule out traumatic brain injury. The serum level of the protein S-100B has recently been proposed as promising marker of traumatic brain injury. We prospectively investigated whether it might be a reliable tool for CCT triage in mild brain injury at a peripheral trauma centre with limited CT resources.

A comparison of initial antiretroviral therapy in the Swiss HIV Cohort Study and the recommendations of the International AIDS Society-USA

In order to facilitate and improve the use of antiretroviral therapy (ART), international recommendations are released and updated regularly. We aimed to study if adherence to the recommendations is associated with better treatment outcomes in the Swiss HIV Cohort Study (SHCS).

The Pararectus approach for anterior intrapelvic management of acetabular fractures: an anatomical study and clinical evaluation

A new anterior intrapelvic approach for the surgical management of displaced acetabular fractures involving predominantly the anterior column and the quadrilateral plate is described. In order to establish five 'windows' for instrumentation, the extraperitoneal space is entered along the lateral border of the rectus abdominis muscle. This is the so-called 'Pararectus' approach. The feasibility of safe dissection and optimal instrumentation of the pelvis was assessed in five cadavers (ten hemipelves) before implementation in a series of 20 patients with a mean age of 59 years (17 to 90), of whom 17 were male. The clinical evaluation was undertaken between December 2009 and December 2010. The quality of reduction was assessed with post-operative CT scans and the occurrence of intra-operative complications was noted. In cadavers, sufficient extraperitoneal access and safe instrumentation of the pelvis were accomplished. In the patients, there was a statistically significant improvement in the reduction of the fracture (pre- versus post-operative: mean step-off 3.3 mm (sd 2.6) vs 0.1 mm (sd 0.3), p < 0.001; and mean gap 11.5 mm (sd 6.5) vs 0.8 mm (sd 1.3), p < 0.001). Lesions to the peritoneum were noted in two patients and minor vascular damage was noted in a further two patients. Multi-directional screw placement and various plate configurations were feasible in cadavers without significant retraction of soft tissues. In the treatment of acetabular fractures predominantly involving the anterior column and the quadrilateral plate, the Pararectus approach allowed anatomical restoration with minimal morbidity related to the surgical access.

Genetic diversity of EBV-encoded LMP1 in the Swiss HIV Cohort Study and implication for NF-Κb activation

Epstein-Barr virus (EBV) is associated with several types of cancers including Hodgkin's lymphoma (HL) and nasopharyngeal carcinoma (NPC). EBV-encoded latent membrane protein 1 (LMP1), a multifunctional oncoprotein, is a powerful activator of the transcription factor NF-κB, a property that is essential for EBV-transformed lymphoblastoid cell survival. Previous studies reported LMP1 sequence variations and induction of higher NF-κB activation levels compared to the prototype B95-8 LMP1 by some variants. Here we used biopsies of EBV-associated cancers and blood of individuals included in the Swiss HIV Cohort Study (SHCS) to analyze LMP1 genetic diversity and impact of sequence variations on LMP1-mediated NF-κB activation potential. We found that a number of variants mediate higher NF-κB activation levels when compared to B95-8 LMP1 and mapped three single polymorphisms responsible for this phenotype: F106Y, I124V and F144I. F106Y was present in all LMP1 isolated in this study and its effect was variant dependent, suggesting that it was modulated by other polymorphisms. The two polymorphisms I124V and F144I were present in distinct phylogenetic groups and were linked with other specific polymorphisms nearby, I152L and D150A/L151I, respectively. The two sets of polymorphisms, I124V/I152L and F144I/D150A/L151I, which were markers of increased NF-κB activation in vitro, were not associated with EBV-associated HL in the SHCS. Taken together these results highlighted the importance of single polymorphisms for the modulation of LMP1 signaling activity and demonstrated that several groups of LMP1 variants, through distinct mutational paths, mediated enhanced NF-κB activation levels compared to B95-8 LMP1.

A Study and Application of the Process of Supergene Enrichment of Silver Ores.

In many deposits of silver ores the grade of the ore de­creases considerably a few hundred feet below the surface. It is believed that in many cases the better ores owe their richness in part to the process of sulphide enrichment. It is recognized, however, that many rich silver ores are hypogene deposits that have been affected very little, if any, by processes of enrichment.

Study and Correlation of Belt and Cambrian Arkoses near Limespur, Montana

In the vicinity of Limespur, Montana, a siding along the Northern Pacific Railroad near Whitehall, Montana, occurs a characteristic type of arkose where­ in many small red mineral grains are distributed throughout the rock mass. It is in this respect that this arkose differs from other arkoses in the surrounding region.