Absorption and metabolism of olive oil secoiridoids in the small intestine

The secoiridoids 3,4-dihydroxyphenylethanol-elenolic acid (3,4-DHPEA-EA) and 3,4-dihydroxyphenylethanol-elenolic acid dialdehyde (3,4-DHPEA-EDA) account for approximately 55 % of the phenolic content of olive oil and may be partly responsible for its reported human health benefits. We have investigated the absorption and metabolism of these secoiridoids in the upper gastrointestinal tract. Both 3,4-DHPEA-EDA and 3,4-DHPEA-EA were relatively stable under gastric conditions, only undergoing limited hydrolysis. Both secoiridoids were transferred across a human cellular model of the small intestine (Caco-2 cells). However, no glucuronide conjugation was observed for either secoiridoid during transfer, although some hydroxytyrosol and homovanillic alcohol were formed. As Caco-2 cells are known to express only limited metabolic activity, we also investigated the absorption and metabolism of secoiridoids in isolated, perfused segments of the jejunum and ileum. Here, both secoiridoids underwent extensive metabolism, most notably a two-electron reduction and glucuronidation during the transfer across both the ileum and jejunum. Unlike Caco-2 cells, the intact small-intestinal segments contain NADPH-dependent aldo-keto reductases, which reduce the aldehyde carbonyl group of 3,4-DHPEA-EA and one of the two aldeydic carbonyl groups present on 3,4-DHPEA-EDA. These reduced forms are then glucuronidated and represent the major in vivo small-intestinal metabolites of the secoiridoids. In agreement with the cell studies, perfusion of the jejunum and ileum also yielded hydroxytyrosol and homovanillic alcohol and their respective glucuronides. We suggest that the reduced and glucuronidated forms represent novel physiological metabolites of the secoiridoids that should be pursued in vivo and investigated for their biological activity.

Molecular modelling studies of binding of DACD derivatives into G-Quadruplex DNA: comparison of force field and quantum polarized ligand docking methods

The DNA G-qadruplexes are one of the targets being actively explored for anti-cancer therapy by inhibiting them through small molecules. This computational study was conducted to predict the binding strengths and orientations of a set of novel dimethyl-amino-ethyl-acridine (DACA) analogues that are designed and synthesized in our laboratory, but did not diffract in Synchrotron light.Thecrystal structure of DNA G-Quadruplex(TGGGGT)4(PDB: 1O0K) was used as target for their binding properties in our studies.We used both the force field (FF) and QM/MM derived atomic charge schemes simultaneously for comparing the predictions of drug binding modes and their energetics. This study evaluates the comparative performance of fixed point charge based Glide XP docking and the quantum polarized ligand docking schemes. These results will provide insights on the effects of including or ignoring the drug-receptor interfacial polarization events in molecular docking simulations, which in turn, will aid the rational selection of computational methods at different levels of theory in future drug design programs. Plenty of molecular modelling tools and methods currently exist for modelling drug-receptor or protein-protein, or DNA-protein interactionssat different levels of complexities.Yet, the capasity of such tools to describevarious physico-chemical propertiesmore accuratelyis the next step ahead in currentresearch.Especially, the usage of most accurate methods in quantum mechanics(QM) is severely restricted by theirtedious nature. Though the usage of massively parallel super computing environments resulted in a tremendous improvement in molecular mechanics (MM) calculations like molecular dynamics,they are still capable of dealing with only a couple of tens to hundreds of atoms for QM methods. One such efficient strategy that utilizes thepowers of both MM and QM are the QM/MM hybrid methods. Lately, attempts have been directed towards the goal of deploying several different QM methods for betterment of force field based simulations, but with practical restrictions in place. One of such methods utilizes the inclusion of charge polarization events at the drug-receptor interface, that is not explicitly present in the MM FF.

Eliminating the small-angle component of the scattering calculated for models

A straightforward procedure (assuming spherical symmetry) is described, which enables the unwanted small-angle component of the scattering for a finite model to be calculated. The method may be applied to models of any shape or size. It is illustrated by means of a single polymer chain.

Poverty traps in small-scale mining communities: the case of sub-Saharan Africa

This paper critically reflects on why, in many rural stretches of sub-Saharan Africa, scores of people engage in artisanal and small-scale mining (ASM) activity – low-tech, labour intensive mineral extraction – for lengthy periods of time. It argues that a large share of the region’s ASM operators have mounting debts which prevent them from pursuing alternative, less arduous, employment. The paper concludes with an analysis of findings from research carried out by the author in Talensi-Nabdam District, Northern Ghana, which captures the essence of the poverty trap now plaguing so many ASM communities in sub-Saharan Africa.

Chemical characterisation and determination of sensory attributes of hydrolysates produced by enzymatic hydrolysis of whey proteins following a novel integrative process

The overall aim of this work was to characterize the major angiotensin converting enzyme (ACE) inhibitory peptides produced by enzymatic hydrolysis of whey proteins, through the application of a novel integrative process. This process consisted of the combination of adsorption and microfiltration within a stirred cell unit for the selective immobilization of β-lactoglobulin and casein derived peptides (CDP) from whey. The adsorbed proteins were hydrolyzed in-situ which resulted in the separation of peptide products from the substrate and fractionation of peptides. Two different hydrolysates were produced: (i) from CDP (IC50 =287μg/mL) and (ii) from β-lactoglobulin (IC50=128μg/mL). IC50 is the concentration of inhibitor needed to inhibit ACE by half. The well known antihypertensive peptide IPP and several novel peptides that have structural similarities with reported ACE inhibitory peptides were identified and characterized in both hydrolysates. Furthermore, the hydrolysates were assessed for bitterness. No significant difference was found between the control (milk with no hydrolysate) and hydrolysate samples at different concentrations (at, below and above the IC50).

Large is fast, small is tight: determinants of speed and affinity in subunit capture by a periplasmic chaperone

The chaperone/usher pathway assembles surface virulence organelles of Gram-negative bacteria, consisting of fibers of linearly polymerized protein subunits. Fiber subunits are connected through 'donor strand complementation': each subunit completes the immunoglobulin (Ig)-like fold of the neighboring subunit by donating the seventh β-strand in trans. Whereas the folding of Ig domains is a fast first-order process, folding of Ig modules into the fiber conformation is a slow second-order process. Periplasmic chaperones separate this process in two parts by forming transient complexes with subunits. Interactions between chaperones and subunits are also based on the principle of donor strand complementation. In this study, we have performed mutagenesis of the binding motifs of the Caf1M chaperone and Caf1 capsular subunit from Yersinia pestis and analyzed the effect of the mutations on the structure, stability, and kinetics of Caf1M-Caf1 and Caf1-Caf1 interactions. The results suggest that a large hydrophobic effect combined with extensive main-chain hydrogen bonding enables Caf1M to rapidly bind an early folding intermediate of Caf1 and direct its partial folding. The switch from the Caf1M-Caf1 contact to the less hydrophobic, but considerably tighter and less dynamic Caf1-Caf1 contact occurs via the zip-out-zip-in donor strand exchange pathway with pocket 5 acting as the initiation site. Based on these findings, Caf1M was engineered to bind Caf1 faster, tighter, or both faster and tighter. To our knowledge, this is the first successful attempt to rationally design an assembly chaperone with improved chaperone function.

Changes in area and geodetic mass balance of small glaciers, polar Urals, Russia, 1950-2008

Changes in area of 30 small glaciers (mostly <1 km2) in the northern Polar Urals (67.5-68.25 °N) between 1953 and 2000 were assessed using historic aerial photography from 1953 and 1960, ASTER and panchromatic Landsat ETM+ imagery from 2000, and data from 1981 and 2008 terrestrial surveys. Changes in volume and geodetic mass balance of IGAN and Obruchev glaciers were calculated using data from terrestrial surveys in 1963 and 2008. In total, glacier area declined by 22.3 ± 3.9% in the 1953/60-2000 period. The areas of individual glaciers decreased by 4-46%. Surfaces of Obruchev and IGAN glaciers lowered by 22.5 ± 1.7 m and 14.9 ± 2.1 m. Over 45 years, geodetic mass balances of Obruchev and IGAN glaciers were -20.66 ± 2.91 and -13.54 ± 2.57 m w.e. respectively. Glacier shrinkage in the Polar Urals is related to a summer warming of 1 °C between 1953-81 and 1981-2008 and its rates are consistent with other regions of northern Asia but are higher than in Scandinavia. While glacier shrinkage intensified in the 1981-2000 period relative to 1953-81, increasing winter precipitation and shading effects slowed glacier wastage in 2000-08.

Rhinocetin, a venom-derived integrin-specific antagonist inhibits collagen-induced platelet and endothelial cell functions

Snaclecs are small non-enzymatic proteins present in viper venoms reported to modulate haemostasis of victims through effects on platelets, vascular endothelial and smooth muscle cells. In this study, we have isolated and functionally characterised a snaclec which we named rhinocetin from the venom of West African gaboon viper, Bitis gabonica rhinoceros. Rhinocetin was shown to comprise α and β chains with the molecular masses of 13.5 and 13kDa respectively. Sequence and immunoblot analysis of rhinocetin confirmed this to be a novel snaclec. Rhinocetin inhibited collagen-stimulated activation of human platelets in dose dependent manner, but displayed no inhibitory effects on glycoprotein VI (collagen receptor) selective agonist, CRP-XL-, ADP- or thrombin-induced platelet activation. Rhinocetin antagonised the binding of monoclonal antibodies against the α2 subunit of integrin α2β1 to platelets and coimmunoprecipitation analysis confirmed integrin α2β1 as a target for this venom protein. Rhinocetin inhibited a range of collagen induced platelet functions such as fibrinogen binding, calcium mobilisation, granule secretion, aggregation and thrombus formation. It also inhibited integrin α2β1 dependent functions of human endothelial cells. Together, our data suggest rhinocetin to be a modulator of integrin α2β1 function and thus may provide valuable insights into the role of this integrin in physiological and pathophysiological scenarios including haemostasis, thrombosis and envenomation.

Good practice transfer within small construction specialist trade contractors

Government and institutionally-driven ‘good practice transfer’ initiatives are consistently presented as a means to enhance construction firm and industry performance. Two implicit tenets of these initiatives appear to be: knowledge embedded in good practice will transfer automatically; and, the potential of implementing good practice will be capitalised regardless of the context where it is to be used. The validity of these tenets is increasingly being questioned and, concurrently, more nuanced knowledge production understandings are being developed which recognise and incorporate context-specificity. This research contributes to this growing, more critical agenda by examining the actual benefits accrued from good practice transfer from the perspective of a small specialist trade contracting firm. A concept model for successful good practice transfer is developed from a single longitudinal case study within a small heating and plumbing firm. The concept model consists of five key variables: environment, strategy, people, technology, and organisation of work. The key findings challenge the implicit assumptions prevailing in the existing literature and support a contingency approach that argues successful good practice transfer is not just adopting and mechanistically inserting into the firm, but requires addressing ‘behavioural’ aspects. For successful good practice transfer, small specialist trade contracting firms need to develop and operationalise organisation slack, mechanisms for scanning external stimuli and absorbing knowledge. They also need to formulate and communicate client-driven external strategies; to motive and educate people at all levels; to possess internal or accessible complementary skills and knowledge; to have ‘soft focus’ immediate/mid-term benefits at a project level; and, to embed good practice in current work practices.

Sleepwalking certainties: agency, aesthetics, and incapacity in W.G. Sebald’s 'Austerlitz' and Hermann Broch’s 'The Sleepwalkers'

When we first encounter the narrator of Austerlitz, he is wandering around the unfamiliar town of Antwerp with, he tells us, “unsicheren Schritten” (1; 9). As well as reflecting the unfamiliarity of the locale, these “uncertain steps” evince a proud modesty characteristic of the classic Sebaldian narrator, a wanderer who discreetly relays the stories of the people and places he is privileged to encounter. Although Sebald does not use the phrase, steps of this sort, unpurposed yet unerring, are made with what is commonly known in German as somnambule Sicherheit: the legendary surefootedness of the sleepwalker. The convergence of sleepwalking and certainty in a single phrase poses an interesting challenge to one of the central tenets of the English-language canonization of Sebald, for his writing has been most highly valued for its ability to move the reader through apparent certainties towards a salutary uncertainty. But somnambule Sicherheit also presents the possibility that the current may be reversed, that narrative may move under cover of uncertainty towards certainty. That Sebald criticism has not been more troubled by this possibility is in no small part due to the fact that it tends to deploy the notion of sleepwalking with a minimum of reflection on its theoretical ramifications. To evoke some of the complexities of this matter, I first offer a brief cultural history of sleepwalking, as well as a brief account of the topic of uncertainty in Sebald criticism. Most of my argument, however, involves an extended comparative analysis of sleepwalking in Sebald's Austerlitz and Hermann Broch's 1933 trilogy The Sleepwalkers. Although these writers have not previously been the object of any sustained comparison, sleepwalking in Broch's novels illuminates much that is left implicit on the topic in Sebald's fiction and points toward some difficult questions regarding the role of aesthetics and agency in Sebald's work.

Piezotolerant small-colony variants with increased thermotolerance, antibiotic susceptibility, and low invasiveness in a clonal Staphylococcus aureus population

Following a pressure treatment of a clonal Staphylococcus aureus culture with 400 MPa for 30 min, piezotolerant variants were isolated. Among 21 randomly selected survivors, 9 were piezotolerant and all formed small colonies on several agar media. The majority of the isolates showed increased thermotolerance, impaired growth, and reduced antibiotic resistance compared to the wild type. However, several nonpiezotolerant isolates also demonstrated impaired growth and the small-colony phenotype. In agglutination tests for the detection of protein A and fibrinogen, the piezotolerant variants showed weaker agglutination reactions than the wild type and the other isolates. All variants also showed defective production of the typical S. aureus golden color, a characteristic which has previously been linked with virulence. They were also less able to invade intestinal epithelial cells than the wild type. These S. aureus variants showed phenotypic similarities to previously isolated Listeria monocytogenes piezotolerant mutants that contained mutations in ctsR. Because of these similarities, possible alterations in the ctsR hypermutable regions of the S. aureus variants were investigated through amplified fragment length polymorphism analysis. No mutations were identified, and subsequently we sequenced the ctsR and hrcA genes of three representative variants, finding no mutations. This work demonstrates that S. aureus probably possesses a strategy resulting in an abundance of multiple-stressresistant variants within clonal populations. This strategy, however, seems to involve genes and regulatory mechanisms different from those previously reported for L. monocytogenes. We are in the process of identifying these mechanisms.