Chemical probes of surface layer biogenesis in Clostridium difficile

The bacterium Clostridium difficile is responsible for recent epidemics of gastroenteritis and currently causes over twice as many deaths per year as the other major hospital ‘superbug’, MRSA. Since the bacterium is resistant to conventional antibiotics there is an urgent need to develop novel therapies. C. difficile secretes a family of proteins that are held in place on the cell wall by non-covalent forces, producing a proteinaceous coat (the surface layer or S-layer) that surrounds the entire cell. These S-layer proteins (SLPs) are immunogenic in humans and play a role in binding to host cells. Synthesis of the C. difficile S-layer involves site-specific proteolytic cleavage of the SlpA precursor by an as yet unidentified C. difficile protease. Identification of the protease that processes SlpA has proven challenging, due in part to a lack of established genetic tools in C. difficle. Here the development of novel chemical probes that can disrupt S-layer formation by inhibiting the protease are described, and found to be powerful chemical proteomic tools for both protease identification and exploring the process of S-layer formation. Screening and inhibition experiments were first performed to identify novel synthetic irreversible protease inhibitors combining an electrophilic warhead with a specific sequence element matching the SlpA cleavage. These compound series were shown to possess structure-dependent activity, and inhibited cultures were also more sensitive to lysozyme-induced cell lysis, suggesting that correct processing and assembly of the S-layer is important for cell envelope integrity. Optimised inhibitors were further developed into ‘activity-based probes’ (ABPs) carrying an affinity tag, and were successfully used to isolate and identify de novo the key protease involved in cleavage of SlpA, Cwp84, using a combination of different labelling and proteomics approaches. These probes also permitted identification of Cwp84 activity across a wide range of clinical strains. In later work, the scope of the warhead element was explored in more detail, and the potential antibacterial effects of the inhibitors were investigated in both wild type and genetically engineered strains. Finally, the contributions of this work in terms of both chemical technology and C. difficile biology are critically assessed

Testing for Food market integration: A study of the Vietnamese paddy market

With its increasing integration into the world economy, agricultural exports and rural incomes in Vietnam have increased substantially in recent years. At the sub-national level, however, there are concerns that not all regions and categories of agricultural producers have and will benefit from the ongoing liberalization of agricultural markets. Vietnam's elongated geography and lack of spatial market integration pose special problems in this regard. Accordingly, this study aims to answer three interrelated questions: (a) whether there is spatial integration between paddy markets in the North and South of Vietnam; (b) whether there is spatial integration in paddy markets within the North and within the South; and, (c) if within-region integration is stronger and faster than between-region integration. The empirical model we develop to answer these questions, uses estimates of transfer costs to generalize the well known model of spatial market integration due to Ravallion to allow for the possibility of threshold effects. A sequential testing strategy is developed which progressively tests for market segmentation, the number of thresholds, long-run market integration, common dynamics/informational efficiency, and (a strict version of) the 'Law' of One Price within an error-correction framework. When the unrestricted version of this model is estimated using monthly paddy prices for eight markets between 1993 and 2006, we find weak evidence of market integration between paddy markets in the North and South of Vietnam with an absence of threshold effects. However, there is evidence of both threshold effects and stronger forms of spatial market integration for paddy markets within the North and within the South, with at least 60% percent of price changes being transmitted between markets within one month whenever price spreads exceeds their upper or lower thresholds. The extent and speed of price transmission within regional paddy markets is generally faster in the South than the North of Vietnam. However, the instantaneous version of the 'Law' of One Price, which requires full price adjustment to occur within a month, only holds for a few regimes and market pairs. Three main policy implications flow from these results. First, since there is limited evidence of integration between paddy markets in the North and South of Vietnam, national level policies cannot be relied upon to stabilize or support paddy prices.Second, since there is evidence of spatial market integration within the Red River and Mekong River deltas, paddy markets within these regions can be relied upon to transmit price signals between deficit and surplus areas relatively well. Third, since the speed and extent of price transmission is relatively rapid within the North and within the South of Vietnam, the private sector trade can be relied upon to transfer rice and paddy between markets in an efficient manner. Problems might, however, emerge if large demand-supply imbalances were to emerge between the North and South, as transfer costs would prevent private sector trade taking place. In these circumstances, the public sector might need to intervene, in a consistent and market friendly way, to ensure adequate food supplies in the short-term.Market integration; Paddy market; Error-correction; Spatial integration; Vietnam

Leases

In early 2016, the Financial Accounting Standards Board issued updated leasing standards intended to improve the financial reporting of leases. The new standards require lessees to report leases with terms of more than 12 months to be on the balance sheet. The new FASB standards is anticipated to provide a more faithful representation regarding leases to investors and financial statement users. In this study, I will provide a brief discussion about the new leasing standards. Second, I will explore the role technology and information systems plays in overcoming the difficulties with adopting the new leasing standards. Lastly will be an exploration concerning the costs of implementing this new standard

Extracting Fetal Electrocardiogram from Being Pregnancy Based on Nonlinear Projection

Fetal heart rate extraction from the abdominal ECG is of great importance due to the information that carries in assessing appropriately the fetus well-being during pregnancy. In this paper, we describe a method to suppress the maternal signal and noise contamination to discover the fetal signal in a single-lead fetal ECG recordings. We use a locally linear phase space projection technique which has been used for noise reduction in deterministically chaotic signals. Henceforth, this method is capable of extracting fetal signal even when noise and fetal component are of comparable amplitude. The result is much better if the noise is much smaller (P wave and T wave can be discovered)

Chronic scrotal heat stress causes testicular interstitial inflammation and fibrosis: An experimental study in mice

Background: Chronic heat stress is a risk factor that adversely affects the reproduction system. Inflammation and fibrosis are 2 important response processes to damaged tissues. Objective: This study investigates the association of chronic scrotal heat stress with testicular interstitial inflammation and fibrosis in mice. Materials and Methods: For all experiments, 8-10 wk old male Swiss mice (Mus musculus) (20-23 gr) were divided into 3 groups (n = 10/each). The heat-stress groups were submerged in a water bath at 37°C and 40°C, while the control group was treated at 25°C. The testicular tissues underwent hematoxylin and eosin staining, picro sirius red staining, and immunohistochemistry for intercellular adhesion molecule-1, fibroblast-specific protein 1, F4/80, collagen I, and Ki-67 staining to determine the testicular interstitial inflammation and fibrosis. Results: Chronic scrotal heat stress impairs spermatogenesis and reverses testicular histological structure. In this study, heat stress significantly induced increased interstitial cell proliferation and upregulation of intercellular adhesion molecule-1 expression in the interstitial testicular tissue. In the interstitial testicular tissue, the number of F4/80-positive macrophages and the number of fibroblast-specific protein 1- positive fibroblasts were significantly increased in the heat-exposed groups compared to those in the control group. The heat exposed groups had substantially increased extracellular matrix collagen accumulation in their testicular interstitial tissues. Conclusion: Heat stress adversely affects the testicular structure and spermatogenesis, causes inflammation, and leads to testicular interstitial fibrosis. Key words: Heat stress, Testicular, Inflammation, Fibrosis

Revisiting conventional noncovalent interactions towards a complete understanding: from tetrel to pnicogen, chalcogen, and halogen bond

Typical noncovalent interactions, including tetrel (TtB), pnicogen (PniB), chalcogen (ChalB), and halogen bonds (HalB), were systematically re-investigated by modeling the N⋯Z interactions (Z = Si, P, S, Cl) between NH$_3$ – as a nucleophilic, and SiF$_4$, PF$_3$, SF$_2$, and ClF – as electrophilic components, employing highly reliable ab initio methods. The characteristics of N⋯Z interactions when Z goes from Si to Cl, were examined through their changes in stability, vibrational spectroscopy, electron density, and natural orbital analyses. The binding energies of these complexes at CCSD(T)/CBS indicate that NH$_3$ tends to hold tightly most with ClF (−34.7 kJ mol$^{−1}$) and SiF$_4$ (−23.7 kJ mol$^{−1}$) to form N⋯Cl HalB and N⋯Si TtB, respectively. Remarkably, the interaction energies obtained from various approaches imply that the strength of these noncovalent interactions follows the order: N⋯Si TtB > N⋯Cl HalB > N⋯S ChalB > N⋯P PniB, that differs the order of their corresponding complex stability. The conventional N⋯Z noncovalent interactions are characterized by the local vibrational frequencies of 351, 126, 167, and 261 cm$^{−1}$ for TtB, PniB, ChalB, and HalB, respectively. The SAPT2+(3)dMP2 calculations demonstrate that the primary force controlling their strength retains the electrostatic term. Accompanied by the stronger strength of N⋯Si TtB and N⋯Cl HalB, the AIM and NBO results state that they are partly covalent in nature with amounts of 18.57% and 27.53%, respectively. Among various analysis approaches, the force constant of the local N⋯Z stretching vibration is shown to be most accurate in describing the noncovalent interactions