Mechanisms of Mycoplasma-induced inflammation and cellular transformation

There is a growing interest in “medical gasses” for their antibacterial and anti-inflammatory properties. Hydrogen sulfide (H2S), a member of the family of gasotransmitters, is in fact increasingly being recognized as an important signaling molecule, but its precise role in the regulation of the inflammatory response is still not clear. For this reason, the aim of the first part of this thesis was to investigate the effects of H2S on the expression of pro-inflammatory cytokines, such as MCP-1, by using an in vitro model composed by both primary monocytes-derived macrophages cultures and the human monocytic cell line U937 infected with Mycoplasma fermentans, a well-known pro-inflammatory agent. In our experiments, we observed a marked increase in the production of pro-inflammatory cytokines in infected cells. In particular, MCP-1 was induced both at the RNA and at the protein level. To test the effects of H2S on infected cells, we treated the cells with two different H2S donors (NaHS and GYY4137), showing that both H2S treatments had anti-inflammatory effects in Mycoplasma-infected cells: the levels of MCP-1, both mRNA expression and protein production, were reduced. Our subsequent studies aimed at understanding the molecular mechanisms responsible for these effects, focused on two specific molecular pathways, both involved in inflammation: the NF-κB and the Nrf2 pathway. After treatment with pharmacological inhibitors, we demonstrated that Mycoplasma fermentans induces MCP-1 expression through the TLR-NF-κB pathway with the nuclear translocation of its subunits, while treatment with H2S completely blocked the nuclear translocation of NF-κB heterodimer p65/p50. Then, once infected cells were treated with H2S donors, we observed an increased protective effect of Nrf2 and also a decrease in ROS production. These results highlight the importance of H2S in reducing the inflammatory process caused by Mycoplasma fermentans. To this regard, it should be noted that several projects are currently ongoing to develop H2S-releasing compounds as candidate drugs capable of alleviating cell deterioration and to reduce the rate of decline in organ function. In the second part of this study, we investigated the role of Mycoplasma infection in cellular transformation. Infectious agents are involved in the etiology of many different cancers and a number of studies are still investigating the role of microbiota in tumor development. Mycoplasma has been associated with some human cancers, such as prostate cancer and non-Hodgkin’s lymphoma in HIV-seropositive people, and its potential causative role and molecular mechanisms involved are being actively investigated. To this regard, in vitro studies demonstrated that, upon infection, Mycoplasma suppresses the transcriptional activity of p53, key protein in the cancer suppression. As a consequence, infected cells were less susceptible to apoptosis and proliferated more than the uninfected cells. The mechanism(s) responsible for the Mycoplasma-induced inhibitory effect on p53 were not determined. Aim of the second part of this thesis was to better understand the tumorigenic role of the microorganism, by investigating more in details the effect(s) of Mycoplasma on p53 activity in an adenocarcinoma HCT116 cell line. Treatment of Mycoplasma-infected cells with 5FU or with Nutlin, two molecules that induce p53 activity, resulted in cellular proliferation comparable to untreated controls. These results suggested that Mycoplasma infection inhibited p53 activity. Immunoprecipitation of p53 with specific antibodies, and subsequent Gas Chromatography and Mass Spectroscopy (GC-MS) assays, allowed us to identify several Mycoplasma-specific proteins interacting with p53, such as DnaK, a prokaryotic heat shock protein and stress inducible chaperones. In cells transfected with DnaK we observed i) reduced p53 protein levels; ii) reduced activity and expression of p21, Bax and PUMA, iii) a marked increase in cells leaving G1 phase. Taken together, these data show an interaction between the human p53 and the Mycoplasma protein DnaK, with the consequent decreased p53 activity and decreased capability to respond to DNA damage and prevent cell proliferation. Our data indicate that Mycoplasma could be involved in cancer formation and the mechanism(s) has the potential to be a target for cancer diagnosis and treatment(s).

Character recognition systems using Fourier transformation in incoherent light

Many attempts have been made to overcome problems involved in character recognition which have resulted in the manufacture of character reading machines. An investigation into a new approach to character recognition is described. Features for recognition are Fourier coefficients. These are generated optically by convolving characters with periodic gratings. The development of hardware to enable automatic measurement of contrast and position of periodic shadows produced by the convolution is described. Fourier coefficients of character sets were measured, many of which are tabulated. Their analysis revealed that a few low frequency sampling points could be selected to recognise sets of numerals. Limited treatment is given to show the effect of type face variations on the values of coefficients which culminated in the location of six sampling frequencies used as features to recognise numerals in two type fonts. Finally, the construction of two character recognition machines is compared and contrasted. The first is a pilot plant based on a test bed optical Fourier analyser, while the second is a more streamlined machine d(3signed for high speed reading. Reasons to indicate that the latter machine would be the most suitable to adapt for industrial and commercial applications are discussed.

Building team-based working:a practical guide to organizational transformation

Building Team-based Working is designed for use by managers and consultants who are introducing team-based working into organizations. The book synthesizes knowledge about how to build team-based organizations, focusing particularly on the psychological and social processes that can facilitate or obstruct successful teamwork. Rather than advise managers on how to build effective teams, as most books in this area tend to do, this book instead focuses on how to build organizations structured around teams. The text is divided into six sections describing the six main stages of developing team-based working in an organization. The chapters follow a common structure. Each one opens with a summary of the aims and activities relevant to that stage and concludes with a selection of appropriate support materials and tools. These materials can also be downloaded from the CD accompanying the text. The advice given is based on evidence gathered by the authors over 20 years of practical management experience, research work in organizations, and consultancy across the public, manufacturing and service sectors.

Block co-polymerization by transformation reactions

The aim of this study was to use the transformation of anionic to metathesis polymerization to produce block co-polymers of styrene-b-pentenylene using WC16 /PStLi and WC16/PStLi/ AlEtC12 catalyst systems. Analysis of the products using SEC and 1H and 13C NMR spectroscopy enabled mechanisms for metathesis initiation reactions to be proposed. The initial work involved preparation of the constituent homo-polymers. Solutions of polystyryllithium in cyclohexane were prepared and diluted so that the [PStLi]o<2x10-3M. The dilution produced initial rapid decay of the active species, followed by slower spontaneous decay within a period of days. This was investigated using UV / visible spectrophotometry and the wavelength of maximum absorbance of the PStLi was found to change with the decay from an initial value of 328mn. to λmax of approximately 340nm. after 4-7 days. SEC analysis of solutions of polystyrene, using RI and UV / visible (set at 254nm.) detectors; showed the UV:RI peak area was constant for a range of polystyrene samples of different moleculor weight. Samples of polypentenylene were prepared and analysed using SEC. Unexpectedly the solutions showed an absorbance at 254nm. which had to be considered when this technique was used subsequently to analyse polymer samples to determine their styrene/ pentenylene co-polymer composition. Cyclohexane was found to be a poor solvent for these ring-opening metathesis polymerizations of cyclopentene. Attempts to produce styrene-b-pentenylene block co-polymers, using a range of co-catalyst systems, were generally unsuccessful as the products were shown to be mainly homopolymers. The character of the polymers did suggest that several catalytic species are present in these systems and mechanisms have been suggested for the formation of initiating carbenes. Evidence of some low molecular weight product with co-polymer character has been obtained. Further investigation indicated that this is most likely to be ABA block copolymer, which led to a mechanism being proposed for the termination of the polymerization.

Virtual manufacturing:a study of some important issues relating to the transformation of traditional manufacturing organisations

This thesis starts with a literature review, outlining the major issues identified in the literature concerning virtual manufacturing enterprise (VME) transformation. Then it details the research methodology used – a systematic approach for empirical research. next, based on the conceptual framework proposed, this thesis builds three modules to form a reference model, with the purpose of clarifying the important issues relevant to transforming a traditional manufacturing company into a VME. The first module proposes a mechanism of VME transformation – operating along the VME metabolism. The second module builds a management function within a VME to ensure a proper operation of the mechanism. This function helps identify six areas as closely related to VME transformation: lean manufacturing; competency protection; internal operation performance measurement; alliance performance measurement; knowledge management; alliance decision making. The third module continues and proposes an alliance performance measurement system which includes 14 categories of performance indicators. An analysis template for alliance decision making is also proposed and integrated into the first module. To validate these three modules, 7 manufacturing organisations (5 in China and 2 in the UK) were investigated, and these field case studies are analysed in this thesis. The evidence found in these organisations, together with the evidence collected from the literature, including both researcher views and literature case studies, provide support for triangulation evidence. In addition, this thesis identifies the strength and weakness patterns of the manufacturing companies within the theoretical niche of this research, and clarifies the relationships among some major research areas from the perspective of virtual manufacturing. Finally, the research findings are summarised, as well as their theoretical and practical implications. Research limitations and recommendations for future work conclude this thesis.

Study of neutral Fe(III) complexes of pyridoxal-N-substituted thiosemicarbazone with desolvation induced spin-state transformation above room temperature

The preparation and characterization of two new neutral ferric complexes with desolvation-induced discontinuous spin-state transformation above room temperature are reported. The compounds, Fe(Hthpy)(thpy).CH3OH.3H2O (1) and Fe(Hmthpy)(mthpy).2H2O (2), are low-spin (LS) at room temperature and below, whereas their nonsolvated forms are high-spin (HS), exhibiting zero-field splitting. In these complexes, Hthpy, Hmthpy, and thpy, mthpy are the deprotonated forms of pyridoxal thiosemicarbazone and pyridoxal methylthiosemicarbazone, respectively; each is an O,N,S-tridentate ligand. The molecular structures have been determined at 100(1) K using single-crystal X-ray diffraction techniques and resulted in a triclinic system (space group P1) and monoclinic unit cell (space group P21/c) for 1 and 2, respectively. Structures were refined to the final error indices, where RF = 0.0560 for 1 and RF = 0.0522 for 2. The chemical inequivalence of the ligands was clearly established, for the "extra" hydrogen atom on the monodeprotonated ligands (Hthpy, Hmthpy) was found to be bound to the nitrogen of the pyridine ring. The ligands are all of the thiol form; the doubly deprotonated chelates (thpy, mthpy) have C-S bond lengths slightly longer than those of the singly deprotonated forms. There is a three-dimensional network of hydrogen bonds in both compounds. The discontinuous spin-state transformation is accompanied with liberation of solvate molecules. This is evidenced also from DSC analysis. Heat capacity data for the LS and HS phases are tabulated at selected temperatures, the values of the enthalpy and entropy changes connected with the change of spin state were reckoned at DeltaH = 12.5 0.3 kJ mol-1 and DeltaS = 33.3 0.8 J mol-1 K-1, respectively, for 1 and DeltaH = 6.5 0.3 kJ mol-1 and DeltaS = 17.6 0.8 J mol-1 K-1, respectively, for 2

The verbal transformation effect:an exploration of the perceptual organization of speech

Six experiments investigated the influence of several grouping cues within the framework of the Verbal Transformation Effect (VTE, Experiments 1 to 4) and Phonemic Transformation Effect (PTE, Experiments 5 and 6), where listening to a repeated word (VTE) or sequence of vowels (PTE) produces verbal transformations (VTs). In Experiment 1, the influence of F0 frequency and lateralization cues (ITDs) was investigated in terms of the pattern of VTs. As the lateralization difference increased between two repeating sequences, the number of forms was significantly reduced with the fewest forms reported in the dichotic condition. Experiment 2 explored whether or not propensity to report more VTs on high pitch was due to the task demands of monitoring two sequences at once. The number of VTs reported was higher when listeners were asked to attend to one sequence only, suggesting smaller attentional constraints on the task requirements. In Experiment 3, consonant-vowel transitions were edited out from two sets of six stimuli words with ‘strong’ and ‘weak’ formant transitions, respectively. Listeners reported more forms in the spliced-out than in the unedited case for the strong-transition words, but not for those with weak transitions. A similar trend was observed for the F0 contour manipulation used in Experiment 4 where listeners reported more VTs and forms for words following a discontinuous F0 contour. In Experiments 5 and 6, the role of F0 frequency and ITD cues was investigated further using a related phenomenon – the PTE. Although these manipulations had relatively little effect on the number of VTs and forms reported, they did influence the particular forms heard. In summary, the current experiments confirmed that it is possible to successfully investigate auditory grouping cues within the VTE framework and that, in agreement with recent studies, the results can be attributed to the perceptual re-grouping of speech sounds.

Organizational change and development in India:a case of strategic organizational change and transformation

Purpose – The purpose of this paper is to examine developments in the field of organizational change (OC) with reference to the context of India. It highlights the need to analyze this topic in the present Indian economic environment and discusses the main developments reported in the Indian literature on the same. Design/methodology/approach – Empirical evidence based on a qualitative analysis of a case study undertaken at a public-private partnership transformation at North Delhi Power Limited (NDPL) in India is presented. Findings – The findings focus on trust building and belongingness for the employees, establishing a high-performance orientation, quality improvements, and the resultant transformations at NDPL. The analysis indicates a number of ways by which NDPL sought to improve its efficiency in order to better adapt to the rapidly changing Indian business environment. Practical implications – Based on the findings, the paper identifies key messages for policy makers and change agents regarding how to transform companies in the rapidly changing business contexts of emerging markets such as India. Originality/value – The paper offers an in-depth analysis of OC practices in a large organization in India.

Introduction:understanding the transformation of the CDU

This article introduces the special issue on the transformation of German Christian Democracy and sets the context for organisational and policy change in the CDU. As a starting point, it identifies both contemporary challenges to the CDU, locating its discussion within the wider literature on European Christian Democracy, and considers the extent to which these apply to the German case. It then outlines the key dimensions of the contemporary CDU which are examined in this collection. © 2013 Association for the Study of German Politics.