Search

This paper describes the interaction of the Crossrail project to deliver London's new Elizabeth line in the UK with a building situated directly over the machine-bored tunnels, close to Paddington station. The initial scheme called for the demolition of the existing building at 4–18 Bishop's Bridge Road, including extraction of existing piles, which would have had a considerable impact on the local area. Changes in the design of the Elizabeth line station at Paddington allowed a revised strategy to be considered, whereby the building could be kept, reducing construction costs to the project considerably. A combination of desk study and ground investigation works was used to identify the likely extent of the piling under the building, and to predict the risk of clashes with the tunnels. The results of analyses considering the impact of construction works on the building, and associated monitoring results are presented. The decision not to demolish produced numerous benefits aside from the reduction in cost, including a considerably reduced impact on those using the building and adjacent areas.

Main-text simulation result files Sarvari, P.; Ingram, D.; Stan, G.-B. A Modelling Framework Linking Resource-Based Stochastic Translation to the Optimal Design of Synthetic Constructs. Biology 2021, 10, 37. https://doi.org/10.3390/biology 10010037 Data ID is the number before the extension. No slow codon | Data ID| Prom_H | RBS_H| | :---------- | :------------: | ------------:| | 0 | 1/3 | 1/3 | | 1 | 1.0 | 1/3 | | 2 | 3.0 | 1/3 | | 3 | 1/3 | 1.0 | | 4 | 1.0 | 1.0 | | 5 | 3.0 | 1.0 | | 6 | 1/3 | 3.0 | | 7 | 1.0 | 3.0 | | 8 | 3.0 | 3.0 | Slow codon is 5th | Data ID| Prom_H | RBS_H| | :---------- | :------------: | ------------:| | 9 | 1/3 | 1/3 | | 10 | 1.0 | 1/3 | | 11 | 3.0 | 1/3 | | 12 | 1/3 | 1.0 | | 13 | 1.0 | 1.0 | | 14 | 3.0 | 1.0 | | 15 | 1/3 | 3.0 | | 16 | 1.0 | 3.0 | | 17 | 3.0 | 3.0 | Slow codon is 26th | Data ID| Prom_H | RBS_H| | :---------- | :------------: | ------------:| | 18 | 1/3 | 1/3 | | 19 | 1.0 | 1/3 | | 20 | 3.0 | 1/3 | | 21 | 1/3 | 1.0 | | 22 | 1.0 | 1.0 | | 23 | 3.0 | 1.0 | | 24 | 1/3 | 3.0 | | 25 | 1.0 | 3.0 | | 26 | 3.0 | 3.0 &nbsp

The protozoan pathogen Leishmania donovani encounters large fluctuations in its ionic and osmotic microenvironment as it cycles between the gut of the insect vector and the parasitophorous vacuole ...

As a result of the challenge of recent dust storms to public health, we tested the postulate that desert dust collected in the southwestern United States imparts a biological effect in respiratory epithelial cells and an animal model. Two samples of surface sediment were collected from separate dust sources in northeastern Arizona. Analysis of the PM20 fraction demonstrated that the majority of both dust samples were quartz and clay minerals (total SiO₂ of 52 and 57%). Using respiratory epithelial and monocytic cell lines, the two desert dusts increased oxidant generation, measured by Amplex Red fluorescence, along with carbon black (a control particle), silica, and NIST 1649 (an ambient air pollution particle). Cell oxidant generation was greatest following exposures to silica and the desert dusts. Similarly, changes in RNA for superoxide dismutase-1, heme oxygenase-1, and cyclooxygenase-2 were also greatest after silica and the desert dusts supporting an oxidative stress after cell exposure. Silica, desert dusts, and the ambient air pollution particle NIST 1649 demonstrated a capacity to activate the p38 and ERK1/2 pathways and release pro-inflammatory mediators. Mice, instilled with the same particles, showed the greatest lavage concentrations of pro-inflammatory mediators, neutrophils, and lung injury following silica and desert dusts. We conclude that, comparable to other particles, desert dusts have a capacity to (1) influence oxidative stress and release of pro-inflammatory mediators in respiratory epithelial cells and (2) provoke an inflammatory injury in the lower respiratory tract of an animal model. The biological effects of desert dusts approximated those of silica.

Wage claims have been an important feature of British industrial relations during the postwar period. They help set the boundaries within which wage negotiations take place and provide an insight into the conduct of negotiations, especially during periods of change in industrial relations. Despite this, claims remain an underinvestigated area. This article provides a unique investigation of the dimensions of wage claims over a period of free collective bargaining. The number of wage claims declined along with unionisation but, over a period of economic turbulence, the conduct of British wage setting began to change. We examine data on claims and investigate the influences on changes in those claims over time. We find that external factors (inflation, unemployment and legislative control of unions) were more prominent in shaping the development of claims than changes in the composition of groups who continued to post claims.

Pneumoconiosis is often considered a disease of the lung initiated by exposure to dust or other airborne particles, resulting in injury to the lungs. The term "endogenous pneumoconiosis" has been used in the literature to describe the deposition of compounds on the elastic fibers of the lung, usually in the setting of cardiac failure. In the case we present here, the patient aspirated a foreign body resulting in damage to the lung tissue and subsequent deposition of endogenous compounds on the elastic fibers of the pulmonary parenchyma and vasculature. We determined the composition of this mineral and mapped the distribution of elements using a combination of backscattered electron microscopy and energy dispersive spectrometry.

The election of the Conservative government in 1979 is seen as a watershed in pay determination in the UK, ending formal pay policies. While the government withdrew, the UK's major employer organisation, the Confederation of British Industry (CBI), endeavoured to operate an alternative system of employer coordination on pay between 1979 and 1997. Had this approach succeeded, the UK's approach to pay determination would have become much more aligned to the models adopted within other EU Member States. We analyse the development of this alternative to government-imposed incomes policy and measure its impact on the growth of earnings. We conclude that, despite the CBI's policy, the actions of individual employers suffered from classic market failure until the British economy was subjected to mounting competitive pressures from the international environment.

Michael Moon interviews Peter Ingram, Founder of BlendedBusiness, on marketing integration and automation within a marketing operations platform. This interview features a discussion on customer relationship management and how marketers and their agencies are looking for new ways to optimize customer engagement. In addition, Moon and Ingram review current challenges in supply chain management as well as the change management required within organizations to better orient themselves around retail marketing and shopper marketing.

The synthesis and antibacterial activities of three chemotypes of DNA supercoiling inhibitors based on imidazolo[1,2-a]pyridine and [1,2,4]triazolo[1,5-a]pyridine scaffolds that target the ATPase subunits of DNA gyrase and topoisomerase IV (GyrB/ParE) is reported. The most potent scaffold was selected for optimization leading to a series with potent Gram-positive antibacterial activity and a low resistance frequency.

The general trend of microscopical investigation in biology from the 1950s to the early 1970s was towards obtaining structural information. This goal initially was met using heavy metal and/or aldehyde fixatives, room temperature dehydration with polar organic liquids, embedding with epoxy and acrylate resins, and thin sectioning at room temperature. By the mid 1970s, a perceptible change occurred in the direction of both light and analytical electron microscopy towards investigation of the chemical reactivity and composition of structures made visible with increasingly better spatial resolution for light and electron microscopes. During the past 25 years there have been considerable innovations in microanalytical techniques, including analytical electron and x-ray microscopy and microanalysis, secondary ion mass spectrometry, laser microprobe mass analysis, the scanning probe microscopies, and confocal/multi-photon microscopy. In addition, cryopreservation and the development of chromophores for visualization of molecular and ionic sites within individual living cells as well as membranes have redefined the goal of microscopical preservation: to stabilize cell structure and composition as they exist in the living state.

Previous studies of the yeast Saccharomyces cerevisiae indicated that the vacuole is a major site of zinc storage in the cell. However, these studies did not address the absolute level of zinc that was stored in the vacuole nor did they examine the abundances of stored zinc in other compartments of the cell. In this report, we describe an analysis of the cellular distribution of zinc by use of both an organellar fractionation method and an electron probe X-ray microanalysis. With these methods, we determined that zinc levels in the vacuole vary with zinc status and can rise to almost 100 mM zinc (i.e., 7 × 10 8 atoms of vacuolar zinc per cell). Moreover, this zinc can be mobilized effectively to supply the needs of as many as eight generations of progeny cells under zinc starvation conditions. While the Zrc1 and Cot1 zinc transporters are essential for zinc uptake into the vacuole under steady-state growth conditions, additional transporters help mediate zinc uptake into the vacuole during “zinc shock,” when zinc-limited cells are resupplied with zinc. In addition, we found that other compartments of the cell do not provide significant stores of zinc. In particular, zinc accumulation in mitochondria is low and is homeostatically regulated independently of vacuolar zinc storage. Finally, we observed a strong correlation between zinc status and the levels of magnesium and phosphorus accumulated in cells. Our results implicate zinc as a major determinant of the ability of the cell to store these other important nutrients.

The protozoan parasite Leishmania donovani encounters large fluctuations in osmolality as it cycles between its insect vector and human host. The flagellated promastigote exhibits regulatory volume responses involving organic and inorganic osmolytes, but little is known about volume regulation in the clinically relevant amastigote that multiplies within the parasitophorous vacuoles of mammalian host cells. Using a combination of morphological, X-ray microanalytical, and biochemical approaches we determined that non-motile amastigotes respond to hypotonic stress with (1) an amino acid and l-alanine-mediated regulatory volume decrease, and (2) a parallel release of Na+, K+, P (presumably as negatively charged phosphates), and subsequently Cl- from cytoplasm and the cell as a whole. In addition P, Zn2+, and subsequently Ca2+ increase in acidocalcisomes as Cl- content declines in this compartment. This evidence is the first to document subcellular translocation of, and thus a potential role for, zinc in volume regulatory responses. These coordinated changes in organic and inorganic osmolytes demonstrate that amastigote subcellular compartments, particularly acidocalcisomes, function in maintaining ionic homeostasis in the response of Leishmania amastigotes to hypo-osmotic stress.

We examine representative, group-level wage settlement data to augment the debate on nominal pay rigidity in Britain. We investigate the characteristics of groups that settle at zero and the role of within-firm and external influences. Nominal settlement cuts are rare. Zero nominal wage settlements are more common, but still relatively unusual, highest during (low-inflation) recessionary periods. Small groups, above all firms under duress, appear most likely to settle at zero. Once a group settles at zero it is unlikely to do so again in the short run.

Chronic berylliosis is an uncommon disease that is caused by the inhalation of beryllium particles, dust, or fumes. The distinction between chronic berylliosis and sarcoidosis can be difficult both clinically and histologically, as both entities can have similar presentations and exhibit nonnecrotizing granulomatous inflammation of the lungs. The diagnosis of chronic berylliosis relies on a history of exposure to beryllium, roentgenographic evidence of diffuse nodular disease, and demonstration of beryllium hypersensitivity by ancillary studies, such as lymphocyte proliferation testing. Additional support may be gained by the demonstration of beryllium in lung tissue. Unlike other exogenous particulates, such as asbestos, detection of beryllium in human lung tissue is problematic. The low atomic number of beryllium usually makes it unsuitable for conventional microprobe analysis. We describe a case of chronic berylliosis in which beryllium was detected in lung tissue using atmospheric thin-window energy-dispersive X-ray analysis (ATW EDXA). A woman with a history of occupational exposure to beryllium at a nuclear weapons testing facility presented with progressive cough and dyspnea and a nodular pattern on chest roentgenograph. Open lung biopsy showed nonnecrotizing granulomatous inflammation that was histologically indistinguishable from sarcoidosis. Scanning electron microscopy and ATW EDXA demonstrated particulates containing beryllium within the granulomas. This application of EDXA offers significant advantages over existing methods of beryllium detection in that it is nondestructive, more widely available, and can be performed using routine paraffin sections.

Deregulation of the system of pay determination in Britain was started in 1979 with the removal of incomes policy. The objective was to give employers the freedom to determine wage increases without the restrictions of pay norms or statutory limits. Instead, companies would be able to link changes in pay to the fortunes of the individual enterprise or establishment. By the mid-1990s, had these attempts to decentralize wage negotiations changed the determinants of wage settlement outcomes in Britain? We address the influence of industrial relations institutions and labour market pressures on wage increases between 1979 and 1994 using evidence from the CBI’s Pay Databank. Despite the direction of the Conservative Government’s policy, the external institutional forces of the labour market, particularly the rate of inflation and comparability, appear to exert an enduring influence, both qualitatively and quantitatively, on pay determination.

Barnacles deposit a thin layer of calcium carbonate at the base of the shell in an incremental fashion. The shells are therefore well suited to provide a temporal record of metal exposure, if metals from the water are incorporated into the shell as growth increments form. Fine temporal resolution of metal exposure would prove useful for studying environmental exposure or biomineralization processes. For this study, barnacles were exposed to artificial sea water (ASW) enriched with either strontium, manganese or lead. Barnacles were exposed to different levels of Sr and Mn for different lengths of time from 2 h to 2 d. The elemental composition of the shells was determined by electron probe X-ray microanalysis (EPXMA) imaging. Strontium and Mn were incorporated into the shells in direct proportion to the metal levels in the ASW. A 2 h pulse of 1 mM Mn-enriched ASW did not result in a detectable band of Mn in the shell; however, a 6-h exposure yielded a clearly visible band of Mn-rich shell. Exposure to 0.24 μM Pb-enriched ASW caused marked reduction in the Ca content of the shell; however, a Pb-rich growth band was not detected in the shell. Barnacles analysed by EPXMA imaging provide a sequential record of metal exposure for Mn and Sr, with a time resolution of approximately 6 h. Enrichment of ASW with either Sr or Mn can be used as a probe to monitor biomineralization events.

Analyse de la distinction entre l'art et l'artisanat etablie par R. G. Collingwood dans son ouvrage intitule «The principles of art» (1938). Examinant la definition de l'art comme expression imaginative et comme activite d'externalisation, l'A. montre que l'artisanat participe de l'art en ce sens que l'habilete technique de l'artiste est le moyen de realiser la fin de l'oeuvre dans la communication avec le public

Hexakis (2-methoxyisobutylisonitrile) technetium-99m (99mTc-MIBI) is a gamma-emitting radiopharmaceutical probe currently in clinical use to evaluate myocardial perfusion. Biochemical and cellular pharmacological studies have suggested that Tc-MIBI, a lipophilic cation, is sequestered in mitochondria in response to transmembrane potentials. To assess directly the subcellular distribution of the probe in heart tissue, cultured chick heart cells were analyzed by electron-probe X-ray microanalysis (EPXMA) following equilibration in micromolar concentrations of carrier-added 99Tc-MIBI, the ground-state radiopharmaceutical. Quantitation of the physiological elements Na, Ca, Mg, K, S, P, and Cl was correlated with exposure to increasing concentrations of 99Tc-MIBI. EPXMA signals indicated that 99Tc-MIBI was concentrated up to 1,000 times into mitochondria in a dose-dependent fashion based on measured Tc content in the mitochondria. Inner membrane potential (delta psi) of individual mitochondria was calculated as -117 mV using the Nernst equation. Concentrations of 99Tc-MIBI > 36 microM caused a significant efflux of K and Mg from the cell, as well as an increase in Cl in the mitochondria. Comparison of cell ultrastructure with conventional electron microscopy at extracellular 99Tc-MIBI concentrations of 36-72 microM showed no changes compared with control. 99Tc-MIBI allows valuable in situ investigation of cellular bioenergetics with EPXMA by quantitation of delta psi.

The reliability of power electronic devices is significantly related to the material properties of the applied substrates which carry the semiconductor chip and the electric interconnections. The most common solution to fulfill the stringent requirements of these devices, with respect to high isolation voltage, good thermal conductivity, high temperature cycling reliability and low cost, is to use ceramic substrates with copper layers on both sides. However, the currently increasing reliability standards in power electronics lead to a situation where common DCB substrates reach their limits in meeting these higher requirements.

Background information. Poly P (inorganic polyphosphate) is a polymer formed by Pi residues linked by high-energy phosphoanhydride bonds. The presence of poly P in bacteria, fungi, algae and protists has been widely recognized, but the distribution of poly P in more complex eukaryotes has been poorly studied. Poly P accumulates, together with calcium, in acidic vesicles or acidocalcisomes in a number of organisms and possesses a diverse array of functions, including roles in stress response, blood clotting, inflammation, calcification, cell proliferation and apoptosis. Results. We report here that a considerable amount of phosphorus in the yolk of chicken eggs is in the form of poly P. DAPI (4′,6-diamidino-2-phenylindole) staining showed that poly P is localized mainly in electron-dense vesicles located inside larger vacuoles (compound organelles) that are randomly distributed in the yolk. These internal vesicles were shown to contain calcium, potassium, sodium, magnesium, phosphorus, chlorine, iron and zinc, as detected by X-ray microanalysis and elemental mapping. These vesicles stain with the acidophilic dye Acridine Orange. The presence of poly P in organellar fractions of the egg yolk was evident in agarose gels stained with Toluidine Blue and DAPI. Of the total phosphate (Pi) of yolk organelles, 16% is present in the form of poly P. Total poly P content was not altered during the first 4 days of embryogenesis, but poly P chain length decreased after 1 day of development. Conclusions. The results of the present study identify a novel organelle in chicken egg yolk comprising acidic vesicles with a morphology, physiology and composition similar to those of acidocalcisomes, within larger acidic vacuoles. The elemental composition of these acidocalcisomes is proportionally similar to the elemental composition of the yolk, suggesting that most of these elements are located in these organelles, which might be an important storage compartment in eggs.

Two cases of pulmonary alveolar proteinosis, including one death, occurred in workers at a facility producing indium-tin oxide (ITO), a compound used in recent years to make flat panel displays. Both workers were exposed to airborne ITO dust and had indium in lung tissue specimens. One worker was tested for autoantibodies to granulocytemacrophage-colonystimulating factor (GM-CSF) and found to have an elevated level. These cases suggest that inhalational exposure to ITO causes pulmonary alveolar proteinosis, which may occur via an autoimmune mechanism.

The 1980s witnessed a considerable increase in the rate of growth of manufacturing productivity in Britain. This paper attempts to reveal the extent of systematic change at the work-place which was associated with this improvement. The study focuses on changes in working practices introduced concurrently with negotiations over wage increases; in every year throughout the decade around one third of wage settlements involving trade unions included productivity enhancing change. It is argued that the discipline of competitive pressure and the absence of incomes policy gave rise to the growth in the incidence of wage negotiations which featured a productivity element.

A novel preparation of spontaneously contracting heart cells embedded in a collagen strand provides an ideal experimental system for correlative structure-function experiments that utilize the techniques of electron microscopy, quantitative electron probe x-ray microanalysis (EPXMA) and imaging. Heart cells grown within the strand for 1 day possess the subcellular content and distribution of physiologically relevant elements—Na, Mg, P, 5, Cl, K, and Ca—found in intact heart cell preparations. The presence of junctional specializations between, and organized myofibrils within, the majority of cells after 1 day in culture also establishes that the collagen matrix promotes vigorous cell development as well as maintains physiological integrity. EPXMA, combined with ultrastructural analyses, provides elemental content data on a cell-by-cell basis. In studies presented here, viable cells, comprising over 80% of the strand cell population, could be distinguished easily from those which had been functionally compromised, not only by aberrant structure but also by altered subcellular compartmentation of Na, K, Cl, and Ca. Within individual viable cells, compartmental differences in element content were notable especially between mitochondria and cytoplasm. However, nuclear euchromatin, but not heterochromatin, appeared approximately identical to cytoplasm in elemental and water content. In such cells, the cytoplasmic K:Na ratio was maintained at a high level (∼15:1). The results with respect to K, Na, and other elements demonstrated the integrity of membrane transport mechanisms regulating the movement and distribution of ions and the maintenance of ionic homeostasis in cells of the strand preparation.

The purpose of this paper is to examine the pattern wamed in 1985. and outcomes of wage settlements in British There seems to be neither a social nor economic rationale manufacturing industry during the 1980s. The period for the present level of pay settlements. The combination since 1979 represents the longest sustained period °f very unemployment and a union movement in of free collective bargaining in the post-war years, disarray might have been expected to cause manage With the removal of explicit norms or guide-posts ment t0 lake a ralher hard nosed attitude t0 labour to the outcome of periodic negotiations over wage costs" (Financial Times Leader' 'The Pantomime over Pnv' ? Nnvpmhpr 1 changes, the pattern of wage increases generated by y ' the process of company and plant level pay settlements This paper addresses the outcomes of pay determi should reveal a great deal about the behaviour of nation since the return to free collective bargaining and influences that bear on pay determination, and seeks to focus on the influences impacting on, Despite the absence of incomes policy throughout or the rationale for, those outcomes and looks at the 1980s the outcome of wage negotiations none how these have changed over the last decade.

3-Methoxybenzamide is a weak inhibitor of the essential bacterial cell division protein FtsZ. Exploration of the structure-activity relationships of 3-methoxybenzamide analogues led to the identification of potent anti-staphylococcal compounds.

The general trend of microscopical investigation in biology from the 1950’s to the early 1970’s was towards obtaining structural information. This goal initially was met using heavy metal and/or aldehyde fixatives, room temperature dehydration with polar organic liquids, embedding with epoxy and acrylate resins, and thin sectioning at room temperature. By the mid 1970’s, a perceptible change occurred in the direction of both light and electron microscopy towards investigation of the chemical reactivity and composition of structures made visible with increasingly better spatial resolution for light and electron microscopes. Between the mid 1980’s and the present, innovations in microanalytical techniques, including x-ray microscopy and microanalysis, secondary ion mass spectrometry, laser microprobe mass analysis, the scanning probe microscopies, and confocal/multi-photon microscopy, and development of compounds for visualization of molecular and ionic sites within individual living cells as well as membranes, redefined the goal of microscopical preservation: to stabilize cell structure and composition as they exist in the living state.

The current trend of x-ray mapping in Electron Probe X-ray Microanalysis (EPXMA) is toward full spectrum collection, or spectrum imaging [1]. Spectrum imaging is the collection and spatial registration of all x-ray events, yielding a spectral data cube. Recent developments in spectral analytical techniques have greatly improved the efficiency and precision of data handling from quantitative chemical images; principal component and cluster analysis has significantly aided in unambiguous identification and assessment of localized data as well as the reduction of statistical noise in the images; fitting of all elemental peaks at each pixel to standards accomplishes a similar result as well as providing real-time or off-line quantitative images; “programmed beam acquisition” (PBA) enables the exciting beam to only dwell for sufficient times per pixel in regions of the specimen necessary to acquire a predetermined statistical precision (e.g.±2 standard deviations). PBA, while not a new concept [1,2], is rarely used. However it is of particular importance to biological samples, which are sensitive to beam damage and inherently produce low x-ray count rates.

Leishmania major promastigotes contain electron-dense vacuoles. The elemental composition of these vacuoles and of the cytoplasm was measured by electron probe X-ray microanalysis, using rapid cryopreservation techniques to prevent alterations in composition due to diffusion. The electron-dense vacuoles are rich in P, presumably present as polyphosphate (poly P). Mg is present at about 9 times its cytoplasmic level. There is sufficient Mg to largely neutralize most of the negative charge of the Poly P. The electron-dense vacuoles also contain appreciable amounts of Ca and Zn, which are not detectable in the cytoplasm, as well as Na, K, and Cl, the latter two at concentrations below that of the cytoplasm. These results suggest that the vacuolar membranes have at least one cation transport system. Incubation of the promastigotes for 1 h in the absence of phosphate in the presence or absence of glucose did not cause significant changes in the vacuolar contents of P, Mg, or Zn, but changes in K and Cl content were observed in both the electron-dense vacuoles and in the cytoplasm.

This paper presents a broad survey of the rationale for electron probe X-ray microanalysis (EPXMA) and the various methods for obtaining qualitative and quantitative information on the distribution and amount of elements, particularly calcium, in cryopreserved cells and tissues. Essential in an introductory consideration of microanalysis in biological cryosections is the physical basis for the instrumentation, fundamentals of X-ray spectrometry, and various analytical modes such as static probing and X-ray imaging. Some common artifacts are beam damage and contamination. Inherent pitfalls of energy dispersive X-ray systems include Si escape peaks, doublets, background, and detector calibration shifts. Quantitative calcium analysis of thin cryosections is carried out in real time using a multiple least squares fitting program on filtered X-ray spectra and normalizing the calcium peak to a portion of the continuum. Recent work includes the development of an X-ray imaging system where quantitative data can be retrieved off-line. The minimum detectable concentration of calcium in biological cryosections is approximately 300 mumole kg dry weight with a spatial resolution of approximately 100 A. The application of electron energy loss (EELS) techniques to the detection of calcium offers the potential for greater sensitivity and spatial resolution in measurement and imaging. Determination of mass thickness with EELS can facilitate accurate calculation of wet weight concentrations from frozen hydrated and freeze-dried specimens. Calcium has multiple effects on cell metabolism, membrane transport and permeability and, thus, on overall cell physiology or pathophysiology. Cells can be rapidly frozen for EPXMA during basal or altered functional conditions to delineate the location and amount of calcium within cells and the changes in location and concentration of cations or anions accompanying calcium redistribution. Recent experiments in our laboratory document that EPXMA in combination with other biochemical and electrophysiological techniques can be used to study, for example, sodium and calcium compartmentation in cultured cardiac cells. Such analyses can also be used to clarify the role of calcium in anoxic renal cell injury and to evaluate proposed ionic defects in cells of individuals with cystic fibrosis.

The protozoan parasite Leishmania donovani encounters large fluctuations in osmolality as it cycles between its insect vector and human host. The flagellated promastigote exhibits regulatory volume responses involving organic and inorganic osmolytes, but little is known about volume regulation in the clinically relevant amastigote that multiplies within the parasitophorous vacuoles of mammalian host cells. Using a combination of morphological, X-ray microanalytical, and biochemical approaches we determined that non-motile amastigotes respond to hypotonic stress with (1) an amino acid and l-alanine-mediated regulatory volume decrease, and (2) a parallel release of Na+, K+, P (presumably as negatively charged phosphates), and subsequently Cl- from cytoplasm and the cell as a whole. In addition P, Zn2+, and subsequently Ca2+ increase in acidocalcisomes as Cl- content declines in this compartment. This evidence is the first to document subcellular translocation of, and thus a potential role for, zinc in volume regulatory responses. These coordinated changes in organic and inorganic osmolytes demonstrate that amastigote subcellular compartments, particularly acidocalcisomes, function in maintaining ionic homeostasis in the response of Leishmania amastigotes to hypo-osmotic stress.

Against a background of increased decentralisation in the structure of wage decision making, we analyse the effects of unions on the dispersion and persistence of pay settlements over the medium term using a longitudinal data set covering British private sector establishments over the period 1987-2001. It seems that the union effect of a reduction in wage dispersion in pay levels observed in earlier studies is repeated when we follow wage changes (settlements) over the medium term. Declining union presence seems therefore to account for some of the increase in longer-term wage dispersion over the sample period. The increase in aggregate wage settlement dispersion seems to have been accompanied by an increase in the permanent rather than transitory components of the variance and this stems mostly from the non-union sector.

The number of strikes reported in British industry has been on a downward trend over the past two decades, falling in 1998 to its lowest level since records began. This may indicate that relations within British industry have improved; however, the same period has also witnessed a sharp increase in the number of individual ACAS and employment tribunal cases. We discuss possible reasons for the changes in the patterns of industrial unrest over time and use individual micro-data to examine whether the observed decline in strike activity has actually been associated with an improvement in perceptions of workplace industrial relations.

The elemental composition of the cytoplasm, electron dense vacuoles, and heterochromatin and euchromatin regions of the nucleus of Leishmania major promastigotes was measured by electron probe X-ray microanalysis under iso-osmotic conditions (305 mOsM) and shortly after a sudden increase (to 615 mOsM) or decrease (to 153 mOsM) in the osmolality of the buffer in which they were suspended. In response to acute hypotonicity a complete loss of Na from the electron dense vacuoles and an approximately threefold decrease in the Na concentrations in the cytoplasm and the nuclear regions occurred, together with an approximately threefold decrease in Cl content in each compartment and a smaller (approx. 1.2-fold) decrease in K content. Thus, in addition to the rapid change in shape and release of amino acids known to occur in response to acute hypo-osmotic stress, a major efflux of Na and Cl, and, to a lesser extent, of K, also occurs. In response to acute hypertonicity Na in the acidocalcisomes did not change but Na content of the cytoplasm decreased by 33%. A small increase in the S content of the cytoplasm and the electron dense vacuolar compartments occurred. No changes were detectable in Ca or Zn content in any of the compartments examined in response to hypotonicity or hypertonicity.

Electron microscopy is analytical to some extent. Analytical electron microscopy (AEM) can be defined as the observation of the interaction of electrons with matter at a resolution smaller than that which could be seen with the naked eye. This obviously includes all images in general; however, the term “microprobe analysis” has evolved to make it essentially a form of microchemistry in particular. The wavelength of X-rays can be measured using a wavelength dispersive spectrometer, or the characteristic energies can be measured by means of an energy dispersive spectrometer. Each instrumental configuration has its own advantages and limitations, and the choice of instrumentation depends not only on user preference but also on the particular task at hand. For example, analytical transmission electron microscopy can allow the user to obtain crystallographic information about the sample by means of selected area electron diffraction, whereas analytical scanning electron microscopy permits the elemental analysis of bulk samples and, more recently, the potential to determine the microcrystallographic characteristics of mineral specimens. This approach is particularly useful for the analysis of clinical specimens when combined with backscattered electron imaging, which enhances the ability to detect inorganic particulates of relatively high atomic number in an organic matrix of low atomic number.

This chapter explains how biological material can be preserved in the quasi in vivo state at very high spatial and temporal resolution for structure. The basis for the unique efficacy of electron probe X-ray microanalysis (EPXMA) in identifying the distribution of elements in tissues and cells, presumably close to the in vivo state, is the method known as cryopreservation. Cryopreservation, beginning with quick freezing, is imperative for most biological applications of EPXMA to permanently immobilize elements in a given specimen while preserving cell structure close to the in vivo state. To that end, it must be capable of eliminating or at least minimizing untoward ice crystal growth that causes cryoartifacts. The objective of quick freezing is to withdraw heat from the specimen at a very fast rate. The major factors to be considered toward that goal are the nature of the coolant and the geometry of the specimen with its holder. The coolant is either solid (for example, copper for impact) or liquid (for example, propane for immersion) and must have large heat capacity and high conductivity at very low temperatures.

This chapter outlines the major preparative techniques employed to date for qualitative energy dispersive X-ray analysis in human pathology. In problems of human pathology, much of the microanalytical work to date has relied on relatively unsophisticated techniques of specimen preparation. These methods require only minor modifications from routine histologic processing for light or electron microscopy. Because most clinical cases in which analytical electron microscopy (AEM) has been used have involved foreign material identification and localization, usually nondiffusible, inorganic xenobiotics, these simple techniques have usually been adequate. Simple preparative techniques are generally superior to more elaborate ones if they are sufficient to resolve the problem under study. Undoubtedly, greater numbers of pathologists in the future will address more difficult questions in human pathophysiology using microanalytical techniques to describe the in vivo distribution of soluble endogenous elements and molecules. Such work necessarily calls for significant technical improvements in and wider adoption of more elaborate methods of tissue preparation, such as cryotechniques.

Energy dispersive X-ray (EDX) for characterizing foreign matter within human tissue is most frequently employed in the study of inhaled particles within the substance of the lung. The most common diagnostic role of analytic electron microscopy is seen in studies of the pneumoconioses: the spectrum of nonneoplastic reactions that the lung parenchyma displays toward inhaled and retained minerals, metals, dusts, and organic materials. This varied family of diseases includes asbestosis, silicosis, coal workers' pneumoconiosis, hard metal pneumoconiosis, and many others. A variety of metals and other elements may be detected with EDX analysis, including copper in tissues of patients with Wilson's disease, thorium in patients who have been injected in the past with the radiographic contrast material Thorotrast, gold in patients treated with long-term chrysotherapy, and arsenic or lead in patients with heavy metal poisoning. Analytic electron microscopy may corroborate or refute a known or suspected exposure, which may be remote. The rare earths cerium and lanthanum have been detected in alveolar macrophages and in lung tissue at least 15 years after the last known exposure, suggesting a means for their metabolism and biopersistence.

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Cell physiological and pathophysiological studies often require information about the elemental composition of intracellular organelles in situ. Electron probe X-ray microanalysis (EPXMA) is one of the few methods by which intracellular elemental content and distribution can be measured simultaneously. While several cryofixation techniques for EPXMA have been utilized on isolated cells, few have been applied successfully to whole tissue in vivo or in situ. A recently developed, commercial, portable, metal-mirror device was used for preserving kidney in situ to determine the intracellular element distribution in proximal tubule cells. Kidneys of male rats were exposed, cryofixed, and analyzed for organelle elemental contents by EPXMA imaging. In addition, some portions of the frozen tissue were prepared for conventional transmission electron microscopy. Proximal tubules were preserved with intact brush borders and open lumens. The quality of preservation of tubule cell organelles varied inversely as a function of depth from the point of first contact with the mirror surface; the best preservation was within 15 microns, while the poorest preservation was deeper than 30 microns. Analysis of EPXMA images from the best-preserved regions revealed that proximal tubule cell cytoplasmic K/Na was approximately 6, cytoplasmic Cl was low relative to other subcellular compartments, and mitochondrial Ca levels were 1.8 nmole/mg dry weight; these observations indicate that the cells were physiologically viable at the time of cryofixation. The advantages of in situ cryofixation by this metal-mirror method include acquisition of organelle elemental content data in vivo, ease of use, reproducibility, portability, applicability to other tissues, and suitability for pathophysiological studies.