Spontaneous diabetes mellitus in transgenic mice expressing human islet amyloid polypeptide.

The islet in non-insulin-dependent diabetes mellitus (NIDDM) is characterized by loss of beta cells and large local deposits of amyloid derived from the 37-amino acid protein, islet amyloid polypeptide (IAPP). We have hypothesized that IAPP amyloid forms intracellularly causing beta-cell destruction under conditions of high rates of expression. To test this we developed a homozygous transgenic mouse model with high rates of expression of human IAPP. Male transgenic mice spontaneously developed diabetes mellitus by 8 weeks of age, which was associated with selective beta-cell death and impaired insulin secretion. Small intra- and extracellular amorphous IAPP aggregates were present in islets of transgenic mice during the development of diabetes mellitus. However, IAPP derived amyloid deposits were found in only a minority of islets at approximately 20 weeks of age, notably after development of diabetes mellitus in male transgenic mice. Approximately 20% of female transgenic mice spontaneously developed diabetes mellitus at 30+ weeks of age, when beta-cell degeneration and both amorphous and amyloid deposits of IAPP were present. We conclude that overexpression of human IAPP causes beta-cell death, impaired insulin secretion, and diabetes mellitus. Large deposits of IAPP derived amyloid do not appear to be important in this cytotoxicity, but early, small amorphous intra- and extracellular aggregates of human IAPP were consistently present at the time of beta-cell death and therefore may be the most cytotoxic form of IAPP.

Psychophysical evidence for fast region-based segmentation processes in motion and color.

Theories of image segmentation suggest that the human visual system may use two distinct processes to segregate figure from background: a local process that uses local feature contrasts to mark borders of coherent regions and a global process that groups similar features over a larger spatial scale. We performed psychophysical experiments to determine whether and to what extent the global similarity process contributes to image segmentation by motion and color. Our results show that for color, as well as for motion, segmentation occurs first by an integrative process on a coarse spatial scale, demonstrating that for both modalities the global process is faster than one based on local feature contrasts. Segmentation by motion builds up over time, whereas segmentation by color does not, indicating a fundamental difference between the modalities. Our data suggest that segmentation by motion proceeds first via a cooperative linking over space of local motion signals, generating almost immediate perceptual coherence even of physically incoherent signals. This global segmentation process occurs faster than the detection of absolute motion, providing further evidence for the existence of two motion processes with distinct dynamic properties.

A biomarker that identifies senescent human cells in culture and in aging skin in vivo.

Normal somatic cells invariably enter a state of irreversibly arrested growth and altered function after a finite number of divisions. This process, termed replicative senescence, is thought to be a tumor-suppressive mechanism and an underlying cause of aging. There is ample evidence that escape from senescence, or immortality, is important for malignant transformation. By contrast, the role of replicative senescence in organismic aging is controversial. Studies on cells cultured from donors of different ages, genetic backgrounds, or species suggest that senescence occurs in vivo and that organismic lifespan and cell replicative lifespan are under common genetic control. However, senescent cells cannot be distinguished from quiescent or terminally differentiated cells in tissues. Thus, evidence that senescent cells exist and accumulate with age in vivo is lacking. We show that several human cells express a beta-galactosidase, histochemically detectable at pH 6, upon senescence in culture. This marker was expressed by senescent, but not presenescent, fibroblasts and keratinocytes but was absent from quiescent fibroblasts and terminally differentiated keratinocytes. It was also absent from immortal cells but was induced by genetic manipulations that reversed immortality. In skin samples from human donors of different age, there was an age-dependent increase in this marker in dermal fibroblasts and epidermal keratinocytes. This marker provides in situ evidence that senescent cells may exist and accumulate with age in vivo.

Astrocytes and Müller Cell Alterations During Retinal Degeneration in a Transgenic Rat Model of Retinitis Pigmentosa

Purpose: Retinitis pigmentosa includes a group of progressive retinal degenerative diseases that affect the structure and function of photoreceptors. Secondarily to the loss of photoreceptors, there is a reduction in retinal vascularization, which seems to influence the cellular degenerative process. Retinal macroglial cells, astrocytes, and Müller cells provide support for retinal neurons and are fundamental for maintaining normal retinal function. The aim of this study was to investigate the evolution of macroglial changes during retinal degeneration in P23H rats. Methods: Homozygous P23H line-3 rats aged from P18 to 18 months were used to study the evolution of the disease, and SD rats were used as controls. Immunolabeling with antibodies against GFAP, vimentin, and transducin were used to visualize macroglial cells and cone photoreceptors. Results: In P23H rats, increased GFAP labeling in Müller cells was observed as an early indicator of retinal gliosis. At 4 and 12 months of age, the apical processes of Müller cells in P23H rats clustered in firework-like structures, which were associated with ring-like shaped areas of cone degeneration in the outer nuclear layer. These structures were not observed at 16 months of age. The number of astrocytes was higher in P23H rats than in the SD matched controls at 4 and 12 months of age, supporting the idea of astrocyte proliferation. As the disease progressed, astrocytes exhibited a deteriorated morphology and marked hypertrophy. The increase in the complexity of the astrocytic processes correlated with greater connexin 43 expression and higher density of connexin 43 immunoreactive puncta within the ganglion cell layer (GCL) of P23H vs. SD rat retinas. Conclusions: In the P23H rat model of retinitis pigmentosa, the loss of photoreceptors triggers major changes in the number and morphology of glial cells affecting the inner retina.

EU immigration trends and policies: Spain and France as case studies. Jean Monnet/Robert Schuman Paper Series Vol. 13 No. 10, August 2013

From the Introduction. The Treaty on European Union, also known as the Treaty of Maastricht or the Maastricht Treaty, created the European Union (EU) from the existing European Economic Community (EEC.) It was signed by the member states on February 7, 1992, and entered into force on November 1, 1993.1 Among its many innovations was the creation of European citizenship, which would be granted to any person who was a citizen of an EU member state. Citizenship, however, is intertwined with immigration, which the Treaty also attempted to address. Policy on visas, immigration and asylum was originally placed under Pillar 3 of the EU, which dealt with Justice and Home Affairs. In 1997, however, the Amsterdam Treaty moved these policies from Pillar 3 to Pillar 1, signaling “a shift toward more supranational decision-making in this area,” as opposed to the intergovernmental method of Pillars 2 and 3.2

Temporal Control of Leaf Complexity by miRNA-Regulated Licensing of Protein Complexes

The tremendous diversity of leaf shapes has caught the attention of naturalists for centuries. In addition to interspecific and intraspecific differences, leaf morphologies may differ in single plants according to age, a phenomenon known as heteroblasty. In Arabidopsis thaliana, the progression from the juvenile to the adult phase is characterized by increased leaf serration. A similar trend is seen in species with more complex leaves, such as the A. thaliana relative Cardamine hirsuta, in which the number of leaflets per leaf increases with age. Although the genetic changes that led to the overall simpler leaf architecture in A. thaliana are increasingly well understood, less is known about the events underlying age-dependent changes within single plants, in either A. thaliana or C. hirsuta. Here, we describe a conserved miRNA transcription factor regulon responsible for an age-dependent increase in leaf complexity. In early leaves, miR319-targeted TCP transcription factors interfere with the function of miR164-dependent and miR164-independent CUC proteins, preventing the formation of serrations in A. thaliana and of leaflets in C. hirsuta. As plants age, accumulation of miR156-regulated SPLs acts as a timing cue that destabilizes TCP-CUC interactions. The destabilization licenses activation of CUC protein complexes and thereby the gradual increase of leaf complexity in the newly formed organs. These findings point to posttranslational interaction between unrelated miRNA-targeted transcription factors as a core feature of these regulatory circuits.

Europeanization and the inclusive strategies of executive actors

In Europeanized policy domains, executive actors are considered especially powerful because they are directly responsible for international negotiations. However, in order to avoid failing in the ratification process, they are also highly dependent on the support of domestic, non-state actors. We argue that in Europeanized decision-making processes, state actors are not passively lobbied, but actively seek collaboration with - and support from - domestic actors. We apply stochastic actor-based modelling for network dynamics to collaboration data on two successive bilateral agreements on the free movement of persons between Switzerland and the European Union (EU). Results confirm our hypotheses that state actors are not passively lobbied, but actively look for collaboration with other actors, and especially with potential veto players and euro-sceptical actors from both the conservative Right and the Left.

Europeanization, Institutional Changes and Differential Empowerment

The issue of European integration is of utmost importance for contemporary Swiss politics, as underscored by the presence of three decision-making processes relating to bilateral agreements with the EU, and two additional processes with a strong European dimension (the telecommunication act and the immigration law), among the 11 most important processes of the early 2000s. Previous chapters have highlighted substantial differences between domestic and Europeanized decision-making processes in terms of institutional design and decision-making structures. Chapters 2 and 3 suggest that the peculiarities of the three decision-making processes relating to bilateral agreements go along with specific power configurations among political actors. Chapter 5 draws our attention to the impact of Europeanization on the specific decision-making structure at work in a given policy process.

Use of space by domestic chicks housed in complex aviaries.

To improve the understanding of the development of locomotor capacity in layer hens, we measured how female laying hen chicks (n=120) of four different strains (LSL-lite, Hyline Brown, Dekalb White, Lohmann Brown; 3 groups of 10 chicks per line) utilized the ground, the air, elevated horizontal (platforms and perches) and inclined surfaces (ramps and ladders) in an aviary until 9 weeks of age. We used infra-red video recordings to perform all-occurrences sampling of locomotive behavioural and perching events that occurred on the ground—where bedding material, food and water were provided, in the air, and on elevated horizontal and inclined surfaces within weekly 30-min sampling periods. Chicks preferred level ground during the first week of life compared to weeks 5–9 (P<0.0001) and performed 52% of all behavioural events in this section. Elevated surface use began at 2 weeks of age and increased over time (P=0.003), where most behaviour was performed in S2 (45% of all events). Chicks preferred horizontal to inclined surfaces, which were used from weeks 2–5 with maximum use occurring during weeks 2 and 3. Lohmann LSL chicks used the space above the ground most frequently (P=0.05) and performed more aerial ascent/descent behaviour than other lines (P<0.0001). Overall activity levels declined with age (P<0.0001). In summary layer chicks almost exclusively locomoted on the ground but utilized elevated horizontal surfaces (perch, first platform) as early as 2 weeks. These results provide information for improving space use in rearing aviaries by introducing lower perches, platforms and ramps/ladders to accommodate age-dependent locomotor abilities.

Mitochondrial impairments contribute to Spinocerebellar ataxia type 1 progression and can be ameliorated by the mitochondria-targeted antioxidant MitoQ

Spinocerebellar ataxia type 1 (SCA1), due to an unstable polyglutamine expansion within the ubiquitously expressed Ataxin-1 protein, leads to the premature degeneration of Purkinje cells (PCs), decreasing motor coordination and causing death within 10-15 years of diagnosis. Currently, there are no therapies available to slow down disease progression. As secondary cellular impairments contributing to SCA1 progression are poorly understood, here, we focused on identifying those processes by performing a PC specific proteome profiling of Sca1154Q/2Q mice at a symptomatic stage. Mass spectrometry analysis revealed prominent alterations in mitochondrial proteins. Immunohistochemical and serial block-face scanning electron microscopy analyses confirmed that PCs underwent age-dependent alterations in mitochondrial morphology. Moreover, colorimetric assays demonstrated impairment of the electron transport chain complexes (ETC) and decrease in ATPase activity. Subsequently, we examined whether the mitochondria-targeted antioxidant MitoQ could restore mitochondrial dysfunction and prevent SCA1-associated pathology in Sca1154Q/2Q mice. MitoQ treatment both presymptomatically and when symptoms were evident ameliorated mitochondrial morphology and restored the activities of the ETC complexes. Notably, MitoQ slowed down the appearance of SCA1-linked neuropathology such as lack of motor coordination as well as preventing oxidative stress-induced DNA / RNA damage and PC loss. Our work identifies a central role for mitochondria in PC degeneration in SCA1 and provides evidence for the supportive use of mitochondria-targeted therapeutics in slowing down disease progression.

Centennial-scale evolution of Dansgaard-Oeschger events in sediment core MD95-2006

There is much uncertainty surrounding the mechanisms that forced the abrupt climate fluctuations found in many palaeoclimate records during Marine Isotope Stage (MIS)-3. One of the processes thought to be involved in these events is the Atlantic Meridional Overturning Circulation (MOC), which exhibited large changes in its dominant mode throughout the last glacial period. Giant piston core MD95-2006 from the northeast Atlantic Ocean records a suite of palaeoceanographic proxies related to the activity of both surface and deep water masses through a period of MIS-3 when abrupt climate fluctuations were extremely pronounced. A two-stage progression of surface water warming during interstadial warm events is proposed, with initial warming related to the northward advection of a thin warm surface layer within the North Atlantic Current, which only extended into deeper surface layers as the interstadial progressed. Benthic foraminifera isotope data also show millennial-scale oscillations but of a different structure to the abrupt surface water changes. These changes are argued to partly be related to the influence of low-salinity deepwater brines. The influence of deepwater brines over the site of MD95-2006 reached a maximum at times of rapid warming of surface waters. This observation supports the suggestion that brine formation may have helped to destabilize the accumulation of warm, saline surface waters at low latitudes, helping to force the MOC into a warm mode of operation. The contribution of deepwater brines relative to other mechanisms proposed to alter the state of the MOC needs to be examined further in future studies.

Ring Out Your Dead : Distribution, form, and function of iron amulets in the late Iron Age grave fields of Lovö

The purpose of this study is to analyze the distribution, forms, and function(s) of iron amulets deposited in the late Iron Age gravefields of Lovö, with the goal of ascertaining how (and so far as possible why) these objects were utilized in rituals carried out during and after burials. Particular emphasis is given to re-interpreting the largest group of iron amulets, the iron amulet rings, in a more relational and practice-focused way than has heretofore been attempted. By framing burial analyses, questions of typology, and evidence of ritualized actions in comparison with what is known of other cult sites in Mälardalen specifically– and theorized about the cognitive landscape(s) of late Iron Age Scandinavia generally– a picture of iron amulets as inscribed objects made to act as catalytic, protective, and mediating agents is brought to light.

Population pharmacokinetic analysis of mycophenolic acid in renal transplant recipients following oral administration of mycophenolate mofetil

Aim To develop a population pharmacokinetic model for mycophenolic acid in adult kidney transplant recipients, quantifying average population pharmacokinetic parameter values, and between- and within-subject variability and to evaluate the influence of covariates on the pharmacokinetic variability. Methods Pharmacokinetic data for mycophenolic acid and covariate information were previously available from 22 patients who underwent kidney transplantation at the Princess Alexandra Hospital. All patients received mycophenolate mofetil 1 g orally twice daily. A total of 557 concentration-time points were available. Data were analysed using the first-order method in NONMEM (version 5 level 1.1) using the G77 FORTRAN compiler. Results The best base model was a two-compartment model with a lag time (apparent oral clearance was 271 h(-1), and apparent volume of the central compartment 981). There was visual evidence of complex absorption and time-dependent clearance processes, but they could not be successfully modelled in this study. Weight was investigated as a covariate, but no significant relationship was determined. Conclusions The complexity in determining the pharmacokinetics of mycophenolic acid is currently underestimated. More complex pharmacokinetic models, though not supported by the limited data collected for this study, may prove useful in the future. The large between-subject and between-occasion variability and the possibility of nonlinear processes associated with the pharmacokinetics of mycophenolic acid raise questions about the value of the use of therapeutic monitoring and limited sampling strategies.

Effect of aldosterone antagonism on myocardial dysfunction in hypertensive patients with diastolic heart failure

Background - Specific treatments targeting the pathophysiology of hypertensive heart disease are lacking. As aldosterone has been implicated in the genesis of myocardial fibrosis, hypertrophy, and dysfunction, we sought to determine the effects of aldosterone antagonism on myocardial function in hypertensive patients with suspected diastolic heart failure by using sensitive quantitative echocardiographic techniques in a randomized, double-blinded, placebo-controlled study. Methods and Results - Thirty medically treated ambulatory hypertensive patients (19 women, age 62 +/- 6 years) with exertional dyspnea, ejection fraction >50%, and diastolic dysfunction (E/A 250m/sec) and without ischemia were randomized to spironolactone 25 mg/d or placebo for 6 months. Patients were overweight (31 +/- 5 kg/m(2)) with reduced treadmill exercise capacity (6.7 +/- 2.1 METS). Long-axis strain rate (SR), peak systolic strain, and cyclic variation of integrated backscatter (CVIB) were averaged from 6 walls in 3 standard apical views. Mean 24-hour ambulatory blood pressure at baseline (133 +/- 17/80 +/- 7mm Hg) did not change in either group. Values for SR, peak systolic strain, and CVIB were similar between groups at baseline and remained unchanged with placebo. Spironolactone therapy was associated with increases in SR (baseline: -1.57 +/- 0.46 s(-1) versus 6-months: -1.91 +/- 0.36 s(-1), P < 0.01), peak systolic strain (-20.3 &PLUSMN; 5.0% versus -26.9 &PLUSMN; 4.3%, P < 0.001), and CVIB (7.4 +/- 1.7dB versus 8.6 +/- 1.7 dB, P = 0.08). Each parameter was significantly greater in the spironolactone group compared with placebo at 6 months (P = 0.05, P = 0.02, and P = 0.02, respectively), and the increases remained significant after adjusting for baseline differences. The increase in strain was independent of changes in blood pressure with intervention. The spironolactone group also exhibited reduction in posterior wall thickness (P = 0.04) and a trend to reduced left atrial area (P = 0.09). Conclusions - Aldosterone antagonism improves myocardial function in hypertensive heart disease.

Patients with a hypertensive response to exercise have impaired systolic function without diastolic dysfunction or left ventricular hypertrophy

OBJECTIVES We sought to determine if a hypertensive response to exercise (HRE) is associated with myocardial changes consistent with early hypertensive heart disease. BACKGROUND An HRE predicts the development of chronic hypertension (HT) and may reflect a preclinical stage of HT. METHODS Patients with a normal left ventricular (LV) ejection fraction and a negative stress test were recruited into three matched groups: 41 patients (age 56 +/- 10 years) with HRE (210/105 mm Hg in men; > 190/105 in women), comprising 22 patients with (HT+) and 19 without resting hypertension (HT-); and 17 matched control subjects without HRE. Long-axis function was determined by measurement of the strain rate (SR), peak systolic strain, and cyclic variation (CV) of integrated backscatter in three apical views. RESULTS An HRE was not associated with significant differences in LV mass index. Exercise performance and diastolic function were reduced in HRE(HT+) patients, but similar in HRE(HT-) patients and controls. Systolic dysfunction (peak systolic strain, SR, and CV) was significantly reduced in HRE patients (p < 0.001 for all). These reductions were equally apparent in patients with and without a history of resting HT (p = NS) and were independent of LV mass index and blood pressure (p < 0.01). CONCLUSIONS An HRE is associated with subtle systolic dysfunction, even in the absence of resting HT. These changes occur before the development of LV hypertrophy or detectable diastolic dysfunction and likely represent early hypertensive heart disease. (C) 2004 by the American College of Cardiology Foundation.