Morphological and Functional Association of Sec22b/ERS-24 with the pre-Golgi Intermediate Compartment

Yeast Sec22p participates in both anterograde and retrograde vesicular transport between the endoplasmic reticulum (ER) and the Golgi apparatus by functioning as a v-SNARE (soluble N-ethylmaleimide-sensitive factor [NSF] attachment protein receptor) of transport vesicles. Three mammalian proteins homologous to Sec22p have been identified and are referred to as Sec22a, Sec22b/ERS-24, and Sec22c, respectively. The existence of three homologous proteins in mammalian cells calls for detailed cell biological and functional examinations of each individual protein. The epitope-tagged forms of all three proteins have been shown to be primarily associated with the ER, although functional examination has not been carefully performed for any one of them. In this study, using antibodies specific for Sec22b/ERS-24, it is revealed that endogenous Sec22b/ERS-24 is associated with vesicular structures in both the perinuclear Golgi and peripheral regions. Colabeling experiments for Sec22b/ERS-24 with Golgi mannosidase II, the KDEL receptor, and the envelope glycoprotein G (VSVG) of vesicular stomatitis virus (VSV) en route from the ER to the Golgi under normal, brefeldin A, or nocodazole-treated cells suggest that Sec22b/ERS-24 is enriched in the pre-Golgi intermediate compartment (IC). In a well-established semi-intact cell system that reconstitutes transport from the ER to the Golgi, transport of VSVG is inhibited by antibodies against Sec22b/ERS-24. EGTA is known to inhibit ER–Golgi transport at a stage after vesicle/transport intermediate docking but before the actual fusion event. Antibodies against Sec22b/ERS-24 inhibit ER–Golgi transport only when they are added before the EGTA-sensitive stage. Transport of VSVG accumulated in pre-Golgi IC by incubation at 15°C is also inhibited by Sec22b/ERS-24 antibodies. Morphologically, VSVG is transported from the ER to the Golgi apparatus via vesicular intermediates that scatter in the peripheral as well as the Golgi regions. In the presence of antibodies against Sec22b/ERS-24, VSVG is seen to accumulate in these intermediates, suggesting that Sec22b/ERS-24 functions at the level of the IC in ER–Golgi transport.

Elemental propagation of calcium signals in response-specific patterns determined by environmental stimulus strength

Plant cells can respond qualitatively and quantitatively to a wide range of environmental signals. Ca2+ is used as an intracellular signal for volume regulation in response to external osmotic changes. We show here that the spatiotemporal patterns of hypo-osmotically induced Ca2+ signals vary dramatically with stimulus strength in embryonic cells of the marine alga Fucus. Biphasic or multiphasic Ca2+ signals reflect Ca2+ elevations in distinct cellular domains. These propagate via elemental Ca2+ release in nuclear or peripheral regions that are rich in endoplasmic reticulum. Cell volume regulation specifically requires Ca2+ elevation in apical peripheral regions, whereas an altered cell division rate occurs only in response to stimuli that cause Ca2+ elevation in nuclear regions.

Regional business cycles and the emergence of sheltered economies in the southern periphery of Europe. Bruges European Economic Research (BEER) Papers 7/November 2006

Recent research has highlighted that in the last few years the evolution of regional disparities in many European states has become pro-cyclical. This represents a change with respect to the predominantly anti-cyclical pattern of the 1960s and 1970s. This paper addresses the question of whether and when this change has taken place in the southern periphery of Europe, before analyzing the factors that may have played a role in such a change. The analysis relies on a regional database that includes the evolution of the GDP per capita of NUTS II regions in five European countries (France, Greece, Italy, Portugal, and Spain) between 1980 and 2000. The results of the analysis support the hypothesis of a change towards a pro-cyclical evolution of regional disparities in the cases of Italy, Portugal, and Spain, but not in those of Greece and France. A relationship between these pro-cyclical patters and the emergence of less dynamic sheltered economies is also detected in peripheral regions. This lack of dynamism is related to the fact that numerous peripheral areas in southern Europe have become increasingly dependent on factors such as transfers or public investment and employment, and therefore are less exposed to changes in market conditions.

The R8-BC8 phases and crystal growth in monocrystalline silicon under microindentation with a spherical indenter

The morphology and distribution of high-pressure metastable phases BC8 and R8, formed in monocrystalline silicon under microindentation, were identified and assessed using transmission electron microscopy nanodiffraction analysis. It was discovered that the crystal growth inside the transformation zone was stress-dependent with large crystals in its central region. The crystal size could also be increased using higher maximum indentation loads. The BC8 and R8 phases distributed unevenly across the transformation zone, with BC8 crystals mainly in the center of the zone and smaller R8 fragments in the peripheral regions. Such phase distribution was in agreement with the theoretical residual stress analysis.

Religião, redes sociais e capital social no Município de Rio Grande da Serra na Região do Grande ABC Paulista. Estudo das trocas materiais e simbólicas em contexto de periferia urbana

O município de Rio Grande da Serra está situado em uma região Grande ABC paulista reconhecida nacionalmente por seu desenvolvimento econômico e industrial e pelas lutas políticas e sindicais. Paradoxalmente, se configura, social e territorialmente falando, por uma região de periferia urbana. Resultado da forma como a urbanização, na sociedade moderna, conforma o espaço em regiões centrais e periféricas. Localizado no caminho que ligava Santos à Mogi das Cruzes (século XIX) povoado de Geribatiba decorrente das transformações urbanas ocorridas em toda a região, conquistaria, nos anos 1960, sua autonomia político-administrativa. Nas décadas seguintes testemunhou intenso crescimento populacional, resultado do processo migratório, principalmente de mineiros e nordestinos que tinham as cidades, e indústrias, de São Paulo e Grande ABC como destino. Esse deslocamento de pessoas, e as redes formadas em seu em torno, contribuiu para o desenvolvimento de seu campo religioso. Atualmente, com uma população, em torno, de 46 mil habitantes, possui aproximadamente 180 locais de cerimônias religiosas. Nesse contexto, a tese analisa a inserção regional socioeconômica e religiosa de Rio Grande da Serra, a partir de dados comparativos com os demais municípios, e discute como o regionalismo tem contribuído para seu desenvolvimento econômico. Realiza a caracterização das periferias urbanas, discutindo aspectos que lhes são inerentes, como segregação e vulnerabilidade social. Nesse sentido, a investigação possibilitou a identificação do perfil socioeconômico (renda e escolaridade) dos participantes dos grupos religiosos (católicos, evangélicos, kardecistas e umbandistas), permitindo, também, identificar desigualdades sociais no interior de seu território, constatando que determinados bairros são mais vulneráveis do que outros. Considerando que esse estudo examina a capacidade das redes sociais e religiosas, de aumentar o capital social de seus participantes, foi realizado o mapeamento e etnografia das diversas práticas associativas, mais ou menos formais e estruturadas, de forma a analisar os elementos materiais e simbólicos por elas produzidos. Constatou-se, apoiado na aplicação de questionários, entrevistas e observação participativa, que, a partir do habitus religioso de cada grupo, as redes possibilitam no âmbito econômico questões como emprego e renda ou auxílio em necessidades básicas de sobrevivência, através de campanhas e trabalhos sociais. No âmbito simbólico, as redes propiciam questões importantes à existência humana, como a crença na salvação ou evolução da alma, socialização, autoestima, prestígio ou ainda a expectativa de cura ou tratamento de dependência química. Pôde-se aferir que, a despeito das diferentes formas como cada grupo, e seus participantes, se apropriam do capital social, as redes sociais e religiosas, no município, funcionam como redes de proteção, especialmente à população em situação alta de vulnerabilidade social.

Influence of check and field size on the visual evoked magnetic response to a pattern shift stimulus

A decrease in the check size of a pattern shift stimulus increases the latency and amplitude of the visual evoked potential (VEP) P100. In addition, for a given check size, decreasing the size of the stimulus field increases the latency and amplitude of the P100. These results imply that the central regions of the retina make a significant contribution to the generation of the electrical P100. However, the corresponding magnetic P100m may have a different origin. We have studied the effects of check and field size on the P100m in five normal subjects using a DC-Squid, second-order gradiometer. Magnetic responses were recorded at the positive maximum of the P100m over the occipital scalp to six check sizes (10-100') presented in a large (13 degrees 34') and small (5 degrees 14') field and to a large check (100') presented in seven field sizes (1 degree 45' - 15 degrees 10'). No responses were recorded to any check size with a small field. Decreasing the check size presented in a large field increased latency of the P100m by approx. 30 ms while the amplitude of the response decreased with the largest reduction occurring between 70' and 12' checks. Using a large check, latency increased and amplitude decreased as the field size was reduced. The latency changes in response to check and field size were similar to those described for the VEP although the magnitudes of the magnetic changes were greater. Unlike the VEP, amplitude responses were maximal when large checks were presented in a large stimulus field. This suggests that regions outside the central retina make a more significant contribution to the visual evoked magnetic response than they do to the VEP, and that the P100m may be useful clinically in the study of diseases that affect the more peripheral regions of the retina.

Density and cross-sectional areas of axons in the olfactory tract in control subjects and Alzheimer's disease:an image analysis study

Objective. Using an image analysis system to determine whether there is loss of axons in the olfactory tract (OT) in Alzheimer’s disease (AD). Design. A retrospective neuropathological study. Patients Nine control patients and eight clinically and pathologically verified AD cases. Measurements and Results. There was a reduction in axon density in AD compared with control subjects in the central and peripheral regions of the tract. Axonal loss was mainly of axons with smaller (<2.99 µm2) myelinated cross-sectional areas. Conclusions. The data suggest significant degeneration of axons within the OT involving the smaller sized axons. Loss of axons in the OT is likely to be secondary to pathological changes originating within the parahippocampal gyrus rather than to a pathogen spreading into the brain via the olfactory pathways.

Aszimmetriák az Európai Unión belül (Assimetries within the European Union)

A centrum-periféria kapcsolatok elemzése az Európai Unióra is kiterjeszthető. Az integrációs folyamatban jól elemezhetőek a bourdeiu-i tőkeformák, ezek egymásra történő átváltása, a gazdasági és politikai aszimmetriák kölcsönhatása. E kölcsönös kapcsolatokban továbbra is a gazdasági viszonyok meghatározottsága érvényesül. A világgazdasági válság felszínre hozta a történelmi aszimmetriákat, amelyeket a korábbi neoliberális politikák tovább mélyítettek. Az Unión belüli periferális térségekre a válság különbözőképpen hatott. E hatások semlegesítésére többnyire a megszorító gazdaságpolitikákat alkalmazzák. Továbbra is hiányzik azonban egy, a minőségi s nem mennyiségi szempontokat hangsúlyozó fejlesztési politika. Egy ilyen megközelítés a strukturális, intézményi vonatkozásokat erősítené, s ezzel járulna hozzá a periféria termelési, forgalmi, elosztási képességeinek fokozásához. _____ Analysis of centre-periphery relations can be extended to the European Union as well. The capital forms by Bourdieu, their inter-changeability, mutual effects of economic and political asymmetries could be analysed well in the integration process. In these mutual relations economic relations still play the dominant role. The global economic crisis has brought to the surface historical asymmetries, further aggravated by earlier neo-liberal economic policies. Peripheral regions within the Union have been affected in different ways. In order to neutralise these effects austerity measure have been implemented. However, a development policy, emphasising quality and not only quantitative aspects, is still missing. Such an approach would strengthen structural and institutional elements, further enhancing production, trade, and distribution capabilities of the periphery.

Financing innovative start-ups in Portuguese context: what is the role of business angels networks?

Business angels provide both financing and managerial experience, which increase the likelihood of the survival of innovative start-ups. Over the last years, European countries with developing informal venture capital markets have seen governments support the creation of business angels networks (BANs) to increase and consolidate these markets. Using the Portuguese context to carry out the empirical work, this paper provides an assessment of value added provided by angels’ networks. A total of 88 useable responses were received and analysed using non-parametric statistical techniques. This paper demonstrates that is evidence of positive contribution of BANs in terms of bringing together investors and linking them with entrepreneur’s seeking finance. BANs played an important role in financing innovative start-ups also in peripheral regions. Results lead us to conclude that government support BANs would appear to be an effective mechanism to stimulate the angel market in developing informal venture capital markets. The conclusions of this paper are likely to have relevance for countries where there is growing interest in the potential of business angels as a means of financing innovative start-ups.

INTERMITTENT ACTIVATION OF PERIPHERAL CHEMORECEPTORS IN AWAKE RATS INDUCES Fos EXPRESSION IN ROSTRAL VENTROLATERAL MEDULLA-PROJECTING NEURONS IN THE PARAVENTRICULAR NUCLEUS OF THE HYPOTHALAMUS

Despite the well-established sympathoexcitation evoked by chemoreflex activation, the specific sub-regions of the CNS underlying such sympathetic responses remain to be fully characterized. In the present study we examined the effects of intermittent chemoreflex activation in awake rats on Fos-immunoreactivity (Fos-ir) in various subnuclei of the paraventricular nucleus of the hypothalamus (PVN), as well as in identified neurosecretory preautonomic PVN neurons. In response to intermittent chemoreflex activation, a significant increase in the number of Fos-ir cells was found in autonomic-related PVN subnuclei, including the posterior parvocellular, ventromedial parvocellular and dorsal-cap, but not in the neurosecretory magnocellular-containing lateral magnocellular subnucleus. No changes in Fos-ir following chemoreflex activation were observed in the anterior PVN subnucleus. Experiments combining Fos immunohistochemistry and neuronal tract tracing techniques showed a significant increase in Fos-ir in rostral ventrolateral medulla (RVLM)-projecting (PVN-RVLM), but not in nucleus of solitarii tract (NTS)-projecting PVN neurons. In summary, our results support the involvement of the PVN in the central neuronal circuitry activated in response to chemoreflex activation, and indicate that PVN-RVLM neurons constitute a neuronal substrate contributing to the sympathoexcitatory component of the chemoreflex. Published by Elsevier Ltd on behalf of IBRO.

Potassium channel alterations mediate peripheral nerve hyperexcitability in a mouse model of type 2 diabetes mellitus

Diabetes mellitus (DM) is a major cause of peripheral neuropathy. More than 220 million people worldwide suffer from type 2 DM, which will, in approximately half of them, lead to the development of diabetic peripheral neuropathy. While of significant medical importance, the pathophysiological changes present in DPN are still poorly understood. To get more insight into DPN associated with type 2 DM, we decided to use the rodent model of this form of diabetes, the db/db mice. During the in-vivo conduction velocity studies on these animals, we observed the presence of multiple spiking followed by a single stimulation. This prompted us to evaluate the excitability properties of db/db peripheral nerves. Ex-vivo electrophysiological evaluation revealed a significant increase in the excitability of db/db sciatic nerves. While the shape and kinetics of the compound action potential of db/db nerves were the same as for control nerves, we observed an increase in the after-hyperpolarization phase (AHP) under diabetic conditions. Using pharmacological inhibitors we demonstrated that both the peripheral nerve hyperexcitability (PNH) and the increased AHP were mostly mediated by the decreased activity of Kv1-channels. Importantly, we corroborated these data at the molecular level. We observed a strong reduction of Kv1.2 channel presence in the juxtaparanodal regions of teased fibers in db/db mice as compared to control mice. Quantification of the amount of both Kv1.2 isoforms in DRG neurons and in the endoneurial compartment of peripheral nerve by Western blotting revealed that less mature Kv1.2 was integrated into the axonal membranes at the juxtaparanodes. Our observation that peripheral nerve hyperexcitability present in db/db mice is at least in part a consequence of changes in potassium channel distribution suggests that the same mechanism also mediates PNH in diabetic patients. ∗Current address: Department of Physiology, UCSF, San Francisco, CA, USA.

Altered distribution of juxtaparanodal kv1.2 subunits mediates peripheral nerve hyperexcitability in type 2 diabetes mellitus.

Peripheral nerve hyperexcitability (PNH) is one of the distal peripheral neuropathy phenotypes often present in patients affected by type 2 diabetes mellitus (T2DM). Through in vivo and ex vivo electrophysiological recordings in db/db mice, a model of T2DM, we observed that, in addition to reduced nerve conduction velocity, db/db mice also develop PNH. By using pharmacological inhibitors, we demonstrated that the PNH is mediated by the decreased activity of K(v)1-channels. In agreement with these data, we observed that the diabetic condition led to a reduced presence of the K(v)1.2-subunits in juxtaparanodal regions of peripheral nerves in db/db mice and in nerve biopsies from T2DM patients. Together, these observations indicate that the T2DM condition leads to potassium channel-mediated PNH, thus identifying them as a potential drug target to treat some of the DPN related symptoms.

Characterization of the Morphological and Molecular Changes Associated with Diabetic Peripheral Neuropathy in Type 2 Diabetes

More than 246 million individuals worldwide are affected by diabetes mellitus (DM) and this number is rapidly increasing (http://www.eatlas. idf.org). 90% of all diabetic patients have type 2 DM, which is characterized by insulin resistance and b-cell dysfunction. Even though diabetic peripheral neuropathy (DPN) is the major chronic complication of DM its underlying pathophysiological mechanisms still remain unknown. To get more insight into the DPN associated with type 2 DM, we characterized the rodent model of this form of diabetes, the db/db mice. The progression of pathological changes in db/db mice mimics the ones observed in humans: increase of the body weight, insulin insensitivity, elevated blood glucose level and reduction in nerve conduction velocity (NCV). Decreased NCV, present in many peripheral neuropathies, is usually associated with demyelination of peripheral nerves. However, our detailed analysis of the sciatic nerves of db/db mice exposed for 4 months to hyperglycemia, failed to reveal any signs of demyelination in spite of significantly reduced NCV in these animals. We therefore currently focus our analysis on the structure of Nodes of Ranvier, regions of intense axo-glial interactions, which also play a crucial role in rapid saltatory impulse conduction. In addition we are also evaluating molecular changes in somas of sensory neurons projecting through sciatic nerve, which are localized in the dorsal root ganglia. We hope that the combination of these approaches will shed light on molecular alterations leading to DPN as a consequence of type 2 DM.

Positive and negative regulation of T cell responses by fibroblastic reticular cells within paracortical regions of lymph nodes.

Fibroblastic reticular cells (FRC) form the structural backbone of the T cell rich zones in secondary lymphoid organs (SLO), but also actively influence the adaptive immune response. They provide a guidance path for immigrating T lymphocytes and dendritic cells (DC) and are the main local source of the cytokines CCL19, CCL21, and IL-7, all of which are thought to positively regulate T cell homeostasis and T cell interactions with DC. Recently, FRC in lymph nodes (LN) were also described to negatively regulate T cell responses in two distinct ways. During homeostasis they express and present a range of peripheral tissue antigens, thereby participating in peripheral tolerance induction of self-reactive CD8(+) T cells. During acute inflammation T cells responding to foreign antigens presented on DC very quickly release pro-inflammatory cytokines such as interferon γ. These cytokines are sensed by FRC which transiently produce nitric oxide (NO) gas dampening the proliferation of neighboring T cells in a non-cognate fashion. In summary, we propose a model in which FRC engage in a bidirectional crosstalk with both DC and T cells to increase the efficiency of the T cell response. However, during an acute response, FRC limit excessive expansion and inflammatory activity of antigen-specific T cells. This negative feedback loop may help to maintain tissue integrity and function during rapid organ growth.