Overexpression of mouse IsK protein fused to green fluorescent protein induces apoptosis of human astroglioma cells.

Intracellular K(+) plays an important role in controlling ion homeostasis for maintaining cell volume and inhibiting activity of pro-apoptotic enzymes. Cytoplasmic K(+) concentration is regulated by K(+) uptake via Na(+) -K(+) -ATPase and K(+) efflux through K(+) channels in the plasma membrane. The IsK (KCNE1) protein is known to co-assemble with KCNQ1 (KvLQT1) protein to form a K(+) channel underlying the slowly activating delayed rectifier K(+) outward current which delays voltage activation. In order to further study the activity and cellular localization of IsK protein, we constructed a C-terminal fusion of IsK with EGFP (enhanced green fluorescent protein). Expression of the fusion protein appeared as clusters located in the plasma membrane and induced degeneration of both transiently or stably transfected cells.

Chondroitin sulfate A chains enhance platelet derived growth factor-mediated signalling in fibrosarcoma cells.

Platelet derived growth factor is involved in the autocrine growth stimulation of malignant cells, the stimulation of angiogenesis and the recruitment and regulation of tumor fibroblasts. PDGF has been shown to physically interact with glycosaminoglycans which are abundant in the fibrosarcoma cell microenvironment. Aim of the present study was to examine the effects of glycosaminoglycans on the mitogenic function of platelet derived growth factor in two human fibrosarcoma cell lines (B6FS, HT1080). For this purpose exogenously added glycosaminoglycans, regulators of endogenous glycosaminoglycan synthesis (sodium chlorate as selective inhibitor and beta-D-xyloside as a stimulator) and specific glycosidases to cleave cell-associated glycosaminoglycans, were utilized. Platelet derived growth factor demonstrated a growth stimulating effect on B6FS, whereas no effect was evident on HT1080 fibrosarcoma cells. Beta-D-xyloside had no effect on the basal level or the platelet derived growth factor-induced cell proliferation, whereas sodium chlorate severely reduced the basal level of proliferation in both cell lines. Significant co-stimulatory effects of chondroitin sulfate A in combination with platelet derived growth factor BB on the growth of HT1080 and B6FS cells were found. The co-stimulatory effect of chondroitin sulfate A was not due to transcriptional up regulation of platelet derived growth factor receptors genes, but rather to more efficient signalling of tyrosine kinase receptors. In conclusion, this study shows that chondroitin sulfate A can enhance the mitogenic activity of platelet-derived growth factor in fibrosarcoma cells utilizing a pathway which involves tyrosine kinases. This result introduces a new modulating role for chondroitin sulfate in signalling pathways critical for cancer growth.

Tropism of AAV-2 vectors for neurons of the globus pallidus.

A recombinant AAV-2 vector encoding the green fluorescent protein (gfp) under the control of the cytomegalovirus (CMV) promoter was injected into the striatum at varying antero-posterior coordinates. When the virus was delivered to the anterior part of the striatum, transduction efficiency was low and limited to the vicinity of the needle tract. In contrast, after injection into the posterior part of the striatum, in addition to a localized transduced area in the striatum, efficient and widespread transduction was observed at distance from the injection site, in the globus pallidus. In the latter case, labelled cells were also detected in the internal capsule and in the stria terminalis. The number of transduced cells in the striatum increased up to I month and then decreased whereas in the globus pallidus, transduction was maximal as early as 2 weeks post-injection. In the striatum and in the globus pallidus, the labelled cells had a neuron-like morphology. In contrast, in the internal capsule, labelled cells had a glial-like morphology.

Efficient early and sustained transduction of human fetal mesencephalon using adeno-associated virus type 2 vectors.

The success of transplantation of human fetal mesencephalic tissue into the putamen of patients with Parkinson's disease (PD) is still limited by the poor survival of the graft. In animal models of fetal transplantation for PD, antiapoptotic agents, such as growth factors or caspase inhibitors, or agents counteracting oxidative stress enhance the survival and reinnervation potential of the graft. Genetic modification of the transplant could allow a local and continuous delivery of these factors at physiologically relevant doses. The major challenge remains the development of strategies to achieve both early and sustained gene delivery in the absence of vector-mediated toxicity. We recently reported that E14 rat fetal mesencephalon could be efficiently tranduced by adeno-associated virus type 2 (AAV2) vectors and that gene expression was maintained until at least 3 months after transplantation in the adult rat striatum. Here we report that an AAV2 vector can mediate the expression of the EGFP reporter gene under the control of a CMV promoter in organotypic cultures of freshly explanted solid fragments of human fetal mesencephalic tissue as early as 3 days to at least 6 weeks postinfection. These results suggest that AAV2 vectors could be used to genetically modify the human fetal tissue prior to transplantation to Parkinson's patients to promote graft survival and integration.

Olfactory bulb monoamine concentrations vary with time of day.

The olfactory bulb (OB) has been recently identified as a circadian oscillator capable of operating independently of the master circadian pacemaker, the suprachiasmatic nuclei of the hypothalamus. OB oscillations manifest as rhythms in clock genes, electrical activity, and odor sensitivity. Dopamine, norepinephrine, and serotonin have been shown to modulate olfactory information processing by the OB and may be part of the mechanism that underlies diurnal changes in olfactory sensitivity. Rhythmic release of these neurotransmitters could generate OB rhythms in electrical activity and olfactory sensitivity. We hypothesized that these monoamines were rhythmically released in the OB. To test our hypotheses, we examined monoamine levels in the OB, over the course of a day, by high-performance liquid chromatography coupled to electrochemical detection. We observed that dopamine and its metabolite, 3-4-dihydroxyphenylacetic acid, rhythmically fluctuate over the day. In contrast, norepinephrine is arrhythmic. Serotonin and its metabolite hydroxyindoleacetic acid appear to rhythmically fluctuate. Each of these monoamines has been shown to alter OB circuit behavior and influence odor processing. Rhythmic release of serotonin may be a mechanism by which the suprachiasmatic nuclei communicate, indirectly, with the OB.

Burns during Easter festivities in Greece.

Easter is the most important holiday for the Greek Church. It is rich in traditions and rituals but during the Greek Easter festivities, especially at midnight Mass on Easter Saturday night, it is customary to throw fireworks around. These fireworks are not part of the true Easter tradition and they are potentially fatal. Unfortunately, in the past few years, the custom has become more and more popular in Greece. There are some local variations, mainly in the Aegean islands, where homemade rockets are used to have a "rocket war". The rockets consist of wooden sticks loaded with an explosive mixture containing gunpowder and launched from special platforms. Many severe injuries involving loss of sight and limbs as well as major burns are also caused by the use of illegal fireworks at Easter. Every year numerous burn victims are hospitalized. The most affected areas are the face, the upper extremities, and the chest, often in association with slight or severe wounds and injuries. This study presents our department's experience with incidents due to the use of fireworks during Easter festivities.

Pâques est la fête la plus importante pour l'Église grecque. Il est riche en traditions et rituels, mais pendant la fête des Pâques grecques, en particulier à la messe de minuit le samedi de Pâques, il est de coutume de jeter des feux d'artifice autour. Ces feux d'artifice ne font pas partie de la vraie tradition de Pâques et ils sont potentiellement mortels. Malheureusement, au cours des dernières années, la coutume est devenue de plus en plus populaire en Grèce. Il y a quelques variations locales, principalement dans les îles égéennes, où des roquettes artisanales sont utilisées pour faire une «guerre fusée". Les fusées sont constituées de baguettes de bois chargées de substances chimiques contenant de la poudre à canon et lancées en air du sommet de plateformes construites pour l'occasion. Les lésions sévères sont communes comme par exemple la cécité et la perte d'un membre, aussi à cause de l'utilisation de feux d'artifice illégaux à Pâques. Tous les ans nombreuses personnes sont hospitalisées. Les zones corporelles les plus touchées sont le visage, les membres supérieurs et le thorax, souvent en association avec d'autres lésions plus ou moins graves. Cette étude présente l'expérience de notre département des incidents liés à l'utilisation de feux d'artifice lors des fêtes de Pâques.

Immunotherapy of malignant gliomas using autologous and allogeneic tissue cells.

Immunotherapy of brain tumors is rapidly emerging as a potential clinical option [1-3]. The quality and magnitude of immune responses evoked by the new generation anti-tumor vaccines is in general highly dependent on the source or choice of peptide antigens, and as well, a suitable immunopotentiator. Poorly immunogenic antigens, such as those present in tumor cell lysates, may not reliably provide stimulation like recombinant or DNA-encoded protein antigens might be expected to. In addition, the efficacy of the vaccine may depend on inherent counteracting measures of the tumor which dampen immune surveillance and immune effector activity triggered by immunization [4]. Our body has many means of limiting an immune response to our own (self) proteins. In particular, patients with gliomas exhibit a broad suppression of cell-mediated immunity [5-8]. Unfortunately, for most tumor vaccines the induction of local or systemic immune effector cells does not necessarily translate into objective clinical responses or increased survival [9]. Here we review immunotherapeutic approaches against gliomas and recent pre-clinical and clinical initiatives based on cellular or active immunization of the patient's immune system using autologous and allogeneic tissues or cultured cells. Available evidence shows that single modality cancer therapies likely remain suboptimal. Combination regimens targeting the immune system at multiple coordinated levels must be developed, and possibly combined with strategies to inhibit immune suppressive factors if significant clinical benefit is to be achieved.

[Polyacrylic acid cements]. / Kolies tou polyakrylikou oxeos. Synthese--Mechanismos pexes--chreseis.

This paper is a review of the cements of the poly-acrylic acid. The composition and the mechanism of setting of these cements are examined as well as their clinical application.

Calcineurin acts through the CRZ1/TCN1-encoded transcription factor to regulate gene expression in yeast.

Calcineurin is a conserved Ca2+/calmodulin-dependent protein phosphatase that plays a critical role in Ca2+ signaling. We describe new components of a calcineurin-mediated response in yeast, the Ca2+-induced transcriptional activation of FKS2, which encodes a beta-1,3 glucan synthase. A 24-bp region of the FKS2 promoter was defined as sufficient to confer calcineurin-dependent transcriptional induction on a minimal promoter in response to Ca2+ and was named CDRE (for calcineurin-dependent response element). The product of CRZ1 (YNL027w) was identified as an activator of CDRE-driven transcription. Crz1p contains zinc finger motifs and binds specifically to the CDRE. Genetic analysis revealed that crz1Delta mutant cells exhibit several phenotypes similar to those of calcineurin mutants and that overexpression of CRZ1 in calcineurin mutants suppressed these phenotypes. These results suggest that Crz1p functions downstream of calcineurin to effect multiple calcineurin-dependent responses. Moreover, the calcineurin-dependent transcriptional induction of FKS2 in response to Ca2+, alpha-factor, and Na+ was found to require CRZ1. In addition, we found that the calcineurin-dependent transcriptional regulation of PMR2 and PMC1 required CRZ1. However, transcription of PMR2 and PMC1 was activated by only a subset of the treatments that activated FKS2 transcription. Thus, in response to multiple signals, calcineurin acts through the Crz1p transcription factor to differentially regulate the expression of several target genes in yeast.

[Repair of fractured veneers on prosthetic restorations]. / Epidiorthose thrausmenon opseon prosthetikon apokatastaseon.

The repair of an esthetic prosthetic restoration is a problem which has become of interest to the practicing dentist. Many methods have been devised to repair the fractured veneers. This article is referred to one of these methods which uses light-curing resin as a veneering material and silane as a coupling agent. Some of the silane properties and the hydrolisis of silane are mentioned too. Then the mechanism of adhesion to porcelain and to the metal frame are mentioned and in the end the clinical procedure of the repair.

Connecting shock velocities to electron-injection mechanisms.

Electrons can be accelerated by their interaction with nonlinearly saturated electrostatic waves up to speeds with which they can undergo diffusive acceleration across supernova remnant shocks. Here, we model this wave-electron interaction by particle-in-cell and Vlasov simulations. We find that the lifetime of the saturated wave is considerably longer in the Vlasov simulation, due to differences in how these simulation methods approximate the plasma. Electron surfing acceleration which requires a stable saturated wave may thus be more important for electron acceleration at shocks than previously thought. For beam speeds above a critical value, which we estimate here, both simulation codes exclude surfing acceleration due to a rapid wave collapse.

Malabsorption associated with nonmalignant immunoproliferative small intestinal disease.

Two cases of nonmalignant immunoproliferative small intestinal disease (IPSID) presenting with severe malabsorption are described. The first patient had a lymphocytic infiltrate of the lamina propria and lymphoid hyperplasia of the mesenteric lymph nodes and responded to oral tetracycline. The second patient had a polyclonal plasmacytic infiltration of the lamina propria and of the mesenteric nodes and responded only to cytotoxic treatment with cyclophosphamide. These cases represent examples of non-alpha-chain disease benign IPSID.

Cellular contaminants of adeno-associated virus vector stocks can enhance transduction.

Transduction efficiency of different types of recombinant (r)AAV-2 based vectors preparations markedly differed, with apparently no correlation with the replicative titers. Using HeLa cells as target for transduction, 105 and 30 infectious units were necessary to observe one transductant using respectively cesium-chloride-purified rAAV and crude lysates of producer cells obtained by sonication. The purified vectors were however able to transduce HEK-193 cells efficiently, but transgene expression was detected with some delay compared with crude lysates. The unexpected high transduction efficiency of sonicated crude lysates was due to virally mediated gene transfer, since similar sonicated crude lysates, but with no AAV rep and cap genes, did not lead to detection of transgene products after incubation with HeLa cells. Furthermore, sonicated cellular extracts of 293 or 293/T cells given in trans stimulate transduction of HeLa cells by purified rAAV. In contrast, neither extracts from the adenovirus E1-transformed 911 cell line, nor from other cell lines not harboring any adenovirus gene, had enhancing effect on rAAV-mediated transduction. These data suggest that 293 sonicated extracts contain factors which stimulate rAAV-mediated transduction of cells that are normally poorly transduced and offer a system to identify such factors and to characterize further the steps limiting the transfer of gene by AAV vectors.