Persistence of paradoxical sleep in the cat after destruction of the pontine gagantocellular tegmental field with kainic acid]

The pontine gigantocellular tegmental field (FTG) does not play any role in paradoxical sleep (PS) since the destruction of the cell bodies of the FTG with ka?nic acid does not alter PS in the Cat. Elimination of PS following electrolytic destruction of the ventrolateral pontine reticular formation can be explained by lesions of fibers connecting the region of the locus caeruleus and the bulbar reticular formation.     

Apoptosis and apoptotic mimicry: the Leishmania connection

Different death-styles have been described in unicellular organisms. In most cases they evolve with phenotypic features similar to apoptotic death of animal cells, such as phosphatidylserine (PS) exposure, oligonucleosomal DNA fragmentation, and loss of mitochondrial transmembrane potential, hinting that similar mechanisms operate in both situations. However, the biochemical pathways underlying death in unicellular organisms are still unclear. Host recognition of PS exposed on the surface of unicellular parasites is an important feature of the process of infection and progression of the disease. Here, we discuss data showing that entirely different mechanisms of PS exposure co-exist during the life-cycle of Leishmania amazonensis: in the case of promastigotes, a sub-population dies by apoptosis; in the case of amastigotes, the entire population exposes PS, not necessarily followed by apoptotic death. This phenomenon has been called apoptotic mimicry. The elusive caspase-like activities described in protozoa are also discussed.     

Surface exposure of phosphatidylserine in pathological cells

The asymmetric phospholipid distribution in plasma membranes is normally maintained by energy-dependent lipid transporters that translocate different phospholipids from one monolayer to the other against their respective concentration gradients. When cells are activated, or enter apoptosis, lipid asymmetry can be perturbed by other lipid transporters (scramblases) that shuttle phospholipids non-specifically between the two monolayers. This exposes phosphatidylserine (PS) at the cells outer surface. Since PS promotes blood coagulation, defective scramblase activity upon platelet stimulation causes a bleeding disorder (Scott syndrome). PS exposure also plays a pivotal role in the recognition and removal of apoptotic cells via a PS-recognizing receptor on phagocytic cells. Furthermore, expression of PS at the cell surface can occur in a wide variety of disorders. This review aims at highlighting how PS expression in different cells may complicate a variety of pathological conditions, including those that promote thromboembolic complications or produce aberrations in apoptotic cell removal.Received 26 November 2004; received after revision 3 January 2005; accepted 10 January 2005 Available online 09 March 2005     

Direct inhibition of phospholipid scrambling activity in erythrocytes by potassium ions

The exposure of phosphatidylserine (PS) at the cell surface plays a critical role in blood coagulation and serves as a macrophage recognition moiety for the engulfment of apoptotic cells. Previous observations have shown that a high extracellular [K⁺] and selective K⁺ channel blockers inhibit PS exposure in platelets and erythrocytes. Here we show that the rate of PS exposure in erythrocytes decreases by ~50% when the intracellular [K⁺] increases from 0 to physiological concentrations. Using resealed erythrocyte membranes, we further show that lipid scrambling is inducible by raising the intracellular [Ca²⁺] and that K⁺ ions have a direct inhibitory effect on this process. Lipid scrambling in resealed ghosts occurs in the absence of cell shrinkage and microvesicle formation, processes that are generally attributed to Ca²⁺-induced lipid scrambling in intact erythrocytes. Thus, opening of Ca²⁺-sensitive K⁺ channels causes loss of intracellular K⁺ that results in reduced intrinsic inhibitory effect of these ions on scramblase activity. Received 11 September 2008; received after revision 17 October 2008; accepted 27 October 2008     

Choroid plexus implants rescue Alzheimer’s disease-like pathologies by modulating amyloid-β degradation

The choroid plexuses (CP) release numerous biologically active enzymes and neurotrophic factors, and contain a subpopulation of neural progenitor cells providing the capacity to proliferate and differentiate into other types of cells. These characteristics make CP epithelial cells (CPECs) excellent candidates for cell therapy aiming at restoring brain tissue in neurodegenerative illnesses, including Alzheimer’s disease (AD). In the present study, using in vitro approaches, we demonstrated that CP were able to diminish amyloid-β (Aβ) levels in cell cultures, reducing Aβ-induced neurotoxicity. For in vivo studies, CPECs were transplanted into the brain of the APP/PS1 murine model of AD that exhibits advanced Aβ accumulation and memory impairment. Brain examination after cell implantation revealed a significant reduction in brain Aβ deposits, hyperphosphorylation of tau, and astrocytic reactivity. Remarkably, the transplantation of CPECs was accompanied by a total behavioral recovery in APP/PS1 mice, improving spatial and non-spatial memory. These findings reinforce the neuroprotective potential of CPECs and the use of cell therapies as useful tools in AD.     

Effect of ACTH on the zona reticularis of the rat adrenal cortex: an ultrastructural stereologic study.

The effects of ACTH on the rat adrenal zona reticularis were investigated by stereologic methods. It was found that the zona reticularis cell responsiveness to ACTH is similar to that of the zona fasciculata elements. This excludes that the zona reticularis of the adult rat can only function as the site of destruction of worn-out elements migrating from the adrenal outer zones.     

Activated scramblase and inhibited aminophospholipid translocase cause phosphatidylserine exposure in a distinct platelet fraction

Platelet procoagulant activity is mainly determined by the extent of surface-exposed phosphatidylserine (PS), controlled by the activity of aminophospholipid translocase and phospholipid scramblase. Here, we studied both transport activities in single platelets upon stimulation with various agonists. Besides the formation of procoagulant microparticles, the results show that a distinct fraction of the platelets exposes PS when stimulated. The extent of PS exposure in these platelet fractions was similar to that in platelets challenged with Ca²⁺-ionophore, where all cells exhibit maximal attainable PS exposure. The size of the PS-exposing fraction depends on the agonist and is proportional to the platelet procoagulant activity. Scramblase activity was observed only in the PS-exposing platelet fraction, whereas translocase activity was exclusively detectable in the fraction that did not expose PS. We conclude that, irrespective of the agonist, procoagulant platelets exhibit maximal surface exposure of PS by switching on scramblase and inhibiting translocase activity.Received 8 March 2005; received after revision 19 April 2005; accepted 13 May 2005     

Electroencephalography of the olfactory bulb in relation to prespawn homing

Summary remarks It is evident that the bulbar EEG is a complex response which can be influenced by several methodological and stimulant variables. What EEG patterns tell us about actual behavior remains obscure. The bulbar EEG which is evoked by homestream water is not necessarily a reflection of olfactory memory. The evoked bulbar EEG also does not necessarily demonstrate the salmon has distinguished the home-water from another water in terms of migrational orientation. Despite lack of absolute specificity, a correlation between bulbar EEG and actual behavioral performance has been observed even though some non-home waters evoke responses similar to that of the home water. In general it has been found that the home water response can be distinguished from the response to other natural waters. Failure to obtain complete specificity may be due to a variety of variables which have been alluded to earlier in this section. Thus considering that the evoked EEG is a reflection of the integration of a diverse afferent input, further electrical and computer analyses may eventually permit the decoding of the EEG in terms of behavior. However, if the EEG proves to be more a quantitative rather than qualitative reflection of brain activity, relating EEG to behavior will prove to be of limited value. Since afferent responses in the olfactory bulb are influenced by impulses from higher brain centers, studies on centrifugal aspects during olfactory stimulation may be useful in gaining some qualitative understanding of the home water evoked EEG of the olfactory bulb.     

Reciprocal connections between substantia nigra and medullary reticular formation in the rat

Summary Electrophysiological analysis shows reciprocal connections between substantia nigra and medullary reticular formation (nucleus reticularis giganto celullaris). The nigro-reticular connection appears to be monosynaptic, as shown by antidromic activation, and comprises an ipsi and a contralateral component. Its effect is mainly inhibitory. The reticulonigral component produces mainly excitatory effects and includes fibres from nucleus giganto celullaris and nucleus parvocelullaris in the medullary reticular formation.This work was aided by a grant from CONICIT (projecto 31.26.S1-0412).     

Neurogenic effects of β-amyloid in the choroid plexus epithelial cells in Alzheimer’s disease

β-amyloid (Aβ) can promote neurogenesis, both in vitro and in vivo, by inducing neural progenitor cells to differentiate into neurons. The choroid plexus in Alzheimer’s disease (AD) is burdened with amyloid deposits and hosts neuronal progenitor cells. However, neurogenesis in this brain tissue is not firmly established. To investigate this issue further, we examined the effect of Aβ on the neuronal differentiation of choroid plexus epithelial cells in several experimental models of AD. Here we show that Aβ regulates neurogenesis in vitro in cultured choroid plexus epithelial cells as well as in vivo in the choroid plexus of APP/Ps1 mice. Treatment with oligomeric Aβ increased proliferation and differentiation of neuronal progenitor cells in cultured choroid plexus epithelial cells, but decreased survival of newly born neurons. These Aβ-induced neurogenic effects were also observed in choroid plexus of APP/PS1 mice, and detected also in autopsy tissue from AD patients. Analysis of signaling pathways revealed that pre-treating the choroid plexus epithelial cells with specific inhibitors of TyrK or MAPK diminished Aβ-induced neuronal proliferation. Taken together, our results support a role of Aβ in proliferation and differentiation in the choroid plexus epithelial cells in Alzheimer’s disease.     

ERK activation by Ca2+ ionophores depends on Ca2+ entry in lymphocytes but not in platelets, and does not conduct membrane scrambling

Rapid Ca²⁺-dependent phospholipid (PL) reorganization (scrambling) at the plasma membrane is a mechanism common to hematopoietic cells exposing procoagulant phosphatidylserine (PS). The aim of this research was to determine whether activation of the extracellular signal-regulated kinase (ERK) pathway was required for PL scrambling, based on a single report analyzing both responses induced by Ca²⁺ ionophores in megakaryoblastic HEL cells. Ca²⁺ ionophore-stimulated ERK phosphorylation was induced in platelets without external Ca²⁺, whereas exogenous Ca²⁺ entry was crucial for ERK activation in Jurkat T cells. In both cells, membrane scrambling only occurred following Ca²⁺ entry and was not blocked by inhibiting ERK phosphorylation. Furthermore, ERK proteins are strongly phosphorylated in transformed B lymphoblastic cell lines, which do not expose PS in their resting state. Overall, the data demonstrated that ERK activation and membrane scrambling are independent mechanisms. A. Arachiche, I. Badirou: These authors contributed equally to this work. Received 18 June 2008; received after revision 24 September 2008; accepted 1 October 2008     

CD11b-bearing mononuclear leucocytes and IgA levels in the staging of human immunodeficiency virus infection

Certain immunological parameters (i.e. low CD4+ T cell numbers, high serum soluble CD8) have been described as prognostic factors for the progression of human immunodeficiency virus (HIV) infection to later clinical stages. In the present study we have found in one hundred HIV-infected Spanish patients (81% drug abusers, 7% homosexuals, 6% heterosexuals, and 6% other or unknown risk groups) that CD11b+ peripheral blood mononuclear cells are increased in those with persistent lymphadenopathy as compared to other clinical stages (asymptomatic, AIDS-related complex and AIDS). Serum IgA was significantly increased in AIDS patients, and in patients at any other clinical stage who had concomitant infections (mainly mycobacterial and fungal). CD11b (an integrin with complement receptor functions) may thus be of clinical interest for the staging of HIV-infected patients, and reflect stage-selective immunological changes in mononuclear cell biology during HIV infection. High IgA on the other hand, would be a marker of concomitant infection as well as of disease progression. The results concern mostly drug addicts (the main risk group in Spain), but may apply to the other risk groups because no significant differences were detected between drug addicts (n=81) and non-drug addicts (n=19) for the studied variables (p>0.05).     

A study of DNA synthesis in sea urchin hybrids by the incorporation of H3-thymidine

Zusammenfassung (1) Es wird die DNS-Synthese der letalen SeeigelbastardeParacentrotus ×Arbacia (PA) undParacentrotus ×Sphaerechinus (PS) mit H³-Thymidin untersucht. (2) Dem verschiedenen, der Letalität vorausgehenden Entwicklungstpus (PA ohne, PS mit Chromosomenelimination) entspricht ein verschiedener Verlauf der Hemmung der DNS-Synthese. Parallel dazu wird die Vermehrung in der Anzahl der Kerne verglichen.     

CD11b-bearing mononuclear leucocytes and IgA levels in the staging of human immunodeficiency virus infection.

Certain immunological parameters (i.e. low CD4+ T cell numbers, high serum soluble CD8) have been described as prognostic factors for the progression of human immunodeficiency virus (HIV) infection to later clinical stages. In the present study we have found in one hundred HIV-infected Spanish patients (81% drug abusers, 7% homosexuals, 6% heterosexuals, and 6% other or unknown risk groups) that CD11b+ peripheral blood mononuclear cells are increased in those with persistent lymphadenopathy as compared to other clinical stages (asymptomatic, AIDS-related complex and AIDS). Serum IgA was significantly increased in AIDS patients, and in patients at any other clinical stage who had concomitant infections (mainly mycobacterial and fungal). CD11b (an integrin with complement receptor functions) may thus be of clinical interest for the staging of HIV-infected patients, and reflect stage-selective immunological changes in mononuclear cell biology during HIV infection. High IgA on the other hand, would be a marker of concomitant infection as well as of disease progression. The results concern mostly drug addicts (the main risk group in Spain), but may apply to the other risk groups because no significant differences were detected between drug addicts (n = 81) and non-drug addicts (n = 19) for the studied variables (p greater than 0.05).     

Determination of adhesive rate constant in normal and neoplastic homogeneic cells

Summary The adhesive rate constant (ARC) of neoplastic SGS-2 cells which have a low contact inhibition, is remarkably higher than that of normal homogeneic fibroblasts. This is in contrast with the mutual adhesion theory which states that the loss of contact inhibition is strictly related to the loss of cell recognition and consequently to the loss of cell adhesion capacity.This research was supported in part by the Italian CNR Rome, grant No. 79.00955.04 and in part by the Swiss National Science Foundation, Berne, grant No. 3.499.079. The skilful technical assistance of Mrs Mirjam Valluchi is gratefully acknowledged.     

New observations on the representation of the eye muscle proprioception within the descending trigeminal nucleus.

Units responding to stretch of single extraocular muscles were found in the bulbar and cervical portion of the descending trigeminal nucleus of the lamb. The electrolysis of these bulbar sites provoked degeneration of nervous fibres which could be followed till the first cervical segment of the spinal cord.     

Key role of integrin αIIbβ3 signaling to Syk kinase in tissue factor-induced thrombin generation

The fibrin(ogen) receptor, integrin α(IIb)β(3), has a well-established role in platelet spreading, aggregation and clot retraction. How α(IIb)β(3) contributes to platelet-dependent coagulation is less well resolved. Here, we demonstrate that the potent suppressing effect of clinically used α(IIb)β(3) blockers on tissue factor-induced thrombin generation is linked to diminished platelet Ca(2+) responses and phosphatidylserine (PS) exposure. The same blockers suppress these responses in platelets stimulated with collagen and thrombin receptor agonists, whereas added fibrinogen potentiates these responses. In platelets spreading on fibrinogen, outside-in α(IIb)β(3) signaling similarly enhances thrombin-induced Ca(2+) rises and PS exposure. These responses are reduced in α(IIb)β(3)-deficient platelets from patients with Glanzmann's thrombasthenia. Furthermore, the contribution of α(IIb)β(3) to tissue factor-induced platelet Ca(2+) rises, PS exposure and thrombin generation in plasma are fully dependent on Syk kinase activity. Tyrosine phosphorylation analysis confirms a key role of Syk activation, which is largely but not exclusively dependent on α(IIb)β(3) activation. It is concluded that the majority of tissue factor-induced procoagulant activity of platelets relies on Syk activation and ensuing Ca(2+) signal generation, and furthermore that a considerable part of Syk activation relies on α(IIb)β(3) signaling. These results hence point to a novel role of Syk in integrin-dependent thrombin generation.     

Viscero-somatic reflexes following distension of urinary bladder in cats: role of supraspinal neuraxis

Viscero-somatic reflexes have been studied by recording monosynaptic reflexes following distension of the urinary bladder in intact, decerebrate and spinal animals. It was observed that the viscero-somatic responses following bladder distension are inhibitory in nature and this inhibition was highest in decerebrates and least in spinal animals. The site of viscero-somatic interaction probably lies in the bulbar area (supraspinal) and spinal cord.     

Investigations on the turnover of adrenocortical mitochondria. VII. Effects of ACTH on the half-life of mitochondria from the zona reticularis of the rat adrenal cortex

Summary The half-life of mitochondria from the zona reticularis of the rat adrenal was calculated by determining the radioactivity decay curves of the mitochondrial compartment of 3H-thymidine-injected animals, using autoradiographic methods. ACTH was found to enhance significantly the half-life of the organelles.Acknowledgment. We wish to thank Miss A. Coi and Mr G. Gottardo for the skilled techical assistance. This work was partly supported by a contract with the CNR (No. 74.00226.04/115.3439).     

多孔硅发光中的极化子效应

多孔硅体现了许多新光学性质，本文通过温度依赖的发光，傅立叶红外谱，时间分辨红外谱的观察。发现了些有规律的信息。众所周知，多孔硅在空气中陈化氧化，导致内部纳米尺寸减小。界面层由氢变为氧，我们发现同时伴随着电子态从本征态向极化子态的变化，前者随尺寸减小能量升高，表现为正常的量子限域效应。而后者却随尺寸减小能量降低。表现为量子限域极化子效应。温度依赖的发光谱型和强度变化也清楚地反映了尺寸依赖的极化子行为。因此，我们提出了个基本的物理模型来描述多孔硅中增强的极化子尺寸效应及其光学行为。