Dual Vα T cells

The assumption that T cells can only express a single receptor for antigen has in recent years been shown to be incorrect. However, the finding that a substantial number of T cells express two distinct antigen receptors at the cell surface raises a number of questions. In particular, it has been suggested that cells expressing low levels of a self-reactive T cell receptor may escape self-tolerance mechanisms and in certain situations trigger the onset of autoimmune disease. Such a hypothesis in turn raises questions central to the understanding of the nature of T cell recognition and the process of thymocyte maturation.     

17β-Estradiol-sensitivity of cultured myometrial cells

Summary The estrogen sensitivity of cells cultured from the rat myometrium was studied by growing the cells in the absence or presence of 1 nM 17-estradiol. Following a time lag of 10 days, exposure to estrogen resulted in increased incorporation of radiothymidine by the cells. Estrogen treatment also decreased isoproterenol-dependent and GTP-dependent adenylate cyclase activity, but had no effect on basal activity. These cultured cells have been shown previously to have some properties of uterine smooth muscle. The effects estrogen has in viyro, therefore, may reflect important properties in vivo that account for the mechanism by which the sex steroid decreases the sensitivity of the myometrium to isoproterenol.     

β-phenylethyl isothiocyanate-mediated apoptosis in hepatoma HepG2 cells

-Phenylethyl isothiocyanate (PEITC) is a promising chemoprotective compound that is routinely consumed in the diet as its glucosinolate precursor. Previous studies have shown that PEITC can inhibit phase I enzymes and induce phase II detoxification enzymes along with apoptosis in vitro. The detailed mechanisms involved in the apoptotic cascade, however, have not been elucidated. In the present study, we demonstrate that PEITC can induce apoptosis in hepatoma HepG2 cells in a concentration- and time-dependant manner as determined by TUNEL positive and SubG1 population analysis. Caspase-3-like activity and poly(ADP-ribosyl)polymerase cleavage increased during treatment with 20 µM PEITC; high concentrations, however, induced necrosis. Pre-treatment with Z-VAD-FMK and the caspase-3-specific inhibitor Ac-DEVD-CHO prevented PEITC-induced apoptosis, as determined by caspase-3-like activity and DNA fragmentation. Additional investigations also showed that at concentrations of 5-C10 µM PEITC, DNA synthesis was inhibited and G2/M phase cell cycle arrest occurred, correlating with an alteration in cyclin B1 and p34^cdc2 protein levels. Furthermore, we also demonstrate a concentration- and time-dependant burst of superoxide (O₂^-) in PEITC-treated cells. However, pre- and co-treatment with the free radical scavengers Trolox, ascorbate, mannitol, uric acid and the superoxide mimetic manganese (III) tetrakis (N-methyl-2-pyridyl) porphyrin failed to prevent PEITC-mediated apoptosis. Taken together, these results suggest that PEITC potently induces apoptosis and cell cycle arrest in HepG2 cells and that the generation of reactive oxygen species appears to be a secondary effect.Received 23 December 2002; accepted 22 April 2003     

Nestin expression – a property of multi-lineage progenitor cells?

Tissue-specific progenitor cells are characterized by proliferation and differentiation, but, in contrast to embryonic stem (ES) cells, have limited capacities for self-renewal and no tumourigenic potential. These latter traits make progenitor cells an ideal source for regenerative cell therapies. In this review, we describe what is currently known about nestin, an intermediate filament first identified in neuroepithelial stem cells. During embryogenesis, nestin is expressed in migrating and proliferating cells, whereas in adult tissues, nestin is mainly restricted to areas of regeneration. We show that nestin is abundant in ES-derived progenitor cells that have the potential to develop into neuroectodermal, endodermal and mesodermal lineages. Although it remains unclear what factors regulate in vitro and in vivo expression of nestin, we conclude that nestin represents a characteristic marker of multi-lineage progenitor cells and suggest that its presence in cells may indicate multi-potentiality and regenerative potential.     

Ammonia photoproduction byCyanospira rippkae cells ‘entrapped’ in dialysis tube

Summary The photoproduction of ammonia by cells of the heterocystous cyanobacteriumCyanospira rippkae in the presence of the glutamine synthetase inhibitor, L-methionine D,L-sulfoximine (MSX), was investigated. The time course of changes in protein, pigment and carbohydrate concentrations and the C₂H₂-reducing activity of nitrogenase in MSX treated and untreated filament suspensions was also determined. The results show that nearly 40 h after MSX addition the cells are able to recover from the nitrogen starvation induced by the inhibitor by themselves, without the removal of MSX or the addition of nitrogenous compounds. Biliproteins, mobilized as a consequence of MSX addition, seem to play a key role in the process of cell recovery. These findings were exploited in a semicontinuous ammonia producing process with cells immobilized in a dialysis tube photobioreactor.     

The rate of cleavage ofβ-mercaptopyruvate by rapidly dividing cells

Zusammenfassung Quantitative Analysen des-mercaptobrenztraubensäure-spaltenden Enzyms zeigen, dass Zellen, die sich in aktivem mitotischem Prozess befinden (bzw. Krebszellen und Gewebekulturen), dieses Enzym in viel geringerem Masse enthalten als « normale » Zellen. Da dieses Enzym in dem anaeroben Stoffwechsel von Zystein eine wichtige Rolle spielt, lässt sich vermuten, dass sich der Zysteinstoffwechsel der Tumorzelle quantitativ von dem der Normalzelle unterscheidet.

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Supported by U.S. Public Health Grants H-2897, C-3211 and by a grant of the American Cancer Society (70-4612-24).     

The classical ‘Golgi apparatus’ in the vertebrate nerve cells

Zusammenfassung Der Golgi-Apparat in den Neuronen des Rückenmarks der Wirbeltiere entsteht aus Ablagerungen von Osmium oder Silber an Nissl-Körperchen und an mit ihnen verbundenen Canaliculi und Spaltsystemen. Lipoidkörperchen und/oder Mitochondrien sind in bezug auf Bildung von sekundärer Bedeutung.     

Epidermal growth factor stimulates proliferation of rat hepatoma cells producing α-fetoprotein

Summary Epidermal growth factor stimulated both [³H]thymidine uptake and proliferation of rat AH66 hepatoma cells. However, the increase in cell number was not accompanied by a proportional increase in the levels of -fetoprotein of the culture media. The effects of EGF on the cell proliferation were antagonized by N⁶, O²-dibutyryl cAMP.     

‘Chloride-cell’ — like mitochondria-rich cells of salamander larva gill epithelium

Summary Two types of mitochondria-rich cells (MRC) are described ultrastructurally in the gill epithelium of salamander larva. They resemble MRC found in larval ventral epidermis. Histochemical localization of carbonic anhydrase indicated numerous positive reacting cells, most of them flask-shaped. Morphological and functional similarities to fish chloride cells are discussed.     

Protein turnover in glycine-C14-labelled Landschütz ascites tumor cells

Zusammenfassung Untersuchungen über die Stabilität der Eiweißstoffe von Landschütz-Aszites-Tumoren wurden an C¹⁴-Glykokoll,-Valin oder -Leucin markierten Zellen durchgeführt. Es wurde festgestellt, dass während der Zellteilung und des Wachstums kontinuierlich etwa 5–10% pro Tag der einmal gebundenen C¹⁴-Aktivität als niedrigmolekulare Komponenten in die Aszitesflüssigkeit ausgeschieden wird. Diese Instabilität kommt offenbar durch intrazellulare Umsetzung der Eiweißstoffe und nicht durch Ausscheidung ganzer Eiweissmoleküle zustande.     

Occurrence of ‘partially acid-fast’ cells in cultures of genusStaphylococcus and genusMicrococcus

Résumé Lorsqu'on applique aux populations des genresStaphylococcus etMicrococcus une coloration semblable à celle deMachiavello, on obtient des cellules différemment colorées.Ce caractère « partiellement acido-résistant » diffère du caractère tinctorial de l'acido-résistance vraie.

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These studies were done with the aid of the Medical Research Council of Canada. Grant MA-729, 1962.     

Does an initial phasic response exist in the receptor potential of taste cells?

The depolarizing receptor potentials to 0.5 M NaCl recorded from frog taste cells did not exhibit any phasic response, even when the rectangular waveform of stimulus onset was employed. The quickest depolarizations recorded reached the peak in 50 msec. On the other hand, the gustatory neural response showed initial overshoot of the impulse discharge even when 0.5 M NaCl was delivered at the slower rate of 0.06 ml/sec. It is concluded that the initial neural response may be associated with the rate of rise of the receptor potential before its plateau level is reached.     

β-Adrenergic stimulation of androgen production by fetal mouse Leydig cells in primary culture

Summary The responsiveness of fetal mouse Leydig cells to catecholamines (epinephrine, norepinephrine), a-agonist agent (L-isoproterenol) and hCG was investigated in vitro. Fetal Leydig cells when freshly isolated were unable to respond to L-isoproterenol (10^–5M). However, L-isoproterenol, epinephrine and norepinephrine significantly stimulated androgen production by fetal Leydig cells after 24 h of primary culture. Androgen production was increased in both conditions and to a greater extent by hCG. Propranolol blocked the stimulatory effect of L-isoproterenol and epinephrine. It is concluded that catecholamines can regulate fetal testosterone biosynthesis.     

What constitutes a network in the acinar cells of amphibian and mammalian pancreas?

Résumé Il a été montré que l'appareil réticulaire classique de Golgi dans les cellules pancréatiques des amphibies et des mammifères est un produit artificiel. Un certain nombre de raisons sont proposées pour expliquer ce fait.     

Interferon-β can induce the production of plasminogen activator by cultured human cancer cells

Summary Three cultured human cell lines, renal cancer cells (ACHN), bladder cancer cells (EJ), and fibroblasts transformed in culture by Co-60 gamma rays (KMST-6), when treated with interferon-, produced 1.5 to 4 times as much plasminogen activator as the untreated control cultures. This enhanced production of PA was inhibited by cycloheximide or actinomycin D.     

Protein kinase C-dependent NF-κB activation is altered in T cells by chronic stress

Chronic stress has been associated with impaired immune function. In this work we studied the effect of chronic mild stress (CMS) exposure on the early intracellular pathways involved in T cells after stimulation with mitogen. We found that mitogen stimulation of T lymphocytes from CMS-exposed mice resulted in a reduction of the intracellular [Ca²⁺] rise, an impairment of growth-promoting protein kinase C (PKC) activation, a lower NF-κB activation and an increase in the inhibitory cAMP-protein kinase A (PKA) pathway activity with respect to those found in control lymphocytes. However, T cell activation with the direct PKC activator phorbol 12-myristate 13-acetate plus calcium ionophore led to a similar proliferative response in both CMS and control lymphocytes, indicating that signals downstream of PKC would not be affected by stress. In summary, our results show that chronic stress induced an alteration in T cell early transduction signals that result in an impairment of the proliferative response.Received 11 February 2005; received after revision 20 May 2005; accepted 6 June 2005     

A postsynaptic α, 2-receptor: The alpha-adrenergic receptor of hamster white fat cells

Summary The effects of selected -agonists and -antagonists on theophylline-induced lipolysis were investigate in isolated hamster white fat cells ₂-Agonists (tramazoline, clonidine) inhibited theophylline-induced lipolysis while an ₂-agonist (methoxamine) was without any effect. The inhibitory effect of ₂-agonists was suppressed by yohimbine (₂-antagonist), whereas ₂-antagonists were inefficient. This result implies that the -adrenergic receptor of hamster fat cells is of the ₂-type, although located postsynaptically.Acknowledgments. This work was supported by grants from CNRS (ERA 412) and DGRST (grant No. 787 1078). We thank M. Dauzats for excellent technical assistance. We thank Prof. H. Schmitt for tramazoline and AR-C 239 and for helpful discussion.     

Effect of radical scavengers on TNFα-mediated activation of the uPA in cultured cells

Active oxygen, produced by cultured cells following stimulation with various growth factors, seems to be involved in signal transduction leading to cellular responses such as gene expression and growth modulation. In the present study, the intracellular oxidation state was measured in immortalized human endothelial cells (ECV304) after treatment with tumor necrosis factor (TNF), using a fluorescent dye and a laser-scanning confocal microscope. The intracellular oxidation state was increased 60 min after the addition of TNF, and this increase was abolished by a radical scavenger, N-acetylcysteine (NAC), which is also a precursor of glutathione, and by pyrrolidine dithiocarbamate (PDTC). TNF increased the steady state level of urokinase-type plasminogen activator (uPA), and NAC inhibited this increase at a dose that also inhibited the increase in the intracellular oxidation state. PDTC, on the other hand, did not affect the induction of the uPA gene by TNF. These results suggest that intracellular glutathione level rather than the oxidation state is necessary for the induction of the uPA gene by TNF.     

Bacillus thuringiensis δ-endotoxin: Evidence that toxin acts at the surface of susceptible cells

Summary Enzymically activated -endotoxin ofBacillus thuringiensis covalently bound to Sephadex beads, has the same effect on insect cells in tissue culture as free toxin. The effect is prevented by antitoxin antibody and heat denaturation and is not due to a nonspecific protein effect, the beads, or toxin released from the beads. The toxin, therefore, probably acts at the cell surface.