Untersuchungen über die spezifische und quantitative Erfassung des Liquor-γ-Globulins mit der Zinksulfatfällung

Zusammenfassung Die Frage der quantitativen und elektiven Erfassung des Liquor--Globulins mit der vonRoboz u. Mitarb. beschriebenen Zinksulfatfällung wurde an Modellversuchen mit reinem -Globulin und Albumin und an der elektrophoretischen Auftrennung des mit Zinksulfat aus Liquor präzipitierten Proteins untersucht. Aus reinen -Globulinlösungen wurden in Abhängigkeit zu der in Ansatz gebrachten -Globulinmenge nur 18–46% gefällt. Das Elektrophoresediagramm der mit Zinksulfat gefällten Liquorproteine zeigte, daß neben einer Anreicherung des -Globulins eine Fällung sämtlicher Liquorproteine zustande kam. Die elektrophoretische Auftrennung der im Überstand der Fällung verbliebenen Eiweißkörper ließ neben den übrigen Fraktionen noch eine deutliche -Globulinbande erkennen. Eine elektive und quantitative Fällung des Liquor--Globulins mit der Zinksulfatmethode konnte demnach nicht nachgewiesen werden.Herrn Dozent Dr.H. Bauer danke ich für die Anregung und Unterstützung bei Durchführung dieser Untersuchungen.     

Light-activated ion channels for remote control of neuronal firing

Neurons have ion channels that are directly gated by voltage, ligands and temperature but not by light. Using structure-based design, we have developed a new chemical gate that confers light sensitivity to an ion channel. The gate includes a functional group for selective conjugation to an engineered K(+) channel, a pore blocker and a photoisomerizable azobenzene. Long-wavelength light drives the azobenzene moiety into its extended trans configuration, allowing the blocker to reach the pore. Short-wavelength light generates the shorter cis configuration, retracting the blocker and allowing conduction. Exogenous expression of these channels in rat hippocampal neurons, followed by chemical modification with the photoswitchable gate, enables different wavelengths of light to switch action potential firing on and off. These synthetic photoisomerizable azobenzene-regulated K(+) (SPARK) channels allow rapid, precise and reversible control over neuronal firing, with potential applications for dissecting neural circuits and controlling activity downstream from sites of neural damage or degeneration.     

Specific cytotoxic T lymphocytes in gene therapy

 Cytotoxic T lymphocytes possess the capacity to lyse target cells which express antigens on their surface recognized by the T cell receptor. These cells are crucial in the body’s defense against foreign antigens. It has long been a goal of tumor biology to utilize T cells, specialized in the elimination of unwanted cells, for the treatment of cancer. The killing activity of T lymphocytes is restricted to specific antigen-presenting cells. For this reason the use of cytotoxic T cells in the elimination of cancer cells is limited to cancer cells which present neoantigens on their surface. To circumvent this limitation we describe a procedure in which the ζ component of the T cell receptor is genetically manipulated and equipped with an extracellular recognition domain. Introduction of a chimeric gene, consisting of the ζ chain of the T cell receptor and a single-chain antibody domain, into cytotoxic T lymphocytes results in T cells with a predetermined recognition specificity for particular tumor cells. The MHC restriction of target cell recognition can be avoided and tumor cells recognized by the single chain antibody domain can be recognized and lysed. Retroviral-mediated gene transduction was used to introduce chimeric ζ chain constructs into primary T cells of mice. The cocultivation of retrovirus producing helper cells with in vitro activated T lymphocytes led to a high gene transduction efficiency into primary T cells. These primary T cells assumed a predetermined specificity for target cell recognition and lysis. The production and provision of tumor cell specific T lymphocytes might not be sufficient to eradicate large tumors in vivo. Using a Schwannoma cell line, we showed that transplanted tumors secrete transforming growth factor β and thereby stifle the action of lymphocytes. We suggest that a coordinated strategy including the suppression of tumor cells specific antilymphocyte action and the provision of tumor cell specific T cells might be required to successfully eliminate tumor cells in vivo. Received: 13 September 1996 / Accepted: 30 October 1996     

Separate and variably shaped chromosome arm domains are disclosed by chromosome arm painting in human cell nuclei

Fluorescence in situ hybridization (FISH) with microdissection probes from human chromosomes 3 and 6 was applied to visualize arm and subregional band domains in human amniotic fluid cell nuclei. Confocal laser scanning microscopy and quantitative three-dimensional image analysis showed a pronounced variability of p- and q-arm domain arrangements and shapes. Apparent intermingling of neighbouring arm domains was limited to the domain surface. Three-dimensional distance measurements with pter and qter probes supported a high variability of chromosome territory folding.     

Aerobic power and pubertal peak height velocity in Belgian boys

Summary To determine the cardiorespiratory response to maximal exercise before, during and after the pubescent growth spurt, thirty boys were tested at yearly intervals over a period of six consecutive years. For each individual, peak height velocity (PHV) was determined. The age at PHV (¯X= 13.6 years) was taken as a standard of maturation. The results from all subjects at 1.5 and 0.5 years before and 0.5 and 1.5 years after PHV are presented. The highest oxygen uptake ( ) obtained during an incremental bicycle ergometer test to voluntary exhaustion was taken as peak oxygen uptake ( peak). Across each of the four years studied, mean peak (min=49.6; max=52.5 ml·kg^–1·min^–1) and mean heart rate (HR) at peak (min=190; max=192) did not change significantly as a function of PHV. On the other hand, the respiratory quotient at peak increased considerably from mean minima and maxima of 0.99 and 1.01 before PHV to 1.07 and 1.10 after PHV. Ventilatory equivalent for ( ), taken as an indicator of ventilatory economy, seemed to be unaffected by the maturation process. The steepest increase in circumpubertal oxygen pulse was found one year after PHV. Average stability coefficients (¯r), calculated from the inter-years correlations were high for height (¯r=0.95), weight (¯r=0.92), HR at peak (¯r=0.74), peak in 1/min (¯r=0.71), oxygen pulse (¯r=0.68) and tidal volume (¯r=0.64).     

The time course of glycogen depletion in single fibers of chronically stimulated rabbit fast-twitch muscle

A time course study was conducted to investigate the possibility of a relationship between fiber degeneration and glycogen depletion in chronically nerve-stimulated extensor digitorum longus muscle of the rabbit. Muscles were stimulated 12 h daily at 10 Hz using alternating one-hour periods of stimulation and rest. When measured for the first time after 3 h (1 h stimulation, 1 h rest, 1 h stimulation), microphotometry revealed complete glycogen depletion of all fiber types (fast glycolytic, FG; fast oxidative glycolytic, FOG; slow oxidative, SO). Different responses were noted beginning at day 4. At this time point, all FOG and SO fibers recovered their glycogen stores with some of the FOG population attaining levels higher than the FOG fibers in the unstimulated, contralateral muscle. Approximately 28% of the FG fibers recovered to normal glycogen values, whereas 58% remained depleted and 14% displayed overshoting glycogen levels. Fifteen percent of all fibers were glycogen-depleted after 12 days of stimulation. At this time, classic fiber types could no longer be distinguished. Fiber degeneration, which was recognized by the invasion of nonmuscle cells, began after 6 days and was restricted to the glycogen-depleted fibers. By this time, there was also a significant increase in DNA content. Exhaustions of glycogen, the main fuel of the FG fibers, is believed to cause a collapse of energy-supply and ATP-driven ionic pumps. The latter could be the initial step of fiber deterioration.     

Immunohistochemical localization of neurofilaments and neuron-specific enolase in 29 cases of neuroblastoma.

Twenty-nine neuroblastomas have been examined with the use of rabbit antibodies specific for each of the three neurofilament polypeptides, with a monoclonal antibody specific for the NF-L polypeptide, and with a rabbit antibody specific for neuron-specific enolase. When frozen material was used, all neuroblastomas were positive with the neurofilaments antibodies. When alcohol-fixed paraffin-embedded material was used, neurofilament staining was weaker and the fixation procedure appeared to destroy the epitopes recognized by the NF-L antibodies preferentially. Although all neuroblastomas were positive for neurone-specific enolase, so were two rhabdomyosarcomas, suggesting that NSE is not an appropriate marker to distinguish the different small blue cell tumors of children.