A P Nuclear Magnetic Resonance Study of Intracellular pH of Plant Cells Cultivated in Liquid Medium

³¹P nuclear magnetic resonance has been used to study the vacuolar and cytoplasmic pH of Acer pseudoplatanus, Catharanthus roseus, and Glycine max cells grown as cell suspensions. The adaptation of this technique to plant cells grown in liquid medium is described with emphasis on the removal of Mn²⁺ and phosphate from the extracellular medium and on providing the O₂ supply of the cells in the nuclear magnetic resonance tube and the various problems of calibration. Aerobic and anaerobic cells show large differences in their glucose-6-phosphate, their cytoplasmic inorganic phosphate pools, and their cytoplasmic pH. Differences in the relative sizes of the cytoplasmic and vacuolar inorganic phosphate pools have been observed for the three cell strains studied.     

The modelling of a burst-generating neuron with a field-effect transistor analog

An electronic analog that models the burst generating neuronR ₁₅ of theAplysia abdominal ganglion is described. The analog is based on the four branch Hodgkin-Huxley equivalent circuit for a patch of squid axon membrane, with a choice of parameter values appropriate to theAplysia cell membrane. To realize the slow subthreshold oscillations seen in cellR ₁₅ upon exposure to the drug tetrodotoxin (TTX), it was necessary to include two additional conductance branches,g _Na andg _K, to the basic Hodgkin-Huxley circuit. Without these, the analog was capable of generating only action potentials and hence termed the suprathreshold analog. With all six branches operative, bursts very similar to those seen inR ₁₅ were realized, and subsequent inhibition of the Hodgkin-Huxley sodium conductanceg _Na resulted in the desired subthreshold oscillations. The electronic circuitry and the performance of the suprathreshold and complete analog are described. An explanation is offered for the progressive widening of the action potentials within a burst seen inR ₁₅. The analog also simulates the phenomenon of potassium ion accumulation outside the cell membrane during a burst, using a local feedback loop to reduce the potassium equilibrium potential in a manner roughly proportional to the logarithm of the time integral of the outward potassium current. Some consequences of this effect are also discussed.     

Mechanisms involved in parasitic castration: in vitro effects of the trematode Prosorhynchus squamatus on the gametogenesis and the nutrient storage metabolism of the marine bivalve mollusc Mytilus edulis

The mechanisms involved in the parasitic castration of the marine mussel Mytilus edulis by the trematode parasite Prosorhynchus squamatus Odhner, 1905, have been investigated in vitro with two bioassays employing dissociated host tissues. There is no conclusive evidence that P. squamatus affects the secretion of two host neuroendocrine factors, viz., gonial mitosis-stimulating factor and glycogen mobilization hormone, involved in the gametogenesis/nutrient storage cycles of the mussel. In contrast, extracts of P. squamatus sporocysts and cercariae significantly stimulated glycogen mobilization in host glycogen cells and strongly inhibited host gonial mitosis. A gonial mitosis-inhibiting factor (GMIF) was found in the hemolymph of parasitized mussels. The existence of an endogenous GMIF in mantle tissue of uninfected mussels has been demonstrated. This factor appeared to be secreted into the hemolymph during the period of sexual maturity. Whether the parasite acts directly on the host gonia, or by provoking the liberation of this endogenous GMIF, has yet to be ascertained. It would appear, however, that the parasite acts directly on host glycogen cells.     

The origin of the membrane convolute in degranulating platelets. A comparative study of normal and "gray" platelets

Thrombin-stimulated normal platelets contain a membrane system of dilated channels with openings to the exterior. Whether these membranes originate from the surface connected system (SCS), the alpha-granules or internalized portions of the plasmalemma has not yet been defined. The present study traces in series of ultrathin sections the rearrangement of these membranes during shape change, degranulation and internalization of surface membranes in washed normal and "gray" platelets upon the stimulation with thrombin (1 IU/ml). Cationized ferritin (CF) was used as a surface marker in order to recognize internalized portions of the plasmalemma. Within the first seconds after stimulation, both normal and gray platelets changed their shape by extrusion of the SCS membranes. Simultaneously they started to internalize surface membrane and formed surface membrane invaginations closely attached to the outer rim of the cytoskeletal sphere which developed during the internal contraction. CF was internalized in these invaginations. CF was not observed within the system of dilated channels of stimulated platelets, however. Thrombin-stimulated gray platelets showed a markedly reduced number of dilated channels or none at all. This observation may be due to the fact "gray" platelets are deficient in alpha-granules. It is concluded that the dilated system of membranes in degranulated normal platelets originates from membranes of the alpha-granules which have performed compound exocytosis.     

UPTAKE OF GLUCOSE ANALOGUES BY RAT BRAIN CORTEX SLICES: Na+-INDEPENDENT MEMBRANE TRANSPORT

Abstract— The glucose analogues 3-O-methyl-D-glucose and α-methyl-D-glucoside were not metabolized in brain tissue.
The uptake of these two sugars into the intracellular compartment of brain cortex slices was investigated using media with normal and low Na⁺ concentration (replacement of all NaCl with choline Cl). The cellular transport was not Na⁺-dependent. The transport mechanism clearly distinguished between the two sugars in both normal and low Na⁺ media.     

Sudden interruption of leaflet opening by ventricular contractions: a mechanism of mitral regurgitation.

The motion of both mitral cusps and the presence of valvular regurgitation during ventricular contractions were investigated in seven experiments on dogs in which radiopaque markers had been sutured to the cusps and the valve annulus 1-32 wk before the studies. Cineangiograms of the left ventricle were obtained during ventricular ectopic beats, interposed throughout the cardiac cycle (20-99% of cycle length) and during induced variations in the P-R interval (0-200 ms). Mitral regurgitation was observed only during a) weak, early ectopic beats (peak pressure below 34 mmHg) which were incapable of closing the cusps and b) when ventricular contractions suddenly interrupted normal leaflet motion toward the ventricle, during three well-defined periods of diastole (diastolic valve opening, diastolic rebound, and atrial opening). Valve closure following sudden reversal of cusp opening was slow and the leaflets often did not arrive simultaneously at their closed positions. These findings suggest that sudden interruption of leaflet opening by ventricular contractions is an important mechanism of transient mitral regurgitation in the normal heart.     

Veränderungen der Oenozyten und ihre Funktion während der Metamorphose bei Schwärmern und Zahnspinnern

Zusammenfassung VonCerura vinula undSphinx ligustri wurden die Oenozyten im letzten Larvenstadium und in der Puppe untersucht und ihre Struktur beschrieben. Ihre Aktivitätsphasen liegen zur Zeit der beiden Häutungen (Larven- und Puppenhäutung) und im letzten Larveninstar vor der Umfärbung, einem äußerlich sichtbaren Metamorphoseschritt, und vor der Puppenhäutung im Färbungsstadium III. Sie stehen mit den Umwandlungsprozessen, die in den Raupen zu diesem Zeitpunkt stattfinden in deutlichem Zusammenhang. —2–4 Monate nach der Puppenhäutung sind in den diapausierenden Puppen noch aktive larvale Oenozyten vorhanden. — Aktivitätsphasen sind charakterisiert durch viele große und kleine Vakuolen neben kanalartigen Strukturen im Zytoplasma, stark verzweigte Kerne und weitreichende Zellaus- und-einbuchtungen.Im Vorpuppenstadium (Färbungsstadium IV) entstehen die imaginalen Oenozyten aus der Epidermis, sie werden erst kurz vor der Adulthäutung aktiv.Haemozyten, neurosekrethaltig, legen sich dicht an die Oenozyten an und dringen zwischen Zelleinfaltungen ins Innere vor.Lipide, besonders reichlich in aktiven Phasen vorhanden, konnten sowohl im Zytoplasma nachgewiesen werden, als auch ihr Übertritt aus dem Fettgewebe, das den Drüsen eng anliegt.Glykogen tritt ebenfalls in den Oenozyten auf, seine Menge steht aber in keinem merklichen. Zusammenhang mit den Zellrhythmen. Physiologische Versuche beweisen, daß die Oenozyten und auch die Prothorakaldrüsen in aktiven Phasen das Häutungs- und Metamorphosehormon abgeben. Sie lösen beide den Umfärbungsprozeß aus. Gehirne mit neurosekretorischen Zellen in aktiver Phase oder Cholesterin können die Prothorakaldrüsen und z.T. auch die Oenozyten zur Abgabe ihres Hormons anregen.

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Changes of oenocytes and their function during metamorphosis of sphingidae and notodontidae

Summary Oenocytes of the last larval instar and the pupa ofCerura vinula andSpinx ligustri have been examined, and their structure described. The activity phases of the oenocytes at the time of both moultings, as well as during the last larval instar prior to an externally visible color change and prior to the pupal ecdysis i.e. during color change stage III) were clearly related to the process of metamorphosis, which was occurring in the larvae at this time 2–4 months after pupal ecdysis, diapausing pupae still show active larval oenocytes. Activity phases are characterized by many large and small vacuoles in addition to channel-like cytoplasmatic structures, heavily branched nuclei and extensive cell processes and infoldings of the cell membrane. In the pharate pupal stage (colour change stage IV) the imaginal oenocytes originate from the hypodermis, becoming active just prior to the adult ecdysis.Haemocytes containing neurosecretory material attach themselves to the oenocytes and enter through infoldings of the cell membrane. Lipids, which are particularly abundant during active phases, could be demonstrated in the cytoplasm as well as passing from the fatty tissue closely surrounding the glands. Glycogen was also present in the oenocytes. There was, however, no noticeable relation of these materials to the rhythm.Physiological experiments demonstrated that oenocytes as well as prothoracic glands, when active, secrete the moulting and metamorphosis hormone. Both glands initiate the process of colour change. Brain tissue, containing active neurosecretory cells, or cholesterol, may stimulate the prothoracic galnds and the oenocytes to secrete their hormone.

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Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

30 years of repeat expansion disorders: What have we learned and what are the remaining challenges?

Tandem repeats represent one of the most abundant class of variations in human genomes, which are polymorphic by nature and become highly unstable in a length-dependent manner. The expansion of repeat length across generations is a well-established process that results in human disorders mainly affecting the central nervous system. At least 50 disorders associated with expansion loci have been described to date, with half recognized only in the last ten years, as prior methodological difficulties limited their identification. These limitations still apply to the current widely used molecular diagnostic methods (exome or gene panels) and thus result in missed diagnosis detrimental to affected individuals and their families, especially for disorders that are very rare and/or clinically not recognizable. Most of these disorders have been identified through family-driven approaches and many others likely remain to be identified. The recent development of long-read technologies provides a unique opportunity to systematically investigate the contribution of tandem repeats and repeat expansions to the genetic architecture of human disorders. In this review, we summarize the current and most recent knowledge about the genetics of repeat expansion disorders and the diversity of their pathophysiological mechanisms and outline the perspectives of developing personalized treatments in the future.

Tandem repeats represent one of the most abundant class of variations in human genomes, which are polymorphic by nature and become highly unstable in a length-dependent manner. The expansion of repeat length across generations is a well-established process that results in human disorders mainly affecting the central nervous system. At least 50 disorders associated with expansion loci have been described to date, with half recognized only in the last ten years, as prior methodological difficulties limited their identification. These limitations still apply to the current widely used molecular diagnostic methods (exome or gene panels) and thus result in missed diagnosis detrimental to affected individuals and their families, especially for disorders that are very rare and/or clinically not recognizable. Most of these disorders have been identified through family-driven approaches and many others likely remain to be identified. The recent development of long-read technologies provides a unique opportunity to systematically investigate the contribution of tandem repeats and repeat expansions to the genetic architecture of human disorders. In this review, we summarize the current and most recent knowledge about the genetics of repeat expansion disorders and the diversity of their pathophysiological mechanisms and outline the perspectives of developing personalized treatments in the future.     

Role of Interaction and Nucleoside Diphosphate Kinase B in Regulation of the Cystic Fibrosis Transmembrane Conductance Regulator Function by cAMP-Dependent Protein Kinase A

Cystic fibrosis results from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent protein kinase A (PKA) and ATP-regulated chloride channel. Here, we demonstrate that nucleoside diphosphate kinase B (NDPK-B, NM23-H2) forms a functional complex with CFTR. In airway epithelia forskolin/IBMX significantly increases NDPK-B co-localisation with CFTR whereas PKA inhibitors attenuate complex formation. Furthermore, an NDPK-B derived peptide (but not its NDPK-A equivalent) disrupts the NDPK-B/CFTR complex in vitro (19-mers comprising amino acids 36–54 from NDPK-B or NDPK-A). Overlay (Far-Western) and Surface Plasmon Resonance (SPR) analysis both demonstrate that NDPK-B binds CFTR within its first nucleotide binding domain (NBD1, CFTR amino acids 351–727). Analysis of chloride currents reflective of CFTR or outwardly rectifying chloride channels (ORCC, DIDS-sensitive) showed that the 19-mer NDPK-B peptide (but not its NDPK-A equivalent) reduced both chloride conductances. Additionally, the NDPK-B (but not NDPK-A) peptide also attenuated acetylcholine-induced intestinal short circuit currents. In silico analysis of the NBD1/NDPK-B complex reveals an extended interaction surface between the two proteins. This binding zone is also target of the 19-mer NDPK-B peptide, thus confirming its capability to disrupt NDPK-B/CFTR complex. We propose that NDPK-B forms part of the complex that controls chloride currents in epithelia.     

Histologische untersuchungen am wehrsekretbeutel von Cerura vinula L. und Notodonta anceps Goeze (Notodontidae,Lepidoptera)

The larvae of Cerura vinula L. and Notodonta anceps Goeze secrete formic acid for defence. The glandular protective system which forms the acid and changes of the cell structure were studied with the light-microscope.

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Mit dankenswerter Unterstützung der Deutschen Forschungsgemeinschaft.