Treatment of an Autoimmune Disease with “Classical” T Cell Veto: A Proposal

Immune responses protect against infectious diseases and cancers. In normal circumstances, the immune system is tolerant to self. However, under certain conditions this tolerance is broken. The immune system attacks otherwise normal tissue. An autoimmune disease ensues. Strategies are now being sought that remove the pathogenic T cells without affecting other immune functions. Classical veto has been described as an immune suppressive mechanism able to remove T cells in a highly specific and effective manner. The present article briefly reviews the current knowledge on the development of autoreactive T cells and their regulation in the periphery. It describes classical veto, its mechanisms, and its novel applications. Finally, it argues that classical veto can be adapted to treat an autoimmune disease, such as type I diabetes mellitus.     

Theβ−tubulin genes ofDrosophila auraria are arranged in a cluster

When the 1-, 2- and 3-tubulin-specific DNAs fromDrosophila melanogaster were used as probes to recognize tubulin-specific sequences in the chromosomes ofDrosophila auraria, they were found to hybridize to the same polytene band in region 32C of the 2L polytene chromosome. Three overlapping clones were isolated from a EMBL 3 genomic library ofD. auraria, and they all contain -tubulin-specific sequences based on hybridization and partial-sequencing experiments of subcloned fragments. These clones hybridize in situ to the same polytene chromosome band in region 32C and they represent an approximately 35-kb fragment of genomic DNA.     

Molecular and cytogenetic organization of the 5S ribosomal DNA array in chicken (Gallus gallus)

The 5S ribosomal (r) RNA genes encode a small (120-bp) highly-conserved component of the large ribosomal subunit. The objective of the present research was to study the molecular and cytogenetic organization of the chicken 5S rDNA. A predominant 2.2-kb gene (5S) consisting of a coding and intergenic spacer (IGS) region was identified in ten research and commercial populations. A variant gene repeat of 0.6kb (5S) was observed in some of the populations. Genetic linkage analysis and cytogenetic localization by fluorescence in-situ hybridization assigned the 5S rDNA to chromosome 9. The 5S rDNA array was determined to be 80.2±7.0kb upon electrophoretic sizing following EcoRV digestion. Sequence analysis of 5S IGS regions revealed considerable conservation between chicken subspecies (98.4% identity) as well as homology with vertebrate Pol III promoter and regulatory sequence motifs. Minor intraindividual sequence variation within 1000bp of IGS was observed in four cloned Red Jungle Fowl (Gallus gallus gallus) 5S repeats (95.5% identity in this region). Sequence comparisons between IGS regions of 5S and 5S genes indicated two short continuous (>20bp) and many short non-continuous homologous regions as well as other conserved features such as promoter and termination motifs.     

A simple method for measurement of inulin in nanoliter samples using glass capillaries

Summary A simple method using glass capillaries instead of microcuvettes for measurement of inulin in nanoliter samples is given. Inulin was determined with anthron reagent (5 or 10 nl samples +3 l anthron reagent). Glass capillary tubes (o.d.=1 mm, i.d.=0.68 mm, length=150 mm) in which the chemical reaction took place during incubation at 56°C were directly introduced into the optical system of a Zeiss spectrophotometer PMQ II with sphere attachment and objective.Extinction was measured vertically to the axis of the capillary. The changes of extinction of 20 different capillaries with the blank at different positions was only 1.13×10^–3. The exactness of measurement in the concentration range of 100 200 400 750 1500 3000 mg-% inulin was for 5nl/3 l: 19.8 11.0 6.7 4.7 3.0 2.2%. 10nl/3 l: 13.0 8.4 5.1 3.9%.This method of measurement may also be applicable for other colorimetric reactions with nanoliter samples.This work was supported by Fonds zur Förderung der wissenschaftlichen Forschung.     

Unterschiedlich hoher Ureterdruck in Abhängigkeit von der Diureseform beim Hund

Zusammenfassung Bei Vorliegen einer normalen Diurese wird nach Ureterabklemmung der sog. hohe Ureterdruck, unter osmotischer Diurese der maximale erreicht. Die Differenz von Blutdruck und maximalem Ureterdruck war im Mittel der Versuche um 20 mm Hg kleiner als diejenige des hohen. Die Ursache dafür wird kurz diskutiert.Herrn Prof. Dr.S. Janssen zum 70. Geburtstag gewidmet.     

A comparative immunohistological study of cerebellar enolases

Summary A comparative immunohistological study of the neurone-specific enolase and enolase, demonstrates the exclusive neuronal localization of enolase and its absence from glial cells. In contrast, enolase is located in astroglial cells. The validity of enolase as a neuronal marker and enolase as an astrocytic marker, is confirmed both by a double labelling technique, using antibodies to and to revealed with fluorescence or peroxidase in the same tissue sections, and by immunoelectronmicroscopy.     

A new gene encoding tRNAPro (GGG) is present in the chloroplast genome of black pine: a compilation of 32 tRNA genes from black pine chloroplasts

The chloroplast genome of black pine (Pinus thumbergii), a gymnosperm, contains 32 different tRNA genes, 30 of which correspond to those previously identified in tobacco and rice chloroplast genomes. Two additional genes encode tRNA^Pro (GGG) and tRNA^Arg (CCG); the former is newly identified while the latter is present in liverwort, Physcomitrella patens and Angiopteris lygodiifolia, chloroplast genomes. Moreover, a partial copy of the split tRNA^Gly (UCC) gene and full copies of tRNA^His (GUG), tRNA^Thr (GGU) and tRNA^Ser (GCU) genes are present in the large single-copy region of the genome, suggesting extensive rearrangements of the chloroplast genome during evolutio. No tRNA genes whose tRNA products can recognize codons CUU/C (Leu) and GCU/C (Ala) have been found. We propose that the 32 tRNAs are sufficient to read all the 61 sense codons in the black pine system using the two-out-of-three and the U:N wobble mechanisms.     

In vitro susceptibility ofHaemophilus influenzae to cefaclor,cefixime, cefetamet and loracarbef

The susceptibility of 2,212Haemophilus influenzae isolates cultured in UK clinical laboratories in 1991 was determined for four orally-administered -lactam drugs. These isolates included 1,893 ampicillin-susceptible, 191 -lactamase-positive and 128 ampicillin-resistant, -lactamase-negativeHaemophilus influenzae. While 150 (6.8 %) isolates were resistant to cefaclor (MIC 16 mg/l) and 85 (3.8 %) to loracarbef, all were inhibited by 2 mg/l cefetamet and 1 mg/l cefixime and were therefore susceptible to these agents. Ranges and modes of inhibition zone diameters and MICs indicated that the susceptibility of a variable proportion of the 191 -lactamase-positive isolates to cefaclor, loracarbef and cefetamet was reduced compared with the fully susceptible population. In contrast, a major reduction in susceptibility to all four antimicrobial agents was seen among the 128 ampicillin-resistant (MIC 1–64 mg/l) -lactamase-negative isolates such that these accounted for 53 % and 67 % of the total number of organisms resistant to cefaclor and loracarbef respectively. In addition, 23 of 25 isolates inhibited only by 1 mg/l cefetamet and all eight inhibited only by 0.5 mg/l cefixime showed this type of resistance to ampicillin. Results indicate the importance of detecting non--lactamase-mediated resistance to ampicillin and any concomitant diminished susceptibility to other -lactam drugs.     

Altered synthesis of interferon-γ and expression of interferon-γ receptor by peripheral blood mononuclear cells from patients with IgA nephropathy and non-IgA proliferative glomerulonephritis

Previously we reported disease-specific interaction between interferon- (IFN-) and interleukin-4 (IL-4) in patients with IgA nephropathy (IgAN), suggesting the existence of unusual T cell behavior in this disease. In the present study, we investigated characteristic synthesis of interferon- (IFN-) and expression of IFN- receptor (IFN-R) in the peripheral blood mononuclear cells (PBMC) from patients with IgAN and other chronic proliferative glomerulonephritis (PGN). Heparinized peripheral blood samples were obtained from 38 patients with chronic mesangial proliferative glomerulonephritis (CGN; including 24 with IgA nephropathy) and 20 healthy controls. PBMC were isolated by gradient centrifugation and fragments were cultured in Iscove's modified Dulbecco's medium (IMDM) supplemented with 10% fetal calf serum (FCS) for 72 hr. IFN- concentrations in supernatants were evaluated by the enzyme-linked immunosorbent assay (ELISA). Other parts of PBMC pellets were reacted with anti-human IFN-R monoclonal antibody and FITC-labeled anti-mouse second antibody for analysis of IFN-R expression on these cells by FACScan. The remaining PBMC were fractionated into CD4⁺ T cells, CD8⁺ T cells, B cells, NK, cells and macrophages using the MACS cell sorting system. The isolated cells were evaluated for IFN- or IFN-R mRNA expression by the semiquantitative RT-PCR method.In vitro IFN- synthesis was enhanced in patients with CGN, and NK cells were revealed to be responsible for such enhancement. On the other hand, the expression of IFN-R on macrophages was suppressed in CGN patients. These results suggest that impairment of regulation of the IFN- system might be involved in the development of CGN.     

Lipotropismus von Coxsackie-Virus (Typ B-1) bei ausgewachsenen weißen Mäusen

Zusammenfassung Im braunen Fettgewebe ausgewachsener und hormonal nicht beeinflußter Mäuse findet eine frühzeitige und intensive Vermehrung von Coxsackie-Virus B- 1 (Stamm KLÍ und CONN- 5) statt. Auf die Bedeutung dieses Befundes für den Mechanismus der Coxsackie-Infeklion der weißen Maus wird hingewiesen.     

Die Spulwurmerkrankungen in Darmstadt und Hessen vom Abwasseringenieur gesehen

Ohne ZusammenfassungMit 1 Textabbildung.Im Heft 2 des Gesundheits-Ingenieur 1948 erschien meine Abhandlung Spulwurmplage und Abwasserbeseitigung in Darmstadt. Dem darin behandelten Stoff wurde auch von vielen Hygienikern solches Interesse entgegengebracht, daß ich die Abhandlung in erweiterter Form hier veröffentliche. Die seit der Veröffentlichung im Gesundheits-Ingenieur erzielten Fortschritte sind dabei berücksichtigt.     

Über die „thermo-dilution“-Methode zur Bestimmung des Herzzeitvolumens am narkotisierten und unnarkotisierten Hund

Zusammenfassung 1. Vergleichende Messungen des Herzzeitvolumens mit der thermodilution-Methode, nach dem Fickschen Prinzip und mit dem Farbstoff-Verdünnungs-Verfahren zeigten gute Übereinstimmung und bewiesen die Brauchbarkeit der thermo-dilution-Methode.2. Die Versuche ergaben weiterhin, daß die Herzzeitvolumen-Bestimmung mit der thermo-dilution-Methode auch am unnarkotisierten Tier möglich ist.Mit 4 Textabbildungen     

Supersuppressive “petite” mutants of yeast

Summary A class of suppressive petite mutants of S. cerevisiae, called here supersuppressive, is characterized by a) the fact that their unmodified mitochondrial genomes are the only ones found in the progeny of crosses with wild-type cells; b) very short repeat units (400–900 base pairs) in their mitochondrial genomes. The repeat units of the three supersuppressive petites investigated here share a common 83 nucleotide sequence, which seems to correspond to an initiation site of DNA replication; the multiple copies of this site in the mitochondrial genomes of supersuppressive petites might explain why these genomes can compete out those of wild-type cells.     

Intrinsic burst generation of preinspiratory neurons in the medulla of brainstem-spinal cord preparations isolated from newborn rats

In brainstem-spinal cord preparations isolated from newborn rats, intrinsic burst-generating properties of preinspiratory (Pre-I) neurons in the rostral ventrolateral medulla, which have been suggested to be primary respiratory rhythm-generating neurons, were studied by perforated whole-cell recordings using the antibiotic nystatin. Nystatin causes small pores to be formed in the cells, through which pass small monovalent ions. For blockade of chemical synaptic transmission, perfusate Ca²⁺ concentration was lowered to 0.2 mM and the Mg²⁺ concentration was increased to 5 mM. In Iow-Ca²⁺, high-Mg²⁺ solution (referred to here as low Ca), 10 of 55 Pre-I neurons generated rhythmic bursts (burst type), 14 fired tonically (tonic type), and 31 were silent (silent type). Burst-type neurons showed periodic depolarization of 5–12 mV in low Ca, at a rate of 12±6.5/min. Hyperpolarization of the membrane caused decrease in or disappearance of the periodic depolarization and prolongation of the cycle period. Thus, the burst generations were voltage dependent. The firing frequency of tonictype neurons was 2.3±1.6 Hz and was decreased by hyperpolarization. In 6 of these neurons, the firing patterns changed to burst patterns during continuous hyperpolarization. Membrane depolarization by continuous outward current injection into some silent-type neurons (3 of 11 tested) induced bursting activity. Activity of C4 and Pre-I neurons was completely silent with 0.1–1 M tetrodotoxin (TTX) added to the standard perfusate. In low Ca, burst-type neurons (n=3) were also silent with 1 M TTX perfusion. Inspiratory neurons either became silent (n=4) or fired tonically (n=1) in low Ca. The present study by perforated whole-cell recordings confirmed that some Pre-I neurons possess intrinsic burst-generating properties, which were not attributable to phasic synaptic inputs.     

Control of recombination within and between DNA plasmids of Saccharomyces cerevisiae

Summary [2 m⁺ and [2m°] yeast were transformed to stable leucine prototrophy with the hybrid yeast — E. coli plasmid, pJDB219. This plasmid contains the entire sequence of the endogenous 2 m yeast DNA plasmid in addition to the yeast nuclear LEU2 ⁺ gene and the Co1E1 derivative, pMB9. In the [2 m⁺] transformants, a new wholly yeast LEU2 ⁺ plasmid, pYX, was generated, probably by a recombination event between pJDB219 and 2 m DNA. The plamid, pYX, in the absence of 2 m DNA, was found to exist in equimolar amounts of two forms, A and B, which probably arise by intramolecular recombination across the inverted repeat sequences of the 2 m DNA portion of the plasmid. pJDB219 was found to require the presence of 2 m DNA to undergo this intramolecular recombination. The results suggest that 2, m DNA and pYX code for a gene product required in this recombination event which pJDB219 cannot produce.     

Die Kreislaufanpassung bei Hyperthyreose

Zusammenfassung Es wird über Herzkatheteruntersuchungen an 8 Patienten mit Hyperthyreose berichtet. Zwei Fälle entsprechen in reiner Form der geläufigen Vorstellung des sog. High Output-Zustandes, während die übrigen 6 Fälle verschiedene Varianten der Kreislaufanpassung an die hyperthyreotische Stoffwechsellage darstellen. Die Mechanismen, durch welche solche Anpassungen zustande kommen, werden analysiert. Folgende Reaktionstypen wurden beobachtet: Klassicher High Output ohne Venendrucksteigerung, High Output mit Venendrucksteigerung, relativer High Output mit einseitiger Venendrucksteigerung, normales Herzminutenvolumen bei normalem Venendruck. Auf die Frage der Abgrenzung von Kompensation und Dekompensation des Kreislaufs bei Hyperthyreose wird kurz eingegangen.     

Restricted Vβ gene usage of tumour-infiltrating T lymphocytes in primary gastric malignant B-cell lymphoma

Ten cases of primary gastric malignant lymphoma (PGL) were investigated by immunohistochemical and molecular genetic analysis. These cases were diagnosed histopathologically as follicular small cleaved cell type (1 case), diffuse small cleaved cell type (3 cases) and diffuse large cell type (6 cases) based on the WF (Working Formulation) classification. Seven cases classified as small cleaved or diffuse large cell type belong to low (4 cases) or high (3 cases) grade MALT lymphoma according to Isaacson's classification. All PGL belonged to B lineage cells according to immunohistochemical study and immunoglobulin rearrangements. Rearrangements of TCR chain genes were observed in four of the ten cases. The possibility that the TCR rearrangements were caused by tumour-infiltrating T-cells (TILs) was supported by the following observations: the tumours did not show T- and B-cell biphenotype, TCR exhibited functional VDJ rearrangement and V usage pattern was not a neoplastic type. Analysis of the repertoire of the TCR chain in TILs revealed a common usage of V2 in the above four cases, and furthermore, predominant usage of a particular chain composed of V2-D2.1-J2.3 was observed in one of the four cases. These results indicate that the TILs of PGL have a restricted TCR repertoire.     

Über die Regeneration geschädigter Endothelien nach hartem und weichem Trauma

Zusammenfassung Es bestehen Unterschiede in der Regeneration der Gefäßendothelien nach hartem Trauma (zeitweilige Unterbindung eines Gefäßes durch Seidenfaden) und nach weichem Trauma (zeitweiliges Abklemmen mit einer gummiüberzogenen Klemme), die beschrieben werden. — Am Orte einer früheren Schädigung können Endothelpolypen auftreten, die als Zeichen einer überschießenden Regeneration gewertet werden. — Während nach hartem Trauma ein neues Endothel gebildet wird, das nach Ausreifen einem ungeschädigten Endothel weitgehend ähnlich ist, entstehen nach weichem Trauma mehrkernige Riesenzellen. Diese können mit Zellen alternieren, die nach stärkerer Schädigung schmäler werden und langsam atrophieren, so daß das typische Bild der Regenerationswellen zustande kommt. — Vermutlich können Mikrothromben durch Endothelzellen phagocytiert werden.

---

The regeneration of damaged vascular endothelium after hard and soft traumatism

Summary Differences exist in the regeneration of the vascular endothelium following hard trauma; that is, after temporary ligation of a vessel by silk thread, and following soft trauma, produced by temporary compression with a rubbertipped clamp. The differences are described. Endothelial polyps may appear at the site of previous injury; they are regarded as evidence of an excessive regeneration. With hard trauma a new endothelium is formed which after maturation closely resembles an uninjured endothelium: whereas after a soft trauma multinucleated giant cells develop. The latter may alternate with cells which after more intensive injury become smaller and slowly atrophy, thus giving a typical picture of waves of regeneration. Supposedly microthrombi may be phagocytized by endothelial cells.

---

---

Die Untersuchungen wurden durch Zuwendungen aus dem Wissenschaftlichen Fonds der Gemeinde Wien und aus dem Theodor Körner-Fonds ermöglicht.     

High frequency recombination and the expression of genes cloned on chimeric yeast plasmids: Identification of a fragment of 2-μm circle essential for transformation

Summary We have carried out experiments aimed at explaining the observed variations in transformation frequencies when Saccharomyces cerevisiae or Saccharomyces carlbergensis are transformed with chimeric plasmids that contain one of 4 possible EcoRI fragments of the yeast 2-m circle. These plasmids fall into 2 classes when used to transform 2 different yeast his3 auxotrophs, one (strain LL20) harbours indigenous 2-m circle, and the other (strain YF233) is devoid of this plasmid. Hybrid plasmids containing either the 2.4 mega-dalton (mD) R-form EcoRI fragment (pYF88) or the l.4 mD L-form EcoRI fragment (pYF177) of 2-m circle transform either of the two hosts at a high frequency (50,000 colonies per Mg in LL20 and 10,000 colonies per g in YF233). Hybrid plasmids containing the 1.5 mD R-form EcoRI fragment (pYF87) or the 2.5 mD L-form EcoRI fragment (pYF178) of the 2-m circle transform LL20 at a reduced frequency (6,000–16,000 colonies per g) and YF233 at extremely low frequencies (1–5 colonies per g). All plasmids retrieved from strain YF233 that had been transformed with pYF88 or pYF177 were identical to the original transforming plasmid. Of the plasmids retrieved from strain LL20 that had been transformed with pYF87 and pYF178, approximately half had acquired an extra copy of the 2-m circle. Of the plasmids retrieved from strain LL20 that had been transformed with pYF88 and pYF177, an average of only approximately 13% had acquired an extra copy of 2-m circle. Taken together, these observations indicate that the transformation of yeast by a plasmid lacking the ability to replicate (pYF87 and pYF1780) occurs by the recombinational acquisition of 1 copy of the host 2-m circle, which serves to supply the incoming plasmid with missing essential sequences. A comparison of 2-m circle DNA fragments carried by pYF88 and pYF177 indicates that the region of 2-m circle required for high frequency transformation is a 1.2 mD segment that is common to the 2.4 mD R-form and 1.4 ml) L-form EcoRI fragments. This region extends from the EcoRI cut site adjacent to the PstI site, through to the end of the inverted repeat. However, the inverted repeat sequence alone is not sufficient to bestow high frequency transformation of yeast.     

On the regulation of the expressed L-type calcium channel by cAMP-dependent phosphorylation

The Ca²⁺ channel subunits _1C-a and _1C-b were stably expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293 cells. The peak Ba²⁺ current (I _Ba) of these cells was not affected significantly by internal dialysis with 0.1 mM cAMP-dependent protein kinase inhibitor peptide (mPKI), 25 M cAMP-dependent protein kinase catalytic subunit (PKA), or a combination of 25 M PKA and 1 M okadaic acid. The activity of the _1C-b channel subunit expressed stably in HEK 293 cells was depressed by 1 M H 89 and was not increased by superfusion with 5 M forskolin plus 20 M isobutylmethylxanthine (IBMX). The _1C-a·₂·₂/ complex was transiently expressed in HEK 293 cells; it was inhibited by internal dialysis of the cells with 1 M H 89, but was not affected by internal dialysis with mPKI, PKA or microcystin. Internal dialysis of cells expressing the _1C-a·₂·₂/ channel with 10 M PKA did not induce facilitation after a 150-ms prepulse to +50 mV. The Ca²⁺ current (I _Ca) of cardiac myocytes increased threefold during internal dialysis with 5 M PKA or 25 M microcystin and during external superfusion with 0.1 M isoproterenol or 5 M forskolin plus 50 M IBMX. These results indicate that the L-type Ca²⁺ channel expressed is not modulated by cAMP-dependent phosphorylation to the same extent as in native cardiac myocytes.