Key word: Chromosome

The word chromosome has survived for over 100 years, because it succinctly defines what early cytologists were able to see with the most modern instrument of their time, a light microscope. It was introduced in a review that became widely known and was published almost simultaneously in German, English and French (Waldeyer 1888, 1889, 1890a, 1890b). In the late 19th century, these three languages were in strong competition for international status as the idiom of science. At the same time, Greek was also considered as a candidate for a nationalistically neutral language of science, and it seems more than coincidence that the word ó matches well the coherent Greek terminology used to describe the cell cycle in mitosis as well as meiosis. Emil Heitz (1935) maintained – in the face of reactionary German efforts to replace the term – that in using the ineradicable word chromosome we think last of all that it indicates a body that stains intensely. Significantly, the key word is no longer restricted to eukaryotes, but has been readily adopted by microbial geneticists (Heidelberg et al. 2000) and acknowledged as defining the elementary unit of genomic partition.     

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.     

Activation of atrial muscarinic K+ channels by low concentrations ofβγ subunits of rat brain G protein

Effects of G protein subunits from rat brain on cardiac K⁺ channel was examined in single atrial cells of guinea-pig, using patch clamp techniques. We found that 10 pM concentration of rat brain subunits preparation could activate the atrial muscarine receptor-gated K⁺ channel (I_K.ACh). Neither the detergent, CHAPS, used to suspend nor the boiled preparation activated I_K.ACh. Furthermore, preincubation of subunits preparation in Mg²⁺-free solution, which easily inactivated -GTP-S, did not affect -activation of I_K.ACh. We concluded, therefore, that subunits themselves can activate I_K.ACh.Supported by the grants from the Ministry of Education, Culture and Science of Japan and from the Calcium Signal Workshop on Cardiovascular Systems     

Immunohistochemical studies on S-100 cells in the anterior pituitary gland of Sprague Dawley rats and spontaneous dwarf rats

The S-100 cells in the pituitary glands of adult male Sprague Dawley rats (SDs) and spontaneous dwarf rats (SDRs) were immunohistochemically examined using anti-S-100 and anti-S-100 monoclonal antibodies. The immunoreactive cells against S-100 protein were divided into three subtypes on the basis of their immunore-activity against subunits of S-100 protein: S-100 dominant type (the -type cell), S-100 dominant type (the \-type cell) and immunoreactive against both S-100 and S-100 (the -type cell). In the SD, -type cells represented 26% of the total S-100 immunoreactive cells (S-100 cells) and were localized in the peripheral area of the ventral region of the pituitary gland. This type of cell was observed forming clusters, with more abundant cytoplasm than the -type cell. The proportion of -type cells was 53%. They were diffusely distributed throughout the gland, and their processes were thicker than those of the -type cell. In the SDR, the proportion of -type cells was 55%, and they were observed throughout the gland. In contrast, -type cells totalled 12% and were localized in small areas of the central and peripheral region of the gland. The proportion of -type cells was 21% in the SD and 33% in the SDR and they were observed forming small clusters in both animal groups. The proportion of -type cells compared with the total of S-100-immunoreactive cells was significantly higher (P < 0.05) in the SDR than in the SD, while the proportion of -type cells was markedly lower (P < 0.05).     

The effects of iron and temperature on the growth ofListeria monocytogenes in cell-free media

The effects of variable iron and temperature on the growth ofL. monocytogenes in vitro was investigated in cell-free tissue culture Medium 199. Two temperatures, 37 and 42C, and varying concentrations of iron (Fe⁺³) derived from ferric ammonium citrate, 40–130g/ml, were selected for the study. Under these conditions,L. monocytogenes exhibited better growth at 37C than at 42C. Dose-effect was demonstrated at every succeeding higher Fe⁺³ concentration except 130g/mI which was inhibitory. The optimal Fe⁺³ concentration at both temperatures was 100g/ml. The implications of these results for tissue culture models and host-parasite relationship, are discussed.     

Effect of theophylline onβ-adrenergic receptor density and cAMP content in bovine aortic smooth muscle cells

Activation of vascular-adrenergic receptors prevents an increase in vascular permeability caused by free radicals or inflammatory peptides. Methylxanthines seem to have similar protective effects on vascular endothelium. In the present study we investigated the effect of theophylline on the-adrenergic receptor expression and cAMP concentrations in cultured endothelial and smooth muscle cells from bovine aorta. Comparable values for-receptor density and binding affinity were detected in both cell types. Isoproterenol induced significant downregulation of-receptors in endothelial (BAEC: –60.5%) and smooth muscle cells (BASMC: –52.5%; P < 0.01). Incubation of endothelial cells with theophylline (4 µg/ml and 40 µg/ml) for 24 hours did not affect-receptor expression, whereas in smooth muscle cells the-receptor density was reduced for –31.5% and –28.7, respectively. In endothelial cells a transient effect on cAMP concentrations was observed after stimulation with isoproterenol (1 µM), but no effect was found in theophylline treated endothelial cells. Stimulation of intact smooth muscle cells with isoproterenol and theophylline (4 µg/ml and 40 µg/ml) resulted in a significant increase of cAMP concentrations after 60 and 240 minutes. The present data suggest a novel, celltype specific effect of theophylline on the-adrenergic receptor expression in vascular smooth muscle cells in vitro.     

Comparison of the estrous cycles in mice of high and low cancer lines

Summary In February and May the author studied the duration of the estrous cycle stages in female mice of high A and low C₅₇ cancer lines. It was established that the average duration of the estrus is much greater and of diestrus much shorter in the A than that of the corresponding stages in the C₅₇ line.The estrous cycles of the C₅₇ line females do not undergo any significant seasonal changes while in the A line they approach the character of the normal mice spring cycles.Desviations from the normal duration of the estrous cycle stages in the high cancer line mice denote a disturbed hypophyseal — ovarian hormonal balance.Presented by Active Member AMN SSSR V. N. Chernigovskii     

Effect of lysolecithin on blood vessel reactivity and of some ethers and esters of glycerophosphocholine and phosphocholine,respectively, on aggregation of rat platelets

Infusion of lysolecithin (LPC; e.g. 88 g/ml for 0.5–1.0 min) did not significantly impair the vasopressor action of norepinephrine (NE), prostaglandin F₂ (PGF₂) and extract of posterior pituitary (EPP) in the isolated perfused hind legs of rats. In other words, vascular smooth muscle behaves differently from the smooth muscle of the guinea-pig small intestine, since, in the latter, contractions evoked by acetylcholine, prostaglandins etc., are inhibited by LPC. Triton X 100 which, by comparison, was used as a detergent effective on the guinea-pig small intestine, depressed the vasopressor effect of NE, PGF₂ and EPP.LPC, at low concentrations (40 mol/l), potentiated (15% max.) ADP-induced platelet aggregation (PA) in rat PRP but, at high concentrations, inhibited PA (IC₅₀=390 mol/l). 2-Hexadecylglycerophosphocholine and its short-chain 1-alkyl ethers, which are structurally related to platelet-activating factor, as well as some long-chain alkanol phosphocholine esters, were somewhat more active than LPC. Dipalmitoyllecithin (4–700 mol/l) was without any effect.     

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.     

Decreased serum levels of TGF-β in patients with acutePlasmodium falciparum malaria

Apart from cellular immunity and immunopathology, various cytokines have been implicated in malaria-associated immunosuppression. In this study, serum levels of transforming growth factor- (TGF-) were determined with an enzyme-linked immunosorbent assay in 37 patients with acutePlasmodium falciparum malaria prior to, during, and after therapy and in 17 healthy controls in Bangkok, Thailand. Patients were treated with artesunate and mefloquine. TGF- serum levels were found decreased prior to treatment (14±11 pg/ml versus 63±15 pg/ml in healthy controls;P<0.05). The serum concentrations of TGF- increased after initiation of treatment and were within normal range on day 21. Serum levels of both tumor necrosis factor-ga (TNF-) and soluble TNF-receptor 55 kDa were inversely correlated to serum levels of TGF- (r= –0.667 andr=}-0.592, n=37; respectively,P < 0.05 for both). No correlation between parasitemia and serum levels of TGF- could be found. The results are compatible with a decreased production and release, an enhanced clearance or utilization, or tissue accumulation of TGF- in acuteP. falciparum malaria.     

Efficacy of the two-microelectrode voltage clamp technique in crayfish muscle

Crayfish muscle fibres of different dimensions were voltage clamped and white noise current was injected into the fibres at various distances from the voltage clamp current electrode. The clamp current was measured and power spectral densities were calculated. This method revealed the efficacy of the voltage clamp in these fibres. In large fibres (l=1.8–2.0 mm; =100–180m) a space clamp was achieved only for a band width f=40Hz. At a distance of 100m from the clamp electrodes f was 250–500Hz. In fibres of medium size (l=1.0–1.3mm; =60–120m) f was about 80Hz and about 800 Hz at a distance of 100m. In experiments with very small muscle fibres (l=400–600m; =30–50m) f was more than 500Hz. The improvement of the space clamp for the smaller muscle fibres resulted mainly from the reduced total membrane capacity,c _m, of these fibres. The limitations of the space clamp could be derived from the impedance properties of the fibres. The band width of the space clamp correlated with the band width for which the square of the absolute impedance, |Z _p|², of the muscle fibre could be described by a simple RC-model. This correlation was demonstrated in a model circuit.Power density spectra of membrane current fluctuations were measured also. To optimize the resolution of these measurements the contribution of instrumental noise was minimized. The effects of instrumental noise are discussed.This investigation was supported by the Deutsche Forschungs-gemeinschaft     

Effects of GTPγS and fMet-Leu-Phe on rat PMN phospholipase C (PLC) activity

The PLC activities associated with the cytosol and plasma membrane subcellular fractions of rat PMN were tested for sensitivity to stimulation by GTPS and/or fMet-Leu-Phe. PMN plasma membrane PLC was stimulated 10–20% by 50 M GTPS (p<0.02). fMet-Leu-Phe alone had no effect on the plasma membrane PLC activity, but addition of fMet-Leu-Phe and GTPS together stimulated PLC activity by 20–30% (p<0.05). Neither GTPS nor fMet-Leu-Phe had any significant effect on the cytosolic PLC activity. These data demonstrate the presence of a PLC activity in rat PMN plasma membranes that is sensitive to stimulation by GTPS and fMet-Leu-Phe.     

Ca-dependent K channels in smooth muscle cells permeabilized by β-escin recorded using the cell-attached patch-clamp technique

Using the cell-attached patch-clamp technique, the activity of single, Ca-dependent K channels was recorded in single smooth muscle cells permeabilized by -escin. The conductance and the relationship between the open probability of the channels and pCa recorded in permeabilized cells were very similar to those obtained in excised inside-out patches. At pCa 7, application of 30 M acetylcholine (ACh) or 0.1 M substance P (SP) together with 1 mM guanosine 5-trisphosphate to permeabilized cells elicited transient bursts of channel openings similar to those which occur in intact cells. Transient activation was also observed when 2–30 M inositol trisphosphate (IP₃) was applied to permeabilized cells. This single channel activity was inhibited by pretreatment with low-molecular-weight heparin at 50–100 g/ml. Channel activity at pCa 7.0 was greatly enhanced by 200 M cyclic adenosine monophosphate. These results provide direct evidence that single Ca-dependent K channel activity is regulated by the transmitters ACh and SP, as well as a second messenger, IP₃, via the release of intracellular Ca from intracellular sites which are blocked by heparin. This novel approach is valuable in elucidating second messenger mechanisms involved in the regulation of single channel activity by transmitters and autacoids, since permeabilization by -escin preserves the entire system of receptor-operated signal transduction and allows intracellular application of second messengers at fixed concentrations.     

Scanning electron microscopy of merogonous stages ofEimeria falciformis var.pragensis inMus musculus

Asexual stages ofEimeria falciformis var.pragensis in Swiss-Webster mice were studied by scanning electron microscopy. Sporozoites were present in the cecum and colon 2 h post-inoculation (PI) and measured 11.3×2.1 m (9–13.9×2–2.2 m). Sporozoites penetrated epithelial cells with an extended anterior end and were constricted at the site of entry. Asexual generations were found in the cecum and colon epithelial cells. In meronts found at days 3–6 PI, merozoites matured synchronously, were oriented in the same direction, and were arranged in a helical pattern. Such meronts measured 11.3×6.4 m (8–13.7×5–7.4 m) and contained 8–12 meroxoites, which measured 11.9×1.5 m (7.4–15.7×1.3–1.8 m). Meronts which were present at day 7 PI measured 9.5×8.2 m (9–10.5×7–9.5 m) and contained 20–50 small merozoites which budded asynchronously from a central residuum. At days 3–7 PI, parasitized epithelial cells had shorter and fewer or no microvilli. The lumenal plasmalemma of the host cell was often disrupted or absent in cells containing mature meronts and escaping merozoites. At day 6 PI, phagocytic cells appeared on the epithelial surface, some of which were in contact with merozoites. Small foci of exposed basal lamina were present at day 7 PI in areas where cells had sloughed from the epithelium.     

Isolation and characterization of Chinese hamster ovary cell mutants resistant to the amino acid analogβ-aspartyl hydroxamate

Chinese hamster ovary cell lines which are resistant to an amino acid analog, -aspartyl hydroxamate, have been isolated and characterized. Mutants resistant to 100–150 M -aspartyl hydroxamate arose from ethyl methane sulfonate-treated parental lines at frequencies of 3.4×10^–6 to 1.3 ×10^–7. The mutants fell into at least two genetic classes: 18% of the mutants behaved codominantly in hybrids, the others recessively. Complementation studies indicated that all the recessive mutants belonged to the same class. Mutants selected after one step of mutagenesis overproduce the enzyme asparagine synthetase constitutively with four- to sixfold increases in specific activities over the basal levels of the parental lines. -Aspartyl hydroxamate-resistant cell lines with up to 20-fold elevations in asparagine synthetase activity have been isolated after two steps of mutagenesis. In addition, highly resistant lines have been selected by long-term growth of a dominant mutant in increasing concentrations of the drug. Resistance in the latter appears to be due not only to overproduction of asparagine synthetase but also to an alteration in the affinity of the enzyme for -aspartyl hydroxamate.     

Differential expression of β1, β3 and β4 integrins in sarcomas of the small,round, blue cell category

Integrins are a large and complex family of membrane spanning heterodimeric cell surface glycoproteins mediating cell/cell and cell/matrix interactions. Small, round, blue cell sarcomas (SRBCS) are a group of poorly differentiated tumours of various and in part uncertain histogenesis displaying similar cytomorphology. Among them are rhabdomyosarcomas (RMS), ganglioneuroblastomas [(G)NB], primitive peripheral neuroectodermal tumours (pPNET) and Ewing's sarcomas (ES). Thirty-two SRBCS were studied immunohistochemically for the distribution of 1, 3 and 4 integrins in situ. We found complex and to some extent differential patterns of 1, 3 and 4 integrin subunit expression in different types of SRBCS: all of the sarcomas studied were consistently 1⁺, 4^–, 2^–. Four of nine RMS were completely negative for all other integrin subunits studied while one RMS was 5⁺ throughout and three RMS were focally 5⁺. Three RMS expressed the 6 and v chains. In contrast to RMS, pPNET and ES, all of which were 1^–, 3^–, (G)NB were 3⁺ and frequently co-expressed 1. The eight pPNET and seven ES studied showed a similarily restricted integrin profile that was limited to the expression of 1 and 5 in nearly all cases. In summary, RMS were 1⁺, 1^–, 3^– and heterogeneously expressed 5 and 6. (G)NB were generally 1⁺, 1⁺, 3⁺, 5^–, 6^–. pPNET and ES were 1⁺, 1^–, 3^–, 5⁺, 6^–. The data illustrate a complex expression pattern of various integrins in SRBCS, a differential expression pattern of some of the integrin subunits among different types of SRBCS and almost identical integrin profiles in pPNET and ES.This paper is dedicated to Prof. Dr. Dres. h.c. Wilhelm Doerr on the occasion of his 80^th birthday     

Expression of surface membrane IgG on pokeweed mitogen-reactive anti-tetanus toxoid antibody-producing cells

Anti-tetanus toxoid antibody-producing cells, differentially expressing surface membrane IgM, were analyzed for the additional expression of surface membrane IgG. ⁺ and ^– cells were rosetted with anti--ox red blood cells and separated by density centrifugation into fractions enriched or depleted or ⁺ cells. These B-cell subsets were assayed for the production of IgM and IgG anti-tetanus toxoid antibody and total IgM and IgG. The results indicated that the majority of anti-tetanus toxoid antibody synthesis in the ^– fraction was by ⁺ cells. In the ⁺ fraction, however, both IgM and IgG anti-tetanus toxoid antibody production was detected in the ^+– and ⁺⁺ fraction. The inclusion of isotype-specific antisera during the first 2 days of culture further established that was expressed on the surface of the majority of the precursors for IgG anti-tetanus antibody productionin vitro. Studies performed to determine the culture requirements of ^– and ⁺ cells revealed that production of IgG anti-tetanus toxoid antibody by both cell subsets was dependent on T cells and pokeweed mitogen. However, some ^– cells could produce IgG in the presence of T cells alone.     

Quantal stores of excitatory transmitter in nerve-muscle synapses of crayfish evaluated from high-frequency asynchronous quantal release induced by veratridine or high concentrations of potassium

At single voltage-clamped opener muscle fibres of crayfish claw, 10–100 mol/l veratridine increased within a few seconds the rate of asynchronous quantal release, ñ, of excitatory transmitter from ñ<1 quantum/s to ñ10,000 quanta/s. Thereafter ñ declined exponentially either with a single, (2)50 s, or with two time constants (1)19 s, (2)50 s. In total (t), about 0.3 million quanta were released by veratridine in a single short fibre of about 1 mm length. These values were estimated by means of the noise analysis technique and they agreed with equivalent parameters of release when 100 mmol/l K⁺ were used as release stimulus. Strong quantal release could be elicited only once in a single muscle by veratridine. Furthermore, the effect of veratridine on quantal release could be completely prevented by pretreatment with tetrodotoxin. In another nerve-muscle preparation of crayfish, the abdominal superficial extensor muscle, up to 3 million excitatory quanta could be released by veratridine in a single fibre. In the latter muscle veratridine-induced asynchronous quantal release was strongly dependent on the extracellular concentration of Ca²⁺ whereas in the claw opener dependence of quantal release on extracellular Ca²⁺ was negligible.This investigation was supported by the Deutsche Forschungsgemeinschaft, SFB 220     

Intrinsic cholinergic excitation in the rat neostriatum: Nicotinic and muscarinic receptors

Summary An in vitro slice technique was employed to study the receptors involved in intrinsic cholinergic excitation in the rat neostriatum. The locally evoked synaptic potentials were suppressed by antinicotinic agents, mecamylamine (10 M), d-tubocurarine (3 M) or hexamethonium (100 M), but not by the antimuscarinic agent atropine (100 M). If the slices were exposed to an acetylcholinesterase (AChE)-inhibitor (paraoxon 1–20 M, physostigmine 0.1–0.5 M), the synaptic potentials were potentiated. The amplitude of the orthodromic population spike increased, and it was further facilitated when the stimulus frequencies were raised from 1–3 Hz to 10–30 Hz. The frequency facilitation following exposure to an AChE-inhibitor was blocked by atropine (1–100 M). Intracellular recording indicated that a slow depolarizing potential caused the frequency potentiation of the orthodromic discharges. Apparently rat neostriatum is similar to cholinergic systems in sympathetic ganglia and spinal Renshaw cells, in that nicotinic receptors mediate fast excitation and muscarinic receptors mediate slow excitation.     

Hb Tübingen. Eine neueβ-Kettenvariante (β Tp 10–12) mit erhöhter Spontanoxydation

Zusammenfassung Bericht über eine neue-Kettenvariante in einer in Süddeutschland ansässigen Familie. Von insgesamt sieben eingehend untersuchten Familienmitgliedern konnte die Anomalie in der heterozygoten Form bei zwei Angehörigen (Mutter und Sohn) nachgewiesen werden. Der Erbgang ist autosomal dominant. Das anomale Hämoglobin verursacht bei den Teilträgern infolge erhöhter Spontanoxydation eine Methämoglobinämie mit sichtbarer Cyanose sowie eine leichte kompensierte hämolytische Anämie ohne Innenkörperbildung. Der anomale Blutfarbstoff ist hitzelabil. Ein Enzymdefekt der Erythrocyten einschließlich Diaphorasemangel konnte ausgeschlossen werden. Globinanalysen ergaben einem im core ( Tp 10–12, Aminosäurenposition 83 bis 120) der-Kette gelegenen Defekt. Die Anomalie, die mit keiner der bisher bekannten Strukturvarianten übereinstimmt, wurde nach dem Ort des Auffindens als Hb Tübingen bezeichnet.

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Hb Tübingen. A new-chain variant ( Tp 10–12) showing increased spontaneous oxidation

Summary Hemoglobin Tübingen, a new-chain variant ( Tp 10–12) was discovered in two members out of seven of a German family. The abnormal hemoglobin is characterized by increased spontaneous oxidation, heat instability, normal methemoglobin spectrum, and the absence of spontaneous Heinz body formation. The carriers suffer from a mild and compensated hemolytic anemia; the main clinical symptom is a mild cyanosis. The amount of abnormal hemoglobin was approximately 40 per cent when DEAE-chromatography was performed with methemoglobin hemolysates; no separation could be achieved using hemolysates in the cyanmethemoglobin form. It was not possible to separate the abnormal-chain from normal-chain by globin electrophoresis or by chromatography on CM 52. Globin analyses showed a defect located in the core of the-chain ( Tp 10–12, amino acid position 83–120). This anomaly which does not correspond to any of the structural variants known so far was named Hb Tübingen according to the city in which it was found.

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Herrn Prof. Dr. K. H. Schäfer, Hamburg, zum 60. Geburtstag.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.