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.     

Nachweis von 6 neuen Steroiden im Plasma von menschlichem Placentablut (Nabelsehnurblut)

Zusammenfassung 16-Hydroxyprogesteron, 17-Hydroxyprogesteron, ⁴-Pregnen-20-ol-3-on, ⁴-Pregnen-17, 20-diol-3-on, Adrenosteron und 11-Hydroxyandrostendion wurden in Extrakten von Plasma des menschlichen Placentablutes (Nabelschnurblut) nachgewiesen.     

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).     

Genetic characterization of an α-specific gene responsible for sexual agglutinability in Saccharomyces cerevisiae: mapping and gene dose effect

Summary A recessive ag1 mutation leads to specific defect in sexual agglutinability specifically in cells of the yeast Saccharomyces cerevisiae. The cryptopleurine resistance gene cry^R 1, closely linked to the mating type locus, was used to select / strains which emerged from / strains by mitotic nonreciprocal recombination, to genetically analyse ag1, since ag1 is expressed only in mating type. The ag1 gene was found to be linked to the centromere tightly, to met3 at 4.4 cM, and to ilv3 at 12 cM on chromosome X. Sexual agglutinability of cells was shown to be dependent on the dose of the AG1 gene, using / isogenic strains carrying AG1/AG1, AG1/ag1 or ag1/ag1. The sst2-1 mutation did not suppress the ag1 mutation. Based on these results, function of the AG1 gene is discussed.Abbreviations cM centimorgan - FDS first division segregation - NPD nonparental ditype - PD parental ditype - SDS second division segregation - TT tetratype     

Synergistic effects of tumour necrosis factor-α and interferon-γ on feline haemopoietic progenitors

Pathogenic mechanisms that underlie feline leukaemia virus subgroup-C (FeLV-C) induced erythroid aplasia are unknown. FeLV-C infection is associated with higher serum levels of interferon- (IFN-) and tumour necrosis factor- (TNF-), which may act synergistically to cause haemopoietic suppression. In the present studies, the synergistic effects of TNF- and IFN- on feline bone marrow progenitors in vitro were evaluated. Bone marrow mononuclear cells from specific-pathogen-free cats were exposed to TNF- (100 and 200 pg/ml) and IFN- (100 or 200 units/ml), alone or in combination, for 2 h before plating for clonal assays of colony forming units. Our results show that TNF- and IFN- in combination caused marked suppression of feline colony forming units-erythroid (CFU-E), burst forming units-erythroid (BFU-E), and colony forming units-fibroblasts (CFU-F), whereas colony forming units-granulocyte/macrophage (CFU-GM) were minimally affected. The same concentrations of TNF- and IFN- alone had minimal effects on CFU-E, BFU-E and CFU-F. These results suggest that TNF- and IFN- may play a significant role in regulating haemopoiesis in cats and may be involved in the pathogenesis of erythroid aplasia in cats infected with feline leukaemia virus.     

Effect of dehydroepiandrosterone on radioligand binding of [<Superscript>3</Superscript>H]-testosterone by androgen receptors in rat hypothalamus

Intramuscular injections of dehydroepiandrosterone in a dose of 0.7 mg/kg for 10 days significantly increased nuclear and cytoplasmic fractions of androgen receptors in the preoptic/ anterior hypothalamic area. Presumably, the effect of the neurosteroid is mediated by 5-reductase transformation of dehydroepiandrosterone into 5-dehydroepiandrosterone, which initiates the synthesis of androgen receptors.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 10, pp. 435–438, October, 2004     

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.     

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     

Sensitivity to dihydropyridines, ω-conotoxin and noradrenaline reveals multiple high-voltage-activated Ca2+ channels in rat insulinoma and human pancreatic β-cells

High-voltage-activated (HVA) Ba²⁺ currents of rat insulinoma (RINm5F) and human pancreatic -cells were tested for their sensitivity to dihydropyridines (DHPs), -conotoxin (-CgTx) and noradrenaline. In RINm5F cells, block of HVA currents by nimodipine, nitrendipine and nifedipine was voltage- and dose-dependent (apparent K _D<37 nM) and largely incomplete even at saturating doses of DHPs (mean 53%, at 10 M and 0 mV). Analysis of slow tail currents in Bay K 8644-treated cells indicated the existence of Bay K 8644-insensitive channels that turned on at slightly more positive voltages and deactivated more quickly than Bay K 8644-modified channels. DHP Ca²⁺ agonists and antagonists in human -cells had similar features to RINm5F cells except that DHP block was more pronounced (76%, at 10 M and 0 mV) and Bay K 8644 action was more effective, suggesting a higher density of L-type Ca²⁺ channels in these cells. In RINm5F cells, but not in human -cells, DHP-resistant currents were sensitive to -CgTx. The toxin depressed 10–20% of the DHP-resistant currents sparing a residual current (25–35%) with similar voltage-dependent characteristics and Ca²⁺/Ba²⁺ permeability. Noradrenaline (10 M) exhibited different actions on the various HVA current components: (1) it prolonged the activation kinetics of -CgTx-sensitive currents, (2) it depressed by about 20% the size of DHP-sensitive currents, and (3) it had little or no effects on the residual DHP- and -CgTx-resistant current although intracellularly applied guanosine 5-O-(3-thiotriphosphate) (GTP--S) prolonged its activation time course. The first action was clearly voltage-dependent and most evident in RINm5F cells that displayed neuronal-like processes. The second was observed more frequently, was voltage-independent and fully blocked by saturating doses of nifedipine (10 M). Both actions were prevented by intracellular perfusion with guanosine 5-O-(2-thiodiphosphate) (GDP--S). Our data suggest that beside a majority of L-type channels, RINm5F and human pancreatic -cells may express a variable fraction of DHP-insensitive channels that may be involved in the control of insulin secretion during -cell activity.     

Antikatatoxische Hormone

Zusammenfassung Nach einer kurzen Übersicht der Literatur über den Einfluß von Hormonen auf die Resistenz werden die grundlegenden Unterschiede zwischen syntoxisch und katatoxisch wirkenden Hormonen beschrieben. Die ersteren (z. B. die Glucocorticoide) greifen die schädigenden Stoffe selbst nicht an, unterdrücken jedoch die Reaktionen der Gewebe (z. B. Entzündung). Demgegenüber beeinflussen katatoxische Hormone die Gewebsreaktion nicht, sie beschleunigen jedoch den Abbau pathogener Stoffe. Gewöhnlich sind die Abbauprodukte weniger toxisch als der ursprüngliche Giftstoff, und in diesen Fällen üben katatoxische Hormone eine Schutzwirkung aus.Die neuesten, hier ausführlich beschriebenen Versuche zeigen jedoch, daß die Schutzwirkung katatoxischer Hormone, z. B. des Pregnenolon-16-carbonitrils (PCN) und des Spironolactons (SNL) sowohl gegen Digitoxin als auch gegen Indomethazin durch Triamcinolon und Thyroxin unterdrückt werden können. Oestradiol ist in dieser Beziehung nur gegen Digitoxin wirksam. Eine derartige echte antikatatoxische Wirkung hängt nicht schlechtweg von der Steroidstruktur der Wirkstoffe ab, da Desoxycorticosteron (DOC), Progesteron und Hydroxydion unter gleichen Versuchsbedingungen weder gegen Digitoxin noch gegen Indomethazin die Schutzwirkung von PCN oder SNL unterdrücken, während Thyroxin diesbezüglich hoch wirksam ist.Die im Text erwähnten Arbeiten wurden vom Medical Research Council of Canada (Block Term Grant MT-1829), dem Ministère des Affaires Sociales, Québec, der Quebec Heart Foundation und der Succession J. A. DeSève unterstützt. — Die folgenden Substanzen wurden in großzügiger Weise von den Firmen Upjohn (PCN), Searle (SNL), Squibb (Triamcinolon), Organon (DOC-Acetat), Roussel (Oestradiol, Progesteron) und Pfizer (Hydroxydion) zur Verfügung gestellt.     

Anticipatory postural changes induced by active unloading and comparison with passive unloading in man

Normal human subjects, sitting in a chair, were required to maintain stable elbow flexion against loads of 0.5 kg or 1.0 kg. Unloading was affected either passively by the experimenter, or actively with the subject's own contralateral arm. Elbow angle, force exerted by the load, and electromyographic activity (EMG) of biceps and triceps muscles of both arms were recorded and averaged. Passive unloading was followed by a reduction of biceps EMG activity, starting 50–80 ms after weight lift, and by an upward deflection of the forearm. With active unloading, however, a reduction of the biceps EMG activity slightly preceded the onset of unloading (0–30 ms). This reduction of the actively unloaded arm occurred at about the same time as the activity of the contralateral unloading arm. In this experiment, the unloaded forearm maintained an almost stable position. Thus, the anticipatory adjustment of elbow posture, observed when unloading was performed by the subject, appears to optimize limb stability during the mechanical perturbation.     

Lichtmikroskopische Untersuchungen über die Entwicklung vonTheileria annulata (Dschunkowsky und Luhs, 1904) inHyalomma anatolicum excavatum (Koch, 1844)

Summary Fully differentiated kinetes, average length 17.6m, appeared in the haemolymph of engorged nymphs usually 17 to 20 days after repletion. Kinetes were observed at first in the salivary glands on day 18 after repletion. The kinetes then transformed into fission bodies of about 10m in diameter, mainly in type III alveoli and less frequently in type II alveoli. The fission bodies grew up to a size of about 20m after several divisions of their nucleus. At this time the ticks moulted and no further development occurred until activation. Shortly before infestation the salivary glands began to proliferate, and rapid growth of the fission bodies was observed, especially in young ticks where development of infective particles (sporozoites) was concluded within two days. Development in feeding adult ticks apparently occurred in four major steps: (1) Division of primary fission bodies (sporonts) into numerous secondary fission bodies (primary sporoblasts), (2) division of secondary fission bodies into tertiary fission bodies (secondary sporoblasts), (3) production of particles (sporozoites) by tertiary fission bodies and release of particles into the saliva, and (4) degeneration of fission bodies and their host cell but further release of particles.The host cell was stimulated to giant growth, thus its diameter increased, on average, from 15 to 110 m. Heavy infections resulting from parasitaemias of >40% caused disease and mortality in the tick population. Development was much retarded by aging. In ticks starved for six months sporozoites did not develop before day five to seven of infestation. Sporozoites may not develop at all in six to nine month old female ticks during the infestation period. The significance of the described developmental stages ofT. annulata was discussed and a sexual generation postulated. The hypothetic development ofT. annulata in its tick vector was illustrated.

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Gefördert von der Deutschen Forschungsgemeinschaft (DFG)     

Ultrasonography gives directly but noninvasively elastic characteristic of human tendon in vivo

To obtain an insight into tendon elasticity during human movement, a real-time ultrasonography was applied to the contracting tibialis anterior muscle. The insertion point of fascicles onto the aponeurosis was clearly visualized, and its position relative to a fixed marker on the skin moved proximally (1) according to the increasing dorsiflexion force (F) with a fixed ankle joint. Notably, the 1 – F relationship in the tendon was found to be quadratic in nature (F = c1²; c=1.48 2.24, r=0.985 0.992, n=9) as has been reported in the isolated tendon, although the F – 1 curves were slightly underestimated in comparison with the stiffness constant estimated from tendon architecture. This underestimation might be caused by changes in the height of the foot arch with the application of force.     

Assessing skills for refusing cigarettes and smokeless tobacco

Hops and colleagues developed an audiotaped refusals skills test in which students respond to cigarette offers and their responses are scored for content. The present study employed a modified analogue skills test. Modifications included adding a separate subscale for smokeless tobacco, emphasizing repeated offers and group pressure, and rating the quality of responses (good, fair, poor). The test was evaluated in four seventh-grade classrooms (N=78). Half had participated in a refusals skills training program; the others were controls. Intervention subjects provided more good responses and fewer poor responses than controls. In a multiple regression, repeated and group offers were associated with the quality of response, while offerer's gender and type of tobacco variables were not associated. In a second regression, experimental condition was associated with quality of the responses, while gender, ethnicity, exposure to tobacco, use of tobacco, and attitudes toward the test were not associated.This work was supported by Grant R01-CA44921 from the National Cancer Institute to John P. Elder.     

Phosphorylation-regulated low-conductance Cl− channels in a human pancreatic duct cell line

A low-conductance Cl^– channel has been identified in the apical membrane of the human pancreatic duct cell Capan-1 using patch-clamp techniques. Cell-attached channels were activated by the vasoactive intestinal polypeptide (VIP, 0.1 mol/l), dibutyryl-adenosine 3,5-cyclic monophosphate (db-cAMP, 1 mmol/l), 8-bromo adenosine 3,5-cyclic monophosphate (8-BrcAMP, 1 mmol/l), 3-isobutyl-1-methyl-xanthine (IBMX, 100 mol/l) and forskolin (10 mol/l). No channel activity was observed in non-stimulated control cells. In both cell-attached and excised inside-out patches, the channel had a linear current/voltage relationship and a unitary conductance of 9 pS at 23°C and 12 pS at 37°C. Its opening probability was not voltage dependent although pronounced flickering was induced at negative potentials. Anionic substitution led to the selectivity sequence Cl^–>I^–>HCO₃ ^–>gluconate. In insideout excised patches, the channel activity declined spontaneously within a few minutes. Reactivation of silent excised channels was achieved by adding protein kinase A (PKA, in the presence of ATP, cAMP and Mg²⁺). Conversely, active channels were silenced in the presence of alkaline phosphatase. The PKA-activated Cl^– channel was 4,4-diisothiocyanatostilbene-2,2-disulphonic acid (DIDS, 100 mol/l) and 4-acetamido-4-isothiocyanatostilbene-2, 2-disulphonic acid (SITS, 100 mol/l) insensitive, but was blocked by diphenylamine-2-carboxylic acid (DPC, 100 mol/l). These results demonstrate that the apical low-conductance Cl^– channel in Capan-1 is regulated on-cell by VIP receptors via cAMP and off-cell by PKA and phosphatases. They provide evidence that this channel is closely related to the cystic fibrosis transmembrane conductance regulator (CFTR) Cl^– channel.     

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.     

Diffusionsfehler und Eigenverbrauch der Pt-elektrode beipO2-Messungen im steady state

Zusammenfassung Im steady state beeinflussen Diffusionsfehler und Eigenverbrauch der Pt-Elektrode als systematische Fehler O₂-Partialdruckmessungen. Sie sind abhängig von den geometrischen Eigenschaften der Elektrode, den Diffusionseigenschaften der Membran sowie den Diffusions- und Konvektionseigenschaften des Meßmediums. Das Diffusionsfeld vor der Pt-Oberfläche und das dadurch bestimmte stationäre Meßsignal werden für gasförmige und nicht gasförmige Medien mit und ohne Konvektion berechnet. Daraus resultieren quantitative Aussagen über die systematischen Fehler. Speziell für Messungen in durchbluteten Geweben (z. B. Hirnrinde und Myokard) wird der Einfluß des Eigenverbrauchs von Pt-Elektroden auf den intracapillärenpO₂-Abfall in Durchblutungsrichtung und das intercapillärepO₂-Feld am Meßort der Elektrode ermittelt. Diese Berechnungen erfolgten mit Hilfe eines Digitalmodells.

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Erklärung der Symbole A O₂-Verbrauch des Gewebes - Bunsenscher Löslichkeitskoeffizient des Mediums - _m Bunsenscher Löslichkeitskoeffizient der Membran - C ₁,C ₁,C ₂,C ₂,C ₃ Konstanten - D Diffusionskoeffizient des Mediums - DF, DF Diffusionsfehler bei einfacher und doppelter Membran - DGl Differentialgleichung - d Capillarabstand - d _h Dicke der hydrodynamischen Grenzschicht - d _m ,d _m Dicke der Membranen - , , , , , dimensionslose Parameter - exp Exponentialfunktion - F Faradaykonstante - grad Gradient - I _o stationäres Meßsignal in Medien ohne Konvektion - I stationäres Meßsignal in Gasen - I _o stationäres Meßsignal in Flüssigkeiten mit Konvektion - J _o nullte Bessel-Funktion - K Diffusionsleitfähigkeit des Mediums - KE, KE Konvektionseffekt bei einfacher und doppelter Membran - K _m ,K _m Diffusionsleitfähigkeit der Membranen - l Capillarlänge - l Capillarabschnitt - Viscosität des Mediums - p, pO₂,p(r), p(r,z) O₂-Partialdruck - p mittlerer Partialdruck - P _a O₂-Partialdruck am arteriellen Capillarende - p _c konstanter Partialdruck - P/r _o +d _m O₂-Partialdruck an der Grenze Membran/Medium - P _v O₂-Partialdruck am venösen Capillarende - P _g relativer O₂-Partialdruckabfall im Gewebe - P _v relativer O₂-Partialdruckabfall am venösen Capillarende - R Radius der ebenen kreisförmigen Elektrode - RB Randbedingung - RDF, RDF restlicher Diffusionsfehler einfacher und doppelter Membranen - r _o Radius der Elektrode mit halbkugelförmiger Pt-Oberfläche - r, z Zylinderkoordinaten - r _K Capillarradius - S Sättigungsabfall im Capillarblut ohne Elektrode - S Sättigungsabfall im Capillarblut mit Elektrode - u O₂-Konzentration - V Diffusionsgesamtfluß - V _K Diffusionsfluß aus einem Capillarabschnitt - v _r ,v _z Komponenten des Stromdichtevektors inr- bzw.z-Richtung (Zylinderkoordinaten) - mittlere Stromdichte - Stromdichtevektor des Flusses der O₂-Moleküle - v _c konstante Geschwindigkeit des bewegten Mediums - x, y, z Kartesische Koordinaten - Integrationsvariable - ² Laplace-Operator - partielle Ableitung nach der Zeit     

Selection of E. coli strains for stable transformation with recombinant plasmids containing full-length genome of clinical HIV-1 isolates

Strain 6007 obtained from the parent E. coli strain 5097 is a result of ptsH5 mutation, which allowed cells to grow without common components of the phosphoenolpyruvate-dependent phosphotransferase system. Segregants of strain 6007 retaining the Pol⁺ gene responsible for inability to grow at 37°C, but gaining rifampicin resistance (Rif^R) were used for cloning of cointegrate plasmids. Preintegration complexes of HIV-1 were cointergated with the pBR-322 plasmid and transformed strain 6018. Sequencing showed that the pPIC91 hybrid plasmid contains full-length genome of HIV-1 with shortened 5-terminal LTR and full-length copy of pBR322. Elimination of the pPIC91 plasmid from 6018 cells was followed by the appearance of auxotrophic insertion mutants. Sequencing of the insert region showed that chromosome DNA of the host cell includes integrated genomes of pBR-322 and HIV-1.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 11, pp. 550–554, November, 2004     

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.     

Immunochemical identification of an α2-macroglobubin specific for the human kidney

An ₂-macroglobubin specific for the kidney (RS ₂-macroglobubin) with a molecular weight of 2×10⁶, was identified by immunochemical analysis. The antigen was shown not to be immunochemically identical with ₂-H-globubin (ferritin), renal-pancreatic ₂-globubin, uromucoid, or serum ₂-macroglobubin. The content of RS ₂-macroglobubin in kidney tumor tissue was lower than in the normal kidney and none whatever was found in 16 of 23 tumors. At the level of sensitivity of the monospecific test system for RS ₂-macroglobubin it could not be found in healthy human blood or in the blood and urine of patients with nephrological diseases.Department of Urology and Operative Nephrology, N. I. Pirogov Second Moscow Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Lopatkin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 8, pp. 210–213, August, 1977.