Prolonged blood pressure reduction by orally active renin inhibitor RO 42-5892 in essential hypertension.

OBJECTIVE--To investigate the effects of a novel specific renin inhibitor, RO 42-5892, with high affinity for human renin (Ki = 0.5 x 10(-9) mol/l), on plasma renin activity and angiotensin II concentration and on 24 hour ambulatory blood pressure in essential hypertension. DESIGN--Exploratory study in which active treatment was preceded by placebo. SETTING--Inpatient unit of teaching hospital. PATIENTS--Nine men with uncomplicated essential hypertension who had a normal sodium intake. INTERVENTIONS--Two single intravenous doses of RO 42-5892 (100 and 1,000 micrograms/kg in 10 minutes) given to six patients and one single oral dose (600 mg) given to the three others as well as to three of the patients who also received the two intravenous doses. RESULTS--With both intravenous and oral doses renin activity fell in 10 minutes to undetectably low values, while angiotensin II concentration fell overall by 80-90% with intravenous dosing and by 30-40% after the oral dose. Angiotensin II concentration was back to baseline four hours after the low and six hours after the high intravenous dose and remained low for at least eight hours after the oral dose. Blood pressure fell rapidly both after low and high intravenous doses and after the oral dose and remained low for hours. With the high intravenous dose the daytime (0900-2230), night time (2300-0600), and next morning (0630-0830) systolic blood pressures were significantly (p less than 0.05) lowered by 12.5 (95% confidence interval 5.6 to 19.7), 12.2 (5.4 to 19.3), and 10.7 (3.2 to 18.5) mm Hg respectively, and daytime diastolic pressure was lowered by 9.3 (2.2 to 16.8) mmHg. With the oral dose daytime, night time, and next morning systolic blood pressures were lowered by 10.3 (5.5 to 15.4), 10.5 (4.2 to 17.2), and 9.7 (4.0 to 15.6) mm Hg, and daytime and night time diastolic pressures were lowered by 5.8 (0.9 to 11.0) and 6.0 (0.3-12) mm Hg respectively. CONCLUSIONS--The effect of the inhibitor on blood pressure was maintained over a longer period than its effect on angiotensin II. RO 42-5892 is orally active and has a prolonged antihypertensive effect in patients who did not have sodium depletion. This prolonged effect seems to be independent, at least in part, of the suppression of circulating angiotensin II.     

Crystallization of trimeric recombinant human tumor necrosis factor (cachectin)

Crystals of tumor necrosis factor (TNF) have been obtained in two forms. Rhombohedral crystals grow in 1.8 to 2.0 M ammonium sulfite, pH 7.8 at 21 degrees C, and tetragonal crystals grow in 2.6 M magnesium sulfate, pH 5.5 at 25 degrees C. Analysis of TNF by isoelectric focusing under native and denaturing conditions indicates that TNF molecules exist as trimers in solution. The rhombohedral cachectin crystals belong to space group R3 and have unit cell constants a = b = c = 47.65 A and alpha = beta = gamma = 88.1 degrees. Density determinations and the space group indicate that the unit cell contains one 51,000-dalton trimer. These crystals are stable in the x-ray beam and diffract to at least 1.85 A but are apparently twinned by merohedry. The tetragonal crystals are space group P4(3)2(1)2 or its enantiomorph P4(1)2(1)2 and have unit cell constants a = b = 95.08, c = 117.49. The asymmetric unit contains one trimer; the crystals are stable in the x-ray beam and diffract to beyond 3 A.     

Release of L-alanine by tumor cells

Culture supernatants from the weakly immunogenic T cell lymphoma L5178Y ESb were found to contain substantial amounts of alanine and lactate at a ratio of about 1:10. Supernatants from cells of the highly immunogenic mutant line ESb-D also contained lactate but only minute amounts of alanine. Moreover, ESb cells converted 14C-labeled glucose or pyruvate into labeled alanine and lactate at a ratio of about 1:10, whereas ESb-D cells yielded only labeled lactate and no detectable alanine. The injection of L-alanine in combination with L-lactate into mice strongly suppressed the capacity of their spleen cells to generate cytotoxic responses. The injection of L-alanine also suppressed the immunogenicity of ESb-D cells, as demonstrated by the generation of cytotoxic activity in vivo and by the in vivo immunization (priming) for secondary cytotoxic responses against ESb-D cells in vitro. Taken together, these experiments suggest the possibility i) that the ESb cells prevent the induction of cytotoxic responses by releasing immunosuppressive alanine, and ii) that the immunogenic mutant ESb-D may have gained immunogenicity by losing this immunosuppressive property.     

Mapping of resistance to the potato cyst nematode Globodera rostochiensis from the wild potato species Solanum vernei

A population of diploid potato (Solanum tuberosum) was used for the genetic analysis and mapping of a locus for resistance to the potato cyst nematode Globodera rostochiensis, introgressed from the wild potato species Solanum vernei. Resistance tests of 108 genotypes of a F₁ population revealed the presence of a single locus with a dominant allele for resistance to G. rostochiensis pathotype Ro1. This locus, designated GroV1, was located on chromosome 5 with RFLP markers. Fine-mapping was performed with RAPD and SCAR markers. The GroV1 locus was found in the same region of the potato genome as the S. tuberosum ssp. andigena H1 nematode resistance locus. Both resistance loci could not excluded to be allelic. The identification of markers flanking the GroV1 locus offers a valuable strategy for marker-assisted selection for introgression of this nematode resistance.Abbreviations BSA bulked segregant analysis - RAPD random-amplified polymorphic DNA - RFLP restriction fragment length polymorphism - SCAR sequence-characterized amplified region     

A genetic map of potato (Solanum tuberosum) integrating molecular markers,including transposons,and classical markers

A genetic map of potato (Solanum tuberosum L.) integrating molecular markers with morphological and isozyme markers was constructed using a backcross population of 67 diploid potato plants. A general method for map construction is described that differs from previous methods employed in potato and other outbreeding plants. First, separate maps for the female and male parents were constructed. The female map contained 132 markers, whereas the male map contained 138 markers. Second, on the basis of the markers in common the two integrated parental maps were combined into one with the computer programme JoinMap. This combined map consisted of 175 molecular markers, 10 morphological markers and 8 isozyme markers. Ninety-two of the molecular markers were derived from DNA sequences flanking either T-DNA inserts in potato or reintegrated maize transposable elements originating from these T-DNA constructs. Clusters of distorted segregation were found on chromosomes 1,2,8 and 11 for the male parent and chromosome 5 for both parents. The total length of the combined map is 1120 cM.     

Antimicrobial peptides in the stomach of Xenopus laevis.

Antimicrobial peptides are widely distributed in nature and appear to play a role in the host defense of plants and animals. In this study we report the existence of antimicrobial peptides in the stomach of the vertebrate Xenopus laevis, an animal previously shown to store high concentrations of antimicrobial peptides in its skin. Antimicrobial activity was detected in extracts of X. laevis stomach tissue and nine antimicrobial peptides were then purified. A novel 24-amino acid peptide, designated PGQ, was isolated from these extracts, and has the following amino acid sequence: GVLSNVIGYLKKLGTGALNAVLKQ. PGQ is relatively basic and has the potential to form an amphipathic alpha-helix. The other peptides isolated are members of the magainin family of antimicrobial peptides, and include magainins I and II, PGLa, xenopsin precursor fragment, and four caerulein precursor fragments. None of these peptides had been previously identified in tissues other than the skin. The purification of the peptides from stomach extracts and subsequent protein sequence analysis reveals that the peptides have undergone the same processing as their dermal counterparts, and that they are stored in their processed forms. Northern blot analysis indicates that the magainin family of peptides are synthesized in the stomach, and immunohistochemical studies demonstrate that magainin is stored in a novel granular multinucleated cell in the gastric mucosa of Xenopus. This study demonstrates that the magainin family of antimicrobial peptides is found in the gastrointestinal system of X. laevis and offers an opportunity to further define the physiological role of these defense peptides.     

Pnoepyga immaculata n. sp., eine neue bodenbewohnende Timalie aus dem Nepal-Himalaya

Zusammenfassung Pnoepyga immaculata n. sp. lebt im Areal der beiden anderenPnoepyga-Artenalbiventer (Hodgson 1837) undpusilla Hodgson 1845 und wurde durch ihren markant abweichenden Territorialgesang entdeckt. Sie besitzt eine kurze silberhelle Strophe aus kurzen, in gleichmäßigen Abständen gereihten Pfiff-Elementen, die in der Frequenz leicht abfällt. Neben der Stimme liegen die wesentlichen diagnostischen Merkmale nach jetzigem Kenntnisstand gegenüber sympatrischenalbiventer in wenig größerem Schnabel, der Fleckenlosigkeit von Oberseite, Kopf und Flügeldecken, gegenüberpusilla in den größeren Körperabmessungen, gegenüber sympatrischenalbiventer undpusilla im schwachen grünlichen Schimmer der Oberseite und in der länglichen Schuppung der Unterseite. Es bestehen keine Proportionsunterschiede zwischenimmaculata undalbiventer/pusilla einerseits wie auch nicht zwischen diesen beiden andererseits. Bis jetzt wurden 4 Belegstücke (SammlungenKoelz, Diesselhorst, Martens) und eine durch Bandaufnahme gesicherte Beobachtung zwischen Dhaulagiri in Mittelwest-Nepal bis nahe der Ost-Grenze Nepals bekannt.P. immaculata lebt zur Brutzeit in enger Nachbarschaft mitP. albiventer undP. pusilla, ist jedoch vonalbiventer vertikal getrennt bei nachgewiesenem Kontakt, vonpusilla durch Bevorzugung trockeneren Waldunterwuchses abseits von Bächen. Schon jetzt mußimmaculata als gefährdete Art eingestuft werden, da Waldvegetation in der zur Brutzeit bevorzugten Höhenstufe zwischen 2100 und 3100 m in Nepal weitgehend vernichtet worden ist.

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Pnoepyga immaculata n. sp., a new ground living wren-babbler from the Nepal Himalayas (Timaliidae)

Summary P. immaculata n. sp. was discovered by its voice which strongly differs from the two other sympatric species,albiventer (Hodgson 1837) andpusilla Hodgson 1845. As a territorial song, it displays a silvery strophe arranged of short, more or less regularly spaced whistling notes (fig. 7a–d), slightly descending in pitch. The strophe is about 2 s long. Besides the distinct voice, the essential diagnostic characters in comparison toP. albiventer are the slightly larger bill, plain head, upper side and wing coverts (fig. 1a–d, 2, 4a–d), in comparison topusilla the larger body size (fig. 1i–m), in comparison to sympatricalbiventer andpusilla the slightly olive tinge of the upper side (against warm dark brown in both others, fig. 1e–m), and the more longish scaly feathers of the lower side (against stout scaly appearance in both others, fig. 1 a–d). These characters hold true at least in Central and Eastern Himalayan populations of the respective species. There do not exist proportional differences in the relations of any part of the body betweenimmaculata andpusilla and not between the latter ones as well. Heretofore, 4 museum specimens are known (Koelz Coll.,Diesselhorst Coll.,Martens Coll.) from West-central (Thakkhola, type locality) and East Nepal (Ting Sang La), 3 of them from the presumed breeding grounds (2100–3100 m), one from the winter quarters in the Terai lowlands. A field observation close to the Darjeeling border in far East Nepal is verified by a tape-recording.P. immaculata embarasses the biologist and systematist because it lives in close neighbourhood of the similar speciesalbiventer andpusilla. It must have been overlooked by its apparent scarcity and overall similarity to both other species of the genus. Slight ecological differences rather correspond to the vertical distribution ofalbiventer, which populates a higher forest belt. But both have already been found on territory at close range.P. pusilla inhabits different microbiotopes, especially the close proximity of running water in the same altitudinal belt asimmaculata. When discovered,P. immaculata is to be classified already as an endangered species. Forest vegetation of its preferred altitude is already greatly reduced in the Central and Eastern Nepal Himalayas. To get a more detailed information on its general distribution and biology in order to quickly arrange protected areas and thus to guarantee the survival ofP. immaculata, should be the urgent next step.

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Results of the Himalaya Expeditions ofJ. Martens, No. 163. — For No. 162 see: Stuttgarter Beitr. Naturkde. (A) 453: 1–46, 1990.     

Stable transformation of tomato cell cultures after bombardment with plasmid and YAC DNA

Summary Stable transformants were obtained after microprojectile particle bombardment of tomato cell suspensions (Lycopersicon esculentum cv VFNT Cherry and L. pennellii). The suspensions were bombarded with tungsten particles coated with either plasmid (6.3 kb) or yeast artificial chromosome (YAC) (80 kb) DNA containing the ß-glucuronidase (GUS) and neomycin phosphotransferase II (nptII) genes. The YAC DNA contained an insert of approximately 50 kb of DNA from VFNT Cherry. L. pennellii suspensions were more amenable to transformation than VFNT Cherry; more kanamycin-resistant calli were recovered from L. pennelli after bombardment with plasmid DNA, and only L. pennellii cells produced transformants after bombardment with YAC DNA. DNA gel blot analysis confirmed the presence of the nptll and GUS genes. This analysis also confirmed the integration of YAC DNA into the genome of the kanamycin-resistant calli and suggested that the level of intactness of the integrated YAC DNA was fairly high in four of the five transformants examined. Microprojectile bombardment of regenerable cultures with YACs may ultimately aid in map-based cloning of agriculturally-important genes.Abbreviations YAC yeast artificial chromosome - MS Murashige and Skoog - 2,4-D 2,4-dichlorophenoxy-acetic acid - IAA indole-3-acetic acid - GUS ß-glucuronidase - nptII neomycin phosphotransferase II     

Assessment of the potential bactericide effect of self‐cleaning floors: a proposed protocol

The antibacterial properties of self‐cleaning coatings are based on bactericide nanoparticles (NPs). Ecotoxicity of these NPs have been assessed mostly in suspension, using standard bioassays. Here a protocol is proposed to test actual coating samples, using the Vibrio fischeri bioluminescence inhibition bioassay. The protocol was designed to test bactericide properties of specially coated PVC floors being used in hospital environments under quasinatural conditions, such as prolonged exposure or room temperature. To take into consideration that the light output of the bacteria under prolonged exposure naturally changes, a correction factor is proposed.