Immunoelectron microscopic studies on the localization of peptidoglycan peptide subunit pentapeptides in bacterial cell walls

The peptide subunit pentapeptide H-L-Ala-D-Glu(L-Lys-D-Ala-D-Ala-OH)-NH₂ of peptidoglycan was localized in the cell walls of several Gram-positive bacteria employing the indirect immunoferritin technique. Specific antibodies to the D-alanyl-D-alanine moiety of non-crosslinked peptide subunit pentapeptide were raised in rabbits by immunization with synthetic immunogen albumin-(CH₂CO-Gly-L-Ala-L-Ala-D-Ala-D-Ala-OH)₃₉. Specificity of these antibodies for the peptide subunit pentapeptide and not for the peptide subunit tetrapeptide was corroborated in a Farr-type radio-active hapten binding assay. Specificity of labelling with ferritin was established by immunoelectron microscopic controls, and by the excellent correlation between specific labelling of cells with ferritin and the particular peptidoglycan primary structure of bacterial strains investigated. Cells of Lactobacillus gasseri, Streptococcus pyogenes and Staphylococcus aureus revealing non-crosslinked peptide subunit pentapeptides in their peptidoglycans could specifically be labelled. Lactobacillus acidophilus and Bacillus subtilis, on the contrary, missing such pentapeptides, failed in labelling.The implication of this method to possibly localize the points of attack of penicillin or cycloserine is discussed.Abbreviations used meso-A₂pm meso-diaminopimelic acid - DSM Deutsche Sammlung für Mikroorganismen, Göttingen, FRG This paper is dedicated to Professor Gerhart Drews on the occasion of his 60th birthday     

In vitro susceptibility ofPityrosporum ovale (Malassezia furfur) to human androgenic steroids

Cells ofPityrosporum ovale that colonize human pilosebaceous units are constantly exposed to cutaneous androgenic steroids. The aim of our study was to find out whetherP. ovale is susceptible to these hormones. Three strains ofP. ovale were grown in vitro in the presence of various concentrations oftestosterone, dehydroepiandrosterone, androstenedione, androstanedione, 5--dihydrotestosterone andprogesterone (10, 100, and 1000 µg/ml; agar dilution assays). In addition, three strains ofCandida albicans were also exposed to equal concentrations of the same androgens. As a result, allP. ovale strains were suppressed by 1000 µg/mlandrostenedione, which was the strongest inhibitor. The other androgenic steroids also significantly reducedP. ovale growth at different concentrations, depending on the hormone used and the strain tested.Progesterone was inhibitory at the highest concentration for oneP. ovale strain only.Candida albicans was not affected by any of the androgens. These findings demonstrate an in vitro susceptibility ofP. ovale to high concentrations of human androgenic steroids. A relevance of this interaction for the in vivo fungus-host relation is not apparent.     

Feinstruktur der Tarsal-Drüse vonSiro duricovius (Joseph) (Opiliones,Sironidae)

Zusammenfassung Alle Arten der Sironidae tragen als sekundäres männliches Geschlechtsmerkmal ein epidermales Drüsenorgan im letzten Tarsalglied des 4. Laufbeines. Das Sekret tritt dort dorsal, exponiert auf einer kanülenartigen Apophyse, dem Adenostyl, nach außen.BeiSiro duricorius (Joseph, 1868) besteht das komplexe Drüsenorgan aus zahlreichen funktionellen Einheiten, deren Sekret einem gemeinsamen Kanal zugeführt wird; er öffnet sich auf dem Adenostyl. Jede funktionelle Drüseneinheit umfaßt 5 Zellen: 3 Drüsenzellen (DZ, Dreiergruppe), 1 Hüllzelle (HZ), 1 Kanalzelle (KZ). Die Sekrete werden in ein gemeinsames Reservoir abgegeben.HZ verknüpft manschettenartig die DZ mit der KZ. Eine Randeinfaltung am Apex der DZ gewährleistet innige Verfalzung von diesen mit der HZ.DZ und HZ sind mit gut entwickeltem Mikrovillisaum ausgestattet; beide Zelltypen sezernieren.Der ableitende Kanal der GZ besteht aus 2 Abschnitten: ein distaler schwammig-fibrillärer und ein proximaler massiver; beide Teile werden von nur einer Zelle (KZ) begleitet. Der basale Teil mündet in den Hauptkanal, der zum Adenostyl zieht.Neben diesem komplexen Drüsenorgan liegt ein ähnlich gebautes, aber kleineres ventral im Tarsus; sein Sammelkanal zieht zum Kanal der Hauptdrüse.Isolierte funktionelle Einheiten sind in der Tarsenepidermis weit verstreut; sie leiten ihr Sekret unmittelbar auf die Tarsusoberfläche.Die DZ produzieren nach elektronenoptischer Evidenz ein Protein. Das Sekret der HZ läßt sich bisher nicht zuordnen; es ist von dem der DZ verschieden.

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Ultrastructure of the tarsal gland ofSiro duricorius (Joseph)

Summary All species of Sironidae possess as a secondary male sex character an epidermal gland organ in the last tarsal joint of the fourth leg. The glandular secretion emanates from a conical apophysis, the adenostyl, which is located dorsally on the joint.InSiro duricorius (Joseph, 1868) the complex organ consists of numerous functional units, the secretion of which flows into the common channel opening on the adenostyl.Every gland unit contains 5 cells: 3 gland cells (DZ), 1 enveloping cell (HZ), 1 duct cell (KZ). The enveloping cell connects the gland cells with the duct cell like a sleeve. A marginal fold of the gland cell apex provides tight connection of gland cells and enveloping cell.Gland cells and enveloping cell are equipped with microvilli, and both cell types secrete in a common reservoir.The duct of the duct cell is divided into two parts: 1) a distal fibrillaceous-spongy one and 2) a proximal massive one. Both are accompanied only by a single duct cell.Besides this complex gland organ a similar one is located in the ventral part of the tarsus. Its collecting channel opens into the channel of the main gland.Isolated functional gland units are scattered over the epidermis of the tarsus; their secretion flows immediately on the tarsal surface.The gland cells proper produce a protein, the fact is indicated by the electronmicrographs. The secretion of the enveloping cell is a different one, but at this time cannot be attached to a chemical group.

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Abkürzungen A Adenostyl - aSe austretendes Sekret - C Kutikulin - Cu Kutikula - dK distaler Kanal - DK Kern der Drüsenzelle - DZ Drüsenzelle - ER rauhes endoplasmatischesm, Retikulum - G Golgi-Apparat - HK Kern der Hüllzelle - HZ Hüllzelle - K Kralle des Tarsus - Ke Kern - KK Kern der Kanalzelle - KZ Kanalzelle - M Mikrovilli - Mi Mitochondrium - pK proximaler Kanal - R Randeinfaltung der DZ - Re extrazelluläres Reservoir - Ri Ribosomen - Se Sekret - SK zentraler Sammelkanal - T Mikrotubuli - Va Vakuole - Va₁-Va₆ Vakuolen mit Sekret unterschiedlichen Reifegrades     

Key Elements in a Framework for Land Use Impact Assessment Within LCA (11 pp)

Background, Aim and Scope Land use by agriculture, forestry, mining, house-building or industry leads to substantial impacts, particularly on biodiversity and on soil quality as a supplier of life support functions. Unfortunately there is no widely accepted assessment method so far for land use impacts. This paper presents an attempt, within the UNEP-SETAC Life Cycle Initiative, to provide a framework for the Life Cycle Impact Assessment (LCIA) of land use. Materials and Methods: This framework builds from previous documents, particularly the SETAC book on LCIA (Lindeijer et al. 2002), developing essential issues such as the reference for occupation impacts; the impact pathways to be included in the analysis; the units of measure in the impact mechanism (land use interventions to impacts); the ways to deal with impacts in the future; and bio-geographical differentiation. Results: The paper describes the selected impact pathways, linking the land use elementary flows (occupation; transformation) and parameters (intensity) registered in the inventory (LCI) to the midpoint impact indicators and to the relevant damage categories (natural environment and natural resources). An impact occurs when the land properties are modified (transformation) and also when the current man-made properties are maintained (occupation). Discussion: The size of impact is the difference between the effect on land quality from the studied case of land use and a suitable reference land use on the same area (dynamic reference situation). The impact depends not only on the type of land use (including coverage and intensity) but is also heavily influenced by the bio-geographical conditions of the area. The time lag between the land use intervention and the impact may be large; thus land use impacts should be calculated over a reasonable time period after the actual land use finishes, at least until a new steady state in land quality is reached. Conclusions: Guidance is provided on the definition of the dynamic reference situation and on methods and time frame to assess the impacts occurring after the actual land use. Including the occupation impacts acknowledges that humans are not the sole users of land. Recommendations and Perspectives: The main damages affected by land use that should be considered by any method to assess land use impacts in LCIA are: biodiversity (existence value); biotic production potential (including soil fertility and use value of biodiversity); ecological soil quality (including life support functions of soil other than biotic production potential). Bio-geographical differentiation is required for land use impacts, because the same intervention may have different consequences depending on the sensitivity and inherent land quality of the environment where it occurs. For the moment, an indication of how such task could be done and likely bio-geographical parameters to be considered are suggested. The recommendation of indicators for the suggested impact categories is a matter of future research.     

Preferential Colonization of Metastases by Oncolytic Vaccinia Virus Strain GLV-1h68 in a Human PC-3 Prostate Cancer Model in Nude Mice

Recently, we showed that the oncolytic vaccinia virus GLV-1h68 has a significant therapeutic potential in treating lymph node metastases of human PC-3 prostate carcinoma in tumor xenografts. In this study, underlying mechanisms of the virus-mediated metastases reduction were analyzed. Immunohistochemistry demonstrated that virus-treatment resulted in a drastically decrease of blood and lymph vessels, representing essential routes for PC-3 cell migration, in both tumors and metastases. Thus, GLV-1h68 drastically reduced essential routes for the metastatic spread of PC-3 cells. Furthermore, analysis of viral distribution in GLV-1h68-injected tumor-bearing mice by plaque assays, revealed significantly higher virus titers in metastases compared to solid tumors. To elucidate conditions potentially mediating the preferential viral colonization and eradication of metastases, microenvironmental components of uninfected tumors and metastases were compared by microscopic studies. These analyses revealed that PC-3 lymph node metastases showed increased vascular permeability, higher proliferation status of tumor cells as determined by BrdU- and Ki-67 assays and lesser necrosis of PC-3 cells than solid tumors. Moreover, an increased number of immune cells (MHCII⁺/CD68⁺ macrophages, MHCII⁺/CD19⁺ B lymphocytes) combined with an up-regulated expression of pro-inflammatory cytokines was observed in metastases in comparison to primary PC-3 tumors. We propose that these microenvironmental components mediated the metastatic tropism of GLV-1h68. Therefore, vaccinia virus-based oncolytic virotherapy might offer a novel treatment of metastatic prostate carcinomas in humans.     

Hybrid maize breeding with doubled haploids: I. One-stage versus two-stage selection for testcross performance

Optimum allocation of resources is of fundamental importance for the efficiency of breeding programs. The objectives of our study were to (1) determine the optimum allocation for the number of lines and test locations in hybrid maize breeding with doubled haploids (DHs) regarding two optimization criteria, the selection gain ΔG _k and the probability P _k of identifying superior genotypes, (2) compare both optimization criteria including their standard deviations (SDs), and (3) investigate the influence of production costs of DHs on the optimum allocation. For different budgets, number of finally selected lines, ratios of variance components, and production costs of DHs, the optimum allocation of test resources under one- and two-stage selection for testcross performance with a given tester was determined by using Monte Carlo simulations. In one-stage selection, lines are tested in field trials in a single year. In two-stage selection, optimum allocation of resources involves evaluation of (1) a large number of lines in a small number of test locations in the first year and (2) a small number of the selected superior lines in a large number of test locations in the second year, thereby maximizing both optimization criteria. Furthermore, to have a realistic chance of identifying a superior genotype, the probability P _k of identifying superior genotypes should be greater than 75%. For budgets between 200 and 5,000 field plot equivalents, P _k > 75% was reached only for genotypes belonging to the best 5% of the population. As the optimum allocation for P _k (5%) was similar to that for ΔG _k , the choice of the optimization criterion was not crucial. The production costs of DHs had only a minor effect on the optimum number of locations and on values of the optimization criteria. C. Friedrich H. Longin and H. Friedrich Utz contributed equally to this work.