Characterization of the state of differentiation of six newly established human non-small-cell lung cancer cell lines

Six new non-small-cell lung cancer (NSCLC) cell lines were established directly from human tissue or indirectly via nude mouse xenografts in serum-supplemented media with success rates of 8% and 13%, respectively. They comprised one adenocarcinoma (ADLC-5M2), two squamous cell carcinomas (EPLC-32M1, EPLC-65H), two large cell carcinomas (LCLC-97TM1, LCLC-103H), and one malignant biphasic mesothelioma (MSTO-211H). All cell lines grew adherent to culture vessels with population doubling times (PDT) of 16-40 h, formed colonies in soft agarose with efficiencies of 0.1%-5.1%, and all grew in athymic nude mice. Xenograft histologies appeared as follows: (a) undifferentiated carcinomas with feeble resemblance to the original tumors in the case of adenocarcinomas and squamous cell carcinomas; (b) large cell carcinoma with high resemblance to the original tumor; (c) an undifferentiated tumor with predominance of large epithelial cells and few fibrous cells in the case of mesothelioma. Human chorionic gonadotropin (HCG) was found by radioimmunoassay and high-affinity binding sites for epidermal growth factor (EGF) by radio-receptor assay in 4/4 cell lines. A very low activity of L-DOPA decarboxylase (DDC) was detectable only in the adenocarcinoma cell line. All cell lines overexpressed the c-myc protooncogene, and no gene rearrangement or amplification was observed. Chromosome analysis revealed modal chromosome numbers of 70-73 in ADLC-5M2, EPLC-32M1, EPLC-65H, and MSTO-211H. Cell lines derived from large cell carcinoma had modal values of 65 and 170 and a wider chromosome distribution than all other cell lines. A NSCLC specific chromosomal aberration has been undetectable until now. These cell lines may aid in elucidating the biology of NSCLC and its interrelationship to other lung tumors.     

Substance P analogues function as bombesin receptor antagonists and inhibit small cell lung cancer clonal growth

Human small cell lung cancer (SCLC) produces and secretes BN/GRP (bombesin/gastrin releasing peptide). Because BN stimulates the growth of SCLC cells and these cells have receptors for BN-like peptides, it is important to define agents which disrupt this self-promoting autocrine growth cycle. Here, substance P analogues were evaluated as BN receptor antagonists using SCLC cell lines. (D-Arg¹, D-Pro², D-Trp^7,9, Leu¹¹) substance P [(APTTL)SP] was one of the more potent analogues tested in inhibiting BN-like peptide receptor binding with an IC₅₀ value of 1 μM. Micromolar concentrations of (APTTL)SP antagonized BN receptor mediated elevation of cytosolic Ca²⁺ levels and decreased the colony formation in soft agarose. These data suggest that SP analogues function as SCLC BN receptor antagonists and may be useful in disrupting the autocrine growth function of BN-like peptides.     

Models of the geometry of the human ankle bone pulley: two series of geometric models for demonstrating the biomechanics of the ankle joint

2 series of models demonstrate the geometrical shape of the human trochlea tali. We have changed step by step the shape of the 2 flanking articular facets of the trochlea, the course of the edges of the trochlea, and the length of their radii, and so we have found a model responding to the biomechanic conditions of the trochlea tali. The convex surface of this model (corresponding to the superior articular surface, i.e. the facies superior trochleae tali) is a torse, the medial flanking facet (corresponding to the medial articular facet of the trochlea, i.e. the facies malleolaris medialis) is a flat cone, the lateral flanking facet (corresponding to the lateral articular facet of the trochlea, i.e. the facies malleolaris lateralis) is a screwed (helicoidal) face. The resulting model shows the 2 completely different phases of motion in the ankle joint: During dorsiflexion (motion setting out from the neutral position towards the final position of dorsiflexion), the internal malleolus leads the talus, whereas the external malleolus is pushed outwards by the screwed lateral articular facet of the trocheal. The trochlea is moved like a hinge. In the final position of dorsiflexion, the malleoli tightly embrace the 2 flanking facets of the trochlea, whilst an obvious cleft appears dorsally and medially between the superior articular surface of the trochlea and the tibial roof (i.e. the facies articularis inferior tibiae). During plantarflexion (motion setting out from the neutral position towards the final position of plantarflexion), the external malleolus leads the talus, whereas the medial articular facet of the trochlea withdraws from the internal malleolus. The trochlea is moved like a screw. In the final position of plantarflexion, the superior articular surface of the trochlea closely contacts the tibial roof, whilst an obvious cleft appears between the medial articular facet of the trochlea and the internal malleolus.     

Elicitation of volatile compounds in photomixotrophic cell culture of Petroselinum crispum

Photomixotrophic cells of Petroselinum crispum accumulated >500 mg chlorophyll per kg wet weight and grew well in a broad range of phytoeffector conditions. Autoclaved fungal cells were lethal for photoheterotrophic cells, but induced in photomixotrophic cells the formation of volatile n-alkanes, phthalides, coumarins, and elemicine. Most of the compounds elicited reached a concentration maximum between 20 and 30 h after addition of the mycelium, whereas the group of n-alkanes increased steadily during the 90 h monitored. Maximum concentrations were: 12 mg of graveolone, 1 mg of bergapten, 0.5 mg of sedanenolide, and 0.5 mg of n-tetradecane per 1 nutrient medium. A dose/effect relationship was found; 10 to 25 g of fungal wet weight per 1 culture medium resulted in maximum accumulation of volatiles. The formation of volatiles by photomixotrophic in vitro cells is discussed as an integral part of plant responses to ecological stress.     

F?nge von Habichten (Accipiter gentilis) im Wurzacher Ried: Kritische Fragen zu einem beh?rdlich genehmigten Wiedereinbürgerungsprojekt

Zusammenfassung Während des unter der Trägerschaft des Landesjagdverbandes Baden-Württemberg e. V. seit 1978 laufenden Projektes zur Wiedereinbürgerung des Birkhuhnes im Wurzacher Ried wurde behördlicherseits der Wegfang von Habichten unter bestimmten Auflagen genehmigt. Nach Beringung mußten die Vögel wieder freigelassen werden. Eine vergleichende Untersuchung der von den Projektbetreibern veröffentlichten Berichte und der Beringungsdokumente ergab erhebliche Ungereimtheiten. Die aus den Berichten der Projektbetreiber zu entnehmende Zahl von 168 gefangenen, angeblich beringten und angeblich wieder freigelassenen Habichten steht der Zahl von 98 in den Beringungslisten dokumentierten Vögel gegenüber. Bereits ein Vergleich zwischen der Wiederfundrate der dokumentierten Vögel (1 %) mit der Fänglingfundrate der Vogelwarte Radolfzell (13 %) ergab einen hochsignifikanten Unterschied. Auch der Vergleich mit der Fundrate (15 %) einer anderen, parallel laufenden Verfrachtungsstudie zeigte, daß die Wiederfundrate der Wurzacher Habichte hochsignifikant geringer ist. Die Darstellung der Projektbetreiber, wonach 1978–1981 insgesamt 90 Habichte durch die Landesanstalt für Umweltschutz verfrachtet worden sein sollen, wurde durch die Landesanstalt nicht bestätigt. Die Landesanstalt bestätigte nur fünf verfrachtete Habichte aus dem Wurzacher Projekt. Obwohl der Habichtfang im März nicht erlaubt war, haben die Projektbetreiber insgesamt vier Habichte im März gefangen und zumindest einen (nach eigenen Angaben jedoch alle!) auch noch verfrachtet. Vermutlich im Auftrag von Geflügelzüchtern wurden sieben Habichte, darunter vier Altvögel, innerhalb eines kleinen Gebietes in einer Entfernung von 13–18,5 km vom Wurzacher Ried gefangen. So besteht begründeter Verdacht, daß ein beträchtlicher Teil der im oder um das Wurzacher Ried gefangenen Habichte nicht wieder freigelassen, sondern getötet wurde. Eine Reihe der in den Projektberichten enthaltenen Angaben lassen zudem ganz erhebliche Zweifel an der Fachkompetenz der Projektbetreiber aufkommen.

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Goshawk (Accipiter gentilis) trapping in the nature reserve Wurzacher Ried: critical questions on a Black Grouse (Tetrao tetrix) reintroduction project

Summary A Black Grouse reintroduction project was established in 1978 in the nature reserve Wurzacher Ried in southwestern Germany. Since the beginning of this project the staff had a licence to catch, ring and to release but not to kill goshawks. A comparison of the very low Euring-recovery-rate of 1 percent from this project with the Euring-recovery-rate of the Vogelwarte Radolfzell (13 %) showed a significant difference (p=0.00017). In contrast, we found no difference between the recovery-rate (15 %) of a study with almost similar design conducted by the University of Tübingen (Schmidt-Koenig 1982) only 100 km apart from the Wurzacher Ried and the Euring-recovery-rate of the Vogelwarte Radolfzell (p=0.624). The recovery-rate of the reintroduction project differed, however, significantly from the recovery-rate ofSchmidt-Koenig (p=0.006). Furthermore, we found serious inconsistencies between the project reports and the ringing documents concerning this project. Our results indicate, that most of the goshawks caught in the project Wurzacher Ried may not have been released but illegally killed by the project staff.

     

Glycosylinositol-phosphoceramide in the free-living protozoan Paramecium primaurelia: modification of core glycans by mannosyl phosphate.

Glycolipids synthesized in a cell-free system prepared from the free-living protozoan Paramecium primaurelia and labelled with [3H]mannose and [3H]glucosamine using GDP-[3H]mannose and UDP-[3H]N-acetyl glucosamine, respectively, were identified and structurally characterized as glycosylinositol-phosphoceramides (GIP-ceramides). The ceramide-based lipid was also found in the GIP membrane anchor of the G surface antigen of P.primaurelia, strain 156. Using a combination of in vitro labelling with GDP-[3H]mannose and in vivo labelling with 33P, we found that the core glycans of the P.primaurelia GIP-ceramides were substituted with an acid-labile modification identified as mannosyl phosphate. The modification of the glycosylinositol-phospholipid core glycan by mannosyl phosphate has not been described to date in other organisms. The biosynthesis of GIP-ceramide intermediates in P.primaurelia was studied by a pulse-chase analysis. Their structural characterization is reported. We propose the following structure for the putative GIP-ceramide membrane anchor precursor of P.primaurelia surface proteins: ethanolamine phosphate-6Man-alpha 1-2Man-alpha 1-6Man-(mannosyl phosphate)-alpha 1-4glucosamine-inositol-phosphoceramide.     

Rate-controlling steps of oxidative phosphorylation in rat liver mitochondria. A synoptic approach of model and experiment

The contribution of different steps to the control of oxidative phosphorylation in isolated rat liver mitochondria was investigated by a combination of experiments and computer simulations. The parameters of the mathematical model of phosphorylating mitochondria were derived from experimental data. The model correctly describes the competition between ATP utilization inside and outside mitochondria for the ATP generated in mitochondria. On the basis of the good agreement between experiments and simulations, the contribution of different steps to the control of respiration was estimated by computing their control strengths, i.e., the influence of their activities on the rate of respiration. The rate-controlling influences vary depending on the load of oxidative phosphorylation. The predominant steps are: in the fully active state (State 3) — the hydrogen supply to the respiratory chain; in the resting state (State 4) — the proton leak of the mitochondrial inner membrane; in states of non-maximum ATP export — the adenine nucleotide translocator. Titrations of respiration with phenylsuccinate, antimycin, oligomycin and carboxyatractyloside completely support these conclusions.