Autophagy initiation correlates with the autophagic flux in 3D models of mesothelioma and with patient outcome

Understanding the role of autophagy in cancer has been limited by the inability to measure this dynamic process in formalin-fixed tissue. We considered that 3-dimensional models including ex vivo tumor, such as we have developed for studying mesothelioma, would provide valuable insights. Using these models, in which we could use lysosomal inhibitors to measure the autophagic flux, we sought a marker of autophagy that would be valid in formalin-fixed tumor and be used to assess the role of autophagy in patient outcome. Autophagy was studied in mesothelioma cell lines, as 2-dimensional (2D) monolayers and 3-dimensional (3D) multicellular spheroids (MCS), and in tumor from 25 chemonaive patients, both as ex vivo 3D tumor fragment spheroids (TFS) and as formalin-fixed tissue. Autophagy was evaluated as autophagic flux by detection of the accumulation of LC3 after lysosomal inhibition and as autophagy initiation by detection of ATG13 puncta. We found that autophagic flux in 3D, but not in 2D, correlated with ATG13 positivity. In each TFS, ATG13 positivity was similar to that of the original tumor. When tested in tissue microarrays of 109 chemonaive patients, higher ATG13 positivity correlated with better prognosis and provided information independent of known prognostic factors. Our results show that ATG13 is a static marker of the autophagic flux in 3D models of mesothelioma and may also reflect autophagy levels in formalin-fixed tumor. If confirmed, this marker would represent a novel prognostic factor for mesothelioma, supporting the notion that autophagy plays an important role in this cancer.     

The effects of the cold winter of 1962/63 on the helgoland population ofBranchiostoma lanceolatum (Pallas)

Summary 1. Seawater temperatures at Helgoland were subnormal for six months during the winter of 1962/63 and fell to –1.3° C. Quantities of silt from the shores and estuaries of the mainland were released into the water when pack ice around the island melted in late February.2. In a sample dredged from a ground near Helgoland in April, 1963, 40% of the lancelets consisted of isolated notochords. From comparison with a sample taken in November, 1962, it is estimated that at least 50% of the population died during the winter. The average number of animals per Van Veen grab fell from 6.9 before to 3.0 after the cold period.3. None of the autumn settlement of newly metamorphosed animals survived, and there was a very high death rate estimated at 90% among the largest animals.4. An analysis of activity in relation to temperature inBranchiostoma lanceolatum collected from Helgoland and Naples showed a temperature tolerance of 3° to 27° C in both populations with active swimming at temperatures of 10° to 20° C. There is no muscular movement below 3° C which is the lower limit of the normal winter temperature at Helgoland. It is suggested that the species is best adapted to Mediterranean conditions.5. The pharyngeal mechanism cannot operate normally below 3° C and although it is considered unlikely that starvation was the primary cause of death, other effects of a reduced pharyngeal current may have proved lethal.6. It is held that the depth to which lancelets of different sizes burrow into the substrate could account for differential mortality through cold.

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Wie Wirkungen des kalten Winters 1962/63 auf die Helgoland-Population vonBranchiostoma lanceolatum (Pallas)

Kurzfassung Über die Hälfte der HelgoländerBranchiostoma lanceolatum-Population starb, als die Wassertemperatur während des strengen Winters 1962/63 unter 0° C herabsank. Dies geht aus der Analyse von Fängen hervor, welche mit Van Veen und Hensen Dredgen gemacht worden waren. In der jüngsten Altersgruppe der Population stieg die Sterberate auf 100% an. Die Empfindlichkeit von Individuen verschiedener Größe gegenüber niedriger Temperatur wird diskutiert hinsichtlich ihres Verhaltens sowohl im Laboratorium als auch im Grobsand ihres Habitats. Untersuchungen über das Aktivitätsmuster von Vertretern der Helgoland- und der Neapel-Populationen in Abhängigkeit von verschiedenen Temperaturen machen wahrscheinlich, daßB. lanceolatum eine mediterrane Art ist, welche sich nordwärts in den Englischen Kanal und die Nordsee ausgebreiter hat. Die Tiere sind nicht in der Lage, sich bei Temperaturen unter 3° C — wie sie bei Helgoland im Winter normalerweise vorkommen — fortzubewegen.

     

Investigation into the production of bacteriostatic substances by fungi VI. Examination of the larger Basidiomycetes

The present paper, the sixth of the series, gives the results of testing some 700 species of the larger Basidiomycetes for bacteriostatic properties. Initially, the 'juice' extracted from the sporophore was tested and this was followed in certain cases by a test of the metabolism solution produced by the fungus in culture. The results indicate that the larger Basidiomycetes are among the more promising fungus groups which produce antibiotics and that they compare favourably in this respect with the Aspergilli and the Penicillia. Of the 700 species tested approximately 70 are strongly and approximately 100 weakly positive against Staphylococcus aureus and/or Bacterium colt.     

The Effect of Novel Research Activities on Long-term Survival of Temporarily Captive Steller Sea Lions (Eumetopias jubatus)

Two novel research approaches were developed to facilitate controlled access to, and long-term monitoring of, juvenile Steller sea lions for periods longer than typically afforded by traditional fieldwork. The Transient Juvenile Steller sea lion Project at the Alaska SeaLife Center facilitated nutritional, physiological, and behavioral studies on the platform of temporary captivity. Temporarily captive sea lions (TJs, n = 35) were studied, and were intraperitoneally implanted with Life History Transmitters (LHX tags) to determine causes of mortality post-release. Our goal was to evaluate the potential for long-term impacts of temporary captivity and telemetry implants on the survival of study individuals. A simple open-population Cormack-Jolly-Seber mark-recapture model was built in program MARK, incorporating resightings of uniquely branded study individuals gathered by several contributing institutions. A priori models were developed to weigh the evidence of effects of experimental treatment on survival with covariates of sex, age, capture age, cohort, and age class. We compared survival of experimental treatment to a control group of n = 27 free-ranging animals (FRs) that were sampled during capture events and immediately released. Sex has previously been show to differentially affect juvenile survival in Steller sea lions. Therefore, sex was included in all models to account for unbalanced sex ratios within the experimental group. Considerable support was identified for the effects of sex, accounting for over 71% of total weight for all a priori models with delta AICc <5, and over 91% of model weight after removal of pretending variables. Overall, most support was found for the most parsimonious model based on sex and excluding experimental treatment. Models including experimental treatment were not supported after post-hoc considerations of model selection criteria. However, given the limited sample size, alternate models including effects of experimental treatments remain possible and effects may yet become apparent in larger sample sizes.