STUDIES ON THE ORIGIN OF RIBOSOMES IN AMOEBA PROTEUS

The origin of cytoplasmic RNA and ribosomes was studied in Amoeba proteus by transplantation of a radioactive nucleus into an unlabeled cell followed by examination of the cytoplasm of the recipient for the presence of label. When a RNA-labeled nucleus was used, label appeared in the ribosomes, ribosomal RNA, and soluble RNA. Since the kinetics of appearance of labeled RNA indicates that the nucleus was not injured during the transfer, and since the transferred nuclear pool of labeled acid-soluble RNA precursors is inadequate to account for the amount of cytoplasmic RNA label, it is concluded that cytoplasmic ribosomal RNA is derived from acid-insoluble nuclear RNA and is probably transported as an intact molecule. Likewise, cytoplasmic soluble RNA probably originated in the nucleus, although labeling by terminal exchange in the cytoplasm is also possible. The results were completely different when a protein-labeled nucleus was grafted into an unlabeled host. In this case, label was found only in soluble proteins in the host cell cytoplasm, and there were no (or very few) radioactive ribosomes. This suggests that the nuclear pool of ribosomal protein and ribosomal protein precursors is relatively small and perhaps nonexistent (and, furthermore, shows that there was no cytoplasmic ribosomal contamination of the transferred nucleus).     

Incidence of Clostridium botulinum Type E in Salmon and Other Marine Fish in the Pacific Northwest

Salmon, sole, cod, oysters, clams, and crabs from ocean waters along the coast of Oregon and Washington were examined for the presence of Clostridium botulinum type E. The organism was detected by identification of the type E toxin in enrichment cultures of the viscera of individual fish. Of 369 salmon specimens, 48 yielded cultures containing toxin lethal to mice, and almost half of the toxic cultures were shown to contain botulinal toxin, chiefly type E. Eighteen of 113 sole and cod specimens, 4 of 22 Dungeness crab specimens, 5 of 16 oyster specimens, and 27 of 115 clam specimens gave rise to cultures containing botulinal toxin which was usually type E, although types A and B were occasionally encountered.     

Virus Aggregation as the Cause of the Non-neutralizable Persistent Fraction

The non-neutralizable or persistent fraction of virus populations has been found to be caused by aggregated virus. Detailed investigation was performed with the prototype strain of echovirus type 4 (Pesascek), as this virus is notorious for its large non-neutralizable fraction. When Pesascek virus was clarified by low-speed centrifugation, homologous antiserum hardly neutralized the virus. However, when the virus was filtered through membranes having a porosity only twice the diameter of the virus, monodispersed virus was obtained which was efficiently neutralized. Serum titers were up to 1,000 times higher if the neutralization test was carried out with monodispersed virus. Virus in non-neutralizable aggregates was found to constitute 30% of the infective units of unfiltered Pesascek virus but only 0.1% of the antigenically related DuToit strain. This explains why DuToit strain has been a more satisfactory indicator strain for detecting type 4 antibodies, regardless of the echo 4 strain used for inducing the antibodies. Clarified suspensions and ultrafiltrates of viruses belonging to the picorna-, reo-, myxo-, adeno-, herpes-, and poxvirus groups were studied. Clarified suspensions yielded persistent fractions of 0.005% for poliovirus, of 0.1% for reovirus, of 0.6% for influenza virus, of <0.001% for adenovirus, of 0.06% for herpesvirus, and of 10 to 30% for vaccinia virus. In all cases the persistent fractions were removed by membrane filters which had a pore diameter no larger than twice that of the virus under test, and the high concentration of virus in each ultrafiltrate was completely neutralized by antiserum.     

Electronic recording of mosquito activity

Spontaneous locomotor activity of mosquitoes (Aedes aegypti) was tested over twenty-four hour periods using an electronic recording device which gave a permanent time graph of activity. Single mosquitoes were placed on a wire grid with alternate strands connected to the positive and negative poles of an electric circuit. Each time the mosquito moved, the electric current changed and the event was recorded by a pen-writer. The number of peaks per time interval gave the index of activity. Variables which may affect activity include age, physiological state, sex and strain. A distinct activity cycle was evident in both virgin and mated females but not in males; peak activity came in the early evening and activity was lowest in the early afternoon.

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Zusammenfassung Die spontane lokomotorische Aktivität von Mücken (Aedes aegypti) wurde über 24stündige Perioden mit Hilfe einer elektronischen Registriereinrichtung untersucht, die eine ununterbrochene Zeitschreibung der Aktivität ergab. Einzelne Mücken wurden auf einen Gürtel feiner Drähte gesetzt, deren Stränge abwechselnd zu den positiven und negativen Polen eines elektrischen Stromkreises führten. Jedesmal wenn sich die Mücke bewegte, änderte sich der elektrische Stromfluß; dieses Ereignis wurde von einer Schreibfeder aufgezeichnet. Die Anzahl der Ausschläge pro Zeiteinheit ergab den Aktivitätsindex. Variable, welche die Aktivität beeinflussen, umfassen Alter, physiologischen Zustand, Geschlecht und Abstammung. Bei jungfräulichen wie bei begatteten Weibchen war ein bestimmter Aktivitätszyklus erkennbar, jedoch nicht bei Männchen; der Aktivitätsgipfel lag in den frühen Abendstunden und die Aktivität war am zeitigen Nachmittag am geringsten.

     

Expression of members of the Saccharomyces cerevisiae hsp70 multigene family

The hsp70 multigene family of Saccharomyces cerevisiae is a complex multigene family, composed of members exhibiting complex patterns of regulation. Expression of some members is induced after a heat shock, whereas expression of others is repressed. Some members of the family are expressed during exponential growth. One gene, SSA3, shows an unusual pattern of expression during approach to stationary phase. While most RNAs decrease in abundance, SSA3 RNA levels dramatically increase. The constitutive expression of SSA3 in cells lacking adenylate cyclase activity suggests that cAMP modulates SSA3 expression.     

Stress and relapse of breast cancer.

To elucidate the association between stressful life events and the development of cancer the influence of life stress on relapse in operable breast cancer was examined in matched pairs of women in a case-control study. Adverse life events and difficulties occurring during the postoperative disease free interval were recorded in 50 women who had developed their first recurrence of operable breast cancer and during equivalent follow up times in 50 women with operable breast cancer in remission. The cases and controls were matched for the main physical and pathological factors known to be prognostic in breast cancer and sociodemographic variables that influence the frequency of life events and difficulties. Severely threatening life events and difficulties were significantly associated with the first recurrence of breast cancer. The relative risk of relapse associated with severe life events was 5.67 (95% confidence interval 1.57 to 37.20), and the relative risk associated with severe difficulties was 4.75 (1.58 to 19.20). Life events and difficulties not rated as severe were not related to relapse. Experiencing a non-severe life event was associated with a relative risk of 2.0 (0.62 to 7.47), and experiencing a non-severe difficulty was associated with a relative risk of 1.13 (0.38 to 3.35). These results suggest a prognostic association between severe life stressors and recurrence of breast cancer, but a larger prospective study is needed for confirmation.     

Characterization of a Nerve Growth Factor-Stimulated Protein Kinase in PC12 Cells Which Phosphorylates Microtubule-Associated Protein 2 and pp250

Treatment of PC12 cells with nerve growth factor (NGF) resulted in the rapid, but transient, activation of a protein kinase which specifically phosphorylated an endogenous 250-kDa cytoskeletal protein (pp250). We report that the microtubule-associated protein, MAP2, is an alternative substrate for the NGF-activated kinase. NGF treatment maximally activated the kinase within 5 min; however, the activity declined with longer exposure to NGF. The enzyme was localized predominantly in microsomal and soluble fractions and phosphorylated MAP2 on serine and threonine residues. The soluble enzyme was fractionated by DEAE chromatography and gel filtration and had an apparent Mr of 45,000. The enzyme was purified to near homogeneity by chromatofocussing and had a pI of 4.9. Kinetic analysis revealed that NGF treatment caused a sevenfold increase in Vmax for MAP2. The Km with respect to the MAP2 substrate was approximately 50 nM and was not altered by NGF treatment. A novel feature of the NGF-stimulated enzyme was its sharp dependence on Mn2+ concentration. The active enzyme is likely to be phosphorylated, because inclusion of phosphatase inhibitors was required for recovery of optimal activity and the activity was lost on treatment of the enzyme with alkaline phosphatase. Histones, tubulin, casein, bovine serum albumin, and the ribosomal subunit protein S-6 were not phosphorylated by this enzyme. The NGF-stimulated kinase was distinct from A kinase, C kinase, or other NGF-stimulated kinases. The rapid and transient activation of the protein kinase upon NGF treatment suggests that the enzyme may play a role in signal transduction in PC12 cells.     

Toremifene, a new antiestrogenic compound in the treatment of metastatic mammary cancer. A phase II study

Toremifene is a new antiestrogenic compound. Toremifene has definite antitumor effect in advanced breast cancer. The response rate in the present phase II study among postmenopausal women, mostly not pretreated with systemic therapy and with ER positive or not determined ER status in tumor tissue, was 11/23 (48%; 95% confidence interval 37-59%) including 6 complete responses. The toxicity profile was similar to that of tamoxifen. It is concluded that toremifene is at least as active as tamoxifen in advanced breast cancer and that a randomized study between these two antiestrogens is indicated.