Physical, chemical and morphologic dimensions of human hepatitis A virus strain CR326 (38578).

CR326 human hepatitis A virus purified by isopycnic banding from infected marmoset sera was shown to consist of 27 nm spherical particles on electron microscopic examination. The particles were identified as hepatitis A virus by tests for infectivity and by specific neutralization of infectivity with convalescent human hepatitis A serum. Also, indentical 27 nm viruses in liver extracts gave specific reactions with hepatitis A antisera when tested by immune electron microscopy. The bouyant density of the virus in CsCl was 1.34 and it was heat (60 degrees), ether and acid stable but was destroyed by heat (100 degrees), formalin (1:4000) and ultraviolet irradiation. Electron microscopic studies of sections of infected marmoset liver showed intracytoplasmic virus particles, usually in vesicles. Presumptive findings for RNA, together with the intracytoplasmic location of the virus, indicated the virus to be of RNA-type. The attributes of the virus indicate it is closely related to the enterovirus family and not to hepatitis B virus.     

Haterozygosity and human evolution

Effects of heterozygosity are well known in the world of plants and animals as a symptom of high hybrid vigour appearing, for example, as greater body size or a modified level of metabolism. Two questions arise: whether effects of heterozygosity occur also in the species Homo sapiens, and if so whether it appears in man in a way similar to that in animals.These questions have been the subject of our investigations during the past 10 years. The present paper aims at showing what the effects of heterozygosity in man consist of and what significance it may have for the processes of evolution.     

Long-term observations of currents on the central Great Barrier Reef continental shelf

Time-series are presented of wind, sea levels, currents and temperature in the central region of the Great Barrier Reef continental shelf over the period 1980 to 1982 (inclusive). Except for occasional periods of calm weather or during the passage of tropical cyclones, the wind was alternately southwestward or northwestward, with periods of several days to several weeks. The fluctuations of the low-frequency sea level, longshore current and wind components, but not temperature, were highly coherent over distances >400 km longshore and over the shelf width. The temperature and the atmospheric pressure controlled much of the seasonal changes in the sea level. A simple model for wind-driven circulation using a non-linear bottom friction law is able to account for most of the variance of the currents with perios <20 days. An additional poleward base flow was present at all sites, probably driven by the circulation in the Coral Sea. There were also large interannual variations, possibly associated with the El Niño Southern Oscillation phenomena. These observations imply that the advection and dispersion over the Great Barrier Reef of fish eggs and larvae, coral planulae etc., and hence reef recruitment, experience considerable variability at time scales of days of years.     

Transport mechanisms and the potential movement of planktonic larvae in the central region of the Great Barrier Reef

It is suggested that considerable inter-reef dispersal of reef fishes and many benthic invertebrates is likely in the central region of the Great Barrier Reef. Larvae are most abundant in spring-summer when currents on the outer shelf, where most of the coral reefs occur, are almost entirely unidirectional and southeastward (longshore). Net drift on the outer shelf at this time is likely to be greater, but the dispersion smaller, than that nearshore at the same time due to more extensive periodic reversals of water movement in the latter area than the former. Net drift on the outer shelf in winter will be significantly more restricted, but the dispersion greater, than in summer due to extensive periodic reversals of currents in this area during the trade wind (winter) season. These conclusions suggest that reefs within the Central Great Barrier Reef are biologically interconnected and interdependent; a result of considerable significance for management of reefs within the Great Barrier Reef marine park.Australian Institute of Marine Science Contribution No. 250     

The influence of wetlands, decaying organic matter, and stirring by wildlife on the dissolved oxygen concentration in eutrophicated water holes in the Seronera River, Serengeti National Park, Tanzania

The dissolved oxygen concentration (DO) was sampled during a diurnal cycle in three water holes heavily used by wildlife and with distinctive biological features along the Seronera River. The DO fluctuated widely (by up to 11.5 mg l⁻¹) as a function of time, mechanical stirring and aeration by animals, and the presence of fringing wetlands. The DO cycle was successfully modeled (within 0.3 mg l⁻¹) by assuming that the four dominant processes were photosynthesis and respiration by algae near the surface, trapping by wetlands, decomposition of dead organic matter on the bottom, and stirring/aeration by hippos. The rate of DO decline from the decay of dead organic matter was equal to the rate of DO removal by algal respiration at night.     

Characterization of a soluble stable human cytomegalovirus protease and inhibition by M-site peptide mimics.

The human cytomegalovirus (HCMV) protease is a potential target for antiviral chemotherapeutics; however, autoprocessing at internal sites, particularly at positions 143 and 209, hinders the production of large quantities of stable enzyme for either screening or structural studies. Using peptides encompassing the sequence of the natural M-site substrate (P5-P5', GVVNA/SCRLA), we previously demonstrated that substitution of glycine for valine at the P3 position in the substrate abrogates processing by the recombinant protease in vitro. We now demonstrate that introduction of the V-to-G substitution in the P3 positions of the two major internal processing sites, positions 143 and 209, in the mature HCMV protease renders the enzyme stable to autoprocessing. When expressed in Escherichia coli, the doubly substituted protease was produced almost exclusively as the 30-kDa full-length protein. The full-length V141G, V207G (V-to-G changes at positions 141 and 207) protease was purified as a soluble protein by a simple two-step procedure, ammonium sulfate precipitation followed by DEAE ion-exchange chromatography, resulting in 10 to 15 mg of greater than 95% pure enzyme per liter. The stabilized enzyme was characterized kinetically and was indistinguishable from the wild-type recombinant protease, exhibiting Km and catalytic constant values of 0.578 mM and 13.18/min, respectively, for the maturation site (M-site) peptide substrate, GVVNASCRLARR (underlined residues indicate additions to or substitutions from peptides derived from the wild-type substrate). This enzyme was also used to perform inhibition studies with a series of truncated and/or substituted maturation site peptides. Short nonsubstrate M-site-derived peptides were demonstrated to be competitive inhibitors of cleavage in vitro, and these analyses defined amino acids VVNA, P4 through P1 in the substrate, as the minimal substrate binding and recognition sequence for the HCMV protease.     

Stress-resistant Translation of Cathepsin L mRNA in Breast Cancer Progression

The cysteine protease cathepsin L (CTSL) is often thought to act as a tumor promoter by enhancing tumor progression and metastasis. This goes along with increased CTSL activity in various tumor entities; however, the mechanisms leading to high CTSL levels are incompletely understood. With the help of the polyoma middle T oncogene driven breast cancer mouse model expressing a human CTSL genomic transgene, we show that CTSL indeed promotes breast cancer metastasis to the lung. During tumor formation and progression high expression levels of CTSL are maintained by enduring translation of CTSL mRNA. Interestingly, human breast cancer specimens expressed the same pattern of 5′ untranslated region (UTR) splice variants as the transgenic mice and the human cancer cell line MDA-MB 321. By polyribosome profiling of tumor tissues and human breast cancer cells, we observe an intrinsic resistance of CTSL to stress-induced shutdown of translation. This ability can be attributed to all 5′ UTR variants of CTSL and is not dependent on a previously described internal ribosomal entry site motif. In conclusion, we provide in vivo functional evidence for overexpressed CTSL as a promoter of lung metastasis, whereas high CTSL levels are maintained during tumor progression due to stress-resistant mRNA translation.