In situ accumulation of 3 beta-hydroxylanost-8-en-32-aldehyde in hepatocyte cultures. A putative regulator of 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity

Biphasic modulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) has been demonstrated in primary hepatocyte cultures treated with the lanosterol 14 alpha-methyl demethylase inhibitor miconazole. At concentrations of the drug which lead to suppressed levels of reductase activity, the appearance of a polar, mevalonate-derived sterol is noted. Cochromatography of the identified sterol with 3 beta-hydroxylanost-8-en-32-aldehyde tentatively identified the metabolite as a lanosterol 14 alpha-methyl demethylation intermediate. Subsequent isolation and characterization of the metabolite by gas chromatography/mass spectroscopy confirmed this structural assignment. When the lanosterol 14 alpha-methyl demethylase-deficient mutant, AR45, was treated with authentic metabolite, a suppression of HMG-CoA reductase was observed. These results demonstrate that metabolism of the oxygenated biosynthetic intermediate is not required to suppress reductase activity. The results also strongly support the hypothesis that oxygenated 14 alpha-methyl demethylase intermediates are endogenously generated modulators of HMG-CoA reductase activity.     

The emergence of a new chlorophytan system,and Dr. Kornmann's contribution thereto

In traditional chlorophytan systems the organizational level was the primary character for the distinction of main groups (classes and orders). For instance, in Fott (1971), the flagellate level corresponds with the Volvocales, the coccoid level with the Chlorococcales, the filamentous level with the Ulotrichales, the siphonocladous level with the Siphonocladales, and the siphonous level with the Bryopsidales. The new system presented here is an elaboration and emendation of recently proposed taxonomies and their underlying phylogenetic hypotheses, and it is mainly based on ultrastructural features which have become available over the last 15 years. The following criteria are used for the distinction of classes and orders: (1) architecture of the flagellate cell (flagellate cells are considered as the depositories of primitive characters); (2) type of mitosis-cytokinesis; (3) place of meiosis in the life history and, consequently, the sexual life history type; (4) organizational level and thallus architecture; (5) habitat type (marine versus feshwater and terrestrial); (6) chloroplast type. The following classes are presented: Prasinophyceae, Chlamydophyceae, Ulvophyceae (orders Codiolales, Ulvales, Cladophorales, Bryopsidales, Dasycladales), Pleurastrophyceae (?), Chlorophyceae s.s. (orders Cylindrocapsales, Oedogoniales, Chaetophorales), Zygnematophyceae, Trentepohliophyceae, Charophyceae (orders Klebsormidiales, Coleochaetales, Charales). The new system no longer reflects the traditional hypothesis of a stepwise evolutionary progression of organizational levels in which the flagellate level represents the most primitive lineage, the coccoid and sarcinoid levels lineages of intermediate derivation, and the filamentous, siphonocladous and siphonous levels the most derived lineages. Instead, it is now hypothesized that these levels have arisen over and over again in different chlorophytan lineages which are primarily characterized by their type of flagellate cell. The flagellate green algal classes Prasinophyceae (with organic body scales) and Chlamydophyceae probably represent bundles of highly conservative lineages that diverged very long ago. Consequently, extant genera and species in these classes can be expected to have emerged long ago. Fossil evidence points to a minimum age of 600 Ma of certain extant Prasinophycean genera, and molecular evidence to a minimum age of 400–500 Ma of a fewChlamydomonas species. On the contrary, the most derived “green algal” lineage, the Angiosperms, can be expected to consist of, on average, much younger genera and species. Fossil evidence points to a minimum age of genera of 5–60 Ma. Lineages of intermediate evolutionary derivation (Ulvophyceae, Chlorophyceae, Charophyceae) can be expected to encompass genera and species of intermediate age. Fossil and (limited) molecular evidence point to a minimum age of 230–70 Ma of extant genera in Bryopsidales, Dasycladales and Cladophorales (Ulvophyceae) and of 250–80 Ma of extant genera in Charales (Charophyceae).     

A comparative description of mitochondrial DNA differentiation in selected avian and other vertebrate genera

Levels of mitochondrial DNA (mtDNA) sequence divergence between species within each of several avian (Anas, Aythya, Dendroica, Melospiza, and Zonotrichia) and nonavian (Lepomis and Hyla) vertebrate genera were compared. An analysis of digestion profiles generated by 13-18 restriction endonucleases indicates little overlap in magnitude of mtDNA divergence for the avian versus nonavian taxa examined. In 55 interspecific comparisons among the avian congeners, the fraction of identical fragment lengths (F) ranged from 0.26 to 0.96 (F = 0.46), and, given certain assumptions, these translate into estimates of nucleotide sequence divergence (p) ranging from 0.007 to 0.088; in 46 comparisons among the fish and amphibian congeners, F values ranged from 0.00 to 0.36 (F = 0.09), yielding estimates of P greater than 0.070. The small mtDNA distances among avian congeners are associated with protein-electrophoretic distances (D values) less than approximately 0.2, while the mtDNA distances among assayed fish and amphibian congeners are associated with D values usually greater than 0.4. Since the conservative pattern of protein differentiation previously reported for many avian versus nonavian taxa now appears to be paralleled by a conservative pattern of mtDNA divergence, it seems increasingly likely that many avian species have shared more recent common ancestors than have their nonavian taxonomic counterparts. However, estimates of avian divergence times derived from mtDNA- and protein-calibrated clocks cannot readily be reconciled with some published dates based on limited fossil remains. If the earlier paleontological interpretations are valid, then protein and mtDNA evolution must be somewhat decelerated in birds. The empirical and conceptual issues raised by these findings are highly analogous to those in the long-standing debate about rates of molecular evolution and times of separation of ancestral hominids from African apes.      

Methods for computing the standard errors of branching points in an evolutionary tree and their application to molecular data from humans and apes

Statistical methods for computing the standard errors of the branching points of an evolutionary tree are developed. These methods are for the unweighted pair-group method-determined (UPGMA) trees reconstructed from molecular data such as amino acid sequences, nucleotide sequences, restriction-sites data, and electrophoretic distances. They were applied to data for the human, chimpanzee, gorilla, orangutan, and gibbon species. Among the four different sets of data used, DNA sequences for an 895-nucleotide segment of mitochondrial DNA (Brown et al. 1982) gave the most reliable tree, whereas electrophoretic data (Bruce and Ayala 1979) gave the least reliable one. The DNA sequence data suggested that the chimpanzee is the closest and that the gorilla is the next closest to the human species. The orangutan and gibbon are more distantly related to man than is the gorilla. This topology of the tree is in agreement with that for the tree obtained from chromosomal studies and DNA-hybridization experiments. However, the difference between the branching point for the human and the chimpanzee species and that for the gorilla species and the human-chimpanzee group is not statistically significant. In addition to this analysis, various factors that affect the accuracy of an estimated tree are discussed.      

Sulfur dynamics in mineral horizons of two northern hardwood soils. A column study with 35S

Sulfur dynamics of two Spodosols were ascertained using soil columns constructed from homogenized mineral soil from nothern hardwood ecosystems at the Huntington Forest (HF) in the Adirondack Mountains of New York and Bear Brook Watershed in Maine (BBWM). Columns were leached for 20 weeks with a simulated throughfall solution with³⁵SO₄ ^2-. Sulfur constituents were similar to those of other Spodosols, with the organic S fractions (C-bonded S and ester sulfate) constituting over 90% of total S. HF soil columns had higher total S (14.9 mol S g^-1) than that for the BBWM soil columns (7.4 mol g^-1) primarily due to higher C-bonded S in the former.Initially, adsorbed SO₄ ^- accounted for 5 and 4% of total S for the BBWM and HF soil columns, respectively. After 20 weeks, adsorbed SO₄ ^2- decreased (81%) in BBWM and increased (33%) in HF soil columns. For both HF and BBWM soil columns, C-bonded S increased and ester sulfate decreased, but only for HF columns was there a net mineralization of organic S (5.6% of total S). The greatest decrease in ester sulfate occurred at the top of the columns.Leaching of³⁵S was less than 0.5% of the³⁵S added due to its retention in various S constituents. There was an exponential decrease in³⁵S with column depth and most of the radioisotope was found in C-bonded S (70–88 and 70–91% for BBWM and HF, respectively). The rapid turnover of adsorbed SO^2- ₄ was reflected in its high specific activity (834 and 26 kBq mol^-1 S for BBWM and HF, respectively). The lower specific activity of adsorbed SO₄ ^2- in HF was attributable to greater isotopic dilution by non-radioactive SO^2- ₄ derived from greater organic S mineralization in the HF versus the BBWM columns.Both soil columns initially had high levels of NO^- ₃ which resulted in the generation of H⁺ and net retention of SO₄ ^2- in the early phase of the experiment due to pH dependent sulfate adsorption; later NO₃ ^- decreased and SO₄ ^2- was desorbed. Leaching of NIO₃ ^- and SO₄ ^2- was correlated with losses of Mg²⁺ and Ca²⁺ of which the latter was the dominant cation.Analyses using both S mass balances and radioisotopes corroborate that for BBWM soil columns, SO^2- ₄ adsorption-desorption dominated the S biogeochemistry while in HF soil columns, organic S mineralization-immobilization processes were more important. It is suggested that similar techniques can be applied to soils in the field to ascertain the relative importances of SO₄ ^2- adsorption processes and organic S dynamics.     

The role of predators and parasites in controlling populations ofEarias insulana,heliothis armigera andBemisia tabaci on cotton in the Syrian Arab Republic

Studies on the relationship between insect pests and their predators and parasites were conducted in the Syrian cotton agro-ecosystem from 1980 up to 1983.Earias insulana (Boisduval) was found to be the most damaging pest, whileHeliothis armigera (Hübner) was not an annual returning problem. Among the entomophagous insects found, coccinellids were most numerous during the months June and July and the hemipterous predators were more abundant during August and September. When predator numbers were reduced with insecticide applications, a significant increase in the bollworm populations occurred, resulting in significant reductions in seed cotton yields. Parasites were relatively not important for the control of lepidopterous pests on cotton. Egg and larva populations ofE. insulana andH. armigera were parasitized byTrichogramma spp. andHabrobracon brevicornis Wesmael respectively. Populations ofBemisia tabaci (Gennadius) were heavily parasitized byEretmocerus mundus Mercet.      

Anonymous nuclear DNA markers in the American oyster and their implications for the heterozygote deficiency phenomenon in marine bivalves

A puzzling population-genetic phenomenon widely reported in allozyme surveys of marine bivalves is the occurrence of heterozygote deficits relative to Hardy-Weinberg expectations. Possible explanations for this pattern are categorized with respect to whether the effects should be confined to protein-level assays or are genomically pervasive and expected to be registered in both protein- and DNA-level assays. Anonymous nuclear DNA markers from the American oyster were employed to reexamine the phenomenon. In assays based on the polymerase chain reaction (PCR), two DNA-level processes were encountered that can lead to artifactual genotypic scorings: (a) differential amplification of alleles at a target locus and (b) amplification from multiple paralogous loci. We describe symptoms of these complications and prescribe methods that should generally help to ameliorate them. When artifactual scorings at two anonymous DNA loci in the American oyster were corrected, Hardy-Weinberg deviations registered in preliminary population assays decreased to nonsignificant values. Implications of these findings for the heterozygote-deficit phenomenon in marine bivalves, and for the general development and use of PCR-based assays, are discussed.