Simultaneous saccharification and fermentation of straw to ethanol using the thermotolerant yeast strain Kluyveromyces marxianus imb3

The thermotolerant, ethanol-producing yeast strain Kluyveromyces marxianus IMB3 was grown at 45°C on media containing 2, 4 and 6 % (w/v) pulverized barley straw and supplemented with 2% (v/v) cellulase. Maximum ethanol concentrations produced were 2, 3 and 3.6g/l, respectively. When the pulverized straw was replaced by NaOH pretreated straw (at 2, 4 and 6% (w/v); based on original untreated straw), ethanol concentrations increased to maxima of 3.9, 8, and 12g/l, respectively. The ethanol yields amount to 20g ethanol from 100g of straw.     

DNA SYNTHESIS IN THE OOPLASM OF DROSOPHILA MELANOGASTER

Tritiated thymidine was injected into 2-day-old Drosophila melanogaster females, and tissue sections were prepared from the ovary for radioautography with both the light and electron microscopes. Besides the expected incorporation of H³-thymidine into nuclei of nurse cells and follicle cells, there was a relatively high level of incorporation of label into ooplasmic DNA. The highest level of incorporation occurred at stage 12. At the same time, the 15 nurse cell nuclei also incorporate thymidine in spite of the fact that they are breaking down and degenerating. The label in the ooplasm is not removed by extraction with DNase (although this removes nuclear label) unless extraction is preceded by a treatment with protease. Electron microscopic radioautography revealed that 36% of the silver grains resulting from decay of H³-thymidine are found over mitochondria, with a further 28% being located within 0.25 µ of these organelles. The remaining 36% of the silver grains was not found to be associated with any organelles, and it probably represents synthesis in the cytoplasm by the "storage DNA" characteristic of many eggs. It is suggested that one mechanism acting throughout the egg chamber is responsible for the synchronous synthesis of DNA in the degenerating nurse cells, in the mitochondria of the egg, and in the "storage DNA" of the ooplasm.     

Ultrastructural observations on oogenesis in Drosophila

The ultrastructure of the follicle cells and oocyte periplasm is described during the stages of oogenesis immediately prior to, during, and immediately subsequent to, vitellogenesis. A number of features have not been described previously in Drosophila. Some yolk appears prior to pinocytosis of blood proteins. However, most of the protein yolk forms while the periplasm is filled with micropinocytotic invaginations and tubules derived from the oolemma. These tubules retain the internal layer of material characteristic of coated vesicles and are found to fuse with yolk spheres. No accumulation of electron-dense material in the endoplasmic reticulum or Golgi of the oocyte is found. Both trypan blue and ferritin are accumulated by the oocyte. The follicle cells have an elaborate endoplasmic reticulum during the period of maximum yolk accumulation. Adjacent cells are joined at their base by a zonula adhaerens, forming a band around the cells, and by plaques of gap junctions. Gap junctions are also present between nurse cells and follicle cells. During chorion formation, septate junctions also appear between follicle cells, adjacent to the zonula adhaerens.     

Drosophila bearing the ocelliless mutation underproduce two major chorion proteins both of which map near this gene.

Two bands of putative Drosophila chorion mRNA, E3 and E4, have been shown to hybridize in situ near 7E11 (Spradling and Mahowald, 1979) within a region known to contain a gene, ocelliless, which may be involved in chorion production (Johnson and King, 1974). We have investigated the synthesis of “chorion mRNAs” and chorion proteins in flies carrying this mutation. A reduction of labeling of both E3 and E4 was observed in stage 12 egg chambers from homozygous ocelliless females. In addition, they produce mature oocytes which contain greatly reduced amounts of several major chorion proteins, including those normally produced in stage 12, c36 and c38. To investigate whether the reduction was due to a direct effect of the mutation, the genes for these proteins were mapped. Recombination analysis using electrophoretic variants of c36 and c38 showed that both proteins are coded on the X chromosome at a site between crossveinless and vermillion. Further mapping with the deficiency chromosomes Df(1)KA14 and Df(1)RA2 narrowed the region containing the structural genes to a 16 band region between 7E10 and 8A4. The ocelliless gene, as well as the site of in situ hybridization, are located within this same interval. In normal ovarian follicles, both c36 and c38 are produced in equal amounts and with the same developmental specificity (Waring and Mahowald, 1979). In the mutant, both are reduced to a similar extent. In oc⁺/oc heterozygotes, both the c36 and c38 genes on the mutant chromosome produce much less product than the corresponding genes on the oc⁺ chromosome. The cis-acting nature of the ocelliless mutation suggests that it may disrupt sequences involved in controlling the expression of the structural information for these proteins.     

Loss of centrioles and polyploidization in follicle cells of Drosophila melanogaster

Centrioles of the nurse cells of Drosophila have been shown to move into the oocyte prior to polyploidization of the nurse cells. In order to determine whether or not centriolar loss always occurs in polyploid insect cells, the follicular epithelium of the Drosophila ovary was studied. The DNA content of the cells was determined by cytophotometry of Feulgen-stained squash preparations. The first two endomitotic replications occur at stage 7 and 8. Two additional replications occur prior to stage 11, but the DNA content appears to be under-replicated. Centrioles are found in follicle cells until stage 10 at which time they are no longer present. At the inception of polyploidization the centrioles are no longer closely associated with each other or the nuclear envelope. Instead, they are located adjacent to the plasma membrane at the basal surface. These results closely parallel the previous results found for the nurse cells. Hence, it may be a general observation that centrioles are gradually lost in polyploid insect cells.     

Sleeping Dreams, Waking Hallucinations, and the Central Nervous System

Consciousness is now considered a primary function and activity of the brain itself. If so, consciousness is simply the brain's interpretation and integration of all the information made available to it at any given time. On the assumption that the brain is active across all states of being (wakefulness, REM sleep, and NREM sleep), this article proposes that dreaming and hallucinations represent variations on the same theme. Under usual circumstances during wakefulness, the brain ignores internally generated activity and attends to environmental sensory stimulation. During sleep, dreaming occurs because the brain attends to endogenously generated activity. In unusual settings, such as sleep-deprivation, sensory deprivation, or medication or drug ingestion, the brain attends to exogenous and endogenous activities simultaneously, resulting in hallucinations, or wakeful dreaming. This concept is supported by numerous neurologic conditions and syndromes that are associated with hallucinations.     

Evolutionary relatedness of some primate models of Plasmodium

Primate--and, specifically, monkey--malaria infections are commonly used for understanding the pathology of and immune response to the human disease because they are thought to resemble most closely the host-parasite relationship found in humans. Plasmodium cynomolgi is used extensively as a model for the human parasite, P. vivax, and P. knowlesi is used primarily as a model for the development of erythrocytic-stage vaccines. Both of these simian parasites can naturally infect man, resulting in mildly symptomatic episodes of the disease. The phylogenetic relationship between these two simian parasites and previously characterized Plasmodium species, including P. vivax, was examined by comparison of the asexually expressed small- subunit ribosomal RNA genes. Our analysis confirmed that P. vivax is most closely related to P. cynomolgi and that it remains an appropriate model of the human pathogen. Furthermore, with P. knowlesi and P. fragile, these two species form a group of closely related species, distant from other Plasmodium species. What is considered to be the most ancient of the human malaria pathogens, P. malariae, was also included in the analysis and does not group at all with other simian or human parasites.      

Evolution and phylogenetic information content of the ITS-1 region in the tiger beetle Cicindela dorsalis

Sequence divergence in the internal transcribed spacer region 1 (ITS-1) of the ribosomal DNA locus was assessed in subspecies of the coastal North American tiger beetle, Cicindela dorsalis. The spacer region was amplified using the polymerase chain reaction and cloned for sequencing. Of a total of 50 clones obtained from 12 specimens, 42 clones were different in at least one nucleotide position. In a parsimony analysis of these sequences, the main phylogenetic distinction was found to separate sequences from the Gulf of Mexico and the Atlantic Ocean. Within these two assemblages phylogenetic resolution was low, and the variation within individuals was almost as high as the variation within the entire lineage. The pattern of sequence variation suggests the existence of two forms of the ITS-1 that are maintained on different chromosomes. Polymorphisms of limited geographical distribution could be detected, and 41 additional clones were partly sequenced, to assess the geographic distribution of these polymorphisms in more detail. In a population aggregation analysis, the geographic pattern of ITS-1 distribution was basically congruent with that obtained in earlier studies from mitochondrial DNA in the same C. dorsalis populations.      

Genetic Analysis of Two Female-Sterile Loci Affecting Eggshell Integrity and Embryonic Pattern Formation in Drosophila Melanogaster

We have analyzed female-sterile mutations at the X-linked loci fs(1)Nas and fs(1)ph which show allele-specific effects on egg shell structure and embryonic pattern formation. The majority of mutant alleles at both loci lead to a collapsed egg phenotype. The maternal effect lethal phenotype is characterized by cuticle defects resembling those found in three autosomal mutants of the terminal class. We have analyzed the complementation behavior of various heteroallelic combinations at both loci and show that one such combination at the fs(1)Nas locus is capable of restoring normal fertility. We have investigated possible interactions between fs(1)Nas and fs(1)ph and also between the terminal allele of fs(1)Nas and various maternal effect mutations altering the anteroposterior polarity of embryos. We have isolated one new allele of fs(1)Nas which combines the locus-typical phenotypic features with novel cuticle phenotypes. Our results suggest that the products of fs(1)Nas and fs(1)ph are required for the stability of the vitelline membrane and are also involved in a morphogenetic pathway necessary for the correct differentiation of the terminal regions of the embryo. Possible mechanisms to account for the association of these two functions are discussed.     

Germ-line sex determination in Drosophila melanogaster

In Drosophila melanogaster, the mechanism of sex determination is substantially different in the germ line and in the soma. In the germ line, the process is not completely cell-autonomous, but requires some signals from the soma. Only some of the genes involved in somatic sex determination are also needed for germ cell development. Recent genetic studies have identified loci required for germ-line sex determination.