Annual cycle of stored spermatozoa within the ovaries of Helicolenus dactylopterus dactylopterus (Teleostei, Scorpaenidae)

The aim of this work was to analyse the ultrastructure of storage crypts and stored spermatozoa, and to describe changes during the annual reproductive cycle of the bluemouth Helicolenus dactylopterus dactylopterus , which has internal fertilization and a zygoparous mode of reproduction. Spermatozoa had elongated heads and long midpieces, two characteristics which are thought to be fairly advanced and correlated with internal fertilization, as is the case of the bluemouth. A remarkable spermatozoon feature was the retention of a significant quantity of cytoplasm around the head, a condition that appeared to be related to nourishment during the long storage period, up to 10 months in the intraovarian crystal structures of the female. Male sex cells' protection against the female immune system was ensured by junctional complexes between the crypt cells composed of tight junctions and desmosomes.     

Altered protein-chromophore interaction in dicyclohexylcarbodiimide-modified purple membrane sheets

Previous studies of N,N'-dicyclohexylcarbodiimide (DCCD)-modified bacteriorhodopsin (Renthal, R. et al. (1985) Biochemistry 24, 4275-4279) used reaction conditions (detergent micelles) that are not optimal for subsequent physical studies. The present work describes new conditions for reaction of bacteriorhodopsin with DCCD in intact purple membrane sheets in the presence of 4.5% (v/v) diethylether and light. Like the detergent reaction system, the reaction is light induced, incorporates approximately 1 mol [14C]DCCD per mol bacteriorhodospin, and results in a bleached chromophore. Peptide mapping indicates that the likely site of modification in intact membranes is identical to the site in the detergent reaction system: Asp 115. The retinal chromophore of DCCD-modified purple membrane has an absorbance maximum at 390 nm and very little induced circular dichroism. The retinal is easily extracted in hexane, yielding a 3:1 ratio of all-trans to 13-cis retinal. Borohydride reduces the retinal onto the protein within the 1-71 region of the amino acid sequence. These results suggest that Asp-115 is near the retinal binding cavity of bacteriorhodopsin. When DCCD reacts with Asp 115, retinal is displaced from its binding site.     

Insulin-like growth factor-I supports proliferation of autocrine thymic lymphoma cells with a pre-T cell phenotype

We have studied the phenotypic characteristics and growth properties of murine T lymphoma cell lines derived from primary x-ray-induced thymic lymphomas at the earliest stage at which they can be detected, and well before spreading to other organs has occurred. These cell lines serve as model systems for the earliest events in T cell lymphoma induction, before tumor cell progression and spreading to other organs. We find that primary x-ray-induced T cell lymphoma lines have phenotypic characteristics of thymic pre-T cells and show no proliferative response to any of the IL tested nor to other hematopoietic growth factors. However, they do proliferate in response to insulin-like growth factor I (IGF-I) and to a small autocrine peptide distinct from IGF-I, which we term lymphoma growth factor. One of the earliest lesions in T cell lymphoma induction may therefore be an inhibition of differentiation at one of several specific points. In its early stages, T lymphoma cell growth may be restricted to an environment where local concentrations of specific growth factors such as IGF-I or lymphoma growth factor are sufficiently high.     

Excess amino acid polymorphism in mitochondrial DNA: contrasts among genes from Drosophila, mice, and humans

Recent studies of mitochondrial DNA (mtDNA) variation in mammals and Drosophila have shown an excess of amino acid variation within species (replacement polymorphism) relative to the number of silent and replacement differences fixed between species. To examine further this pattern of nonneutral mtDNA evolution, we present sequence data for the ND3 and ND5 genes from 59 lines of Drosophila melanogaster and 29 lines of D. simulans. Of interest are the frequency spectra of silent and replacement polymorphisms, and potential variation among genes and taxa in the departures from neutral expectations. The Drosophila ND3 and ND5 data show no significant excess of replacement polymorphism using the McDonald-Kreitman test. These data are in contrast to significant departures from neutrality for the ND3 gene in mammals and other genes in Drosophila mtDNA (cytochrome b and ATPase 6). Pooled across genes, however, both Drosophila and human mtDNA show very significant excesses of amino acid polymorphism. Silent polymorphisms at ND5 show a significantly higher variance in frequency than replacement polymorphisms, and the latter show a significant skew toward low frequencies (Tajima's D = -1.954). These patterns are interpreted in light of the nearly neutral theory where mildly deleterious amino acid haplotypes are observed as ephemeral variants within species but do not contribute to divergence. The patterns of polymorphism and divergence at charge-altering amino acid sites are presented for the Drosophila ND5 gene to examine the evolution of functionally distinct mutations. Excess charge-altering polymorphism is observed at the carboxyl terminal and excess charge-altering divergence is detected at the amino terminal. While the mildly deleterious model fits as a net effect in the evolution of nonrecombining mitochondrial genomes, these data suggest that opposing evolutionary pressures may act on different regions of mitochondrial genes and genomes.      

Inhibition of CI857-controlled recombinant gene expression in Escherichia coli at very low concentrations of glucose

Summary The effects of glucose on biomass and temperature-mediated recombinant gene expression has been studied in Escherichia coli batch cultures. Glucose, above 2 g/L, has inhibitory effects on aerobic cell growth and specially on gene expression. Although lower concentrations have no influences on biomass, a tenuous, but still remarkable concentration-dependent reduction of gene expression levels was observed even at about 0.1 g/L.     

Molecular characterization of the proteinase-encoding gene, prb1, related to mycoparasitism by Trichoderma harzianum

The soil fungus Trichoderma harzianum is a mycoparasitic fungus known for its use as a biocontrol agent of phytopathogenic fungi. Among other factors, Trichoderma produces a series of antibiotics and fungal cell wall-degrading enzymes. These enzymes are believed to play an important role in mycoparasitism. Among the hydrolytic enzymes, we have identified a basic proteinase (Prb1) which is induced by either autoclaved mycelia, fungal cell wall preparation or chitin; however, the induction does not occur in the presence of glucose. The proteinase was purified and biochemically characterized as a serine proteinase of 31 kDa and pl 9.2. Based on the sequence of three internal peptides, synthetic oligonudeotide probes were designed. These probes allowed subsequent isolation of a cDNA and its corresponding genomic clone. The deduced amino acid sequence indicates that the proteinase is synthesized as a pre-proenzyme and allows its classification as a serine proteinase. Northen analysis shows that the induction of this enzyme is due to an increase in the corresponding mRNA level.     

Spawning of Basilichthys microlepidotus (Jenyns)

Characteristics of the reproductive cycle of Basilichthys microlepidotus (Jenyns), an atherinid fish living in a South American Mediterranean arid zone, are analysed. Specimens 9-170 mm in total length were captured from August 1982 to April 1984 in the Rio Choapa, in northern Chile (31%%45' S, 71°15' W). Analysis of gonadosomatic index fluctuations, and determination of the proportion of mature males and females captured during the study period, allowed the estimation of the duration of the reproductive cycle.
Females 70–170 mm in total length contained 2000–9000 intraovarian oocytes, of which approximately 80% were vitellogenic (100–1300 μm in diameter) and exhibited a polymodal distribution. A further 4 to 15% of the total oocytes, measured 1700–2000 μm in diameter and appeared ready to be spawned. In 30% of the examined ovaries some of the largest oocytes exhibited signs of degeneration, or atresia. Juveniles approximately 30 mm in total length were captured over most of the study period. Results strongly suggest that B. microlepidotus has multiple spawnings.     

Effect of norepinephrine and vasopressin on renal kallikrein excretion in rats

The purpose of this study was to investigate the effect of norepinephrine and vasopressin on urinary kallikrein excretion in the rat. Two studies were undertaken: (a) acute experiments in which the rats were infused with 30% dextrose in water with the addition of norepinephrine or vasopressin, (b) chronic experiments in which the drugs were infused during seven days through an osmotic minipump. In acute experiments, urinary kallikrein excretion increased without modification in urinary flow and glomerular filtration rate. In chronic experiments, urinary kallikrein excretion was not modified in norepinephrine-treated rats and decreased in vasopressin-infused animals. This decrease followed the modifications of the urine flow. In chronic experiments the dextrose infusion increased urinary kallikrein excretion. In all the groups studied a positive correlation between urine flow and urinary kallikrein excretion was observed. It is concluded that norepinephrine and vasopressin are important stimulators of the urinary kallikrein excretion only in those circumstances where it is necessary to eliminate an excess of water.