Changes in the ultrastructure of mitochondria of roots ofTriticale subjected to anaerobiosis

Summary Anaerobiosis induces the formation of tubular inclusions within the mitochondria ofTriticale roots. These inclusions show a close relationship to the inner membrane of the mitochondrial envelope. Mitochondria of anaerobically cultured roots also become larger and fewer in number. Cristae become less numerous and more dilated in appearance. Despite these alterations the mitochondria retain their metabolic integrity, if the period of anaerobiosis does not exceed 24 hours. Periods of anaerobiosis in excess to 24 hours are increasingly deleterious to the metabolism and structure of the root cells and will eventually lead to their death.     

Inhibition of sugar uptake in adenosine-treated 3T3 cells

Addition of 5 to 250 micromolar adenosine to the culture medium resulted in a 30–80% inhibition of the rate of uptake of 2-deoxyglucose or 3–0-methylglucose by sparse or confluent 3T3 cells within three hours. The inhibition of deoxyglucose uptake could be reversed partially by changing the cells to medium without adenosine for two hours and could be prevented completely by the addition of persantin, an inhibitor of nucleoside uptake. The adenosine effect is not due to inhibition of pyrimidine synthesis, since it is not prevented by uridine. It is not seen in 3T6 cells lacking adenosine kinase. The inhibition could be observed on confluent cells whose deoxyglucose uptake was stimulated by insulin, epidermal growth factor (EGF), calf serum or calcium phosphate. Although the percentage stimulation over control by these factors varied, the percentage inhibition by addition of adenosine of the stimulated rates, as well as the unstimulated rate, was relatively constant. EGF, insulin and calcium phosphate caused little or no stimulation of deoxyglucose uptake by sparse cells, whether adenosine treated or untreated. The results suggest that adenosine acts intracellularly after phosphorylation to regulate sugar uptake through a mechanism which is independent of the regulation by hormones and cell density.     

Dihydroisocoumarins and phthalide from wood samples infested by fungi

Wood samples, infested by fungi during storage, were shown to contain, besides the known 5-methyl-mellein, additional (3R)-8-hydroxy-3-methyl-3,4-dihydroisocoumarins substituted by 7-methyl, 5-formyl, 5-carboxy, 5-hydroxy, 5-methoxy, 6-methoxy-5-methyl and 6,7-dimethoxy-5-methyl groups, as well as 6-formyl-7-hydroxy-5-methoxy-4-methylphthalide. Several 2-methylchromanones were synthesized in order to show that this class of compounds can be distinguished from 3-methyl-3,4-dihydroisocoumarins by MS.     

Mucronulatol,mucroquinone and mucronucarpan,isoflavonoids from Machaerium mucronulatum and M. villosum

Additionally to the cinnamylphenols described in a previous paper, wood samples of Machaerium mucronulatum and M. villosum contain isoflavones, besides (?)-duartin, (?)- and (±)-mucronulatol [(3S)- and rac-7,3′-dihydroxy-2′,4′-dimethoxyisoflavan], (?)-mucroquinone [(3S)-2-methoxy-5-(7-hydroxy-8-methoxychroman-3-yl)-1,4-benzoquinone] and (+)-mucronucarpan [(6aS,11aS)-2,10-dihydroxy-3,9-dimethoxypterocarpan]. The constitutions of mucronulatol, mucroquinone and mucronucarpan were deduced by spectra and degradations, and confirmed by syntheses.     

Changes in the Microvasculature and Interstitium of Aged Human Mandibles and Maxillae1

The present study describes the age changes to the microvasculature and connective tissue interstitium of the osteons and periosteums of aged human mandibles and maxillae. The mandibles and maxillae obtained from 14 and 19 year old males, respectively, were also studied. In the nutrient canals of the aged osteons, the walls of the arterioles and venules stained intensely PAS positive, and alcian blue negative. The walls of the blood capillaries were thick and strongly PAS positive. There was a deposition of PAS positive material in the connective tissue stroma of the nutrient canals which progressed to the obliteration of the canal space. Many of the nutrient canals exhibited diffuse calcification within the connective tissue interstitium localized around the blood vessels. The lacunae and canaliculi of those osteons in which the nutrient canals were partially or completely obliterated were filled with PAS material. None of these histochemical changes were seen in the osteons of young individuals. The microvasculature of the aged periosteum showed similar changes. The periosteal tissue consisted of thick collagenous bundles and few osteogenic cells. There was a thin darkly stained amorphous calcified layer forming the bone surface.     

Comparison of structural requirements for interaction of the same peptide with I-Ek and I-Ed molecules in the activation of MHC class II-restricted T cells.

We have analyzed the interaction of the hen egg-white lysozyme (HEL) peptide 107-116 with the MHC class II molecule I-Ek, using truncated and single residue substitution analogues to measure activation of I-Ek-restricted, 107-116-specific T cell hybridomas and competition for Ag presentation by I-Ek molecules. These results have been compared with previous findings on the interaction of the same peptide with the I-Ed molecule. Stimulation of T cell hybridomas by truncated peptides defines the sequence 108-116 as the minimum epitope necessary for activation of both I-Ek- and I-Ed-restricted T cell hybridomas. Substitution analysis pinpoints three residues (V109, A110, and K116) in the sequence 108-116 as being critical for binding to I-Ek molecules and demonstrates the involvement of most other residues in recognition by T cells. Results previously obtained for binding of HEL 107-116 to I-Ed molecules indicated that peptide residues R112, R114, and K116 were critical for interaction with I-Ed. Comparison of these results indicates a difference in the likely MHC contact residues between the HEL sequence 108-116 and I-Ed or I-Ek molecules, suggesting that the same HEL peptide assumes a different conformation in the binding site of these two MHC molecules. This in turn affects residues interacting with the specific T cell receptor. According to the hypothetical tridimensional structure predicted for class II molecules, the difference in MHC contact residues observed within the sequence 108-116 can be related to polymorphic amino acids in the binding site of I-Ek and I-Ed molecules. A search through published binding data for a common pattern in this and other I-Ek-binding peptides has permitted us to derive a possible motif for predicting peptide binding to I-Ek molecules. This putative motif was tested by determining binding to I-Ek of an unbiased panel of about 150 synthetic peptides. Binding data indeed demonstrate the presence of this motif in the majority of good binders to I-Ek molecules.     

Pollination and Breeding System of Vellozia squamata (Liliales: Velloziaceae): A Species of the Brazilian Cerrados

The pollination biology and breeding system of Vellozia squamata (Velloziaceae), a species of cerrado vegetation in Central Brazil, were studied. V. squamata is unusual in being pollinated by a few, generalist bee species despite having very large flowers, and having a distinctive pulsed flowering phenology. The species is self-incompatible but with a late-acting, post-fertilization rejection mechanism.     

Occurrence of macro B chromosomes in Astyanax scabripinnis paranae (Pisces,Characiformes, Characidae)

B chromosomes occur in several Neotropical fish species. Cytogenetic analysis of 27 specimens (15 females and 12 males) of Astyanax scabripinnis paranae from the Araquá river (a small headwater tributary of the Tietê river) shows that this population has 2n=50 chromosomes (4M+30 SM+4ST+12A), two chromosome pairs with NORs and conspicuous C-band positive blocks in the terminal position of the long arm of four chromosome pairs. In this population, eight females presented 2n=51 chromosomes and the extra chromosome was a large metacentric similar in size and morphology to the first chromosome pair in the karotype. This accessory chromosome is entirely heterochromatic in C-banded metaphases and shows a late replication pattern evidenced by BrdU incorporation. There was no significant correlation between the presence of B chromosomes and increased NOR activity at the P>0.05 level. Some aspects related to these B chromosomes are discussed.     

Epilithic diatom community response to years of P04 fertilization: Kuparuk River,Alaska (68 N Lat.)

An arctic river was fertilized continuously through the ice-free season with phosphoric acid beginning in 1983. The epilithic diatom community increased in biomass in the first two years in response to the added limiting nutrient (Peterson et al., 1983). The diatom community switched from one dominated by Hannea arcus to one dominated by species of Achnanthes and Cymbella. The immediate responses to the P-addition were decreases in both the Shannon diversity and evenness indices. By the second year, the community diversity increased downriver reaching maximal species richness (110–127 spp). In 1985–1987, the epilithic algal biomass decreased an order of magnitude with both whole-river PO₄ (1985, 1987) and PO₄ + NH₄ addition (1986). In the 5th summer of fertilization, the reduction in biomass was clearly caused by a numerical increase of grazing, refugia-building chironomids (Orthocladiinae, primarily) (Gibeau, 1991; Gibeau, Miller, Hershey, in prep.). We assume the algal biomass reduction in the 3rd and 4th years was similarly caused by grazers with a two year time lag in the numerical response of these monovoltine species. The evenness of the community increased in 1986 as if it might have been grazed; however the number of immigrants was reduced. The community became dominated by Eunotia, Cymbella and Achnanthes, species either fast growing or more prostrate, as the erect species of Hannea Diatoma, and Fragillaria declined. A detrended correspondence analysis of the temporal and spatial diatom samples in species space (186 spp.) showed that the largest variation in the community was between years and less variation was associated with river fertilization. Samples from bioassay tubes run by Peterson et al. (1983) in the Kuparuk River showed P and N + P limitation as found in the river in 1983–84. Like the river samples, the largest change in the diatom community occurred between 15 and 25 day samples, more than that induced by fertilization. Diatoms sampled from all treatments taken at day 25 were more similar to one another than those sampled at day 15. Diatoms colonizing glass slides used in the bioassay tubes were dominated by Achnanthes linearis and Cymbella minuta. Of the 84 species found in bioassays, 26 species were present in all river samples for 4 years. Differences in the communities discriminated by multivariate methods were cause by changes in rare species and abundance patterns of common species.     

Coordinate pretranslational control of cAMP-dependent protein kinase subunit expression during development in the water mold Blastocladiella emersonii.

The aquatic fungus Blastocladiella emersonii provides a system for studying the regulation of expression of regulatory (R) and catalytic (C) subunits of cAMP-dependent protein kinase (PKA). Blastocladiella cells contain a single PKA with properties very similar to type II kinases of mammalian tissues. During development cAMP-dependent protein kinase activity and its associated cAMP-binding activity change drastically. We have previously shown that the increase in cAMP-binding activity during sporulation is due to de novo synthesis of R subunit and to an increase in the translatable mRNA coding for R (Marques et al., Eur. J. Biochem. 178, 803, 1989). In the present work we have continued these studies to investigate the mechanism by which the changes in the level of kinase activity take place. The C subunit of Blastocladiella has been purified; antiserum has been raised against it and used to determine amounts of C subunit throughout the fungus' life cycle. A sharp increase in C subunit content occurs during sporulation and peaks at the zoospore stage. Northern blot analyses, using Blastocladiella C and R cDNA probes, have shown that the levels of C and R mRNAs parallel their intracellular protein concentrations. These results indicate a coordinate pretranslational control for C and R subunit expression during differentiation in Blastocladiella.