Zinc content in selected tissues in streptozotocin-diabetic rats after maximal exercise

The Zn metabolism in experimental diabetic rats after maximal exercise was investigated. Forty male wistar rats were used, weighing 240±10 g at the beginning of this experiment. The animals were assigned to one of four experimental groups (n=10): control at rest (CR), control plus exercise (CE), diabetic at rest (DR), and diabetic plus exercise (DE). Experimental diabetes was produced by a single intraperitoneal injection of streptozotocin (STZ) (60 mg/kg). Thirty days after injection of streptozotocin, the animals of groups CE and DE were forced to acute exercise (swimming) until exhaustion. Glucose, rectal temperature (RT), pH, swimming time (ST), hematocrit (Hct), serum, and tissue (heart, liver, kidney, and muscle) Zn concentrations were measured. The streptozotocin treated animals used in the current experiment were diabetic. Increases in hepatic, renal muscle, and serum levels Zn at rest and after exercise until exhaustion were found in normal and diabetic rats. ST decreased (?180%) in the diabetic rat group. In conclusion, the results of the present study indicate that STZ-induced diabetes was associated with altered tissue Zn concentration, both at rest and after exercise.     

The role of ATP as a mediator in the action of iron complexes on cellular calcium homeostasis

The effects of the interaction between low molecular weight iron complexes (citrate, lactate, and ATP complexes) with ATP and proteins, on the modification of Ehrlich carcinoma cell calcium homeostasis have been studied. In that modification the ferric-ATP complex shows much higher activity than the others. Sodium ATP, by iron translocation from citrate and lactate, increases their activity. This phenomenon implicates ATP as a mediator on the cellular activity of the complexes. Proteins, particularly ferritin, appear to moderately reduce their activity, whereas glutathione and ascorbic acid, acting as lipid peroxidation-inhibitors, show only a slight reduction of the iron complex’s effects on cellular calcium uptake.     

Sialosylcholesterol Effects on Reconstitution of Microfilament and Glia Filament

Abstract: The effects of α-sialosylcholesterol (α-SC) on formation of either microfilament or glia filament of rat astrocytes were investigated using a reconstitution system. Polymerization of the depolymerized microfilament preparation that had been extracted from a crude cytoskeletal fraction of rat astrocytes, in the presence of 100 m M KCI and 10 m M MgCI₂, was suppressed in a dose-dependent manner by α-SC. α-SC inhibited polymerization of G-actin in a similar manner. The intensity of a-SC inhibition of G- actin polymerization was as great as that of microfilament polymerization, suggesting that the inhibition of microfilament polymerization by α-SC was due to the direct action of α-SC on actin, the main component of microfilament. α-SC depolymerized partly the polymerized microfilament preparation, which resembled F-actin (microfilament-like filaments). α-SC suppressed, in a dose-dependent manner, polymerization of a glia filament preparation that had been extracted from astrocyte cytoskeletons in the presence of phalloidin. An increase in the amount of added α-SC (up to 15 n M ) decreased the amount of the larger glia filament-like filaments, which were 10 nm thick and centrifuged down at 16,000 g for 30 min, and increased that of smaller ones precipitated only after centrifugation at 100,000 g for 1 h. The lower the concentration of the depolymerized glia filament extract, the greater was the inhibition by α-SC of the polymerization. α-SC repressed polymerization of vimentin, the dominant component of glia filament. Vimentin polymerization was more strongly inhibited by α-SC than polymerization of glia filament was. The findings suggested that α-SC suppressed polymerization of glia filament through a direct action on vimentin and that the glia filament-associated proteins increased its structural stability in the presence of α-SC.     

Variation in digestive performance between geographically disjunct populations of Atlantic salmon: countergradient in passage time and digestion rate

European Atlantic salmon (Salmo salar) populations inhabit rivers from northern Portugal to northern Norway across a wide spectrum of environmental variability. To address whether single physical factors might lead to genetic divergence of isolated populations, we compared the digestive performances total digestibility, relative nitrogen digestibility, passage time, and digestion rate (g dry matter · h^–1) — of northern (Scotland) and southern (Asturias, northern Spain) populations at three temperature regimes (5, 12, and 20° C). Total dry matter digestibilities increased directly with temperature but were similar for both populations at each of the three trials. Relative nitrogen digestibility did not differ between populations nor among temperature regimes. In contrast, passage time was significantly longer for low-than for high-latitude fish at both 5 and 20° C. When the percentage of food digested and the passage time were integrated as digestion rates (food digested per unit time), a significant population × temperature interaction consistent with a genotype × environment interaction was detected in addition to the population and temperature effects. This implies that not only is the digestive performance of the high-latitude population higher throughout the range of temperatures examined, but moreover the difference is reinforced at high temperatures, where the digestion rate of high-latitude fish was 1.6 times greater. Taken together, these two results provide preliminary evidence for countergradient variation in digestive rates of salmonids in response to variation in growth opportunity. The data support our previous work on the same two populations showing differences in growth rates, and underlie one of the possible mechnisms leading to more rapid growth of the high-latitude fish when both populations are reared in a common environment.     

Morphological evolution and changes in foraging behaviour of island and mainland populations of Blue Tit (Parus caeruleus) — a test of convergence and ecomorphological hypotheses

Summary We study the leg morphology and feeding postures of two subspecies of the Blue Tit (Parus caeruleus; Tenerife island and the Iberian Peninsula) and the Coal Tit (Parus ater; Iberian Peninsula). We search for evidence supporting the hypothesis of convergent evolution in morphological and ecological traits and we discuss the role of ecomorphological hypotheses as predictors of foraging differences at the intraspecific level. To overcome the problems introduced by environmental characteristics not related to locomotion and competition, we make observations under controlled situations to manage food quality and food access. We determine that the island Blue Tit has a longer tarsometatarsus, larger foot span and a more proximal insertion of the tibialis cranialis muscle (flexor of the tarsometatarsus) than the mainland Blue Tit. These morphological differences are consistent with the more frequent use of hanging and clinging head-up postures by the Iberian Blue Tit. Several ecomorphological hypotheses obtained at the interspecific level with other taxa, have proved to be of high predictive value for explaining ecological differences considering morphological evolution. The Tenerife Blue Tit and the Iberian Coal Tit clearly show close convergence in both feeding postures and leg structure, although some differences in morphology were found between these two species. Convergence in foraging methods between the island Blue Tit and the mainland Coal Tit can be explained without considering current interspecific competition as a determinant of niche space.     

Gibberellin A3 and Abscisic Acid Cause the Reorientation of Cortical Microtubules in Epicotyl Cells of the Decapitated Dwarf Pea

To determine whether or not the changes in the orientation ofmicrotubules (MTs) that are induced by GA₃ and ABA result fromchanges in the rate of epicotyl elongation caused by these hormones,we examined the effects of GA₃ and ABA on the orientation ofMTs in epidermal cells of decapitated epicotyls of the dwarfpea (Pisum sativum cv. Little Marvel), in which neither GA₃nor ABA causes changes in the rate of epicotyl elongation. Cuttings taken from GA₃-pretreated seedlings were decapitatedand treated with ABA. ABA eliminated the GA₃-induced predominanceof transverse MTs and treatment with ABA resulted in a predominanceof longitudinal MTs in the decapitated cuttings. However, ABAdid not reduce the rate of epicotyl elongation in these samples.Cuttings taken from ABA-pretreated seedlings were decapitatedand treated with GA₃. GA₃ caused the orientation of MTs to changefrom longitudinal to transverse in the decapitated cuttings.However, GA₃ had no promotive effect on elongation of theseepicotyls. The results indicate that both ABA and GA₃ have the abilityto change the orientation of MTs by mechanisms that do not involvechanges in the rate of cell elongation. (Received August 18, 1992; Accepted January 18, 1993)     

Three new HLA-G alleles and their linkage disequilibria with HLA-A

Three new allelic forms of the HLA-G DNA sequence (HLA-G*II, HLA-G*III, and HLA-G*IV) have been identified. With the HLA-G*I sequence (previously designated HLA 6.0) as a reference, HLA-G*II shows a silent (G A) mutation at the third base of codon 57, HLA-G*III bears a non-synonymous (A T), but conservative, (Thr Ser) substitution at the first base of codon 31, and HLA-G*IV shows two silent substitutions: (A T) at the third base of codon 107 and (G A) at the third base of codon 57. A rapid method of singling out each allele on genomic DNA has been developed by using polymerase chain reaction amplification followed by restriction endonuclease treatment. Also, more or less strong linkage disequilibria has been found between most HLA-A alleles and either HLA-G*I or *II, both being the most prevalent alleles in the population, with a genotypic frequency of 0.55 and 0.38, respectively; HLA-G*III is very rare and HLA-G*IV has a genotypic frequency of 0.07. An evolutive classification of HLA-A alleles results according to their association with either HLA-G*I or HLA-G*II, which does not correlate with the classical serological cross-reacting groups classification. The finding of a strong and selective A/G linkage disequilibria with most HLA-A alleles, together with the existence of less frequent random A/G associations, may suggest that there exist in different haplotypes true and varied A/G genetic distances (and not a recombinational hotspot). It may be inferred from preliminary data that in primates HLA-A/G haplotypes bearing G*II may have appeared later than those bearing G*I.The nucleotide sequence data reported in this paper have been submitted to the GenBank and EMBL nucleotide sequence databases and have been assigned the following accession numbers: EMBL-X60983 (HLA-G*II), GenBank-M99048 (HLA-G*III), and GenBank-L07784 (HLA-G*IV).The contribution to this paper by P. Morales and A. Corell is equal, and the order of authorship is arbitrary. Correspondence to: A. Arnaiz-Villena.     

A strong protein unfolding activity is associated with the binding of precursor chloroplast proteins to chloroplast envelopes

Protein conformational changes related to transport into chloroplasts have been studied. Two chimaeric proteins carrying the transit peptide of either ferredoxin or plastocyanin linked to the mouse cytosolic enzyme dihydrofolate reductase (EC 1.5.1.3.) were employed. In contrast to observations in mitochondria, we found in chloroplasts that transport of a purified ferredoxin-dihydrofolate reductase fusion protein is not blocked by the presence of methotrexate, a folate analogue that stabilizes the structural conformation of dihydrofolate reductase. It is shown that transport competence of this protein in the presence of methotrexate is not a consequence of alteration of the folding characteristics or methotrexate binding properties of dihydrofolate reductase by fusion to the ferredoxin transit peptide. Binding of dihydrofolate reductase fusion proteins to chloroplast envelopes is not inhibited by low temperature and it is only partially diminished by methotrexate. It is demonstrated that the dihydrofolate reductase fusion proteins unfold, despite the presence of methotrexate, on binding to the chloroplast envelopes. We propose the existence of a strong protein unfolding activity associated to the chloroplast envelopes.     

Influence of a Small Number of Mature T Cells in Donor Bone Marrow Inocula on Reconstitution of Lymphoid Tissues and Negative Selection of a T Cell Repertoire in the Recipient

Allo-chimerism and clonal elimination of self antigen (Ag) (Ia + Mls-1^a) reactive Vβ6+ T cells were analyzed and compared between allogeneic bone marrow (BM) chimeras reconstituted with BM cells which had been treated with anti-Thy-1 monoclonal antibody (mAb) plus complement (C) (T^– chimeras) and BM chimeras which had been reconstituted with BM cells pretreated with anti-Thy-1 mAb alone (T⁺ chimeras). When lethally irradiated AKR (Mls-1^a) mice were reconstituted with BM cells from B10 or B10 H-2 congenic mice, both T⁺ and T^– chimeras were entirely free of signs of graft-versus-host reaction (GVHR). However, complete replacement of the AKR lymphoid tissues by donor BM cells was accomplished at an early stage in T⁺ chimeras but not in T^– chimeras. On the other hand, clonal elimination of Vβ6⁺ T cells reactive to the recipient Ag (Mls-1^a) was abolished in T⁺ chimeras but successfully induced in T^– chimeras. The Vβ6⁺ T cells not eliminated in T⁺ chimeras showed depressed responses against Mls-1^a antigens. The findings herein demonstrate that T cells which contaminate a BM inoculum survive in recipient mice after treatment with anti-Thy-1 mAb without C in vitro followed by BMT. The surviving T cells have been estimated to represent fewer than 0.5% of the BM cells inoculated. These cells appear to accelerate the full replacement of recipient lymphoid tissues by donor cells. Furthermore, the T cells which survive in the marrow inoculum influence eventually the development of a tolerant state in the T cell repertoire of the donor.     

Cloning and Sequencing of Two New Verotoxin 2 Variant Genes of Escherichia coli Isolated from Cases of Human and Bovine Diarrhea

We cloned and sequenced two new Verotoxin 2 (VT2) variant genes: one from an Escherichia coli strain from a case of bovine diarrhea and the other from an E. coli strain from a patient with diarrhea. The nucleotide and amino acid sequences of these two genes were highly homologous with, but distinct from those of the VT2, VT2vha, VT2vhb, SLT-IIv (VT2vp1) and SLT-IIva (VT2vp2) genes. Their nucleotide sequences were much more closely homologous to that of VT2vh than to that of VT2vp. Search for these two new genes in other Verocytotoxin-producing E. coli strains resulted in the isolation of 2 strains carrying one of the new VT2 variant genes, one strain from Tokyo and the other from Canada.