Characterization of divalent cation localization in the minor groove of the A(n)T(n) and T(n)A(n) DNA sequence elements by (1)H NMR spectroscopy and manganese(II)

The localization of Mn(2+) in A-tract DNA has been studied by (1)H NMR spectroscopy using a series of self-complementary dodecamer oligonucleotides that contain the sequence motifs A(n)(n) and T(n)A(n), where n = 2, 3, or 4. Mn(2+) localization in the minor groove is observed for all the sequences that have been studied, with the position and degree of localization being highly sequence-dependent. The site most favored for Mn(2+) localization in the minor groove is near the 5'-most ApA step for both the T(n)A(n) and the A(n)T(n) series. For the T(n)A(n) series, this results in two closely spaced symmetry-related Mn(2+) localization sites near the center of each duplex, while for the A(n)T(n) series, the two symmetry-related sites are separated by as much as one half-helical turn. The degree of Mn(2+) localization in the minor groove of the T(n)A(n) series decreases substantially as the AT sequence element is shortened from T(4)A(4) to T(2)A(2). The A(n)T(n) series also exhibits length-dependent Mn(2+) localization; however, the degree of minor groove occupancy by Mn(2+) is significantly less than that observed for the T(n)A(n) series. For both A(n)T(n) and T(n)A(n) sequences, the 3'-most AH2 resonance is the least broadened of the AH2 resonances. This is consistent with the observation that the minor groove of A-tract DNA narrows in the 5' to 3' direction, apparently becoming too narrow after two base pairs for the entry of a fully hydrated divalent cation. The results that are reported illustrate the delicate interplay that exists between DNA nucleotide sequence, minor groove width, and divalent cation localization. The proposed role of cation localization in helical axis bending by A-tracts is also discussed.     

The populations of Pseudophoxinus zeregi (Heckel) in Israel and Syria and the status of Pseudophoxinus (Pararhodeus) drusensis (Pellegrin) (Pisces:Cyprinidae)

Pseudophoxinus zeregi (Heckel) and P. (Pararhodeus) drusensis (Pellegrin) have been examined from populations near their respective type localities. It was found that there is no basis for the subgenus Pararhodeus and that the two forms are best regarded as subspecies of Pseudophoxinus zeregi.     

Seasonal changes in the circadian activity rhythm of light-dark (LD) and completely dark (DD) regimen in the mouse submandibular gland in the presence of light-dark (LD) and completely dark (DD) regimen

Influence of seasons on circadian activity changes and the influence of one and six weeks of DD upon these changes of acid phosphatase (AP) and beta-acetylglucosaminidase (AM) was studied in the submandibular gland of sexually mature male mice. Total enzyme activity was determined in tissue homogenates at four-hour intervals in March, June, October, and February under standard LD12/12 conditions and after one and six weeks of the DD regime. The rhythms were analysed according to cosinor method. Under constant lighting conditions the seasonal differences in the AM circadian activity rhythm were found. AP activity was considerably less influenced by seasonal changes. Both enzyme activity changes were independent of each other and each rhythm was differently influenced by DD. In the case of AM the most pronounced circadian activity changes had the highest amplitude and mesor occurred in summer. The strongest influence of DD upon this enzyme activity rhythm was observed in spring and summer especially after the first week, after six weeks the acrophase returned to the LD group value (spring). In autumn and winter the reaction to DD was different to that of summer and spring. For AP the circadian changes of activity were non-rhythmic in spring, whereas in all other seasons the acrophases occurred almost at the same time in the afternoon. In DD the activity rhythm significantly changed after six weeks. In all seasons, except spring the circadian rhythm of activity was not observed after six weeks of DD. An attempt was made to explain the observed results by the certain kind of genetic memory present in laboratory animals the neurohormonal system of which is influenced by seasonal changes.     

Metabolism of fluoroacetate in the skink (Tiliqua rugosa) and the rat (Rattus norvegicus)

Administration of 100 mg sodium fluoroacetate (compound 1080) per kilogram body weight to T. rugosa resulted in a 3.4-fold increase in plasma citrate levels 48 h after dosing while administration of 3 mg sodium fluoroacetate per kilogram body weight to R. norvegicus produced a fivefold increase in plasma citrate levels within 4 h. Administration of 300 mg sodium fluoroacetate per kilogram body weight reduced the oxygen consumption of the skink by between 2.5 and 11% while in the rat, 2 mg sodium fluoroacetate per kilogram body weight reduced oxygen consumption by between 28 and 57%. Aconitate hydratase activity in extracts of liver acetone powders from T. rugosa was less inhibited by (-)erythrofluorocitrate (Ki: 0.065 mM) than that in extracts derived from R. norvegicus (Ki: 0.026 mM). The rate of defluorination of fluoroacetate in erythrocytes and in extracts of liver acetone powders of T. rugosa was 8- and 4.5-fold greater, respectively, than that found in similar preparations from R. norvegicus. A rapid rate of defluorination together with a low reliance on aerobic respiration favoured detoxification of fluoroacetate in T. rugosa rather than its conversion into fluorocitrate. Though defluorination in this species helped to minimize the immediate effects of fluoroacetate on aerobic respiration, it resulted in rapid depletion of liver glutathione levels.     

Ecology and phenology of the Dysderidae (Araneae) in the Djurdjura National Park (Algeria)

The ecology and phenology of the Dysderidae (Araneae) were investigated in two different sites (Tala-Guilef and Tikjda), located at opposite slopes of the Djurdjura National Park in Algeria. The following questions were investigated: Are there any endemic species in the studied area? Do the slopes' orientation influence the abundance, richness and diversity of these spider communities? Are the activity and the vital cycle of the three most abundant species different in the two different plant physiognomies (cedar forests and alpine grasslands)? The Dysderidae were collected monthly for 2 years using pitfall traps. In total, 1532 specimens were collected: 333 males, 389 females and 381 juveniles in Tala-Guilef and 230 males, 66 females and 133 juveniles in Tikjda. Among the total of 10 species sampled, three species belonging to three genera in Tala-Guilef, 10 species belonging to four genera in Tikdja and four endemic species was recorded. The slope orientation influence the abundance and the species richness. The activity and the vital cycle of the three most abundant species were not different in the cedar forests and alpine grasslands. For these species, no relationship was found between the activity cycle and the vegetation physiognomy.     

Ultrastructure and Sporogony of Eimeria (syn. Epieimeria) anguillae (Apicomplexa) in the Eel (Anguilla anguilla)

ABSTRACT. Ultrastructural studies on Eimeria (syn. Epieimeria ) anguillae (Apicomplexa), parasite of the digestive tract of the eel, have shown that the development of this parasite takes place completely within the host cell. Merogony and gamogony are intracellular but in the epicytoplasmic position. Sporogony is also located within the epithelial cells, which agrees with assignment of this coccidian in the family Eimeriidae. However, depending on the intensity of infection and the physiopathological reaction of the host, the gamont may behave in two ways. 1) In massive infections, gamogony stages cause a genuine destruction of intestinal epithelium. Large numbers of gamonts form nodules and parts of the seriously destroyed epithelium peel off and are released into the lumen of the gut and quickly discharged into the outer environment. This discharged epithelium envelops cells containing immature oocysts that then sporulate outside the host. 2) In light infections, the host cells, which are necrotic due to the presence of a zygote, are pushed between the surrounding intact cells towards the base of the epithelium. Closely above its basal lamella, the oocyst then undergoes sporulation. These results show no taxonomically important biological features (e.g. special mode of implantation to the host cell or active movement of the zygote). Because the morphological characteristics of Epieimeria do not differ significantly from Eimeria , we propose to suppress the genus Epieimeria Dyková and Lom, 1981, and relegate its species into the genus Eimeria .     

Convergence of the cyst in cysticercoids (Cestoda,Hymenolepidata) and the tegument in cystacanths (Acanthocephala)

It was shown that despite fundamental differences in the organization of the tegument of cystacanths and the cyst wall of cysticeroids, they have a number of features similar in localization, morphology, terms of formation, and functional value. These include a powerful outer layer of glycocalyx on the surface of the tegument, filling of the distal department with dense substance, and the presence of complicated fibrillar systems. It is supposed that the nature of these particularities is convergent and is determined by the similarity of the life cycles of Hymenolepidata and Acanthocephala.     

Determination and Evaluation of the Metals and Metalloids in the Chapeu-de-couro (Echinodorus macrophyllus (Kunth) Micheli)

The Chapeu-de-couro (Echinodorus macrophyllus (Kunth) Micheli) is a native plant from Brazil, which has been mainly used in medicinal application being a potent antirheumatic and diuretic, in the production of soft drinks, and also in the ornamentation of aquariums. In this paper, the metals and metalloids for the leaves of chapeu-de-couro collected in the Paraguacu River from the city Cachoeira, Bahia State, Brazil, was determined and evaluated using multivariate analysis. The samples were digested using nitric acid and hydrogen peroxide and were analyzed using inductively coupled plasma mass spectrometry. The accuracy of the method was confirmed by analysis of a certified reference material of apple leaves, furnished by National Institute of Standard and Technology. The study involved 15 samples of the Paraguacu River. The results expressed as milligrams of element per kilogram of sample demonstrated that the concentration ranges varied: 1.39–5.27 for chromium, 44.85–165.39 for manganese, 0.55–0.84 for arsenic, 0.01–3.94 for antimony, and 0.18–0.31 for lead. The principal component analysis and hierarchical cluster analysis evidenced that the concentrations of the metals and metalloids varied according with the variations in the water of the Paraguacu.     

Dioecism of the genusCallicarpa (Verbenaceae) in the Bonin (Ogasawara) Islands

The morphology and function of the flowers of three species of the genusCallicarpa, C. glabra, C. nisnimurae andC. subpubescens, endemic to the Bonin Islands, revealed that all three species were dioecious. Male plants had short-styled flowers with sterile ovaries, while female plants had long-styled flowers with non-germinating inaperturate pollen grains. Such dioecism has never been reported from the family Verbenaceae. This curious sexual system with non-germinating pollen grains may have been maintained as a reward to insect pollinators.     

Investigations on the triiodothyronine (T3)-specificity of thyrotropic (TSH) and gonadotropic (HCG) hormone in the unicellular Tetrahymena

In a previous experiment thyrotropin (TSH) increased the triiodothyronine (T3) production of Tetrahymena and chorionic gonadotropin (HCG) moderately overlapped the effect. At present the production of three amino acid type (histamine, serotonin, epinephrine) and one peptide (endorphin) hormones were studied under the effect of TSH or HCG, in tryptone-yeast (TY) or salt (Losina-Losinsky) medium. The duration of the effect was 10 min. TSH significantly (with almost 20%) decreased epinephrine production in TY medium and HCG similarly decreased epinephrine and increased histamine level. In salt solution TSH as well as HCG decreased the level of serotonin. The results show that at this low level of phylogeny TSH effect is not completely thyroxine-specific, however it is not general. HCG overlaps TSH effect on epinephrine and serotonin production, however its effect is broader. The experiments also demonstrate that the effect of pituitary trop-hormones can be bidirectional in Tetrahymena, as histamine level was increased and epinephrine level was decreased by HCG, in the same cells.     

Early ossification in the skeleton of the sucker (Catostomus macrocheilus) and the guppy (Poecilia reticulata)

Catostomus macrocheilus, a sucker, and Poecilia reticulata, a live-bearer, are subjected to dissimilar mechanical stresses while most of their bony structures are forming. Osteogenesis occurs in the sucker when it is a free-living larva, whereas in the livebearer it occurs intrafollicularly. The first-formed bones ofboth species are those that meet functional demands and are subjected to the greatest stresses. These are the movable bones associated with the acts of respiration and feeding, and some of the axial bones. The cranial roof is the last to calcify. The primary differences in osteogenesis of the two species, such as in the vertebral column and caudal fin, can be correlated with differences in stress. Surface forces on calcifiable tissue perhapsassist in the deposition of calcium salts.     

Mitochondria-rich cells in the astigmatic mites,Diplaegidia columbae (Buchholz) (Analgidae) and Falculifer rostratus (Buchholz) (Falculiferidae) (Acari: Astigmata)

Mitochondria are well-characterized intracellular organelles usually concentrated in locations of high energy consumption. Light microscopic and transmission electron microscopic observations of the internal anatomy of the feather mites Diplaegidia columbae and Falculifer rostratus were conducted. In the anterior half of the bodies of the mites, we found several dozen of distinctive mitochondria-rich (MR) cells filled with abundant, large mitochondria. Mitochondria are placed individually or enclosed in small groups within an elaborated lamellar system forming a mitochondria–lamellae complex (MLC). The role of the MLC as well as the MR cells is not clear at present, but their involvement in heat generation is hypothesized and briefly discussed.     

Deletion of the alpha(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep

Nuclear transfer offers a cell-based route for producing precise genetic modifications in a range of animal species. Using sheep, we report reproducible targeted gene deletion at two independent loci in fetal fibro-blasts. Vital regions were deleted from the alpha(1,3)galactosyl transferase (GGTA1) gene, which may account for the hyperacute rejection of xenografted organs, and from the prion protein (PrP) gene, which is directly associated with spongiform encephalopathies in humans and animals. Reconstructed embryos were prepared using cultures of targeted or nontargeted donor cells. Eight pregnancies were maintained to term and four PrP-/+ lambs were born. Although three of these perished soon after birth, one survived for 12 days. These data show that lambs carrying targeted gene deletions can be generated by nuclear transfer.     

Different sperm number in the spermatophores of Orchesella villosa (Geoffroy) (Entomobryidae) and Allacma fusca (L.) (Sminthuridae)

Members of the basal hexapod group Collembola perform indirect sperm transfer; males deposit spermatophores on the soil and these are then picked up by females for insemination. Orchesella villosa exhibits a dissociated mating behavior, while Allacma fusca has either associated mating behavior or, more commonly, produces spermatophores without pairing. Sperm ultrastructure in the two species has been studied by TEM and SEM observations to estimate the volume of these cells. The sperm number into each spermatophore has been determined by counting the fluorescent nuclei after Hoechst staining of sperm droplets squash preparations. Sperm droplets at the apex of spermatophores contain numerous spermatozoa of typical appearance, wound up and with a long anterior appendage (peduncle). Sperm droplets of Orchesella villosa are smaller (about 10 times) than in A. fusca, however they contain substantially more sperm cells (about three times). These differences could be explained as follows: (a) sperm of O. villosa are shorter than those of A. fusca (58 and 107 μm, respectively) and thus the volume of a single sperm of O. villosa is about 10 times smaller than in A. fusca; (b) considering the volume of sperm droplets and of sperm cells, the estimated number of sperm cells would be higher in A. fusca than in O. villosa; and (c) the conspicuous reduction of the sperm cells observed in A. fusca is likely linked to the aberrant meiosis present in this species, as well as in all Symphypleona studied so far, that causes a loss of 50% of their sperm.     

Factors Controlling the Bioaccessibility of Arsenic(V) and Lead(II) in Soil

The relative oral bioaccessibility of labile Pb(II) and As(V) added to soils was investigated in a well-characterized soil using a physiologically based extraction test (PBET) to simulate metal solubility in a child's digestive system. The effect of soil and PBET (i.e., simulated stomach and small intestine) pH, soil metal concentration, soil to solution ratio, and soil-metal aging time were investigated. Arsenic bioaccessibility was relatively unaffected by a variation in simulated stomach and small intestine pH over the range 2 to 7 and soil pH over the range 4.5 to 9.4. In contrast, Pb(II) bioaccessibility was strongly dependent on both the simulated stomach, small intestine, and soil pH, showing enhanced sequestration and decreased bioaccessibility at higher pH values in all cases. Although the bioaccessibility of Pb(II) was constant over the concentration range of approximately 10 to 10,000?mg/kg, the As(V) bioaccessibility significantly increased over this concentration range. The bioaccessibility of both arsenic and lead increased as the soil-to-solution ratio decreased from 1:40 to 1:100. Additional lead sequestration was not observed during 6 months of soil aging, but As(V) bioaccessibility decreased significantly during this period.     

Over-representation of the disease associated (CAG) and (CGG) repeats in the human genome.

Expansion of trimer repeats has recently been described as a new type of human mutation. Of the 64 possible trimer compositions, only the CGG and CAG repeats have been implicated in genetic diseases. This study intends to address two questions: (1) What makes the CGG and CAG repeats unique? (2) Could other trimer repeats be involved in this type of mutation? By computer analysis of trimer and hexamer frequency distributions in approximately 10 Mb of human DNA, twenty trimer motifs (ten complementary pairs) have been identified that are the most likely to be expanded. The frequency distribution study also indicated that the expanded trimer motif in Fragile-X syndrome is GGC instead of CGG. DNA linguistics studies revealed that the GGC/GCC and CAG/CTG repeats were over-represented in the human genome. Further analysis of base composition suggested that the CCA/TGG repeats may be involved in the trimer expansion mutation since they possessed many similar characteristics to GGC/GCC and CAG/CTG. The computer aided sequence analysis studies reported here may help to understand the molecular mechanisms of trimer repeat expansion.