24 resultados para Kimberlite alteration
em Brock University, Canada
Resumo:
The formation of the Sar Cheshmeh porphyry Cu-Mo deposit is related to the culmination of calc-alkaline igneous activity in the Kerman region. The deposit comprises a suite of Late Cenozoic intrusive sub-volcanic and extrusive rocks emplaced into a folded series of Eocene andesitic lavas and pyroclastic sediments. The earliest stage of magmatism was emplacement of a large granodiorite stock about 29 m.y.b.p. This was followed by intrusion of two separate porphyritic bodies at 15 (Sar Cheshrneh porphyry) and 12 m.y.b.p. (Late porphyry) and a series of sub-volcanic dikes between 12 and 9 m.y.b.p. Magmatic activity terminated with multi-phase extrusion of a Pelean dacitic dome complex between 10 and 2.8 m.y.b.p. The country rocks and the earlier porphyritic intrusions are pervasively altered to biotite-rich potassium silicate (metasomatic and hydrothermal) sericite-clay, phyllic and chlorite-clay, argillic assemblages. These grade outwards to an extensive propylitic zone. Within the ore body, the later intra-. and post-mineral dikes only reach the propylitic grade. At least three different sets of quartz veins are present, including a sericite-chlorite-quartz set which locally retrogrades pervasive secondary biotite to sericite. In the hypogene zone, metasomatic and hydrothermal alteration is related to all stages of magmatism but copper mineralization and veining are restricted to a period of 15 to 9 m.y.b.p.related to the early intrusive phases. The copper mineralization and silicate alteration do not fit a simple annular ring model but have been greatly modified by, 1. The existence of an ititial, outer ring, of metasomatic alteration overprinted by an inner.ring of hydrothermal alteration and, 2. later extensive dilating effects of intra- and post-mineral dikes. The hydrothermal clay mineral assemblage in the hypogene zone is illite-chlorite-kaolinite-smectite (beidellite). Preliminary studies indicate that the amount of each of these clays varies vertically and that hydrothermal zonation of clay minerals is possible. However, these minerals alter to illite-kaolinite assemblages in the supergene sulfide zone and to more kaolinite-rich assemblages in the supergene leached zone. Hydrothermal biotite breaks down readily in the supergene zone and is not well preserved in surface outcrops. The distribution of copper minerals in the supergene sulfide enrichment zone is only partly related to rock type being more dependent on topography and the availability of fractures.
Resumo:
High chromium content in kimberlite indicator minerals such as pyrope garnet and diopside is often correlated with the presence of diamonds. In this study, kimberlite indicator minerals were examined using visible light reflectance spectroscopy to determine if chromium content can be correlated with spectral absorption features. The depth of absorption features in the visible spectral region were correlated with the molecular percentage of chromium and other first series transition metal elements obtained by electron microprobe data. In the visible part of the spectrum, chromium is evident by 3 absorption features in the pyrope reflectance spectrum; one isolated and narrow feature at the wavelength 689 nm was used to correlate with the chromium mol %. The isolation of this feature in the pyrope spectra is advantageous since it is not directly affected by other proximal absorption bands that could be caused by other transition metals. Analysis of the feature indicates that as grain volume increases the depth of the absorption feature will also increase. Clustering grain volumes into fractions yields better correlation between absorption depth and mol % chromium. Other types of garnet (almandine, grossular, spessartine) and kimberlite indicator minerals (olivine, diopside, chromite, ilmenite) were analyzed to determine if other absorption features could be used to predict the proportion of specific transition metal elements. Diopside in particular illustrates the same isolated chromium absorption feature as pyrope and may indicate mol percent but needs further study with larger sample sets.
Resumo:
Microbial ichnofossils in volcanic rocks provide a significant record of subsurface microbes and potentially extraterrestrial biosignatures. Here, the textures, mineralogy, and geochemistry of two continental basaltic hydrovolcanic deposits - Reed Rocks and Black Hills - in the Fort Rock Volcanic Field (FRVF) are investigated. Methods include petrographic microscopy, micro and powder X-ray diffraction, SEM/BSE/EDF imaging, energy dispersive spectroscopy, stable isotopes, and X-ray fluorescence. Petrographic analysis revealed granular and tubular textures with biogenic morphologies that include terminal enlargements, septate divisions, branching forms, spiral filaments, and ovoid bodies resembling endolithic microborings described in ocean basalts. They display evidence of behaviour and a geologic context expressing their relative age and syngenicity. Differences in abiotic alteration and the abundance/morphotype assemblage of putative microborings between the sites indicate that water/rock ratio, fluid composition and flux, temperature and secondary phase formation are influences on microboring formation. This study is the first report of reputed endolithic microborings in basalts erupted in a continental lacustrine setting.
Resumo:
The rock sequence of the Tertiary Beda Formation of S. W. concession 59 and 59F block in Sirte Basin of Libya has been subdivided into twelve platformal carbonate microfacies. These microfacies are dominated by muddy carbonates, such as skeletal mudstones, wackestones, and packstones with dolomites and anhydrite. Rock textures, faunal assemblages and sedimentary structures suggest shallow, clear, warm waters and low to moderate energy conditions within the depositional shelf environment. The Beda Formation represents a shallowing-upward sequence typical of lagoonal and tidal flat environments marked at the top by sabkha and brackish-water sediments. Microfossils include benthonic foraminifera, such as miliolids, Nummulites, - oerculina and other smaller benthonics, in addition to dasycladacean algae, ostracods, molluscs, echinoderms, bryozoans and charophytes. Fecal pellets and pelloids, along with the biotic allochems, contributed greatly to the composition of the various microfacies. Dolomite, where present, is finely crystalline and an early replacement product. Anhydrite occurs as nodular, chickenwire and massive textures indicating supratidal sabkha deposition. Compaction, micr it i zat ion , dolomit izat ion , recrystallization, cementation, and dissolution resulted in alteration and obliteration of primary sedimentary structures of the Beda Formation microfacies. The study area is located in the Gerad Trough which developed as a NE-SW trending extensional graben. The Gerad trough was characterized by deep-shallow water conditions throughout the deposition of the Beda Formation sediments. The study area is marked by several horsts and grabens; as a result of extent ional tectonism. The area was tectonically active throughout the Tertiary period. Primary porosity is intergranular and intragranular, and secondary processes are characterized by dissolution, intercrystalline, fracture and fenestral features. Diagenesis, through solution leaching and dolomitization, contributed greatly to porosity development. Reservoir traps of the Beda Formation are characterized by normal fault blocks and the general reservoir characteristics/properties appear to be facies controlled.
Resumo:
I will argue that the doctrine of eternal recurrence of the same no better interprets cosmology than pink elephants interpret zoology. I will also argue that the eternal-reiurn-of-the-same doctrine as what Magnus calls "existential imperative" is without possibility of application and thus futile. To facilitate those arguments, the validity of the doctrine of the eternal recurrence of the same will be tested under distinct rubrics. Although each rubric will stand alone, one per chapter, as an evaluation of some specific aspect of eternal recurrence, the rubric sequence has been selected to accommodate the identification of what I shall be calling logic abridgments. The conclusions to be extracted from each rubric are grouped under the heading CONCLUSION and appear immediately following rubric ten. Then, or if, at the end of a rubric a reader is inclined to wonder which rubric or topic is next, and why, the answer can be found at the top of the following page. The question is usually answered in the very first sentence, but always answered in the first paragraph. The first rubric has been placed in order by chronological entitlement in that it deals with the evolution of the idea of eternal recurrence from the time of the ancient Greeks to Nietzsche's August, 1881 inspiration. This much-recommended technique is also known as starting at the beginning. Rubric 1 also deals with 20th. Century philosophers' assessments of the relationship between Nietzsche and ancient Greek thought. The only experience of E-R, Zarathustra's mountain vision, is second only because it sets the scene alluded to in following rubrics. The third rubric explores .ii?.ih T jc,i -I'w Nietzsche's evaluation of rationality so that his thought processes will be understood appropriately. The actual mechanism of E-R is tested in rubric four...The scientific proof Nietzsche assembled in support of E-R is assessed by contemporary philosophers in rubric five. E-R's function as an ethical imperative is debated in rubrics six and seven.. .The extent to which E-R fulfills its purpose in overcoming nihilism is measured against the comfort assured by major world religions in rubric eight. Whether E-R also serves as a redemption for revenge is questioned in rubric nine. Rubric ten assures that E-R refers to return of the identically same and not merely the similar. In addition to assemblage and evaluation of all ten rubrics, at the end of each rubric a brief recapitulation of its principal points concludes the chapter. In this essay I will assess the theoretical conditions under which the doctrine cannot be applicable and will show what contradictions and inconsistencies follow if the doctrine is taken to be operable. Harold Alderman in his book Nietzsche's Gift wrote, the "doctrine of eternal recurrence gives us a problem not in Platonic cosmology, but in Socratic selfreflection." ^ I will illustrate that the recurrence doctrine's cosmogony is unworkable and that if it were workable, it would negate self-reflection on the grounds that selfreflection cannot find its cause in eternal recurrence of the same. Thus, when the cosmology is shown to be impossible, any expected ensuing results or benefits will be rendered also impossible. The so-called "heaviest burden" will be exposed as complex, engrossing "what if speculations deserving no linkings to reality. To identify ^Alderman p. 84 abridgments of logic, contradictions and inconsistencies in Nietzsche's doctrine of eternal recurrence of the same, I. will examine the subject under the following schedule. In Chapter 1 the ancient origins of recurrence theories will be introduced. ..This chapter is intended to establish the boundaries within which the subsequent chapters, except Chapter 10, will be confined. Chapter 2, Zarathustra's vision of E-R, assesses the sections of Thus Spoke Zarathustra in which the phenomenon of recurrence of the same is reported. ..Nihilism as a psychological difficulty is introduced in this rubric, but that subject will be studied in detail in Chapter 8. In Chapter 2 the symbols of eternal recurrence of the same will be considered. Whether the recurrence image should be of a closed ring or as a coil will be of significance in many sections of my essay. I will argue that neither symbolic configuration can accommodate Nietzsche's supposed intention. Chapter 3 defends the description of E-R given by Zarathustra. Chapter 4, the cosmological mechanics of E-R, speculates on the seriousness with which Nietzsche might have intended the doctrine of eternal recurrence to be taken. My essay reports, and then assesses, the argument of those who suppose the doctrine to have been merely exploratory musings by Nietzsche on cosmological hypotheses...The cosmogony of E-R is examined. In Chapter 5, cosmological proofs tested, the proofs for Nietzsche's doctrine of return of the same are evaluated. This chapter features the position taken by Martin ' Heidegger. My essay suggests that while Heidegger's argument that recurrence of the same is a genuine cosmic agenda is admirable, it is not at all persuasive. Chapter 6, E-R is an ethical imperative, is in essence the reporting of a debate between two scholars regarding the possibility of an imperative in the doctrine of recurrence. Their debate polarizes the arguments I intend to develop. Chapter 7, does E-R of the same preclude alteration of attitudes, is a continuation of the debate presented in Chapter 6 with the focus shifted to the psychological from the cosmological aspects of eternal recurrence of the same. Chapter 8, Can E-R Overcome Nihilism?, is divided into two parts. In the first, nihilism as it applies to Nietzsche's theory is discussed. ..In part 2, the broader consequences, sources and definitions of nihilism are outlined. My essay argues that Nietzsche's doctrine is more nihilistic than are the world's major religions. Chapter 9, Is E-R a redemption for revenge?, examines the suggestion extracted from Thus Spoke Zarathustra that the doctrine of eternal recurrence is intended, among other purposes, as a redemption for mankind from the destructiveness of revenge. Chapter 10, E-R of the similar refuted, analyses a position that an element of chance can influence the doctrine of recurrence. This view appears to allow, not for recurrence of the same, but recurrence of the similar. A summary will recount briefly the various significant logic abridgments, contradictions, and inconsistencies associated with Nietzsche's doctrine of eternal recurrence of the same. In the 'conclusion' section of my essay my own opinions and observations will be assembled from the body of the essay.
Resumo:
The Rankin Inlet area, on the west shore of Hudson Bay in the Northwest Territories, is in the Churchill Structural Province. Metamorphosed volcanic and sedimentary rocks, previously mapped as Archean and part of the Kaminak Group, underlie most of the area. The Rankin Inlet Group consists of greywacke, with minor conglomeratic greywacke, quartzite and dolomite, overlain by massive and pillowed basaltic flows. Gabbro sills intrude the sediments near the base of the volcanic sequence and three serpentinite sills outcrop at the base of the volcanic sequence. The sediments are in fault-contact with quartz monzonite to the south and were intruded by granitic rocks to the northwest. Two periods of folding were defined by the mapping. The first generation folds are recumbent isoclinal folds, with northwest-trending and northeast-dipping axial planes, formed through gravitational sliding. The second generation folds are symmetrically disposed about the axis of the granitic intrusion and have east-southeast trending and nearly vertical axial planes. Whole-rock analysis of 64 rock samples indicates that metasomatic alteration accompanied the intrusion of both the granitic rocks and the serpentinite. The volcanic rocks, gabbro and serpentinite were derived from a magma of oceanic tholeiitic affinities. The stratigraphic sequence and chemistry of the volcanic rocks of the Rankin Inlet Group indicate that this assemblage is correlative with the Hurwitz Group rather than the Kaminak Group and is therefore Aphebian in age.
Resumo:
The Sand Creek Prospect is located within the eastern exposed margin of the Coast Plutonic Complex. The occurrence is a plug and dyke porphyry molybdenum deposit. The rock types, listed in decreasing age: 1) metamorphlc schists and gneisses; 2) diorite suite rocks - diorite, quartz diorite, tonalite; 3) rocks of andesitic composition; 4) granodiorites, coarse porphyritic granodiorite, quartzfeldspar porphyry, feldspar porphyry; and 5) lamprophyre. Hydrothermal alteration is known to have resulted from emplacement of the hornblende-feldspar porphyry through to the quartz-feldspar porphyry. Molybdenum mineralization is chiefly associated with the quartz-feldspar porphyry. Ore mineralogy is dominated by pyrite with subordinate molybdenite, chalcopyrite, covelline, sphalerite, galena, scheelite, cassiterite and wolframite. Molybdenite exhibits a textural gradation outward from the quartz-feldspar porphyry. That is, disseminated rosettes and rosettes in quartz veins to fine-grained molybdenite in quartz veins and potassic altered fractures to fine-grained molybdenite paint or 6mears in the peripheral zones. The quartz-feldspar porphyry dykes were emplaced in an inhomogeneous stress field. The trend of dykes, faults and shear zones is 0^1° to 063° and dips between 58° NW and 86* SE. Joint Pole distribution reflects this fault orientation. These late deformatior maxima are probably superimposed upon annuli representing diapiric emplacement of the plutons. A model of emplacement involving two magmatic pulses is given in the following sequence: Diorite pulse (i) dioritequartz diorite, (ii) tonalites; granodiorite pulse (iii) hornblende-fildspar microporphyry, hornblende/biotite porphyry, (iv) coarse grained granodiorite, (v) quartz-feldspar porphyry, (vi) feldspar porphyry, and (vii) lamprophyre. The combination of plutonic and coarse porphyritic textures, extensive propylitic overprinting of potassic alteration assemblages suggests that the. prospect represents the lower reaches of a porphyry system.
Resumo:
The Pater metavolcanic suite (PVS) was extruded as part O'f the basal Pater Formation of the Huronian Supergroup ca. 2.4 Ga. They Ars classified as wi thin-plate tholeiites associated with an immature ri-fting episode, and are inter layered with associated vol cani clastic and metasedimentary units. Post-solidif ication alteration caused redistribution o-f the alkalies, Sr, Rb, Ba, Cu, and SiO^. Ce, Y, Zr, CFezOs (as total Fe), Al^Os, TiOa, and, PaOa are considered to have remained essentially immobile in least altered samples. Petrogenetic modelling indicates the PVS was derived from the partial melting of two geochemical ly similar sources in the sub-continental lithosphere. Fractionation was characterized by an oli vine-plagioclase assemblage and a sub-volcanic plagioclase-clinopyroxene assemblage. A comparative study indicates that enrichment of the postulated Huronian source cannot be reconciled by Archean contamination. Enrichment is thought to have been caused by hydrous veined metasomatic heterogeneities in the sub-continental lithosphere, generated by an Archean subduct ion event before 2.68 Ga.
Resumo:
A comprehensive elemental, isotopic and microstructural analyses was undertaken of brachiopod calcites from the Hamilton Group (Middle Devonian), Clinton Group (Middle Silurian) and Middle to Upper Ordovician strata of Ontario and New York State. The majority of specimens were microstructurally and chemically preserved in a pristine state, although a number of specimens show some degree of post-depositional alteration. Brachiopod calcites from the Hamilton and Clinton Groups were altered by marine derived waters whereas Trenton Group (Middle Ordovician) brachiopods altered in meteorically derived fluids. Analysis of the elemental and isotopic compositions of pristine Hamilton Group brachiopods indicates there are several chemical relationships inherent to brachiopod calcite. Taxonomic differentiation of Mg, Sr and Na contents was evident in three co-occuring species from the Hamilton Group. Mean Mg contents of pristine brachiopods were respectively Athyris spiriferoides (1309ppm), Mucrospirifer mucronatus (1035ppm) and Mediospirifer audacula (789ppm). Similarly, taxonomic differentiation of shell calcite compositions was observed in co-occuring brachiopods from the Clinton Group (Middle Silurian) and the Trenton Group (Middle Ordovician). The taxonomic control of elemental regulation into shell calcite is probably related to the slightly different physiological systems and secretory mechanisms. A relationship was observed in Hamilton Group species between the depth of respective brachiopod communities and their Mg, Sr and Na contents. These elements were depleted in the shell calcites of deeper brachiopods compared to their counterparts in shallower reaches. Apparently shell calcite elemental composition is related to environmental conditions of the depositional setting, which may have controlled the secretory regime, mineral morphology of shell calcite and precipitation rates of each species. Despite the change in Mg, Sr and Na contents between beds and formations in response to environmental conditions, the taxonomic differentiation of shell calcite composition is maintained. Thus, it may be possible to predict relative depth changes in paleoenvironmental reconstructions using brachiopod calcite. This relationship of brachiopod chemistry to depth was also tested within a transgressiveregressive (T-R) cycle in the Rochester Shale Formation (Middle Silurian). Decreasing Mg, Sr and Na contents were observed in the transition from the shallow carbonates of the Irondequoit Formation to the deeper shales of the lowest 2 m of Rochester Shale. However, no isotopic and elemental trends were observed within the entire T-R cycle which suggests that either the water conditions did not change significantly or that the cycle is illusory. A similar relationship was observed between the Fe and Mn chemistries of shell calcite and redox/paleo-oxygen conditions. Hamilton Group brachiopods analysed from deeper areas of the shelf are enriched in Mn and Fe relative to those from shallow zones. The presence of black shales and dysaerobic faunas, during deposition of the Hamilton Group, suggests that the waters of the northern Appalachian Basin were stratified. The deeper brachiopods were marginally positioned above an oxycline and their shell calcites reflect periodic incursions of oxygen depleted water. Furthermore, analysis of Dalmanella from the black shales of the Collingwood Shale (Upper Ordovician) in comparison to those from the carbonates of the Verulam Formation (Middle Ordovician) confirm the relationship of Fe and Mn contents to periodic but not permanent incursions of low oxygen waters. The isotopic compositions of brachiopod calcite found in Hamilton Group (813C; +2.5% 0 to +5.5% 0; 8180 -2.50/00 to -4.00/00) and Clinton Group (813C; +4.00/00 to +6.0; 8180; -1.8% 0 to -3.60/ 00) are heavier than previously reported. Uncorrected paleotemperatures (assuming normal salinity, 0% 0 SMOW and no fractionation effects) derived from these isotopic values suggest that the Clinton sea temperature (Middle Silurian) ranged from 18°C to 28°C and Hamilton seas (Middle Devonian) ranged between 24°C and 29°C. In addition, the isotopic variation of brachiopod shell calcite is significant and is related to environmental conditions. Within a single time-correlative shell bed (the Demissa Bed; Hamilton Group) a positive isotopic shift of 2-2.5% 0 in 013C compositions and a positive shift of 1.0-1.50/00 in 0180 composition of shell calcite is observed, corresponding with a deepening of brachiopod habitats toward the axis of the Appalachian Basin. Moroever, a faunal succession from deeper Ambocoelia dominated brachiopod association to a shallow Tropidoleptus dominated assocation is reflected by isotopic shifts of 1.0-1.50/00. Although, other studies have emphasized the significance of ±20/oo shifts in brachiopod isotopic compositions, the recognition of isotopic variability in brachiopod calcite within single beds and within depositional settings such as the Appalachian Basin has important implications for the interpretation of secular isotopic trends. A significant proportion of the variation observed isotopic distribution during the Paleozoic is related to environmental conditions within the depositional setting.
Resumo:
Owing to the fact that low-Mg calcite fossil shells are so important in paleoceanographic research, 249 brachiopod, cement and matrix specimens from two neighboring localities (Jemez Springs and Battleship Rock), of the Upper Pennsylvanian Madera Formation were analyzed. Of which, about 86% of the Madera brachiopods are preserved in their pristine mineralogy, microstructure and geochemistry. Cement and matrix samples, in contrast, have been subjected to complete but variable post-deposition~1 alteration. It is confirmed that the stable isotope data of brachiopods are much better than that of matrix material in defining depositional parameters. Because there is no uniform or constant relationship between the two data bases (e.g., from 0.1 to 3.0%0 for 0180 and from 0.2 to 6.7%0 for 013C in this study), it is not possible to make corrections for the matrix data. Regarding the two stratigraphic sections, elemental and petrographic analyses suggest that Jemez Springs is closer to Penasco Uplift than Battleship Rock. Seawater at Jemez Springs is more aerobic, and the water chemistry is more influenced by continental sources than that at Battleship Rock. In addition, there is a relatively stronger dolomitization in the mid-section of the Battleship Rock. Results further suggest that no significant biogenic fractionation or vital effects occurred during their shell secretion, suggesting that the Madera brachiopods incorporated oxygen and carbon isotopes in equilibrium with the ambient seawater. This conclusion is not only drawn from the temporal and spatial analyses, but also supported by brachiopod inter-generic comparison (Composita and Neospirifer) and statistical analysis ( t-test).
Resumo:
Intracerebroventricular (ICV) administration of bombesin (BN) induces a
syndrome characterized by stereotypic locomotion and grooming,
hyperactivity and sleep elimination, hyperglycemia and hypothermia,
hyperhemodynamics, feeding inhibition, and gastrointestinal function
changes. Mammalian BN-like peptides (MBNs), e.g. gastrin-releasing
peptide (GRP), Neuromedin C (NMC), and Neuromedin B (NMB), have been
detected in the central nervous system. Radio-labeled BN binds to specific
sites in discrete cerebral regions. Two specific BN receptor subtypes (GRP
receptor and NMB receptor) have been identified in numerous brain regions.
The quantitative 2-[14C]deoxyglucose ([14C]20G) autoradiographic
method was used to map local cerebral glucose utilization (LCGU) in the
rat brain following ICV injection of BN (vehicle, BN O.1Jlg, O.5Jlg). At each
dose, experiments were conducted in freely moving or restrained
conditions to determine whether alterations in cerebral function were the
result of BN central administration, or were the result of BN-induced
motor stereotypy. The anteroventral thalamic nucleus (AV) (p=O.029),
especially its ventrolateral portion (AVVL) (p
Resumo:
Recombinant human adenovirus (Ad) vectors are being extensively explored for their use in gene therapy and recombinant vaccines. Ad vectors are attractive for many reasons, including the fact that (1) they are relatively safe, based on their use as live oral vaccines, (2) they can accept large transgene inserts, (3) they can infect dividing and postmitotic cells, and (4) they can be produced to high titers. However, there are also a number of major problems associated with Ad vectors, including transient foreign gene expression due to host cellular immune responses, problems with humoral immunity, and the creation of replication competent adenoviruses (RCA). Most Ad vectors contain deletions in the E1 region that allow for insertion of a transgene. However, the E1 gene products are required for replication and thus must be supplied in trans by a helper ceillille that will allow for the growth and packaging of the defective virus. For this purpose the 293 cell line (Graham et al., 1977) is used most often; however, homologous recombination between the vector and the cell line often results in the generation of RCA. The presence of RCA in batches of adenoviral vectors for clinical use is a safety risk because tlley . may result in the mobilization and spread of the replication-defective vector viruses, and in significant tissue damage and pathogenicity. The present research focused on the alteration of the 293 cell line such that RCA formation can be eliminated. The strategy to modify the 293 cells involved the removal of the first 380 bp of the adenovirus genome through the process of homologous recombination. The first step towards this goal involved identifying and cloning the left-end cellular-viral jUl1ction from 293 cells to assemble sequences required for homologous recombination. Polymerase chain reaction (PCR) was performed to clone the junction, and the clone was verified through sequencing. The plasn1id PAM2 was then constructed, which served as the targeting cassette used to modify the 293 cells. The cassette consisted of (1) the cellular-viral junction as the left-end region of homology, (2) the neo gene to use for positive selection upon tranfection into 293 cells, (3) the adenoviral genome from bp 380 to bp 3438 as the right-end region of homology, and (4) the HSV-tk gene to use for negative selection. The plasmid PAM2 was linearized to produce a double strand break outside the region of homology, and transfected into 293 cells using the calcium-phosphate technique. Cells were first selected for their resistance to the drug G418, and subsequently for their resistance to the drug Gancyclovir (GANC). From 17 transfections, 100 pools of G418f and GANCf cells were picked using cloning lings and expanded for screening. Genomic DNA was isolated from the pools and screened for the presence of the 380 bps using PCR. Ten of the most promising pools were diluted to single cells and expanded in order to isolate homogeneous cell lines. From these, an additional 100 G41Sf and GANef foci were screened. These preliminary screening results appear promising for the detection of the desired cell line. Future work would include further cloning and purification of the promising cell lines that have potentially undergone homologous recombination, in order to isolate a homogeneous cell line of interest.
Resumo:
The neuropeptide Th1RFamide with the sequence Phe-Met-Arg-Phe-amide was originally isolated in the clam Macrocallista nimbosa (price and Greenberg, 1977). Since its discovery, a large family ofFl\1RFamide-related peptides termed FaRPs have been found to be present in all major animal phyla with functions ranging from modulation of neuronal activity to alteration of muscular contractions. However, little is known about the genetics encoding these peptides, especially in invertebrates. As FaRP-encoding genes have yet to be investigated in the invertebrate Malacostracean subphylum, the isolation and characterization ofFaRP-encoding DNA and mRNA was pursued in this project. The immediate aims of this thesis were: (1) to amplify mRNA sequences of Procambarus clarkii using a degenerate oligonucleotide primer deduced from the common amino acid sequence ofisolated Procambarus FaRPS, (2) to determine if these amplification products encode FaRP gene sequences, and (3) to create a selective cDNA library of sequences recognized by the degenerate oligonucleotide primer. The polymerase chain reaction - rapid amplification of cDNA ends (PCR-RACE) is a procedure in which a single gene-specific primer is used in conjunction with a generalized 3' or 5' primer to amplify copies ofthe region between a single point in the transcript and the 3' or 5' end of cDNA of interest (Frohman et aI., 1988). PCRRACE reactions were optimized with respect to primers used, buffer composition, cycle number, nature ofgenetic substrate to be amplified, annealing, extension and denaturation temperatures and times, and use of reamplification procedures. Amplification products were cloned into plasmid vectors and recombinant products were isolated, as were the recombinant plaques formed in the selective cDNA library. Labeled amplification products were hybridized to recombinant bacteriophage to determine ligated amplification product presence. When sequenced, the five isolated PCR-RACE amplification products were determined not to possess FaRP-encoding sequences. The 200bp, 450bp, and 1500bp sequences showed homology to the Caenorhabditis elegans cosmid K09A11, which encodes for cytochrome P450; transfer-RNA; transposase; and tRNA-Tyr, while the 500bp and 750bp sequences showed homology with the complete genome of the Vaccinia virus. Under the employed amplification conditions the degenerate oligonucleotide primer was observed to bind to and to amplify sequences with either 9 or 10bp of 17bp identity. The selective cDNA library was obselVed to be of extremely low titre. When library titre was increased, white. plaques were isolated. Amplification analysis of eight isolated Agt11 sequences from these plaques indicated an absence of an insertion sequence. The degenerate 17 base oligonucleotide primer synthesized from the common amino acid sequence ofisolated Procambarus FaRPs was thus determined to be non-specific in its binding under the conditions required for its use, and to be insufficient for the isolation and identification ofFaRP-encoding sequences. A more specific primer oflonger sequence, lower degeneracy, and higher melting temperature (TJ is recommended for further investigation into the FaRP-encoding genes of Procambarlls clarkii.
Resumo:
Trilobites ¥tere collected from Ordovician and Devonian formations of Ontario} New York} Ohio} Oklahoma} and Indiana. Diversity was generally low} but 19..?telllS and Ph..~tY>ps ¥tere the most abundant species from the Ordovician and Devonian} respectively. Recent marine arthropods ¥tere collected from the Atlantic shore of the middle Florida Keys} and from the Pacific and lagoonal waters at Cape Beale} B. C. Fresh-water arthropods were collected along the shore of the Severn River in northcentral Ontario. Cuticles ¥tere analyzed for major} minor and trace elements, 180 and 13C isotopes, as ¥tell as examined by scanning electron micr?scope to identify original and diagenetic fabrics. Examination of trilobite cuticles by scanning electron microscope revealed several microstructures consistent with those observed in Recent arthropods. Microstructures} such as setae and tegumental gland duct openings} in like sized Lim/IllS and Isoteline trilobites may indicate common ancestral origins for these organisms, or simply parallel cuticle evolutions. The dendritic microstructure, originally' thought to be a diagenetic indicator, was found in Recent specimens and therefore its presence in trilobites may be suggestive of the delicate nature of diagenesis in trilobites. The absence of other primary microstructures in trilobites may indicate alteration, taxonomic control} or that there is some inherent feature of S EM examination which may' not allow detection of some features} while others are apparently visit·le onl~1 under SH.·1. The region of the cuticle sampled for examination is also a major influence in detecting pristine microstructures, as not all areas of trilobite and Recent arthropod cuticles will have microstructures identifiable in a SEM study. Subtleties in the process of alteration, however} ma~·· leave pristine microstructures in cuticles that are partial~/ silicified or do 10m itized, and degree and type of alteration may vary stratigraphically and longitudinally within a unit. The presence of fused matrices, angular calcite rhombs, and pyrite in the cuticle are thought to be indicative of altered cuticles, although pyritization may not affect the entire cuticle. t-~atural processes in Recent arthropods, such as molting, lead to variations in cuticle chemistries, and are thought to reflect the area of concentration of the elements during calcification. The level of sodium in Recent arthropods was found to be higher than that in trilobites, but highly mobile when sUbjected to the actions of VY'€'athering. Less saline water produced lovy'€'r magnesium and higher calcium values in Recent specimens .. and metal variations in pristine Ordovician trilobite cuticle appears to follow the constraints outlined for Recent arthropods, of regulation due to the chemislry of the surrounding medium. In diagenetic analysis, sodium, strontium and magnesium proved most beneficial in separating altered from least altered trilobites. Using this criterion, specimens from shale show the least amount of geochemical alteration, and have an original mineralogy of 1.7 - 2.4 mole % MgC03 (8000 t(> 9500 ppm magnesium) for both /s>..?/e/11S lJA'i.riff!11S and PseIAit'11J17ites I..itmirpin..itl/~ and 2.8 - 3.3 mole % MgC03 (5000 to 7000 ppm magnesium) for Ph.i{).?PS This is Slightly lower than the mineralogy of Recent marine arthropods (4.43 - 12.1 mole % MgC03), and slightly higher than that of fresh-water crayfish (0.96 - 1.82 mole % MgC03). Geochemically pristine trilobites were also found to possess primary microstructures. Stable isotope values and trends support the assertion that marine-meteoriclburial fluids were responsible for the alteration observed in a number of the trilobite specimens. The results of this stUdy suggest that fossil material has to be evaluated separately along taxonomic and lithological lines to arrive at sensible diagenetic and e nvironmenta I interpretations.
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Geochemical examination of the rock matrix and cements from core material extracted from four oil wells within southwestern Ontario suggest various stages of diagenetic alteration and preservation of the Trenton Group carbonates. The geochemical compositions of Middle Ordovician (LMC) brachiopods reflect the physicochemical water conditions of the ambient depositional environment. The sediments appear to have been altered in the presence of mixed waters during burial in a relatively open diagenetic microenvironment. Conodont CAl determination suggests that the maturation levels of the Trenton Group carbonates are low and proceeded at temperatures of about 30 - 50°C within the shallow burial environment. The Trenton Group carbonates are characterized by two distinct stages of dolomitization which proceeded at elevated temperatures. Preexisting fracture patterns, and block faulting controlled the initial dolomitization of the precursor carbonate matrix. Dolomitization progressed In the presence of warm fluids (60 75°C) with physicochemical conditions characteristic of a progressively depleted basinal water. The matrix is mostly Idiotopic-S and Idiotopic-E dolomite, with Xenotopic-A dolomite dominating the matrix where fractures occur. The second stage of dolomitization involved hydrothermal basinal fluid(s) with temperatures of about 60 - 70°C. These are the postulated source for the saddle dolomite and blocky calcite cements occurring in pore space and fractures. Rock porosity was partly occluded by Idiotopic-E type dolomite. Late stage saddle dolomite, calcite, anhydrite, pyrite, marcasite and minor sphalerite and celestite cements effectively fill any remaining porosity within specific horizons. Based on cathode luminescence, precipitation of the different diagenetic phases probably proceeded in open diagenetic systems from chemically homogeneous fluids. Ultraviolet fluorescence of 11 the matrix and cements demonstrated that hydrocarbons were present during the earliest formation of saddle dolomite. Oxygen isotope values of -7.6 to -8.5 %0 (PDB), and carbon isotope values of - 0.5 and -3.0 %0 (PDB) from the latest stage dog-tooth calcite cement suggest that meteoric water was introduced into the system during their formation. This is estimated to have occurred at temperatures of about 25 - 40°C. Specific facies associations within the Trenton Group carbonates exhibit good hydrocarbon generating potential based on organic carbon preservation (1-3.5%). Thermal maturation and Lopatin burial-history evaluations suggest that hydrocarbons were generated within the Trenton Group carbonates some time after 300 Ma . Progressively depleted vanadium trends measured from hydrocarbon samples within southwestern Ontario suggests its potential use as a hydrocarbon migration indicator on local (within an oilfield) and on regional scales.