Dissertations / Theses on the topic 'Biochemical Mice'

Thesis (Ph.D.)--University of Melbourne, The Walter & Eliza Hall Institute of Medical research, Dept. of Medical Biology, 2004.<br>Typescript (photocopy). Includes bibliographical references (leaves 161-190).

Recent advances in molecular genetics have enabled generation of sophisticated genetically-modified mouse models to study specific molecules and their biological function in vivo. Here, I investigated biochemical changes in two different genetically-modified mouse lines with previously described learning and memory phenotypes. Firstly, I analysed biochemical changes in a mouse line carrying a threonine to alanine point mutation at Thr286 of alpha Ca2+/calmoduline-dependent kinase II (aCaMKII), which disenables this phosphorylation site. Autophosphorylation at Thr286 switches aCaMKII into an autonomous activity mode. The T286A mutant mice displayed changes in basal phosphorylation levels. In order to study biochemical changes after activity-dependent synaptic potentiation, an in vivo long-term potentiation (LTP) approach was established and validated by assessing activity-dependent changes in phosphorylation levels of well-characterised marker molecules including synapsin I and NR2B. Both aCaMKII and pCaMKII exhibited elevated levels of autophosphorylation after LTP stimulation in hippocampal area CA1 and dentate gyrus (DG). This finding indicates that pCaMKII may compensate for the loss of autonomous aCaMKII activity in T286A mutants. Furthermore, induction of LTP triggered phosphorylation of glycogen synthase kinase 3 (GSK3) at its inhibitory site suggesting a role for GSK3 in synaptic plasticity. Secondly, I investigated a transgenic (TG) mouse line expressing the cyclin-dependent kinase (Cdk5) activator protein, p25, a protein previously linked to some aspects of Alzheimer's disease (AD). The forebrain restricted expression of p25 started postnatally and stayed constant throughout the life-span of the TG mice. The expression of p25 triggered constitutive over-activation of Cdk5 in the TG mice. The p25 TG mice displayed age-dependent hyperphosphorylation of the microtubule-associated protein tail and age-dependent alterations in the processing of amyloid precursor protein (APP). Furthermore, p25-induced over-activation of Cdk5 led to inhibition of GSK3. This negative regulation of GSK3 was lost in aged p25 TG mice and correlated with the increased tau hyperphosphorylation. The levels of tau phosphorylation in aged p25 mice were reduced after treatment with lithium, an inhibitor of GSK3. These results indicate that GSK3 directly mediates tau hyperphosphorylation, whereas Cdk5 acts indirectly via inhibitory control of GSK3.

Nitrous oxide (N2O) oxidizes vitamin B12. disrupting deoxyribonucleic acid (DNA) synthesis. Occupational exposures to subanesthetic levels of the gas have been documented that may result in suppressed proliferative cell activities. Male CD-I mice were exposed to 0, 50, 500, and 5000 parts of N2O per million parts of air (ppm) for 6 hr/day, 5 days/week for 2 and 13 weeks. Splenic lymphocytes were assayed for responsiveness to mitogens and for the ability to produce interleukin-2 (lL-2) . Tritiated-thymidine ([3H]-TdR) uptake was measured in CD-I splenic lymphocytes cultured in a mixed-lymphocyte culture (MLC). Cytolytic cell activity was measured by 51chromium release assay. Antibody-mediated immunocompetency was determined for sheep red blood cell (SRBC)-sensitized animals by plaque-forming cell (PFC) assay and sera anti-SRBC antibody titer. Deoxyuridine suppression tests (dUdRST) were performed on bone marrow cells. Serum adrenocorticotropic hormone and corticosterone levels were determined. There was significantly decreased splenic lymphocyte uptake of [3H)-TdR by cells cultured with mitogenic substances and in MLC following 2-week animal exposures to 5000 ppm. After 13-week exposures, the animals' splenic lymphocytes showed decreased [3H]-TdR uptake following low N20 dosing and nonsignificantly increased responsiveness at the higher gas exposures in both the blastogenic and MLC assays. Compared to control animals, the 5000- ppm-exposure group had significantly depressed PFC activity and circulating anti-SRBC immunoglobulin M levels following 13-week gas exposures, and all three subchronic exposure groups demonstrated both decreased liver weights and leukopenia. Bone marrow activity at these dosing levels was dose-responsively depressed following subchronic gas exposures.No hormonal effect appears to be attributable to N20 exposure.

Includes bibliographical references (leaves 142-168) Aim was the purification and identification of the early primitive ectoderm-like (EPL) cell induction signals within the medium conditioned by the human hepatocellular carcinoma cell line HepG2 and the localisation of the signals that induce EPL cell and primitive ectoderm formation.

Hyperhomocysteinemia is a risk factor for cardiovascular disease and stroke. Nutritional and/or genetic disruptions in homocysteine metabolism can cause hyperhomocysteinemia. Mild methylenetetrahydrofolate reductase (MTHFR) deficiency due to the 677C &rarr; T mutation in the MTHFR gene is the most common genetic cause of hyperhomocysteinemia. The 677C &rarr; T variant is associated with an increased risk for neural tube defects, pregnancy complications, schizophrenia and Down syndrome, and with a decreased risk for colon cancer and leukemia. This variant is also a potential risk factor for vascular disease. Severe MTHFR deficiency results in homocystinuria, an inborn error of metabolism with neurological and vascular complications. We have generated mice with a knockout of the Mthfr gene. The Mthfr-deficient mice exhibit hyperhomocysteinemia and decreased methylation capacity. The Mthfr+/- mice appear normal, whereas the Mthfr-/- mice are smaller and have reduced survival. Abnormal external granule neuron development associated with increased cell death in the cerebellum was observed in the Mthfr-/- mice.<br>Evidence for cardiovascular pathology was obtained in several ways. Impaired aortic relaxation response to acetylcholine was seen in the Mthfr +/- mice fed a high methionine diet. Both Mthfr+/- and Mthfr-/- mice fed a low folate high methionine diet developed myocardial fibrosis in the left ventricle. Abnormal lipid deposition in the proximal portion of the aorta was observed in older Mthfr+/- and Mthfr-/- mice. After crossing Mthfr -deficient mice with apoE-null mice, we demonstrated that MTHFR deficiency promoted atherogenesis and its progression in the apoE-null mice.<br>Gene expression in brain of Mthfr-deficient mice was investigated via microarray analysis. Five genes with altered expression in the brain of Mthfr-/- mouse were validated by RT-PCR. In biochemical studies of human MTHFR, both FAD and folate were shown to stabilize the purified recombinant wild type and mutant MTHFRs from the baculovirus expression system against heat inactivation. The effect of folate appeared to be secondary to that of FAD, and S-adenosylmethionine (SAM) inhibited purified wild type and mutant MTHFRs with similar efficiency.<br>This dissertation will significantly contribute to our understanding of the role of MTHFR in human disease.

It is submitted in this thesis that the degree of activation or inhibition of macrophage function may differ in N. asteroides and N. brasiliensis infections with respect to release of plasminogen activator and of lysozyme The pattern of secretion of plasminogen activator and lysozyme in N. asteroides infections appears to differ in N. brasiliensis infection; and there is possibly a difference in the amount of lysozyme released by 2 day N. asteroides-activated macrophages and 2 day N. brasiliensis -activated macrophages. Strains of Nocardia organism did not influence macrophage morphology or ultrastructure. The study also shows the biochemical characteristics of plasminogen activator and lysozyme release, but not macrophage morphology and ultrastructure, are modified in the first 21 days of experimental Nocardia infections. There are three apparent mechanisms by which virulent strains of N. asteroides manage to survive within macrophages: (i) an ability to inhibit phagosome-lysozome fusion: (ii) alteration in the intraphagosomal pH: and (iii) alteration in the activity of the lysozomal enzyme acid-phosphatase. This study attempted to elucidate further the mechanisms enabling Nocardia organisms to persist and grow within macrophages. Reduced lysozyme release reflects diminished functional status of the macrophages of mice inoculated with N. asteroides or N. brasiliensis at certain times during infection. Reduced intracellular lysozyme levels have been linked with defects in bactericidal function. Such a reduction in intracellular and consequently extracellular levels of lysozyme might explain the capacity of Nocardia to survive intracellularly and to proliferate in the macrophage host.

La mucopolisacaridosi tipus VII (MPS VII) és una malaltia monogènica molt rara inclosa en el grup de malalties lisosòmiques. Està causada per la manca d’activitat β-­‐ glucuronidasa, un enzim lisosòmic involucrat en la via de degradació de glicosaminoglicans. Aquesta alteració congènita causa una disfunció del sistema lisosòmic que provoca una acumulació anormal als lisosomes i la disrupció de l’homeòstasi de la cèl·lula. La malaltia presenta diferents graus de severitat clínica que van des de la mort prenatal fins a pacients amb formes lleus i una esperança de vida de 20 o 30 anys. Entre els casos que presenten un inici postnatal de la malaltia, la forma més severa de MPS VII es caracteritza per hepatomegàlia, esplenomegàlia, anomalies esquelètiques, desenvolupament tardà i retard mental, entre altres símptomes. Actualment els tractaments per als pacients de MPS VII són intervencions per pal·liar els símptomes de la malaltia, però no disposen de cap tractament curatiu. La teràpia gènica és una estratègia prometedora de cara a trobar una cura per a malalties monogèniques. Entre els vectors de teràpia gènica disponibles, els virus adeno-­‐associats (AAV) presenten certes característiques que els fan molt atractius per a la teràpia gènica: permeten la transducció tant de cèl·lules en divisió com quiescents, proporcionen una expressió del transgèn a llarg termini, no poden replicar-­‐se de manera autònoma sense un virus accessori i, a més, les infeccions naturals dels AAV no són patogèniques. Diferents serotips d’AAV s’han utilitzat com a vectors de teràpia gènica en estudis preclínics. Entre ells, els serotips AAV9 i AAVrh10 són els que presenten una capacitat de transducció més gran, així com una gamma més àmplia de tipus cel·lulars que poden transduir, especialment al sistema nerviós central. El serotip AAVrh10 no és d’origen humà. Per això, el fet d’utilitzar-­‐lo com a vector de teràpia gènica podria evitar la neutralització per part dels factors immunològics específics contra AAV, factors que són presents en el sèrum humà després d’infeccions d’AAV naturals. Tanmateix, el reconeixement creuat per part d’anticossos generats contra AAV2, el serotip humà més comú, podrien interferir en el resultat de la teràpia. En aquest treball es proposa una estratègia de teràpia gènica per a MPS VII basada en una única injecció intratecal d’un vector AAVrh10 que conté el gen de la β-­‐ glucuronidasa, testada en ratolins MPS VII adults joves. Es mostra que l’administració del vector al líquid cefaloraquidi (LCR) per punció lumbar, una tècnica molt poc invasiva, permet la transducció d’estructures del sistema nerviós central (SNC) utilitzant una dosi de vector més baixa que mitjançant injecció intravenosa. A més, el drenatge del vector del LCR cap al torrent sanguini comporta la transducció d’òrgans somàtics com el fetge. Això genera una font de β-­‐glucuronidasa que aconsegueix una activitat enzimàtica en sèrum comparable a la de ratolins sans. L’expressió sostinguda de l’enzim recombinant per part de les cèl·lules transduïdes, així com la correcció creuada deguda a la secreció de l’enzim, aconsegueixen la correcció de les característiques bioquímiques i histopatològiques de la malaltia, tant al SNC com als òrgans somàtics. Aquesta correcció a nivell cel·lular condueix a una millora significativa de les capacitats físiques, cognitives i emocionals dels ratolins MPS VII i aconsegueix doblar la seva esperança de vida.<br>Mucopolysaccharidosis type VII (MPS VII) is an ultrarare monogenic lysosomal storage disease. It is caused by the lack of β-­‐glucuronidase activity, a lysosomal enzyme involved in the degradation pathway of glycosaminoglycans. This inborn genetic alteration causes a dysfunction of the lysosomal system that entails abnormal lysosomal storage and disruption of cell homeostasis. The disease presents a range of clinical severity among patients, from death in utero to a life expectancy of up to 20 or 30 years for the milder forms. The severe form of MPS VII among cases with postnatal disease onset is characterized by hepatosplenomegaly, skeletal abnormalities, developmental delay and mental retardation, among other symptoms. Currently, the only treatments for MPS VII patients are interventions to alleviate the symptoms, but no curative treatment is available. Gene therapy is a promising therapeutic approach to find a cure for monogenic diseases. Among the available gene delivery vectors, adeno-­‐associated viruses (AAVs) present several features that make them attractive for gene therapy strategies: they are able to transduce dividing and quiescent cells, they provide long term expression of the transgene, they are not able to autonomously replicate without a helper virus, and wild type AAV infections are not pathogenic. Different AAV serotypes have been used as gene therapy vectors in preclinical studies. Among them, AAV9 and AAVrh10 are the serotypes which show greater transduction capacity and a broader range of cell-­‐type specific tropism, particularly in the central nervous system. The use of AAVrh10, a non-­‐human serotype, may avoid the neutralization by anti-­‐AAV immune factors present in human sera after natural AAV infections. However, cross-­‐reactivity with antibodies raised against AAV2, the most common human AAV serotype, may still interfere in the therapeutic outcome. In this work, we propose a gene therapy strategy for MPS VII based on a single intrathecal injection of an AAVrh10 coding for the β-­‐glucuronidase gene, tested in young adult MPS VII mice. We show that vector delivery to the CSF by lumbar puncture, a poorly invasive technique, allows the transduction of CNS structures using a lower vector dose than by intravenous delivery. In addition, the drainage of the vector from the CSF to the bloodstream results in transduction of somatic organs such as liver, thus providing a systemic β-­‐glucuronidase source that achieves serum enzymatic activity comparable to wild type mice. The sustained recombinant enzyme expression by AAV-­‐transduced cells, and the cross-­‐correction provided by enzyme secretion, attains the correction of biochemical and histopathological hallmarks of the disease in CNS and somatic organs. This correction at the cellular level leads to a significant improvement of physical, cognitive and emotional characteristics of MPS VII mice and a doubling of the MPS VII mouse life span.

Thesis (Ph. D.)--University of Missouri-Columbia, 2006.<br>Title from title screen of research.pdf file (viewed on December 22, 2006). The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. "May 2006" Includes bibliographical references.

Abstract Genomic clones covering the entire mouse type XIII collagen gene (Col13a1) were isolated, and the complete exon-intron organization was characterized. The gene was found to be about 135 kb in size and to locate in the mouse chromosome 10. Comparison of gene structures and promoter regions between man and mouse indicated high conservation between the two species. In order to understand the biological function of type XIII collagen, a mouse line that expresses type XIII collagen with replacement of the cytosolic and transmembrane domains by a short, non-descript sequence was generated using homologous recombination. Expression of this aminoterminally altered type XIII collagen led to mild but progressive muscular atrophy in mice. The integrity of muscle cells was disturbed and the basement membrane showed areas of detachment from the sarcolemma as well as clearly altered structure at myotendinous junctions. These phenotypical changes were, nevertheless, local, since the majority of the muscle was intact. The results show the importance of the membrane anchorage of the type XIII collagen protein in adhesion and, consequently in the maintenance of muscle integrity. To study the significance of various regions of type XIII collagen, wild-type and mutant forms of the protein were produced recombinantly in insect cells. The transmembrane domain and the adjacent region of ectodomain were found to be crucial for the formation of type XIII collagen molecules with all of the three collagenous domains in trimeric conformation. A previously characterized conserved membrane-proximal region of the ectodomain was predicted to harbour a coiled-coil conformation. This was suggested to begin in the transmembrane domain of type XIII collagen and in several other collagenous transmembrane proteins. Type XIII collagen lacking this coiled-coil sequence was correctly folded with respect to its central COL2 and carboxylterminal COL3 domains. Between them, in the NC3 domain, a second coiled-coil sequence was found, and this was suggested to function as a second association region. The second coiled-coil sequence was found to be conserved in the two other type XIII collagen-like molecules as well. To obtain precise information about the location and level of type XIII collagen expression, a reporter mouse line synthesizing a recombinant protein with the cytoplasmic and transmembrane portions of type XIII collagen linked in-frame with the β-galactosidase enzyme was generated. The reporter mice showed high expression of type XIII collagen at neuromuscular junctions and in the periosteum of bone. Interestingly, the growth of the reporter mice was reduced at puberty. Their long bones showed a decreased diameter and impaired mechanical properties. In addition, their peripheral nerves showed areas of detachment from muscle cells at neuromuscular junctions. These results provide further evidence for the role of type XIII collagen in cell adhesion. They also show the importance of proper adhesion conducted by type XIII collagen in signaling between the extracellular matrix and cells and in the cellular response.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-06-20T15:46:28Z No. of bitstreams: 1 2010 - Carla Rodrigues Bahiense.pdf: 694207 bytes, checksum: 012f3bf71990c09f742e165f567b3e77 (MD5)<br>Made available in DSpace on 2017-06-20T15:46:28Z (GMT). No. of bitstreams: 1 2010 - Carla Rodrigues Bahiense.pdf: 694207 bytes, checksum: 012f3bf71990c09f742e165f567b3e77 (MD5) Previous issue date: 2010-07-13<br>Belonging to the Order Psittaciformes, Family Psittacidae, the Ara ararauna, like other parrots, is a Brazilian bird much exploited in the pet market, which makes it a frequent veterinary patient. Despite of the extreme clinical relevance of laboratory examinations, data on hematology and serum biochemistry of this species are scarce. Several factors can interfere with the birds results of hematological and biochemical values, such as age, sex, reproductive period, among others. The study aimed to recognize patterns and to determine haematological and serum biochemical parameters for the species in question, explaining possible variations related to sex, management and method of restraint. In the experiment was used 68 specimens, 33 from a commercial breeding facility and 35 from the live collection of the RIOZOO Foundation. The blood samples were collected by jugular vein, transferred into tubes containing EDTA and other with no anticoagulant. Only the birds of RIOZOO were subjected to a new collection, which occurred 10 minutes after of anesthesia with isoflurane, thus forming a group of 33 animals anesthetized and another 35 unanesthetized. Of each animal were determined trombocyte conts, erythrocyte counts, total and specific leucocyte counts, concentrations of total plasmatic protein, urea, creatinine, uric acid, cholesterol, tryglicer?des, total serum protein, albumin and globulin, and activityes of alkaline phosphatase, aspartate aminotransferase, amylase, lipase, and creatine-kinase enzymes. The results were analyzed by ANOVA and paired T tests. Females had higher trombocitometria. Non-anesthetized animals had a greater total WBC count. Weren?t significant differences between anesthetized males and females. The birds originated from RIOZOO had higher values of PCV, red blood cell count, MCV, eosinophils, basophils, urea, AST and CK, however, lower values of albumin and creatinine. The study revealed that was a significant level of discrepancies between the different groups, allowing the creation of a standard more specific hematologic, according to the individual characteristics of each patient.<br>Pertencente ? Ordem Psittaciforme e Fam?lia Psittacidae, a Ara ararauna, assim como os demais psitac?deos, ? uma ave brasileira muito explorada no mercado pet, o que a torna um freq?ente paciente veterin?rio. Apesar da extrema relev?ncia cl?nica dos exames laboratoriais, dados acerca da hematologia e bioqu?mica s?rica dessa esp?cie ainda s?o escassos. Importante ? considerar tamb?m que diversos fatores n?o patol?gicos podem interferir em resultados dos exames hematol?gicos e bioqu?micos das aves, como a idade, sexo, per?odo reprodutivo, entre outros. O estudo teve como finalidade reconhecer e determinar padr?es hematol?gicos e bioqu?mico-s?ricos para a esp?cie em quest?o, elucidando poss?veis varia??es relacionadas a sexo, manejo e m?todo de conten??o. No experimento foram utilizados 68 exemplares, sendo 33 provenientes de um criat?rio comercial e 35 oriundos do acervo vivo da Funda??o RIOZOO. As amostras sangu?neas foram obtidas na veia jugular, transferidas, para tubos contendo EDTA e para outros sem anticoagulante. Somente as aves do RIOZOO foram tamb?m submetidas a uma nova coleta, que ocorreu 10 minutos ap?s anestesia com isoflurano, formando assim um grupo de 33 animais anestesiados e 35 n?o anestesiados. De cada animal foram aferidos trombocitometria, eritrocitometria, leucometrias global e espec?fica, determina??o de prote?nas plasm?ticas totais, ur?ia, creatinina, fosfatase alcalina, ?cido ?rico, colesterol, trigicer?deos, aspartato-aminotransferase, amilase, l?pase, creatinakinase, prote?nas totais s?ricas, albumina e globulinas. Os resultados foram analisados pelo teste ANOVA e teste t pareado. As f?meas apresentaram maior valor de trombocitometria. Os animais n?o anestesiados demonstraram um maior valor de leucometria global. N?o houve nenhuma diferen?a significatica entre machos e f?meas anestesiados. As aves oriundas do RIOZOO tiveram maior VG, hematimetria, VGM, contagem de eosin?filos e bas?filos, ur?ia, AST e CK, por?m, menores concentra??es de albumina e creatinina. O estudo revelou discrep?ncias a n?vel significativo entre os diferentes grupos estudados, permitindo um padr?o hematol?gico mais adequado com as caracter?sticas individuais de cada paciente.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2018-01-26T12:38:56Z No. of bitstreams: 1 2010 - Juberlan Silva Garcia.pdf: 6866273 bytes, checksum: 77b115c7b87deb998b7440f5ecb87940 (MD5)<br>Made available in DSpace on 2018-01-26T12:38:56Z (GMT). No. of bitstreams: 1 2010 - Juberlan Silva Garcia.pdf: 6866273 bytes, checksum: 77b115c7b87deb998b7440f5ecb87940 (MD5) Previous issue date: 2010-12-03<br>The trematode Echinostoma paraensei parasitizes the small intestine of rodents, being its natural definitive host Nectomys squamipes. In spite of the great importance of echinostomatides, there are few studies on morphology, biology and physiology of E. paraensei and its interaction with their intermediate and definitive hosts. In the present study 50 Rattus norvegicus (Wistar),adult females were used, 35 were individually infected with 150 E. paraensei metacercariae (Sumidouro strain) and 15 were maintained uninfected, as control group. Weekly, seven infected and three uninfected rodents were euthanized using CO2.The blood was collected to hematological analysis, and serum obtained by centrifugation and used to aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALKP), gamma-glutamyl transferase (GGT), bilirubin, glucose, total proteins and fractions determinations. Fragments of liver were collected to glycogen determination and histophatological analysis. Adult worms were loaded until third week of infection, being the higher number of worms loaded at first week of inection. The hepatocytes had rounded edges, with signs of impaired nuclear, mild infiltration of polymorphonuclear and sinusoids slightly expanded. The hepatocytes were vacuolated with discrete perivascular and periportal lymphocytic infiltration. The serum biochemical parameters analyzed were increased at the first week of infection, varying onward. Hematological analysis revealed the development of normocytic and normocromic anaemia with anisocytic alteration. In the white blood cells, only a leukopenia was observed at the third week of infection. The present results are discussed.<br>O tremat?deo Echinostoma paraensei ? um parasito de intestino delgado de roedores, sendo seu hospedeiro definitivo natural Nectomys squamipes. No presente estudo, foram utilizadas 50 f?meas adultas de Rattus norvegicus (Wistar), 35 animais infectados individualmente com 150 metacerc?rias de E. paraensei (linhagem Sumidouro) e 15 animais n?o infectados (controle). Semanalmente, sete animais infectados e tr?s animais controle foram submetidos ? eutan?sia com CO2. O sangue foi coletado para an?lise hematol?gica, e o soro obtido por centrifuga??o, para as determina??es de aspartato aminotransferase (AST), alanina aminotransferase (ALT), gamaglutamiltransferase (GGT), fosfatase alcalina (ALKP), bilirrubina, glicose, prote?nas totais e fra??es. Fragmentos de f?gado foram coletados para a determina??o de glicog?nio e para a an?lise histopatol?gica. Vermes adultos foram recuperados at? a terceira semana de infec??o, sendo o maior n?mro de vermes recuperados na primeira semana. Os hepat?citos apresentavam-se com bordos arredondados e alguns sinais de comprometimento nuclear, infiltra??o por polimorfonucleares nos sinus?ides, vacuoliza??o, discreta infiltra??o linfoplasmocit?ria perivascular e periportal com poucas c?lulas mononucleares presentes na luz do ducto biliar, alguns hepat?citos estavam hipotrofiados. As an?lises hematol?gicas revelaram o desenvolvimento de uma anemia normoc?tica normocr?mica, com anisocitose. N?o foram observadas altera??es significativas na s?rie branca, havendo apenas uma leucopenia na terceira semana de infec??o. Os resultados obtidos s?o discutidos.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-02T21:37:15Z No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5)<br>Made available in DSpace on 2017-05-02T21:37:15Z (GMT). No. of bitstreams: 1 2016 - Tatiane Cristina dos Santos Bonfim.pdf: 2465996 bytes, checksum: 94f74695ec8fd235bfbcd7f4a82aec87 (MD5) Previous issue date: 2016-02-25<br>CNPq<br>The interaction between intermediate hosts and helminths can cause metabolic and reproductive changes in the host snail, they start using their reserves to maintain its vital functions, immune system activation, repair of tissue damage and also to supply the necessary energy for the parasites development. Investigations of co-infection by different helminths can provide essential information about the biology of their co-existence. Our aims are investigate the reproductive changes, evaluate the activity of lactate dehydrogenase and the concentrations of glucose in the hemolymph and glycogen in the digestive gland complex and cephalopedal mass, and also verify histological and histochemical changes of Biomphalaria glabrata experimentally co-infected with Echinostoma paraensei and Angiostrongylus cantonensis. Five groups of snails i.e. uninfected, with single and double infections, either E. paraensei first (E+A) or A. cantonensis first (A+E) were followed; three times a week during four weeks the numbers of egg masses, eggs and hatched snails were counted. Histological and histochemical samples of the ovotestis, albumen gland, digestive gland complex and cephalopedal mass were collected after four weeks as well samples for the biochemical analysis. The number of egg masses/snail, eggs/snail and hatched snails showed significant differences comparing the control group to all infected groups, especially in group E + A, with the majority of values of parameters analyzed lower than 50% of those observed for control snails. A significant decrease in glucose levels and glycogen content, in contrast an increase of lactate dehydrogenase activity was observed in the infected snails. These results indicate intense energy demand that takes the snail increasing the anaerobic degradation of carbohydrates in the infected snails to obtain energy in an attempt to maintain homeostasis. The histological analysis showed that presence of both parasites in the all organs analyzed and the parasites were associated with cell disorganization.<br>A intera??o entre hospedeiros intermedi?rios e helmintos pode causar altera??es metab?licas e reprodutivas no molusco hospedeiro, que come?am a usar suas reservas para manter as fun??es vitais, na elabora??o de resposta imunol?gica e repara??o de danos teciduais e tamb?m para compensar a drenagem de nutrientes/energia pelos parasitos em desenvolvimento. Investiga??es sobre as co-infec??es por helmintos de esp?cies diferentes podem fornecer informa??es essenciais sobre a biologia da sua co-exist?ncia, refletindo aspectos mais pr?ximos ?queles que encontramos em condi??es naturais. Nossos objetivos foram investigar as altera??es reprodutivas, avaliar a atividade da lactato desidrogenase e as concentra??es de glicose na hemolinfa e glicog?nio no complexo gl?ndula digestiva-g?nada e massa cefalopediosa, e tamb?m verificar altera??es histol?gicas e histoqu?micas de Biomphalaria glabrata experimentalmente co-infectados com Echinostoma paraensei e Angiostrongylus cantonensis. Cinco grupos de moluscos (n?o infectados, com infec??es simples (Echinostoma paraensei ou Angiostrongylus cantonensis) e dupla, seja E. paraensei primeiro (E + A) ou A. cantonensis primeiro (A + E)) foram acompanhados; tr?s vezes por semana durante quatro semanas o n?mero de massas de ovos, n?mero de ovos e n?mero de moluscos eclodidos foram contados. Amostras para a an?lise histol?gica e histoqu?mica do ovoteste, gl?ndula do alb?men, complexo gl?ndula digestiva e massa cefalopediosa foram coletadas ap?s quatro semanas, assim como amostras para a an?lise bioqu?mica. O n?mero de massas de ovos/molusco, ovos/molusco e moluscos eclodidos dos moluscos infectados apresentaram diferen?as significativas quando comparados com o grupo controle, especialmente no grupo E + A, cuja a maioria dos valores dos par?metros analisados foi inferior a 50% dos valores observados para os moluscos controle. Um decr?scimo significativo nos n?veis de glicose e de glicog?nio, em contraste com um aumento de atividade da lactato desidrogenase foi observado nos exemplares infectados. Estes resultados indicam que a demanda de energia intensa leva o molusco infectado ao aumento da degrada??o anaer?bia de carboidratos para obten??o de energia buscando atender ao aumento da demanda energ?tica, numa tentativa de manter a homeostase glic?mica, por?m redu??es significativas s?o observadas. As an?lises histol?gicas e histoqu?micas mostraram a presen?a de ambos os parasitos nos ?rg?os analisados associados com intensa desorganiza??o celular.

Submitted by SBI Biblioteca Digital (sbi.bibliotecadigital@puc-campinas.edu.br) on 2018-02-15T12:50:05Z No. of bitstreams: 1 VANDERLEI JOSE JUNIOR.pdf: 1351825 bytes, checksum: 599d050ec951a6a8e7497a0cea5d2e17 (MD5)<br>Made available in DSpace on 2018-02-15T12:50:05Z (GMT). No. of bitstreams: 1 VANDERLEI JOSE JUNIOR.pdf: 1351825 bytes, checksum: 599d050ec951a6a8e7497a0cea5d2e17 (MD5) Previous issue date: 2017-12-15<br>Introduction: Prostate cancer is one of the most prevalent diseases in the male population, occupying the second position among malignant neoplasms. There are several therapeutic options for the treatment of localized prostate cancer, ranging from conservative behaviors to interventional treatments such as radical prostatectomy or external radiotherapy, associated or not with brachytherapy. Objective: To identify the factors that can predict biochemical recurrence and to evaluate treatment toxicity. METHOD: This is a retrospective and longitudinal study of 162 patients diagnosed with prostate cancer treated with conformational external radiotherapy associated with high dose rate brachytherapy (BTATD) between 2005 and 2014. The database was used of the Radium ? Campinas Oncology Institute, collected prospectively. Results: The mean follow-up time was 57 months. No grade 3 late toxicity was observed in the gastrointestinal tract, with only 1 patient (0.6%) genitourinary tract. The only categorical variable that presented statistical significance for biochemical relapse was the Nadir PSA <1 ng / ml (p = 0.018). The biochemical recurrence rate found was 96.3%, based on the Phoenix criteria (PSA nadir + 2 ng / ml). Conclusions: This study demonstrated that in the treatment of localized prostate cancer, the association of external radiotherapy with BATD is a safe therapeutic option, with a low degree 3 late toxicity and a biochemical recurrence of only 3.7% (with HF = 95 %).<br>Introdu??o: O c?ncer de pr?stata ? uma das doen?as mais prevalentes na popula??o masculina, ocupando a segunda posi??o entre as neoplasias malignas. H? v?rias op??es terap?uticas para o tratamento do c?ncer de pr?stata localizado, podendo variar de condutas conservadoras ? tratamentos intervencionistas como a prostatectomia radical ou a radioterapia externa, associada ou n?o ? braquiterapia. Objetivo: Identificar os fatores que possam predizer recidiva bioqu?mica e avaliar a toxicidade do tratamento. M?todo: Tratase de um estudo retrospectivo e longitudinal, com 162 pacientes diagnosticados com c?ncer de pr?stata, tratados com radioterapia externa conformacional associada ? braquiterapia de alta taxa de dose (BATD), entre 2005 e 2014. Utilizou-se o banco de dados do Radium - Instituto de Oncologia de Campinas, coletados prospectivamente. Resultados: O tempo m?dio de seguimento foi de 57 meses. N?o foi observada toxicidade tardia grau 3 no trato gastrointestinal, sendo apenas 1 paciente (0,6%) trato genitourin?rio. A ?nica vari?vel categ?rica que apresentou signific?ncia estat?stica para recidiva bioqu?mica foi o PSA Nadir <1 ng/ ml (p = 0,018). A taxa de recidiva bioqu?mica encontrada foi de 96,3%, baseando-se nos crit?rios de Phoenix (PSA nadir + 2 ng/ml). Conclus?es: Esse estudo demonstrou que, no tratamento de c?ncer de pr?stata localizado, a associa??o de radioterapia externa com BATD ? uma op??o terap?utica segura, com baixa taxa de toxicidade tardia grau 3 e recidiva bioqu?mica de apenas 3,7% (com I.C = 95%).

碩士<br>國立體育大學<br>運動與健康科學學院<br>105<br>Electronic cigarettes (EC) are becoming a preferred replace for nicotine delivery among many smokers in recent year. In some research support that EC could be useful for smoking cessation. However, EC effects of on human health are inconclusive due to lack of empirical research investigating the presence of EC-induced health hazards or benefits. Thus, we examined the effect of vapor produced by EC on exercise performance and health-related profile in mice model. Female ICR mice were divided into 5 groups (n = 6 per group) for 14 days with no intervention, exposure in air, 0, 0.6mg/mL and 6mg/mL nicotine designated control, Air, ELN-0X, ELN‐1X and ELN‐10X groups, respectively. The results indicated that the EC decreased the grip strength (p = 0.0024) with ELN-10X. EC treatments also produced dose-dependent decreases in liver and muscle glycogen storage (p = 0.0009; p = 0.0003) with ELN-10X. In addition, EC treatment had no negative effect on levels of biochemical indices. We also found the major organ had no adverse effect on morphology. Therefore, we suggest that after use EC could reduce explosive force and glycogen storage capacity. In addition, we could not found any negative effect on biochemical profile and organ morphology.

碩士<br>國立體育大學<br>運動與健康科學學院<br>102<br>Abstract Purpose: To investigate the effects of whole-body vibration training (WBV) on physiological and biochemical characteristics in a mouse model. Methods: Twenty-four male C57BL/6 mice age 4 wk were divided randomly into three groups (n=8 per group): (i) sedentary control (SC), (ii) WBV at relative low-intensity (5.6 Hz, 0.13 g) (VL), or (iii) relative high-intensity (13 Hz, 0.68 g) WBV (VH). The exercise performance and exercise-induced fatigue test were evaluated using forelimb grip strength, endurance swimming time, and levels of serum lactate, ammonia, glucose, and creatine kinase (CK) after a 15-min swimming exercise. We also measured the body composition and biochemical parameters of test animals at the end of the experiment. Results: A trend analysis revealed that WBV interventions increased the grip strength and aerobic endurance. WBV produced dose-dependent decreases in serum lactate, ammonia and glucose levels after the 15-min swimming test. WBV also produced decreases in relative heart and liver weights in a dose-dependent manner, as well as fasting serum levels of aminotransferases, CK and blood urea nitrogen. Conclusion: These findings suggest that WBV can improve exercise performance, anti-fatigue, and and has no adverse side effects on clinical biochemical assessments in health mice. Therefore, we suggest that WBV may be an effective intervention for the improvement of health promotion and physical performance in general adult population.

Dissertação de Mestrado em Biologia Celular e Molecular apresentada à Faculdade de Ciências e Tecnologia<br>A plasticidade sináptica que ocorre ao nível das sinapses excitatórias é um dos mecanismos cerebrais mais importantes para um normal desenvolvimento da nossa capacidade intelectual, sendo essencial para todas as funções cognitivas. O glutamato é o neurotransmissor mais abundante no sistema nervoso central e o principal mediador da neurotransmissão excitatória. A ação deste neurotransmissor é dependente da sua ligação a recetores na membrana pós-sináptica, podendo atuar em dois tipos de recetores: inotrópicos, que quando ativos permitem a passagem de iões como Na+, K+ e Ca2+; e metabotrópicos, cuja função é ativar cascatas de proteínas intracelulares. A ação conjunta destes recetores modula o funcionamento de grande parte dos circuitos neuronais essenciais para as funções cognitivas. Assim, todos os constituintes do complexo sistema que forma as sinapses gluatamatérgicas desempenham um papel essencial nas regiões cerebrais que maioritariamente regulam a capacidade de aprendizagem e formação de memórias, tais como o hipocampo, o córtex pré-frontal e o cerebelo.Os recetores ionotrópicos de glutamato do tipo AMPA são os principais mediadores da neurotransmissão excitatória rápida. A eficiência da transmissão glutamatérgica é dependente do padrão de atividade neuronal, podendo ser reforçada ou enfraquecida dependendo da quantidade de recetores na membrana celular sináptica, fenómeno designado de plasticidade sináptica. Os recetores AMPA interagem com várias proteínas que influenciam a sua funcionalidade. Entre estas destaca-se a família de proteínas chamadas TARP (proteínas transmembranares associadas aos recetores AMPA). Estas proteínas regulam a atividade dos recetores a vários níveis, tais como modulando o seu tráfego sináptico e as propriedades biofísicas do canal iónico dos recetores. Dentro desta família de proteínas, a stargazina é a mais conhecida e uma das mais abundantes. Esta proteína tem como principal função estabilizar os recetores na membrana sináptica, de forma dependente da sua interação com a PSD95. Tendo em conta o seu importante papel na regulação da atividade dos recetores AMPA, a stargazina é essencial para o correto funcionamento dos processos de plasticidade sináptica. Esta proteína é codificada pelo gene CACNG2, e murganhos que possuem uma forma corrompida deste gene (murganhos stargazer) têm uma redução dramática na atividade neuronal dependente dos recetores AMPA nas regiões cerebrais onde a proteína é mais abundante.Vários estudos anteriores têm sugerido que a disfunção das sinapses glutamatérgicas está implicada em vários distúrbios psiquiátricos e do desenvolvimento, tais como o défice cognitivo, distúrbios do espectro autista e esquizofrenia. A anormal funcionalidade destas sinapses é frequentemente associada a anormalidades nas proteínas sinápticas como as TARPs. Este projeto teve como objetivo estudar uma mutação (p. Val143Leu) no gene CACNG2 que foi identificada numa criança com défice cognitivo. Um estudo anterior revelou que esta mutação diminui na capacidade da stargazina para interagir com os recetores AMPA, levando a uma diminuição dos níveis dos recetores na sinapse em neurónios transfetados para expressarem esta forma mutada. Foi criado pelo nosso laboratório um modelo animal de murganho geneticamente modificado que expressa a stargazina mutada (stargazina V143L). O estudo aqui apresentado descreve a caracterização deste animal modelo, ao nível da composição bioquímica das sinapses, morfologia dos neurónios do hipocampo e comportamento social, realizada com o intuito de tentar perceber de que forma a mutação na stargazina altera a capacidade funcional da proteína e como pode levar a um fenótipo característico de distúrbios como o défice cognitivo. Descobrimos que os neurónios da região CA1 do hipocampo destes animais têm uma arborização dendrítica de complexidade reduzida, bem como uma redução no tamanho das suas dendrites. Para além disso observámos que a forma mutada da stargazina se encontra menos expressa na sinapse. Finalmente, verificamos que estes animais exibem um comportamento social anormal e alterações no comportamento perseverante de ocultação de objetos, típico dos murganhos. Concluindo, este estudo demonstra uma relação de causalidade entre a mutação V143L na stargazina, detetada num doente com défice intelectual, e alterações na morfologia neuronal no hipocampo e no comportamento social de murganhos. Contudo, é necessária uma análise mais aprofundada de modo a perceber em que medida é que esta mutação afeta a função da stargazina, levando consequentemente a alterações nos circuitos neuronais que regulam os comportamentos afetados nos animais. Com os resultados provenientes destas análises esperamos perceber melhor como alterações genéticas desta ordem levam a este tipo distúrbios cognitivos. A plasticidade sináptica que ocorre ao nível das sinapses excitatórias é um dos mecanismos cerebrais mais importantes para um normal desenvolvimento da nossa capaci<br>The plasticity of excitatory synapses is an essential brain process involved in cognitive functions and necessary for the proper development of intellectual capacities. Glutamate, the major excitatory neurotransmitter in the central nervous system, acts on ionotropic and metabotropic receptors. Together and distinctly, these receptors modulate neuronal circuits that underlie aspects of cognitive function. Therefore, glutamatergic synaptic transmission is a key player in learning and memory formation processes in brain areas such as the hippocampus, the prefrontal cortex and the cerebellum.Within glutamatergic synapses AMPA receptors (AMPARs) mediate most of the rapid excitatory neurotransmission and undergo activity-dependent changes in their trafficking and surface expression, which have been proven to be fundamental mechanisms for synaptic plasticity processes such as long-term potentiation (LTP). An increase in the post-synaptic response to a stimulus is achieved either through elevating the number of AMPARs at the post-synaptic surface or by increasing the single channel conductance of these receptors. The ability to control the surface expression and activity of AMPARs is also essential for homeostatic plasticity, a set of mechanisms that act in order to stabilize neuronal and circuit activity by counterbalancing some of the plastic challenges faced by neurons. AMPARs directly interact with transmembrane AMPA receptor regulator proteins (TARPs), which act as their auxiliary subunits. TARPs influence AMPAR synaptic targeting, synapse expression and function by different mechanisms. Among the TARPs, stargazin (γ-2) is one of the most important and abundant members of this family. This protein regulates AMPA receptor function and stabilizes AMPARs in the synaptic membrane due to a strong interaction with PSD-95; consequently, it plays a crucial role in synaptic plasticity. Stargazin is encoded by the human CACNG2 gene, and mice in which the homologous gene is disrupted (stargazer mice) show a dramatic loss of AMPA receptor activity in brain regions where stargazin is highly expressed.A significant amount of previous studies suggest that the dysfunction of glutamatergic synapses is strongly implicated in several neurodevelopmental disorders, such as Intellectual disability (ID), autism spectrum disorders and schizophrenia. The disruption of glutamatergic synapses is often related with the dysfunction of synaptic scaffold proteins or TARPs. In this study, we looked into a de novo mutation (p. Val143Leu) in the CACNG2 gene that was identified in a male with moderate ID. A previous study showed that this mutation significantly decreases stargazin's ability to bind to AMPARs and reduces cell surface expression of the GluA1 AMPAR subunit in transfected hippocampal neurons and HEK293 cells. In order to elucidate how this mutation affects protein function and contributes to the development of disease-associated phenotypes our laboratory generated a knock-in mouse harboring the human mutation in the stargazin-encoding gene. In this project, we characterized the synaptic biochemical composition, hippocampal neuronal morphology and social behavior featured by these mice, to address the role of stargazin in normal neuronal development and to determine causality between a disease-associated mutation in the CACNG2 gene and the generation of ID-like behavior in mice. Here, we found that mutant stargazin levels are decreased at whole-brain derived postsynaptic densities, but found no evidence for alterations in AMPARs subunits synaptic expression. Remarkably, neuronal morphology analyses revealed that stargazin V143L+/+ knock-in mice CA1 pyramidal neurons exhibit decreased dendritic arborization complexity and a decrease in the total length of dendrites. Finally, stargazin V143L+/+ knock-in mice displayed abnormal social behavior in the three chamber test and impairment in the perseverative species-typical burying behavior.In conclusion, this study revealed that the altered form of stargazin, resultant from the V143L mutation in the CACNG2 gene, is indeed causative of morphological and behavioral abnormalities in mice. Further analyses are needed to complete the characterization of this mouse model and to better understand to what extent the ID-associated mutation affects stargazin function, and leads to neuronal circuits alteration that underlie the behavior impairments reported in this study. Ultimately, we hope that the knowledge resulting from the study of these mice gives us valuable insights to understand the big picture of the stargazin-associated disorders and other similar conditions.The plasticity of excitatory synapses is an essential brain process involved in cognitive functions and necessary for the proper development of intellectual capacities. Glutamate, the major excitatory neurotransmitter in the central nervous system, acts on ionotropic and metabotropic receptors. Together and distinctly, these r

碩士<br>國立中興大學<br>生命科學院碩士在職專班<br>105<br>Psoriasis is an immune-mediated chronic skin disorder. It affects about 125 million people worldwide. The prevalence of psoriasis in the adult population is estimated at 0.91% in the United States to 8.5% in Norway. The characteristics of psoriasis include abnormal hyperplasia in epidermis, production of plaques and inflamed dander, and immune cell infiltration symptoms. Although psoriasis is not an infectious disease, social and quality of life are affected in patients with psoriasis. At present, the real etiology of psoriasis is still unclear, but may relate to a number of factors. Imiquimod (IMQ), a toll-like receptor 7 (TLR7) ligand, is a potent immune-modulatory agent. Studies have indicated that topical application of IMQ could induce psoriasis-like skin inflammation in mice. Recent studies have indicated some blood or biochemical profile, such as mean platelet volume (MPV)、platelet distribution width (PDW) and C-reactive protein (CRP) are significant changes in psoriasis patients. In this study, the blood and biochemical profile of mice with IMQ-induced psoriasis-like skin inflammation were examined. We observed that MPV, PDW, mean corpuscular volume (MCV), red cell distribution width standard (RDW-SD), red cell distribution width coefficient of variation (RDW-CV) and CRP are increase. In contrast, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), biochemical projects cholesterol (Cho), low-density lipoprotein (LDL), High-density lipoprotein (HDL) and Albumin (Alb) are significantly decrease. These changes are similar to blood and biochemistry profiles of human psoriasis. Thus, IMQ not only induces mice with psoriasis-like skin inflammation which similar human psoriasis, the blood and biochemical changes are also similar to human psoriasis.

碩士<br>國立中正大學<br>分子生物研究所<br>96<br>Many gene products that are essential to pancreas development, such as Pdx-1、Ptf1a、Ngn3 and Nkx family, generally show strong expression specifically in developing pancreas. Previously, a protein with RNA binding motif, named RBMS3, was found to be expressed strongly in early embryonic pancreas. Although RBMS3 was first identified and cloned by screening of a fibroblast expression library with the labeled DNA fragment derived from the promoter of the mouse collagen ?(I) gene, its specific presence in developing pancreas has been confirmed by in situ hybridization and immunofluorescence staining. In order to study the function of RBMS3, a yeast two-hybrid system had been applied to identify RBMS3-interacting proteins, and several genes including SPIN3 (Spindlin 3), DUSP14 (dual specificity phosphatase 14) were identified from the screening. I have confirmed the interactions between SPIN3 and DUSP14 with RBMS3 by pull down assay and co-immunoprecipitation. In addition, I have generated a transgenic construct that contains the RBMS3 cDNA under the control of Pdx-1 promoter. This construct had been used to generate transgenic mice that overexpress RBMS3 in developing pancreas. I have measured blood sugar of fasted transgenic mice when they were 6 weeks old. However, there was no significant difference between wild and transgenic mice. I have also attempted to observe differences in cell-type composition in the embryonic, postnatal and adult pancreas. However, no apparent influence of the ectopic RBMS3 was observed on these transgenic mice. It is likely RBMS3 modulates pancreas development with a more subtle mechanism. The results described above may provide some information of RNA-binding protein’s function and regulation in pancreas development. This is a beginning for researchers to note the role of RNA-binding proteins in organogenesis.

碩士<br>國立臺灣大學<br>生物化學研究所<br>77<br>Aldrich 公司生產的粗製腐植酸（crude humic acid）經Sephadex G-25 膠體管柱分 離得到純化的G25-A 腐植酸。把純化的G25-A 腐植酸與陽離子群製作成「複合物」。 每天以0.125 毫克/0.1毫升的複合物劑量，腹腔注射雄性小白鼠。當注射到第４０天 時，發現小白鼠肝、腎細胞的肉鹼－棕櫚醯轉移（carnitine palmitoyltransfer- ase） 及肉鹼－乙醯轉移（carnitine acetyltransferase） 活性增加，若是只有 單獨的G25-A 腐植酸或是單獨的陽離子群則不能影響這兩種酵素活性；複合物也會降 低肝細胞內過氧化氫（Catalase）的活性。同樣的，單獨的G25-A 腐植酸或是單獨 的陽離子群也不會影響肝細胞內過氧化氫的活性。至於腎細胞內過氧化氫的活性 ，則不受複合物之影響。 由電泳分析（PAGE）顯示：複合物會使肝細胞漿液中分子量為60kDa 的多胜（pol- ypeptide）消失，而腎細胞內的60kDa 多胜仍然存在。 從組織病理切片可以看見細胞受複合物的影響後發生中度脂肪性改變（mild fatty c hanges），細胞內有退化核，甚至細胞體積增大（hypertrophy），細胞變性（degen eration）等現象。

The ability to extrapolate nutritional intervention data from experimental rodent models to humans requires standardization of dietary design. The inability to translate the level of nutrients from animal models to humans has contributed to contradictory findings between species. It is hypothesized that dietary linoleic acid (LA) promotes chronic and acute diseases by enriching tissues with arachidonic acid (AA), its downstream metabolite. However, levels of LA in rodent diets are notoriously erratic making interspecies comparisons unreliable. Therefore, the ability to extrapolate the biological effects of dietary LA from experimental rodents to humans necessitates an allometric scaling model that is rooted within a human equivalent context. To determine the physiological effect of dietary LA on tissue AA, a mathematical model for extrapolating nutrients based on energy was designed to mimic human equivalent doses. C57BL/6J mice were divided into 9 groups fed a background diet equivalent to that of the US diet (including LA, ALA, AA, EPA, DHA) with supplemental doses of LA (up to 2.3x) or AA (up to 5x). Changes in the phospholipid fatty acid compositions were monitored in plasma and erythrocytes and compared to data from humans supplemented with equivalent doses of LA or AA. Increasing dietary LA had little effect on tissue AA, while supplementing diets with AA significantly increased tissue AA levels, recapitulating results from human trials. Thus, interspecies comparisons for dietary LA between rodents and humans can be achieved when rodents are provided human equivalent doses based on differences in metabolic activity as defined by energy consumption.

博士<br>靜宜大學<br>食品營養學系<br>99<br>This dissertation aimed to investigate the biochemical properties of β-N-acetylhexosaminidase (β-NAHA) from fig latex and chitosanase from bamboo shoots, as well as the safety evaluation and effect of low molecular weight chitosan (LMWC), prepared by bamboo chitosanase, on mice with renal lesions. Four studies were conducted to reach the goals of researches. First, a major β-NAHA was purified from fig latex. Biochemical properties, including molecular mass, isoelectric point, thermal stability, effectors, and substrate specificity, of the purified β-NAHA and kinetics of competition for the enzyme with mixed substrates were studied. The results indicated that the activity of the fig β-NAHA was specific for the β-glycoside linkage, and the enzyme had only one active site for binding both substrates p-nitrophenyl-N-acetyl-β-D-glucosaminide and p-nitrophenyl-N-acetyl-β-D-galactosaminide. In the second part of study, a predominant chitosanase induced by chitosan was isolated and purified from edible bamboo shoots. The biochemical properties of the purified chitosanase were studied as well. The results indicated that the molecular mass of the purified chitosanase was 12.7 kDa, and this enzyme was capable of hydrolyzing chitosan polymers with various degree of deacetylation (10.9-94%) and the chitosan with ~30% deacetyled was the most susceptible. In the third part of study, a LMWC (~29 kDa) was prepared by the depolymerzation of chitosan using partially purified bamboo chitosanase. Animal experiments showed that the LMWC did not cause damage of nucleus in mice and had no toxicity in rats, and the no-observed-adverse-effect level (NOAEL) was greater than 1 g/kgBW in rats. In the fourth part of study, renal lesions were induced by aristolochic acid (AA) in mice and then the repairing effect of LMWC on renal lesions was studied. The results of biochemical analysis and histophathological examination indicated that oral administration of LMWC at a middle dose of 500 mg/kg BW/day for 14 days effectively ameliorated AA-induced renal lesions in mice.

Surface water pollution is prevalent in numerous areas of central Roodepoort mainly due to gold mining activities. The surface water quality for the Bosmontspruit, Russell’s Stream and the New Canada Dam was assessed from October 2010 to March 2011. Physical, chemical and biological characteristics of the water were determined for 8 monitoring points and the results obtained were compared with the In-stream water quality guidelines for the Klip River catchment and the South African Water Quality Guidelines. A trend noticed throughout the sampling period was the non-compliance in the levels of total dissolved solids (TDS) and dissolved oxygen. The results indicated that concentrations of iron, aluminium, nickel, manganese and potassium were above the limit across the Bosmontspruit and Russell’s stream. There was also significant evidence of excessive faecal coliform and ammonium pollution in the Bosmontspruit. During the monitoring period it was noted that water from these streams were utilised for crop irrigation, bathing, livestock and human consumption and may pose a health hazard due to poor water quality.<br>Environmental Sciences<br>M.Sc. (Environmental Science)