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Characterization of the human ceruloplasmin cDNA and geneKoschinsky, Marlys Laverne January 1988 (has links)
A cDNA for human ceruloplasmin was identified in a human liver cDNA library by screening with mixtures of synthetic oligonucleotides complementary to two regions of the ceruloplasmin mRNA. The resulting clone (phCP-1) contained DNA coding for amino acid residues 202 - 1046 of the protein, followed by a 3' untranslated region of 123 bp and a poly(A) tail. To isolate additional clones extending in a 5' direction, two randomly-primed human liver cDNA libraries were constructed in the bacteriophage vectors λgt10 and λgtll. From the former library, a clone was isolated (λhCP-1) that contained DNA coding for a putative signal peptide consisting of 19 amino acid residues, followed by DNA encoding residues 1 - 380 of plasma ceruloplasmin. From the λgtll library, six ceruloplasmin cDNA clones were purified, two of which were shown to contain 10 and 38 bp of non-coding sequence extending 5' to λhCP-1. Blot hybridization analysis using cDNA probes showed that ceruloplasmin mRNA from the human hepatoma cell line HepG2 is 3700 nucleotides in size, while human liver RNA contained an additional hybridizing species of 4500 nucleotides in size.
Ceruloplasmin genomic DNA clones (spanning a region of approximately 45 Kbp) were obtained by the screening of several human genomic phage libraries using cDNA probes. These clones were initially characterized by restriction endonuclease mapping. Using DNA sequence analysis, the positions of intron/exon boundaries were determined. To date, 14 exons (average size of 183 bp) have been identified in the ceruloplasmin gene, corresponding to nucleotide residues 1 - 2565 of the coding sequence. The majority of the 14 introns localized within this region were located in analogous positions in the factor VIII gene, thereby suggesting that these two proteins have evolved from a common ancestral gene.
At least 4 exons have been localized within the 5' untranslated region of the human ceruloplasmin gene, although typical eukaryotic promoter elements have not yet been identified. The significance of this novel organization remains unclear at present.
In addition to the wild-type gene, a processed pseudogene for human ceruloplasmin was identified and contained DNA corresponding to the functional gene sequence encoding the carboxy-terminal 563 amino acid residues and the 3' untranslated region. The pseudogene appears to have arisen from a processed RNA species, since intervening sequences coincident with those of the functional gene have been removed with the exception of a short segment of intronic sequence which denotes the 5' boundary of the pseudogene. Based on genomic Southern blot anlysis performed under high stringency conditions, the pseudogene seems to comprise the only sequence in the human genome that is closely related to the wild-type gene. Using somatic cell hybridization, the pseudogene was localized to human chromosome 8; this differs from the location of the wild-type ceruloplasmin gene, which has been mapped to chromosome 3. / Medicine, Faculty of / Biochemistry and Molecular Biology, Department of / Graduate
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Genetic and environmental influences on the serum protein ceruloplasmin.Cox, Elizabeth Diane Wilson. January 1968 (has links)
No description available.
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Physical-chemical and immunochemical properties of human ceruloplasmin and some of its derivativesKasper, Charles Boyer, January 1962 (has links)
Thesis (Ph. D.)--University of Wisconsin--Madison, 1962. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 240-248).
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Studies of immunoglobulin impurities and of the auticomplementary activity of crystalline ceruloplasminFisher, Gwendolyn Barbara, January 1969 (has links)
Thesis (M.S.)--University of Wisconsin--Madison, 1969. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.
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Genetic and environmental influences on the serum protein ceruloplasmin.Cox, Elizabeth Diane Wilson. January 1968 (has links)
No description available.
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Influence of ascorbic acid supplementation on copper status in young adult menFinley, Elizabeth Bidwell 14 August 1981 (has links)
Thirteen healthy adult males, ages 20-40, consuming self selected
diets, were given instructions to take one 500 mg tablet of ascorbic
acid three times a day with their meals for a period of ten weeks. The
effect of this daily supplementation on copper status was investigated.
An estimation made from a three day diet record kept by each subject
indicated their dietary copper intake to be 1.92 mg per day. Determination
of serum ceruloplasmin and serum copper done on the first day
of the ascorbic acid supplementation period showed that the subjects
fell within accepted ranges of normal. All further determinations of
these parameters during the experimental period were compared to initial
values so that each subject served as his own control.
At week seven the high ascorbic acid intake significantly decreased
ceruloplasmin by 26 percent. At the end of the ten week ascorbic acid
supplementation period, serum ceruloplasmin activity was significantly
lowered by 20 percent. The slight increase over week seven was attributed
to a drop in compliance to taking the ascorbic acid tablets.
Serum copper levels were not significantly affected during the 10 week
experimental period although a consistent decrease was observed. Two
weeks after acerbic acid was terminated serum ceruloplasmin activity
increased but was not significantly different from week ten values.
However, when compared to week seven values, a significant increase
of 14 percent was observed. Serum copper levels measured two weeks
after ascorbic acid supplementation was terminated significantly increased
14 percent over week ten values.
The results of this human volunteer study indicate that taking
a megadose of ascorbic acid for ten weeks will significantly decrease
serum ceruloplasmin activity much like that observed in laboratory
animal studies. Based on this finding, one may question the safety
of prolonged self-dosage of high amounts of ascorbic acid by adults
as encouraged by the popular press. / Graduation date: 1982
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Avaliação de fontes de cobre para ovinos com ensaio de biodisponibilidade / Influence of diferente levels and sources of copper supplementation with bioavailability studyYoshikawa, Carolina Yumi Cascão 21 March 2014 (has links)
O objetivo do estudo foi estimar a biodisponibilidade de duas fontes de cobre: orgânica (cobre metionina) e inorgânica (sulfato de cobre) na dieta de cordeiros. O experimento foi conduzindo na FZEA USP de Pirassununga utilizando 40 cordeiros da raça Merino X Texel, que foram distribuídos aleatoriamente em cinco grupos, e submetidos a cinco tratamentos, totalizando oito animais por tratamento: Tratamento 0: Dieta controle sem adição de Cu; Tratamento 1: 10 mg de Cu/Kg de MS na forma de sulfato de Cu; Tratamento 2: 30g de Cu/Kg de MS na forma de sulfato de Cu; Tratamento 3: 10 mg de cu/kg de MS na forma de cobre metionina; Tratamento 4: 30 mg de cu/kg de MS na forma de cobre metionina. Foram feitas biópsias do fígado dos animais no tempo zero para análise de cobre e colhidas amostras de sangue nos dias 0, 28, 56 e 84 dias para determinação de Cu sérico, atividade de ceruloplasmina e enzimas de função hepática. Ao final do experimento, os animais foram abatidos para colheita de amostras de fígado, músculo e rim, para determinação dos teores de Cu e da enzima superóxido dismutase (SOD). Nos últimos dez dias do experimento foi realizado um balanço metabólico de cobre. A biodisponibilidade foi calculada pela técnica \"slope ratio\", utilizando como parâmetros a concentração de cobre no fígado. Não houve diferença (P>0,05) no desempenho dos animais (peso vivo e ganho de peso) entre os tratamentos. A concentração sérica de AST e ALT permaneceu abaixo dos níveis de intoxicação em todos os tratamentos, durante todo o período. A atividade da ceruloplasmina não diferiu entre os tratamentos (P>0,05). O teor de cobre no soro, na biópsia do fígado e no músculo não foi diferente (P>0,05) entre os tratamentos. Entretanto, a concentração do mineral no fígado dos animais suplementados (284,28 mg/kg) foi maior (P<0,05), quando comparados ao grupo controle (168,01 mg/kg), assim como o Cu-met 30 mg/kg (341,29 mg/kg) foi superior (P<0,05) ao de 10mg/kg MS (263,02 mg/kg). A atividade da SOD nos animais suplementados (µmol/mg prot foi superior à do grupo controle. Nos rins o teor de cobre foi superior nos animais que receberam 30mg/kg de MS de Cu-met (6,65 mg/kg) em relação aos que receberam 10 mg/kg de MS da mesma fonte (3,86 mg/kg). A absorção e a retenção aparentes do cobre foram maiores para a fonte inorgânica, comparada com a orgânica. A biodisponibilidade do cobre determinada pela concentração de cobre no fígado, utilizando a técnica do \"slope ratio\", considerando o CuSO4 como padrão (100%), apresentou disponibilidade de 150,64% para o Cu-met. / The study was conducted to estimate the relative bioavailability of two sources of supplemental copper: organic (copper methionine) and inorganic (copper sulfate) in the diet of lambs, by analyzing the concentration of copper and enzymes in the liver and metabolic balance calculation, using 40 lambs breed Merino X Texel, which was fed three concentrations of copper (basal + two additions) in two sources, which were randomly allotted to five groups, and subjected to five treatments: treatment 0: control (diet without addition of Cu); treatment 1: (diet with 10 mg Cu/Kg DM of CuSO4); Treatment 2: (diet with 30 g Cu/Kg DM of CuSO4); Treatment 3: (diet with 10 mg Cu/kg DM of copper methionine; Treatment 4: (diet with 30 mg cu/kg DM of copper methionine). Liver biopsies were made on 0 d. Blood samples were taken via the jugular vein on 0, 28, 56 and 84 d to determine serum Cu and serum ceruloplasmin and liver transaminases (AST, ALT) concentrations. The animals were slaughtered and samples of liver, kidney and muscle were taken for the determination of the levels of Cu and superoxide dismutase activity. In the last ten days of the experiment a metabolic balance of copper was conducted. The bioavailability was calculated by the \"slope ratio\" technique, using the concentration of copper in the liver as parameter. There was no difference (P >0.05) on animal performance (live weight and weight gain) among treatments. The serum AST and ALT levels remained below poisoning in all treatments during the period. The ceruloplasmin activity did not differ between treatments (P>0.05). The copper content in biopsy, serum and muscle was not different (P>0.05) between treatments. However, the mineral concentration in the liver of animals fed (284.28 mg/kg) was higher (P <0.05) when compared to the control group (168.01 mg/kg ) and 30 Cu -met mg/kg (341.29 mg/kg) was higher (P <0.05) at 10mg/kg MS (263.02 mg/kg). The SOD activity in the supplemented animals (mmol/mg prot) was superior to the control group. Copper in Kidneys was higher in animals that received 30mg/kg MS meth-Cu (6.65mg/kg) compared those receiving 10 mg/kg DM from the same source (3.86 mg/kg). Apparent absorption and retention of copper were higher for inorganic source, compared with the organic. The bioavailability determined by the concentration of copper in the copper liver, using the technique of \"slope ratio\", considering CuSO4 as standard ( 100% ) presented availability of 150.64 % for Cu-met.
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Efeitos de fontes orgânicas de cobre e enxofre sobre a interação cobre, molibdênio e enxofre na alimentação de ovinos / Effects of organic sources of copper and sulfur on interaction copper, molybdenum and sulfur in sheep feedingConti, Renata Maria Consentino 17 December 2014 (has links)
O presente trabalho teve como objetivo estudar os efeitos que as fontes orgânicas e inorgânicas de cobre e enxofre possuem na interação cobre-enxofre-molibdênio, estimando a biodisponibilidade de duas fontes de cobre na dieta de ovinos. Para isso, foram utilizados 40 ovinos desmamados, com aproximadamente 3 meses e peso médio 20 kg, distribuídos em 10 tratamentos, sendo: 1) dieta basal; 2) dieta basal contendo 10 mg de molibdênio/kg de MS; 3) dieta basal + 10 mg cobre inorgânico/kg de MS + 0,2% enxofre inorgânico/kg de MS; 4) dieta basal + 10 mg cobre inorgânico/kg de MS + 0,2% enxofre orgânico/kg de MS; 5) dieta basal + 10 mg cobre orgânico/kg de MS + 0,2% enxofre inorgânico/kg de MS; 6) dieta basal + 10 mg cobre orgânico/kg de MS + 0,2% enxofre orgânico/kg de MS; 7) dieta com 10 mg molibdênio + 10 mg cobre inorgânico/kg de MS + 0,2% enxofre inorgânico/kg de MS; 8) dieta com 10 mg molibdênio + 10 mg cobre inorgânico/kg de MS + 0,2% enxofre orgânico/kg de MS; 9) dieta com 10 mg molibdênio + 10 mg cobre orgânico/kg de MS + 0,2% enxofre inorgânico/kg de MS; 10) dieta com 10 mg molibdênio + 10 mg cobre orgânico/kg de MS + 0,2% enxofre orgânico/kg de MS. O experimento teve duração total de 84 dias, sendo realizadas pesagens dos animais nos dias 1, 28, 56 e 84 dias para acompanhamento do desenvolvimento. Foram realizadas também coletas sanguíneas para estudo de teores de cobre, enxofre e molibdênio sanguíneos, teores de glicose, albumina, ureia, colesterol, triglicerídeos, hematócrito e ceruloplasmina. Foram realizadas biópsias do fígado no tempo zero para análises de cobre, enxofre e molibdênio. Ao término do período experimental os animais foram abatidos e colheu-se amostras de fígado e líquido biliar para determinação final dos teores de minerais, bem como acompanhamento do pH e peso das carcaças quente e fria. No terço final do período experimental foi realizado um balanço metabólico para cobre, enxofre e molibdênio, sendo a biodisponibilidade do cobre calculada pela técnica \"slope ratio\", utilizando-se os teores de cobre hepático. Os parâmetros foram analisados considerando-se a existência de uma estrutura de tratamento fatorial 2 x 2 x 2 (com e sem molibdênio, cobre orgânico e inorgânico e enxofre orgânico e inorgânico) e uma dieta basal e uma basal mais molibdênio, em delineamento inteiramente casualizado. As concentrações de glicose, albumina, ureia, colesterol e hematócrito sanguíneos não sofreram efeito (P>0,05) pela adição de molibdênio ou adição de cobre e enxofre, ou pela interação cobre-enxofre-molibdênio. Entretanto as concentrações de triglicerídeos sanguíneos foram alteradas (P<0,05) pela interação cobre-enxofre, mostrando redução na sua concentração quando da adição de cobre e enxofre nas fontes orgânicas. O peso vivo dos ovinos não foi influenciado (P>0,05) pelos tratamentos, porém o ganho de peso foi influenciado pela interação cobre-enxofre-molibdênio onde se observou redução no ganho de peso com a adição de molibdênio exclusivo e aumento no ganho de peso quando adicionado molibdênio com cobre e enxofre nas fontes orgânicas quando comparados com as fontes inorgânicas. A suplementação de fontes de cobre e enxofre e adição de molibdênio não influenciaram (P>0,05) os valores de pH e peso das carcaças. A presença de molibdênio na dieta diminuiu os teores de ceruloplasmina e a presença e molibdênio exclusivo reduziu as concentrações de cobre no soro dos ovinos, bem como a interação cobre-enxofre-molibdênio com as fontes inorgânicas dos minerais reduziu a ceruloplasmina e cobre no soro. Em contrapartida, a ceruloplasmina mostrou aumento nos níveis quando da administração de enxofre orgânico. Os teores de enxofre sanguíneos não foram influenciados pelos tratamentos. No líquido biliar contatou-se efeito do molibdênio exclusivo sobre os minerais, mostrando redução nas concentrações de cobre e enxofre e aumento na concentração de molibdênio. Os valores hepáticos de cobre e enxofre não foram influenciados (P>0,05) pelos tratamentos, somente o molibdênio hepático mostrou efeito na interação cobre-enxofre-molibdênio. A biodisponibilidade do cobre proteinado (145,09%) foi superior à do sulfato de cobre (100%), independente da fonte de enxofre e na ausência e molibdênio, quando determinado pela regressão múltipla \"slope ratio\". / This research aim to study the effect of organic and inorganic copper and sulfur sources in the interaction copper-sulfur-molybdenum, estimating the bioavailability of two sources of copper in the diet of sheep. For that used 40 weaned sheep at about 3 months of age and weighing 20 kg and distributed in 10 treatments, as follows: 1) basal diet; 2) diet containing molybdenum; 3) basal diet + copper inorganic + sulfur inorganic; 4) basal diet + copper inorganic + sulfur organic; 5) basal diet + copper organic + sulfur, inorganic; 6) basal diet + copper organic + sulfur organic; 7) diet with molybdenum + copper inorganic + sulfur inorganic; 8) diet with molybdenum + copper inorganic + organic sulfur; 9) diet with molybdenum + copper organic + sulfur inorganic, 10) diet with molybdenum + organic copper + organic sulfur. The experimental period lasted 84 days, animal weighing was performed on days 0, 28, 56 and 84 days to monitor its development. Blood collections to study levels of copper, sulfur and molybdenum, concentrations of glucose, albumin, urea, cholesterol, triglycerides, hematocrit and ceruloplasmin were also performed. Liver biopsies were made on zero day for copper, sulfur and molybdenum analysis. At the end of the experimental period, the animals were slaughtered and samples were collected from liver and bile fluid for final determination of mineral, and was measured pH and weight of hot and cold carcasses. At the end of the third experimental period was done a metabolic balance of copper, molybdenum and sulfur. The bioavailability of copper was calculated by \"slope ratio\" technique, using the concentration of copper in the liver. The parameters were analyzed considering the existence of a factorial structure 2 x 2 x 2 (with and without molybdenum, organic and inorganic copper and organic and inorganic sulphur) and a basal diet and a basal diet plus molybdenum. The concentrations of glucose, albumin, urea, cholesterol and blood hematocrit levels were not affected (P>0,05) by the addition of molybdenum or adding copper and sulfur, or the copper-molybdenum-sulfur interaction. However, blood triglyceride concentrations were changed (P<0.05) by copper-sulfur interaction, with a reduction in their concentration after the addition of copper and sulfur in organic sources. The live weight of the sheep was not affected (P>0,05) by the treatments, but the weight gain was influenced by the interaction copper-molybdenum-sulfur there was a reduction in weight gain with the addition of molybdenum and an increase was observed in the weight gain when added with copper and molybdenum in the organic sulfur sources compared to the inorganic sources. The supplemental sources of copper and sulfur and addition of molybdenum had no effect (P>0,05) on pH values and carcass weight. The presence of molybdenum in the diet reduced the levels of ceruloplasmin and the exclusive molybdenum presence reduced copper concentrations in the serum of the sheep. The copper-molybdenum-sulfur interaction with the inorganic sources of minerals decreased serum ceruloplasmin and copper. On the other hand, there was an increase in ceruloplasmin levels with organic sulfur administration. The blood levels of sulfur were not affected by treatments. Bile liquid was affected by molybdenum, it was a reduction in the concentrations of copper and sulfur with increase in the molybdenum concentration. The copper and sulfur liver levels were not influenced (P>0,05) by treatments, only the liver Molybdenum effect showed on copper-molybdenum-sulfur interaction. The bioavailability of organic copper (145.09%) was higher than that of copper sulphate (100%), irrespective of the source and in the absence and sulfur and molybdenum, as determined by multiple regression slope ratio.
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Molecular genetic analysis of ceruloplasmin in oesophageal cancerStrickland, Natalie 03 1900 (has links)
Thesis (MSc (Genetics))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: Oesophageal cancer (OC) is characterised by the development of malignant tumours in the epithelial cells lining the oesophagus. It demonstrates marked ethnic variation, with squamous cell carcinoma (SCC) being more prevalent in the Black population and adenocarcinoma (ADC) occurring more often in Caucasians. The aetiology of this complex disease has been attributed to a variety of factors, including an excess of iron (resulting in increased tumourigenesis), oesophageal injury and inflammation.
The present study attempted to determine the mutation spectrum of the regulatory and coding regions of the ceruloplasmin (CP) gene, involved in iron metabolism, in the Black South African OC population. The patient cohort was comprised of 96 (48 male and 48 female) unrelated individuals presenting with SCC of the oesophagus. The control group consisted of 88 unrelated, healthy population-matched control individuals. The techniques employed for mutation detection in this study included polymerase chain reaction (PCR) amplification, heteroduplex single-strand conformation polymorphism (HEX-SSCP) analysis, restriction fragment length polymorphism (RFLP) analysis followed by bidirectional semi-automated DNA sequencing analysis to verify the variants identified.
Mutation detection of CP resulted in the identification of fourteen previously described (5’UTR-567C→G, 5’UTR-563T→C, 5’UTR-439C→T, 5’UTR-364delT, 5’UTR-354T→C, 5’UTR-350C→T, 5’UTR-282A→G, V223, Y425, R367C, D544E, IVS4-14C→T, IVS7+9T→C and IVS15-12T→C) and four novel (5’UTR-308G→A, T83, V246A and G633) variants. Statistical analysis revealed that two of the novel variants were significantly associated with OC in this study; the promoter variant 5’UTR-308G→A (P=0.012) and the exonic variant G633 (P=0.0003). It is possible that these variants may contribute to OC susceptibility in the Black South African population. OC symptoms generally present late in the development of the disease, and as a result treatment after diagnosis is highly ineffective. Early detection of symptoms and subsequent treatment is therefore the most effective manner of disease intervention. In high incidence areas, such as the Transkei region of South Africa, the implementation of a screening programme would be the ideal way to achieve this goal. The information that can be gathered from the identification of potential modifier genes for OC can lead to improvements in early detection, which in turn may lead to advancements in the treatment and counselling to individuals with OC. To our knowledge, this is the first study concerning CP and its effects on iron dysregulation in the Black South African population with oesophageal cancer. / AFRIKAANSE OPSOMMING: Oesofageale kanker word gekenmerk deur die ontwikkeling van kwaardaardige gewasse in die epiteelweefsel van die oesofageale voering. Hierdie siekte demonstreer opvallende etniese variasie, met plaveisel selkarsinoom meer algemeen in die Swart populasie en adenokarsinoom meer algemeen in die Kaukasiese populasie. Die ontwikkeling van hierdie komplekse siekte word aan ‘n aantal faktore toegeskryf, insluitend ‘n oormaat yster (wat lei tot ‘n vermeerdering van gewasse) en oesofageale besering en -ontsteking.
Die doel van die hierdie studie was om die mutasie spektrum van die regulatoriese- en koderingsarea van die ceruloplasmin (CP) geen, betrokke in yster metabolisme, in die Swart Suid Afrikaanse oesofageale kanker populasie te bepaal. Die pasiënt groep het bestaan uit 96 (48 manlik en 48 vroulik) onverwante individue met plaveisel selkarsinoom van die oesofagus. Die kontrole groep het uit 88 nie-geaffekteerde onverwante, populasie spesifieke individue bestaan. Die tegnieke aangewend vir mutasie deteksie in hierdie studie sluit in polimerase kettingsreaksie amplifikasie, heterodupleks enkelstring konformasie polimorfisme analise en restriksie fragment lengte polimorfisme analise, gevolg deur tweerigting semi-geoutomatiseerde DNS volgorde-bepalingsanalise om die geïdentifiseerde variante te bevestig.
Mutasie deteksie van CP het tot die identifikasie van veertien reeds beskryfde (5’UTR-567C→G, 5’UTR-563T→C, 5’UTR-439C→T, 5’UTR-364delT, 5’UTR-354T→C, 5’UTR-350C→T, 5’UTR-282A→G, V223, Y425, R367C, D544E, IVS4-14C→T, IVS7+9T→C en IVS15-12T→C) en vier nuwe (5’UTR-308G→A, T83, V246A en G633) variante gelei. Statistiese analise het getoon dat twee van die nuwe variante betekenisvol geassosieerd was met oesofageale kanker in hierdie studie; die promotor variant 5’UTR-308G→A (P=0.012) en die eksoniese variant G633 (P=0.0003). Dit is moontlik dat hierdie variante mag bydra tot oesofageale kanker vatbaarheid in die Swart Suid Afrikaanse populasie.
Oesofageale kanker simptome vertoon gewoonlik op ‘n latere stadium in die ontwikkelingsproses van die siekte, en as ‘n gevolg is behandeling na diagnose hoogs oneffektief. Vroegtydige identifikasie van die simptome en daaropvolgende behandeling is die mees effektiewe manier vir ingryping. In hoë voorkoms streke, soos die Transkei gebied van Suid Afrika, sal die implementasie van ‘n siftingsprogram die ideale manier wees om hierdie doel te bereik. Die inligting wat dan versamel word, insluitend identifisering van modifiserende gene vir oesofageale kanker, kan lei tot ‘n verbetering in vroegtydige deteksie van die siekte. In effek kan dit dan lei tot beter behandeling en berading vir individue met oesofageale kanker. So ver ons kennis strek, is hierdie die eerste studie wat CP en sy effek op yster disregulasie in die Swart Suid-Afrikaanse populasie met oesofageale kanker behels.
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Avaliação de fontes de cobre para ovinos com ensaio de biodisponibilidade / Influence of diferente levels and sources of copper supplementation with bioavailability studyCarolina Yumi Cascão Yoshikawa 21 March 2014 (has links)
O objetivo do estudo foi estimar a biodisponibilidade de duas fontes de cobre: orgânica (cobre metionina) e inorgânica (sulfato de cobre) na dieta de cordeiros. O experimento foi conduzindo na FZEA USP de Pirassununga utilizando 40 cordeiros da raça Merino X Texel, que foram distribuídos aleatoriamente em cinco grupos, e submetidos a cinco tratamentos, totalizando oito animais por tratamento: Tratamento 0: Dieta controle sem adição de Cu; Tratamento 1: 10 mg de Cu/Kg de MS na forma de sulfato de Cu; Tratamento 2: 30g de Cu/Kg de MS na forma de sulfato de Cu; Tratamento 3: 10 mg de cu/kg de MS na forma de cobre metionina; Tratamento 4: 30 mg de cu/kg de MS na forma de cobre metionina. Foram feitas biópsias do fígado dos animais no tempo zero para análise de cobre e colhidas amostras de sangue nos dias 0, 28, 56 e 84 dias para determinação de Cu sérico, atividade de ceruloplasmina e enzimas de função hepática. Ao final do experimento, os animais foram abatidos para colheita de amostras de fígado, músculo e rim, para determinação dos teores de Cu e da enzima superóxido dismutase (SOD). Nos últimos dez dias do experimento foi realizado um balanço metabólico de cobre. A biodisponibilidade foi calculada pela técnica \"slope ratio\", utilizando como parâmetros a concentração de cobre no fígado. Não houve diferença (P>0,05) no desempenho dos animais (peso vivo e ganho de peso) entre os tratamentos. A concentração sérica de AST e ALT permaneceu abaixo dos níveis de intoxicação em todos os tratamentos, durante todo o período. A atividade da ceruloplasmina não diferiu entre os tratamentos (P>0,05). O teor de cobre no soro, na biópsia do fígado e no músculo não foi diferente (P>0,05) entre os tratamentos. Entretanto, a concentração do mineral no fígado dos animais suplementados (284,28 mg/kg) foi maior (P<0,05), quando comparados ao grupo controle (168,01 mg/kg), assim como o Cu-met 30 mg/kg (341,29 mg/kg) foi superior (P<0,05) ao de 10mg/kg MS (263,02 mg/kg). A atividade da SOD nos animais suplementados (µmol/mg prot foi superior à do grupo controle. Nos rins o teor de cobre foi superior nos animais que receberam 30mg/kg de MS de Cu-met (6,65 mg/kg) em relação aos que receberam 10 mg/kg de MS da mesma fonte (3,86 mg/kg). A absorção e a retenção aparentes do cobre foram maiores para a fonte inorgânica, comparada com a orgânica. A biodisponibilidade do cobre determinada pela concentração de cobre no fígado, utilizando a técnica do \"slope ratio\", considerando o CuSO4 como padrão (100%), apresentou disponibilidade de 150,64% para o Cu-met. / The study was conducted to estimate the relative bioavailability of two sources of supplemental copper: organic (copper methionine) and inorganic (copper sulfate) in the diet of lambs, by analyzing the concentration of copper and enzymes in the liver and metabolic balance calculation, using 40 lambs breed Merino X Texel, which was fed three concentrations of copper (basal + two additions) in two sources, which were randomly allotted to five groups, and subjected to five treatments: treatment 0: control (diet without addition of Cu); treatment 1: (diet with 10 mg Cu/Kg DM of CuSO4); Treatment 2: (diet with 30 g Cu/Kg DM of CuSO4); Treatment 3: (diet with 10 mg Cu/kg DM of copper methionine; Treatment 4: (diet with 30 mg cu/kg DM of copper methionine). Liver biopsies were made on 0 d. Blood samples were taken via the jugular vein on 0, 28, 56 and 84 d to determine serum Cu and serum ceruloplasmin and liver transaminases (AST, ALT) concentrations. The animals were slaughtered and samples of liver, kidney and muscle were taken for the determination of the levels of Cu and superoxide dismutase activity. In the last ten days of the experiment a metabolic balance of copper was conducted. The bioavailability was calculated by the \"slope ratio\" technique, using the concentration of copper in the liver as parameter. There was no difference (P >0.05) on animal performance (live weight and weight gain) among treatments. The serum AST and ALT levels remained below poisoning in all treatments during the period. The ceruloplasmin activity did not differ between treatments (P>0.05). The copper content in biopsy, serum and muscle was not different (P>0.05) between treatments. However, the mineral concentration in the liver of animals fed (284.28 mg/kg) was higher (P <0.05) when compared to the control group (168.01 mg/kg ) and 30 Cu -met mg/kg (341.29 mg/kg) was higher (P <0.05) at 10mg/kg MS (263.02 mg/kg). The SOD activity in the supplemented animals (mmol/mg prot) was superior to the control group. Copper in Kidneys was higher in animals that received 30mg/kg MS meth-Cu (6.65mg/kg) compared those receiving 10 mg/kg DM from the same source (3.86 mg/kg). Apparent absorption and retention of copper were higher for inorganic source, compared with the organic. The bioavailability determined by the concentration of copper in the copper liver, using the technique of \"slope ratio\", considering CuSO4 as standard ( 100% ) presented availability of 150.64 % for Cu-met.
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