Orientador: Ana Silvia Alves Meira Tavares Moura / Banca: Simone Fernandes / Banca: Valderez Bastos Valero-Lapckick / Banca: Denose Rangel da Silva Sartori / Banca: Luiz Edivaldo Pezzato / Resumo: O objetivo deste estudo foi avaliar a preferência e efeito do fornecimento de materiais de nidificação sobre o desempenho reprodutivo de camundongos isogênicos da linhagem BALB/c e da linhagem heterogênica Swiss em sistema de acasalamento intensivo monogâmico. Primeiro foi realizado um estudo para avaliar a preferência dos camundongos pelo material oferecido para nidificação. Utilizou-se um sistema composto de quatro gaiolas, com livre acesso a água e ração, interligados por tubos de PVC que permitiam que os animais se locomovessem entre todas as gaiolas. Quatro tipos de materiais foram oferecidos para a construção do ninho: algodão, gaze, rolinho de papelão, e touca de polipropileno descartável. Cada um dos quatro materiais foi oferecido simultaneamente em uma das quatro gaiolas que compunham o sistema. Foram usados 10 sistemas iguais e cada um abrigou um casal da linhagem BALB/c, desde os 28 dias de idade até o terceiro ciclo reprodutivo. Os mais encontrados na confecção do ninho, em ordem decrescente, foram a touca, rolinho, algodão e gaze (P< 0,0083). Com base nestes resultados foram selecionados dois tipos de materiais para fornecimento aos animais no experimento 2. Embora o rolinho tenha sido o segundo material mais utilizado optou-se pelo algodão devido a inviabilidade de fornecimento do item durante todo o período de duração do experimento 2. O segundo experimento avaliou a eficiência reprodutiva em cinco ciclos reprodutivos do nascimento até o desmame. Usou-se 60 casais de irmãos completos da linhagem BALB/c (isogênica) e 60 casais formados por acasalamentos aleatórios da linhagem Swiss (heterogênica) de padrão sanitário controlado, criados e mantidos em ambiente padronizado. Eles foram distribuídos, num delineamento inteiramente casualizado, em arranjo fatorial 2x2 (duas linhagens em alojamento com ou sem material para nidificação). Como forma de ... / Abstract: The objective of this study was to evaluate the preference and effect of nesting materials provision on performance of inbred mice of the BALB/c strain and of heterogenic Swiss in intensive monogamous mating system. Firstly, a study was conducted to evaluate the preference of mice for the material offered for nesting. A system composed of four cages was used, with free access to water and food, connected by PVC tubing, which allowed animal displacement among all cages. Four types of materials were available for construction of the nest: cotton, gauze, cardboard rolls, and disposable polypropylene cap. Each of the four materials was offered simultaneously in one of four cages that formed the system. Ten identical system were used and each one housed a BALB/c couple, from 28 days of age up to the third reproductive cycle. The most commonly found materials in nest making were cap, roll, cotton and gauze (P <0.0083). Based on these results, two types of materials were selected to be offered to the animals in experiment 2. Although the roll was the second most used material, we chose cotton due to unfeasibility supplying of the item throughout the duration of the second experiment. The second experiment evaluated the reproductive efficiency in five reproductive cycles from birth to weaning. Sixty BALB / c full siblings pairs (inbred) and 60 pairs formed by random mating of Swiss strain (outbred), bred and maintained in a standardized environment were used. They were distributed in a completely randomized design in a 2x2 factorial arrangement (two strains in housing with or without nesting material). As a way of cage enrichment, polypropylene disposable cap cut into 8 pieces of approximately 1 cm each and one piece of cotton about 3 g were used after being previously packaged and autoclaved . The following characteristics were evaluated: age at first parturition, litter intervals, pre-weaning mortality and ... / Doutor

From locating a secure home, foraging for food, running away from predators, spatial navigation is an integral part of everyday life. Multiple brain regions work together to form a three-dimensional representation of our environment; specifically, place cells, grid cells, border cells & head direction cells are thought to interact and influence one another to form this cognitive map. Head direction (HD) cells fire as the animal moves through space, according to directional orientation of the animal’s head with respect to the laboratory reference frame, and are therefore considered to represent the directional sense. Interestingly, inactivation of head direction cell-containing brain regions has mixed consequences on spatial behavior. Current methods of identifying HD cells are limited to in vivo electrophysiological recordings in a dry-land environment. We first developed a dry-land version of the MWM in order to carry out behavioral-recording paired studies. Additionally, to learn about HD cells function we quantified expression of neuronal activation marker (c-Fos), and L-amino acid transporter 4 (Lat4) in neurons found within the HD cell dense anterodorsal thalamic nucleus (ADN) in mice after exploratory behavior in an open field, or forward unidirectional movement on a treadmill. We hypothesize that the degree to which ADN neurons are activated during exploratory behavior is influenced by the range of heading directions sampled. Additionally, we hypothesize that c-Fos and Lat4 are colocalized within ADN neurons following varying amounts of head direction exposure. Results indicate that following free locomotion of mice in an open field arena, which permitted access to 360° of heading, a greater number of ADN neurons express c-Fos protein compared to those exposed to a limited range of head directions during locomotion in a treadmill. These findings suggest that the degree of ADN neuronal activation was dependent upon the range of head directions sampled. We observed a high degree of colocalization of c-Fos and Lat4 within ADN suggesting that Lat4 may be a useful tool to manipulate neuronal activity of HD cells. Identifying genetic markers specific to ADN helps provide an essential understanding of the spatial navigation system, and supports development of therapies for cognitive disorders affecting navigation. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2017. / FAU Electronic Theses and Dissertations Collection

Preliminary analyses of three kinds of promoter activity showed that a-actin gene promoter was chosen to initiate the hormone transcription for the first consideration. We have fused the cDNAs encoding the intact somatolactins in frame to a zebrafish a-actin gene promoter to generate transgenic zebrafish lines co-injected with a GFP protein driven by the same promoter. The transgenic zebrafish were selected from GFP expression and confirmed by genomic PCR and Southern blot analysis, then maintained as transgenic founders. Measurement of the transgenes' expressions and the expressions of marker genes in different pathways by using real-time PCR provided a general understanding of SLs' actions. The data obtained indicated that the over-expressing of SLalpha and SLbeta in vivo significantly enhance the transcriptions of the insulin-like growth factors, IGF1 (5.46-fold and 6.77-fold), IGF2a (4.38-fold and 4.35-fold) and IGF2b (2.83-fold and 3.94-fold), but down-regulated IGF3 (a novel member found specifically in gonad) in larvae. However, the stimulation by administration of recombinant proteins (SLalpha and SLbeta) only showed a slight induction of the mRNA levels of IGFs (IGF1, IGF2a and IGF2b) on ZFL cells in vitro. / Somatolactin (SL) is a novel member of pituitary polypeptide hormone found only in fish; it shares significant structural homology with prolactin and growth hormone. Since somatolactin receptor (SLR) was first defined as GHR1 and orthologous to the growth hormone receptor GHR2, SL and GH may share similar actions in growth and development. Recently, two SLs have been identified as SLalpha and SLbeta with similar structures, freshwater fish have these two isoforms found in the same species and only one isoform (SLalpha) is found in marine species. The two isoforms of SL may have different functions and physiological actions. To investigate the roles of SLs on vertebrate development and embryogenesis, we generated transgenic fish models with "all zebrafish" elements in origin to study the physiological functions of SLs in zebrafish. / The ectopic expression of somatolactins also results in up-regulating gene expression of insulin, leptin, sterol regulatory element binding protein 1 (SREBP1) and fatty acid synthase (FAS), as well as the expression of vitellogenin and melanocyte-stimulating hormone (MSH) levels while causing reduction of catalase (CAT) and glutathione S-transferase (GST) levels in larvae. The results here represent the similar function between SLalpha and SLbeta and reveal more details in fish of the endocrinology system involvement in growth development, glucose synthesis, lipid metabolism, reproduction, pigmentation and antioxidant defense system through the actions of SLs. / Three different gene promoters of zebrafish have been isolated to initiate the ectopic expression of somatolactins in vivo, which including a constitutional beta-actin gene promoter, a liver specific transferrin gene promoter and a zinc ion inducible metallothionein (MT) gene promoter. The promoter activities were tested in fish cell-line by using luciferase reporter assay. In MT gene promoter, two alleles of a zebrafish metallothionein II gene (zMT-II) promoter (zMT-IIA and zMT-IIB) containing 10 MREs in the 5'-flanking region (1,514 bp) were identified in zebrafish. These putative MREs were confirmed via electrophoretic mobility shift assay (EMSA) to have binding activities from the cellular and nuclear extracts of a zebrafish cell line, ZFL. Transient gene expression studies using zebrafish liver (ZFL) cell lines also confirmed that the most distal cluster of MREs contributed to the maximal induction of zMT-IIA activity by Zn2+ and the Zn 2+ induction was dose-dependent. EMSA also identified transcription factor(s) of two different sizes from the cytoplasmic and nuclear extracts of the ZFL cells that were able to bind with the MREs, but no increase in MRE binding was detected in the extracts of these cells after Zn2+ or Cd2+ treatment, compared with untreated control cells. The mechanisms of MT gene transcription induction via metal ions are discussed herein. / Wan, Guohui. / Adviser: Chan King Ming. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 139-163). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Data of histochemical studies suggest that THs exert significant influences on myelination and the process of neuron degenerations leading to deficit in brain development. Results of IHC in this study are not fully matched to gene transcription results. It indicates that gene transcriptions may not be synonymous with gene translation. The ratio of RNA transcripts to proteins may differ among genes. It suggested that the transcriptional and morphological studies are supplementary to each other. / In this study, hypothyroidism was induced by different regimens. They are 0.05% methimazole (MMI), 0.02% MMI plus 1% NaC1O4, and 0.1% propyl-thiouracil in drinking water of mother rats from day 16 of their pregnancies, postnatal day 1 or day 4 to day 24. The replacement therapy was done by giving either a single s.c. injection of 300 nM T4 18 hrs before sacrifice on postnatal day 16, or by giving s.c. daily injections of 1.5 ng T3 plus 9 ng T4 per gram body weight from postnatal day 11 to 15. Olfactory bulb (OB), hippocampus (H) and cerebellum (CM) and in some cases also cerebral cortex (CX) were studied. The axon guidance molecules and their related genes, Galpha proteins, RGSs and small GTPases mRNA levels were examined with Real-Time quantitative RT-PCR. Special staining on sagittal frozen brain sections with histochemical and immunohistochemical (IHC) techniques were also studied. / It is well established that neonatal hypothyroidism causes defective development of the brain. As signals for synaptogenesis, growth factors and their receptors regulate the gene expression of the growth cone proteins and axon guidance molecules, and control the differentiation of neurons during brain development. Galpha proteins are signal transducers and regulators of G protein signaling (RGS) proteins are recently identified family of proteins that dampen G protein-coupled receptor-mediated signaling by accelerating the intrinsic GTPase activity of Galpha proteins. They play important roles in determining the intensity and specificity of signaling pathways in brain and their adaptations to synaptic activities. / The transcript abundance of some genes, such as Galphai1, Galphai2, Galphaol, Galphas, RGS 2, - 4, -5, -7, -8, -12, -16, -17, Mfn2, TRalpha1, TRbeta 1, Rheb, Rhes, Dexras1, Plexin1, Citron-Kinase, GAP-43, CRMP1, CRMP3, CRMP4 and CRMP5 in CM; Galpha12, Galphai3, G alphaol, Galphaolf, Galphaq, RGS 5, - 7, -8, -16, cdc42, Rhes, TC10, Dexras1, Citron-Kinase, TRalpha1, CRMP2, Wnt7A, Sema3A and GAP43 in OB and Galpha12, Galphai1 , Galphai2, Galphao2, RGS 5, -8, - 16, Mfn1, Mfn2, TRalpha1, Rhes, TC10, Dexras1, Citron-Kinase, GAP-43, CRMPI1, CRMP4, CRMP5 and Gda in H; TRalpha1, CRMP1, CRMP4, CRMP5, Plexin1, Plexin2, Gda and GAP-43 in CX are significantly altered in the neonatal hypothyroid rats. Of note, the mRNA levels of several genes were normalized by TH replacement therapy. Close correlations were found among various Galpha proteins, RGS genes, small GTPases and some axon guidance molecules in a brain- region- specific manner. Our results indicate certain direct or indirect transcriptional effects of the THs on the expression of brain development-related genes and these effects are probably under both temporal and spatial regulations during brain development. / by Yan, Ran. / "January 2007." / Advisers: Michael S. C. Tam; Chun Cheung Wong. / Source: Dissertation Abstracts International, Volume: 68-09, Section: B, page: 5776. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 199-238). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

高膽固醇血症是產生動脈粥樣硬化的危險因素，本研究旨在探討甾醇的氧化產物和植物化學物質在餵食高膽固醇食金黃地鼠模型中對脂蛋白代謝的影響及其相關機制。 / 本研究包含四個部分。食物中同時含有植物甾醇及其氧化產物。第一部分旨在研究β-穀甾醇（Si）、甾醇(St)、β-穀甾醇氧化產物(SiOP)和豆甾醇氧化產物(StOP)對金黃地鼠血脂的影響。本研究顯示，Si和St組能有效降低血總膽固醇（TC）、非高密度脂蛋白膽固醇（non-HDL-C）和甘油三酯（TAG）的水平，而SiOP和StOP則失去此能力。RT-PCR分析表明，Si和St而非SiOP和StOP，能下調腸道醯基輔酶A：膽固醇醯基轉移酶2（ACAT2）和微粒體甘油三酯轉移蛋白（MTP）的mRNA表達。Si和St而非SiOP和StOP能有效防止動脈粥樣硬，Si和St的動脈弓舒張能力強於對照組和SiOP、StOP組。 / 辣椒鹼是辣椒中的活性成分。本研究第二部分表明，辣椒鹼能降低TC，NON-HDL-C，TAG，而不影響高密度脂蛋白膽固醇。餵養辣椒鹼能增加糞便中總酸性固醇的排泄，此作用有可能是通過上調膽固醇7α-羥化酶（CYP7A1）和下調肝X受體α（LXRα）的基因表達來實驗。辣椒鹼可通過抑制COX-2基因表達來改善內皮依賴性收縮。 / 藍莓含有豐富的抗炎抗氧化劑，例如花青素。本研究第三部分表明，食物中添加0.5和1.0藍莓花青素能導致TC呈劑量效益地降低6-12%，其中還伴隨22-29的中性固醇和41-74%的膽汁酸排泄的增加。RT-PCR分析表明食物中添加的藍莓花青素能下調腸道Niemann-Pick C1 Like 1 (NPC1L1)，ACAT2，MTP， 腺苷三磷酸結合盒轉運體G8(ABCG8)和肝臟3-羥基-3-甲基戊二醯輔酶A還原酶(HMG-CoA　Reductase)的基因表達。 / 芝麻素是芝麻種子中含有抗氧化活性的木脂素類化合物。本研究第四部分表明，在食物中添加芝麻素可有效調控TC和non-HDL-C，同時不影響TAG，並導致非高密度脂蛋白膽固醇與高密度脂蛋白膽固醇比例的下降。這有可能與膽汁酸排泄增加、CYP7A1基因的上調，LXR的下調有關。 / 綜上所述，本研究證實了植物甾醇、辣椒鹼、藍莓花青素和芝麻素降低血膽固醇的能力。與此同時，本研究還表明植物甾醇被氧化後將失去其降低膽固醇的能力。 / Hypercholesterolemia is a major risk factor in the development of atherosclerosis. Functional foods that can lower or regulate cholesterol concentration are of interest to both public and scientific communities. The present study was to investigate the effects of phytosterols, phytosterol oxidation products (POPs), capsaicinoids, blueberry anthocyanins and sesamin on plasma cholesterol concentration using hamsters as a model. / The whole project consisted of four parts. Human diets contain both phytosterols and POPs. Part I was to examine the effect of β-sitosterol (Si), stigmasterol (St), β-sitosterol oxidation products (SiOP) and stigmasterol oxidation products (StOP) on plasma cholesterol concentration. Results showed both Si and St could reduce while SiOP and StOP lost the capacity of lowering plasma total cholesterol (TC), non-high density lipoprotein cholesterol (non-HDL-C) and triacylglycerols (TAG). Real-Time PCR analysis demonstrated Si and St but not SiOP and StOP down-regulated mRNA levels of intestinal acyl CoA: cholesterol acyltransferase 2 (ACAT2) and microsomal triglyceride protein (MTP). In addition, aortas from hamsters given diets containing Si and St relaxed better than those from the control and their corresponding SiOP- and StOP-treated hamsters, suggesting that Si and St not SiOP and StOP were beneficial in improving lipoprotein profile and aortic function. / Capsaicinoids refer to a group of pungent compounds that are the active components found in chili peppers. Part II was to investigate the cholesterol-lowering activity of capsaicinoids and the associated molecular mechanisms. Results demonstrated that capsaicinoids reduced plasma TC, non-HDL-C and TAG with high-density lipoprotein cholesterol (HDL-C) being unaffected. This was accompanied by an increase in the fecal excretion of total acidic sterols, possibly mediated by up-regulation of cholesterol 7α-hydroxylase (CYP7A1) and down-regulation of liver X receptor alpha (LXRα). Capsaicinoids could also improve the endothelium-dependent relaxations and reduce the endothelium-dependent contractions by inhibiting the gene expression of COX-2. / Blueberries are rich in anthocyanins. Results from Part III experiments demonstrated that dietary supplementation with 0.5 and 1.0 % blueberry anthocyanins for 6 weeks decreased plasma TC concentration by 6-12% in a dose-dependent manner. This was accompanied by increasing the excretion of fecal neutral and acidic sterols by 2229% and 4174%, respectively. Real-time PCR analyses demonstrated that incorporation of blueberry anthocyanins into diet down-regulated the genes of intestinal Niemann-Pick C1-like 1 (NPC1L1), ACAT2, MTP, ABCG 8 and hepatic 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase. / Sesamin is a major lignan in sesame seed and is known to exhibit antioxidative activity. Part IV was to investigate the mechanism by which sesamin decreased plasma cholesterol concentration. Results clearly demonstrated supplementation of sesamin into diets could favorably reduce serum TC and non-HDL-C with TAG being unaffected. In addition, dietary supplementation of 0.2 or 0.5% of sesamin could cause a significant decrease in the ratio of non-HDL-C to HDL-C. This was accompanied by a marked increase in bile acid excretion and up-regulation of CYP7A1 and down-regulation of LXRα. / In conclusion, phytosterols, capsaicinoids, blueberry anthocyanins and sesamin were beneficial in improving lipoprotein profile in hamsters fed a high-cholesterol diet. However, phytosterols lose the cholesterol-lowering capacity when they are oxidized. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Liang, Yintong. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 112-123). / Abstracts also in Chinese. / Chapter Chapter 1 --- General Introduction / Chapter 1.1 --- Cardiovascular diseases --- p.1 / Chapter 1.2 --- Cholesterol --- p.2 / Chapter 1.3 --- Lipoproteins --- p.4 / Chapter 1.4 --- Cholesterol homeostasis --- p.6 / Chapter 1.4.1 --- HMG-CoA reductase --- p.7 / Chapter 1.4.2 --- LDL receptor --- p.9 / Chapter 1.4.3 --- Intestine ACAT2 --- p.10 / Chapter 1.4.4 --- NPC1L1 --- p.11 / Chapter 1.4.5 --- CYP7A1 and LXRα --- p.12 / Chapter 1.4.6 --- SREBP2 --- p.14 / Chapter 1.4.7 --- ABCG5 and ABCG8 --- p.15 / Chapter 1.5 --- Phytochemicals --- p.16 / Chapter 1.5.1 --- Phytosterols --- p.16 / Chapter 1.5.2 --- Capsaicinoids --- p.17 / Chapter 1.5.3 --- Blueberry anthocyanins --- p.19 / Chapter 1.5.4 --- Sesamin --- p.20 / Chapter 1.6 --- Animal model --- p.22 / Chapter Chapter 2 --- Effect of Phytosterols and their Oxidation Products on Lipoprotein Profiles and Vascular Function / Chapter 2.1 --- Introduction --- p.23 / Chapter 2.2 --- Objective --- p.24 / Chapter 2.3 --- Materials and methods --- p.24 / Chapter 2.3.1 --- Preparation of sitosterol oxidation products (SiOP) and stigmasterol oxidation products (StOP) / Chapter 2.3.2 --- Diets --- p.25 / Chapter 2.3.3 --- Hamsters --- p.25 / Chapter 2.3.4 --- Analysis of individual SiOP and StOP in serum and liver --- p.26 / Chapter 2.3.5 --- Analysis of plasma lipoproteins --- p.28 / Chapter 2.3.6 --- Measurement of atherosclerotic plaque --- p.28 / Chapter 2.3.7 --- Analysis of cholesterol in the liver and aorta --- p.28 / Chapter 2.3.8 --- Determination of fecal neutral and acidic sterols --- p.29 / Chapter 2.3.9 --- Real-time PCR analysis of mRNA of liver SREBP2, LDL receptor, HMG-CoA reductase, CYP7A1, LXRα, and small intestine NPC1L1, ABCG5, ABCG8, ACAT2, MTP. --- p.29 / Chapter 2.3.10 --- Western blotting analysis of hepatic SREBP2, LDL receptor, HMG-CoA reductase, LXRα and CYP7A1 --- p.32 / Chapter 2.3.11 --- Vascular reactivity --- p.32 / Chapter 2.4 --- Results --- p.34 / Chapter 2.4.1 --- Composition of SiOP and StOP --- p.34 / Chapter 2.4.2 --- Food intake, body and organ weights --- p.34 / Chapter 2.4.3 --- Plasma TC, HDL, non-HDL ,TAG, Non-HDL-C/HDL-C --- p.34 / Chapter 2.4.4 --- Aortic cholesterol and atherosclerotic plaque --- p.35 / Chapter 2.4.5 --- Liver cholesterol, SiOP and StOP --- p.35 / Chapter 2.4.6 --- Fecal neutral, acidic sterols and cholesterol balance --- p.35 / Chapter 2.4.7 --- Immunoblot and mRNA analysis --- p.36 / Chapter 2.4.8 --- Vascular reactivity --- p.36 / Chapter 2.4.9 --- Role of COX in endothelium-dependent contractions --- p.37 / Chapter 2.5 --- Discussion --- p.50 / Chapter Chapter 3 --- Cholesterol-Lowering Activity of Capsaicinoids Is Mediated by Increasing Sterol Excretion in Hamsters Fed a High Cholesterol Diet / Chapter 3.1 --- Introduction --- p.54 / Chapter 3.2 --- Objective --- p.55 / Chapter 3.3 --- Materials and methods --- p.55 / Chapter 3.3.1 --- Diets --- p.55 / Chapter 3.3.2 --- Hamsters --- p.57 / Chapter 3.3.3 --- Analysis of plasma lipoproteins --- p.57 / Chapter 3.3.4 --- Measurement of atherosclerotic plaque --- p.57 / Chapter 3.3.5 --- Analysis of cholesterol in the liver and aorta --- p.57 / Chapter 3.3.6 --- Determination of fecal neutral and acidic sterols --- p.57 / Chapter 3.3.7 --- Real-time PCR analysis of mRNA of liver SREBP2, LDL receptor, HMG-CoA reductase, CYP7A1, LXRα, and small intestine NPC1L1, ABCG5, ABCG8, ACAT2, MTP --- p.57 / Chapter 3.3.8 --- Western blotting analysis of hepatic SREBP2, LDL receptor, HMG-CoA reductase, LXRα and CYP7A1 --- p.58 / Chapter 3.3.9 --- Vascular reactivity --- p.58 / Chapter 3.4 --- Results --- p.59 / Chapter 3.4.1 --- Food intake, body and organ weights --- p.59 / Chapter 3.4.2 --- Plasma TC, HDL, non-HDL,TAG, Non-HDL-C/HDL-C --- p.59 / Chapter 3.4.3 --- Aortic cholesterol and atherosclerotic plaque --- p.59 / Chapter 3.4.4 --- Fecal neutral, acidic sterols and cholesterol balance --- p.59 / Chapter 3.4.5 --- Immunoblot and mRNA analysis --- p.60 / Chapter 3.4.6 --- Vascular reactivity --- p.60 / Chapter 3.4.7 --- Role of COX in endothelium-dependent contractions --- p.61 / Chapter 3.5 --- Discussion --- p.74 / Chapter Chapter 4 --- Effect of Blueberry Anthocyanins on Lipoprotein Profiles in Hamsters Fed a Cholesterol Diet / Chapter 4.1 --- Introduction --- p.77 / Chapter 4.2 --- Objective --- p.78 / Chapter 4.3 --- Materials and methods --- p.78 / Chapter 4.3.1 --- HPLC analysis of blueberry anthocyanins --- p.78 / Chapter 4.3.2 --- Diet --- p.79 / Chapter 4.3.3 --- Hamsters --- p.80 / Chapter 4.3.4 --- Analysis of plasma lipoproteins --- p.80 / Chapter 4.3.5 --- Analysis of cholesterol in the liver --- p.80 / Chapter 4.3.6 --- Determination of fecal neutral and acidic sterols --- p.81 / Chapter 4.3.7 --- Real-time PCR analysis of mRNA of liver SREBP2, LDL Receptor, HMG-CoA Reductase, CYP7A1, LXRα, and small intestine NPC1L1, ABCG5, ABCG8, ACAT2, MTP --- p.81 / Chapter 4.3.8 --- Western blotting analysis of hepatic SREBP2, LDL Receptor, HMG-CoA reductase, LXRα and CYP7A1 --- p.81 / Chapter 4.4 --- Results --- p.82 / Chapter 4.4.1 --- Food intake, body, and organ weights --- p.82 / Chapter 4.4.2 --- Plasma TC, HDL-C, non-HDL-C, and TAG --- p.82 / Chapter 4.4.3 --- Liver cholesterol concentration --- p.82 / Chapter 4.4.4 --- Fecal total sterols and apparent sterol retention --- p.82 / Chapter 4.4.5 --- Immunoblot and mRNA analysis --- p.83 / Chapter 4.5 --- Discussion --- p.92 / Chapter Chapter 5 --- Effect of Sesamin on Lipoprotein Profiles in Hamsters Fed a high Cholesterol Diet / Chapter 5.1 --- Introduction --- p.95 / Chapter 5.2 --- Objective --- p.95 / Chapter 5.3 --- Materials and methods --- p.96 / Chapter 5.3.1 --- Diets --- p.96 / Chapter 5.3.2 --- Hamsters --- p.96 / Chapter 5.3.3 --- Methods --- p.97 / Chapter 5.4 --- Results --- p.98 / Chapter 5.4.1 --- Food intake, body and organ weights --- p.98 / Chapter 5.4.2 --- Plasma TC, HDL-C, non-HDL-C ,TAG, Non-HDL-C/HDL-C --- p.98 / Chapter 5.4.3 --- Liver cholesterol --- p.98 / Chapter 5.4.4 --- Fecal neutral, acidic sterols and cholesterol balance --- p.98 / Chapter 5.4.5 --- Immunoblot and mRNA analysis --- p.99 / Chapter 5.5 --- Discussion --- p.109 / References --- p.112

Apesar dos inúmeros grupos de pesquisa de todo o mundo investigando o câncer, esta doença tem se ampliado significativamente. No caso do câncer de ovário, a despeito dos notáveis avanços observados nos tratamentos atuais, um desafio persiste: a incapacidade da preservação de oócitos em tratamento com quimioterapia, radioterapia e/ou cirurgia, nos quais são freqüentes os efeitos colaterais tais como a perda folicular, a infertilidade, a menopausa precoce e as falências ovarianas pós-quimioterapia, decorrentes da irradiação e pós-cirurgia. Neste sentido, o transplante de tecido ovariano pode representar uma alternativa na recuperação da capacidade fértil da mulher, bem como uma terapia de reposição hormonal após tratamento. No presente estudo, foram utilizadas linhagens de camundongos para avaliar a viabilidade do transplante ovariano antes e após a irradiação, como alternativa para a restauração das funções reprodutivas. Para tanto, fêmeas das linhagens isogênicas C57BL/6/Unib e híbridas B6CF1/Unib, foram acasaladas com machos da linhagem C57BL/6/Unib, empregandose diferentes protocolos com fêmeas submetidas ou não aos efeitos da radiação gama de Co-60 nas doses de 4 Gy e 6 Gy. Os resultados demonstram que a transferência ovariana restabeleceu a capacidade reprodutiva dos animais para níveis muito próximos dos normais; que o tamanho médio da ninhada, não revelou diferenças significativas após a transferência ovariana; que os fragmentos do ovário da receptora permanecem funcionais em períodos próximos pós à irradiação com 4 Gy e 6 Gy e que a irradiação nas doses de 4 Gy e 6 Gy destroem as células germinativas comprometendo a reprodução. / Despite numerous research groups, around the world, investigating the cancer, the disease has expanded significantly. In the case of the ovarian cancer, in spite of the notable advances seen in current treatments, a challenge remains: the incapacity to cryopreserve the oocyte in cancer chemotherapy, radiotherapy or surgery, in which side effects are frequent such as follicular loss, infertility, early menopause and ovarian bankruptcy after chemotherapy, after irradiation and post-surgery. In this regard, the transplant of ovarian tissues may be an alternative in recovering the women fertility, as well a hormone replacement therapy after treatment. In our study, we used inbred strains and hybrid mice to assess the viability of ovarian transplantation before and after irradiation, as an alternative for the restoration of the reproductive functions. For that female from the C57BL/6/Unib inbred strain and from the hybrid, B6C-F1/Unib mated with C57BL/6/Unib males, employing different protocols with irradiated and non irradiated females with doses of 4 Gy and 6 Gy of Co60 gamma radiation. The results demonstrate that the ovarian transfer restored the reproductive capacity of the females to almost the normal levels; that the litter size average (production index) showed no significant differences after the ovarian transfer; that the fragments from the ovary of the recipient female are functional in periods coming after irradiation with 4 Gy and 6 Gy and that the irradiation with 4 Gy and 6 Gy destroys the germ cells, compromising the reproduction.

Um condicionador de ar para biotérios foi montado com módulos termoelétricos de efeito Peltier. Para troca térmica, foram testados: 1. dissipação externa a ar, com &delta;t de 14°C, rendimento de 16,46%, consumo de 1212 W/h e, 2. dissipação externa água, com &delta;t de 21°C, rendimento de 46,02%, consumo de 524 W/h. A simulação matemática de operação, com mistura de ar não condicionado, mostrou que o sistema pode servir, na dissipação a ar, a aproximadamente 91 microisoladores padrão rato e a aproximadamente 137, na dissipação a água. Quando comparado com um sistema de compressão de freon, o termoelétrico mostrou economia de 26% na implantação e 38% no consumo elétrico por BTU gerado. O sistema termoelétrico mostrou ainda, precisão de &plusmn; 0,1°C, nas temperaturas experimentais, o que é impossível num sistema de freon. Para os testes em animais foram empregados Ratos wistar, mantidos individualmente, em gaiolas metabólicas de arame, sem abrigo, em sistema microambiental, sob fluxo direto de ar a 0,6 m/s, nas temperaturas de 22°, 24°, 26°, 28° e 30°C (E I, E II, E III, E IV e E V). A ingestão de ração e o ganho de peso foram comparados ao final de 5 dias (ANOVA; Tukey-Kramer). No total, 7 grupos de 15 animais cada foram comparados. Para a faixa de 22°C foram utilizados 3 grupos, sendo um grupo experimental e dois grupos controle (CI e C II). Um deles foi mantido em condições ambientais semelhantes a biotérios convencionais sob ventilação geral diluidora (VGD) - C I. O outro grupo controle (C II) foi mantido no interior do equipamento de ventilação microambiental, porém, sem o direcionamento de ar, simulando a VGD. Os resultados obtidos demonstraram claramente que animais mantidos sob ventilação microambiental direta a 26°, 28° e 30° (E III, E IV e E V) apresentaram o mesmo ganho de massa corpórea que animais do grupo C I (22 &plusmn; 2°C). Os grupos E I e E II apresentaram menor ganho de massa corpórea quando comparados a C I (p<0,001 em ambas comparações). O ganho de peso de todos os grupos experimentais apresentou diferença estatística, quando comparado ao C II, exceto o grupo E V que obteve índice de ganho de peso equivalente a C II. A ingestão de ração de todos os grupos se manteve praticamente constante. O grupo E V apresentou uma redução na ingestão de ração quando comparado aos grupos C I, E I e E II (p<0,01; p<0,01; p<0,001 respectivamente). O grupo E III ingeriu menos ração que os grupos C I (p<0,05) e E II (p<0,05). / An air-conditioner for animal facility was assembled with Peltier effect thermoelectric modules. For external exchanger, had been tested: 1. external air dissipation: &delta;t = 14°C; 16,46% of efficiency; 1212 W/h of power consumption and, 2. external water dissipation: &delta;t = 21°C; 46,02% of efficiency; 524 W/h of power consumption. A mathematical simulation of operation, with not conditional air mixture, showed that the system can supply, with air dissipation, to &asymp; 91 microisolator rat cages and to &asymp; 137, with water dissipation. When compared with a freon system, the thermoelectric system shows economy of 26% in implantation and 38% in the electric consumption by generated BTU. The thermoelectric system showed too, a precision of ± 0,1°C, at experimental temperatures, what is impossible in a freon system. For animal tests, Wistar rats had been kept individually, in metabolic wire cages, without shelter, in microenvironmental system, under direct air flow at 0,6 m/s, under temperatures of 22°, 24°, 26°, 28° and 30° C (E I, E II, E III, E IV and E V). The food ingestion and the weight gain had been compared in the end of 5 days (ANOVA; Tukey-Kramer). In the total, 7 groups, 15 animals each, had been compared. For the 22°C temperature, had been used 3 groups, one experimental and two controls (C I e C II). One of them was kept in similar ambient of conventional laboratory animal rooms conditions (general diluitory ventilation, GDV) - C I. The other control group (C II) was kept in the interior of the equipment of microenvironmental ventilation, however, without the direct air flow, simulating the GDV. The gotten results demonstrate clearly that animal kept under direct microenvironmental ventilation at 26°, 28° and 30°C (E III, E IV and E V) have the same gain of corporal mass that C I group (22 &plusmn; 2°C). The E I and E II had less corporal mass gain when compared to C I (p<0,001 for the two comparisons). The weight gain for all the experimental groups, when compared to C II, presents statistical differences, except E V group, that was equal to C II. The food ingestion of all the groups was constant. The E V group presented a reduction in the food ingestion when compared with the groups C I , E I and E II (p<0,01; p<0,01; p<0,001 respectively). The E III group ingested less ration than C I (p<0,05) and E II (p<0,05) groups.

Apesar dos inúmeros grupos de pesquisa de todo o mundo investigando o câncer, esta doença tem se ampliado significativamente. No caso do câncer de ovário, a despeito dos notáveis avanços observados nos tratamentos atuais, um desafio persiste: a incapacidade da preservação de oócitos em tratamento com quimioterapia, radioterapia e/ou cirurgia, nos quais são freqüentes os efeitos colaterais tais como a perda folicular, a infertilidade, a menopausa precoce e as falências ovarianas pós-quimioterapia, decorrentes da irradiação e pós-cirurgia. Neste sentido, o transplante de tecido ovariano pode representar uma alternativa na recuperação da capacidade fértil da mulher, bem como uma terapia de reposição hormonal após tratamento. No presente estudo, foram utilizadas linhagens de camundongos para avaliar a viabilidade do transplante ovariano antes e após a irradiação, como alternativa para a restauração das funções reprodutivas. Para tanto, fêmeas das linhagens isogênicas C57BL/6/Unib e híbridas B6CF1/Unib, foram acasaladas com machos da linhagem C57BL/6/Unib, empregandose diferentes protocolos com fêmeas submetidas ou não aos efeitos da radiação gama de Co-60 nas doses de 4 Gy e 6 Gy. Os resultados demonstram que a transferência ovariana restabeleceu a capacidade reprodutiva dos animais para níveis muito próximos dos normais; que o tamanho médio da ninhada, não revelou diferenças significativas após a transferência ovariana; que os fragmentos do ovário da receptora permanecem funcionais em períodos próximos pós à irradiação com 4 Gy e 6 Gy e que a irradiação nas doses de 4 Gy e 6 Gy destroem as células germinativas comprometendo a reprodução. / Despite numerous research groups, around the world, investigating the cancer, the disease has expanded significantly. In the case of the ovarian cancer, in spite of the notable advances seen in current treatments, a challenge remains: the incapacity to cryopreserve the oocyte in cancer chemotherapy, radiotherapy or surgery, in which side effects are frequent such as follicular loss, infertility, early menopause and ovarian bankruptcy after chemotherapy, after irradiation and post-surgery. In this regard, the transplant of ovarian tissues may be an alternative in recovering the women fertility, as well a hormone replacement therapy after treatment. In our study, we used inbred strains and hybrid mice to assess the viability of ovarian transplantation before and after irradiation, as an alternative for the restoration of the reproductive functions. For that female from the C57BL/6/Unib inbred strain and from the hybrid, B6C-F1/Unib mated with C57BL/6/Unib males, employing different protocols with irradiated and non irradiated females with doses of 4 Gy and 6 Gy of Co60 gamma radiation. The results demonstrate that the ovarian transfer restored the reproductive capacity of the females to almost the normal levels; that the litter size average (production index) showed no significant differences after the ovarian transfer; that the fragments from the ovary of the recipient female are functional in periods coming after irradiation with 4 Gy and 6 Gy and that the irradiation with 4 Gy and 6 Gy destroys the germ cells, compromising the reproduction.

Thesis (PhD (Biomedical Sciences))--Cape Peninsula University of Technology, 2017. / The use of medicinal plants in the management of various health problems date back to the ancient times. However, only in recent years, researchers are starting to focus on the use of natural plant products as alternative treatment in disease control. Basella alba (Ba), commonly called Ceylon or Indian spinach is one of such medicinal plants, wildly cultivated and consumed mostly as vegetable. Studies have established many beneficial effects of Ba, including androgenic effects as well as antidiabetic effects which have been described in rats following oral administration of the leave extract. However, the actual mechanisms underlying the antihyperglyceamic effect of Ba have not been reported in any study and little or no research details are yet available on the potential beneficial effects of Ba in reproductive dysfunction resulting from diabetes mellitus. This study was aimed at investigating the mechanisms underlying the antidiabetic effect of Ba and the possibility of a role for the plant in correcting diabetic-induced reproductive dysfunctions in male Wistar rats. The first part of the study involved comparing of three different solvent extracts of Ba leaves namely ethyl acetate, methanolic and aqueous extracts for their antioxidant potentials, after which the aqueous extract was selected for further use in the experiments. Animal experimentation involved male rats (n=40) aged 8-10 weeks, randomly divided into four equal groups as follows: Healthy Control, Diabetic Control, Healthy Treatment and Diabetic Treatment. Diabetes was induced via a single intraperitoneal injection of streptozotocin (55mg/kg) and all animals subsequently received treatment via gavage (Rats in Control groups received 0.5ml/100g normal saline daily and treatment groups received 200mg/kg plant extract daily) for a period of four weeks. Fasting blood sugar and body weights were recorded weekly throughout the study. Animals were sacrificed upon completion of the treatment and blood samples and tissues collected for further analysis which included computer aided sperm analysis, Luminex® technology and enzyme-linked immunosorbent hormonal assays, inflammatory cytokine assays, analysis of oxidative stress markers and Histopathological analysis. The single intraperitoneal injection of a high streptozotocin dose resulted in hyperglycaemia, weight loss, subnormal sperm parameters, negative balance of inflammatory cytokines and endogenous antioxidants and degenerative changes in the pancreas, testes and epididymis as observed in the diabetic control rats. Oral administration with the aqueous extract of Ba for four weeks in diabetic treatment rats led to a significant reduction in blood sugar and improvement of sperm parameters by modulating the production of gonadal hormones, in vivo antioxidants and inflammatory cytokines. There was also significant recovery of normal islet histology and reduction in testicular and epididymal degeneration in the diabetic treatment rats when compared to their diabetic control counterparts. It was concluded from the findings of this study that the antidiabetic and profertility effects of Ba are largely dependent on the modulation of in vivo production of antioxidants, gonadal hormones and inflammatory cytokines, probably stimulated by one or more phytochemical component(s) that can be isolated in the aqueous extract of the plant

Cheuk, Yau Chuk. / Thesis submitted in: Dec 2008. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 132-144). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 論文摘要 --- p.iii / PUBLICATIONS --- p.v / ACKNOWLEDGEMENT --- p.vi / LIST OF ABBREBIVIATIONS --- p.vii / INDEX FOR FIGURES --- p.x / INDEX FOR TABLES --- p.xiv / TABLE OF CONTENTS --- p.xv / Chapter CHAPTER ONE - --- INTRODUCTION / Chapter 1.1 --- "Joint function, structure and biochemistry" / Chapter 1.1.1 --- Function of joint --- p.1 / Chapter 1.1.2 --- Types of cartilage --- p.1 / Chapter 1.1.3 --- Composition and structure of articular cartilage --- p.2 / Chapter 1.1.4 --- The subchondral bone --- p.3 / Chapter 1.1.5 --- Maturation of articular cartilage and subchondral bone --- p.3 / Chapter 1.2 --- Osteochondral defect / Chapter 1.2.1 --- Clinical problem --- p.6 / Chapter 1.2.2 --- Spontaneous repair --- p.7 / Chapter 1.2.3 --- Current treatment strategies --- p.7 / Chapter 1.2.4 --- Limitations of current treatment strategies --- p.8 / Chapter 1.2.5 --- Treatments under development --- p.11 / Chapter 1.2.6 --- Potential and limitations in cell therapies --- p.14 / Chapter 1.3 --- The 3-D scaffold-free cartilage / Chapter 1.3.1 --- Fabrication of scaffold-free cartilage --- p.16 / Chapter 1.3.2 --- Scaffold-free cartilage for chondral / osteochondral defect repair --- p.18 / Chapter 1.3.3 --- Scaffold-free bioengineered chondrocyte pellet from our group --- p.20 / Chapter 1.3.4 --- BCP as a possible treatment for OCD --- p.21 / Chapter 1.4 --- The objectives of the study --- p.22 / Chapter 1.5 --- The study plan / Chapter 1.5.1 --- Design of the study --- p.23 / Chapter 1.5.2 --- Choice of animal model --- p.23 / Chapter 1.5.3 --- Selection of evaluation time points --- p.24 / Chapter 1.5.4 --- Choice and modification of histological scoring system --- p.24 / Chapter CHAPTER TWO - --- METHODOLOGY / Chapter 2.1 --- Preparation of reagents and materials for tissue culture and histology --- p.26 / Chapter 2.2 --- Creation of osteochondral defect model --- p.28 / Chapter 2.3 --- Synthesis of scaffold-free cartilage using 3-D chondrocyte pellet culture / Chapter 2.3.1 --- Isolation of rabbit costal chondrocytes --- p.31 / Chapter 2.3.2 --- Three-dimensional chondrocyte pellet culture --- p.31 / Chapter 2.3.3 --- BrdU labeling for cell fate tracing --- p.32 / Chapter 2.4 --- Further characterization of the 3-D scaffold-free chondrocyte pellet / Chapter 2.4.1 --- Gross appearance --- p.35 / Chapter 2.4.2 --- Cell viability / Chapter 2.4.2.1 --- Alamar blue reduction assay --- p.35 / Chapter 2.4.3 --- Preparation of samples for histology --- p.36 / Chapter 2.4.4 --- General morphology and histomorphology / Chapter 2.4.4.1 --- H&E staining --- p.36 / Chapter 2.4.5 --- Cartilage properties / Chapter 2.4.5.1 --- Safranin O /Fast Green staining --- p.37 / Chapter 2.4.5.2 --- Immunohistochemistry of type II collagen --- p.37 / Chapter 2.4.5.3 --- Immunohistochemistry of type I collagen --- p.38 / Chapter 2.4.6 --- Angiogenic properties / Chapter 2.4.6.1 --- Immunohistochemistry of VEGF --- p.40 / Chapter 2.4.7 --- Osteogenic properties / Chapter 2.4.7.1 --- ALP staining --- p.40 / Chapter 2.5 --- Implantation of scaffold-free cartilage into osteochondral defect model / Chapter 2.5.1 --- Surgical procedures --- p.41 / Chapter 2.5.2 --- Experimental groups --- p.42 / Chapter 2.6 --- Assessment of osteochondral defect healing / Chapter 2.6.1 --- Macroscopic evaluation --- p.43 / Chapter 2.6.2 --- Preparation of samples for histology --- p.43 / Chapter 2.6.3 --- Histology for general morphology / Chapter 2.6.3.1 --- H&E staining --- p.45 / Chapter 2.6.4 --- Histological scoring / Chapter 2.6.4.1 --- Modification of the scoring system --- p.45 / Chapter 2.6.4.2 --- Procedures of scoring and validation --- p.45 / Chapter 2.6.5 --- Cell proliferation / Chapter 2.6.5.1 --- Immunohistochemistry of PCNA --- p.49 / Chapter 2.6.6 --- Cartilage regeneration / Chapter 2.6.6.1 --- Safranin O /Fast Green staining --- p.49 / Chapter 2.6.6.2 --- Immunohistochemistry of type II collagen --- p.49 / Chapter 2.6.6.3 --- Immunohistochemistry of type I collagen --- p.50 / Chapter 2.6.6.4 --- Polarized light microscopy --- p.50 / Chapter 2.6.7 --- Expression of angiogenic factor / Chapter 2.6.7.1 --- Immunohistochemistry of VEGF --- p.50 / Chapter 2.6.8 --- Bone regeneration / Chapter 2.6.8.1 --- μCT analysis --- p.50 / Chapter 2.6.9 --- Histomorphometric analysis of cartilage and bone regeneration --- p.53 / Chapter 2.6.10 --- BrdU detection for cell fate tracing --- p.55 / Chapter 2.6.11 --- Statistical analysis --- p.55 / Chapter CHAPTER THREE - --- RESULTS / Chapter 3.1 --- Further characterization of the 3-D chondrocyte pellet culture / Chapter 3.1.1 --- Gross examination --- p.57 / Chapter 3.1.2 --- Cell viability --- p.57 / Chapter 3.1.3 --- Cartilage properties --- p.61 / Chapter 3.1.4 --- Angiogenic properties --- p.63 / Chapter 3.1.5 --- Osteogenic properties --- p.64 / Chapter 3.2 --- Implantation of scaffold-free cartilage and assessment / Chapter 3.2.1 --- Gross examination --- p.65 / Chapter 3.2.2 --- General morphology --- p.67 / Chapter 3.2.3 --- Histological scores --- p.71 / Chapter 3.2.4 --- Cell proliferation --- p.75 / Chapter 3.2.5 --- Cartilage regeneration --- p.78 / Chapter 3.2.6 --- Expression of angiogenic factor --- p.90 / Chapter 3.2.7 --- Bone regeneration --- p.93 / Chapter 3.2.8 --- Histomorphometric analysis on cartilage and bone regeneration --- p.96 / Chapter 3.2.9 --- Cell fate tracing --- p.100 / Chapter CHAPTER FOUR - --- DISCUSSION / Chapter 4.1 --- Summary of key findings / Chapter 4.1.1 --- Further characterization of BCP and determination of implantation time --- p.102 / Chapter 4.1.2 --- Implantation of BCP in OCD --- p.102 / Chapter 4.2 --- Spontaneous healing in osteochondral defect / Chapter 4.2.1 --- Findings from the current study --- p.104 / Chapter 4.2.2 --- Comparison with other studies --- p.104 / Chapter 4.2.3 --- Factors affecting spontaneous healing --- p.105 / Chapter 4.3 --- Fabrication and further characterization of the 3-D chondrocyte pellet / Chapter 4.3.1 --- Comparison of different methods of producing scaffold-free cartilage construct --- p.106 / Chapter 4.3.2 --- Cartilage phenotype of the BCP --- p.107 / Chapter 4.3.3 --- Angiogenic and osteogenic potential of the BCP --- p.108 / Chapter 4.3.4 --- Role of mechanical stimulation on tissue-engineered cartilage --- p.109 / Chapter 4.4 --- Repair of osteochondral defect with allogeneic scaffold-free cartilage / Chapter 4.4.1 --- Advantages of the current scaffold-free chondrocyte pellet --- p.111 / Chapter 4.4.2 --- Remodeling of BCP after implantation --- p.111 / Chapter 4.4.3 --- Effect of BCP on cartilage repair --- p.112 / Chapter 4.4.4 --- Effect of BCP on bone regeneration / Chapter 4.4.4.1 --- Findings in the present study --- p.113 / Chapter 4.4.4.2 --- Possible reasons of slow bone repair --- p.114 / Chapter 4.4.4.3 --- Effect of BCP on bone region peripheral to defect --- p.115 / Chapter 4.4.5 --- Immunorejection-free properties of the BCP --- p.116 / Chapter 4.4.6 --- Comparison with other animal studies using scaffold-free cartilage --- p.117 / Chapter 4.4.7 --- Possibility of implanting a BCP cultured for shorter or longer period --- p.118 / Chapter 4.4.8 --- Scaffold-free cartilage construct and construct with scaffold for OCD repair --- p.119 / Chapter 4.4.9 --- Chondrocytes and stem cells for OCD repair --- p.120 / Chapter 4.5 --- Limitations of the study / Chapter 4.5.1 --- Animal model --- p.122 / Chapter 4.5.2 --- Histomorphometric analysis --- p.122 / Chapter 4.5.3 --- Lack of quantitative data analysis --- p.122 / Chapter 4.5.4 --- BrdU labeling of cells --- p.123 / Chapter 4.5.5 --- Lack of biomechanical test --- p.123 / Chapter 4.5.6 --- Small sample size --- p.123 / Chapter CHAPTER FIVE - --- CONCLUSION --- p.124 / Chapter CHAPTER SIX - --- FUTURE STUDIES / Chapter 6.1 --- Identification of factors affecting bone repair after OCD treatment --- p.125 / Chapter 6.2 --- Modifications of BCP treatment --- p.125 / Chapter 6.3 --- Alternative cell source --- p.126 / Chapter 6.4 --- Alternative cell tracking methods --- p.126 / Chapter 6.5 --- Inclusion of biomechanical test --- p.126 / APPENDICES / Appendix 1. Conference paper 1 --- p.129 / Appendix 2: Conference paper 2 --- p.130 / Appendix 3: Animal experimentation ethics approval --- p.131 / BIBLIOGRAPHY --- p.132