Yes / Metabolic syndrome is a global public health problem and predisposes individuals to obesity, diabetes and cardiovascular disease. Although the underlying mechanisms remain to be elucidated, accumulating evidence has uncovered a critical role of adipokines. Chemerin, encoded by the gene Rarres2, is a newly discovered adipokine involved in inflammation, adipogenesis, angiogenesis and energy metabolism. In humans, local and circulating levels of chemerin are positively correlated with body mass index and obesity-related biomarkers. In this review, we discuss both peripheral and central roles of chemerin in regulating body metabolism. In general, chemerin is upregulated in obese and diabetic animals. Previous studies by gain or loss of function show an association of chemerin with adipogenesis, glucose homeostasis, food intake and body weight. In the brain, the hypothalamus integrates peripheral afferent signals including adipokines to regulate appetite and energy homeostasis. Chemerin increases food intake in seasonal animals by acting on hypothalamic stem cells, the tanycytes. In peripheral tissues, chemerin increases cell expansion, inflammation and angiogenesis in adipose tissue, collectively resulting in adiposity. While chemerin signalling enhances insulin secretion from pancreatic islets, contradictory results have been reported on how chemerin links to obesity and insulin resistance. Given the association of chemerin with obesity comorbidities in humans, advances in translational research targeting chemerin are expected to mitigate metabolic disorders. Together, the exciting findings gathered in the last decade clearly indicate a crucial multifaceted role for chemerin in the regulation of energy balance, making it a promising candidate for urgently needed pharmacological treatment strategies for obesity.

Yes / Chemerin is an adipokine involved in inflammation, adipogenesis, angiogenesis and energy metabolism, and has been hypothesized as a link between obesity and type II diabetes. In humans affected by obesity, chemerin gene expression in peripheral tissues and circulating levels are elevated. In mice, plasma levels of chemerin are upregulated by high-fat feeding and gain and loss of function studies show an association of chemerin with body weight, food intake and glucose homeostasis. Therefore, chemerin is an important blood-borne mediator that, amongst its other functions, controls appetite and body weight. Almost all studies of chemerin to date have focused on its release from adipose tissue and its effects on peripheral tissues with the central effects largely overlooked. To demonstrate a central role of chemerin, we manipulated chemerin signaling in the hypothalamus, a brain region associated with appetite regulation, using pharmacological and genetic manipulation approaches. Firstly, the selective chemerin receptor CMKLR1 antagonist α-NETA was administered i.c.v. to rats to test for an acute physiological effect. Secondly, we designed a short-hairpin-RNA (shRNA) lentivirus construct targeting expression of CMKLR1. This shRNA construct, or a control construct was injected bilaterally into the arcuate nucleus of male Sprague Dawley rats on high-fat diet (45%). After surgery, rats were maintained on high-fat diet for 2 weeks and then switched to chow diet for a further 2 weeks. We found a significant weight loss acutely and inhibition of weight gain chronically. This difference became apparent after diet switch in arcuate nucleus-CMKLR1 knockdown rats. This was not accompanied by a difference in blood glucose levels. Interestingly, appetite-regulating neuropeptides remained unaltered, however, we found a significant reduction of the inflammatory marker TNF-α suggesting reduced expression of CMKLR1 protects from high-fat diet induced neuroinflammation. In white and brown adipose tissue, mRNA expression of chemerin, its receptors and markers of adipogenesis, lipogenesis and brown adipocyte activation remained unchanged confirming that the effects are driven by the brain. Our behavioral analyses suggest that knockdown of CMKLR1 had an impact on object recognition. Our data demonstrate that CMKLR1 is functionally important for the central effects of chemerin on body weight regulation and neuroinflammation. / This work was funded in part by the Academy of Medical Sciences, the Wellcome Trust, the Government of Business, Energy and Industrial Strategy and the British Heart Foundation and Diabetes United Kingdom [SBF004/1063] (GH), the Society for Endocrinology Equipment Grant (GH, RD), the University of Bradford (GH, KP, SK) and Nottingham Trent University (RD).

Iveta Šteflová Drug induced imbalance of sodium Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Department of Biological and Medical Science Supervisor: Doc.MUDr. Josef Herink, DrSc. Sodium (Na+ ) is the major extracellular cation. It plays an important role in maintaining membrane potential and depolarization that is the basic mechanism of transmission of the nerve impulse. It is involved in maintaining acid-base balance, osmotic pressure, water retention in the body. The largest part of the sodium is in the extracellular fluid where it is stored about 50 % of sodium. Plasma concentration of sodium is 140 ± 5 mmol/l. Drug-induced electrolyte disorders are increasingly reported and may be associated with considerable morbidity and mortality. The risk of drug-induced hyponatremia is generally higher than the risk of drug-induced hypernatremia. Hyponatremia is a common electrolyte disorder defined as a decrease plasma sodium concentration below 135 mmol/l. It is classified by the state of volume - hypovolemic, euvolemic and hypervolemic hyponatremia. It reflects the relative rate between sodium and water in the body. The most common cause is the syndrome of inappropriate secretion of antidiuretic hormone that induces euvolemic hyponatremia. Hypernatremia is...

Hepcidin is a well-known iron-regulatory hormone that plays a key role in maintaining peripheral iron homeostasis. The presence and wide-spread distribution of hepcidin in the brain suggests that this peptide may also be an important player in brain iron homeostasis. In this study, we tested the hypothesis that hepcidin exerts an important role in the regulation of brain iron content, which might benefit iron-associated NDs such as PD. We also examined the hypothesis that hepcidin could control iron transport processes via regulating iron transport proteins in the brain cells, thus maintaining brain iron homeostasis. / In conclusion, the results of the present study implied that hepcidin plays an important role in maintaining brain iron homeostasis. Hepcidin is beneficial for PD and this effect is related to its iron-regulatory effect via inhibiting iron accumulation in the substantia nigra. Hepcidin effectively controls iron uptake and release through regulating iron transport proteins expressions in the brain, which would contribute to brain iron homeostasis. Therefore, manipulation of hepcidin level in the brain has a potential to be developed into a novel preventive approach for the iron-associated NDs such as PD. / In the second part, we investigated the effect of hepcidin on the processes of iron uptake and release in the cultured brain cells including neurons, astrocytes and brain vascular endothelial cells (BVECs). The expressions of iron uptake proteins such as transferrin receptor 1 (TfR1) and divalent metal transporter 1 (DMT1) as well as the iron exporter ferroportin 1 (Fpn1) were also observed. We found that hepcidin reduced both iron uptake and release via decreasing iron transport proteins expressions in these brain cells, which would contribute to its iron regulatory effect. Finally, we further explored the mechanisms underlying the regulatory effect of hepcidin on the iron transporters in the last part, and found that the action of hepcidin in reducing TfR1 expression is a direct and cAMP-PKA (Cyclic Adenosine 3', 5'-monophosphate/ Protein Kinase-A) pathway-dependent event. / Iron is a transition trace metal essential for mammalian cellular and tissue viability. It also plays important roles in the central nervous system (CNS), including embryonic brain development, myelination, and neurotransmitters synthesis. However, abnormal iron accumulation has been demonstrated in a number of neurodegenerative diseases (NDs) such as Parkinson's (PD), Alzheimer's (AD) and Huntington's diseases (HD). Currently very little is known about the mechanisms involved in brain iron homeostasis and therefore it is not known why and how iron is abnormally increased in the brain. However, given the essential role that excess iron plays in the pathological processes in the NDs, to suppress the accumulated iron is expected to be an effective strategy to prevent and treat these NDs. / To investigate whether hepcidin could benefit iron-associated NDs including PD and whether this beneficial role is related to its iron-regulatory function in the brain, in the first part of study, we investigated the effects of hepcidin on the 6-hydroxydopamine (6-OHDA) in vitro and in vivo PD models. We found that in primary cultured mesencephalic (MES) neurons, hepcidin overexpression via adenovirus-hepcidin (Ad-hepcidin) infection prevented 6-OHDA-induced increase in cellular iron content and protected the MES neurons. In the 6-OHDA model of PD in vivo, overexpression of hepcidin in the substantia nigra via Ad-hepcidin intranigral injection significantly prevented iron accumulation and dopaminergic neurons loss in the pars compacta of substantia nigra (SNc). These effects were accompanied by a marked improvement in motor performance of the PD animals. These findings indicate that hepcidin could benefit iron-associated NDs such as PD and via its iron-regulatory role in the brain. / Du, Fang. / Adviser: Ya Ke. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 152-173). / 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.

Li, Sui Mui. / On t.p. "beta" appears as the Greek letter. / Thesis submitted in: December 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 182-189). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese) --- p.iii / Acknowledgements --- p.v / Table of contents --- p.vi / List of figures --- p.xii / List of appendices --- p.xix / Abbreviations --- p.xx / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter Chapter 2 --- Role of PPARP in adipocyte differentiation - an in vitro study --- p.20 / Chapter 2.1 --- Introduction --- p.21 / Chapter 2.2 --- Materials and Methods --- p.23 / Chapter 2.2.1 --- Preparation ofPPARβ (+/+) and PPARβ (-/-) MEFs --- p.23 / Chapter 2.2.1.1 --- Materials --- p.23 / Chapter 2.2.1.2 --- Methods --- p.23 / Chapter 2.2.1.2.1 --- Isolation of MEFs --- p.23 / Chapter 2.2.1.2.2 --- Passage ofMEF culture --- p.25 / Chapter 2.2.2 --- Genotyping of PPARβ (+/+) and PPARβ (-/-) MEFs --- p.25 / Chapter 2.2.2.1 --- Materials --- p.26 / Chapter 2.2.2.2 --- Methods --- p.26 / Chapter 2.2.2.2.1 --- Primer design --- p.26 / Chapter 2.2.2.2.2 --- Genomic DNA extraction --- p.27 / Chapter 2.2.2.2.3 --- PCR reaction --- p.29 / Chapter 2.2.3 --- Western blotting of PPARβ(+/+) and PPARβ (-/-) MEFs --- p.30 / Chapter 2.2.3.1 --- Materials --- p.30 / Chapter 2.2.3.2 --- Methods --- p.31 / Chapter 2.2.3.2.1 --- Preparation of nuclear extracts --- p.31 / Chapter 2.2.3.2.2 --- Western blot --- p.32 / Chapter 2.2.4 --- Induction of adipocyte differentiation of PPARβ (+/+) and PPARβ(-/-) MEFs --- p.33 / Chapter 2.2.4.1 --- Materials --- p.34 / Chapter 2.2.4.2 --- Methods --- p.34 / Chapter 2.2.4.2.1 --- Seeding ofMEFs --- p.34 / Chapter 2.2.4.2.2 --- Adipocyte differentiation --- p.35 / Chapter 2.2.5 --- Oil Red O staining of differentiated PPARβ(+/+) and PPARβ(-/-) MEFs --- p.36 / Chapter 2.2.5.1 --- Materials --- p.36 / Chapter 2.2.5.2 --- Method --- p.37 / Chapter 2.2.5.2.1 --- Oil Red O staining --- p.37 / Chapter 2.2.6 --- Determination of triglyceride-protein assay of differentiated PPARβ (+/+) and PPARβ (-/-) MEFs --- p.37 / Chapter 2.2.6.1 --- Materials --- p.39 / Chapter 2.2.6.2 --- Methods --- p.39 / Chapter 2.2.6.2.1 --- Lysis of differentiated MEFs --- p.39 / Chapter 2.2.6.2.2 --- Measurement of triglyceride concentration in cell lysate --- p.40 / Chapter 2.2.6.2.3 --- Measurement of protein concentration in cell lysate --- p.41 / Chapter 2.2.7 --- Preparation of PPARβ(+/+) and PPARβ (-/-) MEF RNA for RT-PCR and Northern blot analysis --- p.42 / Chapter 2.2.7.1 --- Materials --- p.42 / Chapter 2.2.7.2 --- Method --- p.42 / Chapter 2.2.7.2.1 --- RNA isolation --- p.42 / Chapter 2.2.8 --- RT-PCR analysis of differentiated PPARβ(+/+) and PPARβ (-/-) MEFs --- p.44 / Chapter 2.2.8.1 --- Materials --- p.45 / Chapter 2.2.8.2 --- Methods --- p.45 / Chapter 2.2.8.2.1 --- Primer design --- p.45 / Chapter 2.2.8.2.2 --- RT-PCR --- p.46 / Chapter 2.2.9 --- Northern blot analysis of differentiated PPARβ(+/+) and PPARβ (-/-) MEFs --- p.47 / Chapter 2.2.9.1 --- Materials --- p.48 / Chapter 2.2.9.2 --- Methods --- p.49 / Chapter 2.2.9.2.1 --- Preparation of cDNA probes for Northern blotting --- p.49 / Chapter 2.2.9.2.1.1 --- RNA extraction --- p.49 / Chapter 2.2.9.2.1.2 --- Primer design --- p.49 / Chapter 2.2.9.2.1.3 --- RT-PCR of extracted mRNA --- p.50 / Chapter 2.2.9.2.1.4 --- Subcloning of amplified cDNA products --- p.50 / Chapter 2.2.9.2.1.5 --- Screening of recombinant clones by phenol-chloroform extraction --- p.51 / Chapter 2.2.9.2.1.6 --- Confirmation of the recombinant clones by restriction enzyme site mapping --- p.52 / Chapter 2.2.9.2.1.7 --- Confirmation of the recombinant clones by PCR method --- p.52 / Chapter 2.2.9.2.1.8 --- Mini-preparation of plasmid DNA from the selected recombinant clones --- p.54 / Chapter 2.2.9.2.1.9 --- Preparation of cDNA probes --- p.54 / Chapter 2.2.9.2.1.10 --- Formaldehyde agarose gel electrophoresis of RNA --- p.55 / Chapter 2.2.9.2.1.11 --- Hybridization and color development --- p.56 / Chapter 2.3 --- Results --- p.58 / Chapter 2.3.1 --- Confirmation of PPARβ(+/+) and PPARβ (-/-) MEFs genotypes --- p.58 / Chapter 2.3.2 --- PPARβ (-/-) MEFs differentiated similarly to PPARβ(+/+) MEFs as measured by Oil Red O staining --- p.61 / Chapter 2.3.3 --- PPARβ (-/-) MEFs differentiated similarly to PPARβ(+/+) MEFs as reflected by their intracellular triglyceride contents --- p.64 / Chapter 2.3.4 --- PPARβ(-/-) MEFs expressed the adipocyte differentiation marker genes similarly to PPARβ (+/+) MEFs --- p.66 / Chapter 2.4 --- Discussion --- p.77 / Chapter Chapter 3 --- Role of PPARβ in adipocyte differentiation and lipid homeostasis - an in vivo study --- p.82 / Chapter 3.1 --- Introduction --- p.83 / Chapter 3.2 --- Materials and Methods --- p.85 / Chapter 3.2.1 --- Animal and high fat diet treatment --- p.85 / Chapter 3.2.1.1 --- Materials --- p.85 / Chapter 3.2.1.2 --- Method --- p.86 / Chapter 3.2.1.2.1 --- Animal treatment --- p.86 / Chapter 3.2.2 --- Tail-genotyping of PPARβ (+/+) and PPARβ (-/-) mice --- p.87 / Chapter 3.2.2.1 --- Materials --- p.87 / Chapter 3.2.2.2 --- Methods --- p.88 / Chapter 3.2.2.2.1 --- DNA extraction from tail --- p.88 / Chapter 3.2.2.2.2 --- PCR tail-genotyping --- p.89 / Chapter 3.2.3 --- "Measurement of serum triglyceride, cholesterol and glucose levels by enzymatic and spectrophometric methods" --- p.89 / Chapter 3.2.3.1 --- Materials --- p.90 / Chapter 3.2.3.2 --- Methods --- p.91 / Chapter 3.2.3.2.1 --- Serum preparation --- p.91 / Chapter 3.2.3.2.2 --- Measurement of serum triglycerides --- p.91 / Chapter 3.2.3.2.3 --- Measurement of serum cholesterol --- p.92 / Chapter 3.2.3.2.3 --- Measurement of serum glucose --- p.93 / Chapter 3.2.4 --- Measurement of serum insulin and leptin levels by ELISA --- p.94 / Chapter 3.2.4.1 --- Materials --- p.95 / Chapter 3.2.4.2 --- Methods --- p.95 / Chapter 3.2.4.2.1 --- Measurement of serum insulin --- p.95 / Chapter 3.2.4.2.2 --- Measurement of serum leptin --- p.97 / Chapter 3.2.5 --- "Histological studies of liver, interscapular BF and gonadal WF pads" --- p.99 / Chapter 3.2.5.1 --- Materials --- p.100 / Chapter 3.2.5.2 --- Methods --- p.100 / Chapter 3.2.5.2.1 --- "Fixation, dehydration, embedding in paraffin and sectioning" --- p.100 / Chapter 3.2.5.2.2 --- H&E staining --- p.101 / Chapter 3.2.6 --- Analyses of fecal lipid contents --- p.102 / Chapter 3.2.6.1 --- Materials --- p.102 / Chapter 3.2.6.2 --- Method --- p.103 / Chapter 3.2.6.2.1 --- Extraction of lipid contents from stools --- p.103 / Chapter 3.2.7 --- Statistical analysis --- p.104 / Chapter 3.3 --- Results --- p.105 / Chapter 3.3.1 --- Confirmation of genotypes by PCR --- p.105 / Chapter 3.3.2 --- PPARβ (-/-) mice were more resistant to high fat diet-induced obesity --- p.105 / Chapter 3.3.3 --- PPARβ (-/-) mice consumed similarly as to PPARβ (+/+) counterparts… --- p.122 / Chapter 3.3.4 --- Effect of high fat diet on organ weights --- p.128 / Chapter 3.3.4.1 --- PPARβ (-/-) mice were more resistant to high fat diet-induced liver hepatomegaly --- p.134 / Chapter 3.3.4.2 --- PPARβ (-/-) mice were resistant to high fat diet-induced increased white fat depots --- p.134 / Chapter 3.3.4.3 --- PPARβ (-/-) mice were resistant to high fat diet-induced increased brown fat mass --- p.137 / Chapter 3.3.5 --- Effect of high fat diet on organ histology --- p.142 / Chapter 3.3.5.1 --- PPARβ(-/-) mice were more resistant to high fat diet-induced liver steatosis --- p.143 / Chapter 3.3.5.2 --- No defect in white adipocyte expansion in PPARβ(-/-) mice upon high fat diet feeding --- p.153 / Chapter 3.3.5.3 --- No defect in brown adipocyte expansion in PPARβ (-/-) mice upon high fat diet feeding --- p.159 / Chapter 3.3.6 --- "Effect on high fat diet on serum cholesterol, triglyceride, glucose, insulin and leptin levels" --- p.164 / Chapter 3.3.6.1 --- "PPARβ (-/-) mice had a lower serum cholesterol level, but a similar triglyceride level as compared to PPARβ (+/+) mice upon high fat diet feeding" --- p.165 / Chapter 3.3.6.2 --- PPARβ (-/-) mice were resistant to high fat diet-induced insulin resistance --- p.167 / Chapter 3.3.6.3 --- PPARβ (-/-) mice had a similar serum leptin level as PPARβ (+/+) mice --- p.170 / Chapter 3.3.7 --- No decision made in fecal lipid content of PPARβ (+/+) and PPARβ (-/-) mice --- p.173 / Chapter 3.4 --- Discussion --- p.176 / References --- p.182 / Appendices --- p.190

This dissertation is an initial study of how modern engineering control may be applied to reverse engineer homeostasis in metabolic pathways using high-throughput biological data. This attempt to reconcile differences between engineering control and biological homeostasis from an interdisciplinary perspective is motivated not only by the observation that robust behavior in metabolic pathways resembles stabilized dynamics in controlled systems, but also by the challenges forewarned in achieving a true meeting of minds between engineers and biologists. To do this, a comparator model is developed and applied to model the effect of single-gene (SPT) overexpression on C16:0 sphingolipid de novo biosynthesis in vitro, specifically to simulate and predict potential homeostatic pathway interactions between the sphingolipid metabolites. Sphingolipid de novo biosynthesis is highly regulated because its pathway intermediates are highly bioactive. Alterations in sphingolipid synthesis, storage, and metabolism are implicated in human diseases. In addition, when variation in structure is considered, sphingolipids are one of the most diverse and complex families of biomolecules. To complete the modeling paradigm, wild type cells are defi ned as the reference that exhibits the "desired" pathway dynamics that the treated cells approach. Key model results show that the proposed modern engineering control approach using a comparator to reverse engineer homeostasis in metabolic systems is: (a) eff ective in capturing observed pathway dynamics from experimental data, with no signifi cant di fference in precision from existing models, (b) robust to potential errors in estimating state-space parameters as a result of sparse data, (c) generalizable to model other metabolic systems, as demonstrated by testing on a separate independent dataset, and (d) biologically relevant in terms of predicting steady-state feedback as a result of homeostasis that is verifi ed in literature and with additional independent data from drug dosage experiments.

The mammalian brain is the most cholesterol-rich organ of the body, requiring in situ de novo cholesterol synthesis to maintain its cholesterol requirement. Defects in brain cholesterol homeostasis are implicated in cognitive deficits related to aging and in neurodegenerative diseases such as Alzheimer's Disease (AD). Oxysterol-binding protein (OSBP) - related proteins are highly conserved cytosolic proteins that coordinate lipid homeostasis by regulating cell signaling, inter-organelle membrane contact sites and non-vesicular transport of cholesterol. Previously, ORP6, a poorly characterized member of this family, was found to be part of complex transcriptional cascade coordinated by SBREP2 and emerged as a novel regulator of intracellular cholesterol trafficking in hepatocytes and macrophages. Yet how ORP6 regulates these pathways and its function in the brain where it is most highly expressed is unknown. Here, we show that ORP6 is highly expressed in the brain, where it exhibits spatial and cell-type specific expression. ORP6 expression is enriched in the hippocampus and caudal-putamen brain regions, specifically within neurons and astrocytes. ORP6 knockdown in astrocytes altered the expression of cholesterol biosynthesis, cholesterol efflux and cholesterol esterification genes, resulting in the accumulation of esterified cholesterol within cytoplasmic lipid droplets and reduced cholesterol efflux highlighting a role for ORP6 in astrocytic cholesterol metabolism. We also present in this thesis, the newly generated second viable ORP family member knockout mouse. ORP6 ablation in mice results in the dysregulation of brain and whole-body lipid homeostasis, increased Aβ deposition in the brain and neuroanatomical alterations. Together, our findings highlight a critical role for cholesterol trafficking proteins in brain cholesterol homeostasis and identify ORP6 as a potential novel target for AD.

D.Diac. (Play Therapy) / Teachers in South Africa experience strain and tension, unique to their specific work description, which is evident in the career-related stress symptoms experienced by the individual teachers, the high rate of absenteeism amongst teachers, as well as the high attrition rate. Although factors causing teacher-stress, and the consequences thereof, have been extensively researched, a limited number of empirical evaluations of the effectiveness and accessibility of stress-management programmes have been conducted. Due to the holistic nature of Gestalt therapy, its emphasis on the here-and-now and the Gestalt principles of awareness, dialogue and process, a Gestalt therapeutic process model was developed to empower teachers to regain homeostasis. The aim and objectives of the research were the design, development, presentation and evaluation of a Gestalt therapeutic process model for teachers suffering from career-related stress, in their quest to regain homeostasis. The model was developed to be implemented within the school environment by a trained member of the school management team. The process of intervention research was used for the research study. A functional Gestalt therapeutic programme, based on the theoretical Gestalt therapeutic model, was developed and presented to ten teachers, selected through purposive sampling, and again to five different teachers, selected through theoretical sampling. The teachers identified were representative regarding age, gender, race and years in education. Triangulation was used and qualitative and quantitative data were collected simultaneously. The hypothesis stated for the research was that if teachers, suffering from career-related stress, were exposed to a Gestalt therapeutic model, they would regain homeostasis. Both the qualitative and quantitative data supported the hypothesis. The effect of the variables on each other was compared to confirm the reliability, applicability and neutrality of the research data. At the end of the three month research period the teachers who were exposed to the said model experienced less stress-related symptoms, as well as growth towards maturity and self-support, which would ultimately result in the regaining of homeostasis. A further objective of the research was to determine the feasibility of a trained school management team member implementing the Gestalt therapeutic process model at school. The qualitative data collected, indicated the feasibility thereof on condition that the school management team member did receive the necessary Gestalt therapeutic training. / Social Work

Wong Yin-Mei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 119-135). / Abstracts in English and Chinese. / Abstract (English version) --- p.i / Abstract (Chinese version) --- p.iii / Publications --- p.v / Acknowledgements --- p.vi / Table of contents --- p.viii / List of tables --- p.xi / List of figures --- p.xii / Abbreviations --- p.xiv / Chapter Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Osteoporosis --- p.1 / Chapter 1.1.1 --- Consensus statement --- p.1 / Chapter 1.1.2 --- Epidemiology and outcomes --- p.4 / Chapter 1.1.2.1 --- Hip fractures --- p.4 / Chapter 1.1.2.2 --- Vertebral fractures --- p.5 / Chapter 1.1.2.3 --- Wrist fractures --- p.7 / Chapter 1.1.3 --- Postmenopausal osteoporosis --- p.8 / Chapter 1.1.3.1 --- Pathogenesis --- p.8 / Chapter 1.1.3.1.1 --- Genetics --- p.11 / Chapter 1.1.3.1.2 --- Bone remodeling --- p.14 / Chapter 1.1.3.1.3 --- Calcium homeostasis --- p.21 / Chapter 1.1.3.1.4 --- Life style 一 nutrition and exercise --- p.26 / Chapter 1.1.3.2 --- Current pharmacological treatment --- p.27 / Chapter 1.1.3.2.1 --- Introduction --- p.27 / Chapter 1.1.3.2.2 --- Limitations --- p.31 / Chapter 1.2 --- Traditional Chinese medicine --- p.33 / Chapter 1.2.1 --- The Kidney --- p.33 / Chapter 1.2.2 --- Kidney-tonifying herbs --- p.33 / Chapter 1.3 --- Aim of the studies --- p.36 / Chapter Chapter 2. --- Materials and methods --- p.38 / Chapter 2.1 --- Kidney-tonifying herbs and herbal formula --- p.38 / Chapter 2.1.1 --- Sources --- p.38 / Chapter 2.1.2 --- Herbal extract preparation --- p.38 / Chapter 2.2 --- Animal study --- p.40 / Chapter 2.2.1 --- Reagents --- p.40 / Chapter 2.2.2 --- Animal care --- p.40 / Chapter 2.2.3 --- Herbs and herbal formula preparations for animal studies --- p.41 / Chapter 2.2.4 --- Experimental design --- p.41 / Chapter 2.2.5 --- Gene expression study --- p.44 / Chapter 2.2.5.1 --- Tissue preparation --- p.44 / Chapter 2.2.5.2 --- Isolation of total RNA --- p.45 / Chapter 2.2.5.3 --- Complementary DNA synthesis --- p.47 / Chapter 2.2.5.4 --- Real-time polymerase chain reaction analysis --- p.47 / Chapter 2.3 --- Cell culture study --- p.49 / Chapter 2.3.1 --- Reagents --- p.49 / Chapter 2.3.2 --- Cell lines --- p.49 / Chapter 2.3.2.1 --- "Rat osteosarcoma cell line, UMR-106" --- p.49 / Chapter 2.3.2.2 --- "Human breast cancer cell line, MCF-7" --- p.50 / Chapter 2.3.2.3 --- Cell culture techniques --- p.50 / Chapter 2.3.3 --- Herbs preparations for cell culture --- p.51 / Chapter 2.3.4 --- Cell viability assay --- p.51 / Chapter 2.3.5 --- Cellular alkaline phosphatase activity assay --- p.52 / Chapter 2.3.6 --- Matrix mineralization assay --- p.54 / Chapter 2.3.7 --- Competitive estrogen receptor binding assay --- p.56 / Chapter 2.4 --- Statistical analyses --- p.58 / Chapter Chapter 3. --- Results --- p.59 / Chapter 3.1 --- Extraction yields of Kidney-tonifying herbs and herbal formula --- p.59 / Chapter 3.2 --- Effects of Kidney-tonifying herbs and herbal formula on the gene expressions of calcium absorption and reabsorption related genes --- p.61 / Chapter 3.2.1 --- Gene expression of 25-hydroxyvitamin D3-1 alpha-hydroxylasein the kidney --- p.62 / Chapter 3.2.2 --- Gene expression of vitamin D receptor in the duodenum --- p.65 / Chapter 3.2.3 --- Gene expression of calbindin D9K in the duodenum --- p.67 / Chapter 3.2.4 --- Gene expression of vitamin D receptor in the kidney --- p.69 / Chapter 3.2.5 --- Gene expression of calbindin D28K in the kidney --- p.71 / Chapter 3.3 --- Effects of Kidney-tonifying herbs on osteoblastic UMR-106 cell line --- p.73 / Chapter 3.3.1 --- Effects of Kidney-tonifying herbs on the cell viability of UMR-106 cells --- p.73 / Chapter 3.3.2 --- Effects of Kidney-tonifying herbs on the osteoblastic differentiation of UMR-106 cells --- p.76 / Chapter 3.3.2.1 --- Cellular alkaline phosphatase activity --- p.76 / Chapter 3.3.2.2 --- Degree of matrix mineralization --- p.80 / Chapter 3.4 --- Estrogen receptor binding activities of Kidney-tonifying herbs --- p.85 / Chapter Chapter 4. --- Discussion --- p.89 / Chapter 4.1 --- Safety of Kidney-tonifying herbs and herbal formula --- p.89 / Chapter 4.2 --- Kidney-tonifying herbs and herbal formula preserve bone mineral density --- p.93 / Chapter 4.3 --- Kidney-tonifying herbs and herbal formula modulate calcium homeostasis --- p.97 / Chapter 4.3.1 --- "Roles in renal synthesis of the hormonally active form of vitamin D: 1,25-dihydroxyvitamin D3" --- p.97 / Chapter 4.3.2 --- Roles in calcium absorption in the duodenum --- p.99 / Chapter 4.3.3 --- Roles in calcium reabsorption in the kidney --- p.102 / Chapter 4.3.4 --- Summary --- p.104 / Chapter 4.4 --- Kidney-tonifying herbs modulate bone formation --- p.106 / Chapter 4.4.1 --- Effects on osteoblast proliferation --- p.106 / Chapter 4.4.2 --- Effects on osteoblastic differentiation --- p.107 / Chapter 4.4.3 --- Summary --- p.108 / Chapter 4.5 --- Kidney-tonifying herbs interact with estrogen receptor --- p.110 / Chapter 4.6 --- Active ingredients of Kidney-tonifying herbs --- p.111 / Chapter 4.7 --- Limitations of the present studies --- p.115 / Chapter 4.8 --- Conclusion and future prospect --- p.117 / References --- p.119

D.Diac. (Play Therapy) / Teachers in South Africa experience strain and tension, unique to their specific work description, which is evident in the career-related stress symptoms experienced by the individual teachers, the high rate of absenteeism amongst teachers, as well as the high attrition rate. Although factors causing teacher-stress, and the consequences thereof, have been extensively researched, a limited number of empirical evaluations of the effectiveness and accessibility of stress-management programmes have been conducted. Due to the holistic nature of Gestalt therapy, its emphasis on the here-and-now and the Gestalt principles of awareness, dialogue and process, a Gestalt therapeutic process model was developed to empower teachers to regain homeostasis. The aim and objectives of the research were the design, development, presentation and evaluation of a Gestalt therapeutic process model for teachers suffering from career-related stress, in their quest to regain homeostasis. The model was developed to be implemented within the school environment by a trained member of the school management team. The process of intervention research was used for the research study. A functional Gestalt therapeutic programme, based on the theoretical Gestalt therapeutic model, was developed and presented to ten teachers, selected through purposive sampling, and again to five different teachers, selected through theoretical sampling. The teachers identified were representative regarding age, gender, race and years in education. Triangulation was used and qualitative and quantitative data were collected simultaneously. The hypothesis stated for the research was that if teachers, suffering from career-related stress, were exposed to a Gestalt therapeutic model, they would regain homeostasis. Both the qualitative and quantitative data supported the hypothesis. The effect of the variables on each other was compared to confirm the reliability, applicability and neutrality of the research data. At the end of the three month research period the teachers who were exposed to the said model experienced less stress-related symptoms, as well as growth towards maturity and self-support, which would ultimately result in the regaining of homeostasis. A further objective of the research was to determine the feasibility of a trained school management team member implementing the Gestalt therapeutic process model at school. The qualitative data collected, indicated the feasibility thereof on condition that the school management team member did receive the necessary Gestalt therapeutic training. / Social Work