Man, Ka Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 101-126). / Abstracts in English and Chinese. / Chapter I. --- Title page --- p.i / Chapter II. --- Thesis committee --- p.ii / Chapter III. --- Acknowledgements --- p.iii / Chapter IV. --- Abstract (English) --- p.iv / Chapter V. --- Abstract (Chinese) --- p.vi / Chapter VI. --- Table of contents --- p.vii / Chapter VII. --- List of abbreviations --- p.xiii / Chapter VIII. --- List of figures --- p.xv / General introduction --- p.1 / Chapter Chapter 1: --- Literature review --- p.4 / Chapter 1.1. --- Cadmium --- p.5 / Chapter 1.1.1. --- Cadmium - Ways of uptake in human and aquatic life --- p.5 / Chapter 1.1.2. --- Cadmium - Toxic effects in fish --- p.6 / Chapter 1.2. --- Cortisol --- p.11 / Chapter 1.2.1. --- Cortisol - General information and its regulations --- p.11 / Chapter 1.2.2. --- Cortisol - Functions --- p.12 / Chapter 1.3. --- Thyroid hormones --- p.14 / Chapter 1.3.1. --- THs - General information and its regulations --- p.14 / Chapter 1.3.2. --- THs - Functions --- p.15 / Chapter 1.4. --- Growth hormone --- p.18 / Chapter 1.4.1. --- GH - General information and its regulations --- p.18 / Chapter 1.4.2. --- GH - Functions --- p.20 / Chapter 1.5. --- Insulin-like growth factor --- p.22 / Chapter 1.5.1. --- IGF-I - General information and its regulations --- p.22 / Chapter 1.5.2. --- IGF-I - Functions --- p.24 / Chapter 1.6 --- Metallothioneins --- p.26 / Chapter 1.6.1. --- MTs - Definition and Classification --- p.26 / Chapter 1.6.2. --- MTs - Functions --- p.27 / Chapter 1.7. --- Glucose-6-phosphate dehydrogenase --- p.31 / Chapter 1.7.1. --- G6PDH - General information and its regulations --- p.31 / Chapter 1.7.2. --- G6PDH ´ؤ Functions --- p.32 / Chapter Chapter 2: --- "Effects of cadmium exposure on the endocrine status of silver sea bream, Sparus sarba" --- p.34 / Chapter 2.1. --- Introduction --- p.35 / Chapter 2.2. --- Materials and methods --- p.37 / Chapter 2.2.1. --- Overall experimental design --- p.37 / Chapter 2.2.2. --- In vivo exposure to waterborne cadmium --- p.37 / Chapter 2.2.2.1. --- Experimental animals --- p.37 / Chapter 2.2.2.2. --- Adaptation --- p.37 / Chapter 2.2.2.3. --- Tissue sampling --- p.38 / Chapter 2.2.2.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.38 / Chapter 2.2.2.4.1. --- Serum cortisol analysis --- p.39 / Chapter 2.2.2.4.2. --- Serum triiodothyronine analysis --- p.39 / Chapter 2.2.2.4.3. --- Serum thyroxine analysis --- p.39 / Chapter 2.2.2.5. --- Protein extraction and Protein quantification --- p.40 / Chapter 2.2.2.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.40 / Chapter 2.2.2.7. --- RNA extraction --- p.41 / Chapter 2.2.2.8. --- Reverse transcription for first-strand cDNAs from total RNAs samples from liver --- p.42 / Chapter 2.2.2.9. --- Real-time quantitative PCR assays of IGF-I mRNA expression --- p.43 / Chapter 2.2.3. --- In vivo experiments involving cadmium injection --- p.44 / Chapter 2.2.3.1. --- Experimental animals --- p.44 / Chapter 2.2.3.2. --- Adaptation --- p.44 / Chapter 2.2.3.3. --- Tissue sampling --- p.45 / Chapter 2.2.3.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.45 / Chapter 2.2.3.4.1. --- Serum cortisol analysis --- p.45 / Chapter 2.2.3.4.2. --- Serum triiodothyronine analysis --- p.45 / Chapter 2.2.3.4.3. --- Serum thyroxine analysis --- p.46 / Chapter 2.2.3.5. --- Protein extraction and Protein quantification --- p.46 / Chapter 2.2.3.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.46 / Chapter 2.2.3.7. --- RNA extraction --- p.46 / Chapter 2.2.3.8. --- Reverse transcription for the first-strand cDNAs from total RNAs samples from liver --- p.46 / Chapter 2.2.3.9. --- Real-time quantitative PCR of IGF-I mRNA expression --- p.46 / Chapter 2.2.4. --- In vitro part of the project (Primary cell culture: hepatocytes exposed to cadmium medium) --- p.47 / Chapter 2.2.4.1. --- Experimental animals --- p.47 / Chapter 2.2.4.2. --- Primary hepatocytes culture preparation --- p.47 / Chapter 2.2.4.3. --- Cadmium treatment and cell harvest --- p.48 / Chapter 2.2.4.4. --- "RNA extraction, reverse transcription for the first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of IGF-I mRNA expression" --- p.48 / Chapter 2.2.5. --- Statistical analysis --- p.48 / Chapter 2.3. --- Results --- p.49 / Chapter 2.3.1. --- Serum cortisol level --- p.49 / Chapter 2.3.2. --- Serum triiodothyronine level --- p.49 / Chapter 2.3.3. --- Serum thyroxine level --- p.49 / Chapter 2.3.4. --- Pituitary growth hormone level --- p.50 / Chapter 2.3.5. --- Hepatic insulin-like growth factor mRNA expression --- p.50 / Chapter 2.4. --- Discussion --- p.58 / Chapter 2.4.1. --- Serum cortisol level --- p.58 / Chapter 2.4.2. --- Thyroid hormones --- p.61 / Chapter 2.4.3. --- Growth hormone --- p.64 / Chapter 2.4.4. --- Insulin-like growth factor-I --- p.67 / Chapter 2.5. --- Conclusion --- p.70 / Chapter Chapter 3: --- "Effects of cadmium exposure on MT and G6PDH mRNA expressions of silver sea bream, Sparus sarba" --- p.71 / Chapter 3.1. --- Introduction --- p.72 / Chapter 3.2. --- Materials and methods --- p.74 / Chapter 3.2.1. --- Overall experimental design --- p.74 / Chapter 3.2.2. --- In vivo experiments involving exposure to waterborne cadmium --- p.74 / Chapter 3.2.2.1. --- Experimental animals --- p.74 / Chapter 3.2.2.2. --- Adaptation --- p.74 / Chapter 3.2.2.3. --- Tissue sampling --- p.74 / Chapter 3.2.2.4. --- RNA extraction --- p.74 / Chapter 3.2.2.5. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.75 / Chapter 3.2.2.6. --- Amplification of partial fragments of metallothionein (MT) --- p.75 / Chapter 3.2.2.7. --- Rapid amplification of 5´ة and 3´ة cDNA ends of metallothionein (MT) --- p.76 / Chapter 3.2.2.7.1. --- Amplification of 5´ة cDNA end --- p.76 / Chapter 3.2.2.7.2. --- Amplification of 3´ة cDNA end --- p.78 / Chapter 3.2.2.8. --- Real-time quantitative PCR of MT and G6PDH mRNA expressions --- p.79 / Chapter 3.2.3. --- In vivo injection of cadmium --- p.80 / Chapter 3.2.3.1. --- "Experimental animals, adaptation and tissue sampling" --- p.80 / Chapter 3.2.3.2. --- RNA extraction --- p.80 / Chapter 3.2.3.3. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.81 / Chapter 3.2.3.4. --- Real-time quantitative PCR of MT and G6PDH mRNA expression --- p.81 / Chapter 3.2.4. --- In vitro exposure of primary hepatocyte culture to cadmium --- p.81 / Chapter 3.2.4.1. --- Experimental animals --- p.81 / Chapter 3.2.4.2. --- Preparation of the hepatocytes for cell culture --- p.81 / Chapter 3.2.4.3. --- Cadmium treatment and cell harvest --- p.81 / Chapter 3.2.4.4. --- "RNA extraction, reverse transcription for first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of MT and G6PDH mRNA expression" --- p.82 / Chapter 3.2.5. --- Statistical analysis --- p.82 / Chapter 3.3. --- Results --- p.83 / Chapter 3.3.1. --- MT cloning and characterization --- p.83 / Chapter 3.3.2. --- Metallothionein mRNA expression --- p.83 / Chapter 3.3.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.84 / Chapter 3.4. --- Discussion --- p.91 / Chapter 3.4.1. --- MT cloning and characterization --- p.91 / Chapter 3.4.2. --- Metallothioneins mRNA expression --- p.92 / Chapter 3.4.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.95 / Chapter 3.5. --- Conclusion --- p.98 / General conclusion --- p.99 / References --- p.101

Diabetes and insulin resistance are on the rise in the United States. Early detection and deployment of therapies has allowed for the reversal of pancreatic beta cell damage. Unfortunately, not all providers can offer the support to facilitating the required life style modifications. The introduction of clinical health consultants (CHC) as supplemental care has improved patient health for a variety of chronic diseases. Missing in the literature are studies investigating the correlation between the number of CHC interactions and improvement in biomarkers. The study utilized a non-experimental, retrospective study design to evaluate the relationship between the use between the use of CHCs and the number of CHC interactions, and the mean changes in glucose, hemoglobin A1c, insulin, proinsulin, C-peptide, and 1,5-anhydroglucitol, over a one-year period for patients presented with the opportunity to participate in CHC interactions. The subjects’ follow-up results were compared to their initial results for each group using the ANCOVA and one-way t-test. A statistically significant difference was detected between the mean change in BMI and the use of CHCs (p

[Truncated abstract] Every year thousands of individuals suffer from thrombotic related complications that in some cases can be fatal and every year millions of women take some form of hormonal contraceptive. In some cases, there is a cause and effect relationship between the two as users of the combined oral contraceptive pill have an increased risk of developing a thrombotic event. Increased circulating levels of oestrogen cause a prothrombotic shift in the coagulation cascade resulting from upregulation of several procoagulant proteins and a decrease of key anticoagulant proteins. One of the most oestrogen sensitive anticoagulants is Protein S (PS), a product of the PROS1 gene. PS acts as a cofactor to activated protein C (aPC) and the PS-aPC complex serves to downregulate clot formation by deactivating the tenase and prothrombinase complexes via proteolytic cleavage of activated factors VIII and V, respectively. As such, low PS levels are associated with an increased risk of developing thrombotic disorders such as pulmonary embolism, stroke or coronary thrombosis and deep vein thrombosis. During pregnancy when oestrogen levels increase, a steady decline in PS is evident in the early weeks of gestation and continues to decrease to below the normal range in the 2nd trimester, remaining there until post-partum. In addition, reduced free and total PS levels are observed in users of the combined oral contraceptive (COC) pill that contains an oestrogen and a progestin. Interestingly, users of 3rd generation COCs have significantly greater reductions of PS than do 2nd generation COC users. The difference between the two forms is the type of progestin, not the oestrogen, which is predominantly ethinyl oestradiol in the majority of commercially available preparations. At present, a mechanism to describe the relationship between oestrogen and/or progesterone associated with the observed in vivo changes in the levels of PS has not been identified. The aim of this thesis was to define the molecular mechanisms involved in the regulation of PS expression by oestrogen and progesterone. In this study, a Combined Single-stranded conformational analysis and Heteroduplex Analysis (CSHA) iv methodology was optimised for screening both PROS1 DNA and mRNA for the detection of mutations. '...' This may explain why users of 3rd generation COCs display a greater reduction in circulating PS levels compared to 2nd generation users. To investigate potential PS interactions with other proteins that could be hormonally regulated, a yeast-2-hybrid (Y-2-H) screen was performed using the PS molecule as a 'bait' against molecules derived from liver and bone marrow cDNA libraries. A clone that contained a portion of another haemostatic protein, Protein Z (PZ) was isolated and confirmed via sequencing. As no full length PZ clones were identified, a second Y-2-H screen was performed once again using the PS molecule as bait and the PZ molecule as the fish. Interaction between the two proteins was shown to be possible via the successful growth of colonies on triple knock out selective media and by positive ß-galactosidase activity.

No description available.

<p>Version:1.0 StartHTML:0000000178 EndHTML:0000005278 StartFragment:0000002640 EndFragment:0000005242 SourceURL:file://localhost/Volumes/NAMNLOS/Examensarbete%20kortisol.doc</p><p>Föreliggande studie syftar till att försöka utläsa skillnader mellan två olika perioder då den personliga stressfaktorn upplevs vara olika intensiv. Undersökningen syftar även till att studera huruvida den mänskliga kortisolutsöndringens diurnala upp - och ned gångar följer en intraindividuell stabilitet av CAR (cortisol awakening responce). Detta skulle innebära ett upprepande mönster av kortisolkoncentrationens magnitud och mätvärde inom varje individ från dag till dag, vid uppvaknandet och 30 minuter efter.</p><p>Undersökningen har genomförts som en pilotstudie där en försökspersons kortisolkoncentration i saliv har mätts genom enzymkopplad immunabsorberande analys (ELISA). För att jämföra mätserierna inom de olika perioderna med varandra har även en variationsanalys av typen Analysis of variance (ANOVA) utförts med hjälp av programvaran SPSS. Då provernas mätvärde har analyserats och jämförts med varandra har ett resultat kunnat fastställas.</p><p>Eftersom utsöndringen av den individuella kortisolkoncentrationen lätt påverkas av omgivningsfaktorer användes endast en försöksperson, författaren, vilket underlättade en detaljerad analys där observation av påverkande faktorer lätt kunde tas med i beräkningen för att fastställa ett tillförlitligt resultat. Försökspersonen, kvinna 21 år, utförde 6 provtagningar under två perioder som upplevdes ha olika hög stressfaktor. Perioderna innehöll två arbetsdagar. Parallellt med provtagningen fördes noggranna dagboksanteckningar för att underlätta analyseringsarbetet.</p><p>Resultatet uppvisar en intraindividuell stabilitet av CAR hos försökspersonen. Studien visar även en skillnad mellan de två perioderna genom en högre procentuell ökning av CAR under den period då stressfaktorn upplevdes som mer intensiv.</p><p>Den tydliga skillnaden av kortisolkoncentrationens mätvärde mellan de olika dagarna indikerar även att livsstil, fysisk aktivitet och drömmar kan påverka utseendet av kortisolkoncentrationskurvans diurnala upp – och nedgångar.</p>

The work presented in this dissertation examines possible modes of action for growth inhibition by anthropogenic endocrine disrupting chemicals (EDCs) as well as endogenous hormones associated with growth in fish. Using the sheepshead minnow (SHM) (Cyprinodon variegatus) as a model, I developed methods to examine perturbations in the endocrine axis controlling fish growth, and also examined effects of EDCs on the whole fish. I used two relatively new techniques to study the endocrine growth axis, quantitative real-time PCR (TaqMan) and differential display analysis. TaqMan analysis is a highly sensitive method to measure specific sequences from a small amount of total RNA using a fluorescent probe and specific primer pairs. I optimized a TaqMan assay for SHM IGF-I to measure hepatic IGF-I mRNA concentrations. in fish injected with hormones known to influence fish growth (GH, T���, E���, insulin, or a carrier control). IGF-I mRNA levels increased in fish injected with GH, T��� and insulin, peaking at 12 h post-injection. IGF-I mRNA levels decreased significantly at 8 h and 12 h post-injection in fish injected with E���, suggesting that pharmacological levels of E��� may affect the GH/IGF-I axis and could have consequences for fish living in waters polluted by EDCs. Differences in growth were observed in fish exposed for 18 weeks to E��� or chlorpyrifos (an organophsophate). Fish exposed to the highest dose of E��� grew larger than controls only during the last week of the experiment. Fish exposed to the lower dose of E��� were not significantly different from controls. The fish exposed to all doses of chloryprifos grew significantly less than controls in a dose-dependent manner. No significant differences were found in hepatic IGF-I mRNA levels in any treatments. To establish patterns of gene up- or down-regulation, I performed differential display analysis on livers of several fish from the previous two experiments. Several genes were identified as being similar to fish including a microsatellite sequence, a choriogenin (vitelline envelope) protein mRNA sequence, a transferrin mRNA sequence and several ribosomal RNA sequences. This technique to evaluate gene expression will become more useful when more fish genes are added to the data bases. / Graduation date: 2003

The global population of leatherback sea turtles is decreasing worldwide, with extinction predicted for some populations within 15 years. The population of leatherbacks nesting at Sandy Point National Wildlife Refuge (SPNWR), St. Croix, USVI, displayed a significant population increase from 1982 2001 but has experienced a slowed recovery since then. To better understand the causes of this decline, a historical database of SPNWR nesting female data was utilized to investigate trends in reproductive indices. Since 2001, average remigration interval (RI) has increased significantly, while average number of clutches laid, hatch success, hatchling production, and the percentage of neophytes recruited annually have decreased. Annual remigrant numbers have been stable to increasing, suggesting that adult survivorship remains high. To assess whether maternallyderived factors may be influencing clutch production and low hatch success, blood samples were collected by saturation sampling during nesting. Circulating estradiol, testosterone, and progesterone were evaluated in conjunction with reproductive data. All hormones were highest at deposition of the first clutch and declined progressively with each consecutive clutch, as previously observed in other sea turtle species. Increased clutch production in remigrants was associated with higher estradiol levels compared to neophytes, presumably due to ovarian size and maturity. Contrary to observations in Pacific leatherbacks, progesterone decreased significantly with successive nests and total levels of estrogen were significantly lower, suggesting Atlantic leatherbacks may undergo a longer migration or spend more time in the feeding grounds prior to migrating. Linear Mixed Effect (LME) modeling was employed to determine whether hormone levels at nesting might serve as indicators of reproductive variables. Because models for all hormones were individual specific, a population model could not be developed that effectively utilized hormone levels at nesting to predict clutch size, hatch success, age or RI. However, number of clutches laid may potentially be predicted based on individually tailored estrogen models. Decreased recruitment (due to increased mortality of early life stages, altered sex ratios, or delayed age to sexual maturity), decreased productivity, and increased RI (possibly due to diminished foraging ground productivity) appear primarily responsible for current population trends which threaten the population's future.

<p>Female social dominance over males is unusual in mammals, yet characterizes most Malagasy lemurs, which represent almost 30% of all primates. Despite its prevalence in this suborder, both the evolutionary trajectory and proximate mechanism of female dominance remain unclear. Potentially associated with female dominance is a suite of behavioral, physiological and morphological traits in females that implicates ‘masculinization’ via androgen exposure; however, relative to conspecific males, female lemurs curiously show little evidence of raised androgen concentrations. In order to illuminate the proximate mechanisms underlying female dominance in lemurs, I observed mixed‐sex pairs of related Eulemur species, and identified two key study groups ‐‐ one comprised of species expressing female dominance and, the other comprised of species (from a recently evolved clade) showing equal status between the sexes (hereafter ‘egalitarian’). Comparing females from these two groups, to test the hypothesis that female dominance is an expression of an overall masculinization of the female, I 1) characterize the expression of female dominance, aggression, affiliation, and olfactory communication in Eulemur; 2) provide novel information about the hormonal and neuroendocrine correlates associated with the expression of female dominance; 3) investigate the activational role of the sex-steroid hormones in adult female Eulemur using seasonal correlates of hormonal and behavioral change; and 4) examine the specific role of estrogen in the regulation and expression of sex-reversed female behavior in these species. In doing so I highlight significant behavioral and physiological differences between female-dominant and egalitarian Eulemur and show that female dominance is associated with a more masculine behavioral and hormonal profile. I also suggest that these behavioral and hormonal differences may be the result of fundamental differences in the biosynthetic pathway associated with estrogen production. Moreover, I assert that these putative physiological differences could provide a parsimonious proximate mechanism explaining the evolution of female dominance and its subsequent relaxation in egalitarian Eulemur species.</p> / Dissertation

Σκοπός της μελέτης ήταν η διερεύνηση της σχέσης ανάμεσα στο μεταβολισμό του σιδήρου και το σακχαρώδη διαβήτη τύπου 2. Έτσι ο στόχος της παρούσης μελέτης ήταν διπλός. α) Να καθορισθεί η συχνότητα των μεταλλάξεων C282Y και H63D στον Ελληνικό πληθυσμό και να συγκριθεί με αυτήν άλλων χωρών και ταυτοχρόνως, να ελεγχθεί αν η συχνότητα των ανωτέρω μεταλλαγών είναι διαφορετική στους διαβητικούς τύπου 2 σε σχέση με το γενικό πληθυσμό και να γίνει συσχέτιση ανάμεσα στην ύπαρξη η όχι των μεταλλάξεων αυτών και σε δείκτες μεταβολισμού του σιδήρου όπως η φερριτίνη, ο σίδηρος, ο κορεσμός τρανσφερίνης. β) Να ελεγχθεί αν στους διαβητικούς τύπου 2, η συχνότητα της κληρονομικής αιμοχρωμάτωσης (ομοζυγώτες), όπως αυτή μπορεί να καθορισθεί με βάση το γονότυπο του γονιδίου HFE είναι αυξημένη σε σχέση με το γενικό πληθυσμό. Υλικό και μέθοδοι Αρχικά δείγμα 100 διαβητικών τύπου 2 και 100 ατόμων από το γενικό πληθυσμό ελέγθησαν για τις μεταλλάξεις του γονιδίου HFE C282Y και H63D με τη μέθοδο της PCR και RFLP. Στη συνέχεια μελετήθηκαν 500 διαβητικοί τύπου 2 και 423 μάρτυρες. Στα άτομα αυτά εκτιμήθηκε το φορτίο του οργανισμού σε σίδηρο (Σίδηρος ορού, ολική δεσμευτική ικανότητα σε σίδηρο, φερριτίνη) και καθορίστηκε ο γονότυπος αναφορικά με τις μεταλλάξεις C282Y και H63D με τη μέθοδο της PCR και RFLP. Αποτελέσματα- Συμπεράσματα 1. Η συχνότητα του αλληλομόρφου C282Y στον Ελληνικό πληθυσμό(0,0075) είναι χαμηλότερη των ατόμων Βορειοευρωπαικής καταγωγής και πλησιάζει αυτή ατόμων από περιοχές της νότιας Ευρώπης. Η συχνότητα του αλληλομόρφου H63D (0,115) είναι παρόμοια με αλλους πληθυσμούς. 2. Δεν ανευρέθη διαφορά στη συχνότητα των μεταλλάξεων C282Y και H63D ανάμεσα στην ομάδα των διαβητικών τύπου 2 και στο γενικό πληθυσμό. 3. Η παρουσία οποιασδήποτε από τις παραπάνω μεταλλάξεις στην ετερόζυγη μορφή συνεισφέρει στην αύξηση του φορτίου του οργανισμού σε σίδηρο. Αυτό βρέθηκε και στις δύο ομάδες. 4. Οι διαβητικοί τύπου 2 παρουσιάζουν μεγαλύτερο κορεσμό τρανσφερρίνης αίματος και υψηλότερα επίπεδα φερριτίνης συγκρινόμενοι με το γενικό πληθυσμό. Οι ανωτέρω δείκτες θα μπορούσαν να αντιπροσωπεύουν αυξημένο φορτίο του οργανισμού σε σίδηρο. 5. Λόγω της χαμηλής συχνότητας του αλληλομόρφου C282Y δεν κατέστη δυνατή η ανίχνευση ομοζυγωτών C282Y/ C282Y. / Several authors have suggested a positive association between diabetes type 2 and the C282Y and H63D mutations of the hereditary hemochromatosis gene but others have disputed it. There are also papers reporting an increased iron load in diabetes type 2 and possible associations with the pathogenesis of the disease. We performed therefore a study in 100 type 2 diabetics and 100 age and sex matched controls to assess the role of the C282Y and H63D mutations in the HFE gene as a risk factor for type 2 diabetes mellitus in Greece. We also evaluated the iron load in 500 diabetes type 2 patients and 423 age and sex matched controls. We did not find any differences in the allele frequencies of the above mutations between patients with diabetes type 2 and the controls. The allele frequencies are estimated to be 0.0075 for the C282Y and 0.115 for the H63D mutation. Subjects with at least one mutation (C282Y or H63D) had higher transferrin saturation compared to those with no such mutations. This seems to apply to both diabetics (49± 8,6 vs 44,5± 5,4, p<0,01) and control group (49,3± 7,3 vs 42,6± 3,3 p<0,01). Patients with diabetes type 2 have higher transferrin saturation compared to the general population. These differences were found among men (n=250, mean± SD 31,8+11 vs n=73, mean± SD 29,5+8, p=0,05) as well as among women (n=250, mean± SD 28.5+10 vs n=350, mean± SD 25.5+9.6, p=0.001). The present study is in concord with previous work reporting that type 2 diabetes patients have higher ferritin levels compared to controls.

The objective of this dissertation research was to determine the specific actions of chronically elevated catecholamines on; 1.) fetal growth and ß-cell function during the third trimester in vivo in an ovine model of placental insufficiency-induced intrauterine growth restriction (PI-IUGR), and 2.) regulation of insulin secretion in vitro utilizing the mouse insulinoma cell line Min6.At 0.7-gestation, fetal weights were not different but PI fetuses had lower (P<0.05) basal blood oxygen content, plasma glucose, IGF-1, and insulin concentrations and greater norepinephrine concentrations (891±211 vs. 292±65 pg/ml; P<0.05) compared to controls. Glucose-stimulated insulin secretion (GSIS) was lower in PI than control fetuses (0.34±0.03 vs. 1.08±0.06 ng/ml; P<0.05). ADR-block increased GSIS in PI fetuses (1.19±0.11) but decreased GSIS in controls (0.86±0.02 ng/ml). Insulin content per islet was not different between PI and control fetuses. We concluded that elevated fetal plasma norepinephrine, in PI fetuses at 0.7 gestation, precedes growth restriction and suppresses insulin concentrations, and ADR-block revealed compensatory β-cells stimulus-secretion responsiveness. Therefore, to determine the effects of chronic hypercatecholamine exposure on fetal growth and β-cell function independent of hypoglycemia and hypoxemia, we performed surgical sham or adrenal demedullation (AD) at 0.65 gestation on control and IUGR fetuses (n= 5 Control-Sham, 5 Control-AD, 5 IUGR-Sham, 5 IUGR-AD fetuses). Studies commenced at 0.9 gestation under ambient conditions and steady-state reversal of arterial pO2 between IUGR and control fetuses. Plasma norepinephrine was 5-fold higher in IUGR-Sham vs. Control-Sham and reduced in IUGR-AD fetuses to concentrations not different from Control-Sham fetuses. Fetal mass was lower in IUGR vs. control fetuses but 92% greater in IUGR-AD compared to IUGR-Sham fetuses. Basal plasma glucose and arterial pO2 were lower in IUGR-Sham vs. Control-Sham, and IUGR-AD vs. Control-AD fetuses. Basal and glucose-stimulated insulin concentrations compared to Control-Sham were lower in IUGR-Sham and IUGR-AD and Control-AD fetuses. Oxygenation improved GSIS in IUGR-Sham and IUGR-AD fetuses. In conclusion, hypoglycemia, hypoxemia and norepinephrine interdependently and differentially regulate aspects of fetal growth and β-cell function in the IUGR fetus. In Min6 cells, we determined that GSIS responsiveness is enhanced and adrenergic receptor α2A is desensitized cells following chronic exposure to epinephrine.