Dissertations / Theses on the topic 'Megalina'

Introdução: O papel do sistema HLA na evolução do transplante é indiscutível. Dados da literatura internacional e do nosso laboratório têm mostrado que este não é o único sistema envolvido nos processos de RMA (rejeição mediada por anticorpo) Esse fato é comprovado a partir da constatação de que transplantes realizados entre irmãos com total identidade HLA também são alvos da RMA. Entretanto, os alvos antigênicos desses anticorpos permanecem desconhecidos, dificultando assim o diagnostico e tratamento da RMA não-HLA. No transplante renal a presença desses anticorpos têm sido associada com anticorpos anti células endoteliais, células epiteliais tubulares, podocitos, células mesangiais e monócitos. Nosso objetivo neste estudo foi avaliar a presença e a relevância clínica de anticorpos contra a megalina, membro da família de receptores de LDL, expressa na membrana apical dos túbulos proximais, com importante papel na reabsorção de proteínas no rim. Métodos: Soros pré-transplante de 105 pacientes submetidos a transplante renal, realizado no Serviço de Transplante Renal do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (UTR-HC-FMUSP) foram testados por ELISA contra 2 peptídeos da megalina estudados e sintetizados em nosso laboratório. (convencionalmente chamados 18 e 19) Resultado: Não foi detectada a presença de anticorpos do isotipo IgG nas amostras prétransplante dos pacientes estudados. Entretanto, foi detectada a presença de anticorpos do isotipo IgM anti-peptídeo 18 em 33 (31,4%) amostras de soro e anti-peptídeo 19 em 23 (21,9%). Para avaliar a significância clinica desses anticorpos dividimos os pacientes em 2 grupos: pacientes com pelo menos 1 episodio de rejeição aguda (Grupo I) e pacientes sem rejeição (Grupo II) e observamos a distribuição dos pacientes positivos nos 2 grupos. O numero de pacientes com anticorpos anti-peptídeo 18 foi igualmente distribuída nos dois grupos. (12/42, 28,5% no Grupo I e 21/63, 33,3% no Grupo II p=ns). A maioria dos pacientes com anticorpos anti-peptídeo 19 pertenciam ao grupo I (17/42, 40,5% ; 6/63, 9,5% p=0,0003). Esta analise demonstrou uma boa correlação entre presença de anticorpos anti-peptídeo 19 no soro pré-transplante e rejeição. Conclusão: Nossos dados sugerem que anticorpos IgM anti-megalina no pré-transplante podem ser um fator de risco na rejeição do aloenxerto renal. É importante salientar que a nosso conhecimento este é o primeiro estudo envolvendo megalina e rejeição no transplante clínico.
Background. Preformed donor-specific human leukocyte antigen (HLA)- antibodies are accountable for the majority of antibody-mediated rejections (AMR). However, recipients of HLA identical kidneys can develop AMR implicating putative pathogenic antibodies that are directed against non-HLA antigens. Unknown immune targets and consecutive lack of detection methods make non-HLA AMR particularly difficult to diagnose and treat. In renal transplant rejection, the presence of antibodies to non-HLA has been associated with antibodies against endothelial cells, tubular epithelial cells, podocytes, mesangial cells and monocytes. The aim of this study was to evaluate the presence and clinical relevance of preformed antibodies against megalin, a member of the LDL receptor family, expressed on the apical membrane of proximal tubules. Megalin performs a central role in renal protein reabsorption. Methods. Pré-transplant sera of 105 recipients of kidney allograft transplanted at Serviço de Transplante Renal do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (STRHCFMUSP) were tested by enzyme-linked immunosorbent assay (ELISA) against 2 megalin-peptides (conventionally named 18 and 19) studied and synthetized in our laboratory. Results. Antibodies were not detected in pretranplant sera when tested for IgG isotype. However, in 33 (31,4%) sera we detected the presence of IgM antibodies to megalin-peptide18 and in 23 (21,9%) to megalin-peptide 19. To evaluate the clinical significance of these antibodies we divided the patients in 2 groups: Group I - 42 patients with at least one rejection episode during the first post-transplant year and Group II - 63 patients without any rejection episode and observed the distribution of positive patients in each of the 2 groups. Patients with megalin-peptide18 antibodies had the same distribution in both groups. (12/42, 28, 5% Group I and 21/63, 33,3% no Group II p=ns). However, patients with megalinpeptide19 antibodies were more frequent in group I. (17/42, 40,5% group I and 6/63, 9,5% group II p=0,0003). This analysis demonstrated a good correlation between preformed anti-megalin-peptide19 antibodies and allograft rejection Conclusion. Our data suggest that presence of IgM megalin antibodies before transplantation might be a risk factor for kidney allograft rejection. To our knowledge, this is the first study involving megalin and rejection in clinical transplantation.

Evidências epidemiológicas indicam que a presença de microalbuminúria prediz maior freqüência de eventos cardiovasculares e mortalidade em hipertensos essenciais. A microalbuminúria pode ser decorrente do aumento da permeabilidade glomerular e/ou da diminuição da reabsorção desta macromolécula no túbulo proximal. Todavia não é sabido se os mecanismos que regulam a reabsorção de albumina em túbulo proximal renal encontram-se alterados na hipertensão essencial. Este trabalho teve como objetivo investigar as bases moleculares da microalbuminúria associada à hipertensão arterial essencial, focando na reabsorção tubular de albumina. Para tanto, avaliamos a evolução temporal da excreção urinária de albumina em ratos espontaneamente hipertensos (SHR) com 6 semanas de idade (pressão arterial sistólica, PAS, = 105 ± 4 mmHg), 14 semanas de idade (PAS = 180 ± 2 mmHg) e 21 semanas de idade (PAS = 202 ± 2 mmHg). Ratos normotensos Wistar da mesma idade serviram de controle. Observou-se que a excreção urinária diária de albumina aumentou progressivamente com o aumento da pressão arterial em SHR (10,5 ± 1,9; 92 ± 7,0 e 154 ± 27 g/dia, em SHR com PAS média igual a 105, 180 e 202 mmHg respectivamente). Este aumento progressivo não foi observado em ratos normotensos com idade correspondente, indicando que este fenômeno é decorrente do aumento da pressão arterial e não pode ser atribuído ao aumento da idade dos animais durante o período estudado. A análise das proteínas urinárias por eletroforese em gel de poliacrilamida (SDS-PAGE) mostrou que SHR excretam proteínas do tamanho da albumina ou menores (< 70kDa), padrão típico de proteinúria tubular. Adicionalmente, verificou-se que os níveis de expressão dos receptores endocíticos megalina e cubilina, bem como do canal para cloreto ClC-5 diminuem progressivamente no córtex renal de SHR com o aumento da pressão arterial. Observou-se também uma diminuição significativa na expressão de uma outra macromolécula importante no processo de endocitose mediada por receptor em túbulo proximal renal, a v-H+-ATPase. Entretanto, a diminuição da expressão protéica da subunidade B2 desta ATPase foi estatisticamente significante apenas em SHR com 21 semanas comparado aos com 6 semanas de idade. Não foram encontradas alterações no padrão de expressão de componentes estruturais da barreira glomerular como a nefrina e podocina. Em suma, o nosso estudo demonstra que o aumento da excreção urinária de proteínas, especialmente de albumina, está associado com uma menor expressão de componentes essenciais do aparelho endocítico do túbulo proximal renal. É tentador especular que a disfunção da via endocítica no túbulo proximal renal possa ser o principal mecanismo subjacente ao desenvolvimento de microalbuminúria na hipertensão
Epidemiological evidences indicate that the presence of microalbuminuria predicts a higher frequency of cardiovascular events and mortality in essential hypertensive patients. Microalbuminuria may arise from increased glomerular permeability and/or reduced proximal tubular reabsorption of albumin. However, it remains to be determined whether the mechanisms that regulate the renal proximal tubular reabsorption of albumin are altered in essential hypertension. The purpose of this work was to investigate the molecular basis of microalbuminuria in essential hypertension, focusing on the renal tubular reabsorption of albumin. To this end, we evaluated the temporal evolution of urinary albumin excretion in spontaneously hypertensive rats (SHR) at 6 weeks of age (systolic arterial pressure, SAP, = 105 ± 4 mmHg), 14 weeks of age (SAP = 180 ± 2 mmHg) and 21 weeks of age (SAP = 202 ± 2 mmHg). Age-matched normotensive Wistar rats were used as controls. It was observed that the daily urinary excretion of albumin progressively increased with blood pressure in SHR from 6 to 21 weeks of age (10.5 ± 1.9, 92 ± 7.0 and 154 ± 27 g in SHR with 105, 180 and 202 mmHg of average SAP, respectively). This progressive increase in microalbuminuria has not been observed in age-matched normotensive Wistar rats, indicating that this phenomenon cannot be attributed to age progression over the studied period. SDS-PAGE analysis of urinary proteins showed that microalbuminuric SHR virtually excreted proteins of the size of albumin or smaller (< 70kDa), typical of tubular proteinuria. Additionally, it was verified that the protein expression levels of the endocytic receptors megalin and cubilin as well as of the chloride channel ClC-5 progressively decreased in the renal cortex of SHR from 6 to 21 weeks of age. Moreover, it was observed reduction of expression of another macromolecule that plays an important role in the process of receptor mediated endocytosis in the renal proximal tubule, the v-H+- ATPase, was reduced. However, reduced cortical expression of the B2 subunit of the v- H+-ATPase, was only statistically significant in 21-wk-old vs. 6-wk-old SHR. Expression levels of structural components of the glomerular barrier such as nephrin and podocin were unchanged. To sum up, our study demonstrates that the increase in urinary protein excretion, especially of albumin, is associated with lower expression of key components of the apical endocytic apparatus in the renal proximal tubule. It is tempting to speculate that dysfunction of the apical endocytic pathway in the renal proximal tubule may be the major mechanism underlying development of microalbuminuria in essential hypertension

ClC-5 and Megalin are part of the same macromolecular complex involved in the re-uptake of albumin in the proximal tubuli. To understand the relationship between this mechanism and proteinuric kidney diseases, we have performed gene expression study of CLCN5 and Megalin genes in glomerular (gl) and tubular interstitial (ti) compartments of patients with type 2 diabetes (NIDDM) and IgA nephropathy (IgAN). Gene expression of CLCN5 and Megalin was investigated by Real-Time PCR in microdissected biopsies (mpb) (under a stereomicroscope) from patients with IgAN (n 10) and with NIDDM (n 9). Both groups had similar level of proteinuria. As controls, we used cortical tissues obtained from sites remote from tumor bearing renal tissue (n 9). Moreover, we analyzed human mesangial cells (HMC), endothelial cells (HuVEC), tubular cells (HK-2) and podocytes (HP) in basal conditions. HMC and HK-2 were also examined in high glucose and albumin conditions respectively. We found that: 1) CLCN5 and Megalin were expressed in all mbp both at gl and ti levels, and their expression did not differ between gl and ti of NIDDM and IgAN; 2) CLCN5 and Megalin expression levels were significantly higher in proteinuric nephropathies than in controls both at gl and ti compartment. In order to exclude ti contamination in microdissected glomeruli, we performed gene expression analysis of CLCN5 and Megalin also on laser microdissected glomeruli from 28 NIDDM patients and 14 controls that confirmed our previous results. In these mbp a direct correlation between CLCN5 and Megalin was evidenced (r=0,50 p=0,0039); 3) CLCN5 transcript was evidenced in all cells analyzed, on the contrary, very low levels of Megalin were detected; 4) HK-2 treated with albumin showed the same level of CLCN5 expression as untreated cells; 5) HMC treated with high glucose for 24 and 48 hours showed the same level of CLCN5 expression as untreated cells. Our study reveals, for the first time, that CLCN5 and Megalin genes are expressed in human glomeruli, and that all the cell types constituting the glomerulus express in vitro CLCN5. The overexpression of CLCN5 and Megalin in diabetic and IgA nephropathy biopsies suggests that proteinuria might regulate their expression. The in vitro experiments on HMC exclude the role of glucose in the up-regulation of ClC-5 expression in diabetic glomerulopathy. A role might be hypothesized for podocytes since podocyte endocytosis of proteins does occur in proteinuric patients.
ClC-5 e Megalina sono due delle proteine che compongono il complesso macromolecolare coinvolto nel riassorbimento dell’albumina a livello del tubulo prossimale. Per comprendere meglio se ci fosse una relazione tra questo meccanismo e le patologie proteinuriche abbiamo effettuato uno studio di espressione per i geni CLCN5 e Megalina nei compartimenti glomerulare e tubulo interstiziale di pazienti con diabete di tipo II (NIDDM) e nefropatia a depositi di IgA (IgAN). L’espressione di CLCN5 e Megalina è stata investigata mediante Real-Time PCR in biopsie microdissezionate manualmente grazie all’ausilio di uno stereomicroscopio. Queste sono stete ottenute da pazienti con diabete di tipo II (n°9) e nefropatia a depositi di IgA (n°9). Come controllo è stato utilizzato tessuto corticale ottenuto da polo renale indenne in corso di nefrectomia per tumore. Entrambi i gruppi di pazienti analizzati presentavano livelli simili di proteinuria. Abbiamo inoltre analizzato cellule mesangiali umane (HMC), cellule endoteliali umane (HuVEC) cellule tubulari umane (HK-2) e podociti umani (HP) in condizioni basali. HMC e HK-2 sono state anche analizzate rispettivamente in presenza di alte concentrazioni di glucosio e a diverse concentrazioni di albumina. Abbiamo trovato che: 1) CLCN5 e Megalina sono entrambi espressi in tutte le biopsie microdissezionate sia a livello glomerulare che a livello tubulo interstiziale e i loro livelli di espressione erano simili nei due compartimenti in entrambe le patologie; 2) i livelli di espressione sono risultati maggiori nelle nefropatie proteinuriche rispetto ai controlli sia a livello tubulo interstiziale che a livello glomerulare. Per escludere un’eventuale contaminazione del compartimento glomerulare da parte del tubulo prossimale abbiamo chiesto la collaborazione del Dott. J.J.Baelde del Leiden University Medical Center che ha effettuato l’analisi di espressione di CLCN5 e Megalina su glomeruli microdissezionati mediante laser ottenuti da biopsie di pazienti NIDDM (n°28) e su glomeruli sani di controllo (n°14), confermando i nostri precedenti risultati. In queste biopsie microdissezionate è inoltre emersa una correlazione diretta tra CLCN5 e Megalina (r=0,5, p=0,0039); 3) la presenza di CLCN5 è stata evidenziata e quantificata in tutte le cellule analizzate, mentre per Megalina sono stati rilevati livelli molto bassi di espressione; 4) le cellule HK-2 stimolate con albumina hanno mostrato gli stessi livelli di espressione di CLCN5 delle cellule non trattate; 5) le cellule HMC stimolate con glucosio hanno mostrato gli stessi livelli di espressione di CLCN5 delle cellule non trattate. Il nostro studio evidenzia per la prima volta la presenza di CLCN5 e Megalina a livello del compartimento glomerulare nell’uomo ed inoltre che tutti i tipi cellulari costituenti il glomerulo esprimono in vitro CLCN5. La sovrespressione di CLCN5 e Megalina nelle biopsie dei pazienti diabetici e con nefropatia da IgA suggerisce un loro ruolo nella fisiopatologia della proteinuria e la loro correlazione diretta, emersa tra l’espressione genica delle due proteine nel glomerulo, fa ipotizzare un loro meccanismo d’azione coordinato simile a quello presente nel tubulo prossimale. Gli esperimenti in vitro sulle HMC escludono un ruolo del glucosio nella modulazione dell’espressione di CLCN5 nella patologia diabetica. Un ruolo in questo processo potrebbe essere ipotizzato per i podociti in quanto i risultati da noi ottenuti bene si affiancano ai dati in letteratura che mostrano che i podociti, nei pazienti proteinurici, sono in grado di endocitare proteine.

Megalin is an endocytotic receptor belonging to the low-density lipoprotein family. It has often been viewed only as merely a scavenger receptor of absorptive and secretory epithelia. Recent work has revealed that the megalin intracellular domain contains several motifs potentially binding proteins involved in signal transduction.

To find potential intracellular proteins binding to megalin, a yeast two-hybrid screening was initiated with the intracellular tail of megalin as the bait. A partial clone encoding the scaffolding protein postsynaptic protein 95 (PSD-95) was found to bind to megalin with its second PDZ-domain. Co-localization experiments in HEK-293 cells and kidney, placenta and parathyroid tissue confirmed this interaction. The PSD-95 related proteins PSD-93 and SAP102 were also confirmed to bind megalin with their PDZ2-domains, but the corresponding domain from SAP97 did not bind. Mutation analysis revealed that an amino acid residue change Ala to Thr was the cause of this.

Megalin has within the central nervous system (CNS) been shown to be expressed only in the ependymal cells and choroid plexus. Nothing has been known about megalin expression in the spinal cord. To study spatio-temporal expression of megalin in the spinal cord, extensive staining of prenatal and postnatal mouse spinal cord was undertaken. Megalin expression was found in the dorsal part of the embryonic spinal cord. Most of these cells also expressed vimentin, suggesting that megalin has a role in the normal development of astrocytes. In the postnatal mouse, megalin seems to be expressed in oligodendrocytes only in the spinal cord white matter, and co-incident with myelination. This suggests that megalin is involved in the formation and maintenance of myelin along long spinal pathways. Megalin staining was clearly seen in the nucleus of these cells, indicating that megalin works in a notch-like signalling pathway.

Uptake of retinol to the retina pigment epithelium (RPE) has long been thought to be a diffusion process. Staining for megalin in RPE revealed strong expression, and uptake experiments with 3H-retinol bound to retinol-binding protein and blocking with the LDL-receptor family specific antagonist receptor-associated protein (RAP) showed that megalin is a receptor for uptake of retinol to the RPE.

Background: Intra-amniotic infections (IAIs) are common complications of labor and delivery. If inadequately treated, these infections can lead to significant morbidity and mortality in the mother and the fetus. Intrapartum aminoglycoside (AG) administration is recommended for the management of IAIs. AGs are known to cross the placenta and achieve bactericidal concentrations in fetal serum. However, the highest and most persistent fetal levels are achieved in renal tissue. So, the fetus may be vulnerable to the nephrotoxic effects of AGs. Megalin, a 600 kDaendocytic receptor, is responsible for the uptake of AGs into renal proximal tubular epithelial cells. This receptor is also expressed in human term placenta and it is reasonable to speculate that it is similarly involved in the placental transport of AGs. However, the mechanisms responsible for placental AG uptake and transport have not yet been characterized. Objective: To evaluate the role of megalin in the transport of AGs across human placenta. Specific aims: (1) To assess and compare megalin expression in term and preterm placental villous tissue, and (2) assess the functional activity of megalin in in vitro placental models. Methods: (1) Following IRB approval, placental tissue samples were collected from pregnant women undergoing term or preterm deliveries. Placental villous tissueswere used to quantify megalin expression by western blotting and q-PCR (2) The human choriocarcinoma cell line (BeWo cells) were grown on Transwell plates, and then megalin expression and function were assessed. Results: Megalin protein and mRNA expression were confirmed in samples of human placental villous tissues. Megalin mRNA expression declined steeply with gestational age till week 31 of gestation then it plateaued thereafter. Also, the expression in the early preterm (n=2) was six fold higher than that of both late preterm (n=3) and term placenta (n=10) (p<0.05). The uptake of 3H-gentamicin by the BeWo cells was time-dependent, saturable (Vmax=42.9 ± 4.9 nmol/mg protein/min; Km=2.93±0.68mM) and partially inhibited by megalin inhibitors. Conclusion: Megalin is expressed in human placental villous tissues as well as the BeWo cells. When grown on Transwell® plates, the BeWo cells appear to be the most appropriate model to study the in vitro transport of AGs across the apical membrane. Time, temperature and concentration dependence of gentamicin uptake in the BeWo cells indicate protein-mediated transport. The inhibition data are consistent with megalin-mediated endocytosis of AGs.

Der Endozytoserezeptor Megalin gehört zu einer Gruppe von strukturell und funktionell verwandter Rezeptoren, der LDL R Gen Familie. Es sind zwei Arten von Lipidtransportpartikel beschrieben worden, die durch Megalin in Zellen aufgenommen werden. Zum einen werden Lipoproteine über ihre Apoproteine von Megalin erkannt und endozytiert. Zum anderen nimmt Megalin die hydrophoben Vitamine A und D über ihre Carrierproteine in ihre Zielzellen auf. Es handelt sich um Vitamin D bindendes Protein (DBP) und Retinol bindendes Protein (RBP). Zweck dieser Arbeit war es zu untersuchen, ob die Endozytose von Steroidcarriern durch Megalin ein genereller Mechanismus ist oder ob DBP und RBP Ausnahmen darstellen. Hierzu wurden exemplarisch drei Carrierproteine (24p3, Apo D und CCSP) für Steroide ausgesucht, die in Megalin-exprimierende Gewebe aufgenommen werden. Der (rekombinante) Retinolcarrier 23p3 zeigte bei surface plasmon resonance Analysen keine direkte Bindung an Megalin. Der Progesteron-Carrier Apo D hingegen bindet Megalin, ferner konnte in Zellkulturversuchen Endozytose und lysosomale Degradation von Apo D in Megalin-exprimierende Zellen nachgewiesen werden. Auch der Progesteron-Carrier CCSP wird durch Megalin in Zellen aufgenommen, allerdings ist zur Endozytose von CCSP ein Co-Rezeptor notwendig. Mit dieser Arbeit ist die erste in vivo-Beschreibung eines dualen Rezeptorsystems aus Megalin und einem peripheren Membranprotein namens Cubilin gelungen, welches u.a. im proximalen Tubulus der Niere existiert. Abschließend wurde exemplarisch für ein Steroidhormon-abhängiges Gewebe der murine Uterus hinsichtlich seiner Megalin-Expression untersucht. Es konnte ein bereits bekannter Ligand Megalins, das Glykoprotein Laktoferrin, aus der uterinen, luminalen Flüssigkeit aufgereinigt werden. Ferner konnte gezeigt werden, dass die Expression von Laktoferrin im Uterus strenger hormoneller Kontrolle unterliegt.
The endozytic receptor Megalin belongs to a group of structurally and functionally related receptors called LDL R gene family. Two different types of lipid particles are taken up by Megalin into target cells. The first type, lipoproteins are recognized and internalized by Megalin via their apoproteins. In addition, Megalin mediates the endocytosis of the lipophilic vitamins A and D into target cells by means of their carrier proteins. These proteins are the vitamin D binding protein (DBP) and retinal binding protein (RBP). The aim of the investigations was to determine, if the endocytosis of steroid hormone carriers by Megalin is a common occurrence or restricted only to DBP and RBP. Therefore, three carrier proteins for steroids (24p3, Apo D and CCSP) were chosen as an example. All of them are known to be taken up in Megalin expressing tissues. In surface plasmon resonance analysis recombinant 24p3, a carrier of retinol, showed no affinity to Megalin. Whereas the progesterone carrier Apo D bound to Megalin. Furthermore, it was endozytosed and degraded in lysosomes by Megalin expressing cells. The cellular uptake of the progesterone carrier CCSP is mediated by Megalin as well, however a co-receptor is needed. This work demonstrates for the first time the existence of a dual receptor pathway consisting of Megalin and a peripheral membrane protein named Cubilin in vivo. This systems is functional in addition to other tissues in the proximal tubule of the kidney. Finally, the Megalin expression in the murine uterus as an example of a steroid dependent tissue was investigated. Lactoferrin a known Megalin ligand was purified from the luminal uterine fluid. Furthermore, Lactoferrin expression in the uterus was shown to be under tight hormonal control.

LRP2 wird waehrend der fruehen Embryonalentwicklung hauptsaechlich im Dottersack und im Neuroepithel exprimiert. Der funktionelle Verlust dieses 600 kDa grossen Proteins in Maeusen fuehrt zu Fehlbildungen bei der Vorderhirnentwicklung, die als Holoprosenzephalie bezeichnet werden. LRP2 kommt daher eine wichtige Funktion waehrend der Vorderhirnentwicklung zu. Fuer diese Arbeit wurden Maeuse verwendet, bei denen lrp2 im gesamten Tier oder nur in bestimmten Geweben inaktiviert war. Die Expression von LRP2 im Neuroepithel, nicht aber im Dottersack ist wichtig fuer die korrekte Vorderhirnentwicklung. Das Fehlen des Proteins fuehrte am Tag 9.5 der Embryonalentwicklung zu einer UEberaktivierung des Bmp4 Signalweges im rostralen Telencephalon. Am Tag 10.5 war ein Verlust der shh-Expression in der anterioren entopeduncularen Zone (AEP) zu sehen. Das Fehlen von Shh in der AEP fuehrte zum Verlust von ventralen Oligodendrozyten und Interneuronen. AEhnliche Defekte wurden ebenfalls in Maeusen beschrieben, bei denen der Bmp4 Signalweg verstaerkt ist. Bmp4 bindet in vitro an LRP2. Diese Ergebnisse deuten darauf hin, dass LRP2 an der Entwicklung des ventralen Telencephalons beteiligt ist - moeglicherweise indem es die verfuegbare Menge an Bmp4 durch Endozytose reguliert. Die Signalwege, in die LRP2 eingebunden ist, sollten im Zebrafisch weiter untersucht werden. Tiere, denen funktionelles LRP2 fehlte, wurden durch die Injektion von Morpholinos generiert. Injizierte Tiere zeigten Stoerungen bei Resorptionsvorgaengen im Pronephros, was darauf hin deutet, dass die Funktion von LRP2 im Zebrafisch Pronephros und in der Niere der Saeugetiere konserviert ist. Die Expressionsmuster von Markergenen fuer die Vorderhirnentwicklung waren allerdings normal.Vermutlich war die Stoerung der lrp2-mRNA Prozessierung nicht ausreichend, um hier einen Effekt hervorzurufen, da bereits im 1-Zell Stadium genuegend komplett prozessierte maternale lrp2-mRNA fuer die Translation von LRP2 vorhanden war.
LRP2 is mainly expressed in the yolk sac and in the neuroepithelium of the early embryo. Deficiency for this 600 kDa protein in mice results in holoprosencephaly, indicating a role for LRP2 in forebrain development. Mice with a complete or a conditional loss of lrp2 function were used to elucidate the consequences of the lack of LRP2 expression. The presence of LRP2 in the neuroepithelium but not in the yolk sac is crucial for early forebrain development. Lack of the receptor resulted in an increase of Bmp4 signaling in the rostral telencephalon at E9.5. As a consequence, shh expression at E10.5 was lost completely in a ventral region of the telencephalon termed anterior entopeduncular area (AEP). The absence of Shh activity in this area led to the loss of ventrally induced oligodendroglial and interneuronal cell populations in lrp2-deficient mice. Similar dorsalizing effects have also been observed in mice with increased Bmp4 signaling. Taking into account that Bmp4 was found to bind to LRP2 in vitro and in vivo, these results suggest a role for LRP2 in patterning the rostral ventral neural tube, possibly by acting as a clearance receptor for Bmp4. The underlying molecular mechanisms were then further analyzed in the zebrafish. LRP2-deficient animals were generated by injecting Morpholinos that interfered with the splicing of the lrp2-pre-mRNA leading to a deletion of the transmembrane-exon. Injected animals suffered from impaired renal clearance processes, demonstrating the functional conservation of LRP2 in the larval zebrafish pronephros. Brain structures were not affected in these animals and the expression patterns of marker genes for early forebrain development that were changed in the mouse were not changed. Apparently, Morpholino mediated interfering with the splicing of the lrp2-pre-mRNA did not affect the early forebrain formation because properly processed lrp2-mRNA was supplied maternally and sufficient for proper brain formation.

The construction of archaeological narrative is influenced by a number of factors. Some come from within disciplinary boundaries, whilst others are traced from the wider influences of social, cultural or academic contexts. This thesis examines three areas identified as Neolithic ‘landscapes’, all of which have been the subject of archaeological investigation since the 19th century. The history of research of these areas allows an evaluation of how these disparate influences interact. In this way, the three landscapes act as an arena in which to explore aspects of the archaeological approach itself. This leads to a critical examination of the interpretative tools available to the archaeologist. How concepts such as ‘landscape’ are formed and affect discourse is explored. Wider themes of demarcation, typology and the underlying assumptions of research are investigated in relation to the interpretation of the Neolithic and Early Bronze Age of North Western Europe. The large span of time that these periods encompass allows exploration of change from the short to very long term, although this is not always utilised within archaeological accounts. The treatment of time is therefore considered in conjunction with explanations of change in prehistory. A powerful approach to time is suggested by combining aspects of the work of Pierre Bourdieu and Fernand Braudel and the potential for this is evaluated against the archaeological record of the three areas. How the assumptions of the archaeological approach are acted out within the historiography of each area highlights a number of recurring metaphors that are used to interpret the material record. These promote a portrayal of Neolithic life that combines with the range of influences from the history of archaeology itself to promote an idea of the prehistoric mentalité. A very durable and underlying type that constantly resurfaces in these accounts is the idea of the ‘sacral landscape’, which is the central topic of this thesis.

Cubilin is a peripheral membrane protein that co-operates with the endocytic receptor megalin to mediate the endocytosis of ligands in various polarized epithelia. Megalin is expressed in the male reproductive tract where is has been implicated in the process of sperm membrane remodeling. A potential role for cubilin in the male reproductive tract has not been explored. Using RT-PCR, we found that cubilin and megalin mRNAs are expressed in the efferent ducts, corpus and cauda epididymides, and proximal and distal vas deferens. Immunohistological analysis revealed that cubilin was expressed in non-ciliated cells of the efferent ducts, principal cells of the corpus and cauda epididymides and vas deferens. Electron microscopic immunogold labeling showed cubilin in endocytic pits, endocytic vesicles and endosomes of these cells. The expression profile of cubilin in the male reproductive tract was coincident with that of megalin except in principal cells of the caput epididymidis. Double immunogold labeling showed that cubilin and megalin co-localized with one another within the endocytic apparatus and recycling vesicles of efferent duct cells. Neither protein was found in lysosomes. Injection of RAP, an antagonist of megalin interaction with cubilin, reduced the level of intracellular cubilin in cells of the efferent ducts and vas deferens. In conclusion, cubilin and megalin are co-expressed in cells of the epididymis and vas deferens and the endocytosis of cubilin in these tissues is dependent on megalin. Together, these findings highlight the potential for a joint endocytic role for cubilin and megalin in the male reproductive tract.

This report documents the development of a concept and prototype for a mobile application with the purpose of making the megalith graves surrounding the town of Falköping more engaging and interactive for visitors. Because of the limitations of working with heritage sites that cannot be altered to support a gaming experience, the usage of escape room game mechanics is explored. Because escape room games depend on the user's observation and exploration of their environment, they serve as a usable inspiration for applications with the purpose of making visitors experience a location in a similar way. The prototype developed for the project contains puzzles designed for four different megalith grave sites which were then tested with groups of children and parents playing together.

Proteinuria is clearly associated with the progression of chronic kidney disease (CKD) but the mechanisms underlying this relationship remain unclear. Recent evidence suggests that altered proximal tubular (PT) handling of filtered proteins may significantly modulate urine protein excretion and progressive renal disease. Megalin and neonatal Fc receptor (FcRn) are endocytic receptors responsible for the PT reabsorption of glomerular ultrafiltered proteins by receptor-mediated endocytosis (RME) and transcytosis, and are expressed on the luminal surface of the PT. This study aimed to investigate the expression and turnover of megalin and FcRn in proteinuria, and the possible mechanisms underlying down-regulation of expression of these receptors. In protein overload proteinuria (POP), the PT expression of megalin and FcRn were both downregulated compared to control animals. In addition, significant amount of both receptor were detected in the urine of proteinuric animals. This reduction in megalin and FcRn protein expression was accompanied by a significant increase in the urinary excretion of plasma proteins including the most abundant albumin. Increased expression and activity of both matrix metalloproteinase (MMP) and gamma secretase (γ-secretase) was evident in proteinuric animals and was paralleled with the urinary excretion of both megalin and FcRn. Megalin was previously identified to undergo regulated intramembrane proteolysis (RIP). RIP includes the MMP-mediated cleavage of the extracellular domain of megalin which in turn excreted into the urine, and the subsequent cleavage of the intracellular domain by γ-secretase. A synthetic inhibitor of MMPs, batimastat (BB-94) significantly reduced urinary excretion and also preserved the PT expression of megalin. In addition, inhibition of MMPs appeared to be antiproteinuric by markedly reducing urinary excretion of albumin and plasma proteins. The urinary excretion of FcRn was also reduced in BB-94 treated animals but the mechanism of this reduction was not clear. Further, BB-94 treatment preserved PT expression of FcRn in proteinuric animals. The POP model was developed to investigate the degree of tubulointerstitial damage in experimental animals. Injury was detected in the interstitium of proteinuric animals with significant increases in inflammatory markers including tumour necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and the transforming growth factor-beta (TGF-β) as well as interstitial collagen deposition and marked interstitial infiltration of macrophages (F4/80). Blockade of MMP in proteinuric animals significantly reduced the expression of these markers in the kidney and abrogated the progression of interstitial inflammation and fibrosis. In summary this is the first study to correlate the reduced expression of megalin and FcRn, and the urinary excretion of these receptors with the increased cellular activity of MMP and the possible occurrence of RIP in vivo. Inhibition of MMP could be of great importance in treating proteinuria and the progression of tubulointerstitial inflammation and fibrosis, the most characteristic features of CKD.

This essay studies the funnel-beaker ceramic in connection with the megalith graves in South Scandinavia. In my work I have described the nature of ceramics and looked on the crockery types and ornamentation and then discussed the use of the material in a ritual perspective. In the first part I have focused on the critical aspects you have to think about when you do a study like this one. In the second part of this essay I have done a case study, with the ceramic material, in three passages graves in the area of Falbygden, Western Sweden and studied the ceramic material and the nature of it in South Scandinavia as well. Then in the third I discussed the potential use, there is two, of the ceramic material in general of South Scandinavia and looked at it in a ritual perspective.

ClC-5 with megalin (LRP2), cubilin, Disabled 2 (Dab2), and Amnionless (AMN), is part of the molecular complex involved at proximal tubular level in the endocytic re-uptake of low-molecular-weight proteins and albumin. ClC-5, megalin and cubilin expression in podocytes of human renal biopsies was already reported. Moreover, it was demonstrated that podocytes are able to internalize albumin through an endocytic process. It is reasonable to assume that there should be an involvement of this system in protein uptake mediated by podocytes. A disruption of this system can lead to proteinuria which is one of the first manifestation of kidney disease in Systemic Lupus Erythematous (SLE). Aims of this study were to explore the presence of the tubular endocytic machinery components also in human podocytes in vitro and to evaluate whether albumin modulates this system. Moreover, we wanted to verify and quantify the expression of ClC-5, megalin and cubilin in both glomerular and tubular compartments in renal biopsies of patients presenting Lupus nephritis and observing the presence of a relationship with clinical data. To verify the presence of an uptake mechanism in human podocytes in vitro, we performed time lapse experiments with a low dose of FITC albumin (10 µg/ml). We observed albumin internalization starting from 2 to 15 hours. To evaluate the uptake kinetic, we stimulated podocytes at different time (30 min and 2 hours) and doses (10 µg/ml, 100 µg/ml and 1 mg/ml) at 37°C and 4°C. We observed a significant dose dependent increase in fluorescence vs controls after 2 hours’ stimulation with a typical receptor-mediated kinetic since it is inhibited at 4 °C. Moreover, we disclosed the presence of ClC-5, Dab2 and AMN beyond megalin and cubilin, in this in vitro system using immunohistochemistry (IHC) and immunofluorescence (IF) techniques, highlighting a co-localization between albumin and both receptors. To analyze whether the proteinuric environment modulates CLCN5, LRP2, CUBN, DAB2 and AMN expression, we stimulated human podocytes with increasing concentrations of BSA (range 10 µg/ml - 30 mg/ml) and we evaluated the mRNA expression at different time points (2, 4, 8, 24, 48 and 72 hours). Using Real Time PCR we observed a significant time and dose dependent increase in CLCN5, CUBN and AMN expression and an up-regulation of DAB2 only at 24 hours. We collected 23 SLE renal biopsies, 6 control biopsies and 1 case of Minimal Change Disease. As clinical parameters we considered proteinuria and pharmacological therapy. IHC and IF were used to analyze ClC-5, megalin and cubilin protein expression in serial sections. Morphometric quantification revealed a direct correlation between tubular and glomerular expression of all molecules in SLE patients, highlighting a relationship between glomerular and tubular compartments independently from proteinuria levels. Furthermore, preliminary data on patients without ACEi/ARB and immunosuppressive drugs disclosed a positive trend among these molecules at glomerular level. Interestingly, we revealed megalin and cubilin expression in hypertrophic PECs of some SLE patients and characterization experiments identified a subpopulation with an intermediate phenotype between mature and progenitor cells. In conclusion, for the first time we demonstrated that human podocytes are naturally committed to perform albumin endocytosis via a receptor-mediated mechanism. Moreover, protein overload upregulates CUBN, AMN and CLCN5 in these cells. Functional studies regarding the role of cubilin in albumin uptake underlined its participation in this mechanism even if this is not the only pathway involved. Further studies will be necessary to analyze which may be the partner(s) of the CUBAM complex in this mechanism. For the first time we demonstrated the presence of ClC-5, megalin and cubilin in glomeruli of patients with SLE and MCD in addition to controls confirming in vitro data. Furthermore, in SLE biopsies we highlighted a strong correlation between the two renal compartments in the expression of the protein uptake system, supporting the idea of a partnership between tubular and glomerular cells in albumin uptake via the same mechanism of internalization. In addition, preliminary data on patients without ACEi/ARB and immunosuppressive drugs lead us to suppose that the pharmacological therapy could affect the expression of this system, in particular at glomerular level. The differences observed from in vivo and in vitro data, especially on megalin expression, underline the involvement of other glomerular cell types in addition to podocytes in protein uptake. Finally, megalin and cubilin expression in PECs of SLE patients is a real interesting but complex data, since characterization experiments identified a subpopulation with an intermediate phenotype between mature and progenitor cells. Further studies will be performed to better characterize the role of these double-positive cells and their correlation with clinical data and/or disease progression.
ClC-5, megalina (LRP2), cubilina, Disabled 2 (Dab2) ed Amnionless (AMN) fanno parte del complesso molecolare coinvolto a livello del tubulo prossimale nel recupero delle proteine a basso peso molecolare e dell’albumina mediante endocitosi. È già stata riportata la presenza di ClC-5, megalina e cubilina a livello dei podociti in biopsie renali umane. Inoltre è stato dimostrato che i podociti sono in grado di internalizzare l’albumina attraverso un meccanismo di endocitosi. Il mancato funzionamento di questo sistema può portare a proteinuria, che è una delle prime manifestazioni del coinvolgimento renale nel Lupus Eritematoso Sistemico (LES). È quindi ragionevole supporre che vi possa essere un coinvolgimento di questo sistema nel meccanismo di uptake delle proteine da parte dei podociti. Gli scopi di questo studio sono stati di esplorare la presenza dei componenti del sistema tubulare di endocitosi delle proteine in podociti umani in coltura e di valutare se e come l’albumina ne modulasse l’espressione. Inoltre, si è voluto indagare l’espressione di ClC-5, megalina e cubilina sia a livello glomerulare che tubulare in biopsie renali di pazienti con nefrite lupica, valutando una possibile relazione con i dati clinici. Abbiamo verificato la presenza di un meccanismo di uptake in podociti umani in coltura attraverso esperimenti di time-lapse con basse dosi di FITC-BSA (10 µg/ml) ed abbiamo osservato l’inizio del processo di internalizzazione in un periodo di tempo variabile dalle 2 alle 15 ore. Per caratterizzare il tipo di cinetica di uptake della FITC-BSA, i podociti sono stati stimolati a differenti tempi (30 min e 2 ore) e dosi (10 µg/ml, 100 µg/ml and 1 mg/ml) mantenendo la coltura a 37°C o 4°C. Si è osservato un aumento significativo della fluorescenza dose-dipendente rispetto al controllo dopo 2 ore dalla stimolazione con una tipica cinetica di internalizzazione recettore-mediata poiché veniva inibita a 4°C. Abbiamo osservato la presenza di ClC-5, Dab2 e AMN oltre a quella di megalina e cubilina in podociti umani in coltura in condizioni basali mediante tecniche di immunoistochimica (IHC) ed immunofluorescenza (IF) ed abbiamo dimostrato la co-localizzazione dei due recettori con l’albumina fluorescente. Per valutare se l’ambiente proteinurico fosse in grado di modulare l’espressione di CLCN5, LRP2, CUBN, DAB2 ed AMN, i podociti umani sono stati stimolati con concentrazioni crescenti di BSA (range 10 µg/ml - 30 mg/ml) e l’espressione dell’RNA messaggero è stata valutata a tempi diversi (2, 4, 8, 24, 48 and 72 hours). Mediante analisi in Real Time PCR, abbiamo osservato un aumento significativo tempo e dose-dipendente di CLCN5, CUBN ed AMN ed un aumento di DAB2 solamente alle 24 ore. Abbiamo raccolto 23 biopsie renali di pazienti con LES, 6 biopsie di controllo ed un caso di Minimal Change Disease. Come parametri clinici abbiamo considerato la proteinuria e la terapia farmacologica. Mediante IHC ed IF abbiamo analizzato l’espressione proteica di ClC-5, megalina e cubilina in sezioni seriali. La quantificazione eseguita mediante analisi morfometrica ha rivelato una correlazione diretta dell’espressione di tutte le molecole in analisi tra il compartimento tubulare e glomerulare, evidenziando una stretta relazione tra i due compartimenti indipendentemente dai livelli di proteinuria. Inoltre, dati preliminari su pazienti privi di terapia farmacologica (ACEi/ARB o immunosoppressivi) hanno mostrato un trend positivo tra l’espressione di queste molecole a livello glomerulare. Curiosamente, abbiamo evidenziato l’espressione di megalina e cubilina in cellule parietali della capsula (PECs) con morfologia ipertrofica in alcuni pazienti LES che, mediante esperimenti di caratterizzazione, abbiamo identificato come una nuova sottopopolazione con un fenotipo intermedio tra cellule mature e progenitrici. Concludendo, per la prima volta abbiamo dimostrato che i podociti umani sono naturalmente predisposti ad effettuare l’endocitosi dell’albumina attraverso un meccanismo recettore-mediato. Inoltre, l’overload proteico è in grado di aumentare l’espressione di CLCN5, CUBN ed AMN in queste cellule. Studi funzionali per dimostrare il ruolo di cubilina nel processo di uptake dell’albumina hanno sottolineato la sua partecipazione in questo meccanismo anche se, verosimilmente, non è l’unico pathway coinvolto. Ulteriori studi saranno necessari per analizzare quali altre molecole possano essere chiamate in causa in questo meccanismo. Per la prima volta abbiamo dimostrato la presenza di ClC-5, megalina e cubilina in glomeruli di pazienti con LES, MCD e controlli, confermando i dati in vitro. Inoltre, nelle biopsie dei pazienti LES abbiamo evidenziato una stretta relazione tra i due compartimenti renali nell’espressione dei componenti di questo sistema, supportando l’idea di una partnership tra cellule tubulari e glomerulari nell’uptake dell’albumina attraverso lo stesso meccanismo di internalizzazione. In aggiunta, dati preliminari ottenuti da pazienti privi di terapia con ACEi/ARB o immunosoppressivi ci ha fatto supporre che il trattamento farmacologico possa influire sull’espressione di questo sistema a livello glomerulare. Le differenze osservate tra lo studio in vivo e quello in vitro, in particolare riguardo l’espressione di megalina, suggeriscono il coinvolgimento di altre cellule del glomerulo oltre ai podociti. Infine, l’espressione di megalina e cubilina nelle PECs dei pazienti LES è un dato molto interessante ma complesso, poiché gli esperimenti di caratterizzazione hanno identificato una sottopopolazione con un fenotipo intermedio tra cellule mature e progenitrici. Ulteriori studi dovranno essere condotti per meglio caratterizzare il ruolo di queste cellule con doppia positività e la loro correlazione con i dati clinici o di progressione della malattia.

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This thesis provides an algorithm based on marginal analysis to help CLF Supply Officers load ships with Provisions (food), High Usage Load List (HULL), and ship store stock. The algorithm is incorporated into an EXCEL spreadsheet and produces a prioritized list of items in their optimum stocking sequence. The algorithm is compared to both the Atlantic and Pacific Fleet current loading methods using a food item example and shows a 60-70% increase in expected supply effectiveness. Furthermore, the algorithm is generic and can be applied to and inventory problem where a demand distribution is known and space is the limiting factor in determining stocking levels.

La mégaline est un récepteur endocytique multiligand essentiellement connu comme récepteur clé pour la réabsorption protéique rénale. Au cours du développement embryonnaire murin la mégaline a été impliquée dans le développement du prosencéphale et de l’œil mais la mortalité périnatale des souris Meg-/- empêche sone étude à des stades ultérieurs. Pour analyser le rôle de la mégaline dans le système visuel nous avons généré dans le laboratoire des souris conditionnelles FoxG1-Cre MegL/L.L’analyse phénotypique des souris mutantes montre que l’absence de la mégaline dans le neuroépithélium antérieur conduit à une augmentation anormale et progressive de la taille de l’œil accompagnée par un amincissement de la rétine et la diminution, entre autres, des cellules ganglionnaires. Ces observations immunomorphologiques suggèrent que les souris mutantes présentent des signes de myopie forte et sont confirmées par l’analyse ophtalmologique. Les analyses biochimiques des souris mutantes montrent que l’absence de mégaline entraine des modification dans la structure et la fonction du corps ciliaire, à l’origine de cet élongation excessive du globe oculaire. Les résultats obtenus montent que les souris Foxg1-Cre MegL/L sont le premier modèle génétique murin qui reproduit les caractéristiques de la myopie forte. L’élucidation des mécanismes physiopathologiques sous-jacents au développement de la myopie est un préalable indispensable à l’élaboration d’en éventuel traitement préventif, espoir pour les 0,5% à 2% de la population touchés par la cécité liée aux complications de la myopie forte
Megalin is a large multiligand endocytic receptor essentially known as the key receptor for protein reabsorption in the renal proximal tubule. Murin megalin appears to be essential for early forebrain and eye development but the perinatal lethality of Meg-/- mice precludes further analysis. To study the implication of megalin in the formation of the eye we generated (in the laboratory ) FoxG1-Cre MegL/L mice lacking megalin specifically in the anterior neuroepithelium.The phenotypic analysis of these mice show that they suffer from an excessive and progressive eye growth associated with retinal thinning and a strong decrease of some retinal cell types including the ganglion cells. The immunomorphological studies suggest that the mutant mice are highly myopic which is confirmed by the ophtalmological analysis data. The biochemical analysis of these mutant mice shows that the absence of megalin causes structural and functional modifications in the ciliary epithelium, leading to the excessive elongation of the eyeball. These results show that the FoxG1-Cre MegL/L mice are the first murine genetic model recapitulating the characteristics of high myopia. The elucidation of underlying physiopathological mechanisms of high myopia development is essential for the elaboration of a preventive treatment, hope for 0,5 to 2% of the population suffering from blindness caused by the complications of high myopia

Le mégalithisme est l'expression architecturale monumentale d'un ensemble de sociétés qui ont édifiées des structures funéraires et des pierres dressées. La genèse du mégalithisme, phénomène humain de la fin de la préhistoire, puise ses origines dans de multiples conjugaisons des différents courants de la néolithisation aux traditions variées, aux influences géographiques parfois lointaines, aux époques différentes. La combinaison de nombreux éléments sociaux de ces sociétés agro-pastorales a généré des générations d’architectes bâtisseurs et contribué à mettre en œuvre une diversité architecturale La volonté de mise œuvre des maisons des morts au même niveau que les maisons des vivants semble répondre à des critères sociaux et culturels. Une perception visuelle actualisée des mégalithismes, adaptée aux récentes thématiques développées amène un fil conducteur original d’étude de plusieurs mondes qui se sont combinés. L’état des connaissances depuis le milieu du XXème siècle a permis une appréhension différente des mégalithismes : - dans les années 1980, une connaissance plus complète des masses tumulaires et leurs liens étroits avec les chambres sépulcrales (Joussaume, 1997, 1999, 2003 ; Joussaume et al. 2006). - dans les années 2000 une prise en compte de l’histoire des monuments (Joussaume et al. 2006 ; Laporte, 2010 ; Laporte et al. 2004, 2011). - de nos jours, une appréhension différente des articulations entre les pierres dressées, les tumulus et les chambres sépulcrales (Laporte, 2015 ; Laporte et al. 2011). Le processus de cette monumentalisation architecturale a souvent fait l’objet d’études distinctes, les espaces sépulcraux et les pierres dressées servant de bases à deux axes de recherches séparées. Seule l’étude des stèles en remploi avait fait l’objet d’une attention particulière (L’Helgouac’h, 1983 ; Cassen, 2009b) et d’un rapprochement des deux dispositifs. Ce n’est que récemment que la complémentarité entre les espaces sépulcraux et les pierres dressées a réellement été proposée (Laporte, 2015b). Disposant d’un important corpus actualisé des mégalithes du département du Morbihan, il a donc été proposé de développer dans cette thèse cette notion de complémentarité entre les différents dispositifs qui constituent les mégalithismes. Les hypothèses formulées sont de démontrer que les processus de monumentalisation sont issus d’un croisement architectural entre les pierres dressées, les espaces sépulcraux et les masses tumulaires. Nous tenterons également de montrer les liens étroits qui semblent se dessiner entre trois mondes très différents mais intimement liés (le monde des vivants, le monde des morts et le monde naturel)
Megalithism is the monumental architectural expression of a group of societies that have built funerary structures and erected stones. The genesis of megalithism, the human phenomenon of the end of prehistory, draws its origins from multiple conjugations of the different currents of neolithization with varied traditions, geographical influences sometimes distant, at different times. The combination of many social elements of these agro-pastoral societies has generated generations of architects and contributed to the implementation of architectural diversity The desire to put the houses of the dead to the same level as the houses of the living seems to meet social and cultural criteria. An updated visual perception of megalithisms, adapted to the recent themes developed, brings an original thread of study of several worlds that have combined. The state of knowledge since the middle of the 20th century allowed a different apprehension of megalithisms: - in the 1980s, a more complete knowledge of the tumular masses and their close links with the sepulchral chambers (Joussaume, 1997, 1999, 2003, Joussaume et al., 2006). - in the 2000s, taking into account the history of monuments (Joussaume et al., 2006, Laporte, 2010, Laporte et al., 2004, 2011). - today, a different apprehension of the articulations between the erected stones, the tumuli and the sepulchral chambers (Laporte, 2015, Laporte et al., 2011). The process of this architectural monumentalization has often been the subject of separate studies, sepulchral spaces and erected stones serving as bases for two separate lines of research. Only the study of stelae in reuse had been the object of particular attention (L'Helgouac'h, 1983, Cassen, 2009b) and a comparison of the two devices. It is only recently that the complementarity between sepulchral spaces and erected stones has actually been proposed (Laporte, 2015b). It has therefore been proposed to develop in this thesis the notion of complementarity between the different mechanisms that constitute megalithisms. The hypotheses formulated are to demonstrate that the processes of monumentalisation are the result of an architectural crossing between the erected stones, the sepulchral spaces and the tumular masses. We will also try to show the close ties that seem to be taking shape between three very different but intimately linked worlds (the world of the living, the world of the dead and the natural world)

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FUJB
A ontogenia de Cryptodromiopsis antillensis (Stimpson, 1858) foi analisada através de dados de literatura (RICE & PROVENZANO, 1966) e observações da megalopa, juvenis e adultos. A megalopa de C. antillensis foi redescrita e Evius ruber Moreira, 1912 foi considerada como sinônimo de C. antillensis por tratar-se da fase megalopa desta espécie. A ontogenia da carapaça e dos apêndices cefálicos, torácicos e abdominais foi acompanhada separadamente. Carapaça, antênulas, antenas, mandíbulas, maxílulas, maxílas, primeiro maxilípede, segundo maxilípede, terceiro maxilípede e pereópodes sofrem mudanças significativas até a fase megalopa, quando adquirem uma forma similar a do adulto. Após a fase megalopa estes apêndices sofrem apenas acréscimo de cerdas. Já o esterno torácico, abdomen, telso, pleópodes e urópodes se modificam até a fase adulta, apresentando dimorfismo sexual. Pl 1 está ausente nas fases zoé e megalopa, surgindo apenas nos juvenis. Foi observada a presença de pleópodes rudimentares (Pl 3-Pl 5) nos espécimens macho de.. C. antillensis. A espécie C. antillensis se encaixa no padrão ontogenético da família Dromiidae. A única espécie que foge ao padrão dromiídeo é Cryptodromia tuberculata, que apresenta desenvolvimento abreviado. Dados ontogenéticos não suportam a monofilia dos Podotremata. A zoé 1 dos Dromiidae e a pré-zoé dos Dynomenidae são similares. A ontogenia do Homolodromiidae apresentou-se bastante diferente daquela dos Dromiidae. O caráter urópodes rudimentares (visíveis ventralmente) na fase megalopa, separa os Homolodromiidae de todos outros Podotremata. Apenas um caráter ontogenético sustenta os Dromiacea sensu GUINOT (1978): exopodito da antena desenvolvido na megalopa. O padrão ontogenético dos Archaeobrachyra sensu GUINOT (1978) também foi estudado. Dados ontogenéticos não fornecem suporte ao agrupamento dos Archaeobrachyura. Os seguintes caracteres ontogenéticos caracterizam os Homoloidea: 4-6 cerdas e estetos no protopodito da antênula na zoé 1; padrão de cerdas na base (2, 2, ,2, 3) e no endopodito (1+0, 1+0, 1+0, 2+0, 4+1) do Mxpl na zoé 1; endopodito do Mxpl divdido em dois artículos na megalopa; padrão de cerdas na base (1, 1, 1, 1) do Mxp2 na zoé 1. Apenas um caráter ontogenético é comum ao Raninidae e Cymonomidae: telso furcado.
The ontogeny of Cryptodromiopsis antillensis (Stimpson, 1858) it was analyzed through literature data (RICE & PROVENZANO, 1966) and observations of the megalopa, juveniles and adults. The megalopa of C. antillensis was redescribed and Evius ruber Moreira, 1912 was considered as synonym of C. antillensis by being of the phase megalopa of this species. The ontogeny of the carapace and the cephalic, thoracic and abdominal appendages it was accompanied separately. Carapace, antennule, antenna, mandible, maxillule, maxilla, first maxiliped, second maxiliped, third maxiliped and pereiopods suffer significant changes until the phase megalopa, when they acquire a similar form the one of the adult. After the phase megalopa these appendages just suffer increment of setae. Already the sternum thoracic, abdomen, telson, pleopods and uropods modify until the adult phase, presenting sexual dimorfism. Pl 1 is absent in the phases zoea and megalopa, just appearing in the juveniles ones. The presence of rudimentary pleopods was observed (Pl 3-Pl 5) in the male specimens of C. antillensis. Cryptodromiopsis. Antillensis, has the ontogenetic pattern of the family Dromiidae. The only species that outrange the Dromiidae pattern is Cryptodromia tuberculata, that presents abbreviated development. Ontogenetic data don't support the monophiletism of Podotremata. The zoea 1 of Dromiidae and the pre-zoea of Dynomenidae are similar. The ontogeny of Homolodromiidae came quite different from that of Dromiidae. The character rudimentary uropods (in a ventral view) in the megalopa phase, separates Homolodromiidae from all other Podotremata. Only one ontogenetic character justifies the Dromiacea sensu GUINOT ( 1978): antennal exopod developed in the megalopa. The ontogenetic pattern of the Archaeobrachyura sensu GUINOT ( 1978) was also studied. Ontogenetic data don't support the group of Archaeobrachyura. The following ontogenetic characters diagnoses Homoloidea: 4-6 setae and aesthetasc setae in the protopod of the antennule in the zoea 1; pattern of setae in the base (2, 2, ,2, 3) and in the endopod ( 1+0, 1+0, 1+0, 2+0, 4+1) of the Mxpl in the zoea 1; endopod of the Mxpl dividido in two articles in the megalopa; pattern of setae in the base ( 1, 1, 1, 1) of the Mxp2 in the zoea 1. Only one ontogenetic character is common to Raninidae and Cymonomidae: furcated telson.

Kidney disease is a major challenge for health care systems, and the prevalence is increasing. Proteinuria is a hallmark of progressive renal dysfunction and describes the pathological excess of plasma proteins in urine, mainly albumin. Multiple Myeloma is a cancer of plasma cells that leads to excessive presence of free light chain protein (FLC) in blood. Renal failure due to overproduction of FLC and the associated light chain proteinuria occurs as a result of decreased renal function or as a direct toxic effect on the proximal tubular cells (PTCs) by excessive protein. Proteins are normally reabsorbed by endocytosis via megalin receptor that binds proteins and mediates their uptake. Exceeding the proximal tubular epithelial cells (PTECs) reabsorption capacity might trigger inflammation detrimental to the kidney. In proteinuric nephropathy the cytoplasmic tail of megalin (MegCT) is phosphorylated after interaction between proteins and megalin on the PTECs, which activates signalling cascades that regulate the phosphorylation. An in vitro proteinuric model was established using HK2 cells (a proximal tubular epithelial cell line derived from normal human kidney) treated with high concentrations of essentially fatty acid free human serum albumin (FAF-HSA) or lambda light chain (λ-LC) isolated and purified from the urine of a myeloma patient, to induce cellular damage. The potential pathogenic role for FAF-HSA and λ-LC on HK2 cells was examined. Also, renal toxicity that comes from the intracellular signalling through phosphorylation of MegCT was addressed by utilising antibodies directed against specific phosphorylation site (PPPSP) of the intracellular portion of megalin in HK2 cells stimulated with different concentrations of FAF-HSA and λ-LC, so-called pre-stimulated HK2. In vitro analyses showed (i) a detrimental effect of FAF-HSA and λ-LC on viability of HK2, (ii) phosphorylation of the cytoplasmic tail of megalin in pre-stimulated HK2 cells. (iii) Production of inflammatory cytokines and H2O2 generation, activation of autophagy process and increase in several kidney biomarkers/ injury mediators, which are involved in different pathways in response to protein overload. All these reasons are likely to contribute to direct PTECs injury and kidney failure in patients. Potentially these mechanisms may be attractive for drug development to benefit patients with kidney failure and help to inhibit the progression of proteinuric nephropathy and as such may save lives.

Background The proliferation signal inhibitors (PSIs) sirolimus (SRL) and everolimus (ERL) are the potent immunosuppressive drugs using in organ transplantation and has been used successfully in renal transplant recipients (RTX) as well. PSIs are the key factors to overcome the allograft rejections after successful organ transplantation since the immune system starts to react against the graft. SRL and ERL prevents the action of immune system b inhibits the proliferation of T- and B-cells by inhibiting the intracellular signaling of interleukin-2. The presence of excess amount of serum proteins including albumin in the urine is considered as proteinuria, which reflects the loss of kidney function. The occurrence of proteinuria can be the result of abnormal glomerular filtration and/or impaired tubular endocytic function of renal proximal tubular epithelial cells (PTECs). Megalin and cubulin are two scavenger receptors present on epical surface of PTECs and involved in reabsorption of proteins after glomerular ultrafiltration process in the kidney. Proteinuria appears too high in renal transplanted patients during ongoing   treatment with PSIs.

Aim Our study aimed to investigate and correlate the expression level of megalin and cubilin and albumin uptake in PTEC of renal transplanted patients before and after conversion to PSI.

Methods To retrieve the maximal expression of our interest molecules in renal PTECs, we optimized antigen retrieval (AR) method and primary antibody dilution for each molecule separately. An optimization experiment was performed on 3 different normal patients renal biopsies were used. Later, human renal biopsy specimens originated from 4 different renal transplanted patients were used in this study. From all the 4 patients biopsy specimens were taken before and ongoing administration of PSIs (SRL, ERL). The expression of megalin, cubilin and albumin uptake in PTEC of renal transplant patients was determined by immunohistochemical staining.

Results Based on the optimization experiments, we selected the AR method and primary antibody dilution for the expression of megalin, cubilin and albumin uptake. In 4 renal transplanted patients following administration of PSIs results in patients 1, 2, 3 expression of megalin, cubilin and albumin uptake during ongoing PSI treatment was not comparable or even more intense than before PSIs introduction. The expression of megalin, cubilin and albumin uptake was reduced in patient 4 during ongoing PSI treatment.

Conclusion Our findings suggest that the renal transplant patient 4 developed proteinuria during PSI medication. The expression of megalin, cubilin and albumin uptake was markedly decreased during ongoing PSI treatment in patient 4. We concluded that there is a direct link between PSI medication and tubular dysfunction, which might cause proteinuria

The aim in this thesis is to use carbon, nitrogen and sulfur isotope analysis on teeth and bone to track diet and mobility of the population as well as the individual level from the articulated and disarticulated individuals from the passage grave Frälsegården, situated in Gökhem parish in the region of Falbygden, western Sweden. 50 human samples were analyzed from tooth, mandibula and femur, together with 11 faunal samples from different bone elements of local wild and domestic animals. The results fall well in line with earlier stable isotope work done on passage grave individuals from Falbygden in stable isotopes: A homogenous group with a diet which was clearly terrestrial and lived on a pretty high trophic level. It also seems that the analyzed individuals deliberately focused their food intake to the cultural and natural, megalith and limestone landscape of Falbygden, and not to its surroundings, even if the passage grave is situated in the outermost areas of the region. A small study compares the sulfur results to with earlier work done on strontium isotopes. The results show that the different methods might provide a deeper understanding in the interpretation of the methods, and therefore suggesting that further work should be done to compare the methods.

Cette thèse décrit les travaux effectués pour le développement du détecteur à scintillation du nanosatellite IGOSat. Sur la base des exigences du projet, un concept de détecteur a été validé, concept qui doit également être validé à la fois par la simulation et par l’expérience.IGOSat est un projet de satellite universitaire visant à développer un nanosatellite contenant une charge utile, basée sur un scintillateur, capable de mesurer le fond radiatif dans les zones aurorales et l’Anomalie de l’Atlantique Sud (ASA) en orbite terrestre basse. Le satellite dispose aussi d'une charge utile GPS bi-fréquence permettant de mesurer le contenu électronique total (TEC) de l'ionosphère. Ces deux charges utiles sont hébergées sur une plateforme 3U CubeSat qui sera lancée sur une orbite polaire à une altitude d’environ 650 km.La charge utile du scintillateur comprend un détecteur composé d'un cristal scintillant inorganique entouré de cinq scintillateurs organiques. Le scintillateur inorganique choisi est un cristal de CeBr3, sensible à la fois aux photons gamma et aux électrons. Les cinq scintillateurs plastiques EJ-200, qui sont principalement sensibles aux électrons, ont été choisis pour permettre de distinguer les deux types de particules. En d'autres termes, nous pouvons dire qu'une particule est un photon gamma lorsqu'elle n'interagit que dans le cristal de CeBr3, alors qu'il s'agit d'un électron si au moins une interaction est survenue dans un des scintillateurs plastique.Une simulation Monte-Carlo a été utilisée pour étudier la capacité de détection de ce détecteur. Une matrice de réponse a été élaborée pour la simulation des rayons gamma, qui peut être utilisée pour estimer le spectre en énergie des photons gamma en orbite terrestre basse.Un banc d'essai expérimental a été mis en place pour mesurer le spectre de rayonnement de sources radioactives. Ces mesures servent non seulement à valider le résultat de la simulation, mais également à en déduire la résolution en énergie du détecteur et une méthode d’étalonnage correspondante.Une comparaison entre les simulations de Monte-Carlo et les mesures expérimentales est également fournie dans cette thèse.Sur la base des travaux décrits ci-dessus, la thèse est constituée de 6 chapitres énumérés ci-dessous:- Le chapitre 1 est l'introduction au projet et une revue des études sur le fond radiatif en orbite terrestre basse, ainsi que les activités de développement CubeSat.- Le chapitre 2 décrit la configuration su satellite IGOSat, qui est développé pour remplir les exigences de fonctionnement des charges utiles.- Le chapitre 3 décrit le concept du détecteur à scintillation, son système électronique de lecture et le concept opérationnel de la charge utile.- Le chapitre 4 explique les processus physiques d’interaction d'une particule dans les matériaux à scintillation, la simulation de Monte-Carlo et la matrice de réponse du détecteur IGOSat.- Le chapitre 5 fournit les résultats des mesures expérimentales, issues des bancs de tests spécifiques mis en place. Les comparaisons entre simulation et expérience sont également décrites à la fin de ce chapitre.Le chapitre 6 est la conclusion de ce travail
This dissertation describes the work that has been done for the development of the scintillation detector of the IGOSat nanosatellite. Based on the requirements of the project, a concept of the detector has been proposed which required the validation by both simulations and experiments.IGOSat is a university satellite project aimed at developing a nanosatellite containing a scintillator payload that can measure the radiative background in the aurora zones and the South Atlantic Anomaly (SAA) on Low-Earth Orbit. Beside that, the satellite has a GPS Dual-Frequency Payload for measuring the Total Electronics Content (TEC) in the ionosphere. These two payloads are hosted on a 3U CubeSat platform that will be launched on a polar orbit at an altitude of about 650 km.The scintillator payload included a detector which is composed of an inorganic scintillation crystal and five surrounding organic scintillators. The chosen inorganic scintillator is Cerium Bromide (CeBr3), which is sensitive to both gamma-ray photons and electrons. The five surrounding EJ-200 plastic scintillators, which are mainly sensitive to the electrons, are chosen to discriminate the two types of particles. In other words we can say a particle is a gamma-ray photon when it interacts only in the CeBr3 crystal, while it is an electron if at least one interaction happened in a plastic scintillator.Monte-Carlo simulations have been used to investigate the detection ability of this detector. A response matrix has been made for the gamma-ray simulation, which can be used to estimate the original energy spectrum of the Low-Earth Orbit gamma-ray photons.An experimental test bench has been set up for measuring the detected spectrum of radioactive sources. These measurements are not only used to validate the simulation results, but also to determine the energy resolution of the detector, and a calibration method for it.A comparison between the Monte-Carlo simulations and the experimental measurements is also provided in this dissertation.Based on the topics described above, the dissertation has 6 chapters as listed below:- Chapter 1 is the introduction to the project and a review of the studies on Low-Earth Orbit radiative background, as well as description on the CubeSat development activities.- Chapter 2 describes the IGOSat satellite configuration on the platform, which is developed to support the payload based on their requirements.- Chapter 3 describes the concept of the scintillation detector, its electronics readout system and the operational concept of the payload.- Chapter 4 explains the physical processes of a particle in the scintillation materials, the Monte-Carlo simulations and the response matrix of the IGOSat detector.- Chapter 5 provides the experimental measurement results, based on each test bench that has been set up. The comparisons between simulation and experiment are also described at the end of this chapter.Chapter 6 is the conclusion of the work

The present thesis offers an innovative perspective analysing the perception of Turkish atrocities against the Ottoman Greek communities during the temporal period May 1919-December 1922 through the Athenian newspapers Estia and Empros, which were committed to the ubiquitous Greek irredentist vision of the Megali Idea. Delving into theories which emphasize on the political nature of nationalism, on national mobilisation and on the exploitation of mass communication by the elite, this thesis attempts to elaborate on the inclusion of the Ottoman Greeks into an expanded Greek nation-state, based on the principle of self-determination. Hence, from the Greek Press' perspective, the presented Turkish atrocities against a part of the Greek nation and potential subjects of a broadened Greek state had been contextualized on the basis of Greek nationalism. The study shows that, despite the widespread and divided Greek communities in the Ottoman Empire/Turkey, Estia and Empros were declaring that their common denominator had been their Greek self-determination and the fact that all these communities were viewing the Greek state and the Greek army as their guarantors for their safety. Adding to the agents of Hellenism also the Ecumenical Patriarchate, Estia and Empros were highlighting the crucial role of these agents towards the endangered Ottoman Greek communities. Thus, the presentation of the respective information in the newspapers was leaning on this theoretical schema.

Dissertação de mestrado em Bioquímica Aplicada
The incidence of neurodegenerative diseases is increasing today, so there is an increasing interest and need to better understand the mechanisms involved. Transthyretin (TTR) is a protein that has several physiological functions, being highly involved in the homeostasis of the nervous system. Changes in TTR expression levels are associated with several neurodegenerative pathologies. In addition, TTR plays a crucial role in the preservation and survival of neurons since its internalization through the action of megalin, activates several signalling mechanisms. Megalin has been extensively studied over the years; changes in the levels of this protein are also associated with neurodegenerative pathologies, thus functioning as a biomarker. Furthermore, TTR is regulated by heat shock factor 1 (HSF-1). HSF-1 acts in response to thermal shock, as well as to environmental stress (including ROS presence) and in pathological conditions such as protein misfolding, thus being associated with neuronal stress conditions. Thus, one of the aims of this work was to produce an antibody specific for soluble non membranar megalin and to characterize it so that one could quantify the levels of megalin in mice samples. The anti-megalin antibody was characterized by immunohistochemistry (IHC), by enzymelinked immunosorbent assays (ELISA) and Western blot; it was used to study megalin expression in the brain of wild-type (WT), TTR-knockout (TTR-KO) and HSF-1-deficient mice (with and without TTR), with and without an oxidative diet. The results show that megalin is expressed in the brain, especially in the retrosplenial area of the cortex. Additionally, WT animals present higher levels of megalin in this region than other stains, namely TTR-KOs and HSF-1-deficients. Furthermore, untreated animals presented higher levels of megalin expression in comparison to treated animals. The results suggest that i) the oxidative diet has an impact on the expression of megalin and ii) TTR influences the expression of megalin. The implication of megalin expression and its modulation in the retrosplenial area of the cortex deserves further investigation.
A incidência das doenças neurodegenerativas é cada vez mais elevada nos dias de hoje, pelo que cada vez é maior o interesse e a necessidade de se compreender melhor os mecanismos envolventes. A transtirretina (TTR) é uma proteína que apresenta diversas funções fisiológicas, estando altamente envolvida na homeostasia do sistema nervoso. Alterações nos níveis de expressão de TTR estão associadas a diversas patologias neurodegenerativas. Além disso, a TTR desempenha um papel crucial na preservação e sobrevivência dos neurônios uma vez que a sua internalização pela ação da megalina, ativa mecanismos de sinalização. A megalina tem sido um alvo de estudo uma vez que alterações nos níveis desta estão também associadas a patologias neurodegenerativas, funcionando por isso como um biomarcador. A TTR é também regulada pelo fator de transcrição de choque térmico 1 (HSF-1). HSF-1 para além de atuar na resposta ao choque térmico, atua como resposta ao stress (como na presença de espécies reativas de oxigénio) e em condições patológicas como no mau-enrolamento de proteínas, estando associado a condições de stress neuronal. Desta forma, um dos objetivos deste trabalho foi produzir um anticorpo específico para a megalina solúvel e caraterizá-lo para que com ele se pudesse analisar os níveis de megalina em amostras de ratinho. A caraterização do anticorpo foi feita por estudos de imunohistoquímica, por ensaios de imunoabsorção enzimática e Western blot. A expressão da megalina foi estuda em grupos de animais com TTR (WT), sem TTR (TTR-KO) e animais com ausência de HSF-1 (com e sem TTR), na presença e ausência de uma dieta oxidativa. Os resultados obtidos mostram que a megalina é expressa no cérebro nomeadamente na área retrosplenial do córtex. Além disso, verificou-se que os animais WT apresentam níveis mais altos de megalina nesta zona do córtex do que as outras estirpes, nomeadamente em animais com ausência de TTR e com falta de HSF-1 (quer com e sem TTR). Verificou-se também que os animais sujeitos à dieta apresentam menor expressão de megalina que os animais sem sujeitos à dieta. Os resultados sugerem que: i) a dieta oxidativa tem impacto na expressão de megalina e ii) a TTR influencia a expressão de megalina. Estudos sobre a interferência da expressão de megalina e a sua modulação na área retrosplenial do córtex devem ser desenvolvidos.

碩士
國立陽明大學
神經科學研究所
101
Statins, which are 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, reduce both cholesterol and isoprenoid synthesis and have been used to treat patients with diseases associated with hypercholesterolemia. Moreover, previous studies have demonstrated their therapeutical effectiveness in some, but not all, patients suffering Alzheimer’s disease. Previous studies from this laboratory have demonstrated that lovastain increases cellular levels of RhoA and serum-derived bovine serum albumin (BSA) in astrocytes, but not in neurons. This study thus aims to explore the mechanism(s) by which lovastatin modulates RhoA expression and albumin endocytosis in astrocytes. In this study, we found that lovastatin increases the cellular content of serum-derived bovine serum albumin (BSA) in cortical astrocytes. Immunoprecipitation and immunocytochemical studies reveal that lovastatin treatment increases the interaction of BSA with megalin, an endocytic receptor for multiple ligands including albumin. In addition, lovastatin increase the internalization of biotin-labeled surface proteins and the cellular content of BSA after BSA challenge. Interestingly, elevated level of RhoA was observed in astrocytes treated with lovastatin for 24 hr when the cellular BSA content was not affected. The effect of lovastatin on BSA, in fact, became apparent at a later time point, 48 hr. As noted, it was also found that lovastatin caused accumulation of ubiquitin-labeled proteins in a temporal profile similar to the increase in RhoA content. Moreover, mevalonate prevented lovastatin from increasing both RhoA and ubiquitin-labeled proteins. However, lovastatin didn’t decrease chymotrypsin-like proteasome activity. Together, results from this work demonstrate the involvement of ubiquitin and megalin in elevated levels of RhoA and internalized BSA, respectively, in astrocytes exposed to lovasatin.

碩士
國立成功大學
生物多樣性研究所
98
Many species of barbets are primarily frugivorous and potential or actual seed dispersers. Studies of food resources for barbets, nevertheless, remain little studied. The Taiwan barbet (Megalaima nuchalis Gould, 1863) is endemic to Taiwan and occurs in forest from lowlands to mid-elevation mountains. This study investigated food resource use by Taiwan barbets and its effects on seed dispersal. Direct observations were adopted to record foraging behaviors of barbets. Fruits of species that barbets fed on were also collected for morphometric measurements. I also conducted feeding and germination experiments to count seed retention time, and to estimate the effects of digestion, respectively. Taiwan barbets appeared to be generalists and obligate frugivores, and were opportunistic in searching fruits. Fruiting species with more number of fruiting trees and larger fruit biomass were visited more frequently by barbets, and the former factor was particularly important. Barbets revealed variable degrees of preferences for different fruiting species, with Myrica adenophora Hance as the most preferred species. In each visitation, barbets spent less than 5 min staying in fruiting trees, then left within 1 min after eating 1 - 3 fruits. The gape width of barbets set a limitation, roughly around 16 mm, under which barbets either swallowed or pecked a fruit, depending on its size. For 14 out of the 19 fruiting species visited by barbets, they took all ingested seeds away from materal trees, and up to 150 m. In germination experiments, no seeds of M. adenophora and Viburnum odoratissimum Ker germinated because of the mechanism of dormancy. Among other 17 fruit species, eleven (65 %) did not germinate, or showed only low germination percentage (＜ 10 %) when the seeds were in intact fruits, unless pulps were removed through artificial or digestion processes. The latter treatments improved both the success rate and speed of seed germination. Further analysis indicated the deinhibition effect as the major mechanism promoting seed germination, rather than the scarification effect. My results indicate that Taiwan Barbet can be considered as a good seed disperser, and is capable of improving seed germination. The distance of seeds actually dispersed from parent trees by barbets and its effects require further investigations.

Talayotic architecture arose on The Balearic Islands at the end of 2. millenium as syntesis of local megalitic culture of Bronze Age and cultural influence from central Mediterraen Sea, mainly from Sardinia and Corsica. The core of talayotic buildings was a naveta and megalithic tombs, whose using were abandoned just at the beginning of Iron Age. The talayotic architecture was developing on proper way in the manner of the insular cultures of occidental Mediterraen Sea until the Fenicians arrived ,in the same time the construction of talayots had declined. On the other hand the santuaries and taulas had reached their full using. Approximately in the 6 century in postalayotic period megalitic culture disapeared definitely. Last talayotic site were destroyed during Roman colonization.

Dissertação de mest., Estudos Marinhos e Costeiros, Faculdade de Ciências do Mar e do Ambiente, Univ. do Algarve, 2005
Este trabalho pretende descrever os padrões de distribuição horizontal, vertical e temporal dos estádios larvares de Pisidia longicornis e a sua relação com as condições hidrológicas na plataforma continental Portuguesa. Os dados aqui apresentados foram obtidos a partir de uma campanha oceanográfica realizada ao largo da Ria de Aveiro, entre os dias 9-22 de Maio de 2002. Na análise temporal foram também utilizados dados históricos referentes ao período de Outubro de 1986 a Janeiro de 1989. Os resultados permitem concluir que o estádio recém eclodido é transportado da linha de costa até à plataforma continental, conseguindo manter-se durante o seu desenvolvimento entre os 20-25 km da costa, através de migrações verticais favorecidas pela zona de retenção aí existente. As migrações verticais fazem-se pela subida na coluna de água no período nocturno, com ritmos diferentes consoante o estádio de desenvolvimento. O zoé I, sem um ritmo claro de migração vertical, ascende desde os 25 m de profundidade até à camada superficial. O zoé II inicia a sua migração logo após o pôr-do-sol, desde perto do fundo até aos 10-15 m. O megalopa migra verticalmente na coluna de água entre as 0 e as 4h, desde o fundo até à camada de neuston. Espacialmente, comparando com os estádios de zoé, o megalopa encontra-se em maior abundância nas estações mais próximas da costa, evidenciando um transporte efectivo para os locais de assentamento. A evolução mensal de abundâncias larvares ao longo do ano na área em estudo sugere dois picos de emissão larvar (Março- Abril, Julho-Setembro), reflectindo dois picos de abundância do megalopa (Primavera e final do Verão)

The term 'megalith' takes its origins from the ancient Greek 'megas' ('great') and 'lithos' ('stones'), and was first used by Algernon Herbert to describe Stonehenge. Yet since the term's inception, a multitude of megalithic forms have been identified across the world, the earliest dating to c. 12,000 BP at Gobekli Tepe in Turkey. Ranging from menhirs and trilithons to dolmens and cromlechs, these large stone structures have inspired fascination in those that have beheld them. The catalyst for this awe not simply being due to their immense scale, but also due to what they represent; namely past beliefs regarding life, death, and a person's place in the universe. Perhaps the most distinctive megalithic form is the jar, being both atypical and enigmatic. A quintessential example of this particular style can be found on the Plain of Jars, located in north-central Laos, though instances of similar jars have also been recorded in India, Myanmar, and Sulawesi. Groslier described these Laotian jars as "made of white sandstone...three to ten feet high, and often covered with a round lid. They served as tombs; [with] human ashes [being] placed inside". Spread across a sizeable area within Lao PDR, over 2000 megaliths have been recorded at 93 distinct locations, with further unconfirmed sites potentially leading to a total 118 sites. However, these large vessels have often been characterised as a mystery, with this uncertainty focusing on their origin, age, and overall function. Yet with the recent expansion of scientific investigation onto the Plain of Jars, a greater understanding of the sites has begun to develop. The aim of this dissertation is to analyse two major aspects of megalith significance, namely their design and placement. In this sense, megalithic 'design' encapsulates the morphology and scale of specimens studied within this project. This centres on the corresponding analysis of megalith stylisation, and the dimensions of those vessels. While simultaneously, the concept of specimen 'placement' considers the spatial positioning of megalithic material culture. Principally, this will discern whether the size and stylisation of jars and stone discs influenced their placement and arrangement. The primary means in achieving this is the application of the newly developed 'Type-Space model'. This paradigm focuses on the in-depth analysis of megalith morphology, and its superimposition onto high-resolution spatial data. Consequently, this duality provides an unparalleled understanding of the scale and decoration of the Plain of Jars megaliths, whilst also generating a unique image of their configuration.

The diploma thesis Celts and their influence on modern European cultures engages in Celtic nations, describes their history and presents remainders of Celtic culture in modern cultures. The thesis is focused on the influence of the Celts on the French region of Brittany and the Czech Republic. It mainly describes the heritage of the Celtic culture in the contemporary society in the form of Druidism, feasts, perception of nature and other traditions. It aims to create a complex image of the Celtic civilization. The second objective of the thesis is to confirm or refute the original connection of the modern Celtic cultural elements on a historical culture of the Celts. It achieves these aims on the basis of analysis, synthesis and subsequent comparison of obtained information about historical and contemporary Celts. The diploma thesis comes to the conclusion that all studied cultural elements have a real historical basis, but most of them, in their modern form, are created artificially. Nevertheless this fact does not detract from the importance of the cultural heritage of the Celts. KEYWORDS Celts, Brittany, the Czech Republic, megaliths, druids, feasts, traditions, cultural heritage of Celts

English, Zulu and Northern Sotho summaries
This study aims to evaluate the Non-Ferrous Metals Crime Combating Committee's impact on combating copper cable theft in South Africa. Data was collected via in-depth interviews with awaiting trial detainees charged with copper cable theft at the Pollsmoor Correctional Centre in the Western Cape; members of the Mpumalanga and Gauteng Provincial Non-Ferrous Metals Crime Combating Committee; members of Business Against Crime South Africa; and investigators attached to Combined Private Investigations who investigate copper theft nationally. The in-depth interviews provided a comprehensive understanding of participant experiences relating to the impact of the Non-Ferrous Metals Crime Combating Committee on the combating of copper cable theft in South Africa. Moreover, the researcher performed a thorough literature study of the phenomenon locally and internationally. Research findings indicate shortcomings in the Non-Ferrous Metals Crime Combating Committee's effectiveness, limiting its impact on preventing copper cable theft. Based on these findings, the study recommends that the Non-Ferrous Metals Crime Combating Committee be replaced with a specialised South African Police Service Non-Ferrous Metals Theft Unit. Consequently, this study puts forward a set of recommendations proposing a systematic pro-active plan to address and minimise copper cable theft in South Africa. The proposed plan convincingly presents practical solutions to minimise copper cable theft and contributes to the current body of scholarship on copper cable theft in South Africa.
Inhloso yalolu cwaningo ukuhlola umthelela wohlelo lwe-Non-Ferrous Metals Crime Combating Committee mayelana nokuqedwa kokutshontshwa kwentambo yogesi (copper cable). Idatha iqoqwe ngokusebenzisa izinhlolovo ezijulile ezenziwa ngokusebenzisa iziboshwa eziboshelwe izintambo zikagesi ezisamele ukugwetshwa ejele lase Pollsmoor Correctional Centre ngaseNtshonalanga Kapa, amalunga eMpumalanga and Gauteng Provincial Non-Ferrous Metals Crime Combating Committee, amalunga eBusiness Against Crime South Africa kanye nabaphenyi abahambisana nophiko lweCombined Private Investigations oluphenya ukutshontshwa kwezintambo zikagesi ezweni lonke. Lezi zinhlolovo ezijulile zinikeze ulwazi olubanzi lwabadlalindima olumayelana nomthelela weNon-Ferrous Metals Crime Combating Committee mayelana nokuqedwa kokutshontshwa kwezintambo zikagesi. Ngaphezu kwalokho, umcwaningi wenze ucwaningo olunzulu lombhalo wobuciko mayelana nokutshontshwa kwezintambo zikagesi lapha ekhaya kanye nasemhlabeni wonke jikelele. Ulwazi olutholakele lukhombisa iziqi ezikhinyabeza ukusebenza kahle kweKomithi mayelana nokuvimbela umthelela walo mayelana nokutshontshwa kwezintambo zikagesi. Ngezizathu ezisuselwa phezu kwalolu lwazi, lolu cwaningo lubeka phambili isethi lezincomo ezinganceda iKomiti ekuqiniseni umthintela walo ngokuhlongoza uhlelo olugqugquzelayo lokunciphisa izinga lokutshontshwa kwezintambo zikagesi eNingizimu Afrika. Lolu hlelo oluhlongoziwe ngaphandle kokungabaza lwethula izixazululo ezenzeka empilweni yangempela ukunciphisa ukwetshiwa kwezintambo zikagesi kanti lokhu kunegalelo kwiziko lamanje futhi luqhubekisa iziko lamanje lwezifundo esimayelana nokwetshiwa kwezintambo zikagesi.
Maikemisetso a thutelo ye ke go lekola khuetso ya Komiti ya Twantsho ya Bosenyi bja Dimetale tse di Sego tsa Tshipi ka ga go lwantsha kutso ya megala ya koporo. Datha e kgobokeditswe ka mokgwa wa ditherisano tse di tseneletsego tseo di sepeditswego le mahodu a megala ya koporo ao a golegilwego ao a letetsego go sekiswa Senthareng ya Tshokollo ya Pollsmoor go la Kapa Bodikela, maloko a Diprofense tsa Mpumalanga le Gauteng a Komiti ya Twantsho ya Bosenyi bja Dimetale tse di sego tsa Tshipi, maloko a Dikgwebo tsa Kgahlanong le Bosenyi tsa Afrika Borwa le banyakisisi bao ba dirisanago le Dinyakisiso tsa Praebete tse Kopantswego tseo di nyakisisago kutso ya koporo kemong ya bosetshaba. Ditherisano tse di tseneletsego tse di file kwesiso ka botlalo ya maitemogelo a batseakarolo ye e sepelelanago le khuetso ya Komiti ya Twantsho ya Bosenyi bja Dimetale tse di sego tsa Tshipi ka ga go lwantsha kutso ya megala ya koporo. Gape, monyakisisi o dirile thutelo ya dingwalo ka botebo ya kutso ya megala ya koporo tikologong ya leagong le kemong ya boditshabatshaba. Dikhwetso tsa dinyakisiso di supa mafokodi ao a lebanego bokgoni bja Komiti tshitisong ya khuetso ya yona go kutso ya megala ya koporo. Go ya ka dikhwetso tse, thutelo ye e hlagisa sehlopha sa dikeletso tseo di ka thusago Komiti go tswetsa khuetso ya yona pele ka go sisinya leano la go itokisetsa seemo se ka diregago ka mokgwa wo o beakantswego go fokotsa kutso ya megala ya koporo ka Afrika Borwa. Leano leo le sisintswego le hlagisa ka mo go kgodisago ditharollo tse di ka phethagatswago go fokotsa kutso ya megala ya koporo ka gona la ba la seabe go le go tswetsa pele popego ya bjale ya borutegi ka ga kutso ya megala ya koporo.
Criminology and Security Science
Ph. D. (Criminal Justice)