No description available.

Staigios kūdikio mirties sindromo diagnostika ir etiopatogenezė taikant šiuolaikinius metodus išlieka ribota ir net iki 0,5% mirties priežasčių nepavyksta nustatyti, atsakoma ne į visus klausimus susijusius su sindromo išsivystymu ir diagnoze. Viena iš šiuolaikinių metodų perspektyvų yra imunohistocheminių bei molekulinių metodų taikymas diagnozės patvirtinimui arba atmetimui. Minėti tyrimai yra labai svarbūs priežastinių ryšių nustatymui ir etiopatogenezės pagrindimui. Medžiagų apykaitos sutrikimai, tokie kaip glikogenozės I tipo liga – viena iš aktualiausių ir dažniausiai pasitaikančių, tačiau mažiausiai ištyrinėtų ligų. Darbo tikslas diagnozuoti glikogenozės I tipo atvejį taikant naujausias imunohistocheminių bei molekulinių tyrimų metodologijas. Tyrime naudojama naujausia parafine įlietų audinių technologija, audiniai įvertinti histologiškai, atliktos imunohistocheminės reakcijos. Iš parafine įlietų audinių išskirta DNR, kokybė įvertinta atlikus spektrofotometrinį tyrimą ir elektroforezę gelyje. Išskyrus DNR iš širdies ir kepenų buvo atlikta polimerazės grandininė reakcija naudojant tikslinius glikogenozės I tipo pradmenis. Gauti rezultatai parodė glikogeno kiekio padidėjimą kepenyse. Histologiniuose preparatuose matoma kasos, širdies miokardo, antinksčių hipertrofijos bei hiperplazijos. Imunohistocheminio tyrimo metu pastebėtas glikogeno kaupimasis kasos ląstelėse. Naudojantis gliukozės-6-fosfatazės žymenimis PGR metodu glikogenozės atvejo patvirtinti nepavyko. Iškeltos... [toliau žr. visą tekstą] / Diagnosis and etiopathogenesis of Sudden infant death syndrome are limited even nowadays, and up to 0.5 % of death the cause remains undiagnosed, not all the questions, related to the syndrome and diagnosis are answered. One of the applicable methods and perspectives for diagnosis rejection or approval are immunohistochemistry analysis and molecular methods. A very important determination of causal links and proof of etiopathogenesis are very important and possible only after the already mentioned tests. Metabolic disorders, especially glycogenosis type I disease - one of the sorest and the most common, but least-studied cases. The aim of this study is to diagnose glycogenosis type I using the latest immunohistochemistry and molecular methods. The latest technology of paraffin embedded tissue was used in the study, tissues were evaluated histologically, immunohistochemical reaction was carried-out. DNA was extracted from paraffin embedded tissues, quality was measured by spectrophotometric analysis and gel electrophoresis. Using the heart and liver - DNA polymerase chain reaction was performed using targeted glycogenosis type I primers. The results showed an increase of glycogen in the liver. Pancreas, heart, adrenal hypertrophy and hyperplasia were detected in histological samples. During immunohistochemical study an accumulation of glycogen in pancreas cells was observed. Using glucose-6-phosphatase PCR markers of glycogenosis could not be confirmed. Hypotheses for further... [to full text]

This thesis tests the hypothesis that increased neuronal cell death in SIDS infants is related to the ability of risk factors, such as prone sleeping, to expose infants to intermittent hypercapnic hypoxia (IHH). Based on the hypothesis that the NMDA system is linked to neuronal death, by way of excitotoxicity, correlations were also sought between cell death and changes in NMDA receptor (NR1) expression in brainstem nuclei controlling cardiorespiratory function. The first aim of this study was to verify that increased neuronal cell death occurs in SIDS infants. To verify a piglet model of SIDS risk factors, brainstem changes were examined in piglets exposed to IHH, and comparisons were made to changes seen in SIDS infants. The NMDA receptor was characterised in controls for both the human infant and the piglet groups. Comparisons of neuronal changes were made with SIDS infants, and piglets exposed to IHH. Non-radioactive in-situ hybridisation and immunohistochemistry were performed on formalin fixed and paraffin embedded brainstem tissue to identify markers of cell death (caspase-3, active caspase-3, and TUNEL), and to examine NR1 mRNA and protein expressions. Staining was quantified using computerised image analysis software. Eight nuclei from the brainstem medulla (caudal in piglets, and mid in infants), and two nuclei from the rostral pons (infants) were studied. The first dataset included human infants aged 1-6 months with a diagnosis of SIDS (n=15) or non-SIDS (n=10). The second dataset comprised developing piglets aged 13-14 days, with controls (n=6), against those exposed to IHH for 2 (n=6) or 4 (n=5) days. Increased neuronal cell death was not verified in the SIDS infants, but abnormalities in NR1 expression were present in selected nuclei of the medulla. Piglets exposed to IHH had increased neuronal cell death and changes in NR1 in selected nuclei of the medulla. There was also a positive correlation between increased cell death and high NR1 levels. Preliminary data showed that SIDS infants who usually slept prone had some differences in NR1 compared to those who did not usually sleep prone. From these findings, it was concluded that IHH may underlie the abnormalities in NMDA receptor expression that are present in the brainstem of SIDS infants. Although IHH can induce an increase in neuronal cell death, its significance in the aetiology of SIDS is not known. In piglets, IHH induced cell death correlated with high NMDA expression in some brainstem nuclei, supporting the hypothesis that excitotoxicity may be involved in the mechanism for cell death. Moreover, this thesis presents for the first time, �preliminary pathological proof� of an association between prone sleeping and abnormal NMDA receptor expression in SIDS infants.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Sudden Infant Death Syndrome (SIDS) is the sudden death of an infant younger than one year of age that remains unexplained after a complete investigation. For these infants, many different reasons have been hypothesized as to the cause of these deaths including: inherent vulnerability and improper hypoxic arousal. Studies done in other laboratories have shown that there seems to be a reduction in the levels of the neurotransmitter serotonin (5-HT) in the neurons of the raphe, extra-raphe, and ventral populations along with projection sites of these neurons. The huge implications of 5-HT in the control of respiration, prompted animal model studies to further investigate a potential connection between 5-HT and SIDS. 5-HT deficient mice were engineered by knocking out the Pet-1 transcription factor so that knockout mice only retained 30-40% of their brainstem 5-HT neurons. By comparing these 5-HT deficient Pet-1 knockout mice to wild-type mice, it was demonstrated that 5-HT deficient mice failed to autoresuscitate themselves after repeated bouts of hypoxia. Intriguingly, these mice only experienced an autoresuscitation deficit during a specific time period during development. To further evaluate the pathological development behind this behavior issue, in the current study we utilized mice that have modified Pet-1-Flpe driver, Egr2-Cre driver, along with a knock-in RC::FPSit allele to observe 5-HT development in the brainstem in a mature adult and across the critical period (postnatal days 8 and 13- P8 and P13). The transgenic mouse model Pet1-Krox20 gives us a way of exploring a specific subset of 5-HT neurons that rise from the developmental rhombomeres r3 and r5. The use of the knock-in RC::FPSit allele allows us to view the axonal projections of these specific 5-HT neurons by utilizing the presynaptic marker synaptophysin-GFP. This model (PKSit) will allow us to target 5-HT neurons that are implicated in respiration. We chose to compare two projection targets of the PKSit 5-HT neuron subtype through the vulnerable period of development and mature adult mouse: the Locus Coeruleus (LC) and the Nucleus Tractus Solitarius (NTS). In this study we tested the amount of colabeling between 5-HT and GFP in the LC and NTS at P8, P13, as well as the mature adult. We hypothesize that the LC undergoes significant serotonergic axon development and increases colocalization with GFP labeled axon projections between the ages of P8 and P13. We sliced mouse brains and ran immunofluorescence before taking confocal images. By utilizing ImageJ software to run colocalization analysis on the images obtained, we were able to quantify the amount of 5-HT labeled axon projections that are colocalized with GFP labeled axon projections. The parameters we used to quantify the amount of colocalization include the Pearson's Coefficient (PC), Mander's Coefficient (M1/M2), Cytofluorograms, Costes' Method, and van Steensel's Cross-Correlation Coefficient (CCF). We found that the LC shows significant changes with age in the colocalization of 5-HT with GFP while the NTS does not exhibit significant changes with age. The significant changes found in the LC 5-HT/GFP expression between the ages of P8 and P13 suggest one possible cause of failure of arousal. At P8, this lack of 5-HT colabeling with GFP projections suggests that there is some development occurring, which prevents the proper function of 5-HT. At P13, there is a significant increase in the colabeling of 5-HT with GFP, which indicates that the Pet1-Krox20 lineage is actively using 5-HT. The colocalization studies demonstrate that as the mouse ages, the amount of 5-HT labeling with GFP-synaptophysin in the NTS stays the same. The lack of overlap even in mature adult mice suggests that the expression of 5-HT in GFP labeled projections is not necessary. This colocalization study shows that there is an effect of age on the development of the serotonergic system in the LC, but no effect of age in the NTS. While this demonstrates that there is a critical period of development in relation to the LC, it is only one aspect of why mice pups failed to respond to repeated bouts of hypoxia.

No description available.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24201号 / 医博第4895号 / 新制||医||1061(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 西浦 博, 教授 森田 智視, 教授 松村 由美 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

Background: The death of a child has been found to have long term consequences for both individual and family functioning. This is particularly true for bereaved siblings who have been found to be at increased risk of developing mental health difficulties in later life. Literature on parental bereavement proposes that the parenting phenomenon, such as replacement child syndrome, subsequent child syndrome and the parenting paradox, which can emerge after the death of a child, may account for this. However, there is very little research on these labels of observed parenting phenomenon and, as a result, any hypothesis offered remains under elaborated. In addition, limited evidence suggests that, due to the sudden, unexpected and unexplained nature of the loss, SIDS parents are more likely to experience a greater degree of distress and adjustment difficulties than other perinatally bereaved populations. Given this, it could be hypothesised that SIDS parents may be likely to experience these parenting phenomena. Despite this, however, SIDS remains a neglected area of research. Aims: As a consequence of this research gap, the study aims to explore mothers’ experiences of parenting in their transition from being a parent unaffected by Sudden Infant Death Syndrome to a parent affected by Sudden Infant Death Syndrome. Methodology: Semi-structured interviews were conducted with seven mothers who had experienced an incident of Sudden Infant Death Syndrome. The interviews were then transcribed and analysed using Interpretative Phenomenological Analysis (IPA). Results: Five master themes emerged from the analysis: ‘Channelling the Parent Within’, a naturally developing and responsive parenting style that is facilitated by internal mechanisms, such as flexibility and confidence; ‘Parenting Outside of Yourself’, a parenting style that develops in the aftermath of a SIDS event, which is characterised by self doubt and a reliance on external mechanisms such as reassurance and restriction; ‘Restoration Through You’, the restorative effect of the subsequent and surviving children, which allows vindication and re-establishes happiness; ‘The Bitter Restoration’, a restoration that encompasses internal knowledge and external evidence of loss, including a disrupted family composition and a continued awareness of existential threat; ‘A Disruptive Appreciation’, the development of a greater appreciation for the subsequent and surviving children that impacts discipline and incorporates indulgence. These, along with the subthemes contributing to them, are presented as a narrative account. Conclusion: The results imply that mothers who have experienced a SIDS event shift into a permissive and anxious style of parenting which is characterised by safety behaviours. A model of parenting in the aftermath of SIDS has been proposed in order to explain the underlying cognitions and processes which drive this behaviour and the factors which serve to maintain it. By doing this it is hoped that, when working with bereaved parents and siblings, clinicians will be better positioned to frame parenting practices and intervene at a cognitive level.