No description available.

Although the risk of malformations in the offspring of diabetic mothers remains the leading cause of perinatal mortality, the pathogenesis has not been elucidated. The hypothesis of this study was that protein glycation and oxygen free radicals play a role in hyperglycemia-induced malformations. CD-1 mouse embryos (0-2 somites) were cultured under hyperglycemic conditions for 48 hours with the exogenous addition of anti-glycating agents and oxygen free radical scavengers. The exogenous addition of aspirin (ASA) and D-lysine reduced significantly the malformations and embryonic glycated protein levels. Salicylate, arachidonic acid (AA), and to a lesser extent, indomethacin also exerted protective effects, but with no effect on glycated protein levels. We hypothesize that ASA, salicylate and indomethacin are protective by exerting free radical scavenging action; and ASA and D-lysine are acting as potent anti-glycating agents. Moreover, we suggest that AA may have inhibited hyperglycemia-induced malformations through the protective action of prostaglandins against free radical damage. Serum media lipid peroxidation (LPO) was reduced in the ASA and indomethacin groups possibly due to either a direct free radical scavenging action and/or the inhibitory effects of these agents on cyclooxygenase activity thereby decreasing the oxygen free radicals produced by this enzyme system. On the other hand, AA was associated with an increased level of LPO in the serum media. As the evidence has shown, the cause of hyperglycemia-induced malformations appears to be multifactorial and no one agent can completely eliminate the problem, although protective action can be exerted at different levels of the glycation-free radical cascade of tissue damage.

The chronic and severe executive functioning (EF) and self regulation deficits experienced by children with fetal alcohol spectrum disorders (FASD) are well documented and EF and self regulation have been identified as core targets for intervention. The goals of this dissertation were to: (i) examine the effects of a self regulation treatment for children with FASD on a range of EF measures (ii) examine neural markers of treatment outcome and determine if functional magnetic resonance imaging (fMRI) can differentiate treatment responders from nonresponders; (iii) determine if treatment effects generalize to child compliance and qualitatively examine the treatment process. Twenty-five children with FASD participated. Using a wait-list control design children were assigned to an immediate treatment (TXT; n = 12) or delayed treatment control (DTC; n = 13) condition. All children received an evaluation of EF and fMRI at baseline and 12-week follow-up. Parents also completed questionnaires assessing EF and behavior as well as a feedback questionnaire upon completion of treatment. A subset of parents tracked compliance over the course of their child’s therapy. For the TXT group only, parent questionnaires were readministered at 6 month follow-up. At 12-week follow-up, children in the TXT group displayed significant improvements in inhibitory control and social cognition. Additionally, parents reported improved behavioral and emotional regulation. This improvement, along with a further improvement in parent-rated inhibitory control, was maintained at the 6-month follow-up. Neuroplastic changes were also observed as the TXT group showed increased BOLD response in the right prefrontal cortex (PFC) and left caudate on a task of inhibitory control. When treatment responders were compared to nonresponders a pattern of increased BOLD response was found bilaterally in the PFC and left caudate. Compliance tracking revealed that self regulation therapy generalized to improved child compliance at home. Qualitative analysis indicated that perceived clinician competence, caregiver insight about child’s problems and caregiver perceptions of child’s insight about their problems, were the most commonly endorsed themes by caregivers. Results from this research signify that children with FASD are responsive to psychotherapy and following a brief intervention, showed improvements in self regulatory abilities that generalize to other EF areas and parent-reported behaviors.

Renal blood flow and micropuncture studies were conducted in late gestation fetal sheep (gestational age 134 - 141 days; term 150 days) and neonatal lambs (8 - 18 days after birth) to study the forces involved in glomerular filtration (GFR) and characterize the tubuloglomerular feedback (TGF) system during development. These studies required the kidney to be immobilized so stable models in acutely prepared anaesthetized animals were developed. Fetuses were studied in a heated water bath exteriorized from the uterus but with an intact umbilical circulation. The lower GFR in fetuses than lambs was found to be due to both lower net filtration pressures (P<0.001) and a lower ultrafiltration coefficient (P<0.001). TGF was present at both ages, but in fetuses the sensitivity was higher (P<0.001) and reactivity was lower (P<0.001). The reduction in TGF sensitivity between fetal and neonatal life may facilitate the increase in renal blood flow and GFR which occurs at this time. In both fetuses and lambs the sensitivity of the TGF curve was reduced by volume expansion (P<0.001, P<0.05) and reactivity was reduced in lambs (P<0.001). Furosemide abolished TGF at both ages. In both fetuses and lambs, TGF reactivity was increased by inhibition of neuronal nitric oxide synthase (nNOS; P<0.01, P<0.001) and in lambs, TGF sensitivity was increased (P<0.01). This indicates that nitric oxide produced by the macula densa modulates TGF during development. In offspring destined to become hypertensive due to maternal dexamethasone treatment in early gestation TGF sensitivity tended to be enhanced in fetal life and was enhanced in lambs (P<0.01). Increased TGF sensitivity may contribute to the development of hypertension in this model of developmental programming. The effects of nNOS inhibition were attenuated in these animals, suggesting that they have low tonic production of nitric oxide by the macula densa. In fetuses whose mothers had been subtotally nephrectomized prior to mating to induce maternal mild renal impairment, GFR was increased (P<0.01) but net filtration pressure was reduced (P<0.001) so the ultrafiltration coefficient was increased (P<0.001). TGF sensitivity was normal and the effects of nNOS inhibition were similar to normal fetuses.

Diss. (sammanfattning) Stockholm : Karol. inst., 2001. / Härtill 7 uppsatser.

Thesis (M.S.)--University of Wisconsin, 1983. / Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 85-90).

Thesis (Ph. D.)--University of Hawaii at Manoa, 2004. / Includes bibliographical references (leaves 92-105).

Thesis (M. Med. Sc.)--University of Hong Kong, 200.

Thesis (Ph.D.) - University of Queensland, 2006. / Includes bibliography.

This thesis describes investigations into Placental Protein 14 (PP14), a immunomodulator involved in human reproduction. The studies included the development of a purification procedure and an investigation of the activity of the protein. In addition the cDNA coding for the protein was cloned and expressed as a recombinant fusion protein and the molecular structure of the protein was predicted and analysed using computer-assisted modelling. Finally the clinical significance of the protein was studied in a range of patient groups. The purification scheme consisted of ion exchange, hydrophobic interaction and gel filtration chromatography, and the pure protein obtained was analysed by SDS-PAGE and Western blotting. The results demonstrate that the purification procedure is a suitable method to obtain PP14 in large quantity and with high purity. PP14 purified by this method retained its activity and was shown to suppress, in a dose-dependent manner, the uptake of 3H-Thymidine by peripheral blood mononuclear cells stimulated with interleukin-2. Purified PP14 was also shown to suppress the uptake of 3H-Thymidine by the cell line U937, also in a dose-dependent manner. This suppression could be removed by the incubation of the PP14 sample with an immunoabsorbent gel linked to monoclonal antibodies against PP14, demonstrating that PP14 was the molecule responsible for the observed activity. Based on the suppression by PP14 of U937 cell growth a bioassay for PP14 was developed, this assay was used to express the specific activity of PPM in Units/ml. To obtain recombinant PP14, mRNA was purified from a tissue sample and reverse transcription used to prepare cDNA. Specific primers were used to amplify the portion of cDNA coding for PP14 which was then ligated into the plasmids pUC 18 and pGEX-KG. Recombinant PP14 was then expressed as a fusion protein with glutathione-S-transferase. The expression conditions were optimised and the fusion protein was purified using affinity chromatography. The structure of PPM was investigated using computer assisted modelling. PPM is a member of the lipocalin family of proteins which share the feature of binding small hydrophobic molecules. The X-ray coordinates of two lipocalins known to share sequence homology with PP14 were used as a basis to model a predicted structure for PP14. An analysis of the structural motifs of the protein was carried out, and it was established that PP14 shares many of the characteristic features of this family of proteins including the presence of a binding pocket. The model was then used to predict potential ligands for PP14.PP14 was measured by radioimmunoassay in uterine flushings from fertile women, women with unexplained infertility and women suffering from recurrent miscarriages, and in plasma samples from fertile and infertile women. The results from the uterine flushings from fertile women showed that PP14 levels rose during the second half of the menstrual cycle reaching ug/ml levels by the end of the cycle. These physiological concentrations are in the same range as the concentrations at which the immunomodulatory activity of PP14 was observed in vitro. The levels of PP14 measured in uterine flushings were lower in infertile women than in fertile women, indicating that a deficiency in PP14 may be associated with infertility. The levels measured in plasma samples from these two groups of women did not pick up this difference. These results suggests that the measurement of proteins such as PP14 in uterine flushings instead of plasma samples may be a more sensitive indicator of local uterine function. In women suffering from recurrent miscarriage a significant lack of secretion of PP14 was observed around the time of implantation. This may be conected with the failure of implantation in these patients. A correlation was observed between the PP14 levels measured in uterine flushings from recurrent miscarriage patients and the level of endometrial development.