Sex steroid feedback regulation of pituitary gonadotropins during early secondary oocyte growth in coho salmon (Oncorhynchus kisutch) – a potential target of endocrine disruption.

Thesis (Ph.D.)--University of Washington, 2016-06

Euphorbia tirucalli L.–Comprehensive Characterization of a Drought Tolerant Plant with a Potential as Biofuel Source

Of late, decrease in mineral oil supplies has stimulated research on use of biomass as an alternative energy source. Climate change has brought problems such as increased drought and erratic rains. This, together with a rise in land degeneration problems with concomitant loss in soil fertility has inspired the scientific world to look for alternative bio-energy species. Euphorbia tirucalli L., a tree with C3/CAM metabolism in leaves/stem, can be cultivated on marginal, arid land and could be a good alternative source of biofuel. We analyzed a broad variety of E. tirucalli plants collected from different countries for their genetic diversity using AFLP. Physiological responses to induced drought stress were determined in a number of genotypes by monitoring growth parameters and influence on photosynthesis. For future breeding of economically interesting genotypes, rubber content and biogas production were quantified. Cluster analysis shows that the studied genotypes are divided into two groups, African and mostly non-African genotypes. Different genotypes respond significantly different to various levels of water. Malate measurement indicates that there is induction of CAM in leaves following drought stress. Rubber content varies strongly between genotypes. An investigation of the biogas production capacities of six E. tirucalli genotypes reveals biogas yields higher than from rapeseed but lower than maize silage.

"Secretary in Fashion" and "Love-Letters": a study of socio-cultural and pragmatic-linguistic features in the late 17th century English language

Epistolary manuals are conspicuous historical documents for the pedagogy of letter writing; however, their actual usage as manuals by letter writers is unknown. "Secretary in Fashion" by Serre (1668), an epistolary manual, and "Love-Letters between a Nobleman and his Sister" (1684), an epistolary novel attributed to Behn, both give insights into epistolary conventions. Their inception and nature is interesting, considering their historical context. Despite the Restoration of Charles II, 17th century England was in a confused political state; as a result, texts regarding social convention or politics interested contemporary readers (the novel is inspired by a scandal of Lord Grey, an ardent Whig opposing Charles II). Past epistolary studies focus on 18th rather than 17th century manuals; the latter is typically used as supplementary information. Similarly, past epistolary fiction studies focus on 18th century texts; moreover, linguistic studies on Behn and the novel are deficient. Thus, this study addresses the research questions: 1) What are the socio-cultural and pragmaticlinguistic features represented in "Secretary in Fashion"? 2) What are the socio-cultural and pragmatic-linguistic features represented in "Love-Letters between a Nobleman and his Sister", and do any of these features correlate with the features represented in "Secretary in Fashion"? How far do the characteristic linguistic features of "Love-Letters between a Nobleman and his Sister" correlate with the practices recommended by the manual? Both texts were qualitatively analysed from an historical pragmatic perspective, which observes the potential effects of the socio-cultural and historical context. Also, as the texts concern shared discourses, comparisons were made with Gricean and Politeness Theory. The results show that the manual is a typical 17th century epistolary manual, aligning particularly with the "Academies of Complements", as it concerns the social conventions of the gentry. The novel mainly upheld instructions on form and matter; deviations occurred due to the amatory nature of some letters, and the narrative force affecting the style. Unfortunately, neither research question elucidates the actual usage of manuals. However, this study does show the epistolary practices of two writers, within specific contexts. It reveals that their 17th century English language use is affected by socialisation, in terms of social conventions concerning social rank, age, and gender; therefore, context varies language use. Also, their popularity reveals the interests of the 17th century society. Interest in epistolary-related texts, surely piques the interest of the modern reader as to why such epistolary-related texts were interesting.

Evolution of neodymium isotopic signature of seawater during the Late Cretaceous: Implications for intermediate and deep circulation

Neodymium isotopic compositions (εNd) have been largely used for the last fifty years as a tracer of past ocean circulation, and more intensively during the last decade to investigate ocean circulation during the Cretaceous period. Despite a growing set of data, circulation patterns still remain unclear during this period. In particular, the identification of the deep-water masses and their spatial extension within the different oceanic basins are poorly constrained. In this study we present new deep-water εNd data inferred from the Nd isotope composition of fish remains and Fe-Mn oxyhydroxide coatings on foraminifera tests, along with new εNd data of residual (partly detrital) fraction recovered from DSDP sites 152 (Nicaraguan Rise), 258 (Naturaliste Plateau), 323 (Bellinghausen Abyssal Plain), and ODP sites 690 (Maud Rise) and 700 (East Georgia Basin, South Atlantic). The presence of abundant authigenic minerals in the sediments at sites 152 and 690 detected by XRD analyses may explain both middle rare earth element enrichments in the spectra of the residual fraction and the evolution of residual fraction εNd that mirror that of the bottom waters at the two sites. The results point towards a close correspondence between the bottom water εNd values of sites 258 and 700 from the late Turonian to the Santonian. Since the deep-water Nd isotope values at these two sites are also similar to those at other proto-Indian sites, we propose the existence of a common intermediate to deep-water water mass as early as the mid-Cretaceous. The water mass would have extended from the central part of the South Atlantic to the eastern part of proto-Indian ocean sites, beyond the Kerguelen Plateau. Furthermore, data from south and north of the Rio Grande Rise-Walvis Ridge complex (sites 700 and 530) are indistinguishable from the Turonian to Campanian, suggesting a common water mass since the Turonian at least. This view is supported by a reconstruction of the Rio Grande Rise-Walvis Ridge complex during the Turonian, highlighting the likely existence of a deep breach between the Rio Grande Rise and the proto-Walvis Ridge at that time. Thus deep-water circulation may have been possible between the different austral basins as early as the Turonian, despite the presence of potential oceanic barriers. Comparison of new seawater and residue εNd data on Nicaraguan Rise suggest a westward circulation of intermediate waters through the Caribbean Seaway during the Maastrichtian and Paleocene from the North Atlantic to the Pacific. This westward circulation reduced the Pacific water influence in the Atlantic, and was likely responsible for more uniform, less radiogenic εNd values in the North Atlantic after 80 Ma. Additionally, our data document an increasing trend observed in several oceanic basins during the Maastrichtian and the Paleocene, which is more pronounced in the North Pacific. Although the origin of this increase still remains unclear, it might be explained by an increase in the contribution of radiogenic material to upper ocean waters in the northern Pacific. By sinking to depth, these waters may have redistributed to some extent more radiogenic signatures to other ocean basins through deep-water exchanges.

Nutrients limiting the Algal Growth Potential (AGP) in the Gulf of Riga, eastern Baltic Sea, in spring and early summer 1996

In May, June and July 1996, samples wcre collected along one transect greatly influenced by river discharge (eastern side of the gulf), along one transect slightly influence by river discharge (western side), at one station Iocated in the mouth of the main river (River Daugava), at one station located in the center of the Gulf and at several nearshore locations of the western side. Ratios of rnolecular concentrations of in situ dissolved ioorganic nitrogen, phosphorus and silicon, as weIl as enrichment bioassays were llsed to dctcrrnine which nutrient (s) lirnited the potential biomass of phytoplankton. Both comparison of (NO.d-N02+NJ.L): P04 (DIN: DIP) values with Redfic1d's ratio and bioassay inspection led to the sarne conclusions. Phosphorus was clearly the nutrient most limiting for the potcntial biornass of test species in nitrogen- rich waters, which occurred in mid spring, in the upper layer of the southern-eastern part of the Gulf which is greatly influenced by river discharge. In late spring, with the decrease of the total DIN reserve, nitrogen and phosphorus showed an equallimiting role. In deeper layers of this area and out of the river plume (western side and central part of the gulf), nitrogen was the limiting nutrient. In summer, whcn river discharge was the lowest, a11 DIN concentrations but one ranged between 1.6 and 2.6 µM, and the whole area was nitrogen-limited for both the cyanobacterial and the algal test strains. In 74% of the samples for which nitrogen was the limiting nutrient, phosphorus was recorded to be the second potentially limiting nutrient. In contrast, silicon never appeared as limiting the growth potential of either Microcystis aeruginosa or Phaeodactylum tricornutum; phosphorus was the limiting nutrient when DIN: Si03 values were >1 (in May), but DIN: Si03 was <1 when nitrogen was limiting (June and July). The authors conclude that the recently reported decrease of silicon loading in coastal waters and its subsequent enhanced importance in pushing the outcome of species competition towards harmful species may not yet be the most important factor for the Gulf of Riga. Iron appeared for 12% of the tests in the list of nutrients limiting the potential biomass. Tentative results also indicated that a significant fraction of the nitrogen (~,4 µg-atom N 1(-1) taken up by Microcystis aeruginosa may have been in the form of dissolved organic nitrogen (DON). It is thus also suggested tentatively that more attention be paid to these nitrients during further research in the Gulf of Riga.

Early and late effects of Ibuprofen on mouse sperm parameters, chromatin condensation, and DNA integrity in mice

Background: There are few studies indicating the detrimental effects of ibuprofen on sperm fertility potential and DNA integrity. Objective: To determine the effects of Ibuprofen on sperm parameters, chromatin condensation and DNA integrity of mice. Materials and Methods: In this experimental study, 36 adult male mice with average weight 37 gr were divided into three groups, including control (group I, n=12), normal dosage of ibuprofen (group II, n=12) and high dosage (group III, n=12). Ibuprofen with different doses was dissolved in daily water of animals. After 35, 70 and 105 days, the cauda epididymis of mice were cut and incubated in Ham’s F10 media. Sperm samples were analyzed for parameters (motility, morphology and count), DNA integrity (SCD test) and chromatin condensation (chromomycin A3 and Aniline blue staining). Results: After 35 days, in addition to above mentioned sperm parameters, all of the treated mice showed statistically significant increase in spermatozoa with immature chromatin (P<0.05). However, after 70 days, the rate of sperm DNA fragmentation assessed by SCD was increased in group II (66.5±0.7) and the percentage of immature spermatozoa (AB+ and CMA3+) was higher in group III (77.5±0.7 and 49.5±6.3 respectively) than other groups. After 105 days, the AB+ spermatozoa were increased in both normal dose and high dose groups. Conclusion: Ibuprofen may cause a significant reduction in sperm parameters and sperm chromatin/DNA integrity in mice. It should be noted that these deleterious effects are dose-dependent and can be seen in early and late stage of drug treatments.

Leaf water potential and sap flow as indicators of water stress in Crimson ‘seedless’ grapevines under different irrigation strategies

Vitis vinifera L. cv. Crimson Seedless is a late season red table grape developed in 1989, with a high market value and increasingly cultivated under protected environments to extend the availability of seedless table grapes into the late fall. The purpose of this work was to evaluate leaf water potential and sap flow as indicators of water stress in Crimson Seedless vines under standard and reduced irrigation strategy, consisting of 70 % of the standard irrigation depth. Additionally, two sub-treatments were applied, consisting of normal irrigation throughout the growing season and a short irrigation induced stress period between veraison and harvest. Leaf water potential measurements coherently signaled crop-available water variations caused by different irrigation treatments, suggesting that this plant-based method can be reliably used to identify water-stress conditions. The use of sap flow density data to establish a ratio based on a reference ‘well irrigated vine’ and less irrigated vines can potentially be used to signal differences in the transpiration rates, which may be suitable for improving irrigation management strategies while preventing undesirable levels of water stress. Although all four irrigation strategies resulted in the production of quality table grapes, significant differences (p ≤ 0.05) were found in both berry weight and sugar content between the standard irrigation and reduced irrigation treatments. Reduced irrigation increased slightly the average berry size as well as sugar content and technical maturity index. The 2-week irrigation stress period had a negative effect on these parameters.

Leaf water potential and sap flow as indicators of water stress in Crimson 'seedless' grapevines under different irrigation strategies

Abstract Vitis vinifera L. cv. Crimson Seedless is a late season red table grape developed in 1989, with a high market value and increasingly cultivated under protected environments to extend the availability of seedless table grapes into the late fall. The purpose of this work was to evaluate leaf water potential and sap flow as indicators of water stress in Crimson Seedless vines under standard and reduced irrigation strategy, consisting of 70 % of the standard irrigation depth. Additionally, two sub-treatments were applied, consisting of normal irrigation throughout the growing season and a short irrigation induced stress period between veraison and harvest. Leaf water potential measurements coherently signaled crop-available water variations caused by different irrigation treatments, suggesting that this plant-based method can be reliably used to identify water-stress conditions. The use of sap flow density data to establish a ratio based on a reference ‘well irrigated vine’ and less irrigated vines can potentially be used to signal differences in the transpiration rates, which may be suitable for improving irrigation management strategies while preventing undesirable levels of water stress. Although all four irrigation strategies resulted in the production of quality table grapes, significant differences (p ≤ 0.05) were found in both berry weight and sugar content between the standard irrigation and reduced irrigation treatments. Reduced irrigation increased slightly the average berry size as well as sugar content and technical maturity index. The 2-week irrigation stress period had a negative effect on these parameters.

Ecological responses of Western Palearctic avian species to late Quaternary climate oscillations

This PhD thesis explores the ecological responses of bird species to glacial-interglacial transitions during the late Quaternary in the Western Palearctic, using multiple approaches and at different scales, enhancing the importance of the bird fossil record and quantitative methods to elucidate biotic trends in relation to long-term climate changes. The taxonomic and taphonomic analyses of the avian fossil assemblages from four Italian Middle and Upper Pleistocene sedimentary successions (Grotta del Cavallo, Grotta di Fumane, Grotta di Castelcivita, and Grotta di Uluzzo C) allowed us to reconstruct local-scale patterns in birds’ response to climate changes. These bird assemblages are characterized by the presence of temperate species and by the occasional presence of cold-dwelling species during glacials, related to range shifts. These local patterns are supported by those identified at the continental scale. In this respect, I mapped the present-day and LGM climatic envelopes of species with different climatic requirements. The results show a substantial stability in the range of temperate species and pronounced changes in the range of cold-dwelling species, supported by their fossil records. Therefore, the responses to climate oscillations are highly related to the thermal niches of investigated species. I also clarified the dynamics of the presence of boreal and arctic bird species in Mediterranean Europe, due to southern range shifts, during the glacial phases. After a reassessment of the reliability of the existing fossil evidence, I show that this phenomenon is not as common as previously thought, with important implications for the paleoclimatic and paleoenvironmental significance of the targeted species. I have also been able to explore the potential of multivariate and rarefaction methods in the analyses of avian fossils from Grotta del Cavallo. These approaches helped to delineate the main drivers of taphonomic damages and the dynamics of species diversity in relation to climate-driven paleoenvironmental changes.

Late onset spinal cord ischemia after thoracoabdominal aortic aneurysm open repair

The aim of this study is to evaluate if spinal cord ischemia (SCI), especially its late presentation, and can be correlated to the results of intraoperative evoked potential monitoring (IOM). Methods. This study is a physician-initiated, retrospective, single-center, non-randomized study. Data from all patients undergoing a thoracoabdominal aortic aneurysm surgical repair (TAAA SR) between January 2016 and March 2020 IOM was collected and analyzed. Results. During the study period, 261 patients underwent TAAA SR with MEP/SSEPs monitoring [190 males, 73%; median age 65 (57-71)]. Thirty-seven patients suffered from SCI, for an overall rate of 14% (permanent 9%). When stratifying patients according to the SCI onset, 18 patients presented with an early (11 permanent) and 19 with a late SCI (<24h) (11 permanent). Of 261 patients undergoing TAAA SR with IOM, 15 were excluded due to changes in the upper extremity motor evoked potentials. For the remaining 246, the association between SCI and IOM was investigated: only irreversible IOM loss without peripheral changes have been found to be a risk factor for late onset SCI (p=.006). Furthermore, given that no statistical differences were found between the two groups when no IOM changes were recorded (p=.679), this situation cannot reliably rule out any SCI in our cohort. Independent risk factors for late spinal cord ischemia onset found at multivariate analysis were smoking history (p=.008), BMI>28 (p=.048) and TAAA extent II (p=.009). The irreversible MEP change without peripheral showed a trend of significance (p=.052). Conclusions. Evoked potential intraoperative monitoring is an important adjunct during thoracoabdominal aortic open repair to predict and possibly prevent spinal cord ischemia. Irreversible IOM loss without peripheral changes was predictive of late SCI, therefore more attention should be paid to the postoperative management of this subgroup of patients.

Investigating the causes of late fruit drop in ‘Regina’ sweet cherry (Prunus avium)

The presented study aimed to correctly describe the (late) fruit drop pattern of sweet cherry cv. Regina grafted on ‘Gisela 5’ and investigate its internal causes. In the first season, a method to describe the fruit drop pattern was defined and validated. The second season was devoted to a province-based screening of the phenomenon to identify potential influences of environmental, physiological and management factors. The multisite trial involved 6 commercial orchards located at different elevations, from 225 up to 1175m a.s.l. The third season was dedicated to find confirmation of the hypothesis formulated during the previous year. The multisite comparison was maintained but reduced to only two orchards to allow more frequent samplings. It emerged that late fruit drop is a complex phenomenon showing variable intensity: the percentage of late fruit drop ranged from 7 to 76% of the fruitlets set, depending on the orchard and on the year considered. Two main waves of fruitlets drop have been observed: the first one was composed by unfertilized parthenocarpic fruitlets, probably caused by late or missing fertilization, that immediately after bloom already showed smaller diameters and symptoms of senescence; the second one (the focus of this study) was composed by fully developed fruits that at a certain point decreased their growth rate and got senescent. All the late dropped cherries showed an aborted embryo. This sudden change has been observed to be concomitant both with prolonged periods of low temperatures (or sudden severe decreases in the daily Growing Degree Hours accumulation) and with extraordinary high temperatures close to or above 30°C. Other factors, such as the position of the limb within the canopy, its orientation (sunny vs. shady side) or nutrition played only a marginal role. Excessive vigor can increase late fruit drop intensity but is not its main cause.

Design, Synthesis And Biological Evaluation Of Hybrid Bioisoster Derivatives Of N-acylhydrazone And Furoxan Groups With Potential And Selective Anti-trypanosoma Cruzi Activity.

Hybrid bioisoster derivatives from N-acylhydrazones and furoxan groups were designed with the objective of obtaining at least a dual mechanism of action: cruzain inhibition and nitric oxide (NO) releasing activity. Fifteen designed compounds were synthesized varying the substitution in N-acylhydrazone and in furoxan group as well. They had its anti-Trypanosoma cruzi activity in amastigotes forms, NO releasing potential and inhibitory cruzain activity evaluated. The two most active compounds (6, 14) both in the parasite amastigotes and in the enzyme contain the nitro group in para position of the aromatic ring. The permeability screening in Caco-2 cell and cytotoxicity assay in human cells were performed for those most active compounds and both showed to be less cytotoxic than the reference drug, benznidazole. Compound 6 was the most promising, since besides activity it showed good permeability and selectivity index, higher than the reference drug. Thereby the compound 6 was considered as a possible candidate for additional studies.

Growth And Body Composition In Brazilian Female Rhythmic Gymnastics Athletes.

The aim was to analyse the physical growth and body composition of rhythmic gymnastics athletes relative to their level of somatic maturation. This was a cross-sectional study of 136 athletes on 23 teams from Brazil. Mass, standing height and sitting height were measured. Fat-free and fat masses, body fat percentages and ages of the predicted peak height velocity (PHV) were calculated. The z scores for mass were negative during all ages according to both WHO and Brazilian references, and that for standing height were also negative for all ages according to WHO reference but only until 12 years old according to Brazilian reference. The mean age of the predicted PHV was 12.1 years. The mean mass, standing and sitting heights, body fat percentage, fat-free mass and fat mass increased significantly until 4 to 5 years after the age of the PHV. Menarche was reached in only 26% of these athletes and mean age was 13.2 years. The mass was below the national reference standards, and the standing height was below only for the international reference, but they also had late recovery of mass and standing height during puberty. In conclusion, these athletes had a potential to gain mass and standing height several years after PHV, indicating late maturation.

Neonatal Outcomes Of Late Preterm And Early Term Birth.

To compare neonatal deaths and complications in infants born at 34-36 weeks and six days (late preterm: LPT) with those born at term (37-41 weeks and six days); to compare deaths of early term (37-38 weeks) versus late term (39-41 weeks and six days) infants; to search for any temporal trend in LPT rate. A retrospective cohort study of live births was conducted in the Campinas State University, Brazil, from January 2004 to December 2010. Multiple pregnancies, malformations and congenital diseases were excluded. Control for confounders was performed. The level of significance was set at p<0.05. After exclusions, there were 17,988 births (1653 late preterm and 16,345 term infants). A higher mortality in LPT versus term was observed, with an adjusted odds ratio (OR) of 5.29 (p<0.0001). Most complications were significantly associated with LPT births. There was a significant increase in LPT rate throughout the study period, but no significant trend in the rate of medically indicated deliveries. A higher mortality was observed in early term versus late term infants, with adjusted OR: 2.43 (p=0.038). LPT and early term infants have a significantly higher risk of death.

Antimicrobial And Antiproliferative Potential Of Anadenanthera Colubrina (vell.) Brenan.

The aim of the present study was to perform an in vitro analysis of the antimicrobial and antiproliferative potential of an extract from Anadenanthera colubrina (Vell.) Brenan (angico) and chemically characterize the crude extract. Antimicrobial action was evaluated based on the minimum inhibitory concentration (MIC), minimum bactericidal/fungicidal concentration, and the inhibition of formation to oral biofilm. Cell morphology was determined through scanning electron microscopy (SEM). Six strains of tumor cells were used for the determination of antiproliferative potential. The extract demonstrated strong antifungal activity against Candida albicans ATCC 18804 (MIC = 0.031 mg/mL), with similar activity found regarding the ethyl acetate fraction. The extract and active fraction also demonstrated the capacity to inhibit the formation of Candida albicans to oral biofilm after 48 hours, with median values equal to or greater than the control group, but the difference did not achieve statistical significance (P > 0.05). SEM revealed alterations in the cell morphology of the yeast. Regarding antiproliferative activity, the extract demonstrated cytostatic potential in all strains tested. The present findings suggest strong antifungal potential for Anadenanthera colubrina (Vell.) Brenan as well as a tendency toward diminishing the growth of human tumor cells.